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Sample records for calibrated automated thrombin

  1. Reproducibility, stability, and biological variability of thrombin generation using calibrated automated thrombography in healthy dogs.

    Science.gov (United States)

    Cuq, Benoît; Blois, Shauna L; Wood, R Darren; Monteith, Gabrielle; Abrams-Ogg, Anthony C; Bédard, Christian; Wood, Geoffrey A

    2018-06-01

    Thrombin plays a central role in hemostasis and thrombosis. Calibrated automated thrombography (CAT), a thrombin generation assay, may be a useful test for hemostatic disorders in dogs. To describe CAT results in a group of healthy dogs, and assess preanalytical variables and biological variability. Forty healthy dogs were enrolled. Lag time (Lag), time to peak (ttpeak), peak thrombin generation (peak), and endogenous thrombin potential (ETP) were measured. Direct jugular venipuncture and winged-needle catheter-assisted saphenous venipuncture were used to collect samples from each dog, and results were compared between methods. Sample stability at -80°C was assessed over 12 months in a subset of samples. Biological variability of CAT was assessed via nested ANOVA using samples obtained weekly from a subset of 9 dogs for 4 consecutive weeks. Samples for CAT were stable at -80°C over 12 months of storage. Samples collected via winged-needle catheter venipuncture showed poor repeatability compared to direct venipuncture samples; there was also poor agreement between the 2 sampling methods. Intra-individual variability of CAT parameters was below 25%; inter-individual variability ranged from 36.9% to 78.5%. Measurement of thrombin generation using CAT appears to be repeatable in healthy dogs, and samples are stable for at least 12 months when stored at -80°C. Direct venipuncture sampling is recommended for CAT. Low indices of individuality suggest that subject-based reference intervals are more suitable when interpreting CAT results. © 2018 American Society for Veterinary Clinical Pathology.

  2. Analytical and between-subject variation of thrombin generation measured by calibrated automated thrombography on plasma samples.

    Science.gov (United States)

    Kristensen, Anne F; Kristensen, Søren R; Falkmer, Ursula; Münster, Anna-Marie B; Pedersen, Shona

    2018-05-01

    The Calibrated Automated Thrombography (CAT) is an in vitro thrombin generation (TG) assay that holds promise as a valuable tool within clinical diagnostics. However, the technique has a considerable analytical variation, and we therefore, investigated the analytical and between-subject variation of CAT systematically. Moreover, we assess the application of an internal standard for normalization to diminish variation. 20 healthy volunteers donated one blood sample which was subsequently centrifuged, aliquoted and stored at -80 °C prior to analysis. The analytical variation was determined on eight runs, where plasma from the same seven volunteers was processed in triplicates, and for the between-subject variation, TG analysis was performed on plasma from all 20 volunteers. The trigger reagents used for the TG assays included both PPP reagent containing 5 pM tissue factor (TF) and PPPlow with 1 pM TF. Plasma, drawn from a single donor, was applied to all plates as an internal standard for each TG analysis, which subsequently was used for normalization. The total analytical variation for TG analysis performed with PPPlow reagent is 3-14% and 9-13% for PPP reagent. This variation can be minimally reduced by using an internal standard but mainly for ETP (endogenous thrombin potential). The between-subject variation is higher when using PPPlow than PPP and this variation is considerable higher than the analytical variation. TG has a rather high inherent analytical variation but considerable lower than the between-subject variation when using PPPlow as reagent.

  3. Automated Attitude Sensor Calibration: Progress and Plans

    Science.gov (United States)

    Sedlak, Joseph; Hashmall, Joseph

    2004-01-01

    This paper describes ongoing work a NASA/Goddard Space Flight Center to improve the quality of spacecraft attitude sensor calibration and reduce costs by automating parts of the calibration process. The new calibration software can autonomously preview data quality over a given time span, select a subset of the data for processing, perform the requested calibration, and output a report. This level of automation is currently being implemented for two specific applications: inertial reference unit (IRU) calibration and sensor alignment calibration. The IRU calibration utility makes use of a sequential version of the Davenport algorithm. This utility has been successfully tested with simulated and actual flight data. The alignment calibration is still in the early testing stage. Both utilities will be incorporated into the institutional attitude ground support system.

  4. Automated Calibration of Dosimeters for Diagnostic Radiology

    International Nuclear Information System (INIS)

    Romero Acosta, A.; Gutierrez Lores, S.

    2015-01-01

    Calibration of dosimeters for diagnostic radiology includes current and charge measurements, which are often repetitive. However, these measurements are usually done using modern electrometers, which are equipped with an RS-232 interface that enables instrument control from a computer. This paper presents an automated system aimed to the measurements for the calibration of dosimeters used in diagnostic radiology. A software application was developed, in order to achieve the acquisition of the electric charge readings, measured values of the monitor chamber, calculation of the calibration coefficient and issue of a calibration certificate. A primary data record file is filled and stored in the computer hard disk. The calibration method used was calibration by substitution. With this system, a better control over the calibration process is achieved and the need for human intervention is reduced. the automated system will be used in the calibration of dosimeters for diagnostic radiology at the Cuban Secondary Standard Dosimetry Laboratory of the Center for Radiation Protection and Hygiene. (Author)

  5. Increased Automation in Stereo Camera Calibration Techniques

    Directory of Open Access Journals (Sweden)

    Brandi House

    2006-08-01

    Full Text Available Robotic vision has become a very popular field in recent years due to the numerous promising applications it may enhance. However, errors within the cameras and in their perception of their environment can cause applications in robotics to fail. To help correct these internal and external imperfections, stereo camera calibrations are performed. There are currently many accurate methods of camera calibration available; however, most or all of them are time consuming and labor intensive. This research seeks to automate the most labor intensive aspects of a popular calibration technique developed by Jean-Yves Bouguet. His process requires manual selection of the extreme corners of a checkerboard pattern. The modified process uses embedded LEDs in the checkerboard pattern to act as active fiducials. Images are captured of the checkerboard with the LEDs on and off in rapid succession. The difference of the two images automatically highlights the location of the four extreme corners, and these corner locations take the place of the manual selections. With this modification to the calibration routine, upwards of eighty mouse clicks are eliminated per stereo calibration. Preliminary test results indicate that accuracy is not substantially affected by the modified procedure. Improved automation to camera calibration procedures may finally penetrate the barriers to the use of calibration in practice.

  6. System for Automated Calibration of Vector Modulators

    Science.gov (United States)

    Lux, James; Boas, Amy; Li, Samuel

    2009-01-01

    Vector modulators are used to impose baseband modulation on RF signals, but non-ideal behavior limits the overall performance. The non-ideal behavior of the vector modulator is compensated using data collected with the use of an automated test system driven by a LabVIEW program that systematically applies thousands of control-signal values to the device under test and collects RF measurement data. The technology innovation automates several steps in the process. First, an automated test system, using computer controlled digital-to-analog converters (DACs) and a computer-controlled vector network analyzer (VNA) systematically can apply different I and Q signals (which represent the complex number by which the RF signal is multiplied) to the vector modulator under test (VMUT), while measuring the RF performance specifically, gain and phase. The automated test system uses the LabVIEW software to control the test equipment, collect the data, and write it to a file. The input to the Lab - VIEW program is either user-input for systematic variation, or is provided in a file containing specific test values that should be fed to the VMUT. The output file contains both the control signals and the measured data. The second step is to post-process the file to determine the correction functions as needed. The result of the entire process is a tabular representation, which allows translation of a desired I/Q value to the required analog control signals to produce a particular RF behavior. In some applications, corrected performance is needed only for a limited range. If the vector modulator is being used as a phase shifter, there is only a need to correct I and Q values that represent points on a circle, not the entire plane. This innovation has been used to calibrate 2-GHz MMIC (monolithic microwave integrated circuit) vector modulators in the High EIRP Cluster Array project (EIRP is high effective isotropic radiated power). These calibrations were then used to create

  7. Individual differences in the calibration of trust in automation.

    Science.gov (United States)

    Pop, Vlad L; Shrewsbury, Alex; Durso, Francis T

    2015-06-01

    The objective was to determine whether operators with an expectancy that automation is trustworthy are better at calibrating their trust to changes in the capabilities of automation, and if so, why. Studies suggest that individual differences in automation expectancy may be able to account for why changes in the capabilities of automation lead to a substantial change in trust for some, yet only a small change for others. In a baggage screening task, 225 participants searched for weapons in 200 X-ray images of luggage. Participants were assisted by an automated decision aid exhibiting different levels of reliability. Measures of expectancy that automation is trustworthy were used in conjunction with subjective measures of trust and perceived reliability to identify individual differences in trust calibration. Operators with high expectancy that automation is trustworthy were more sensitive to changes (both increases and decreases) in automation reliability. This difference was eliminated by manipulating the causal attribution of automation errors. Attributing the cause of automation errors to factors external to the automation fosters an understanding of tasks and situations in which automation differs in reliability and may lead to more appropriate trust. The development of interventions can lead to calibrated trust in automation. © 2014, Human Factors and Ergonomics Society.

  8. Automation of dosimeters calibration for radiotherapy in secondary dosimetric calibration laboratory of the CPHR

    International Nuclear Information System (INIS)

    Acosta, Andy L. Romero; Lores, Stefan Gutierrez

    2013-01-01

    This paper presents the design and implementation of an automated system for measurements in the calibration of reference radiation dosimeters. It was made a software application that performs the acquisition of the measured values of electric charge, calculated calibration coefficient and automates the calibration certificate issuance. These values are stored in a log file on a PC. The use of the application improves control over the calibration process, helps to humanize the work and reduces personnel exposure. The tool developed has been applied to the calibration of dosimeters radiation patterns in the LSCD of the Centro de Proteccion e Higiene de las Radiaciones, Cuba

  9. Continuous Calibration of Trust in Automated Systems

    Science.gov (United States)

    2014-01-01

    Airlines Flight 214 in San Francisco. Therefore, understanding how users form, lose, and recover trust in imperfect automation is of critical...1997). Misuse and disuse can have fatal consequences; for example, inappropriate automation reliance has been implicated in the recent crash of Asiana

  10. Automated system for the calibration of magnetometers

    DEFF Research Database (Denmark)

    Petrucha, Vojtech; Kaspar, Petr; Ripka, Pavel

    2009-01-01

    A completely nonmagnetic calibration platform has been developed and constructed at DTU Space (Technical University of Denmark). It is intended for on-site scalar calibration of high-precise fluxgate magnetometers. An enhanced version of the same platform is being built at the Czech Technical Uni...... through custom-made optical incremental sensors. The system is controlled by a microcontroller, which executes commands from a computer. The properties of the system as well as calibration and measurement results will be presented. ©2009 American Institute of Physics...

  11. Automated intraoperative calibration for prostate cancer brachytherapy

    International Nuclear Information System (INIS)

    Kuiran Chen, Thomas; Heffter, Tamas; Lasso, Andras; Pinter, Csaba; Abolmaesumi, Purang; Burdette, E. Clif; Fichtinger, Gabor

    2011-01-01

    Purpose: Prostate cancer brachytherapy relies on an accurate spatial registration between the implant needles and the TRUS image, called ''calibration''. The authors propose a new device and a fast, automatic method to calibrate the brachytherapy system in the operating room, with instant error feedback. Methods: A device was CAD-designed and precision-engineered, which mechanically couples a calibration phantom with an exact replica of the standard brachytherapy template. From real-time TRUS images acquired from the calibration device and processed by the calibration system, the coordinate transformation between the brachytherapy template and the TRUS images was computed automatically. The system instantly generated a report of the target reconstruction accuracy based on the current calibration outcome. Results: Four types of validation tests were conducted. First, 50 independent, real-time calibration trials yielded an average of 0.57 ± 0.13 mm line reconstruction error (LRE) relative to ground truth. Second, the averaged LRE was 0.37 ± 0.25 mm relative to ground truth in tests with six different commercial TRUS scanners operating at similar imaging settings. Furthermore, testing with five different commercial stepper systems yielded an average of 0.29 ± 0.16 mm LRE relative to ground truth. Finally, the system achieved an average of 0.56 ± 0.27 mm target registration error (TRE) relative to ground truth in needle insertion tests through the template in a water tank. Conclusions: The proposed automatic, intraoperative calibration system for prostate cancer brachytherapy has achieved high accuracy, precision, and robustness.

  12. Hydrometer calibration by hydrostatic weighing with automated liquid surface positioning

    International Nuclear Information System (INIS)

    Aguilera, Jesus; Wright, John D; Bean, Vern E

    2008-01-01

    We describe an automated apparatus for calibrating hydrometers by hydrostatic weighing (Cuckow's method) in tridecane, a liquid of known, stable density, and with a relatively low surface tension and contact angle against glass. The apparatus uses a laser light sheet and a laser power meter to position the tridecane surface at the hydrometer scale mark to be calibrated with an uncertainty of 0.08 mm. The calibration results have an expanded uncertainty (with a coverage factor of 2) of 100 parts in 10 6 or less of the liquid density. We validated the apparatus by comparisons using water, toluene, tridecane and trichloroethylene, and found agreement within 40 parts in 10 6 or less. The new calibration method is consistent with earlier, manual calibrations performed by NIST. When customers use calibrated hydrometers, they may encounter uncertainties of 370 parts in 10 6 or larger due to surface tension, contact angle and temperature effects

  13. Hydrometer calibration by hydrostatic weighing with automated liquid surface positioning

    Science.gov (United States)

    Aguilera, Jesus; Wright, John D.; Bean, Vern E.

    2008-01-01

    We describe an automated apparatus for calibrating hydrometers by hydrostatic weighing (Cuckow's method) in tridecane, a liquid of known, stable density, and with a relatively low surface tension and contact angle against glass. The apparatus uses a laser light sheet and a laser power meter to position the tridecane surface at the hydrometer scale mark to be calibrated with an uncertainty of 0.08 mm. The calibration results have an expanded uncertainty (with a coverage factor of 2) of 100 parts in 106 or less of the liquid density. We validated the apparatus by comparisons using water, toluene, tridecane and trichloroethylene, and found agreement within 40 parts in 106 or less. The new calibration method is consistent with earlier, manual calibrations performed by NIST. When customers use calibrated hydrometers, they may encounter uncertainties of 370 parts in 106 or larger due to surface tension, contact angle and temperature effects.

  14. Automation of the Calibration of Reference Dosimeters Used in Radiotherapy

    International Nuclear Information System (INIS)

    Romero Acosta, A.; Gutierrez Lores, S.

    2013-01-01

    Traceability, accuracy and consistency of radiation measurements are essential in radiation dosimetry, particularly in radiotherapy, where the outcome of treatments is highly dependent on the radiation dose delivered to patients. The role of Secondary Standard Dosimetry Laboratories (SSDLs) is crucial in providing traceable calibrations to hospitals, since these laboratories disseminate calibrations at specific radiation qualities appropriate to the use of radiation measuring instruments. These laboratories follow IAEA/WHO guidelines for calibration procedures, often being current and charge measurements described in these guidelines a tedious task. However, these measurements are usually done using modern electrometers which are equipped with a RS-232 interface that allows instrument control from a PC. This paper presents the design and employment of an automated system aimed to the measurements of the radiotherapy dosimeters calibration process for Cobalt-60 gamma rays. A software was developed using Lab View, in order to achieve the acquisition of the charge values measured, calculation of the calibration coefficient and issue of a calibration certificate. A primary data report file is filled and stored in the PC's hard disk. By using this software tool, a better control over the calibration process is achieved, it reduces the need for human intervention and it also reduces the exposure of the laboratory staff. The automated system has been used for the calibration of reference dosimeters used in radiotherapy at the Cuban Secondary Standard Dosimetry Laboratory of the Center for Radiation Protection and Hygiene (Author)

  15. Automated system for the calibration of magnetometers

    International Nuclear Information System (INIS)

    Petrucha, Vojtech; Kaspar, Petr; Ripka, Pavel; Merayo, Jose M. G.

    2009-01-01

    A completely nonmagnetic calibration platform has been developed and constructed at DTU Space (Technical University of Denmark). It is intended for on-site scalar calibration of high-precise fluxgate magnetometers. An enhanced version of the same platform is being built at the Czech Technical University. There are three axes of rotation in this design (compared to two axes in the previous version). The addition of the third axis allows us to calibrate more complex devices. An electronic compass based on a vector fluxgate magnetometer and micro electro mechanical systems (MEMS) accelerometer is one example. The new platform can also be used to evaluate the parameters of the compass in all possible variations in azimuth, pitch, and roll. The system is based on piezoelectric motors, which are placed on a platform made of aluminum, brass, plastic, and glass. Position sensing is accomplished through custom-made optical incremental sensors. The system is controlled by a microcontroller, which executes commands from a computer. The properties of the system as well as calibration and measurement results will be presented

  16. A portable, automated, inexpensive mass and balance calibration system

    International Nuclear Information System (INIS)

    Maxwell, S.L. III; Clark, J.P.

    1987-01-01

    Reliable mass measurements are essential for a nuclear production facility or process control laboratory. DOE Order 5630.2 requires that traceable standards be used to calibrate and monitor equipment used for nuclear material measurements. To ensure the reliability of mass measurements and to comply with DOE traceable requirements, a portable, automated mass and balance calibration system is used at the Savannah River Plant. Automation is achieved using an EPSON HX-20 notebook computer, which can be operated via RS232C interfacing to electronic balances or function with manual data entry if computer interfacing is not feasible. This economical, comprehensive, user-friendly system has three main functions in a mass measurement control program (MMCP): balance certification, calibration of mass standards, and daily measurement of traceable standards. The balance certification program tests for accuracy, precision, sensitivity, linearity, and cornerloading versus specific requirements. The mass calibration program allows rapid calibration of inexpensive mass standards traceable to certified Class S standards. This MMCP permits daily measurement of traceable standards to monitor the reliability of balances during routine use. The automated system verifies balance calibration, stores results for future use, and provides a printed control chart of the stored data. Another feature of the system permits three different weighing routines that accommodate our need for varying degrees of reliability in routine weighing operations. 1 ref

  17. A portable, automated, inexpensive mass and balance calibration system

    International Nuclear Information System (INIS)

    Maxwell, S.L. III; Clark, J.P.

    1987-01-01

    Reliable mass measurements are essential for a nuclear production facility or process control laboratory. DOE Order 5630.2 requires that traceable standards be used to calibrate and monitor equipment used for nuclear material measurements. To ensure the reliability of mass measurements and to comply with DOE traceability requirements, a portable, automated mass and balance calibration system is used at the Savannah River Plant. Automation is achieved using an EPSON HX-20 notebook computer, which can be operated via RS232C interfacing to electronic balances or function with manual data entry if computer interfacing is not feasible. This economical, comprehensive, user-friendly system has three main functions in a mass measurement control program (MMCP): balance certification, calibration of mass standards, and daily measurement of traceable standards. The balance certification program tests for accuracy, precision, sensitivity, linearity, and cornerloading versus specific requirements. The mass calibration program allows rapid calibration of inexpensive mass standards traceable to certified Class S standards. This MMCP permits daily measurement of traceable standards to monitor the reliability of balances during routine use. The automated system verifies balance calibration, stores results for future use, and provides a printed control chart of the stored data. Another feature of the system permits three different weighing routines that accommodate their need for varying degrees of reliability in routine weighing operations

  18. Thrombin generation correlates with disease duration in multiple sclerosis (MS): Novel insights into the MS-associated prothrombotic state.

    Science.gov (United States)

    Parsons, Martin Em; O'Connell, Karen; Allen, Seamus; Egan, Karl; Szklanna, Paulina B; McGuigan, Christopher; Ní Áinle, Fionnuala; Maguire, Patricia B

    2017-01-01

    Thrombin is well recognised for its role in the coagulation cascade but it also plays a role in inflammation, with enhanced thrombin generation observed in several inflammatory disorders. Although patients with multiple sclerosis (MS) have a higher incidence of thrombotic disease, thrombin generation has not been studied to date. The aim of this study was to characterise calibrated automated thrombography parameters in patients with relapsing-remitting MS (RRMS) and primary progressive MS (PPMS) in comparison to healthy controls (HCs). Calibrated automated thrombography was performed on platelet poor plasma from 15 patients with RRMS, 15 with PPMS and 19 HCs. We found that patients with RRMS generate thrombin at a significantly faster rate than the less inflammatory subtype, PPMS or HCs. In addition, the speed of thrombin generation was significantly correlated with time from clinical diagnosis in both subtypes. However, in RRMS the rate of thrombin generation was increased with increased time from clinical diagnosis, while in PPMS the rate of thrombin generation decreased with increased time from clinical diagnosis. These data likely reflect the differential active proinflammatory states in each MS subtype and provide novel mechanistic insights into the clinically relevant prothrombotic state observed in these patients.

  19. Investigation of the thrombin-generating capacity, evaluated by thrombogram, and clot formation evaluated by thrombelastography of platelets stored in the blood bank for up to 7 days.

    Science.gov (United States)

    Johansson, P I; Svendsen, M S; Salado, J; Bochsen, L; Kristensen, A T

    2008-02-01

    Transfusion based on the Thrombelastograph (TEG) results reduces transfusion requirements in cardiac surgery and in liver transplantation. Taking the pivotal role of thrombin generation in the coagulation process into consideration, the clinical utility of the TEG may, in part, depend on its reflection of the dynamics of thrombin generation. The kinetics of thrombin generation of platelets stored for 2 and 7 days, respectively, was assessed by calibrated automated thrombogram (CAT) and the lag time (min), time to peak (ttPeak; min), peak (nm thrombin) and endogenous thrombin potential (ETP; nm thrombin*min) were registered. Clot formation was evaluated by TEG and the R time (min), maxial amplitude (MA; mm), time to maximum thrombus generation (TMG; min) and maximum thrombus generation (MTG; dynes cm(-2) s(-1)) and total thrombus generation (TTG; dyne cm(-2)) were registered. Platelets become more procoagulant, evaluated both by TEG and CAT during storage. The reduction in CAT lag time and the ttPeak correlated with a decrease in the TEG R time and TMG (P < 0.0001) as did the CAT peak thrombin generation and the TEG MTG (P = 0.0035). No correlation between ETP and TTG was found (P = 0.65). The kinetics of thrombin generation, as evaluated by CAT, correlates with the thrombus generation, as evaluated by thrombelastography and this may in part explain the clinical utility of the TEG in identifying clinically relevant coagulopathies, secondary to impaired thrombin generation.

  20. A New Automated Instrument Calibration Facility at the Savannah River Site

    International Nuclear Information System (INIS)

    Polz, E.; Rushton, R.O.; Wilkie, W.H.; Hancock, R.C.

    1998-01-01

    The Health Physics Instrument Calibration Facility at the Savannah River Site in Aiken, SC was expressly designed and built to calibrate portable radiation survey instruments. The facility incorporates recent advances in automation technology, building layout and construction, and computer software to improve the calibration process. Nine new calibration systems automate instrument calibration and data collection. The building is laid out so that instruments are moved from one area to another in a logical, efficient manner. New software and hardware integrate all functions such as shipping/receiving, work flow, calibration, testing, and report generation. Benefits include a streamlined and integrated program, improved efficiency, reduced errors, and better accuracy

  1. A direct thrombin inhibitor suppresses protein C activation and factor Va degradation in human plasma: Possible mechanisms of paradoxical enhancement of thrombin generation.

    Science.gov (United States)

    Kamisato, Chikako; Furugohri, Taketoshi; Morishima, Yoshiyuki

    2016-05-01

    We have demonstrated that antithrombin (AT)-independent thrombin inhibitors paradoxically increase thrombin generation (TG) in human plasma in a thrombomodulin (TM)- and protein C (PC)-dependent manner. We determined the effects of AT-independent thrombin inhibitors on the negative-feedback system, activation of PC and production and degradation of factor Va (FVa), as possible mechanisms underlying the paradoxical enhancement of TG. TG in human plasma containing 10nM TM was assayed by means of the calibrated automated thrombography. As an index of PC activation, plasma concentration of activated PC-PC inhibitor complex (aPC-PCI) was measured. The amounts of FVa heavy chain and its degradation product (FVa(307-506)) were examined by western blotting. AT-independent thrombin inhibitors, melagatran and dabigatran (both at 25-600nM) and 3-30μg/ml active site-blocked thrombin (IIai), increased peak levels of TG. Melagatran, dabigatran and IIai significantly decreased plasma concentration of aPC-PCI complex at 25nM or more, 75nM or more, and 10 and 30μg/ml, respectively. Melagatran (300nM) significantly increased FVa and decreased FVa(307-506). In contrast, a direct factor Xa inhibitor edoxaban preferentially inhibited thrombin generation (≥25nM), and higher concentrations were required to inhibit PC activation (≥150nM) and FVa degradation (300nM). The present study suggests that the inhibitions of protein C activation and subsequent degradation of FVa and increase in FVa by antithrombin-independent thrombin inhibitors may contribute to the paradoxical TG enhancement, and edoxaban may inhibit PC activation and FVa degradation as a result of TG suppression. Copyright © 2016 Elsevier Ltd. All rights reserved.

  2. Automation is an Effective Way to Improve Quality of Verification (Calibration) of Measuring Instruments

    Science.gov (United States)

    Golobokov, M.; Danilevich, S.

    2018-04-01

    In order to assess calibration reliability and automate such assessment, procedures for data collection and simulation study of thermal imager calibration procedure have been elaborated. The existing calibration techniques do not always provide high reliability. A new method for analyzing the existing calibration techniques and developing new efficient ones has been suggested and tested. A type of software has been studied that allows generating instrument calibration reports automatically, monitoring their proper configuration, processing measurement results and assessing instrument validity. The use of such software allows reducing man-hours spent on finalization of calibration data 2 to 5 times and eliminating a whole set of typical operator errors.

  3. Calibration-measurement unit for the automation of vector network analyzer measurements

    Directory of Open Access Journals (Sweden)

    I. Rolfes

    2008-05-01

    Full Text Available With the availability of multi-port vector network analyzers, the need for automated, calibrated measurement facilities increases. In this contribution, a calibration-measurement unit is presented which realizes a repeatable automated calibration of the measurement setup as well as a user-friendly measurement of the device under test (DUT. In difference to commercially available calibration units, which are connected to the ports of the vector network analyzer preceding a measurement and which are then removed so that the DUT can be connected, the presented calibration-measurement unit is permanently connected to the ports of the VNA for the calibration as well as for the measurement of the DUT. This helps to simplify the calibrated measurement of complex scattering parameters. Moreover, a full integration of the calibration unit into the analyzer setup becomes possible. The calibration-measurement unit is based on a multiport switch setup of e.g. electromechanical relays. Under the assumption of symmetry of a switch, on the one hand the unit realizes the connection of calibration standards like one-port reflection standards and two-port through connections between different ports and on the other hand it enables the connection of the DUT. The calibration-measurement unit is applicable for two-port VNAs as well as for multiport VNAs. For the calibration of the unit, methods with completely known calibration standards like SOLT (short, open, load, through as well as self-calibration procedures like TMR or TLR can be applied.

  4. Automated system for calibration and control of the CHSPP-800 multichannel γ detector parameters

    International Nuclear Information System (INIS)

    Avvakumov, N.A.; Belikov, N.I.; Goncharenko, Yu.M.

    1987-01-01

    An automated system for adjustment, calibration and control of total absorption Cherenkov spectrometer is described. The system comprises a mechanical platform, capable of moving in two mutually perpendicular directions; movement detectors and limit switches; power unit, automation unit with remote control board. The automated system can operate both in manual control regime with coordinate control by a digital indicator, and in operation regime with computer according to special programs. The platform mounting accuracy is ± 0.1 mm. Application of the automated system has increased the rate of the course of the counter adjustment works 3-5 times

  5. BRAIN initiative: transcranial magnetic stimulation automation and calibration.

    Science.gov (United States)

    Todd, Garth D; Abdellatif, Ahmed; Sabouni, Abas

    2014-01-01

    In this paper, we introduced an automated TMS system with robot control and optical sensor combined with neuronavigation software. By using the robot, the TMS coil can be accurately positioned over any preselected brain region. The neuronavigation system provides an accurate positioning of a magnetic coil in order to induce a specific cortical excitation. An infrared optical measurement device is also used in order to detect and compensate for head movements of the patient. This procedure was simulated using a PC based robotic simulation program. The proposed automated robot system is integrated with TMS numerical solver and allows users to actually see the depth, location, and shape of the induced eddy current on the computer monitor.

  6. Extracellular histones promote thrombin generation through platelet-dependent mechanisms: involvement of platelet TLR2 and TLR4

    Science.gov (United States)

    Semeraro, Fabrizio; Ammollo, Concetta T.; Morrissey, James H.; Dale, George L.; Friese, Paul; Esmon, Naomi L.

    2011-01-01

    The release of histones from dying cells is associated with microvascular thrombosis and, because histones activate platelets, this could represent a possible pathogenic mechanism. In the present study, we assessed the influence of histones on the procoagulant potential of human platelets in platelet-rich plasma (PRP) and in purified systems. Histones dose-dependently enhanced thrombin generation in PRP in the absence of any trigger, as evaluated by calibrated automated thrombinography regardless of whether the contact phase was inhibited. Activation of coagulation required the presence of fully activatable platelets and was not ascribable to platelet tissue factor, whereas targeting polyphosphate with phosphatase reduced thrombin generation even when factor XII (FXII) was blocked or absent. In the presence of histones, purified polyphosphate was able to induce thrombin generation in plasma independently of FXII. In purified systems, histones induced platelet aggregation; P-selectin, phosphatidylserine, and FV/Va expression; and prothrombinase activity. Blocking platelet TLR2 and TLR4 with mAbs reduced the percentage of activated platelets and lowered the amount of thrombin generated in PRP. These data show that histone-activated platelets possess a procoagulant phenotype that drives plasma thrombin generation and suggest that TLR2 and TLR4 mediate the activation process. PMID:21673343

  7. Experiences in Automated Calibration of a Nickel Equation of State

    Science.gov (United States)

    Carpenter, John H.

    2017-06-01

    Wide availability of large computers has led to increasing incorporation of computational data, such as from density functional theory molecular dynamics, in the development of equation of state (EOS) models. Once a grid of computational data is available, it is usually left to an expert modeler to model the EOS using traditional techniques. One can envision the possibility of using the increasing computing resources to perform black-box calibration of EOS models, with the goal of reducing the workload on the modeler or enabling non-experts to generate good EOSs with such a tool. Progress towards building such a black-box calibration tool will be explored in the context of developing a new, wide-range EOS for nickel. While some details of the model and data will be shared, the focus will be on what was learned by automatically calibrating the model in a black-box method. Model choices and ensuring physicality will also be discussed. Sandia National Laboratories is a multi-mission laboratory managed and operated by Sandia Corporation, a wholly owned subsidiary of Lockheed Martin Corporation, for the U.S. Department of Energy's National Nuclear Security Administration under contract DE-AC04-94AL85000.

  8. Protein Z efficiently depletes thrombin generation in disseminated intravascular coagulation with poor prognosis.

    Science.gov (United States)

    Lee, Nuri; Kim, Ji-Eun; Gu, Ja-Yoon; Yoo, Hyun Ju; Kim, Inho; Yoon, Sung-Soo; Park, Seonyang; Han, Kyou-Sup; Kim, Hyun Kyung

    2016-01-01

    Disseminated intravascular coagulation (DIC) is characterized by consumption of coagulation factors and anticoagulants. Thrombin generation assay (TGA) gives useful information about global hemostatic status. We developed a new TGA system that anticoagulant addition can deplete thrombin generation in plasma, which may reflect defective anticoagulant system in DIC. TGAs were measured on the calibrated automated thrombogram with and without thrombomodulin or protein Z in 152 patients who were suspected of having DIC, yielding four parameters including lag time, endogenous thrombin potential, peak thrombin and time-to-peak in each experiment. Nonsurvivors showed significantly prolonged lag time and time-to-peak in TGA-protein Z system, which was performed with added protein Z. In multivariate Cox regression analysis, lag time and time-to-peak in TGA system were significant independent prognostic factors. In TGA-protein Z system, lag time and time-to-peak were revealed as independent prognostic factors of DIC. Protein Z addition could potentiate its anticoagulant effect in DIC with poor prognosis, suggesting the presence of defective protein Z system. The prolonged lag time and time-to-peak in both TGA and TGA-protein Z systems are expected to be used as independent prognostic factors of DIC.

  9. Automated gauge block pair length difference calibration and associated uncertainty sources

    International Nuclear Information System (INIS)

    Oliveira, W Jr; França, R S

    2015-01-01

    A reduction for interferometric uncertainties in length difference at gauge block pairs is presented. An automated processing designed to compensate geometric fringe visualization effects and four-alternate wringing technique are used to achieve small combined uncertainties for length difference calibrations, maintaining a good compliance with the EAL-G21 determinations. (paper)

  10. GESCAL: Quality management automated system for a calibration and test laboratory

    International Nuclear Information System (INIS)

    Manzano de Armas, J.; Valdes Ramos, M.; Morales Monzon, J.A.

    1998-01-01

    GESCAL is a software created to automate all elements composing the quality system in a calibration and test laboratory. It also evaluates quality according to its objectives and policies. This integrated data system decreases considerably the amount of time devoted to manage quality. It is speedier in searching and evaluating information registers thus notably in reducing the workload for laboratory staff

  11. ISS Payload Racks Automated Flow Control Calibration Method

    Science.gov (United States)

    Simmonds, Boris G.

    2003-01-01

    Payload Racks utilize MTL and/or LTL station water for cooling of payloads and avionics. Flow control range from valves of fully closed, to up to 300 Ibmhr. Instrument accuracies are as high as f 7.5 Ibm/hr for flow sensors and f 3 Ibm/hr for valve controller, for a total system accuracy of f 10.5 Ibm/hr. Improved methodology was developed, tested and proven that reduces accuracy of the commanded flows to less than f 1 Ibmhr. Uethodology could be packed in a "calibration kit" for on- orbit flow sensor checkout and recalibration, extending the rack operations before return to earth. -

  12. A new method for automated dynamic calibration of tipping-bucket rain gauges

    Science.gov (United States)

    Humphrey, M.D.; Istok, J.D.; Lee, J.Y.; Hevesi, J.A.; Flint, A.L.

    1997-01-01

    Existing methods for dynamic calibration of tipping-bucket rain gauges (TBRs) can be time consuming and labor intensive. A new automated dynamic calibration system has been developed to calibrate TBRs with minimal effort. The system consists of a programmable pump, datalogger, digital balance, and computer. Calibration is performed in two steps: 1) pump calibration and 2) rain gauge calibration. Pump calibration ensures precise control of water flow rates delivered to the rain gauge funnel; rain gauge calibration ensures precise conversion of bucket tip times to actual rainfall rates. Calibration of the pump and one rain gauge for 10 selected pump rates typically requires about 8 h. Data files generated during rain gauge calibration are used to compute rainfall intensities and amounts from a record of bucket tip times collected in the field. The system was tested using 5 types of commercial TBRs (15.2-, 20.3-, and 30.5-cm diameters; 0.1-, 0.2-, and 1.0-mm resolutions) and using 14 TBRs of a single type (20.3-cm diameter; 0.1-mm resolution). Ten pump rates ranging from 3 to 154 mL min-1 were used to calibrate the TBRs and represented rainfall rates between 6 and 254 mm h-1 depending on the rain gauge diameter. All pump calibration results were very linear with R2 values greater than 0.99. All rain gauges exhibited large nonlinear underestimation errors (between 5% and 29%) that decreased with increasing rain gauge resolution and increased with increasing rainfall rate, especially for rates greater than 50 mm h-1. Calibration curves of bucket tip time against the reciprocal of the true pump rate for all rain gauges also were linear with R2 values of 0.99. Calibration data for the 14 rain gauges of the same type were very similar, as indicated by slope values that were within 14% of each other and ranged from about 367 to 417 s mm h-1. The developed system can calibrate TBRs efficiently, accurately, and virtually unattended and could be modified for use with other

  13. Automated calibration of TECAN genesis liquid handling workstation utilizing an online balance and density meter.

    Science.gov (United States)

    Xie, Iris H; Wang, Michael H; Carpenter, Richard; Wu, Henry Y

    2004-02-01

    With robotics widely used in bioanalytical assays, accurate system performance is essential to ensure the quality and productivity of the robotics. In our lab, an automated calibration procedure has been developed to evaluate the precision and accuracy of the TECAN (Research Triangle Park, NC, U.S.A.) Genesis liquid handling system in a bioanalytical laboratory setting. The calibrations were performed by transferring and weighing the solvents automatically on a microbalance controlled by a Gemini program. From the data acquired, calibration reports were generated using a template. The novel aspect of this approach is the use of an on-line balance and a density meter, both of which combine to make the calibration process simple, efficient, and precise. For quantitative bioanalysis, a variety of solvents, including methanol, water, mixed solvents, and plasma, are typically used to prepare standards and unknown samples. Density information is usually unknown for the mixed solvents, and the density of plasma can vary from species to species. However, with the use of a universal density meter, the density could be obtained in seconds. The issue of solvent evaporation during the calibration process was also addressed. Calibration curves were set up for various liquid classes. Pipetting volumes ranged from 10 microL to 900 microL. Precision and accuracy results obtained from the semiannual performance evaluations showed this procedure to be reliable and user-friendly. Using the automated calibration procedure, the calibration and performance evaluation of the robotic system is considerably more efficient, and the incidence of unacceptable precision and accuracy is greatly reduced.

  14. Shared robotic system: automated pipette calibration and pipette tip filter assembly

    International Nuclear Information System (INIS)

    Addison, J.H. Jr.; Dyches, G.M.

    1985-01-01

    At the Savannah River Laboratory a Zymate Laboratory Automation System has been developed to perform two completely independent tasks within one work cell. One operation is the precise calibration of pipettes; the other is the assembly of a filter in a pipette tip. Since neither task requires full robot time, the shared system is an economical means of robotizing both processes. These are tedious, repetitive, time consuming tasks; and human operators fail to yield constant results. Automation insures a repeatable process which increases product quality

  15. Enhancing Seismic Calibration Research Through Software Automation and Scientific Information Management

    Energy Technology Data Exchange (ETDEWEB)

    Ruppert, S D; Dodge, D A; Ganzberger, M D; Harris, D B; Hauk, T F

    2009-07-07

    The National Nuclear Security Administration (NNSA) Ground-Based Nuclear Explosion Monitoring Research and Development (GNEMRD) Program at LLNL continues to make significant progress enhancing the process of deriving seismic calibrations and performing scientific integration, analysis, and information management with software automation tools. Our tool efforts address the problematic issues of very large datasets and varied formats encountered during seismic calibration research. New information management and analysis tools have resulted in demonstrated gains in efficiency of producing scientific data products and improved accuracy of derived seismic calibrations. In contrast to previous years, software development work this past year has emphasized development of automation at the data ingestion level. This change reflects a gradually-changing emphasis in our program from processing a few large data sets that result in a single integrated delivery, to processing many different data sets from a variety of sources. The increase in the number of sources had resulted in a large increase in the amount of metadata relative to the final volume of research products. Software developed this year addresses the problems of: (1) Efficient metadata ingestion and conflict resolution; (2) Automated ingestion of bulletin information; (3) Automated ingestion of waveform information from global data centers; and (4) Site Metadata and Response transformation required for certain products. This year, we also made a significant step forward in meeting a long-standing goal of developing and using a waveform correlation framework. Our objective for such a framework is to extract additional calibration data (e.g. mining blasts) and to study the extent to which correlated seismicity can be found in global and regional scale environments.

  16. Automated magnification calibration in transmission electron microscopy using Fourier analysis of replica images

    International Nuclear Information System (INIS)

    Laak, Jeroen A.W.M. van der; Dijkman, Henry B.P.M.; Pahlplatz, Martin M.M.

    2006-01-01

    The magnification factor in transmission electron microscopy is not very precise, hampering for instance quantitative analysis of specimens. Calibration of the magnification is usually performed interactively using replica specimens, containing line or grating patterns with known spacing. In the present study, a procedure is described for automated magnification calibration using digital images of a line replica. This procedure is based on analysis of the power spectrum of Fourier transformed replica images, and is compared to interactive measurement in the same images. Images were used with magnification ranging from 1,000x to 200,000x. The automated procedure deviated on average 0.10% from interactive measurements. Especially for catalase replicas, the coefficient of variation of automated measurement was considerably smaller (average 0.28%) compared to that of interactive measurement (average 3.5%). In conclusion, calibration of the magnification in digital images from transmission electron microscopy may be performed automatically, using the procedure presented here, with high precision and accuracy

  17. Evaluation of Automated Model Calibration Techniques for Residential Building Energy Simulation

    Energy Technology Data Exchange (ETDEWEB)

    and Ben Polly, Joseph Robertson [National Renewable Energy Lab. (NREL), Golden, CO (United States); Polly, Ben [National Renewable Energy Lab. (NREL), Golden, CO (United States); Collis, Jon [Colorado School of Mines, Golden, CO (United States)

    2013-09-01

    This simulation study adapts and applies the general framework described in BESTEST-EX (Judkoff et al 2010) for self-testing residential building energy model calibration methods. BEopt/DOE-2.2 is used to evaluate four mathematical calibration methods in the context of monthly, daily, and hourly synthetic utility data for a 1960's-era existing home in a cooling-dominated climate. The home's model inputs are assigned probability distributions representing uncertainty ranges, random selections are made from the uncertainty ranges to define "explicit" input values, and synthetic utility billing data are generated using the explicit input values. The four calibration methods evaluated in this study are: an ASHRAE 1051-RP-based approach (Reddy and Maor 2006), a simplified simulated annealing optimization approach, a regression metamodeling optimization approach, and a simple output ratio calibration approach. The calibration methods are evaluated for monthly, daily, and hourly cases; various retrofit measures are applied to the calibrated models and the methods are evaluated based on the accuracy of predicted savings, computational cost, repeatability, automation, and ease of implementation.

  18. Evaluation of Automated Model Calibration Techniques for Residential Building Energy Simulation

    Energy Technology Data Exchange (ETDEWEB)

    Robertson, J.; Polly, B.; Collis, J.

    2013-09-01

    This simulation study adapts and applies the general framework described in BESTEST-EX (Judkoff et al 2010) for self-testing residential building energy model calibration methods. BEopt/DOE-2.2 is used to evaluate four mathematical calibration methods in the context of monthly, daily, and hourly synthetic utility data for a 1960's-era existing home in a cooling-dominated climate. The home's model inputs are assigned probability distributions representing uncertainty ranges, random selections are made from the uncertainty ranges to define 'explicit' input values, and synthetic utility billing data are generated using the explicit input values. The four calibration methods evaluated in this study are: an ASHRAE 1051-RP-based approach (Reddy and Maor 2006), a simplified simulated annealing optimization approach, a regression metamodeling optimization approach, and a simple output ratio calibration approach. The calibration methods are evaluated for monthly, daily, and hourly cases; various retrofit measures are applied to the calibrated models and the methods are evaluated based on the accuracy of predicted savings, computational cost, repeatability, automation, and ease of implementation.

  19. Construction and calibration of a low cost and fully automated vibrating sample magnetometer

    International Nuclear Information System (INIS)

    El-Alaily, T.M.; El-Nimr, M.K.; Saafan, S.A.; Kamel, M.M.; Meaz, T.M.; Assar, S.T.

    2015-01-01

    A low cost vibrating sample magnetometer (VSM) has been constructed by using an electromagnet and an audio loud speaker; where both are controlled by a data acquisition device. The constructed VSM records the magnetic hysteresis loop up to 8.3 KG at room temperature. The apparatus has been calibrated and tested by using magnetic hysteresis data of some ferrite samples measured by two scientifically calibrated magnetometers; model (Lake Shore 7410) and model (LDJ Electronics Inc. Troy, MI). Our VSM lab-built new design proved success and reliability. - Highlights: • A low cost automated vibrating sample magnetometer VSM has been constructed. • The VSM records the magnetic hysteresis loop up to 8.3 KG at room temperature. • The VSM has been calibrated and tested by using some measured ferrite samples. • Our VSM lab-built new design proved success and reliability

  20. Construction and calibration of a low cost and fully automated vibrating sample magnetometer

    Energy Technology Data Exchange (ETDEWEB)

    El-Alaily, T.M., E-mail: toson_alaily@yahoo.com [Physics Department, Faculty of Science, Tanta University, Tanta (Egypt); El-Nimr, M.K.; Saafan, S.A.; Kamel, M.M.; Meaz, T.M. [Physics Department, Faculty of Science, Tanta University, Tanta (Egypt); Assar, S.T. [Engineering Physics and Mathematics Department, Faculty of Engineering, Tanta University, Tanta (Egypt)

    2015-07-15

    A low cost vibrating sample magnetometer (VSM) has been constructed by using an electromagnet and an audio loud speaker; where both are controlled by a data acquisition device. The constructed VSM records the magnetic hysteresis loop up to 8.3 KG at room temperature. The apparatus has been calibrated and tested by using magnetic hysteresis data of some ferrite samples measured by two scientifically calibrated magnetometers; model (Lake Shore 7410) and model (LDJ Electronics Inc. Troy, MI). Our VSM lab-built new design proved success and reliability. - Highlights: • A low cost automated vibrating sample magnetometer VSM has been constructed. • The VSM records the magnetic hysteresis loop up to 8.3 KG at room temperature. • The VSM has been calibrated and tested by using some measured ferrite samples. • Our VSM lab-built new design proved success and reliability.

  1. Automation of RELAP5 input calibration and code validation using genetic algorithm

    International Nuclear Information System (INIS)

    Phung, Viet-Anh; Kööp, Kaspar; Grishchenko, Dmitry; Vorobyev, Yury; Kudinov, Pavel

    2016-01-01

    Highlights: • Automated input calibration and code validation using genetic algorithm is presented. • Predictions generally overlap experiments for individual system response quantities (SRQs). • It was not possible to predict simultaneously experimental maximum flow rate and oscillation period. • Simultaneous consideration of multiple SRQs is important for code validation. - Abstract: Validation of system thermal-hydraulic codes is an important step in application of the codes to reactor safety analysis. The goal of the validation process is to determine how well a code can represent physical reality. This is achieved by comparing predicted and experimental system response quantities (SRQs) taking into account experimental and modelling uncertainties. Parameters which are required for the code input but not measured directly in the experiment can become an important source of uncertainty in the code validation process. Quantification of such parameters is often called input calibration. Calibration and uncertainty quantification may become challenging tasks when the number of calibrated input parameters and SRQs is large and dependencies between them are complex. If only engineering judgment is employed in the process, the outcome can be prone to so called “user effects”. The goal of this work is to develop an automated approach to input calibration and RELAP5 code validation against data on two-phase natural circulation flow instability. Multiple SRQs are used in both calibration and validation. In the input calibration, we used genetic algorithm (GA), a heuristic global optimization method, in order to minimize the discrepancy between experimental and simulation data by identifying optimal combinations of uncertain input parameters in the calibration process. We demonstrate the importance of the proper selection of SRQs and respective normalization and weighting factors in the fitness function. In the code validation, we used maximum flow rate as the

  2. Automation of RELAP5 input calibration and code validation using genetic algorithm

    Energy Technology Data Exchange (ETDEWEB)

    Phung, Viet-Anh, E-mail: vaphung@kth.se [Division of Nuclear Power Safety, Royal Institute of Technology, Roslagstullsbacken 21, 10691 Stockholm (Sweden); Kööp, Kaspar, E-mail: kaspar@safety.sci.kth.se [Division of Nuclear Power Safety, Royal Institute of Technology, Roslagstullsbacken 21, 10691 Stockholm (Sweden); Grishchenko, Dmitry, E-mail: dmitry@safety.sci.kth.se [Division of Nuclear Power Safety, Royal Institute of Technology, Roslagstullsbacken 21, 10691 Stockholm (Sweden); Vorobyev, Yury, E-mail: yura3510@gmail.com [National Research Center “Kurchatov Institute”, Kurchatov square 1, Moscow 123182 (Russian Federation); Kudinov, Pavel, E-mail: pavel@safety.sci.kth.se [Division of Nuclear Power Safety, Royal Institute of Technology, Roslagstullsbacken 21, 10691 Stockholm (Sweden)

    2016-04-15

    Highlights: • Automated input calibration and code validation using genetic algorithm is presented. • Predictions generally overlap experiments for individual system response quantities (SRQs). • It was not possible to predict simultaneously experimental maximum flow rate and oscillation period. • Simultaneous consideration of multiple SRQs is important for code validation. - Abstract: Validation of system thermal-hydraulic codes is an important step in application of the codes to reactor safety analysis. The goal of the validation process is to determine how well a code can represent physical reality. This is achieved by comparing predicted and experimental system response quantities (SRQs) taking into account experimental and modelling uncertainties. Parameters which are required for the code input but not measured directly in the experiment can become an important source of uncertainty in the code validation process. Quantification of such parameters is often called input calibration. Calibration and uncertainty quantification may become challenging tasks when the number of calibrated input parameters and SRQs is large and dependencies between them are complex. If only engineering judgment is employed in the process, the outcome can be prone to so called “user effects”. The goal of this work is to develop an automated approach to input calibration and RELAP5 code validation against data on two-phase natural circulation flow instability. Multiple SRQs are used in both calibration and validation. In the input calibration, we used genetic algorithm (GA), a heuristic global optimization method, in order to minimize the discrepancy between experimental and simulation data by identifying optimal combinations of uncertain input parameters in the calibration process. We demonstrate the importance of the proper selection of SRQs and respective normalization and weighting factors in the fitness function. In the code validation, we used maximum flow rate as the

  3. A method for automating calibration and records management for instrumentation and dosimetry

    International Nuclear Information System (INIS)

    O'Brien, J.M. Jr.; Rushton, R.O.; Burns, R.E. Jr.

    1993-01-01

    Current industry requirements are becoming more stringent on quality assurance records and documentation for calibration of instruments and dosimetry. A novel method is presented here that will allow a progressive automation scheme to be used in pursuit of that goal. This concept is based on computer-controlled irradiators that can act as stand-alone devices or be interfaced to other components via a computer local area network. In this way, complete systems can be built with modules to create a records management system to meet the needs of small laboratories or large multi-building calibration groups. Different database engines or formats can be used simply by replacing a module. Modules for temperature and pressure monitoring or shipping and receiving can be added, as well as equipment modules for direct IEEE-488 interface to electrometers and other instrumentation

  4. A method for automating calibration and records management for instrumentation and dosimetry

    Energy Technology Data Exchange (ETDEWEB)

    O`Brien, J.M. Jr.; Rushton, R.O.; Burns, R.E. Jr. [Atlan-Tech, Inc., Roswell, GA (United States)

    1993-12-31

    Current industry requirements are becoming more stringent on quality assurance records and documentation for calibration of instruments and dosimetry. A novel method is presented here that will allow a progressive automation scheme to be used in pursuit of that goal. This concept is based on computer-controlled irradiators that can act as stand-alone devices or be interfaced to other components via a computer local area network. In this way, complete systems can be built with modules to create a records management system to meet the needs of small laboratories or large multi-building calibration groups. Different database engines or formats can be used simply by replacing a module. Modules for temperature and pressure monitoring or shipping and receiving can be added, as well as equipment modules for direct IEEE-488 interface to electrometers and other instrumentation.

  5. The standard calibration instrument automation system for the atomic absorption spectrophotometer. Part 3: Program documentation

    Science.gov (United States)

    Ryan, D. P.; Roth, G. S.

    1982-04-01

    Complete documentation of the 15 programs and 11 data files of the EPA Atomic Absorption Instrument Automation System is presented. The system incorporates the following major features: (1) multipoint calibration using first, second, or third degree regression or linear interpolation, (2) timely quality control assessments for spiked samples, duplicates, laboratory control standards, reagent blanks, and instrument check standards, (3) reagent blank subtraction, and (4) plotting of calibration curves and raw data peaks. The programs of this system are written in Data General Extended BASIC, Revision 4.3, as enhanced for multi-user, real-time data acquisition. They run in a Data General Nova 840 minicomputer under the operating system RDOS, Revision 6.2. There is a functional description, a symbol definitions table, a functional flowchart, a program listing, and a symbol cross reference table for each program. The structure of every data file is also detailed.

  6. Analysis and Automation of Calibration Process for Measurement Coils for Particle Accelerator Magnets

    CERN Document Server

    AUTHOR|(CDS)2226624; Azzam, Jamal; Chanthery, Elodie

    Various techniques are used to measure magnetic fields within the Magnetic Measurement (MM) section, all of which require the use of specific sensors. These sensors need to be calibrated in order to map the resulting signals to the characteristics of the magnetic field. Thus, the calibration procedure would benefit from being improved and made more stable, efficient, and technologically up-to-date. Consequently, to improve the efficiency of the calibration procedure and the work processes it was first necessary to analyze them and identify potential issues and weaknesses to be further addressed. The calibration procedure mainly suffered from too many manual operations, which were potential sources of error, and from outdated readout and data acquisition software and hardware. Firstly, a new software program had to be developed using a C++ framework called Flexible Framework for Magnetic Measurements (FFMM), that would automate the data acquisition and analysis. Next, a feasibility study had to be done ...

  7. Comparatively Studied Color Correction Methods for Color Calibration of Automated Microscopy Complex of Biomedical Specimens

    Directory of Open Access Journals (Sweden)

    T. A. Kravtsova

    2016-01-01

    Full Text Available The paper considers a task of generating the requirements and creating a calibration target for automated microscopy systems (AMS of biomedical specimens to provide the invariance of algorithms and software to the hardware configuration. The required number of color fields of the calibration target and their color coordinates are mostly determined by the color correction method, for which coefficients of the equations are estimated during the calibration process. The paper analyses existing color calibration techniques for digital imaging systems using an optical microscope and shows that there is a lack of published results of comparative studies to demonstrate a particular useful color correction method for microscopic images. A comparative study of ten image color correction methods in RGB space using polynomials and combinations of color coordinate of different orders was carried out. The method of conditioned least squares to estimate the coefficients in the color correction equations using captured images of 217 color fields of the calibration target Kodak Q60-E3 was applied. The regularization parameter in this method was chosen experimentally. It was demonstrated that the best color correction quality characteristics are provided by the method that uses a combination of color coordinates of the 3rd order. The study of the influence of the number and the set of color fields included in calibration target on color correction quality for microscopic images was performed. Six train sets containing 30, 35, 40, 50, 60 and 80 color fields, and test set of 47 color fields not included in any of the train sets were formed. It was found out that the train set of 60 color fields minimizes the color correction error values for both operating modes of digital camera: using "default" color settings and with automatic white balance. At the same time it was established that the use of color fields from the widely used now Kodak Q60-E3 target does not

  8. Implication of Free Fatty Acids in Thrombin Generation and Fibrinolysis in Vascular Inflammation in Zucker Rats and Evolution with Aging

    Directory of Open Access Journals (Sweden)

    Jérémy Lagrange

    2017-11-01

    Full Text Available Background: The metabolic syndrome (MetS and aging are associated with modifications in blood coagulation factors, vascular inflammation, and increased risk of thrombosis.Objectives: Our aim was to determine concomitant changes in thrombin generation in the blood compartment and at the surface of vascular smooth muscle cells (VSMCs and its interplay with adipokines, free fatty acids (FFA, and metalloproteinases (MMPs in obese Zucker rats that share features of the human MetS.Methods: Obese and age-matched lean Zucker rats were compared at 25 and 80 weeks of age. Thrombin generation was assessed by calibrated automated thrombography (CAT.Results: Endogenous thrombin potential (ETP was increased in obese rats independent of platelets and age. Clot half-lysis time was delayed with obesity and age. Interleukin (IL-1β and IL-13 were increased with obesity and age respectively. Addition of exogenous fibrinogen, leptin, linoleic, or palmitic acid increased thrombin generation in plasma whereas adiponectin had an opposite effect. ETP was increased at the surface of VSMCs from obese rats and addition of exogenous palmitic acid further enhanced ETP values. Gelatinase activity was increased in aorta at both ages in obese rats and MMP-2 activity was increased in VSMCs from obese rats.Conclusions: Our study demonstrated in MetS an early prothrombotic phenotype of the blood compartment reinforced by procoagulant properties of dedifferentiated and inflammatory VSMCs. Mechanisms involved (1 increased fibrinogen and impaired fibrinolysis and (2 increased saturated fatty acids responsible for additive procoagulant effects. Whether specifically targeting this hypercoagulability using direct thrombin inhibitors would improve outcome in MetS is worth investigating.

  9. Automated Energy Calibration and Fitting of LaCl3(Ce y-Spectra Using Peak Likelihood and Tabu Search

    Directory of Open Access Journals (Sweden)

    Timothy P. McClanahan

    2008-10-01

    Full Text Available An automated method for ?-emission spectrum calibration and deconvolution is presented for spaceflight applications for a Cerium doped Lanthanum Chloride, (LaCl3(Ce ?-ray detector system. This detector will be coupled with a pulsed neutron generator (PNG to induce and enhance nuclide signal quality and rates, yielding large volumes of spectral information. Automated analytical methods are required to deconvolve and quantify nuclide signals from spectra; this will both reduce human interactions in spectrum analysis and facilitate feedback to automated robotic and operations planning. Initial system tests indicate significant energy calibration drifts (>6%, that which must be mitigated for spectrum analysis. A linear energy calibration model is presently considered, with gain and zero factors. Deconvolution methods incorporate a tabu search heuristic to formulate and optimize searches using memory structures. Iterative use of a peak likelihood methodology identifies global calibration minima and peak areas. The method is compared to manual methods of calibration and indicates superior performance using tabu methods. Performance of the Tabu enhanced calibration method is superior to similar unoptimized local search. The techniques are also applicable to other emission spectroscopy, eg. X-ray and neutron.

  10. Automated workflows for critical time-dependent calibrations at the CMS experiment.

    CERN Document Server

    Cerminara, Gianluca

    2015-01-01

    Fast and efficient methods for the calibration and the alignment ofthe detector are a key asset to exploit the physics potential of theCompact Muon Solenoid (CMS) detector and to ensure timely preparationof results for conferences and publications.To achieve this goal, the CMS experiment has set up a powerfulframework. This includes automated workflows in the context of a promptcalibration concept, which allows for a quick turnaround of thecalibration process following as fast as possible any change inrunning conditions.The presentation will review the design and operational experience ofthese workflows and the related monitoring system during the LHC RunIand focus on the development, deployment and commissioning in preparation of RunII.

  11. An automated calibration laboratory for flight research instrumentation: Requirements and a proposed design approach

    Science.gov (United States)

    Oneill-Rood, Nora; Glover, Richard D.

    1990-01-01

    NASA's Dryden Flight Research Facility (Ames-Dryden), operates a diverse fleet of research aircraft which are heavily instrumented to provide both real time data for in-flight monitoring and recorded data for postflight analysis. Ames-Dryden's existing automated calibration (AUTOCAL) laboratory is a computerized facility which tests aircraft sensors to certify accuracy for anticipated harsh flight environments. Recently, a major AUTOCAL lab upgrade was initiated; the goal of this modernization is to enhance productivity and improve configuration management for both software and test data. The new system will have multiple testing stations employing distributed processing linked by a local area network to a centralized database. The baseline requirements for the new AUTOCAL lab and the design approach being taken for its mechanization are described.

  12. Calibration

    International Nuclear Information System (INIS)

    Greacen, E.L.; Correll, R.L.; Cunningham, R.B.; Johns, G.G.; Nicolls, K.D.

    1981-01-01

    Procedures common to different methods of calibration of neutron moisture meters are outlined and laboratory and field calibration methods compared. Gross errors which arise from faulty calibration techniques are described. The count rate can be affected by the dry bulk density of the soil, the volumetric content of constitutional hydrogen and other chemical components of the soil and soil solution. Calibration is further complicated by the fact that the neutron meter responds more strongly to the soil properties close to the detector and source. The differences in slope of calibration curves for different soils can be as much as 40%

  13. ENHANCING SEISMIC CALIBRATION RESEARCH THROUGH SOFTWARE AUTOMATION AND SCIENTIFIC INFORMATION MANAGEMENT

    Energy Technology Data Exchange (ETDEWEB)

    Ruppert, S; Dodge, D A; Ganzberger, M D; Hauk, T F; Matzel, E M

    2008-07-03

    The National Nuclear Security Administration (NNSA) Ground-Based Nuclear Explosion Monitoring Research and Development (GNEMRD) Program at LLNL continues to make significant progress enhancing the process of deriving seismic calibrations and performing scientific integration, analysis, and information management with software automation tools. Our tool efforts address the problematic issues of very large datasets and varied formats encountered during seismic calibration research. New information management and analysis tools have resulted in demonstrated gains in efficiency of producing scientific data products and improved accuracy of derived seismic calibrations. The foundation of a robust, efficient data development and processing environment is comprised of many components built upon engineered versatile libraries. We incorporate proven industry 'best practices' throughout our code and apply source code and bug tracking management as well as automatic generation and execution of unit tests for our experimental, development and production lines. Significant software engineering and development efforts have produced an object-oriented framework that provides database centric coordination between scientific tools, users, and data. Over a half billion parameters, signals, measurements, and metadata entries are all stored in a relational database accessed by an extensive object-oriented multi-technology software framework that includes stored procedures, real-time transactional database triggers and constraints, as well as coupled Java and C++ software libraries to handle the information interchange and validation requirements. Significant resources were applied to schema design to enable management of processing methods and station parameters, responses and metadata. This allowed for the development of merged ground-truth (GT) data sets compiled by the NNSA labs and AFTAC that include hundreds of thousands of events and tens of millions of arrivals. The

  14. Automation of dosimeters calibration for radiotherapy in secondary dosimetric calibration laboratory of the CPHR; Automatizacion de la calibracion de dosimetros de radioterapia en el laboratorio secundario de calibracion dosimetrica del CPHR

    Energy Technology Data Exchange (ETDEWEB)

    Acosta, Andy L. Romero; Lores, Stefan Gutierrez, E-mail: c19btm@frcuba.co.cu [Centro de Proteccion e Higiene de las Radiaciones (CPHR), La Habana (Cuba)

    2013-11-01

    This paper presents the design and implementation of an automated system for measurements in the calibration of reference radiation dosimeters. It was made a software application that performs the acquisition of the measured values of electric charge, calculated calibration coefficient and automates the calibration certificate issuance. These values are stored in a log file on a PC. The use of the application improves control over the calibration process, helps to humanize the work and reduces personnel exposure. The tool developed has been applied to the calibration of dosimeters radiation patterns in the LSCD of the Centro de Proteccion e Higiene de las Radiaciones, Cuba.

  15. Automated calibration system for a high-precision measurement of neutrino mixing angle θ13 with the Daya Bay antineutrino detectors

    International Nuclear Information System (INIS)

    Liu, J.; Cai, B.; Carr, R.; Dwyer, D.A.; Gu, W.Q.; Li, G.S.; Qian, X.; McKeown, R.D.; Tsang, R.H.M.; Wang, W.; Wu, F.F.; Zhang, C.

    2014-01-01

    We describe the automated calibration system for the antineutrino detectors in the Daya Bay Neutrino Experiment. This system consists of 24 identical units instrumented on 8 identical 20-ton liquid scintillator detectors. Each unit is a fully automated robotic system capable of deploying an LED and various radioactive sources into the detector along given vertical axes. Selected results from performance studies of the calibration system are reported

  16. Automated calibration system for a high-precision measurement of neutrino mixing angle θ{sub 13} with the Daya Bay antineutrino detectors

    Energy Technology Data Exchange (ETDEWEB)

    Liu, J., E-mail: jianglai.liu@sjtu.edu.cn [Department of Physics, Shanghai Jiao Tong University, Shanghai (China); Kellogg Radiation Laboratory, California Institute of Technology, Pasadena, CA (United States); Cai, B.; Carr, R. [Kellogg Radiation Laboratory, California Institute of Technology, Pasadena, CA (United States); Dwyer, D.A. [Kellogg Radiation Laboratory, California Institute of Technology, Pasadena, CA (United States); Lawrence Berkeley National Laboratory, Berkeley, CA (United States); Gu, W.Q.; Li, G.S. [Department of Physics, Shanghai Jiao Tong University, Shanghai (China); Qian, X. [Kellogg Radiation Laboratory, California Institute of Technology, Pasadena, CA (United States); Brookhaven National Laboratory, Upton, NY (United States); McKeown, R.D. [Kellogg Radiation Laboratory, California Institute of Technology, Pasadena, CA (United States); Department of Physics, College of William and Mary, Williamsburg, VA (United States); Tsang, R.H.M. [Kellogg Radiation Laboratory, California Institute of Technology, Pasadena, CA (United States); Wang, W. [Department of Physics, College of William and Mary, Williamsburg, VA (United States); Wu, F.F. [Kellogg Radiation Laboratory, California Institute of Technology, Pasadena, CA (United States); Zhang, C. [Kellogg Radiation Laboratory, California Institute of Technology, Pasadena, CA (United States); Brookhaven National Laboratory, Upton, NY (United States)

    2014-06-01

    We describe the automated calibration system for the antineutrino detectors in the Daya Bay Neutrino Experiment. This system consists of 24 identical units instrumented on 8 identical 20-ton liquid scintillator detectors. Each unit is a fully automated robotic system capable of deploying an LED and various radioactive sources into the detector along given vertical axes. Selected results from performance studies of the calibration system are reported.

  17. Calibration and application of an automated seepage meter for monitoring water flow across the sediment-water interface.

    Science.gov (United States)

    Zhu, Tengyi; Fu, Dafang; Jenkinson, Byron; Jafvert, Chad T

    2015-04-01

    The advective flow of sediment pore water is an important parameter for understanding natural geochemical processes within lake, river, wetland, and marine sediments and also for properly designing permeable remedial sediment caps placed over contaminated sediments. Automated heat pulse seepage meters can be used to measure the vertical component of sediment pore water flow (i.e., vertical Darcy velocity); however, little information on meter calibration as a function of ambient water temperature exists in the literature. As a result, a method with associated equations for calibrating a heat pulse seepage meter as a function of ambient water temperature is fully described in this paper. Results of meter calibration over the temperature range 7.5 to 21.2 °C indicate that errors in accuracy are significant if proper temperature-dependence calibration is not performed. The proposed calibration method allows for temperature corrections to be made automatically in the field at any ambient water temperature. The significance of these corrections is discussed.

  18. A hardware-software system for the automation of verification and calibration of oil metering units secondary equipment

    Science.gov (United States)

    Boyarnikov, A. V.; Boyarnikova, L. V.; Kozhushko, A. A.; Sekachev, A. F.

    2017-08-01

    In the article the process of verification (calibration) of oil metering units secondary equipment is considered. The purpose of the work is to increase the reliability and reduce the complexity of this process by developing a software and hardware system that provides automated verification and calibration. The hardware part of this complex carries out the commutation of the measuring channels of the verified controller and the reference channels of the calibrator in accordance with the introduced algorithm. The developed software allows controlling the commutation of channels, setting values on the calibrator, reading the measured data from the controller, calculating errors and compiling protocols. This system can be used for checking the controllers of the secondary equipment of the oil metering units in the automatic verification mode (with the open communication protocol) or in the semi-automatic verification mode (without it). The peculiar feature of the approach used is the development of a universal signal switch operating under software control, which can be configured for various verification methods (calibration), which allows to cover the entire range of controllers of metering units secondary equipment. The use of automatic verification with the help of a hardware and software system allows to shorten the verification time by 5-10 times and to increase the reliability of measurements, excluding the influence of the human factor.

  19. Development of an automated scanning monochromator for sensitivity calibration of the MUSTANG instrument

    Science.gov (United States)

    Rivers, Thane D.

    1992-06-01

    An Automated Scanning Monochromator was developed using: an Acton Research Corporation (ARC) Monochromator, Ealing Photomultiplier Tube and a Macintosh PC in conjunction with LabVIEW software. The LabVIEW Virtual Instrument written to operate the ARC Monochromator is a mouse driven user friendly program developed for automated spectral data measurements. Resolution and sensitivity of the Automated Scanning Monochromator System were determined experimentally. The Automated monochromator was then used for spectral measurements of a Platinum Lamp. Additionally, the reflectivity curve for a BaSO4 coated screen has been measured. Reflectivity measurements indicate a large discrepancy with expected results. Further analysis of the reflectivity experiment is required for conclusive results.

  20. Camera calibration in a hazardous environment performed in situ with automated analysis and verification

    International Nuclear Information System (INIS)

    DePiero, F.W.; Kress, R.L.

    1993-01-01

    Camera calibration using the method of Two Planes is discussed. An implementation of the technique is described that may be performed in situ, e.g., in a hazardous or contaminated environment, thus eliminating the need for decontamination of camera systems before recalibration. Companion analysis techniques used for verifying the correctness of the calibration are presented

  1. Energy Modelling and Automated Calibrations of Ancient Building Simulations: A Case Study of a School in the Northwest of Spain

    Directory of Open Access Journals (Sweden)

    Ana Ogando

    2017-06-01

    Full Text Available In the present paper, the energy performance of buildings forming a school centre in the northwest of Spain was analyzed using a transient simulation of the energy model of the school, which was developed with TRNSYS, a software of proven reliability in the field of thermal simulations. A deterministic calibration approach was applied to the initial building model to adjust the predictions to the actual performance of the school, data acquired during the temperature measurement campaign. The buildings under study were in deteriorated conditions due to poor maintenance over the years, presenting a big challenge for modelling and simulating it in a reliable way. The results showed that the proposed methodology is successful for obtaining calibrated thermal models of these types of damaged buildings, as the metrics employed to verify the final error showed a reduced normalized mean bias error (NMBE of 2.73%. It was verified that a decrease of approximately 60% in NMBE and 17% in the coefficient of variation of the root mean square error (CV(RMSE was achieved due to the calibration process. Subsequent steps were performed with the aid of new software, which was developed under a European project that enabled the automated calibration of the simulations.

  2. Thrombin generation by activated factor VII on platelet activated by different agonists. Extending the cell-based model of hemostasis

    Directory of Open Access Journals (Sweden)

    Herrera Maria

    2006-04-01

    Full Text Available Abstract Background Platelet activation is crucial in normal hemostasis. Using a clotting system free of external tissue factor, we investigated whether activated Factor VII in combination with platelet agonists increased thrombin generation (TG in vitro. Methods and results TG was quantified by time parameters: lag time (LT and time to peak (TTP, and by amount of TG: peak of TG (PTG and area under thrombin formation curve after 35 minutes (AUC→35min in plasma from 29 healthy volunteers using the calibrated automated thrombography (CAT technique. TG parameters were measured at basal conditions and after platelet stimulation by sodium arachidonate (AA, ADP, and collagen (Col. In addition, the effects of recombinant activated FVII (rFVIIa alone or combined with the other platelet agonists on TG parameters were investigated. We found that LT and TTP were significantly decreased (p 35min were significantly increased (p 35min (but not PTG when compared to platelet rich plasma activated with agonists in the absence of rFVIIa. Conclusion Platelets activated by AA, ADP, Col or rFVIIa triggered TG. This effect was increased by combining rFVIIa with other agonists. Our intrinsic coagulation system produced a burst in TG independent of external tissue factor activity an apparent hemostatic effect with little thrombotic capacity. Thus we suggest a modification in the cell-based model of hemostasis.

  3. Plasma centrifugation does not influence thrombin-antithrombin and plasmin-antiplasmin levels but determines platelet microparticles count.

    Science.gov (United States)

    Stępień, Ewa; Gruszczyński, Krzysztof; Kapusta, Przemysław; Kowalik, Artur; Wybrańska, Iwona

    2015-01-01

    Centrifugation is an essential step for plasma preparation to remove residual elements in plasma, especially platelets and platelet-derived microparticles (PMPs). Our working hypothesis was that centrifugation as a preanalytical step may influence some coagulation parameters. Healthy young men were recruited (N=17). For centrifugation, two protocols were applied: (A) the first centrifugation at 2500xg for 15 min and (B) at 2500xg for 20 min at room temperature with a light brake. In protocol (A), the second centrifugation was carried out at 2500xg for 15 min, whereas in protocol (B), the second centrifugation involved a 10 min spin at 13,000 x g. Thrombin-antithrombin (TAT) and plasmin-antiplasmin (PAP) complexes concentrations were determined by enzyme-linked immunosorbent assays. PMPs were stained with CD41 antibody and annexin V, and analyzed by flow cytometry method. Procoagulant activity was assayed by the Calibrated Automated Thrombogram method as a slope of thrombin formation (CAT velocity). Median TAT and PAP concentrations did not differ between the centrifugation protocols. The high speed centrifugation reduced the median (IQR) PMP count in plasma from 1291 (841-1975) to 573 (391-1010) PMP/µL (P=0.001), and CAT velocity from 2.01 (1.31-2.88) to 0.97 (0.82-1.73) nM/min (P=0.049). Spearman's rank correlation analysis showed correlation between TAT and PMPs in the protocol A plasma which was (rho=0.52, PCentrifugation protocols do not influence the markers of plasminogen (PAP) and thrombin (TAT) generation but they do affect the PMP count and procoagulant activity.

  4. Modeling thrombin generation: plasma composition based approach.

    Science.gov (United States)

    Brummel-Ziedins, Kathleen E; Everse, Stephen J; Mann, Kenneth G; Orfeo, Thomas

    2014-01-01

    Thrombin has multiple functions in blood coagulation and its regulation is central to maintaining the balance between hemorrhage and thrombosis. Empirical and computational methods that capture thrombin generation can provide advancements to current clinical screening of the hemostatic balance at the level of the individual. In any individual, procoagulant and anticoagulant factor levels together act to generate a unique coagulation phenotype (net balance) that is reflective of the sum of its developmental, environmental, genetic, nutritional and pharmacological influences. Defining such thrombin phenotypes may provide a means to track disease progression pre-crisis. In this review we briefly describe thrombin function, methods for assessing thrombin dynamics as a phenotypic marker, computationally derived thrombin phenotypes versus determined clinical phenotypes, the boundaries of normal range thrombin generation using plasma composition based approaches and the feasibility of these approaches for predicting risk.

  5. Automated correction on X-rays calibration using transmission chamber and LabVIEWTM

    International Nuclear Information System (INIS)

    Betti, Flavio; Potiens, Maria da Penha Albuquerque

    2009-01-01

    Uncertainties during prolonged exposure times on X-rays calibration procedures at the Instruments Calibration facilities at IPEN may suffer from efficiency (and therefore intensity) variations on the industrial X-Ray generator used. Using a transmission chamber as an online reference chamber during the whole irradiation process is proposed in order to compensate for such error source. Also temperature (and pressure) fluctuations may arise from the performance limited calibration room air conditioning system. As an open ionization chamber, that monitor chamber does require calculation of a correction factor due to the temperature and pressure effects on air density. Sending and processing data from all related instruments (electrometer, thermometer and barometer) can be more easily achieved by interfacing them to a host computer running an especially developed algorithm using LabVIEW TM environment which will not only apply the proper correction factors during runtime, but also determine the exact length of time to reach a desired condition, which can be: time period, charge collected, or air kerma, based on the previous calibration of the whole system using a reference chamber traceable to primary standard dosimetry laboratories. When performing such calibration, two temperature sensors (secondary standard thermistors) are simultaneously used, one for the transmission chamber, and other for the reference chamber. As the substitution method is used during actual customer's calibration, the readings from the second thermistor can also be used when desired for further corrections. Use of LabVIEW TM programming language allowed for a shorter development time, and it is also extremely convenient to make things easier when improvements and modifications are called for. (author)

  6. Construction and calibration of a low cost and fully automated vibrating sample magnetometer

    Science.gov (United States)

    El-Alaily, T. M.; El-Nimr, M. K.; Saafan, S. A.; Kamel, M. M.; Meaz, T. M.; Assar, S. T.

    2015-07-01

    A low cost vibrating sample magnetometer (VSM) has been constructed by using an electromagnet and an audio loud speaker; where both are controlled by a data acquisition device. The constructed VSM records the magnetic hysteresis loop up to 8.3 KG at room temperature. The apparatus has been calibrated and tested by using magnetic hysteresis data of some ferrite samples measured by two scientifically calibrated magnetometers; model (Lake Shore 7410) and model (LDJ Electronics Inc. Troy, MI). Our VSM lab-built new design proved success and reliability.

  7. CONSTRUCTION OF A CALIBRATED PROBABILISTIC CLASSIFICATION CATALOG: APPLICATION TO 50k VARIABLE SOURCES IN THE ALL-SKY AUTOMATED SURVEY

    International Nuclear Information System (INIS)

    Richards, Joseph W.; Starr, Dan L.; Miller, Adam A.; Bloom, Joshua S.; Brink, Henrik; Crellin-Quick, Arien; Butler, Nathaniel R.

    2012-01-01

    With growing data volumes from synoptic surveys, astronomers necessarily must become more abstracted from the discovery and introspection processes. Given the scarcity of follow-up resources, there is a particularly sharp onus on the frameworks that replace these human roles to provide accurate and well-calibrated probabilistic classification catalogs. Such catalogs inform the subsequent follow-up, allowing consumers to optimize the selection of specific sources for further study and permitting rigorous treatment of classification purities and efficiencies for population studies. Here, we describe a process to produce a probabilistic classification catalog of variability with machine learning from a multi-epoch photometric survey. In addition to producing accurate classifications, we show how to estimate calibrated class probabilities and motivate the importance of probability calibration. We also introduce a methodology for feature-based anomaly detection, which allows discovery of objects in the survey that do not fit within the predefined class taxonomy. Finally, we apply these methods to sources observed by the All-Sky Automated Survey (ASAS), and release the Machine-learned ASAS Classification Catalog (MACC), a 28 class probabilistic classification catalog of 50,124 ASAS sources in the ASAS Catalog of Variable Stars. We estimate that MACC achieves a sub-20% classification error rate and demonstrate that the class posterior probabilities are reasonably calibrated. MACC classifications compare favorably to the classifications of several previous domain-specific ASAS papers and to the ASAS Catalog of Variable Stars, which had classified only 24% of those sources into one of 12 science classes.

  8. CONSTRUCTION OF A CALIBRATED PROBABILISTIC CLASSIFICATION CATALOG: APPLICATION TO 50k VARIABLE SOURCES IN THE ALL-SKY AUTOMATED SURVEY

    Energy Technology Data Exchange (ETDEWEB)

    Richards, Joseph W.; Starr, Dan L.; Miller, Adam A.; Bloom, Joshua S.; Brink, Henrik; Crellin-Quick, Arien [Astronomy Department, University of California, Berkeley, CA 94720-3411 (United States); Butler, Nathaniel R., E-mail: jwrichar@stat.berkeley.edu [School of Earth and Space Exploration, Arizona State University, Tempe, AZ 85287 (United States)

    2012-12-15

    With growing data volumes from synoptic surveys, astronomers necessarily must become more abstracted from the discovery and introspection processes. Given the scarcity of follow-up resources, there is a particularly sharp onus on the frameworks that replace these human roles to provide accurate and well-calibrated probabilistic classification catalogs. Such catalogs inform the subsequent follow-up, allowing consumers to optimize the selection of specific sources for further study and permitting rigorous treatment of classification purities and efficiencies for population studies. Here, we describe a process to produce a probabilistic classification catalog of variability with machine learning from a multi-epoch photometric survey. In addition to producing accurate classifications, we show how to estimate calibrated class probabilities and motivate the importance of probability calibration. We also introduce a methodology for feature-based anomaly detection, which allows discovery of objects in the survey that do not fit within the predefined class taxonomy. Finally, we apply these methods to sources observed by the All-Sky Automated Survey (ASAS), and release the Machine-learned ASAS Classification Catalog (MACC), a 28 class probabilistic classification catalog of 50,124 ASAS sources in the ASAS Catalog of Variable Stars. We estimate that MACC achieves a sub-20% classification error rate and demonstrate that the class posterior probabilities are reasonably calibrated. MACC classifications compare favorably to the classifications of several previous domain-specific ASAS papers and to the ASAS Catalog of Variable Stars, which had classified only 24% of those sources into one of 12 science classes.

  9. Efficient Photometry In-Frame Calibration (EPIC) Gaussian Corrections for Automated Background Normalization of Rate-Tracked Satellite Imagery

    Science.gov (United States)

    Griesbach, J.; Wetterer, C.; Sydney, P.; Gerber, J.

    Photometric processing of non-resolved Electro-Optical (EO) images has commonly required the use of dark and flat calibration frames that are obtained to correct for charge coupled device (CCD) dark (thermal) noise and CCD quantum efficiency/optical path vignetting effects respectively. It is necessary to account/calibrate for these effects so that the brightness of objects of interest (e.g. stars or resident space objects (RSOs)) may be measured in a consistent manner across the CCD field of view. Detected objects typically require further calibration using aperture photometry to compensate for sky background (shot noise). For this, annuluses are measured around each detected object whose contained pixels are used to estimate an average background level that is subtracted from the detected pixel measurements. In a new photometric calibration software tool developed for AFRL/RD, called Efficient Photometry In-Frame Calibration (EPIC), an automated background normalization technique is proposed that eliminates the requirement to capture dark and flat calibration images. The proposed technique simultaneously corrects for dark noise, shot noise, and CCD quantum efficiency/optical path vignetting effects. With this, a constant detection threshold may be applied for constant false alarm rate (CFAR) object detection without the need for aperture photometry corrections. The detected pixels may be simply summed (without further correction) for an accurate instrumental magnitude estimate. The noise distribution associated with each pixel is assumed to be sampled from a Poisson distribution. Since Poisson distributed data closely resembles Gaussian data for parameterized means greater than 10, the data may be corrected by applying bias subtraction and standard-deviation division. EPIC performs automated background normalization on rate-tracked satellite images using the following technique. A deck of approximately 50-100 images is combined by performing an independent median

  10. Increased thrombin generation in a mouse model of cancer cachexia is partially interleukin-6 dependent.

    Science.gov (United States)

    Reddel, C J; Allen, J D; Ehteda, A; Taylor, R; Chen, V M Y; Curnow, J L; Kritharides, L; Robertson, G

    2017-03-01

    Essentials Cancer cachexia and cancer-associated thrombosis have not previously been mechanistically linked. We assessed thrombin generation and coagulation parameters in cachectic C26 tumor-bearing mice. C26 mice are hypercoagulable, partially corrected by blocking tumor derived interleukin-6. Coagulability and anti-inflammatory interventions may be clinically important in cancer cachexia. Background Cancer cachexia and cancer-associated thrombosis are potentially fatal outcomes of advanced cancer, which have not previously been mechanistically linked. The colon 26 (C26) carcinoma is a well-established mouse model of complications of advanced cancer cachexia, partially dependent on high levels of interleukin-6 (IL-6) produced by the tumor. Objectives To assess if cancer cachexia altered the coagulation state and if this was attributable to tumor IL-6 production. Methods In male BALB/c*DBA2 (F1 hybrid) mice with a C26 tumor we used modified calibrated automated thrombogram and fibrin generation (based on overall hemostatic potential) assays to assess the functional coagulation state, and also examined fibrinogen, erythrocyte sedimentation rate (ESR), platelet count, tissue factor pathway inhibitor (TFPI) and hepatic expression of coagulation factors by microarray. C26 mice were compared with non-cachectic NC26, pair-fed and sham control mice. IL-6 expression in C26 cells was knocked down by lentiviral shRNA constructs. Results C26 mice with significant weight loss and highly elevated IL-6 had elevated thrombin generation, fibrinogen, ESR, platelets and TFPI compared with all control groups. Fibrin generation was elevated compared with pair-fed and sham controls but not compared with NC26 tumor mice. Hepatic expression of coagulation factors and fibrinolytic inhibitors was increased. Silencing IL-6 in the tumor significantly, but incompletely, attenuated the increased thrombin generation, fibrinogen and TFPI. Conclusions Cachectic C26 tumor-bearing mice are in a

  11. RADIOLOGICAL TIPS Percutaneous thrombin injection for ...

    African Journals Online (AJOL)

    pseudoaneurysm sac has a typical 'yin-yang' sign. The neck is normally seen posteriorly and is usually thin and longitudinal. A large neck diameter (e.g. >10 mm) is a relative contra-indication for thrombin injection because of a slightly higher risk of distal embolisation. There are several thrombin preparations available.

  12. Aptamer Based Microsphere Biosensor for Thrombin Detection

    Directory of Open Access Journals (Sweden)

    Xudong Fan

    2006-08-01

    Full Text Available We have developed an optical microsphere resonator biosensor using aptamer asreceptor for the measurement of the important biomolecule thrombin. The sphere surface ismodified with anti-thrombin aptamer, which has excellent binding affinity and selectivityfor thrombin. Binding of the thrombin at the sphere surface is monitored by the spectralposition of the microsphere’s whispering gallery mode resonances. A detection limit on theorder of 1 NIH Unit/mL is demonstrated. Control experiments with non-aptameroligonucleotide and BSA are also carried out to confirm the specific binding betweenaptamer and thrombin. We expect that this demonstration will lead to the development ofhighly sensitive biomarker sensors based on aptamer with lower cost and higher throughputthan current technology.

  13. Automated System Calibration and Verification of the Position Measurements for the Los Alamos Isotope Production Facility and the Switchyard Kicker Facilities

    Science.gov (United States)

    Barr, D.; Gilpatrick, J. D.; Martinez, D.; Shurter, R. B.

    2004-11-01

    The Los Alamos Neutron Science Center (LANSCE) facility at Los Alamos National Laboratory has constructed both an Isotope Production Facility (IPF) and a Switchyard Kicker (XDK) as additions to the H+ and H- accelerator. These additions contain eleven Beam Position Monitors (BPMs) that measure the beam's position throughout the transport. The analog electronics within each processing module determines the beam position using the log-ratio technique. For system reliability, calibrations compensate for various temperature drifts and other imperfections in the processing electronics components. Additionally, verifications are periodically implemented by a PC running a National Instruments LabVIEW virtual instrument (VI) to verify continued system and cable integrity. The VI communicates with the processor cards via a PCI/MXI-3 VXI-crate communication module. Previously, accelerator operators performed BPM system calibrations typically once per day while beam was explicitly turned off. One of this new measurement system's unique achievements is its automated calibration and verification capability. Taking advantage of the pulsed nature of the LANSCE-facility beams, the integrated electronics hardware and VI perform calibration and verification operations between beam pulses without interrupting production beam delivery. The design, construction, and performance results of the automated calibration and verification portion of this position measurement system will be the topic of this paper.

  14. Extracellular Histones Increase Tissue Factor Activity and Enhance Thrombin Generation by Human Blood Monocytes.

    Science.gov (United States)

    Gould, Travis J; Lysov, Zakhar; Swystun, Laura L; Dwivedi, Dhruva J; Zarychanski, Ryan; Fox-Robichaud, Alison E; Liaw, Patricia C

    2016-12-01

    Sepsis is characterized by systemic activation of inflammatory and coagulation pathways in response to infection. Recently, it was demonstrated that histones released into the circulation by dying/activated cells may contribute to sepsis pathology. Although the ability of extracellular histones to modulate the procoagulant activities of several cell types has been investigated, the influence of histones on the hemostatic functions of circulating monocytes is unknown. To address this, we investigated the ability of histones to modulate the procoagulant potential of THP-1 cells and peripheral blood monocytes, and examined the effects of plasmas obtained from septic patients to induce a procoagulant phenotype on monocytic cells. Tissue factor (TF) activity assays were performed on histone-treated THP-1 cells and blood monocytes. Exposure of monocytic cells to histones resulted in increases in TF activity, TF antigen, and phosphatidylserine exposure. Histones modulate the procoagulant activity via engagement of Toll-like receptors 2 and 4, and this effect was abrogated with inhibitory antibodies. Increased TF activity of histone-treated cells corresponded to enhanced thrombin generation in plasma determined by calibrated automated thrombography. Finally, TF activity was increased on monocytes exposed to plasma from septic patients, an effect that was attenuated in plasma from patients receiving unfractionated heparin (UFH). Our studies suggest that increased levels of extracellular histones found in sepsis contribute to dysregulated coagulation by increasing TF activity of monocytes. These procoagulant effects can be partially ameliorated in sepsis patients receiving UFH, thereby identifying extracellular histones as a potential therapeutic target for sepsis treatment.

  15. 3D camera assisted fully automated calibration of scanning laser Doppler vibrometers

    International Nuclear Information System (INIS)

    Sels, Seppe; Ribbens, Bart; Mertens, Luc; Vanlanduit, Steve

    2016-01-01

    Scanning laser Doppler vibrometers (LDV) are used to measure full-field vibration shapes of products and structures. In most commercially available scanning laser Doppler vibrometer systems the user manually draws a grid of measurement locations on a 2D camera image of the product. The determination of the correct physical measurement locations can be a time consuming and diffcult task. In this paper we present a new methodology for product testing and quality control that integrates 3D imaging techniques with vibration measurements. This procedure allows to test prototypes in a shorter period because physical measurements locations will be located automatically. The proposed methodology uses a 3D time-of-flight camera to measure the location and orientation of the test-object. The 3D image of the time-of-flight camera is then matched with the 3D-CAD model of the object in which measurement locations are pre-defined. A time of flight camera operates strictly in the near infrared spectrum. To improve the signal to noise ratio in the time-of-flight measurement, a time-of-flight camera uses a band filter. As a result of this filter, the laser spot of most laser vibrometers is invisible in the time-of-flight image. Therefore a 2D RGB-camera is used to find the laser-spot of the vibrometer. The laser spot is matched to the 3D image obtained by the time-of-flight camera. Next an automatic calibration procedure is used to aim the laser at the (pre)defined locations. Another benefit from this methodology is that it incorporates automatic mapping between a CAD model and the vibration measurements. This mapping can be used to visualize measurements directly on a 3D CAD model. Secondly the orientation of the CAD model is known with respect to the laser beam. This information can be used to find the direction of the measured vibration relatively to the surface of the object. With this direction, the vibration measurements can be compared more precisely with numerical

  16. 3D camera assisted fully automated calibration of scanning laser Doppler vibrometers

    Energy Technology Data Exchange (ETDEWEB)

    Sels, Seppe, E-mail: Seppe.Sels@uantwerpen.be; Ribbens, Bart; Mertens, Luc; Vanlanduit, Steve [Op3Mech Research Group, University of Antwerp, Salesianenlaan 90, 2660 Antwerp (Belgium)

    2016-06-28

    Scanning laser Doppler vibrometers (LDV) are used to measure full-field vibration shapes of products and structures. In most commercially available scanning laser Doppler vibrometer systems the user manually draws a grid of measurement locations on a 2D camera image of the product. The determination of the correct physical measurement locations can be a time consuming and diffcult task. In this paper we present a new methodology for product testing and quality control that integrates 3D imaging techniques with vibration measurements. This procedure allows to test prototypes in a shorter period because physical measurements locations will be located automatically. The proposed methodology uses a 3D time-of-flight camera to measure the location and orientation of the test-object. The 3D image of the time-of-flight camera is then matched with the 3D-CAD model of the object in which measurement locations are pre-defined. A time of flight camera operates strictly in the near infrared spectrum. To improve the signal to noise ratio in the time-of-flight measurement, a time-of-flight camera uses a band filter. As a result of this filter, the laser spot of most laser vibrometers is invisible in the time-of-flight image. Therefore a 2D RGB-camera is used to find the laser-spot of the vibrometer. The laser spot is matched to the 3D image obtained by the time-of-flight camera. Next an automatic calibration procedure is used to aim the laser at the (pre)defined locations. Another benefit from this methodology is that it incorporates automatic mapping between a CAD model and the vibration measurements. This mapping can be used to visualize measurements directly on a 3D CAD model. Secondly the orientation of the CAD model is known with respect to the laser beam. This information can be used to find the direction of the measured vibration relatively to the surface of the object. With this direction, the vibration measurements can be compared more precisely with numerical

  17. Automated characterization of perceptual quality of clinical chest radiographs: Validation and calibration to observer preference

    International Nuclear Information System (INIS)

    Samei, Ehsan; Lin, Yuan; Choudhury, Kingshuk R.; Page McAdams, H.

    2014-01-01

    Purpose: The authors previously proposed an image-based technique [Y. Lin et al. Med. Phys. 39, 7019–7031 (2012)] to assess the perceptual quality of clinical chest radiographs. In this study, an observer study was designed and conducted to validate the output of the program against rankings by expert radiologists and to establish the ranges of the output values that reflect the acceptable image appearance so the program output can be used for image quality optimization and tracking. Methods: Using an IRB-approved protocol, 2500 clinical chest radiographs (PA/AP) were collected from our clinical operation. The images were processed through our perceptual quality assessment program to measure their appearance in terms of ten metrics of perceptual image quality: lung gray level, lung detail, lung noise, rib–lung contrast, rib sharpness, mediastinum detail, mediastinum noise, mediastinum alignment, subdiaphragm–lung contrast, and subdiaphragm area. From the results, for each targeted appearance attribute/metric, 18 images were selected such that the images presented a relatively constant appearance with respect to all metrics except the targeted one. The images were then incorporated into a graphical user interface, which displayed them into three panels of six in a random order. Using a DICOM calibrated diagnostic display workstation and under low ambient lighting conditions, each of five participating attending chest radiologists was tasked to spatially order the images based only on the targeted appearance attribute regardless of the other qualities. Once ordered, the observer also indicated the range of image appearances that he/she considered clinically acceptable. The observer data were analyzed in terms of the correlations between the observer and algorithmic rankings and interobserver variability. An observer-averaged acceptable image appearance was also statistically derived for each quality attribute based on the collected individual acceptable ranges

  18. Automated characterization of perceptual quality of clinical chest radiographs: Validation and calibration to observer preference

    Energy Technology Data Exchange (ETDEWEB)

    Samei, Ehsan, E-mail: samei@duke.edu [Carl E. Ravin Advanced Imaging Laboratories, Departments of Radiology, Physics, Biomedical Engineering, Electrical and Computer Engineering, Medical Physics Graduate Program, Duke Clinical Imaging Physics Group, Duke University, Durham, North Carolina 27710 (United States); Lin, Yuan [Carl E. Ravin Advanced Imaging Laboratories, Departments of Radiology and Physics, Duke University, Durham, North Carolina 27710 (United States); Choudhury, Kingshuk R. [Carl E. Ravin Advanced Imaging Laboratories, Department of Radiology and Biostatistics and Bioinformatics, Duke University, Durham, North Carolina 27710 (United States); Page McAdams, H. [Carl E. Ravin Advanced Imaging Laboratories, Department of Radiology, Duke University, Durham, North Carolina 27710 (United States)

    2014-11-01

    Purpose: The authors previously proposed an image-based technique [Y. Lin et al. Med. Phys. 39, 7019–7031 (2012)] to assess the perceptual quality of clinical chest radiographs. In this study, an observer study was designed and conducted to validate the output of the program against rankings by expert radiologists and to establish the ranges of the output values that reflect the acceptable image appearance so the program output can be used for image quality optimization and tracking. Methods: Using an IRB-approved protocol, 2500 clinical chest radiographs (PA/AP) were collected from our clinical operation. The images were processed through our perceptual quality assessment program to measure their appearance in terms of ten metrics of perceptual image quality: lung gray level, lung detail, lung noise, rib–lung contrast, rib sharpness, mediastinum detail, mediastinum noise, mediastinum alignment, subdiaphragm–lung contrast, and subdiaphragm area. From the results, for each targeted appearance attribute/metric, 18 images were selected such that the images presented a relatively constant appearance with respect to all metrics except the targeted one. The images were then incorporated into a graphical user interface, which displayed them into three panels of six in a random order. Using a DICOM calibrated diagnostic display workstation and under low ambient lighting conditions, each of five participating attending chest radiologists was tasked to spatially order the images based only on the targeted appearance attribute regardless of the other qualities. Once ordered, the observer also indicated the range of image appearances that he/she considered clinically acceptable. The observer data were analyzed in terms of the correlations between the observer and algorithmic rankings and interobserver variability. An observer-averaged acceptable image appearance was also statistically derived for each quality attribute based on the collected individual acceptable ranges

  19. Contributions of procoagulants and anticoagulants to the international normalized ratio and thrombin generation assay in patients treated with warfarin: potential role of protein Z as a powerful determinant of coagulation assays.

    Science.gov (United States)

    Choi, Qute; Kim, Ji-Eun; Hyun, Jungwon; Han, Kyou-Sup; Kim, Hyun Kyung

    2013-07-01

    The effects of warfarin are measured with the international normalized ratio (INR). However, the thrombin generation assay (TGA) may offer more information about global coagulation. We analyzed the monitoring performance of the TGA and INR and investigated the impact of procoagulants (fibrinogen, factor (F)II, FVII, FIX, and FX) and anticoagulants (proteins C, S, and Z) on them. The TGA was performed on a calibrated automated thrombogram, producing lag time, endogenous thrombin potential (ETP), and peak thrombin in 239 patients treated with warfarin. Pro- and anticoagulant levels were also measured. The INR was significantly and inversely correlated with ETP. The therapeutic range of ETP comparable to an INR range of 2.0-3.0 was 290.1-494.6. ETP showed comparable performance to the INR as a warfarin-monitoring parameter with respect to clinical complication rate. The median levels of FII, FVII, FIX, and FX and proteins C and Z tended to decrease gradually with increasing anticoagulation intensity according to the INR or ETP. Of note, protein Z levels decreased dramatically with increasing anticoagulation status. INRs were significantly determined by FII, FVII, and protein Z. ETP was significantly dependent on FVII, and proteins C and Z concentration. Protein Z significantly reduced the total amount of thrombin generation and prolonged PT value in vitro. The INR and ETP exhibit similar efficacy for warfarin monitoring according to the clinical complication rate. Protein Z is considered to be a significant determinant of INR and ETP in patients on warfarin therapy. Copyright © 2013 Elsevier Ltd. All rights reserved.

  20. Automated calibration of laser spectrometer measurements of δ18 O and δ2 H values in water vapour using a Dew Point Generator.

    Science.gov (United States)

    Munksgaard, Niels C; Cheesman, Alexander W; Gray-Spence, Andrew; Cernusak, Lucas A; Bird, Michael I

    2018-06-30

    Continuous measurement of stable O and H isotope compositions in water vapour requires automated calibration for remote field deployments. We developed a new low-cost device for calibration of both water vapour mole fraction and isotope composition. We coupled a commercially available dew point generator (DPG) to a laser spectrometer and developed hardware for water and air handling along with software for automated operation and data processing. We characterised isotopic fractionation in the DPG, conducted a field test and assessed the influence of critical parameters on the performance of the device. An analysis time of 1 hour was sufficient to achieve memory-free analysis of two water vapour standards and the δ 18 O and δ 2 H values were found to be independent of water vapour concentration over a range of ≈20,000-33,000 ppm. The reproducibility of the standard vapours over a 10-day period was better than 0.14 ‰ and 0.75 ‰ for δ 18 O and δ 2 H values, respectively (1 σ, n = 11) prior to drift correction and calibration. The analytical accuracy was confirmed by the analysis of a third independent vapour standard. The DPG distillation process requires that isotope calibration takes account of DPG temperature, analysis time, injected water volume and air flow rate. The automated calibration system provides high accuracy and precision and is a robust, cost-effective option for long-term field measurements of water vapour isotopes. The necessary modifications to the DPG are minor and easily reversible. Copyright © 2018 John Wiley & Sons, Ltd.

  1. A near-infrared fluorescent bioassay for thrombin using aptamer-modified CuInS2 quantum dots

    International Nuclear Information System (INIS)

    Lin, Zihan; Hu, Tianyu; Liu, Ziping; Su, Xingguang; Pan, Dong

    2015-01-01

    We describe a near-infrared (NIR) fluorescent thrombin assay using a thrombin-binding aptamer (TBA) and Zn(II)-activated CuInS 2 quantum dots (Q-dots). The fluorescence of Zn(II)-activated Q-dots is quenched by the TBA via photoinduced electron transfer, but if thrombin is added, it will bind to TBA to form G-quadruplexes and the Q-dots are released. As a result, the fluorescence intensity of the system is restored. This effect was exploited to design an assay for thrombin whose calibration plot, under optimum conditions, is linear in the 0.034 to 102 nmol L −1 concentration range, with a 12 pmol L −1 detection limit. The method is fairly simple, fast, and due to its picomolar detection limits holds great potential in the diagnosis of diseases associated with coagulation abnormalities and certain kinds of cancer. (author)

  2. Effects of recombinant human prothrombin on thrombin generation in plasma from patients with hemophilia A and B.

    Science.gov (United States)

    Hansson, K M; Gustafsson, D; Skärby, T; Frison, L; Berntorp, E

    2015-07-01

    The present study was carried out to investigate the impact of FII levels, and their increase, on the hemostatic potential in plasma from hemophilia A and B patients with and without inhibitors. Recombinant human factor (F) II (rhFII) was added ex vivo to plasma from 68 patients with hemophilia A and B, with or without inhibitors. The hemostatic potential as measured by thrombin generation (calibrated automated thrombogram [CAT]) was focused on the endogenous thrombin potential (ETP) as it has been shown to correlate with the clinical phenotype of bleeding in hemophilia patients and has also been used to guide bypassing therapy in hemophilia patients with inhibitors before elective surgery. The factor eight inhibitor bypassing agent (FEIBA(®) ) was used as a reference to the clinical situation. The study shows that rhFII concentration-dependently increased ETP by a similar magnitude in hemophilia A and B, both with and without inhibitors. Compared with FEIBA, rhFII showed a shallower concentration-response curve. In both types of hemophilia 100 mg L(-1) of rhFII roughly doubled the ETP. A corresponding response was obtained by 0.5 U mL(-1) of FEIBA. These data support the theory that FII is one of the major components responsible for the efficacy of FEIBA. The data also indicate that rhFII may be useful, alone or in combination with other coagulation factors, in some of the conditions for which FEIBA is used today, although more data are needed to substantiate this. © 2015 International Society on Thrombosis and Haemostasis.

  3. A New Colorimetrically-Calibrated Automated Video-Imaging Protocol for Day-Night Fish Counting at the OBSEA Coastal Cabled Observatory

    Directory of Open Access Journals (Sweden)

    Joaquín del Río

    2013-10-01

    Full Text Available Field measurements of the swimming activity rhythms of fishes are scant due to the difficulty of counting individuals at a high frequency over a long period of time. Cabled observatory video monitoring allows such a sampling at a high frequency over unlimited periods of time. Unfortunately, automation for the extraction of biological information (i.e., animals’ visual counts per unit of time is still a major bottleneck. In this study, we describe a new automated video-imaging protocol for the 24-h continuous counting of fishes in colorimetrically calibrated time-lapse photographic outputs, taken by a shallow water (20 m depth cabled video-platform, the OBSEA. The spectral reflectance value for each patch was measured between 400 to 700 nm and then converted into standard RGB, used as a reference for all subsequent calibrations. All the images were acquired within a standardized Region Of Interest (ROI, represented by a 2 × 2 m methacrylate panel, endowed with a 9-colour calibration chart, and calibrated using the recently implemented “3D Thin-Plate Spline” warping approach in order to numerically define color by its coordinates in n-dimensional space. That operation was repeated on a subset of images, 500 images as a training set, manually selected since acquired under optimum visibility conditions. All images plus those for the training set were ordered together through Principal Component Analysis allowing the selection of 614 images (67.6% out of 908 as a total corresponding to 18 days (at 30 min frequency. The Roberts operator (used in image processing and computer vision for edge detection was used to highlights regions of high spatial colour gradient corresponding to fishes’ bodies. Time series in manual and visual counts were compared together for efficiency evaluation. Periodogram and waveform analysis outputs provided very similar results, although quantified parameters in relation to the strength of respective rhythms were

  4. A new colorimetrically-calibrated automated video-imaging protocol for day-night fish counting at the OBSEA coastal cabled observatory.

    Science.gov (United States)

    del Río, Joaquín; Aguzzi, Jacopo; Costa, Corrado; Menesatti, Paolo; Sbragaglia, Valerio; Nogueras, Marc; Sarda, Francesc; Manuèl, Antoni

    2013-10-30

    Field measurements of the swimming activity rhythms of fishes are scant due to the difficulty of counting individuals at a high frequency over a long period of time. Cabled observatory video monitoring allows such a sampling at a high frequency over unlimited periods of time. Unfortunately, automation for the extraction of biological information (i.e., animals' visual counts per unit of time) is still a major bottleneck. In this study, we describe a new automated video-imaging protocol for the 24-h continuous counting of fishes in colorimetrically calibrated time-lapse photographic outputs, taken by a shallow water (20 m depth) cabled video-platform, the OBSEA. The spectral reflectance value for each patch was measured between 400 to 700 nm and then converted into standard RGB, used as a reference for all subsequent calibrations. All the images were acquired within a standardized Region Of Interest (ROI), represented by a 2 × 2 m methacrylate panel, endowed with a 9-colour calibration chart, and calibrated using the recently implemented "3D Thin-Plate Spline" warping approach in order to numerically define color by its coordinates in n-dimensional space. That operation was repeated on a subset of images, 500 images as a training set, manually selected since acquired under optimum visibility conditions. All images plus those for the training set were ordered together through Principal Component Analysis allowing the selection of 614 images (67.6%) out of 908 as a total corresponding to 18 days (at 30 min frequency). The Roberts operator (used in image processing and computer vision for edge detection) was used to highlights regions of high spatial colour gradient corresponding to fishes' bodies. Time series in manual and visual counts were compared together for efficiency evaluation. Periodogram and waveform analysis outputs provided very similar results, although quantified parameters in relation to the strength of respective rhythms were different. Results

  5. Topical thrombin-related corneal calcification.

    Science.gov (United States)

    Kiratli, Hayyam; Irkeç, Murat; Alaçal, Sibel; Söylemezoğlu, Figen

    2006-09-01

    To report a highly unusual case of corneal calcification after brief intraoperative use of topical thrombin. A 44-year-old man underwent sclerouvectomy for ciliochoroidal leiomyoma, during which 35 UNIH/mL lyophilized bovine thrombin mixed with 9 mL of diluent containing 1500 mmol/mL calcium chloride was used. From the first postoperative day, corneal and anterior lenticular capsule calcifications developed, and corneal involvement slightly enlarged thereafter. A year later, 2 corneal punch biopsies confirmed calcification mainly in the Bowman layer. Topical treatment with 1.5% ethylenediaminetetraacetic acid significantly restored corneal clarity. Six months later, a standard extracapsular cataract extraction with intraocular lens placement improved visual acuity to 20/60. This case suggests that topical thrombin drops with elevated calcium concentrations may cause acute corneal calcification in Bowman layer and on the anterior lens capsule.

  6. Use of a robotic manipulator in the simulation of the automation of a calibration process of dosemeters

    International Nuclear Information System (INIS)

    Benitez R, J.S.; Najera H, M.C.

    2002-01-01

    The development of a system based in a manipulative robot which simulates the operative sequence in a calibration process of dosemeters is presented. In this process it is performed the monitoring of the dosemeter positions and the calibrator by mean of an arm of articulated robot which develops the movement sequences and the taking a decision based on the information coming from the external sensors. (Author)

  7. Red blood cell-derived microparticles isolated from blood units initiate and propagate thrombin generation.

    Science.gov (United States)

    Rubin, Olivier; Delobel, Julien; Prudent, Michel; Lion, Niels; Kohl, Kid; Tucker, Erik I; Tissot, Jean-Daniel; Angelillo-Scherrer, Anne

    2013-08-01

    Red blood cell-derived microparticles (RMPs) are small phospholipid vesicles shed from RBCs in blood units, where they accumulate during storage. Because microparticles are bioactive, it could be suggested that RMPs are mediators of posttransfusion complications or, on the contrary, constitute a potential hemostatic agent. This study was performed to establish the impact on coagulation of RMPs isolated from blood units. Using calibrated automated thrombography, we investigated whether RMPs affect thrombin generation (TG) in plasma. We found that RMPs were not only able to increase TG in plasma in the presence of a low exogenous tissue factor (TF) concentration, but also to initiate TG in plasma in absence of exogenous TF. TG induced by RMPs in the absence of exogenous TF was neither affected by the presence of blocking anti-TF nor by the absence of Factor (F)VII. It was significantly reduced in plasma deficient in FVIII or F IX and abolished in FII-, FV-, FX-, or FXI-deficient plasma. TG was also totally abolished when anti-XI 01A6 was added in the sample. Finally, neither Western blotting, flow cytometry, nor immunogold labeling allowed the detection of traces of TF antigen. In addition, RMPs did not comprise polyphosphate, an important modulator of coagulation. Taken together, our data show that RMPs have FXI-dependent procoagulant properties and are able to initiate and propagate TG. The anionic surface of RMPs might be the site of FXI-mediated TG amplification and intrinsic tenase and prothrombinase complex assembly. © 2012 American Association of Blood Banks.

  8. Automating calibration, sensitivity and uncertainty analysis of complex models using the R package Flexible Modeling Environment (FME): SWAT as an example

    Science.gov (United States)

    Wu, Y.; Liu, S.

    2012-01-01

    Parameter optimization and uncertainty issues are a great challenge for the application of large environmental models like the Soil and Water Assessment Tool (SWAT), which is a physically-based hydrological model for simulating water and nutrient cycles at the watershed scale. In this study, we present a comprehensive modeling environment for SWAT, including automated calibration, and sensitivity and uncertainty analysis capabilities through integration with the R package Flexible Modeling Environment (FME). To address challenges (e.g., calling the model in R and transferring variables between Fortran and R) in developing such a two-language coupling framework, 1) we converted the Fortran-based SWAT model to an R function (R-SWAT) using the RFortran platform, and alternatively 2) we compiled SWAT as a Dynamic Link Library (DLL). We then wrapped SWAT (via R-SWAT) with FME to perform complex applications including parameter identifiability, inverse modeling, and sensitivity and uncertainty analysis in the R environment. The final R-SWAT-FME framework has the following key functionalities: automatic initialization of R, running Fortran-based SWAT and R commands in parallel, transferring parameters and model output between SWAT and R, and inverse modeling with visualization. To examine this framework and demonstrate how it works, a case study simulating streamflow in the Cedar River Basin in Iowa in the United Sates was used, and we compared it with the built-in auto-calibration tool of SWAT in parameter optimization. Results indicate that both methods performed well and similarly in searching a set of optimal parameters. Nonetheless, the R-SWAT-FME is more attractive due to its instant visualization, and potential to take advantage of other R packages (e.g., inverse modeling and statistical graphics). The methods presented in the paper are readily adaptable to other model applications that require capability for automated calibration, and sensitivity and uncertainty

  9. Retro-binding thrombin active site inhibitors: identification of an orally active inhibitor of thrombin catalytic activity.

    Science.gov (United States)

    Iwanowicz, Edwin J; Kimball, S David; Lin, James; Lau, Wan; Han, W-C; Wang, Tammy C; Roberts, Daniel G M; Schumacher, W A; Ogletree, Martin L; Seiler, Steven M

    2002-11-04

    A series of retro-binding inhibitors of human alpha-thrombin was prepared to elucidate structure-activity relationships (SAR) and optimize in vivo performance. Compounds 9 and 11, orally active inhibitors of thrombin catalytic activity, were identified to be efficacious in a thrombin-induced lethality model in mice.

  10. Development and implementation of an automated system for antiquated of the process of gamma radiation monitors calibration

    International Nuclear Information System (INIS)

    Silva Junior, Iremar Alves

    2012-01-01

    In this study it was carried out the development and implementation of a system for the appropriate process of gamma radiation monitors calibration, constituted by a pneumatic dispositive to exchange the attenuators and a positioning table, both actuated through a control panel. We also implemented a System of Caesa-Gammatron Irradiator, which increased the range of the air kerma rates, due to its higher activity comparing with the current system of gamma radiation in use in the calibration laboratory of gamma irradiation. Hence, it was necessary the installation of an attenuator dispositive remotely controlled in this irradiator system. Lastly, it was carried out an evaluation of the reduction in the rates of the occupational dose. This dissertation was developed with the aim of improving the quality of the services of calibration and tests of gamma radiation monitors - provided by the IPEN Laboratory of Instrument Calibration - as well as decreasing the occupational dose of the technicians involved in the process of calibration, following thus the principles of radiation protection. (author)

  11. Endogenous thrombin potential in polycystic ovary syndrome

    DEFF Research Database (Denmark)

    Aziz, Mubeena; Sidelmann, Johannes Jakobsen; Wissing, Marie Louise Muff

    2015-01-01

    OBJECTIVES: The objective of this study is to investigate plasma endogenous thrombin generation in four different phenotypes of polycystic ovary syndrome (PCOS) defined by Body Mass Index (BMI) and insulin resistance (IR). PCOS is diagnosed according to the Rotterdam criteria. DESIGN: Multicenter...

  12. Discovery of thrombin activatable fibrinolysis inhibitor (TAFI)

    NARCIS (Netherlands)

    Bertina, R.M.; Tilburg, N.H. van; Haverkate, F.; Bouma, B.N.; Borne, P.A.K. von dem; Meijers, J.C.M.; Campbell, W.; Eaton, D.; Hendriks, D.F.; Willemse, J.L.

    2006-01-01

    CAS: blood clotting factor 11, 9013-55-2; thrombin, 9002-04-4; tissue plasminogen activator, 105913-11-9; protein C, 60202-16-6; Carboxypeptidase U, 3.4.17.20; Protein C; Tissue Plasminogen Activator, 3.4.21.68

  13. Decreased prothrombin conversion and reduced thrombin inactivation explain rebalanced thrombin generation in liver cirrhosis.

    Directory of Open Access Journals (Sweden)

    Romy M W Kremers

    Full Text Available Impaired coagulation factor synthesis in cirrhosis causes a reduction of most pro- and anticoagulant factors. Cirrhosis patients show no clear bleeding or thrombotic phenotype, although they are at risk for both types of hemostatic event. Thrombin generation (TG is a global coagulation test and its outcome depends on underlying pro- and anticoagulant processes (prothrombin conversion and thrombin inactivation. We quantified the prothrombin conversion and thrombin inactivation during TG in 30 healthy subjects and 52 Child-Pugh (CP- A, 15 CP-B and 6 CP-C cirrhosis patients to test the hypothesis that coagulation is rebalanced in liver cirrhosis patients. Both prothrombin conversion and thrombin inactivation are reduced in cirrhosis patients. The effect on pro- and anticoagulant processes partially cancel each other out and as a result TG is comparable at 5 pM tissue factor between healthy subjects and patients. This supports the hypothesis of rebalanced hemostasis, as TG in cirrhosis patients remains within the normal range, despite large changes in prothrombin conversion and thrombin inactivation. Nevertheless, in silico analysis shows that normalization of either prothrombin conversion or thrombin inactivation to physiological levels, by for example the administration of prothrombin complex concentrates would cause an elevation of TG, whereas the normalization of both simultaneously maintains a balanced TG. Therefore, cirrhosis patients might require adapted hemostatic treatment.

  14. Automated air-void system characterization of hardened concrete: Helping computers to count air-voids like people count air-voids---Methods for flatbed scanner calibration

    Science.gov (United States)

    Peterson, Karl

    Since the discovery in the late 1930s that air entrainment can improve the durability of concrete, it has been important for people to know the quantity, spacial distribution, and size distribution of the air-voids in their concrete mixes in order to ensure a durable final product. The task of air-void system characterization has fallen on the microscopist, who, according to a standard test method laid forth by the American Society of Testing and Materials, must meticulously count or measure about a thousand air-voids per sample as exposed on a cut and polished cross-section of concrete. The equipment used to perform this task has traditionally included a stereomicroscope, a mechanical stage, and a tally counter. Over the past 30 years, with the availability of computers and digital imaging, automated methods have been introduced to perform the same task, but using the same basic equipment. The method described here replaces the microscope and mechanical stage with an ordinary flatbed desktop scanner, and replaces the microscopist and tally counter with a personal computer; two pieces of equipment much more readily available than a microscope with a mechanical stage, and certainly easier to find than a person willing to sit for extended periods of time counting air-voids. Most laboratories that perform air-void system characterization typically have cabinets full of prepared samples with corresponding results from manual operators. Proponents of automated methods often take advantage of this fact by analyzing the same samples and comparing the results. A similar iterative approach is described here where scanned images collected from a significant number of samples are analyzed, the results compared to those of the manual operator, and the settings optimized to best approximate the results of the manual operator. The results of this calibration procedure are compared to an alternative calibration procedure based on the more rigorous digital image accuracy

  15. The use of thrombin in the radiology department.

    LENUS (Irish Health Repository)

    Ward, E

    2009-03-01

    Thrombin is a naturally occurring coagulation protein that converts soluble fibrinogen into insoluble fibrin and plays a vital role in the coagulation cascade and in turn haemostasis. Thrombin also promotes platelet activation. In the last few years, there has been a rapid increase in the use of thrombin by radiologists in a variety of clinical circumstances. It is best known for its use in the treatment of pseudoaneurysms following angiography. However, there are now a variety of cases in the literature describing the treatment of traumatic, inflammatory and infected aneurysms with thrombin in a variety of locations within the human body. There have even been recent reports describing the use of thrombin in conventional aneurysms as well as ruptured aneurysms. Its use has also been described in the treatment of endoleaks (type II) following aneurysm repair. In nearly all of these cases, treatment with thrombin requires imaging guidance. Recently, thrombin has also been used as a topical treatment post-percutaneous intervention to reduce or stop bleeding. Most radiologists have only a limited knowledge of the pharmacodynamics of thrombin, its wide range of utilisation and its limitations. Apart from a few case reports and case series, there is little in the radiological literature encompassing the wide range of applications that thrombin may have in the radiology department. In this review article, we comprehensively describe the role and pathophysiology of thrombin, describing with examples many of its potential uses. Techniques of usage as well as pitfalls and limitations are also described.

  16. The use of thrombin in the radiology department

    Energy Technology Data Exchange (ETDEWEB)

    Ward, E.; Buckley, O.; Browne, R.F. [Adelaide and Meath Hospitals incorporating the National Children' s Hospital (AMNCH), Department of Radiology, Dublin 24 (Ireland); Collins, A. [Royal Victoria Hospital, Department of Radiology, Belfast (United Kingdom); Torreggiani, W.C. [Adelaide and Meath Hospitals incorporating the National Children' s Hospital (AMNCH), Department of Radiology, Dublin 24 (Ireland)]|[Adelaide and Meath Hospital, Department of Radiology, Dublin 24 (Ireland)

    2009-03-15

    Thrombin is a naturally occurring coagulation protein that converts soluble fibrinogen into insoluble fibrin and plays a vital role in the coagulation cascade and in turn haemostasis. Thrombin also promotes platelet activation. In the last few years, there has been a rapid increase in the use of thrombin by radiologists in a variety of clinical circumstances. It is best known for its use in the treatment of pseudoaneurysms following angiography. However, there are now a variety of cases in the literature describing the treatment of traumatic, inflammatory and infected aneurysms with thrombin in a variety of locations within the human body. There have even been recent reports describing the use of thrombin in conventional aneurysms as well as ruptured aneurysms. Its use has also been described in the treatment of endoleaks (type II) following aneurysm repair. In nearly all of these cases, treatment with thrombin requires imaging guidance. Recently, thrombin has also been used as a topical treatment post-percutaneous intervention to reduce or stop bleeding. Most radiologists have only a limited knowledge of the pharmacodynamics of thrombin, its wide range of utilisation and its limitations. Apart from a few case reports and case series, there is little in the radiological literature encompassing the wide range of applications that thrombin may have in the radiology department. In this review article, we comprehensively describe the role and pathophysiology of thrombin, describing with examples many of its potential uses. Techniques of usage as well as pitfalls and limitations are also described. (orig.)

  17. Thrombin-induced increase in albumin permeability across the endothelium

    International Nuclear Information System (INIS)

    Garcia, J.G.; Siflinger-Birnboim, A.; Bizios, R.; Del Vecchio, P.J.; Fenton, J.W. II; Malik, A.B.

    1986-01-01

    We studied the effect of thrombin on albumin permeability across the endothelial monolayer in vitro. Bovine pulmonary artery endothelial cells were grown on micropore membranes. Morphologic analysis confirmed the presence of a confluent monolayer with interendothelial junctions. Albumin permeability was measured by the clearance of 125I-albumin across the endothelial monolayer. The control 125I-albumin clearance was 0.273 +/- 0.02 microliter/min. The native enzyme, alpha-thrombin (10(-6) to 10(-10) M), added to the luminal side of the endothelium produced concentration-dependent increases in albumin clearance (maximum clearance of 0.586 +/- 0.08 microliter/min at 10(-6) M). Gamma (gamma) thrombin (10(-6) M and 10(-8) M), which lacks the fibrinogen recognition site, also produced a concentration-dependent increase in albumin clearance similar to that observed with alpha-thrombin. Moreover, the two proteolytically inactive forms of the native enzyme, i-Pr2 P-alpha-thrombin and D-Phe-Pro-Arg-CH2-alpha-thrombin, increased the 125I-albumin clearance (0.610 +/- 0.09 microliter/min and 0.609 +/- 0.02 microliter/min for i-Pr2 P-alpha-thrombin and D-Phe-Pro-Arg-CH2-alpha-thrombin at 10(-6) M, respectively). Since the modified forms of thrombin lack the fibrinogen recognition and active serine protease sites, the results indicate that neither site is required for increased albumin permeability. The increase in albumin clearance with alpha-thrombin was not secondary to endothelial cell lysis because lactate dehydrogenase concentration in the medium following thrombin was not significantly different from baseline values. There was also no morphological evidence of cell lysis. Moreover, the increase in 125I-albumin clearance induced by alpha-thrombin was reversible by washing thrombin from the endothelium

  18. Automated Gain Control and Internal Calibration With External Ion Accumulation Capillary liquid chromatography-electrospray ionization-fourier transform ion cyclotron resonance.

    Energy Technology Data Exchange (ETDEWEB)

    Belov, Mikhail E.(VISITORS); Zhang, Rui (BATTELLE (PACIFIC NW LAB)); Strittmatter, Eric F.(BATTELLE (PACIFIC NW LAB)); Prior, David C.(BATTELLE (PACIFIC NW LAB)); Tang, Keqi (BATTELLE (PACIFIC NW LAB)); Smith, Richard D.(BATTELLE (PACIFIC NW LAB))

    2003-08-15

    When combined with capillary LC separations, Electrospray Ionization Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (ESI-FTICR MS) has increasingly been applied for advanced characterization of proteolytic digests. Incorporation of external (to the ICR cell) ion accumulation multipoles with FTICR for ion pre selection and accumulation has enhanced the dynamic range, sensitivity and duty cycle of measurements. However, the highly variable ion production rate from an LC separation can result in overfilling of the external trap, resulting in m/z discrimination and fragmentation of peptide ions. An excessive space charge trapped in the ICR cell causes significant shifts in the detected ion cyclotron frequencies, reducing the achievable mass measurement accuracy (MMA) for protein identification. To eliminate m/z discrimination in the external ion trap, further increase the duty cycle and improve MMA, we developed a capability for data-dependent adjustment of ion accumulation times in the course of an LC separation, referred to as Automated Gain Control (AGC), in combination with low kinetic energy gated ion trapping and internal calibration using a dual-channel electrodynamic ion funnel. The system was initially evaluated in the analysis of a 0.5 mg/mL tryptic digest of bovine serum albumin. The implementation of LC/ESI/AGC/FTICR with internal calibration gave rise to a {approx} 10-fold increase in the number of identified tryptic peptides within mass measurement accuracy of 2 ppm as compared to that detected during the conventional LC/FTICR run with a fixed ion accumulation time and external calibration.

  19. THROMBIN GENERATION AND BLEEDING IN HEMOPHILIA A

    Science.gov (United States)

    Brummel-Ziedins, Kathleen E.; Whelihan, Matthew F.; Gissel, Matthew; Mann, Kenneth G.; Rivard, Georges E.

    2012-01-01

    Introduction Hemophilia A displays phenotypic heterogeneity with respect to clinical severity. Aim To determine if tissue factor (TF)-initiated thrombin generation profiles in whole blood in the presence of corn trypsin inhibitor (CTI) are predictive of bleeding risk in hemophilia A. Methods We studied factor(F) VIII deficient individuals (11 mild, 4 moderate and 12 severe) with a well-characterized five-year bleeding history that included hemarthrosis, soft tissue hematoma and annual FVIII concentrate usage. This clinical information was used to generate a bleeding score. The bleeding scores (range 0–32) were separated into three groups (bleeding score groupings: 0, 0 and ≤9.6, >9.6), with the higher bleeding tendency having a higher score. Whole blood collected by phlebotomy and contact pathway suppressed by 100μg/mL CTI was stimulated to react by the addition of 5pM TF. Reactions were quenched at 20min by inhibitors. Thrombin generation, determined by ELISA for thrombin – antithrombin was evaluated in terms of clot time (CT), maximum level (MaxL) and maximum rate (MaxR) and compared to the bleeding score. Results Data are shown as the mean±SD. MaxL was significantly different (phemophilia A. PMID:19563500

  20. Topical thrombin preparations and their use in cardiac surgery

    Directory of Open Access Journals (Sweden)

    Brianne L Dunn

    2009-10-01

    Full Text Available Brianne L Dunn1, Walter E Uber1, John S Ikonomidis21Department of Pharmacy Services and 2Division of Cardiothoracic Surgery, Medical University of South Carolina, Charleston, South Carolina, USAAbstract: Coagulopathic bleeding may lead to increased morbidity and mortality after cardiac surgery. Topical bovine thrombin has been used to promote hemostasis after surgical procedures for over 60 years and is used frequently as a topical hemostatic agent in cardiac surgery. Recently, use of bovine thrombin has been reported to be associated with increased risk for anaphylaxis, thrombosis, and immune-mediated coagulopathy thought secondary to the production of antifactor V and antithrombin antibodies. In patients who develop bovine thrombin-induced immune-mediated coagulopathy, clinical manifestations may range from asymptomatic alterations in coagulation tests to severe hemorrhage and death. Patients undergoing cardiac surgical procedures may be at increased risk for development of antibodies to bovine thrombin products and associated complications. This adverse immunologic profile has led to the development of alternative preparations including a human and a recombinant thrombin which have been shown to be equally efficacious to bovine thrombin and have reduced antigenicity. However, the potential benefit associated with reduced antigenicity is not truly known secondary to the lack of long-term experience with these products. Given the potentially higher margin of safety and less stringent storage concerns compared to human thrombin, recombinant thrombin may be the most reasonable approach in cardiac surgery.Keywords: bovine thrombin, human thrombin, recombinant thrombin, immune-mediated coagulopathy, topical hemostatic agents, thrombin 

  1. Human Thrombin Injection for the Percutaneous Treatment of Iatrogenic Pseudoaneurysms

    International Nuclear Information System (INIS)

    Elford, Julian; Burrell, Christopher; Freeman, Simon; Roobottom, Carl

    2002-01-01

    Purpose: Thrombin injection is becoming well established for the percutaneous management of iatrogenic pseudoaneurysms. All the published series to date use bovine thrombin,and there have been reports of adverse immunologic effects following its use. Our study aimed to assess the efficacy of human thrombin injection for pseudoaneurysm occlusion. Methods:Fourteen patients with iatrogenic pseudoaneurysms underwent a color Doppler ultrasound examination to assess their suitability for percutaneous human thrombin injection. Human thrombin 1000 IU was then injected into the pseudoaneurysm sac under sterile conditions and with ultrasound guidance. A further color Doppler ultrasound examination was performed 24 hr later to confirm occlusion. Results: All 14 pseudoaneurysms were successfully occluded by human thrombin injection. In two cases a second injection of thrombin was required,but there were no other complications, and all pseudoaneurysms remained occluded at 24 hr. Conclusion: Ultrasound-guided human thrombin injection is simple to perform, effective and safe. We recommend that human thrombin becomes the agent of choice for percutaneous injection into iatrogenic pseudoaneurysms

  2. Simulating groundwater flow and runoff for the Oro Moraine aquifer system. Part II. Automated calibration and mass balance calculations

    Science.gov (United States)

    Beckers, J.; Frind, E. O.

    2001-03-01

    A steady-state groundwater model of the Oro Moraine aquifer system in Central Ontario, Canada, is developed. The model is used to identify the role of baseflow in the water balance of the Minesing Swamp, a 70 km 2 wetland of international significance. Lithologic descriptions are used to develop a hydrostratigraphic conceptual model of the aquifer system. The numerical model uses long-term averages to represent temporal variations of the flow regime and includes a mechanism to redistribute recharge in response to near-surface geologic heterogeneity. The model is calibrated to water level and streamflow measurements through inverse modeling. Observed baseflow and runoff quantities validate the water mass balance of the numerical model and provide information on the fraction of the water surplus that contributes to groundwater flow. The inverse algorithm is used to compare alternative model zonation scenarios, illustrating the power of non-linear regression in calibrating complex aquifer systems. The adjoint method is used to identify sensitive recharge areas for groundwater discharge to the Minesing Swamp. Model results suggest that nearby urban development will have a significant impact on baseflow to the swamp. Although the direct baseflow contribution makes up only a small fraction of the total inflow to the swamp, it provides an important steady influx of water over relatively large portions of the wetland. Urban development will also impact baseflow to the headwaters of local streams. The model provides valuable insight into crucial characteristics of the aquifer system although definite conclusions regarding details of its water budget are difficult to draw given current data limitations. The model therefore also serves to guide future data collection and studies of sub-areas within the basin.

  3. Manufacture of small calibre quadruple lamina vascular bypass grafts using a novel automated extrusion-phase-inversion method and nanocomposite polymer.

    Science.gov (United States)

    Sarkar, Sandip; Burriesci, Gaetano; Wojcik, Adam; Aresti, Nicholas; Hamilton, George; Seifalian, Alexander M

    2009-04-16

    Long-term patency of expanded polytetrafluoroethylene (ePTFE) small calibre cardiovascular bypass prostheses (hyperplasia due to low compliance, stimulating the search for elastic alternatives. Wall porosity allows effective post-implantation graft healing, encouraging endothelialisation and a measured fibrovascular response. We have developed a novel poly (carbonate) urethane-based nanocomposite polymer incorporating polyhedral oligomeric silsesquioxane (POSS) nanocages (UCL-NANO) which shows anti-thrombogenicity and biostability. We report an extrusion-phase-inversion technique for manufacturing uniform-walled porous conduits using UCL-NANO. Image analysis-aided wall measurement showed that two uniform wall-thicknesses could be specified. Different coagulant conditions revealed the importance of low-temperature phase-inversion for graft integrity. Although minor reduction of pore-size variation resulted from the addition of ethanol or N,N-dimethylacetamide, high concentrations of ethanol as coagulant did not provide uniform porosity throughout the wall. Tensile testing showed the grafts to be elastic with strength being directly proportional to weight. The ultimate strengths achieved were above those expected from haemodynamic conditions, with anisotropy due to the manufacturing process. Elemental analysis by energy-dispersive X-ray analysis did not show a regional variation of POSS on the lumen or outer surface. In conclusion, the automated vertical extrusion-phase-inversion device can reproducibly fabricate uniform-walled small calibre conduits from UCL-NANO. These elastic microporous grafts demonstrate favourable mechanical integrity for haemodynamic exposure and are currently undergoing in-vivo evaluation of durability and healing properties.

  4. Platelet glycoprotein VI binds to polymerized fibrin and promotes thrombin generation.

    Science.gov (United States)

    Mammadova-Bach, Elmina; Ollivier, Véronique; Loyau, Stéphane; Schaff, Mathieu; Dumont, Bénédicte; Favier, Rémi; Freyburger, Geneviève; Latger-Cannard, Véronique; Nieswandt, Bernhard; Gachet, Christian; Mangin, Pierre H; Jandrot-Perrus, Martine

    2015-07-30

    Fibrin, the coagulation end product, consolidates the platelet plug at sites of vascular injury and supports the recruitment of circulating platelets. In addition to integrin αIIbβ3, another as-yet-unidentified receptor is thought to mediate platelet interaction with fibrin. Platelet glycoprotein VI (GPVI) interacts with collagen and several other adhesive macromolecules. We evaluated the hypothesis that GPVI could be a functional platelet receptor for fibrin. Calibrated thrombin assays using platelet-rich plasma (PRP) showed that tissue factor-triggered thrombin generation was impaired in GPVI-deficient patients and reduced by the anti-GPVI Fab 9O12. Assays on reconstituted PRP and PRP from fibrinogen-deficient patients revealed a fibrinogen-dependent enhancement of thrombin generation, which relied on functional GPVI. The effect of GPVI was found to depend on fibrin polymerization. A binding assay showed a specific interaction between GPVI-Fc and fibrin, inhibited by the Fab 9O12. This Fab also reduced platelet adhesion to fibrin at low (300 s(-1)) and high (1500 s(-1)) wall shear rates. Platelets adherent to fibrin displayed shape change, exposure of procoagulant phospholipids, and the formation of small clots. When hirudinated blood was perfused at 1500 s(-1) over preformed fibrin-rich clots, the Fab 9O12 decreased the recruitment of platelets by up to 85%. This study identifies GPVI as a platelet receptor for polymerized fibrin with 2 major functions: (1) amplification of thrombin generation and (2) recruitment of circulating platelets to clots. These so-far-unrecognized properties of GPVI confer on it a key role in thrombus growth and stabilization. © 2015 by The American Society of Hematology.

  5. Thrombin-receptor antagonist vorapaxar in acute coronary syndromes

    DEFF Research Database (Denmark)

    Tricoci, Pierluigi; Huang, Zhen; Held, Claes

    2012-01-01

    Vorapaxar is a new oral protease-activated-receptor 1 (PAR-1) antagonist that inhibits thrombin-induced platelet activation.......Vorapaxar is a new oral protease-activated-receptor 1 (PAR-1) antagonist that inhibits thrombin-induced platelet activation....

  6. Hypersensitivity to thrombin of platelets from hypercholesterolemic rats

    International Nuclear Information System (INIS)

    Winocour, P.D.; Rand, M.L.; Kinlough-Rathbone, R.L.; Mustard, J.F.

    1986-01-01

    Hypersensitivity of platelets to thrombin has been associated with hypercholesterolemia. The authors have examined the mechanisms involved in this hypersensitivity. Rats were given diets rich in milk fat and containing added cholesterol and taurocholate to produce hypercholesterolemia (HC) (262 +/- 25 mg%) or added sitosterol as a normocholesterolemic control (NC) (89 +/- 6 mg%). Washed platelets were prelabelled with 14 C-serotonin. In the presence of acetylsalicyclic acid (ASA) (to inhibit thromboxane A 2 (TXA 2 ) formation) and creatine phosphate/creatine phosphokinase (CP/CPK) (to remove released ADP), HC platelets aggregated more (26 +/- 1%) and released more 14 C (9.1 +/- 2.0%) than NC platelets (aggregation: 0%, p 14 C release: 1.5 +/- 0.5%, p 2 formation is involved in the hypersensitivity of HC platelets to thrombin. Total binding of 125 I-thrombin to HC platelets was less than that to NC platelets but HC platelets were smaller and had less protein than NC platelets; the thrombin binding per mg platelet protein was the same for HC and NC platelets, indicating that hypersensitivity to thrombin of HC platelets does not result from increased thrombin binding. Thus, hypersensitivity of HC platelets to thrombin is not due to TXA 2 formation, the action of released ADP or increased thrombin binding

  7. 21 CFR 864.7875 - Thrombin time test.

    Science.gov (United States)

    2010-04-01

    ... 21 Food and Drugs 8 2010-04-01 2010-04-01 false Thrombin time test. 864.7875 Section 864.7875 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES (CONTINUED) MEDICAL DEVICES HEMATOLOGY AND PATHOLOGY DEVICES Hematology Kits and Packages § 864.7875 Thrombin time test. (a...

  8. Statins but not aspirin reduce thrombotic risk assessed by thrombin generation in diabetic patients without cardiovascular events: the RATIONAL trial.

    Directory of Open Access Journals (Sweden)

    Alejandro Macchia

    Full Text Available The systematic use of aspirin and statins in patients with diabetes and no previous cardiovascular events is controversial. We sought to assess the effects of aspirin and statins on the thrombotic risk assessed by thrombin generation (TG among patients with type II diabetes mellitus and no previous cardiovascular events.Prospective, randomized, open, blinded to events evaluation, controlled, 2×2 factorial clinical trial including 30 patients randomly allocated to aspirin 100 mg/d, atorvastatin 40 mg/d, both or none. Outcome measurements included changes in TG levels after treatment (8 to 10 weeks, assessed by a calibrated automated thrombogram. At baseline all groups had similar clinical and biochemical profiles, including TG levels. There was no interaction between aspirin and atorvastatin. Atorvastatin significantly reduced TG measured as peak TG with saline (85.09±55.34 nmol vs 153.26±75.55 nmol for atorvastatin and control groups, respectively; p = 0.018. On the other hand, aspirin had no effect on TG (121.51±81.83 nmol vs 116.85±67.66 nmol, for aspirin and control groups, respectively; p = 0.716. The effects of treatments on measurements of TG using other agonists were consistent.While waiting for data from ongoing large clinical randomized trials to definitively outline the role of aspirin in primary prevention, our study shows that among diabetic patients without previous vascular events, statins but not aspirin reduce thrombotic risk assessed by TG.ClinicalTrials.gov NCT00793754.

  9. APTAMER-BASED SERRS SENSOR FOR THROMBIN DETECTION

    Energy Technology Data Exchange (ETDEWEB)

    Cho, H; Baker, B R; Wachsmann-Hogiu, S; Pagba, C V; Laurence, T A; Lane, S M; Lee, L P; Tok, J B

    2008-07-02

    We describe an aptamer-based Surface Enhanced Resonance Raman Scattering (SERRS) sensor with high sensitivity, specificity, and stability for the detection of a coagulation protein, human a-thrombin. The sensor achieves high sensitivity and a limit of detection of 100 pM by monitoring the SERRS signal change upon the single step of thrombin binding to immobilized thrombin binding aptamer. The selectivity of the sensor is demonstrated by the specific discrimination of thrombin from other protein analytes. The specific recognition and binding of thrombin by the thrombin binding aptamer is essential to the mechanism of the aptamer-based sensor, as shown through measurements using negative control oligonucleotides. In addition, the sensor can detect 1 nM thrombin in the presence of complex biofluids, such as 10% fetal calf serum, demonstrating that the immobilized, 5{prime}-capped, 3{prime}-capped aptamer is sufficiently robust for clinical diagnostic applications. Furthermore, the proposed sensor may be implemented for multiplexed detection using different aptamer-Raman probe complexes.

  10. The interaction of thrombin with platelet protease nexin

    International Nuclear Information System (INIS)

    Knupp, C.L.

    1989-01-01

    Thrombin interacts with a platelet protein which is immunologically related to fibroblast protease nexin and has been termed platelet protease nexin I (PNI). Conflicting hypotheses about the relationship of the thrombin-PNI complex formation to platelet activation have been proposed. The studies presented here demonstrate that the platelet-associated and supernatant complexes with added 125I-thrombin are formed only under conditions which produce platelet activation in normal and chymotrypsin-modified platelets. The platelet-associated complex is formed prior to the appearance of complexes in supernatants. Appearance of the supernatant complex coincides with the appearance of thrombospondin in the reaction supernatants. Excess native thrombin, dansylarginine N-(3-ethyl-1,5-pentanediyl) amide or hirudin can prevent radiolabeled platelet-associated complex formation if added before 125I-thrombin. DAPA or hirudin can prevent or dissociate complex formation if added up to one minute after thrombin but not at later time points. The surface associated complex is accessible to trypsin although a portion remains with the cytoskeletal proteins when thrombin-activated platelets are solubilized with Triton X 100. The surface-associated complex formation parallels many aspects of the specific measurable thrombin binding, yet it does not appear to involve other identified surface glycoprotein thrombin receptors or substrates. Although the time course of appearance of the complexes in supernatants is consistent with other data which suggest that PNI may be released from platelet granules during platelet activation, other explanations for the appearance of PNI on the platelet surface and in supernatants during platelet activation are possible

  11. Label-free aptamer biosensor for selective detection of thrombin

    Energy Technology Data Exchange (ETDEWEB)

    Na, Weidan; Liu, Xiaotong; Wang, Lei; Su, Xingguang, E-mail: suxg@jlu.edu.cn

    2015-10-29

    We fabricated a novel fluorescence biosensor for the selective detection of thrombin by using bovine serum albumin-capped CdS quantum dots (BSA-CdS QDs). Two kinds of designed DNA (DNA1 and DNA2) could bind to CdS QDs through the electrostatic interaction between DNA and Cd{sup 2+} on the surface of CdS QDs. The obtained DNA/BSA-CdS QDs kept stable in the solution with the fluorescence intensity obviously enhanced. Hairpin structure of DNA1contained two domains, one is the aptamer sequence of thrombin and the other is the complementary sequence of DNA2. When thrombin was added, it would bind to DNA1 and induce the hairpin structure of DNA1 changed into G-quadplex structure. Meanwhile, DNA2 would transfer from the surface of CdS QDs to DNA1 via hybridization, which resulted in the removal of DNA1 and DNA2 from the surface of CdS QDs, and led to the fluorescence intensity of CdS QDs reduced. Thus, the determination of thrombin could be achieved by monitoring the change of the fluorescence intensity of CdS QDs. The present method is simple and fast, and exhibits good selectivity for thrombin over other proteins. We have successfully detected thrombin in human serum samples with satisfactory results. - Highlights: • A novel strategy for the detection of thrombin was established based on BSA-CdS QDs. • DNA could serve as the co-ligands to stabilize CdS QDs and enhance the fluorescence intensity. • Thrombin could change the structure of DNA1 and quench the fluorescence of CdS QDs. • Thrombin in real sample was detected with satisfactory results.

  12. Thrombin binding to human brain and spinal cord

    International Nuclear Information System (INIS)

    McKinney, M.; Snider, R.M.; Richelson, E.

    1983-01-01

    Thrombin, a serine protease that regulates hemostasis, has been shown to stimulate the formation of cGMP in murine neuroblastoma cells. The nervous system in vivo thus may be postulated to respond to this blood-borne factor after it breaches the blood-brain barrier, as in trauma. Human alpha-thrombin was radiolabeled with 125I and shown to bind rapidly, reversibly, and with high affinity to human brain and spinal cord. These findings indicate the presence of specific thrombin-binding sites in nervous tissue and may have important clinical implications

  13. A review of three stand-alone topical thrombins for surgical hemostasis.

    Science.gov (United States)

    Cheng, Christine M; Meyer-Massetti, Carla; Kayser, Steven R

    2009-01-01

    Topical thrombins are active hemostatic agents that can be used to minimize blood loss during surgery. Before 2007, the only topical thrombins available were derived from bovine plasma. Antibody formation to bovine thrombin and/or factor V, with subsequent risk of cross-reactivity with human factor V, and hemorrhagic complications associated with human factor-V deficiencies have been described in case reports of surgeries in which bovine thrombins were used. This risk is now included in the boxed warning section of the bovine thrombin prescribing information. In 2007 and 2008, 2 new topical thrombins from nonbovine sources received approval for use from the US Food and Drug Administration. The 3 active topical thrombins that are currently marketed are bovine plasma-derived thrombin, human plasma-derived thrombin, and human recombinant thrombin. The purpose of this review was to evaluate the literature on the efficacy and safety of topical thrombins and discuss the pharmacoeconomic considerations associated with their use. PubMed, EMBASE, and International Pharmaceutical Abstracts were searched for relevant papers published in English through October 10,2008, using the terms thrombin, human recombinant thrombin, bovine thrombin, plasma derived thrombin, and topical thrombin. Manufacturer-provided materials were also reviewed. Abstracts and unpublished data, as well as evaluations of sealants, adhesives, glues, and other hemostats that contain thrombin mixed with fibrinogen and other clotting factors, were excluded. Four randomized, double-blind studies involving the active, stand-alone topical thrombins were found. The bovine thrombin involved in these studies was the predecessor to the currently marketed, highly purified bovine formulation. No studies comparing the human products, studies involving the highly purified bovine preparation, or placebo-controlled studies involving bovine thrombin were found. In a Phase III comparison of human recombinant thrombin and

  14. The Characteristics of Thrombin in Osteoarthritic Pathogenesis and Treatment

    Directory of Open Access Journals (Sweden)

    Pei-Yu Chou

    2014-01-01

    Full Text Available Osteoarthritis (OA is a mechanical abnormality associated with degradation of joints. It is characterized by chronic, progressive degeneration of articular cartilage, abnormalities of bone, and synovial change. The most common symptom of OA is local inflammation resulting from exogenous stress or endogenous abnormal cytokines. Additionally, OA is associated with local and/or systemic activation of coagulation and anticoagulation pathways. Thrombin plays an important role in the stimulation of fibrin deposition and the proinflammatory processes in OA. Thrombin mediates hemostatic and inflammatory responses and guides the immune response to tissue damage. Thrombin activates intracellular signaling pathways by interacting with transmembrane domain G protein coupled receptors (GPCRs, known as protease-activated receptors (PARs. In pathogenic mechanisms, PARs have been implicated in the development of acute and chronic inflammatory responses in OA. Therefore, discovery of thrombin signaling pathways would help us to understand the mechanism of OA pathogenesis and lead us to develop therapeutic drugs in the future.

  15. Thrombin Generating Capacity and Phenotypic Association in ABO Blood Groups.

    Science.gov (United States)

    Kremers, Romy M W; Mohamed, Abdulrahman B O; Pelkmans, Leonie; Hindawi, Salwa; Hemker, H Coenraad; de Laat, H Bas; Huskens, Dana; Al Dieri, Raed

    2015-01-01

    Individuals with blood group O have a higher bleeding risk than non-O blood groups. This could be explained by the lower levels of FVIII and von Willebrand Factor (VWF) levels in O individuals. We investigated the relationship between blood groups, thrombin generation (TG), prothrombin activation and thrombin inactivation. Plasma levels of VWF, FVIII, antithrombin, fibrinogen, prothrombin and α2Macroglobulin (α2M) levels were determined. TG was measured in platelet rich (PRP) and platelet poor plasma (PPP) of 217 healthy donors and prothrombin conversion and thrombin inactivation were calculated. VWF and FVIII levels were lower (75% and 78%) and α2M levels were higher (125%) in the O group. TG is 10% lower in the O group in PPP and PRP. Less prothrombin was converted in the O group (86%) and the thrombin decay capacity was lower as well. In the O group, α2M plays a significantly larger role in the inhibition of thrombin (126%). In conclusion, TG is lower in the O group due to lower prothrombin conversion, and a larger contribution of α2M to thrombin inactivation. The former is unrelated to platelet function because it is similar in PRP and PPP, but can be explained by the lower levels of FVIII.

  16. New synthetic thrombin inhibitors: molecular design and experimental verification.

    Science.gov (United States)

    Sinauridze, Elena I; Romanov, Alexey N; Gribkova, Irina V; Kondakova, Olga A; Surov, Stepan S; Gorbatenko, Aleksander S; Butylin, Andrey A; Monakov, Mikhail Yu; Bogolyubov, Alexey A; Kuznetsov, Yuryi V; Sulimov, Vladimir B; Ataullakhanov, Fazoyl I

    2011-01-01

    The development of new anticoagulants is an important goal for the improvement of thromboses treatments. The design, synthesis and experimental testing of new safe and effective small molecule direct thrombin inhibitors for intravenous administration. Computer-aided molecular design of new thrombin inhibitors was performed using our original docking program SOL, which is based on the genetic algorithm of global energy minimization in the framework of a Merck Molecular Force Field. This program takes into account the effects of solvent. The designed molecules with the best scoring functions (calculated binding energies) were synthesized and their thrombin inhibitory activity evaluated experimentally in vitro using a chromogenic substrate in a buffer system and using a thrombin generation test in isolated plasma and in vivo using the newly developed model of hemodilution-induced hypercoagulation in rats. The acute toxicities of the most promising new thrombin inhibitors were evaluated in mice, and their stabilities in aqueous solutions were measured. New compounds that are both effective direct thrombin inhibitors (the best K(I) was 50) in the thrombin generation assay of approximately 100 nM) were discovered. These compounds contain one of the following new residues as the basic fragment: isothiuronium, 4-aminopyridinium, or 2-aminothiazolinium. LD(50) values for the best new inhibitors ranged from 166.7 to >1111.1 mg/kg. A plasma-substituting solution supplemented with one of the new inhibitors prevented hypercoagulation in the rat model of hemodilution-induced hypercoagulation. Activities of the best new inhibitors in physiological saline (1 µM solutions) were stable after sterilization by autoclaving, and the inhibitors remained stable at long-term storage over more than 1.5 years at room temperature and at 4°C. The high efficacy, stability and low acute toxicity reveal that the inhibitors that were developed may be promising for potential medical applications.

  17. A Novel Photoelectrochemical Biosensor for Tyrosinase and Thrombin Detection

    Directory of Open Access Journals (Sweden)

    Jiexia Chen

    2016-01-01

    Full Text Available A novel photoelectrochemical biosensor for step-by-step assay of tyrosinase and thrombin was fabricated based on the specific interactions between the designed peptide and the target enzymes. A peptide chain with a special sequence which contains a positively charged lysine-labeled terminal, tyrosine at the other end and a cleavage site recognized by thrombin between them was designed. The designed peptide can be fixed on surface of the CdTe quantum dots (QDs-modified indium-tin oxide (ITO electrode through electrostatic attraction to construct the photoelectrochemical biosensor. The tyrosinase target can catalyze the oxidization of tyrosine by oxygen into ortho-benzoquinone residues, which results in a decrease in the sensor photocurrent. Subsequently, the cleavage site could be recognized and cut off by another thrombin target, restoring the sensor photocurrent. The decrease or increase of photocurrent in the sensor enables us to assay tyrosinase and thrombin. Thus, the detection of tyrosinase and thrombin can be achieved in the linear range from 2.6 to 32 μg/mL and from 4.5 to 100 μg/mL with detection limits of 1.5 μg/mL and 1.9 μg/mL, respectively. Most importantly, this strategy shall allow us to detect different classes of enzymes simultaneously by designing various enzyme-specific peptide substrates.

  18. A Novel Photoelectrochemical Biosensor for Tyrosinase and Thrombin Detection

    Science.gov (United States)

    Chen, Jiexia; Liu, Yifan; Zhao, Guang-Chao

    2016-01-01

    A novel photoelectrochemical biosensor for step-by-step assay of tyrosinase and thrombin was fabricated based on the specific interactions between the designed peptide and the target enzymes. A peptide chain with a special sequence which contains a positively charged lysine-labeled terminal, tyrosine at the other end and a cleavage site recognized by thrombin between them was designed. The designed peptide can be fixed on surface of the CdTe quantum dots (QDs)-modified indium-tin oxide (ITO) electrode through electrostatic attraction to construct the photoelectrochemical biosensor. The tyrosinase target can catalyze the oxidization of tyrosine by oxygen into ortho-benzoquinone residues, which results in a decrease in the sensor photocurrent. Subsequently, the cleavage site could be recognized and cut off by another thrombin target, restoring the sensor photocurrent. The decrease or increase of photocurrent in the sensor enables us to assay tyrosinase and thrombin. Thus, the detection of tyrosinase and thrombin can be achieved in the linear range from 2.6 to 32 μg/mL and from 4.5 to 100 μg/mL with detection limits of 1.5 μg/mL and 1.9 μg/mL, respectively. Most importantly, this strategy shall allow us to detect different classes of enzymes simultaneously by designing various enzyme-specific peptide substrates. PMID:26805846

  19. A fully automated calibration method for an optical see-through head-mounted operating microscope with variable zoom and focus.

    Science.gov (United States)

    Figl, Michael; Ede, Christopher; Hummel, Johann; Wanschitz, Felix; Ewers, Rolf; Bergmann, Helmar; Birkfellner, Wolfgang

    2005-11-01

    Ever since the development of the first applications in image-guided therapy (IGT), the use of head-mounted displays (HMDs) was considered an important extension of existing IGT technologies. Several approaches to utilizing HMDs and modified medical devices for augmented reality (AR) visualization were implemented. These approaches include video-see through systems, semitransparent mirrors, modified endoscopes, and modified operating microscopes. Common to all these devices is the fact that a precise calibration between the display and three-dimensional coordinates in the patient's frame of reference is compulsory. In optical see-through devices based on complex optical systems such as operating microscopes or operating binoculars-as in the case of the system presented in this paper-this procedure can become increasingly difficult since precise camera calibration for every focus and zoom position is required. We present a method for fully automatic calibration of the operating binocular Varioscope M5 AR for the full range of zoom and focus settings available. Our method uses a special calibration pattern, a linear guide driven by a stepping motor, and special calibration software. The overlay error in the calibration plane was found to be 0.14-0.91 mm, which is less than 1% of the field of view. Using the motorized calibration rig as presented in the paper, we were also able to assess the dynamic latency when viewing augmentation graphics on a mobile target; spatial displacement due to latency was found to be in the range of 1.1-2.8 mm maximum, the disparity between the true object and its computed overlay represented latency of 0.1 s. We conclude that the automatic calibration method presented in this paper is sufficient in terms of accuracy and time requirements for standard uses of optical see-through systems in a clinical environment.

  20. Multiple active forms of thrombin. IV. Relative activities of meizothrombins

    Energy Technology Data Exchange (ETDEWEB)

    Doyle, M.F.; Mann, K.G. (Univ. of Vermont College of Medicine, Burlington (USA))

    1990-06-25

    The prothrombin activation intermediates meizothrombin and meizothrombin(desF1) (meizothrombin that has been autoproteolyzed to remove fragment 1) have been obtained in a relatively pure, active form with minimal autolysis, making them suitable for enzymatic characterization. When compared at equimolar concentrations, alpha-thrombin, fragment 1.2+ alpha-thrombin, meizothrombin(desF1), and meizothrombin have approximately 100, 100, 10, and 1% activity, respectively, toward the macromolecular substrates factor V, fibrinogen, and platelets. The difference in activity of these four enzymes cannot be attributed to alterations in the catalytic triad, as all four enzymes have nearly identical catalytic efficiency toward the chromogenic substrate S2238. Further, the ability of meizothrombin and meizothrombin(desF1) to activate protein C was 75% of the activity exhibited by alpha-thrombin or fragment 1.2+ alpha-thrombin. All four enzymes bind to thrombomodulin, as judged by the enhanced rate of protein C activation upon preincubation of the enzymes with thrombomodulin. The extent of rate enhancement varied, with meizothrombin/thrombomodulin exhibiting only 50% of the alpha-thrombin/thrombomodulin rate. This difference in rate is not due to a decreased affinity of the meizothrombin for thrombomodulin since the apparent dissociation constants for the alpha-thrombin-thrombomodulin complex and the meizothrombin-thrombomodulin complex are virtually identical. The difference in the observed rate is due in part to the higher Km for protein C exhibited by the meizothrombin-thrombomodulin complex. Incubation of the thrombomodulin-enzyme complex with phospholipid vesicles caused an increase in the protein C activation rates. The kinetic constants for protein C activation in the presence of phospholipid are virtually identical for these enzyme-thrombomodulin complexes.

  1. Activation of human factor V by factor Xa and thrombin

    International Nuclear Information System (INIS)

    Monkovic, D.D.; Tracy, P.B.

    1990-01-01

    The activation of human factor V by factor Xa and thrombin was studied by functional assessment of cofactor activity and sodium dodecyl sulfate-polycarylamide gel electrophoresis followed by either autoradiography of 125 I-labeled factor V activation products or Western blot analyses of unlabeled factor V activation products. Cofactor activity was measured by the ability of the factor V/Va peptides to support the activation of prothrombin. The factor Xa catalyzed cleavage of factor V was observed to be time, phospholipid, and calcium ion dependent, yielding a cofactor with activity equal to that of thrombin-activated factor V (factor Va). The cleavage pattern differed markedly from the one observed in the bovine system. The factor Xa activated factor V subunits expressing cofactor activity were isolated and found to consist of peptides of M r 220,000 and 105,000. Although thrombin cleaved the M r 220,000 peptide to yield peptides previously shown to be products of thrombin activation, cofactor activity did not increase. N-Terminal sequence analysis confirmed that both factor Xa and thrombin cleave factor V at the same bond to generate the M r 220,000 peptide. The factor Xa dependent functional assessment of 125 I-labeled factor V coupled with densitometric analyses of the cleavage products indicated that the cofactor activity of factor Xa activated factor V closely paralleled the appearance of the M r 220,000 peptide. The data indicate that factor Xa is as efficient an enzyme toward factor V as thrombin

  2. Multiple active forms of thrombin. IV. Relative activities of meizothrombins

    International Nuclear Information System (INIS)

    Doyle, M.F.; Mann, K.G.

    1990-01-01

    The prothrombin activation intermediates meizothrombin and meizothrombin(desF1) (meizothrombin that has been autoproteolyzed to remove fragment 1) have been obtained in a relatively pure, active form with minimal autolysis, making them suitable for enzymatic characterization. When compared at equimolar concentrations, alpha-thrombin, fragment 1.2+ alpha-thrombin, meizothrombin(desF1), and meizothrombin have approximately 100, 100, 10, and 1% activity, respectively, toward the macromolecular substrates factor V, fibrinogen, and platelets. The difference in activity of these four enzymes cannot be attributed to alterations in the catalytic triad, as all four enzymes have nearly identical catalytic efficiency toward the chromogenic substrate S2238. Further, the ability of meizothrombin and meizothrombin(desF1) to activate protein C was 75% of the activity exhibited by alpha-thrombin or fragment 1.2+ alpha-thrombin. All four enzymes bind to thrombomodulin, as judged by the enhanced rate of protein C activation upon preincubation of the enzymes with thrombomodulin. The extent of rate enhancement varied, with meizothrombin/thrombomodulin exhibiting only 50% of the alpha-thrombin/thrombomodulin rate. This difference in rate is not due to a decreased affinity of the meizothrombin for thrombomodulin since the apparent dissociation constants for the alpha-thrombin-thrombomodulin complex and the meizothrombin-thrombomodulin complex are virtually identical. The difference in the observed rate is due in part to the higher Km for protein C exhibited by the meizothrombin-thrombomodulin complex. Incubation of the thrombomodulin-enzyme complex with phospholipid vesicles caused an increase in the protein C activation rates. The kinetic constants for protein C activation in the presence of phospholipid are virtually identical for these enzyme-thrombomodulin complexes

  3. New synthetic thrombin inhibitors: molecular design and experimental verification.

    Directory of Open Access Journals (Sweden)

    Elena I Sinauridze

    Full Text Available BACKGROUND: The development of new anticoagulants is an important goal for the improvement of thromboses treatments. OBJECTIVES: The design, synthesis and experimental testing of new safe and effective small molecule direct thrombin inhibitors for intravenous administration. METHODS: Computer-aided molecular design of new thrombin inhibitors was performed using our original docking program SOL, which is based on the genetic algorithm of global energy minimization in the framework of a Merck Molecular Force Field. This program takes into account the effects of solvent. The designed molecules with the best scoring functions (calculated binding energies were synthesized and their thrombin inhibitory activity evaluated experimentally in vitro using a chromogenic substrate in a buffer system and using a thrombin generation test in isolated plasma and in vivo using the newly developed model of hemodilution-induced hypercoagulation in rats. The acute toxicities of the most promising new thrombin inhibitors were evaluated in mice, and their stabilities in aqueous solutions were measured. RESULTS: New compounds that are both effective direct thrombin inhibitors (the best K(I was 1111.1 mg/kg. A plasma-substituting solution supplemented with one of the new inhibitors prevented hypercoagulation in the rat model of hemodilution-induced hypercoagulation. Activities of the best new inhibitors in physiological saline (1 µM solutions were stable after sterilization by autoclaving, and the inhibitors remained stable at long-term storage over more than 1.5 years at room temperature and at 4°C. CONCLUSIONS: The high efficacy, stability and low acute toxicity reveal that the inhibitors that were developed may be promising for potential medical applications.

  4. Increased anticoagulant activity of thrombin-binding DNA aptamers by nanoscale organization on DNA nanostructures

    DEFF Research Database (Denmark)

    Rangnekar, Abhijit; Zhang, Alex M.; Shiyuan Li, Susan

    2012-01-01

    Control over thrombin activity is much desired to regulate blood clotting in surgical and therapeutic situations. Thrombin-binding RNA and DNA aptamers have been used to inhibit thrombin activity and thus the coagulation cascade. Soluble DNA aptamers, as well as two different aptamers tethered by...

  5. Thrombin regulates components of the fibrinolytic system in human mesangial cells

    International Nuclear Information System (INIS)

    Villamediana, L.M.; Rondeau, E.; He, C.J.; Medcalf, R.L.; Peraldi, M.N.; Lacave, R.; Delarue, F.; Sraer, J.D.

    1990-01-01

    Besides its procoagulant activity, thrombin has been shown to stimulate cell proliferation and to regulate the fibrinolytic pathway. We report here the effect of purified human alpha thrombin on the synthesis of tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor 1 (PAI-1) by cultured human mesangial cells. Thrombin (0 to 2.5 U/ml) increased in a time- and dose-dependent manner the production of t-PA and PAI-1 (2- to 3-fold increase of secreted t-PA and PAI-1 release during a 24 hour incubation). This effect was associated with a twofold increase in DNA synthesis measured by 3H-thymidine incorporation. Zymographic analysis and reverse fibrin autography showed that thrombin also increased the level of the 110 Kd t-PA-PAI-1 complex, whereas PAI-1 was present as a free 50 Kd form in the culture medium conditioned by unstimulated and thrombin-stimulated cells. Free t-PA was never observed. Both membrane binding and catalytic activity of thrombin were required since the effects of 1 U/ml thrombin were inhibited by addition 2 U/ml hirudin, which inhibits the membrane binding and catalytic activity of thrombin, and since DFP-inactivated thrombin, which has the ability to bind but which has no enzymatic activity, did not induce t-PA or PAI-1. Gamma thrombin, which does not bind to thrombin receptor, did not increase t-PA and PAI-1 releases. The effects of thrombin were probably mediated by protein kinase C activation since H7, an inhibitor of protein kinases, inhibited significantly thrombin effects on t-PA and PAI-1 production, and since addition of an activator of protein kinase A, 8-bromocyclic AMP (100 microM), induced a significant inhibition of the thrombin effect. The effects of thrombin were also suppressed by 1.25 micrograms/ml alpha amanitin, suggesting a requirement of de novo RNA synthesis

  6. Thrombin induces rapid PAR1-mediated non-classical FGF1 release

    International Nuclear Information System (INIS)

    Duarte, Maria; Kolev, Vihren; Soldi, Raffaella; Kirov, Alexander; Graziani, Irene; Oliveira, Silvia Marta; Kacer, Doreen; Friesel, Robert; Maciag, Thomas; Prudovsky, Igor

    2006-01-01

    Thrombin induces cell proliferation and migration during vascular injury. We report that thrombin rapidly stimulated expression and release of the pro-angiogenic polypeptide fibroblast growth factor 1 (FGF1). Thrombin failed to induce FGF1 release from protease-activated receptor 1 (PAR1) null fibroblasts, indicating that this effect was dependent on PAR1. Similarly to thrombin, FGF1 expression and release were induced by TRAP, a specific oligopeptide agonist of PAR1. These results identify a novel aspect of the crosstalk between FGF and thrombin signaling pathways which both play important roles in tissue repair and angiogenesis

  7. Rigidification of the autolysis loop enhances Na(+) binding to thrombin.

    Science.gov (United States)

    Pozzi, Nicola; Chen, Raymond; Chen, Zhiwei; Bah, Alaji; Di Cera, Enrico

    2011-11-01

    Binding of Na(+) to thrombin ensures high activity toward physiological substrates and optimizes the procoagulant and prothrombotic roles of the enzyme in vivo. Under physiological conditions of pH and temperature, the binding affinity of Na(+) is weak due to large heat capacity and enthalpy changes associated with binding, and the K(d)=80 mM ensures only 64% saturation of the site at the concentration of Na(+) in the blood (140 mM). Residues controlling Na(+) binding and activation have been identified. Yet, attempts to improve the interaction of Na(+) with thrombin and possibly increase catalytic activity under physiological conditions have so far been unsuccessful. Here we report how replacement of the flexible autolysis loop of human thrombin with the homologous rigid domain of the murine enzyme results in a drastic (up to 10-fold) increase in Na(+) affinity and a significant improvement in the catalytic activity of the enzyme. Rigidification of the autolysis loop abolishes the heat capacity change associated with Na(+) binding observed in the wild-type and also increases the stability of thrombin. These findings have general relevance to protein engineering studies of clotting proteases and trypsin-like enzymes. Copyright © 2011 Elsevier B.V. All rights reserved.

  8. Rigidification of the autolysis loop enhances Na+ binding to thrombin

    Science.gov (United States)

    Pozzi, Nicola; Chen, Raymond; Chen, Zhiwei; Bah, Alaji; Di Cera, Enrico

    2011-01-01

    Binding of Na+ to thrombin ensures high activity toward physiological substrates and optimizes the procoagulant and prothrombotic roles of the enzyme in vivo. Under physiological conditions of pH and temperature, the binding affinity of Na+ is weak due to large heat capacity and enthalpy changes associated with binding, and the Kd=80 mM ensures only 64% saturation of the site at the concentration of Na+ in the blood (140 mM). Residues controlling Na+ binding and activation have been identified. Yet, attempts to improve the interaction of Na+ with thrombin and possibly increase catalytic activity under physiological conditions have so far been unsuccessful. Here we report how replacement of the flexible autolysis loop of human thrombin with the homologous rigid domain of the murine enzyme results in a drastic (up to 10-fold) increase in Na+ affinity and a significant improvement in the catalytic activity of the enzyme. Rigidification of the autolysis loop abolishes the heat capacity change associated with Na+ binding observed in the wild-type and also increases the stability of thrombin. These findings have general relevance to protein engineering studies of clotting proteases and trypsin-like enzymes. PMID:21536369

  9. Tyrosine sulfation modulates activity of tick-derived thrombin inhibitors

    Science.gov (United States)

    Thompson, Robert E.; Liu, Xuyu; Ripoll-Rozada, Jorge; Alonso-García, Noelia; Parker, Benjamin L.; Pereira, Pedro José Barbosa; Payne, Richard J.

    2017-09-01

    Madanin-1 and chimadanin are two small cysteine-free thrombin inhibitors that facilitate blood feeding in the tick Haemaphysalis longicornis. Here, we report a post-translational modification—tyrosine sulfation—of these two proteins that is critical for potent anti-thrombotic and anticoagulant activity. Inhibitors produced in baculovirus-infected insect cells displayed heterogeneous sulfation of two tyrosine residues within each of the proteins. One-pot ligation-desulfurization chemistry enabled access to homogeneous samples of all possible sulfated variants of the proteins. Tyrosine sulfation of madanin-1 and chimadanin proved crucial for thrombin inhibitory activity, with the doubly sulfated variants three orders of magnitude more potent than the unmodified inhibitors. The three-dimensional structure of madanin-1 in complex with thrombin revealed a unique mode of inhibition, with the sulfated tyrosine residues binding to the basic exosite II of the protease. The importance of tyrosine sulfation within this family of thrombin inhibitors, together with their unique binding mode, paves the way for the development of anti-thrombotic drug leads based on these privileged scaffolds.

  10. Increased thrombin generation in women with polycystic ovary syndrome

    DEFF Research Database (Denmark)

    Glintborg, Dorte; Sidelmann, Johannes Jakobsen; Lambaa Altinok, Magda

    2015-01-01

    Objective. Polycystic ovary syndrome (PCOS) is associated with risk factors for cardiovascular disease (CVD) which may be modified by the use of metformin and oral contraceptives (OC). Thrombin generation (TG) measures are risk markers of CVD and address the composite of multiple factors...

  11. Stabilization of the E* Form Turns Thrombin into an Anticoagulant

    Energy Technology Data Exchange (ETDEWEB)

    Bah, Alaji; Carrell, Christopher J.; Chen, Zhiwei; Gandhi, Prafull S.; Di Cera, Enrico; (WU-MED)

    2009-07-31

    Previous studies have shown that deletion of nine residues in the autolysis loop of thrombin produces a mutant with an anticoagulant propensity of potential clinical relevance, but the molecular origin of the effect has remained unresolved. The x-ray crystal structure of this mutant solved in the free form at 1.55 {angstrom} resolution reveals an inactive conformation that is practically identical (root mean square deviation of 0.154 {angstrom}) to the recently identified E* form. The side chain of Trp215 collapses into the active site by shifting >10 {angstrom} from its position in the active E form, and the oxyanion hole is disrupted by a flip of the Glu192-Gly193 peptide bond. This finding confirms the existence of the inactive form E* in essentially the same incarnation as first identified in the structure of the thrombin mutant D102N. In addition, it demonstrates that the anticoagulant profile often caused by a mutation of the thrombin scaffold finds its likely molecular origin in the stabilization of the inactive E* form that is selectively shifted to the active E form upon thrombomodulin and protein C binding.

  12. Aptamer-crosslinked microbubbles: smart contrast agents for thrombin-activated ultrasound imaging.

    Science.gov (United States)

    Nakatsuka, Matthew A; Mattrey, Robert F; Esener, Sadik C; Cha, Jennifer N; Goodwin, Andrew P

    2012-11-27

    Thrombosis, or malignant blood clotting, is associated with numerous cardiovascular diseases and cancers. A microbubble contrast agent is presented that produces ultrasound harmonic signal only when exposed to elevated thrombin levels. Initially silent microbubbles are activated in the presence of both thrombin-spiked and freshly clotting blood in three minutes with detection limits of 20 nM thrombin and 2 aM microbubbles. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  13. Calibration uncertainty

    DEFF Research Database (Denmark)

    Heydorn, Kaj; Anglov, Thomas

    2002-01-01

    Methods recommended by the International Standardization Organisation and Eurachem are not satisfactory for the correct estimation of calibration uncertainty. A novel approach is introduced and tested on actual calibration data for the determination of Pb by ICP-AES. The improved calibration...

  14. Investigation of Interactions between Thrombin and Ten Phenolic Compounds by Affinity Capillary Electrophoresis and Molecular Docking

    Directory of Open Access Journals (Sweden)

    Qiao-Qiao Li

    2018-01-01

    Full Text Available Thrombin plays a vital role in blood coagulation, which is a key process involved in thrombosis by promoting platelet aggregation and converting fibrinogen to form the fibrin clot. In the receptor concept, drugs produce their therapeutic effects via interactions with the targets. Therefore, investigation of interaction between thrombin and small molecules is important to find out the potential thrombin inhibitor. In this study, affinity capillary electrophoresis (ACE and in silico molecular docking methods were developed to study the interaction between thrombin and ten phenolic compounds (p-hydroxybenzoic acid, protocatechuic acid, vanillic acid, gallic acid, catechin, epicatechin, dihydroquercetin, naringenin, apigenin, and baicalein. The ACE results showed that gallic acids and six flavonoid compounds had relative strong interactions with thrombin. In addition, the docking results indicated that all of optimal conformations of the six flavonoid compounds were positioned into the thrombin activity centre and had interaction with the HIS57 or SER195 which was the key residue to bind thrombin inhibitors such as argatroban. Herein, these six flavonoid compounds might have the potential of thrombin inhibition activity. In addition, the developed method in this study can be further applied to study the interactions of other molecules with thrombin.

  15. Percutaneous Ultrasound-Guided Thrombin Injection in Iatrogenic Arterial Pseudoaneurysms: Effectiveness and Complications

    International Nuclear Information System (INIS)

    Koh, Young Hwan; Kim, Hak Soo; Kim, Hyung Sik; Min, Seung Kee

    2005-01-01

    To evaluate and describe the efficacy and side effects of a percutaneous thrombin injection under ultrasonography guidance for the treatment of iatrogenic pseudo aneurysms Eighteen consecutive iatrogenic pseudo aneurysm cases were treated with a thrombin injection. The thrombin was injected into the pseudo aneurysm cavity using a 22-gauge needle under ultrasonographic guidance. The causes of the pseudo aneurysms are as follows: post coronary angiography (9 cases), percutaneous coronary balloon angioplasty (5 cases), cerebral angiography (1 case), transhepatic chemo embolization (1 case), percutaneous trans femoral arterial stent insertion (1 case) and bone marrow aspiration for a marrow transplant (1 case). Only one case required a secondary thrombin injection due to recurrent flow in the pseudo aneurysm lumen, which was detected at the follow up Doppler ultrasound. Other seventeen cases were successfully treated on the first trial. There were no technical failures or complication related to the procedure. The average amount of thrombin injected was 733 IU. Nine out of 18 treated patients (50%) showed mild reactions to the thrombin including mild fever (4 cases), chilling sensation (3 cases), a chilling sensation with mild dyspnea (1 case), mild chest discomfort (1 case) after the thrombin injection. All these side effects were transient and improved several hours later. All the iatrogenic pseudo aneurysms were treated successfully with an ultrasound-guided percutaneous thrombin injection. There was a high rate of hypersensitivity to the bovine thrombin, which precaution should be taken to prevent more serious side effects

  16. Percutaneous Ultrasound-Guided Thrombin Injection in Iatrogenic Arterial Pseudoaneurysms: Effectiveness and Complications

    Energy Technology Data Exchange (ETDEWEB)

    Koh, Young Hwan [Boramae Hospital, Seoul (Korea, Republic of); Kim, Hak Soo; Kim, Hyung Sik; Min, Seung Kee [Gachon Medical School, Incheon (Korea, Republic of)

    2005-09-15

    To evaluate and describe the efficacy and side effects of a percutaneous thrombin injection under ultrasonography guidance for the treatment of iatrogenic pseudo aneurysms Eighteen consecutive iatrogenic pseudo aneurysm cases were treated with a thrombin injection. The thrombin was injected into the pseudo aneurysm cavity using a 22-gauge needle under ultrasonographic guidance. The causes of the pseudo aneurysms are as follows: post coronary angiography (9 cases), percutaneous coronary balloon angioplasty (5 cases), cerebral angiography (1 case), transhepatic chemo embolization (1 case), percutaneous trans femoral arterial stent insertion (1 case) and bone marrow aspiration for a marrow transplant (1 case). Only one case required a secondary thrombin injection due to recurrent flow in the pseudo aneurysm lumen, which was detected at the follow up Doppler ultrasound. Other seventeen cases were successfully treated on the first trial. There were no technical failures or complication related to the procedure. The average amount of thrombin injected was 733 IU. Nine out of 18 treated patients (50%) showed mild reactions to the thrombin including mild fever (4 cases), chilling sensation (3 cases), a chilling sensation with mild dyspnea (1 case), mild chest discomfort (1 case) after the thrombin injection. All these side effects were transient and improved several hours later. All the iatrogenic pseudo aneurysms were treated successfully with an ultrasound-guided percutaneous thrombin injection. There was a high rate of hypersensitivity to the bovine thrombin, which precaution should be taken to prevent more serious side effects

  17. Desenvolvimento e calibração de um tensiômetro eletrônico de leitura automática Development and calibration of an electronic tensiometer for automatic reading

    Directory of Open Access Journals (Sweden)

    Adunias S. Teixeira

    2005-08-01

    Full Text Available O presente trabalho apresenta o desenvolvimento e calibração de tensiômetro de leitura automática, sendo o tensiômetro de mercúrio utilizado como padrão de comparação. Os ensaios foram conduzidos no Laboratório de Mecânica e Eletrônica do Departamento de Engenharia Agrícola da UFC. Esse equipamento difere do tensiômetro tradicional por substituir o manômetro de mercúrio por sensor de pressão. Tal dispositivo gera uma saída com valor de até no máximo 4,5 volts. O tensiômetro eletrônico é conectado a uma placa de aquisição de dados (DAQ contendo um microprocessador, conversor analógico/digital (ADC e saída serial, sendo tal placa ligada a um microcomputador. A bancada de ensaio constituiu-se de três caixas plásticas preenchidas com solo de textura franco- arenosa. Os dados foram coletados durante um mês e as equações geradas foram do tipo Count = offset + apsim, em que psim representa o potencial matricial fornecido pelo tensiômetro de mercúrio (kPa e count a saída digial do ADC, com valores adimensionais de 0 a 4.095. Foram obtidos os valores máximo e mínimo de offset de 348,572 e 261,026, e de coeficiente angular de 36,675 kPa-1 e 34,421 kPa-1. Os resultados da regressão linear simples indicaram a existência de regressão a menos de 0,1% de significância e valores de coeficientes de correlação nunca inferiores a 0,9994.This paper presents the development and calibration of a tensiometer for automatic reading where mercury has been replaced by an electronic pressure sensor called the electronic tensiometer. The conventional mercury tensiometer was used as a standard for calibration. Calibration trials were conducted at the Mechanics and Electronics Laboratory of the Department of Agricultural Engineering at Ceará Federal University, Brazil. The electronic tensiometer outputs a maximum 4.5 volts and is connected to a data acquisition system (DAQ composed of a microprocessor, a 12-bit analog to digital

  18. Calibration factor or calibration coefficient?

    International Nuclear Information System (INIS)

    Meghzifene, A.; Shortt, K.R.

    2002-01-01

    Full text: The IAEA/WHO network of SSDLs was set up in order to establish links between SSDL members and the international measurement system. At the end of 2001, there were 73 network members in 63 Member States. The SSDL network members provide calibration services to end-users at the national or regional level. The results of the calibrations are summarized in a document called calibration report or calibration certificate. The IAEA has been using the term calibration certificate and will continue using the same terminology. The most important information in a calibration certificate is a list of calibration factors and their related uncertainties that apply to the calibrated instrument for the well-defined irradiation and ambient conditions. The IAEA has recently decided to change the term calibration factor to calibration coefficient, to be fully in line with ISO [ISO 31-0], which recommends the use of the term coefficient when it links two quantities A and B (equation 1) that have different dimensions. The term factor should only be used for k when it is used to link the terms A and B that have the same dimensions A=k.B. However, in a typical calibration, an ion chamber is calibrated in terms of a physical quantity such as air kerma, dose to water, ambient dose equivalent, etc. If the chamber is calibrated together with its electrometer, then the calibration refers to the physical quantity to be measured per electrometer unit reading. In this case, the terms referred have different dimensions. The adoption by the Agency of the term coefficient to express the results of calibrations is consistent with the 'International vocabulary of basic and general terms in metrology' prepared jointly by the BIPM, IEC, ISO, OIML and other organizations. The BIPM has changed from factor to coefficient. The authors believe that this is more than just a matter of semantics and recommend that the SSDL network members adopt this change in terminology. (author)

  19. Kinetic modeling sheds light on the mode of action of recombinant factor VIIa on thrombin generation.

    Science.gov (United States)

    Mitrophanov, Alexander Y; Reifman, Jaques

    2011-10-01

    The therapeutic potential of a hemostatic agent can be assessed by investigating its effects on the quantitative parameters of thrombin generation. For recombinant activated factor VII (rFVIIa)--a promising hemostasis-inducing biologic--experimental studies addressing its effects on thrombin generation yielded disparate results. To elucidate the inherent ability of rFVIIa to modulate thrombin production, it is necessary to identify rFVIIa-induced effects that are compatible with the available biochemical knowledge about thrombin generation mechanisms. The existing body of knowledge about coagulation biochemistry can be rigorously represented by a computational model that incorporates the known reactions and parameter values constituting the biochemical network. We used a thoroughly validated numerical model to generate activated factor VII (FVIIa) titration curves in the cases of normal blood composition, hemophilia A and B blood, blood lacking factor VII, blood lacking tissue factor pathway inhibitor, and diluted blood. We utilized the generated curves to perform systematic fold-change analyses for five quantitative parameters characterizing thrombin accumulation. The largest fold changes induced by increasing FVIIa concentration were observed for clotting time, thrombin peak time, and maximum slope of the thrombin curve. By contrast, thrombin peak height was much less affected by FVIIa titrations, and the area under the thrombin curve stayed practically unchanged. Comparisons with experimental data demonstrated that the computationally derived patterns can be observed in vitro. rFVIIa modulates thrombin generation primarily by accelerating the process, without significantly affecting the total amount of generated thrombin. Copyright © 2011 Elsevier Ltd. All rights reserved.

  20. Pulmonary epithelial clearance of 99mTc-DTPA after thrombin-induced pulmonary microembolism

    International Nuclear Information System (INIS)

    Cooper, J.A.; Feustel, P.J.; Line, B.R.; Malik, A.B.

    1986-01-01

    We investigated the effect of thrombin-induced pulmonary microembolism on the pulmonary clearance rate of aerosolized 99mTc diethylenetriamine pentaacetic acid (99mTc-DTPA) in awake, chronically prepared sheep. Chest activity was recorded after administration of a 0.44 micron aerosol of 99mTc-DTPA. Decay-corrected data were fit to an exponential and expressed as percent decrease per min (%/min). Sheep were given alpha-thrombin intravenously (80 U/kg for 10 min) 60 min after the aerosol administration. The clearance rate prior to alpha-thrombin was 0.35 +/- 0.05 %/min (mean +/- SEM). During alpha-thrombin administration, the clearance rate increased to 5.84 +/- 0.70 %/min (p less than 0.001 from baseline), but returned to 0.41 +/- 0.06 %/min within 30 min after the end of the thrombin infusion. The increased clearance rate during alpha-thrombin administration was not due to increased lung volume since alpha-thrombin did not change functional residual capacity. Moreover, the clearance rate was unchanged during gamma-thrombin administration, which does not induce coagulation, or during alpha-thrombin challenge in defibrinogenated animals. alpha-thrombin administration in neutrophil-depleted sheep caused a transient increase in DTPA clearance similar to that in control sheep, suggesting that the increase occurred independently of neutrophils. The results indicate that alpha-thrombin causes a large, transient increase in 99mTc-DTPA clearance, which may be the result of increased epithelial permeability. This response is dependent on the activation of intravascular coagulation

  1. Three different signal amplification strategies for the impedimetric sandwich detection of thrombin

    Energy Technology Data Exchange (ETDEWEB)

    Ocaña, Cristina; Valle, Manel del, E-mail: manel.delvalle@uab.cat

    2016-03-17

    In this work, we report a comparative study on three highly specific amplification strategies for the ultrasensitive detection of thrombin with the use of aptamer sandwich protocol. The protocol consisted on the use of a first thrombin aptamer immobilized on the electrode surface, the recognition of thrombin protein, and the reaction with a second biotinylated thrombin aptamer forming the sandwich. Through the exposed biotin end, three variants have been tested to amplify the electrochemical impedance signal. The strategies included (a) silver enhancement treatment, (b) gold enhancement treatment and (c) insoluble product produced by the combination of the enzyme horseradish peroxidase (HRP) and 3-amino-9-ethylcarbazole (AEC). The properties of the sensing surface were probed by electrochemical impedance measurements in the presence of the ferrocyanide/ferricyanide redox marker. Insoluble product strategy and silver enhancement treatment resulted in the lowest detection limit (0.3 pM), while gold enhancement method resulted in the highest reproducibility, 8.8% RSD at the pM thrombin concentration levels. Results of silver and gold enhancement treatment also permitted direct inspection by scanning electron microscopy (SEM). - Highlights: • Aptasensor to detect thrombin reaching the femtomolar level. • Biosensing protocol employs two thrombin aptamers in a sandwich capture scheme. • Use of second biotinylated aptamer allows many amplification and detection variants. • Precipitation reaction provides the highest signal amplification of ca. 3 times. • Double recognition event improves remarkably selectivity for thrombin detection.

  2. Effects of Aerobic Capacity on Thrombin-Induced Hydrocephalus and White Matter Injury.

    Science.gov (United States)

    Ni, Wei; Gao, Feng; Zheng, Mingzhe; Koch, Lauren G; Britton, Steven L; Keep, Richard F; Xi, Guohua; Hua, Ya

    2016-01-01

    We have previously shown that intracerebral hemorrhage-induced brain injury is less in rats bred for high aerobic capacity (high capacity runners; HCR) compared with those bred for low aerobic capacity (low capacity runners; LCRs). Thrombin, an essential component in the coagulation cascade, is produced after cerebral hemorrhage. Intraventricular injection of thrombin causes significant hydrocephalus and white matter damage. In the present study, we examined the effect of exercise capacity on thrombin-induced hydrocephalus and white matter damage. Mid-aged (13-month-old) female LCRs (n = 13) and HCRs (n = 12) rats were used in this study. Rats received an intraventricular injection of thrombin (3 U, 50 μl). All rats underwent magnetic resonance imaging (MRI) at 24 h and were then euthanized for brain histology and Western blot. The mortalities were 20 % in LCRs and 33 % in HCRs after thrombin injection (p > 0.05). No rats died after saline injection. Intraventricular thrombin injection resulted in hydrocephalus and periventricular white matter damage as determined on MRI. In LCR rats, thrombin induced significant ventricle enlargement (23.0 ± 2.3 vs12.8 ± 1.9 mm(3) in LCR saline group; p hydrocephalus in rats with low aerobic capacity. A differential effect of thrombin may contribute to differences in the effects of cerebral hemorrhage with aerobic capacity.

  3. Dabigatran reduces thrombin-induced platelet aggregation and activation in a dose-dependent manner

    DEFF Research Database (Denmark)

    Vinholt, Pernille Just; Nielsen, Christian; Söderström, Anna Cecilia

    2017-01-01

    Dabigatran is an oral anticoagulant and a reversible inhibitor of thrombin. Further, dabigatran might affect platelet function through a direct effect on platelet thrombin receptors. The aim was to investigate the effect of dabigatran on platelet activation and platelet aggregation. Healthy donor...

  4. Three different signal amplification strategies for the impedimetric sandwich detection of thrombin

    International Nuclear Information System (INIS)

    Ocaña, Cristina; Valle, Manel del

    2016-01-01

    In this work, we report a comparative study on three highly specific amplification strategies for the ultrasensitive detection of thrombin with the use of aptamer sandwich protocol. The protocol consisted on the use of a first thrombin aptamer immobilized on the electrode surface, the recognition of thrombin protein, and the reaction with a second biotinylated thrombin aptamer forming the sandwich. Through the exposed biotin end, three variants have been tested to amplify the electrochemical impedance signal. The strategies included (a) silver enhancement treatment, (b) gold enhancement treatment and (c) insoluble product produced by the combination of the enzyme horseradish peroxidase (HRP) and 3-amino-9-ethylcarbazole (AEC). The properties of the sensing surface were probed by electrochemical impedance measurements in the presence of the ferrocyanide/ferricyanide redox marker. Insoluble product strategy and silver enhancement treatment resulted in the lowest detection limit (0.3 pM), while gold enhancement method resulted in the highest reproducibility, 8.8% RSD at the pM thrombin concentration levels. Results of silver and gold enhancement treatment also permitted direct inspection by scanning electron microscopy (SEM). - Highlights: • Aptasensor to detect thrombin reaching the femtomolar level. • Biosensing protocol employs two thrombin aptamers in a sandwich capture scheme. • Use of second biotinylated aptamer allows many amplification and detection variants. • Precipitation reaction provides the highest signal amplification of ca. 3 times. • Double recognition event improves remarkably selectivity for thrombin detection.

  5. Structure of a retro-binding peptide inhibitor complexed with human alpha-thrombin.

    Science.gov (United States)

    Tabernero, L; Chang, C Y; Ohringer, S L; Lau, W F; Iwanowicz, E J; Han, W C; Wang, T C; Seiler, S M; Roberts, D G; Sack, J S

    1995-02-10

    The crystallographic structure of the ternary complex between human alpha-thrombin, hirugen and the peptidyl inhibitor Phe-alloThr-Phe-O-CH3, which is acylated at its N terminus with 4-guanidino butanoic acid (BMS-183507), has been determined at 2.6 A resolution. The structure reveals a unique "retro-binding" mode for this tripeptide active site inhibitor. The inhibitor binds with its alkyl-guanidine moiety in the primary specificity pocket and its two phenyl rings occupying the hydrophobic proximal and distal pockets of the thrombin active site. In this arrangement the backbone of the tripeptide forms a parallel beta-strand to the thrombin main-chain at the binding site. This is opposite to the orientation of the natural substrate, fibrinogen, and all the small active site-directed thrombin inhibitors whose bound structures have been previously reported. BMS-183507 is the first synthetic inhibitor proved to bind in a retro-binding fashion to thrombin, in a fashion similar to that of the N-terminal residues of the natural inhibitor hirudin. Furthermore, this new potent thrombin inhibitor (Ki = 17.2 nM) is selective for thrombin over other serine proteases tested and may be a template to be considered in designing hirudin-based thrombin inhibitors with interactions at the specificity pocket.

  6. Glycoprotein Ib activation by thrombin stimulates the energy metabolism in human platelets

    Science.gov (United States)

    Corona de la Peña, Norma; Gutiérrez-Aguilar, Manuel; Hernández-Reséndiz, Ileana; Marín-Hernández, Álvaro

    2017-01-01

    Thrombin-induced platelet activation requires substantial amounts of ATP. However, the specific contribution of each ATP-generating pathway i.e., oxidative phosphorylation (OxPhos) versus glycolysis and the biochemical mechanisms involved in the thrombin-induced activation of energy metabolism remain unclear. Here we report an integral analysis on the role of both energy pathways in human platelets activated by several agonists, and the signal transducing mechanisms associated with such activation. We found that thrombin, Trap-6, arachidonic acid, collagen, A23187, epinephrine and ADP significantly increased glycolytic flux (3–38 times vs. non-activated platelets) whereas ristocetin was ineffective. OxPhos (33 times) and mitochondrial transmembrane potential (88%) were increased only by thrombin. OxPhos was the main source of ATP in thrombin-activated platelets, whereas in platelets activated by any of the other agonists, glycolysis was the principal ATP supplier. In order to establish the biochemical mechanisms involved in the thrombin-induced OxPhos activation in platelets, several signaling pathways associated with mitochondrial activation were analyzed. Wortmannin and LY294002 (PI3K/Akt pathway inhibitors), ristocetin and heparin (GPIb inhibitors) as well as resveratrol, ATP (calcium-release inhibitors) and PP1 (Tyr-phosphorylation inhibitor) prevented the thrombin-induced platelet activation. These results suggest that thrombin activates OxPhos and glycolysis through GPIb-dependent signaling involving PI3K and Akt activation, calcium mobilization and protein phosphorylation. PMID:28817667

  7. A host-guest-recognition-based electrochemical aptasensor for thrombin detection.

    Science.gov (United States)

    Fan, Hao; Li, Hui; Wang, Qingjiang; He, Pingang; Fang, Yuzhi

    2012-05-15

    A sensitive electrochemical aptasensor for thrombin detection is presented based on the host-guest recognition technique. In this sensing protocol, a 15 based thrombin aptamer (ab. TBA) was dually labeled with a thiol at its 3' end and a 4-((4-(dimethylamino)phenyl)azo) benzoic acid (dabcyl) at its 5' end, respectively, which was previously immobilized on one Au electrode surface by AuS bond and used as the thrombin probe during the protein sensing procedure. One special electrochemical marker was prepared by modifying CdS nanoparticle with β-cyclodextrins (ab. CdS-CDs), which employed as electrochemical signal provider and would conjunct with the thrombin probe modified electrode through the host-guest recognition of CDs to dabcyl. In the absence of thrombin, the probe adopted linear structure to conjunct with CdS-CDs. In present of thrombin, the TBA bond with thrombin and transformed into its special G-quarter structure, which forced CdS-CDs into the solution. Therefore, the target-TBA binding event can be sensitively transduced via detecting the electrochemical oxidation current signal of Cd of CdS nanoparticles in the solution. Using this method, as low as 4.6 pM thrombin had been detected. Copyright © 2012 Elsevier B.V. All rights reserved.

  8. Adrenaline potentiates PI 3-kinase in platelets stimulated with thrombin and SFRLLN: role of secreted ADP.

    Science.gov (United States)

    Selheim, F; Frøyset, A K; Strand, I; Vassbotn, F S; Holmsen, H

    2000-11-17

    Adrenaline significantly potentiated late thrombin- and SFRLLN-induced PtdIns(3,4)P(2) production. Furthermore, the potentiating effect of adrenaline on thrombin-induced PtdIns(3, 4)P(2) production was independent on secreted ADP, whereas, the effect of adrenaline on SFRLLN-induced PtdIns(3,4)P(2) production was completely dependent of secreted ADP. However, the ADP-dependent accumulation of PtdIns(3,4)P(2) was not required for irreversible platelet aggregation induced by SFRLLN in the presence of adrenaline. It is concluded that adrenaline can replace secreted ADP to potentiate PtdIns(3,4)P(2) production in thrombin-stimulated but not in SFRLLN-stimulated platelets, thus demonstrating a qualitative difference between platelet stimulation by thrombin and the thrombin receptor activating peptide SFRLLN.

  9. Thrombin contributes to protective immunity in pneumonia-derived sepsis via fibrin polymerization and platelet-neutrophil interactions

    NARCIS (Netherlands)

    Claushuis, T. A. M.; de Stoppelaar, S. F.; Stroo, I.; Roelofs, J. J. T. H.; Ottenhoff, R.; van der Poll, T.; van't Veer, C.

    2017-01-01

    Essentials Immunity and coagulation are linked during sepsis but the role of thrombin is not fully elucidated. We investigated the effect of thrombin inhibition on murine Klebsiella pneumosepsis outcome. Thrombin is crucial for survival and limiting bacterial growth in pneumonia derived sepsis.

  10. Changes in thrombin generation in children after cardiac surgery and ex-vivo response to blood products and haemostatic agents

    DEFF Research Database (Denmark)

    Andreasen, Jo B; Ravn, Hanne B; Hvas, Anne-Mette

    2016-01-01

    surgery including cardiopulmonary bypass. Thrombin generation was analysed both in platelet-poor plasma and platelet-rich plasma. Analysis of the thrombin generation showed a significantly prolonged lag time (Pplatelet-poorandplatelet-richplasma ... thrombin generation significantly (all P rich plasma...

  11. Cation Coordination Alters the Conformation of a Thrombin-Binding G-Quadruplex DNA Aptamer That Affects Inhibition of Thrombin.

    Science.gov (United States)

    Zavyalova, Elena; Tagiltsev, Grigory; Reshetnikov, Roman; Arutyunyan, Alexander; Kopylov, Alexey

    2016-10-01

    Thrombin-binding aptamers are promising anticoagulants. HD1 is a monomolecular antiparallel G-quadruplex with two G-quartets linked by three loops. Aptamer-thrombin interactions are mediated with two TT-loops that bind thrombin exosite I. Several cations were shown to be coordinated inside the G-quadruplex, including K + , Na + , NH 4 + , Ba 2+ , and Sr 2+ ; on the contrary, Mn 2+ was coordinated in the grooves, outside the G-quadruplex. K + or Na + coordination provides aptamer functional activity. The effect of other cations on aptamer functional activity has not yet been described, because of a lack of relevant tests. Interactions between aptamer HD1 and a series of cations were studied. A previously developed enzymatic method was applied to evaluate aptamer inhibitory activity. The structure-function correlation was studied using the characterization of G-quadruplex conformation by circular dichroism spectroscopy. K + coordination provided the well-known high inhibitory activity of the aptamer, whereas Na + coordination supported low activity. Although NH 4 + coordination yielded a typical antiparallel G-quadruplex, no inhibitory activity was shown; a similar effect was observed for Ba 2+ and Sr 2+ coordination. Mn 2+ coordination destabilized the G-quadruplex that drastically diminished aptamer inhibitory activity. Therefore, G-quadruplex existence per se is insufficient for aptamer inhibitory activity. To elicit the nature of these effects, we thoroughly analyzed nuclear magnetic resonance (NMR) and X-ray data on the structure of the HD1 G-quadruplex with various cations. The most reasonable explanation is that cation coordination changes the conformation of TT-loops, affecting thrombin binding and inhibition. HD1 counterparts, aptamers 31-TBA and NU172, behaved similarly with some distinctions. In 31-TBA, an additional duplex module stabilized antiparallel G-quadruplex conformation at high concentrations of divalent cations; whereas in NU172, a different

  12. The pharmacological modulation of thrombin-induced cerebral thromboembolism in the rabbit.

    Science.gov (United States)

    May, G. R.; Paul, W.; Crook, P.; Butler, K. D.; Page, C. P.

    1992-01-01

    1. Intracarotid (i.c.) administration of thrombin induced a marked accumulation of 111indium-labelled platelets and 125I-labelled fibrinogen within the cranial vasculature of anaesthetized rabbits. 2. Thrombin (100 iu kg-1, i.c.) - induced platelet accumulation was completely abolished by pretreatment with desulphatohirudin (CGP 39393; 1 mg kg-1 i.c., 1 min prior to thrombin). Administration of CGP 39393 1 or 20 min after thrombin produced a significant reduction in platelet accumulation. 3. Intravenous (i.v.) administration of the platelet activating factor (PAF) receptor antagonist BN 52021 (10 mg kg-1) 5 min prior to thrombin (100 iu kg-1, i.c.) had no effect on platelet accumulation. 4. An inhibitor of NO biosynthesis, L-NG-nitro arginine methyl ester (L-NAME; 100 mg kg-1, i.c.), had no significant effect on the cranial platelet accumulation response to thrombin (10 iu kg-1, i.c.) when administered 5 min prior to thrombin. 5. Defibrotide (32 or 64 mg kg-1 bolus i.c. followed by 32 or 64 mg kg-1 h-1, i.c., infusion for 45 min) treatment begun 20 min after thrombin (100 iu kg-1, i.c.) did not significantly modify the cranial platelet accumulation response. 6. Cranial platelet accumulation induced by thrombin (100 iu kg-1, i.c.) was significantly reversed by the fibrinolytic drugs urokinase (20 iu kg-1, i.c., infusion for 45 min), anisoylated plasminogen streptokinase activator complex (APSAC) (200 micrograms kg-1, i.v. bolus) or recombinant tissue plasminogen activator (rt-PA; 100 micrograms kg-1, i.c. bolus followed by 20 micrograms kg-1 min-1, i.c., infusion for 45 min) administered 20 min after thrombin.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:1504722

  13. Thrombin-specific inactivation of endothelial cell derived plasminogen activator

    International Nuclear Information System (INIS)

    Highsmith, R.F.; Gallaher, M.J.

    1986-01-01

    Although thrombin (T) has diverse functions in the overall hemostatic mechanism, relatively little is known about its direct effect on components of the fibrinolytic enzyme system. The authors have investigated the interaction of T with plasminogen activators (PA) derived from bovine aortic endothelial cells (EC) in culture (2-5th passage, preconfluent monolayers). Varying concentrations of purified bovine or human thrombin were added to EC-conditioned media (CM). CM + T mixtures were assayed at various times for PA activity using purified plasminogen and a sensitive 125 I-fibrinogenolytic or caseinolytic assay. T (5 nM), but not plasmin or trypsin at equivalent concentrations, resulted in a time-dependent inhibition of the PA activity in CM. T had no effect on the PA activity of urokinase, streptokinase or preformed plasmin. The ability of T to inactivate the EC-derived PA was abolished by prior treatment of T with active site-directed reagents. SDS-PAGE and zymography with copolymerized fibrinogen and plasminogen revealed further specificity in that only one of the multiple-molecular weight forms of PA present in EC-CM was inactivated by T. The authors conclude that in a highly specific fashion, T inactivates the predominant PA present in EC-CM by limited proteolysis. Thus, another potentially important function of T is suggested which may have particular significance in the temporal regulation of coagulation and fibrinolysis at the blood-endothelium interface

  14. Thrombin-specific inactivation of endothelial cell derived plasminogen activator

    Energy Technology Data Exchange (ETDEWEB)

    Highsmith, R.F.; Gallaher, M.J.

    1986-03-05

    Although thrombin (T) has diverse functions in the overall hemostatic mechanism, relatively little is known about its direct effect on components of the fibrinolytic enzyme system. The authors have investigated the interaction of T with plasminogen activators (PA) derived from bovine aortic endothelial cells (EC) in culture (2-5th passage, preconfluent monolayers). Varying concentrations of purified bovine or human thrombin were added to EC-conditioned media (CM). CM + T mixtures were assayed at various times for PA activity using purified plasminogen and a sensitive /sup 125/I-fibrinogenolytic or caseinolytic assay. T (5 nM), but not plasmin or trypsin at equivalent concentrations, resulted in a time-dependent inhibition of the PA activity in CM. T had no effect on the PA activity of urokinase, streptokinase or preformed plasmin. The ability of T to inactivate the EC-derived PA was abolished by prior treatment of T with active site-directed reagents. SDS-PAGE and zymography with copolymerized fibrinogen and plasminogen revealed further specificity in that only one of the multiple-molecular weight forms of PA present in EC-CM was inactivated by T. The authors conclude that in a highly specific fashion, T inactivates the predominant PA present in EC-CM by limited proteolysis. Thus, another potentially important function of T is suggested which may have particular significance in the temporal regulation of coagulation and fibrinolysis at the blood-endothelium interface.

  15. Contact system activation and high thrombin generation in hyperthyroidism.

    Science.gov (United States)

    Kim, Namhee; Gu, Ja-Yoon; Yoo, Hyun Ju; Han, Se Eun; Kim, Young Il; Nam-Goong, Il Sung; Kim, Eun Sook; Kim, Hyun Kyung

    2017-05-01

    Hyperthyroidism is associated with increased thrombotic risk. As contact system activation through formation of neutrophil extracellular traps (NET) has emerged as an important trigger of thrombosis, we hypothesized that the contact system is activated along with active NET formation in hyperthyroidism and that their markers correlate with disease severity. In 61 patients with hyperthyroidism and 40 normal controls, the levels of coagulation factors (fibrinogen, and factor VII, VIII, IX, XI and XII), D-dimer, thrombin generation assay (TGA) markers, NET formation markers (histone-DNA complex, double-stranded DNA and neutrophil elastase) and contact system markers (activated factor XII (XIIa), high-molecular-weight kininogen (HMWK), prekallikrein and bradykinin) were measured. Patients with hyperthyroidism showed higher levels of fibrinogen (median (interquartile range), 315 (280-344) vs 262 (223-300), P  = 0.001), D-dimer (103.8 (64.8-151.5) vs 50.7 (37.4-76.0), P  hyperthyroidism's contribution to coagulation and contact system activation. Free T4 was significantly correlated with factors VIII and IX, D-dimer, double-stranded DNA and bradykinin. This study demonstrated that contact system activation and abundant NET formation occurred in the high thrombin generation state in hyperthyroidism and were correlated with free T4 level. © 2017 European Society of Endocrinology.

  16. Cross-calibration of interferometric SAR data

    DEFF Research Database (Denmark)

    Dall, Jørgen

    2003-01-01

    Generation of digital elevation models from interferometric synthetic aperture radar (SAR) data is a well established technique. Achieving a high geometric fidelity calls for a calibration accounting for inaccurate navigation data and system parameters as well as system imperfections. Fully...... automated calibration techniques are preferable, especially for operational mapping. The author presents one such technique, called cross-calibration. Though developed for single-pass interferometry, it may be applicable to multi-pass interferometry, too. Cross-calibration requires stability during mapping...... ground control point is often needed. The paper presents the principles and mathematics of the cross-calibration technique and illustrates its successful application to EMISAR data....

  17. Multiple-Objective Stepwise Calibration Using Luca

    Science.gov (United States)

    Hay, Lauren E.; Umemoto, Makiko

    2007-01-01

    This report documents Luca (Let us calibrate), a multiple-objective, stepwise, automated procedure for hydrologic model calibration and the associated graphical user interface (GUI). Luca is a wizard-style user-friendly GUI that provides an easy systematic way of building and executing a calibration procedure. The calibration procedure uses the Shuffled Complex Evolution global search algorithm to calibrate any model compiled with the U.S. Geological Survey's Modular Modeling System. This process assures that intermediate and final states of the model are simulated consistently with measured values.

  18. ARTIP: Automated Radio Telescope Image Processing Pipeline

    Science.gov (United States)

    Sharma, Ravi; Gyanchandani, Dolly; Kulkarni, Sarang; Gupta, Neeraj; Pathak, Vineet; Pande, Arti; Joshi, Unmesh

    2018-02-01

    The Automated Radio Telescope Image Processing Pipeline (ARTIP) automates the entire process of flagging, calibrating, and imaging for radio-interferometric data. ARTIP starts with raw data, i.e. a measurement set and goes through multiple stages, such as flux calibration, bandpass calibration, phase calibration, and imaging to generate continuum and spectral line images. Each stage can also be run independently. The pipeline provides continuous feedback to the user through various messages, charts and logs. It is written using standard python libraries and the CASA package. The pipeline can deal with datasets with multiple spectral windows and also multiple target sources which may have arbitrary combinations of flux/bandpass/phase calibrators.

  19. Macrophage Migration Inhibitory Factor-Induced Autophagy Contributes to Thrombin-Triggered Endothelial Hyperpermeability in Sepsis.

    Science.gov (United States)

    Chao, Chiao-Hsuan; Chen, Hong-Ru; Chuang, Yung-Chun; Yeh, Trai-Ming

    2018-07-01

    Vascular leakage contributes to the high morbidity and mortality associated with sepsis. Exposure of the endothelium to inflammatory mediators, such as thrombin and cytokines, during sepsis leads to hyperpermeability. We recently observed that autophagy, a cellular process for protein turnover, is involved in macrophage migration inhibitory factor (MIF)-induced endothelial hyperpermeability. Even though it is known that thrombin induces endothelial cells to secrete MIF and to increase vascular permeability, the possible role of autophagy in this process is unknown. In this study, we proposed and tested the hypothesis that MIF-induced autophagy plays an important role in thrombin-induced endothelial hyperpermeability. We evaluated the effects of thrombin on endothelial permeability, autophagy induction, and MIF secretion in vitro using the human microvascular endothelial cell line-1 and human umbilical vein endothelial cells. Several mechanisms/read outs of endothelial permeability and autophagy formation were examined. We observed that blocking autophagy attenuated thrombin-induced endothelial hyperpermeability. Furthermore, thrombin-induced MIF secretion was involved in this process because MIF inhibition reduced thrombin-induced autophagy and hyperpermeability. Finally, we showed that blocking MIF or autophagy effectively alleviated vascular leakage and mortality in endotoxemic mice. Thus, MIF-induced autophagy may represent a common mechanism causing vascular leakage in sepsis.

  20. Thermodynamic, Anticoagulant, and Antiproliferative Properties of Thrombin Binding Aptamer Containing Novel UNA Derivative

    DEFF Research Database (Denmark)

    Kotkowiak, Weronika; Lisowiec-Wachnicka, Jolanta; Grynda, Jakub

    2018-01-01

    Thrombin is a serine protease that plays a crucial role in hemostasis, fibrinolysis, cell proliferation, and migration. Thrombin binding aptamer (TBA) is able to inhibit the activity of thrombin molecule via binding to its exosite I. This 15-nt DNA oligonucleotide forms an intramolecular, antipar......Thrombin is a serine protease that plays a crucial role in hemostasis, fibrinolysis, cell proliferation, and migration. Thrombin binding aptamer (TBA) is able to inhibit the activity of thrombin molecule via binding to its exosite I. This 15-nt DNA oligonucleotide forms an intramolecular......, antiparallel G-quadruplex structure with a chair-like conformation. In this paper, we report on our investigations on the influence of certain modified nucleotide residues on thermodynamic stability, folding topology, and biological properties of TBA variants. In particular, the effect of single incorporation......-quadruplex thermodynamic and biological stability, and that the effect is strongly position dependent. Interestingly, TBA variants containing the modified nucleotide residues are characterized by unchanged folding topology. Thrombin time assay revealed that incorporation of certain UNA residues may improve G...

  1. Protamine sulfate down-regulates thrombin generation by inhibiting factor V activation.

    LENUS (Irish Health Repository)

    Ni Ainle, Fionnuala

    2009-08-20

    Protamine sulfate is a positively charged polypeptide widely used to reverse heparin-induced anticoagulation. Paradoxically, prospective randomized trials have shown that protamine administration for heparin neutralization is associated with increased bleeding, particularly after cardiothoracic surgery with cardiopulmonary bypass. The molecular mechanism(s) through which protamine mediates this anticoagulant effect has not been defined. In vivo administration of pharmacologic doses of protamine to BALB\\/c mice significantly reduced plasma thrombin generation and prolonged tail-bleeding time (from 120 to 199 seconds). Similarly, in pooled normal human plasma, protamine caused significant dose-dependent prolongations of both prothrombin time and activated partial thromboplastin time. Protamine also markedly attenuated tissue factor-initiated thrombin generation in human plasma, causing a significant decrease in endogenous thrombin potential (41% +\\/- 7%). As expected, low-dose protamine effectively reversed the anticoagulant activity of unfractionated heparin in plasma. However, elevated protamine concentrations were associated with progressive dose-dependent reduction in thrombin generation. To assess the mechanism by which protamine mediates down-regulation of thrombin generation, the effect of protamine on factor V activation was assessed. Protamine was found to significantly reduce the rate of factor V activation by both thrombin and factor Xa. Protamine mediates its anticoagulant activity in plasma by down-regulation of thrombin generation via a novel mechanism, specifically inhibition of factor V activation.

  2. Thrombin-inhibiting nanoparticles rapidly constitute versatile and detectable anticlotting surfaces

    Science.gov (United States)

    Wheatley Myerson, Jacob; He, Li; Allen, John Stacy; Williams, Todd; Lanza, Gregory; Tollefsen, Douglas; Caruthers, Shelton; Wickline, Samuel

    2014-09-01

    Restoring an antithrombotic surface to suppress ongoing thrombosis is an appealing strategy for treatment of acute cardiovascular disorders such as erosion of atherosclerotic plaque. An antithrombotic surface would present an alternative to systemic anticoagulation with attendant risks of bleeding. We have designed thrombin-targeted nanoparticles (NPs) that bind to sites of active clotting to extinguish local thrombin activity and inhibit platelet deposition while exhibiting only transient systemic anticoagulant effects. Perfluorocarbon nanoparticles (PFC NP) were functionalized with thrombin inhibitors (either D-phenylalanyl-L-prolyl-L-arginyl-chloromethyl ketone or bivalirudin) by covalent attachment of more than 15 000 inhibitors to each PFC NP. Fibrinopeptide A (FPA) ELISA demonstrated that thrombin-inhibiting NPs prevented cleavage of fibrinogen by both free and clot-bound thrombin. Magnetic resonance imaging (MRI) confirmed that a layer of thrombin-inhibiting NPs prevented growth of clots in vitro. Thrombin-inhibiting NPs were administered in vivo to C57BL6 mice subjected to laser injury of the carotid artery. NPs significantly delayed thrombotic occlusion of the artery, whereas an equivalent bolus of free inhibitor was ineffective. For thrombin-inhibiting NPs, only a short-lived (˜10 min) systemic effect on bleeding time was observed, despite prolonged clot inhibition. Imaging and quantification of in vivo antithrombotic NP layers was demonstrated by MRI of the PFC NP. 19F MRI confirmed colocalization of particles with arterial thrombi, and quantitative 19F spectroscopy demonstrated specific binding and retention of thrombin-inhibiting NPs in injured arteries. The ability to rapidly form and image a new antithrombotic surface in acute vascular syndromes while minimizing risks of bleeding would permit a safer method of passivating active lesions than current systemic anticoagulant regimes.

  3. Thrombin-inhibiting nanoparticles rapidly constitute versatile and detectable anticlotting surfaces

    International Nuclear Information System (INIS)

    Myerson, Jacob Wheatley; Lanza, Gregory; Caruthers, Shelton; Wickline, Samuel; He, Li; Allen, John Stacy; Williams, Todd; Tollefsen, Douglas

    2014-01-01

    Restoring an antithrombotic surface to suppress ongoing thrombosis is an appealing strategy for treatment of acute cardiovascular disorders such as erosion of atherosclerotic plaque. An antithrombotic surface would present an alternative to systemic anticoagulation with attendant risks of bleeding. We have designed thrombin-targeted nanoparticles (NPs) that bind to sites of active clotting to extinguish local thrombin activity and inhibit platelet deposition while exhibiting only transient systemic anticoagulant effects. Perfluorocarbon nanoparticles (PFC NP) were functionalized with thrombin inhibitors (either D-phenylalanyl-L-prolyl-L-arginyl-chloromethyl ketone or bivalirudin) by covalent attachment of more than 15 000 inhibitors to each PFC NP. Fibrinopeptide A (FPA) ELISA demonstrated that thrombin-inhibiting NPs prevented cleavage of fibrinogen by both free and clot-bound thrombin. Magnetic resonance imaging (MRI) confirmed that a layer of thrombin-inhibiting NPs prevented growth of clots in vitro. Thrombin-inhibiting NPs were administered in vivo to C57BL6 mice subjected to laser injury of the carotid artery. NPs significantly delayed thrombotic occlusion of the artery, whereas an equivalent bolus of free inhibitor was ineffective. For thrombin-inhibiting NPs, only a short-lived (∼10 min) systemic effect on bleeding time was observed, despite prolonged clot inhibition. Imaging and quantification of in vivo antithrombotic NP layers was demonstrated by MRI of the PFC NP. 19 F MRI confirmed colocalization of particles with arterial thrombi, and quantitative 19 F spectroscopy demonstrated specific binding and retention of thrombin-inhibiting NPs in injured arteries. The ability to rapidly form and image a new antithrombotic surface in acute vascular syndromes while minimizing risks of bleeding would permit a safer method of passivating active lesions than current systemic anticoagulant regimes. (paper)

  4. Synergism between thrombin and adrenaline (epinephrine) in human platelets. Marked potentiation of inositol phospholipid metabolism.

    Science.gov (United States)

    Steen, V M; Tysnes, O B; Holmsen, H

    1988-01-01

    We have studied synergism between adrenaline (epinephrine) and low concentrations of thrombin in gel-filtered human platelets prelabelled with [32P]Pi. Suspensions of platelets, which did not contain added fibrinogen, were incubated at 37 degrees C to measure changes in the levels of 32P-labelled phosphatidylinositol 4,5-bisphosphate (PIP2), phosphatidylinositol 4-phosphate (PIP) and phosphatidate (PA), aggregation and dense-granule secretion after stimulation. Adrenaline alone (3.5-4.0 microM) did not cause a change in any parameter (phosphoinositide metabolism, aggregation and dense-granule secretion), but markedly enhanced the thrombin-induced responses over a narrow range of thrombin concentrations (0.03-0.08 units/ml). The thrombin-induced hydrolysis of inositol phospholipids by phospholipase C, which was measured as the formation of [32P]PA, was potentiated by adrenaline, as was the increase in the levels of [32P]PIP2 and [32P]PIP. The presence of adrenaline caused a shift to the left for the thrombin-induced changes in the phosphoinositide metabolism, without affecting the maximal levels of 32P-labelled compounds obtained. A similar shift by adrenaline in the dose-response relationship was previously demonstrated for thrombin-induced aggregation and dense-granule secretion. Also, the narrow range of concentrations of thrombin over which adrenaline potentiates thrombin-induced platelet responses is the same for changes in phosphoinositide metabolism and physiological responses (aggregation and dense-granule secretion). Our observations clearly indicate that adrenaline directly or indirectly influences thrombin-induced changes in phosphoinositide metabolism. PMID:2845924

  5. Loop Electrostatics Asymmetry Modulates the Preexisting Conformational Equilibrium in Thrombin.

    Science.gov (United States)

    Pozzi, Nicola; Zerbetto, Mirco; Acquasaliente, Laura; Tescari, Simone; Frezzato, Diego; Polimeno, Antonino; Gohara, David W; Di Cera, Enrico; De Filippis, Vincenzo

    2016-07-19

    Thrombin exists as an ensemble of active (E) and inactive (E*) conformations that differ in their accessibility to the active site. Here we show that redistribution of the E*-E equilibrium can be achieved by perturbing the electrostatic properties of the enzyme. Removal of the negative charge of the catalytic Asp102 or Asp189 in the primary specificity site destabilizes the E form and causes a shift in the 215-217 segment that compromises substrate entrance. Solution studies and existing structures of D102N document stabilization of the E* form. A new high-resolution structure of D189A also reveals the mutant in the collapsed E* form. These findings establish a new paradigm for the control of the E*-E equilibrium in the trypsin fold.

  6. Behaviour of homologous 125I fibrinogen after thrombin and ancrod infusion in rabbits

    International Nuclear Information System (INIS)

    Setter, R.

    1977-01-01

    The behaviour of radioactively labelled fibrinogen after infusion of thrombin or ancrod is investigated. Common factors and differences in the behaviour of fibrinogen after infusion of these two enzymes, which act proteolytically on the fibrinogen, are dealt with. Rabbits received an i.v. injection of homologous 125 I-fibrinogen 3 days before ancrod or thrombin infusion. On the day of the experiments, one group of animals received an ancrod infusion (1.5 U/kg body weight for 30 minutes), the other a thrombin infusion (600 U/kg body weight for 60 minutes). Intravenous ancrod and thrombin infusions lowered the fibrinogen level to 30% or 50% of the initial value due to intravascular coagulation. About 50% of the 125 I fibrinogen was transformed after ancrod exposure into a non-coagulating fraction of fibrinogen derivatives which produces no fibrinolytic decomposition products. (orig./AJ) [de

  7. Comparing thrombin generation in patients with hemophilia A and patients on vitamin K antagonists

    NARCIS (Netherlands)

    de Koning, M L Y; Fischer, K; de Laat, B; Huisman, A; Ninivaggi, M; Schutgens, R E G

    Essentials: It is unknown if hemophilia patients with atrial fibrillation need anticoagulation. Endogenous thrombin potentials (ETP) in hemophilia patients and patients on coumarins were compared. Severe hemophilia patients had comparable ETP to therapeutic international normalized ratio (INR). In

  8. Active but inoperable thrombin is accumulated in a plasma protein layer surrounding Streptococcus pyogenes

    NARCIS (Netherlands)

    Naudin, Clément; Hurley, Sinead M.; Malmström, Erik; Plug, Tom; Shannon, Oonagh; Meijers, Joost C. M.; Mörgelin, Matthias; Björck, Lars; Herwald, Heiko

    2015-01-01

    Activation of thrombin is a critical determinant in many physiological and pathological processes including haemostasis and inflammation. Under physiological conditions many of these functions are involved in wound healing or eradication of an invading pathogen. However, when activated systemically,

  9. Thrombin stimulates albumin transcytosis in lung microvascular endothelial cells via activation of acid sphingomyelinase.

    Science.gov (United States)

    Kuebler, Wolfgang M; Wittenberg, Claudia; Lee, Warren L; Reppien, Eike; Goldenberg, Neil M; Lindner, Karsten; Gao, Yizhuo; Winoto-Morbach, Supandi; Drab, Marek; Mühlfeld, Christian; Dombrowsky, Heike; Ochs, Matthias; Schütze, Stefan; Uhlig, Stefan

    2016-04-15

    Transcellular albumin transport occurs via caveolae that are abundant in lung microvascular endothelial cells. Stimulation of albumin transcytosis by proinflammatory mediators may contribute to alveolar protein leak in lung injury, yet the regulation of albumin transport and its underlying molecular mechanisms are so far incompletely understood. Here we tested the hypothesis that thrombin may stimulate transcellular albumin transport across lung microvascular endothelial cells in an acid-sphingomyelinase dependent manner. Thrombin increased the transport of fluorescently labeled albumin across confluent human lung microvascular endothelial cell (HMVEC-L) monolayers to an extent that markedly exceeds the rate of passive diffusion. Thrombin activated acid sphingomyelinase (ASM) and increased ceramide production in HMVEC-L, but not in bovine pulmonary artery cells, which showed little albumin transport in response to thrombin. Thrombin increased total caveolin-1 (cav-1) content in both whole cell lysates and lipid rafts from HMVEC-L, and this effect was blocked by inhibition of ASM or de novo protein biosynthesis. Thrombin-induced uptake of albumin into lung microvascular endothelial cells was confirmed in isolated-perfused lungs by real-time fluorescence imaging and electron microscopy of gold-labeled albumin. Inhibition of ASM attenuated thrombin-induced albumin transport both in confluent HMVEC-L and in intact lungs, whereas HMVEC-L treatment with exogenous ASM increased albumin transport and enriched lipid rafts in cav-1. Our findings indicate that thrombin stimulates transcellular albumin transport in an acid sphingomyelinase-dependent manner by inducing de novo synthesis of cav-1 and its recruitment to membrane lipid rafts. Copyright © 2016 the American Physiological Society.

  10. Synergistic action between inhibition of P2Y12/P2Y1 and P2Y12/thrombin in ADP- and thrombin-induced human platelet activation

    Science.gov (United States)

    Nylander, Sven; Mattsson, Christer; Ramström, Sofia; Lindahl, Tomas L

    2004-01-01

    The objective of this study was to investigate if there is a synergistic effect of a combination of P2Y12 and P2Y1 inhibition and P2Y12 and thrombin inhibition, on ADP- and thrombin-induced platelet activation, respectively. The rationale being that these combinations will cause a concurrent inhibition of both Gαq and Gαi signalling.Blood from healthy volunteers was preincubated with AR-C69931MX, a reversible P2Y12 antagonist; MRS2179, a reversible P2Y1 antagonist; or melagatran, a direct reversible thrombin inhibitor; alone or in various combinations prior to activation with ADP or thrombin. Platelet function in whole blood was assessed by flow cytometry using the antibody PAC-1 to estimate the expression of active αIIbβ3 (the fibrinogen receptor GPIIb/IIIa). A synergistic effect was evaluated by comparing the concentrations in the different combinations with those of corresponding equipotent concentrations of each single inhibitor alone. The equipotent single concentrations were experimentally obtained from concentration response curves performed in parallel.A synergistic effect regarding inhibition of ADP-induced platelet activation (10 μM) was obtained with different combinations of AR-C69931MX and MRS2179.Inhibition of thrombin-induced platelet activation (2 nM) with combinations of AR-C69931MX and the thrombin inhibitor melagatran did also result in a strong synergistic effect.To our knowledge, this is the first time that data supporting a synergistic effect has been published for the inhibitor combinations described.Whether this synergistic effect in vitro also results in an improved antithrombotic effect in vivo with or without an increased risk of bleeding remains to be studied in well-conducted clinical studies. PMID:15265806

  11. Thrombin Production and Human Neutrophil Elastase Sequestration by Modified Cellulosic Dressings and Their Electrokinetic Analysis

    Directory of Open Access Journals (Sweden)

    Nicolette Prevost

    2011-12-01

    Full Text Available Wound healing is a complex series of biochemical and cellular events. Optimally, functional material design addresses the overlapping acute and inflammatory stages of wound healing based on molecular, cellular, and bio-compatibility issues. In this paper the issues addressed are uncontrolled hemostasis and inflammation which can interfere with the orderly flow of wound healing. In this regard, we review the serine proteases thrombin and elastase relative to dressing functionality that improves wound healing and examine the effects of charge in cotton/cellulosic dressing design on thrombin production and elastase sequestration (uptake by the wound dressing. Thrombin is central to the initiation and propagation of coagulation, and elastase is released from neutrophils that can function detrimentally in a stalled inflammatory phase characteristic of chronic wounds. Electrokinetic fiber surface properties of the biomaterials of this study were determined to correlate material charge and polarity with function relative to thrombin production and elastase sequestration. Human neutrophil elastase sequestration was assessed with an assay representative of chronic wound concentration with cotton gauze cross-linked with three types of polycarboxylic acids and one phosphorylation finish; thrombin production, which was assessed in a plasma-based assay via a fluorogenic peptide substrate, was determined for cotton, cotton-grafted chitosan, chitosan, rayon/polyester, and two kaolin-treated materials including a commercial hemorrhage control dressing (QuickClot Combat Gauze. A correlation in thrombin production to zeta potential was found. Two polycarboxylic acid cross linked and a phosphorylated cotton dressing gave high elastase sequestration.

  12. The Phosphatase Inhibitor Calyculin-A Impairs Clot Retraction, Platelet Activation, and Thrombin Generation

    Directory of Open Access Journals (Sweden)

    Renáta Hudák

    2017-01-01

    Full Text Available The aim of this study was to investigate the effect of the serine/threonine protein phosphatase inhibitor, calyculin-A (CLA, on clot formation and on the procoagulant activity of human platelets. Platelet-rich plasma (PRP samples were preincubated with buffer or CLA and subsequently platelets were activated by the protease-activated receptor 1 (PAR-1 activator, thrombin receptor activating peptide (TRAP. Clot retraction was detected by observing clot morphology up to 1 hour, phosphatidylserine- (PS- expression was studied by flow cytometry, and thrombin generation was measured by a fluorimetric assay. For the intracellular Ca2+ assay, platelets were loaded with calcium-indicator dyes and the measurements were carried out using a ratiometric method with real-time confocal microscopy. CLA preincubation inhibited clot retraction, PS-expression, and thrombin formation. TRAP activation elicited Ca2+ response and PS-expression in a subset of platelets. The activated PRP displayed significantly faster and enhanced thrombin generation compared to nonactivated samples. CLA pretreatment abrogated PS-exposure and clot retraction also in TRAP-activated samples. As a consequence of the inhibitory effect on calcium elevation and PS-expression, CLA significantly downregulated thrombin generation in PRP. Our results show that CLA pretreatment may be a useful tool to investigate platelet activation mechanisms that contribute to clot formation and thrombin generation.

  13. Synergy in thrombin-graphene sponge for improved hemostatic efficacy and facile utilization.

    Science.gov (United States)

    Li, Guofeng; Quan, Kecheng; Xu, CongCong; Deng, Bo; Wang, Xing

    2018-01-01

    Composites are attractive for its potential synergistic effects that can result in high-performance, but the synergy depends on subtle design. In this study, a hemostatic composite, a thrombin/cross-linked graphene sponge (TCGS), was developed through a facile gradient composite strategy. The porous structure of the CGS assures that the thrombin is stably embedded in the TCGS, avoiding a burst release but maintaining its bioactivity. In the synergy between proper thrombin stimulation and the fast absorption of the sponge, TCGS exhibits outstanding hemostatic performance, ultrafast bleeding cessation, within 100s, which is superior to both CGS and equal amounts of native thrombin. Lower or excessive thrombin dosages prolong the bleeding time. The study revealed that the balance between plasma absorption and thrombin stimulation at the interface is critical for improving hemostatic efficacy. TCGS is also highlighted for its biosafety and stability, even after 6 months of storage in environment. This potentially ultra-long shelf life is conducive to its practical applications. Therefore, TCGS not only provides a new strategy for developing a hemostatic composite but also provides a new method and understanding for the design of hemostatic materials. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Thrombin-Activatable Microbubbles as Potential Ultrasound Contrast Agents for the Detection of Acute Thrombosis.

    Science.gov (United States)

    Lux, Jacques; Vezeridis, Alexander M; Hoyt, Kenneth; Adams, Stephen R; Armstrong, Amanda M; Sirsi, Shashank R; Mattrey, Robert F

    2017-11-01

    Acute deep vein thrombosis (DVT) is the formation of a blood clot in the deep veins of the body that can lead to fatal pulmonary embolism. Acute DVT is difficult to distinguish from chronic DVT by ultrasound (US), the imaging modality of choice, and is therefore treated aggressively with anticoagulants, which can lead to internal bleeding. Here we demonstrate that conjugating perfluorobutane-filled (PFB-filled) microbubbles (MBs) with thrombin-sensitive activatable cell-penetrating peptides (ACPPs) could lead to the development of contrast agents that detect acute thrombosis with US imaging. Successful conjugation of ACPP to PFB-filled MBs was confirmed by fluorescence microscopy and flow cytometry. Fluorescein-labeled ACPP was used to evaluate the efficiency of thrombin-triggered cleavage by measuring the mean fluorescence intensity of ACPP-labeled MBs (ACPP-MBs) before and after incubation at 37 °C with thrombin. Lastly, control MBs and ACPP-MBs were infused through a tube containing a clot, and US contrast enhancement was measured with or without the presence of a thrombin inhibitor after washing the clot with saline. With thrombin activity, 91.7 ± 14.2% of the signal was retained after ACPP-MB infusion and washing, whereas only 16.7 ± 4% of the signal was retained when infusing ACPP-MBs in the presence of hirudin, a potent thrombin inhibitor.

  15. Investigation of the heparin-thrombin interaction by dynamic force spectroscopy.

    Science.gov (United States)

    Wang, Congzhou; Jin, Yingzi; Desai, Umesh R; Yadavalli, Vamsi K

    2015-06-01

    The interaction between heparin and thrombin is a vital step in the blood (anti)coagulation process. Unraveling the molecular basis of the interactions is therefore extremely important in understanding the mechanisms of this complex biological process. In this study, we use a combination of an efficient thiolation chemistry of heparin, a self-assembled monolayer-based single molecule platform, and a dynamic force spectroscopy to provide new insights into the heparin-thrombin interaction from an energy viewpoint at the molecular scale. Well-separated single molecules of heparin covalently attached to mixed self-assembled monolayers are demonstrated, whereby interaction forces with thrombin can be measured via atomic force microscopy-based spectroscopy. Further these interactions are studied at different loading rates and salt concentrations to directly obtain kinetic parameters. An increase in the loading rate shows a higher interaction force between the heparin and thrombin, which can be directly linked to the kinetic dissociation rate constant (koff). The stability of the heparin/thrombin complex decreased with increasing NaCl concentration such that the off-rate was found to be driven primarily by non-ionic forces. These results contribute to understanding the role of specific and nonspecific forces that drive heparin-thrombin interactions under applied force or flow conditions. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. Instrumentation calibration

    International Nuclear Information System (INIS)

    Mack, D.A.

    1976-08-01

    Procedures for the calibration of different types of laboratory equipment are described. Provisions for maintaining the integrity of reference and working standards traceable back to a national standard are discussed. Methods of validation and certification methods are included. An appendix lists available publications and services of national standardizing agencies

  17. Presence of plasma proteins facilitates the uptake of 125I-thrombin by the rabbit thoracic aorta endothelium in vitro

    International Nuclear Information System (INIS)

    Hatton, M.W.; Moar, S.L.

    1986-01-01

    Various purified proteins, protein derivatives and two polysaccharides were added individually to a physiological medium in order to effect uptake of 125 I-thrombin by the rabbit aorta endothelium. Over a wide range of concentration (0.004-40 mg/ml), the presence of either purified rabbit or bovine albumin during thrombin uptake encouraged an increase (70-110%) in 125 I-thrombin binding by the endothelium and subendothelium compared to uptake by aorta segments in the absence of added protein. Pretreatment of aorta segments with albumin before incubation with 125 I-thrombin in the absence of albumin did not encourage thrombin uptake to the same extent as having 125 I-thrombin and albumin together. Purified human transferrin, rabbit IgG, chicken ovalbumin or denatured bovine casein could replace albumin to produce a similar enhancement of thrombin uptake. Replacing active concentrations of albumin by either reduced-carboxymethylated albumin, defatted albumin, plasmin-treated or thermolysin-treated albumin also caused an increase (50-130%) in thrombin binding, whereas replacement by acid-hydrolysed albumin or with polyglutamic acid was either ineffective or even inhibitory. Lysine-modified or arginine-modified albumins caused a small enhancement (14-32%) and no enhancement of thrombin uptake, respectively. Dextran, at low concentration (0.04-0.4 mg/ml) did not influence thrombin uptake, and at higher concentration (4-40 mg/ml) caused a decrease in uptake by both the endothelium and subendothelial layers. Low concentration of dextran sulphate inhibited thrombin uptake to 20-30% of control values. These data express the importance of accompanying protein in the response of the vascular endothelium during binding of thrombin. The possibility that other protein-cell interactions may be similarly influenced by macromolecular solutes is also discussed

  18. Lipopolysaccharide interactions of C-terminal peptides from human thrombin.

    Science.gov (United States)

    Singh, Shalini; Kalle, Martina; Papareddy, Praveen; Schmidtchen, Artur; Malmsten, Martin

    2013-05-13

    Interactions with bacterial lipopolysaccharide (LPS), both in aqueous solution and in lipid membranes, were investigated for a series of amphiphilic peptides derived from the C-terminal region of human thrombin, using ellipsometry, dual polarization interferometry, fluorescence spectroscopy, circular dichroism (CD), dynamic light scattering, and z-potential measurements. The ability of these peptides to block endotoxic effects caused by LPS, monitored through NO production in macrophages, was compared to peptide binding to LPS and its endotoxic component lipid A, and to size, charge, and secondary structure of peptide/LPS complexes. While the antiendotoxic peptide GKY25 (GKYGFYTHVFRLKKWIQKVIDQFGE) displayed significant binding to both LPS and lipid A, so did two control peptides with either selected D-amino acid substitutions or with maintained composition but scrambled sequence, both displaying strongly attenuated antiendotoxic effects. Hence, the extent of LPS or lipid A binding is not the sole discriminant for the antiendotoxic effect of these peptides. In contrast, helix formation in peptide/LPS complexes correlates to the antiendotoxic effect of these peptides and is potentially linked to this functionality. Preferential binding to LPS over lipid membrane was furthermore demonstrated for these peptides and preferential binding to the lipid A moiety within LPS inferred.

  19. Thrombin-Binding Aptamer Quadruplex Formation: AFM and Voltammetric Characterization

    Directory of Open Access Journals (Sweden)

    Victor Constantin Diculescu

    2010-01-01

    Full Text Available The adsorption and the redox behaviour of thrombin-binding aptamer (TBA and extended TBA (eTBA were studied using atomic force microscopy and voltammetry at highly oriented pyrolytic graphite and glassy carbon. The different adsorption patterns and degree of surface coverage were correlated with the sequence base composition, presence/absence of K+, and voltammetric behaviour of TBA and eTBA. In the presence of K+, only a few single-stranded sequences present adsorption, while the majority of the molecules forms stable and rigid quadruplexes with no adsorption. Both TBA and eTBA are oxidized and the only anodic peak corresponds to guanine oxidation. Upon addition of K+ ions, TBA and eTBA fold into a quadruplex, causing the decrease of guanine oxidation peak and occurrence of a new peak at a higher potential due to the oxidation of G-quartets. The higher oxidation potential of G-quartets is due to the greater difficulty of electron transfer from the inside of the quadruplex to the electrode surface than electron transfer from the more flexible single strands.

  20. Thrombin activatable fibrinolysis inhibitor (TAFI in cord blood.

    Directory of Open Access Journals (Sweden)

    Krzysztof Góralczyk

    2007-03-01

    Full Text Available Thrombin activatable fibrinolysis inhibitor (TAFI is a plasma zymogene (procarboxypeptidase B which can decrease fibrinolysis and thus act as a haemostatic factor. TAFI is now extensively studied in many complications as well as in physiological and complicated pregnancy. The question we posed in the present study was whether TAFI antigen is present in cord blood plasma. The study group consisted of 38 parturient women, 26 primiparous and 12 multiparous with normal course of pregnancy and delivery. The cord blood was sampled from the cord vein, and the mother's blood from the antecubital vein. 3.2% sodium citrate was used as an anticoagulant. TAFIa/ai antigen was measured by ELISA method. TAFIa/ai antigen was identified in all samples of cord blood plasma. Its level was 91.50 ng/ml (range: 71.76 - 160.77 ng/ml vs. 55.46 ng/ml (range: 39.77 - 68.54 ng/ml in the mother's blood, which means that the level of TAFIa/ai antigen was significantly higher in fetal blood than in maternal blood (p<0.00001. TAFIa/ai antigen is an integral component of cord blood plasma. The concentration of TAFIa/ai antigen is about two times higher in fetal blood than in maternal blood.

  1. Development of a sandwich ELISA for the thrombin light chain identified by serum proteome analysis

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    Kazuyuki Sogawa

    2017-08-01

    Full Text Available We previously identified novel biomarker candidates in biliary tract cancer (BTC using serum proteome analysis. Among several candidates, we focused on thrombin light chain which is a 4204 Da peptide as the most promising biomarker for BTC. To move thrombin light chain toward potential diagnostic use, we developed an enzyme immunoassay that enables to measure serum thrombin light chain levels.Both one monoclonal antibody specific to the N-termini and one polyclonal antibody were used to develop a sandwich ELISA for thrombin light chain. The assay was evaluated by comparing the results with those obtained by the ClinProt™ system. Serum samples were obtained from 20 patients with BTC, 20 patients with BBTDs and 20 HVs using the ClinProt™ system and ELISA.The results of the established ELISA showed a positive correlation with the findings by ClinProt™ system (slope=0.3386, intercept=34.901, r2=0.9641. The performance of the ELISA was satisfactory in terms of recovery (97.9–102.5% and within-run (1.5–4.8% and between-day (1.9–6.7% reproducibility. Serum thrombin light chain levels were significantly greater in BTC (176.5±47.2 ng/mL than in BBTDs (128.6±17.4 ng/mL and HVs (127.6±16.0 ng/mL (p<0.001.The sandwich ELISA developed in this study will be useful for validation of the diagnostic significance of serum thrombin light chain levels in various cancers. Keywords: Thrombin light chain, Biliary tract cancer, Sandwich ELISA, Serum biomarker

  2. Percutaneous treatment of femoral pseudoaneurysms: comparison of fibrin sealant against thrombin

    Directory of Open Access Journals (Sweden)

    Daniel Mendes Pinto

    2013-12-01

    Full Text Available INTRODUCTION: Femoral pseudoaneurysms are a complication that occurs in connection with up to 8% of percutaneous procedures. Of the available treatments, ultrasound guided thrombin injection has a high success rate and is well-tolerated by patients. The combination of thrombin and fibrinogen known as fibrin sealant forms a stable clot and can be used to treat pseudoaneurysms, particularly those with complex anatomy and larger size. OBJECTIVE: To compare the results of treating femoral pseudoaneurysm in two ways: Group T was treated with thrombin alone and Group T+F was treated with fibrin sealant (thrombin+fibrinogen. METHODS: A retrospective analysis was conducted of femoral pseudoaneurysm cases treated between January 2005 and December 2012. RESULTS: Twenty-eight patients were treated, 21 with thrombin alone and seven with fibrin sealant. All patients in group T were treated successfully, but only four patients in group T+F were treated successfully (57.1% success rate in Group T+F, p<0.01. The three cases of failure in group T+F needed surgery and in one of these cases the complication was embolization to the femoral bifurcation. The pseudoaneurysms that were treated with fibrin sealant were larger (25 cm3 in Group T and 57.7 cm3 in Group T+F, p=0.02 and required larger volumes of thrombin (0.5 mL in Group T and 1.0 mL in Group T+F, p<0.01. There was one complication in Group T and two complications in Group T+F (p<0.01. CONCLUSIONS: Irrespective of the small number of cases reviewed, treatment with thrombin alone was superior to treating with fibrin sealant, since it caused few complications and was more effective at correcting pseudoaneurysms.

  3. Construction of photoelectrochemical thrombin aptasensor via assembling multilayer of graphene-CdS nanocomposites.

    Science.gov (United States)

    Shangguan, Li; Zhu, Wei; Xue, Yanchun; Liu, Songqin

    2015-02-15

    A photoelectrochemical (PEC) aptasensor for highly sensitive and specific detection of thrombin was developed by using graphene–CdS nanocomposites multilayer as photoactive species and electroactive mediator hexaammineruthenium(III) chloride (Ru(NH(3))(6)(3+)) as signal enhancer. Graphene–CdS nanocomposites (G–CdS) were synthesized by one-pot reduction of oxide graphene and CdCl2 with thioacetamide. The photoactive multilayer was prepared by alternative assembly of the negatively charged 3-mercaptopropionic acid modified graphene–CdS nanocomposites (MPA-G–CdS) and the positively charged polyethylenimine (PEI) on ITO electrode. This layer-by-layer assembly method enhanced the stability and homogeneity of the photocurrent readout of G–CdS. Thrombin aptamer was covalently bound to the multilayer by using glutaraldehyde as cross-linking. Electroactive mediator (Ru(NH(3))(6)(3+)) could interact with the DNA phosphate backbone and thus facilitated the electron transfer between G–CdS multilayer and electrode and enhanced the photocurrent. Hybridizing of a long complementary DNA with thrombin aptamer could increase the adsorption amount of (Ru(NH(3))(6)(3+)), which in turn boosted the signal readout. In the presence of target thrombin, the affinity interaction between thrombin and its aptamer resulted in the long complementary DNA releasing from the G–CdS multilayer and decreasing of photocurrent signal. On the basis of G–CdS multilayer as the photoactive species, (Ru (NH(3))(6)(3+)) as an electroactive mediator, and aptamer as a recognition module, a high sensitive PEC aptasensor for thrombin detection was proposed. The thrombin aptasensor displayed a linear range from 2.0 pM to 600.0 pM and a detection limit of 1.0 pM. The present strategy provided a promising ideology for the future development of PEC biosensor. Copyright © 2014 Elsevier B.V. All rights reserved.

  4. Design and characterization of hirulogs: A novel class of bivalent peptide inhibitors of thrombin

    Energy Technology Data Exchange (ETDEWEB)

    Maraganore, J.M.; Bourdon, P.; Jablonski, J.; Ramachandran, K.L. (Biogen, Inc., Cambridge, MA (USA)); Fenton, J.W. II (New York State Department of Health, Albany (USA))

    1990-07-31

    A novel class of synthetic peptides has been designed that inhibit the thrombin catalytic site and exhibit specificity for the anion-binding exosite (ABE) of {alpha}-thrombin. These peptides, called hirulogs, consist of (i) an active-site specificity sequence with a restricted Arg-Pro scissile bond, (ii) a polymeric linker of glycyl residues from 6 to 18 {angstrom} in length, and (iii) an ABE recognition sequence such as that in the hirudin C-terminus. Hirulog-1 ((D-Phe)-Pro-Arg-Pro-(Gly){sub 4}-Asn-Gly-Asp-Phe-Glu-Glu-Ile-Pro-Glu-Tyr-Leu) inhibits the thrombin-catalyzed hydrolysis of a tripeptide p-nitroanilide substrate with K{sub i} = 2.3 nM. In contrast, the synthetic C-terminal hirudin peptide S-Hir{sub 53-64}, which binds to the thrombin ABE, blocked the fibrinogen clotting activity of the enzyme with K{sub i} = 144 nM but failed to inhibit the hydrolysis of p-nitroanilide substrates at concentrations as high as 1 mM. Hirulog-1, but not S-Hir{sub 53-64}, was found to inhibit the incorporation of ({sup 14}C)diisopropyl fluorophosphate in thrombin. Hirulog-1 appears specific for thrombin as it lacks inhibitory activities toward human factor Xa, human plasmin, and bovine trypsin at inhibitor:enzyme concentrations 3 orders of magnitude higher than those required to inhibit thrombin. The optimal inhibitory activity of hirulog-1 depends upon all three components of its structure. Comparison of anticoagulant activities of hirulog-1, hirudin, and S-Hir{sub 53-64} showed that the synthetic hirulog-1 is 2-fold more potent than hirudin and 100-fold more active than S-Hir{sub 53-64} in increasing the activated partial thromboplastin time of normal human plasma.

  5. Robot-assisted automatic ultrasound calibration.

    Science.gov (United States)

    Aalamifar, Fereshteh; Cheng, Alexis; Kim, Younsu; Hu, Xiao; Zhang, Haichong K; Guo, Xiaoyu; Boctor, Emad M

    2016-10-01

    Ultrasound (US) calibration is the process of determining the unknown transformation from a coordinate frame such as the robot's tooltip to the US image frame and is a necessary task for any robotic or tracked US system. US calibration requires submillimeter-range accuracy for most applications, but it is a time-consuming and repetitive task. We provide a new framework for automatic US calibration with robot assistance and without the need for temporal calibration. US calibration based on active echo (AE) phantom was previously proposed, and its superiority over conventional cross-wire phantom-based calibration was shown. In this work, we use AE to guide the robotic arm motion through the process of data collection; we combine the capability of the AE point to localize itself in the frame of the US image with the automatic motion of the robotic arm to provide a framework for calibrating the arm to the US image automatically. We demonstrated the efficacy of the automated method compared to the manual method through experiments. To highlight the necessity of frequent ultrasound calibration, it is demonstrated that the calibration precision changed from 1.67 to 3.20 mm if the data collection is not repeated after a dismounting/mounting of the probe holder. In a large data set experiment, similar reconstruction precision of automatic and manual data collection was observed, while the time was reduced by 58 %. In addition, we compared ten automatic calibrations with ten manual ones, each performed in 15 min, and showed that all the automatic ones could converge in the case of setting the initial matrix as identity, while this was not achieved by manual data sets. Given the same initial matrix, the repeatability of the automatic was [0.46, 0.34, 0.80, 0.47] versus [0.42, 0.51, 0.98, 1.15] mm in the manual case for the US image four corners. The submillimeter accuracy requirement of US calibration makes frequent data collections unavoidable. We proposed an automated

  6. Random Forests Are Able to Identify Differences in Clotting Dynamics from Kinetic Models of Thrombin Generation.

    Science.gov (United States)

    Arumugam, Jayavel; Bukkapatnam, Satish T S; Narayanan, Krishna R; Srinivasa, Arun R

    2016-01-01

    Current methods for distinguishing acute coronary syndromes such as heart attack from stable coronary artery disease, based on the kinetics of thrombin formation, have been limited to evaluating sensitivity of well-established chemical species (e.g., thrombin) using simple quantifiers of their concentration profiles (e.g., maximum level of thrombin concentration, area under the thrombin concentration versus time curve). In order to get an improved classifier, we use a 34-protein factor clotting cascade model and convert the simulation data into a high-dimensional representation (about 19000 features) using a piecewise cubic polynomial fit. Then, we systematically find plausible assays to effectively gauge changes in acute coronary syndrome/coronary artery disease populations by introducing a statistical learning technique called Random Forests. We find that differences associated with acute coronary syndromes emerge in combinations of a handful of features. For instance, concentrations of 3 chemical species, namely, active alpha-thrombin, tissue factor-factor VIIa-factor Xa ternary complex, and intrinsic tenase complex with factor X, at specific time windows, could be used to classify acute coronary syndromes to an accuracy of about 87.2%. Such a combination could be used to efficiently assay the coagulation system.

  7. Thermodynamic, Anticoagulant, and Antiproliferative Properties of Thrombin Binding Aptamer Containing Novel UNA Derivative

    Directory of Open Access Journals (Sweden)

    Weronika Kotkowiak

    2018-03-01

    Full Text Available Thrombin is a serine protease that plays a crucial role in hemostasis, fibrinolysis, cell proliferation, and migration. Thrombin binding aptamer (TBA is able to inhibit the activity of thrombin molecule via binding to its exosite I. This 15-nt DNA oligonucleotide forms an intramolecular, antiparallel G-quadruplex structure with a chair-like conformation. In this paper, we report on our investigations on the influence of certain modified nucleotide residues on thermodynamic stability, folding topology, and biological properties of TBA variants. In particular, the effect of single incorporation of a novel 4-thiouracil derivative of unlocked nucleic acid (UNA, as well as single incorporation of 4-thiouridine and all four canonical UNAs, was evaluated. The studies presented herein have shown that 4-thiouridine in RNA and UNA series, as well as all four canonical UNAs, can efficiently modulate G-quadruplex thermodynamic and biological stability, and that the effect is strongly position dependent. Interestingly, TBA variants containing the modified nucleotide residues are characterized by unchanged folding topology. Thrombin time assay revealed that incorporation of certain UNA residues may improve G-quadruplex anticoagulant properties. Noteworthy, some TBA variants, characterized by decreased ability to inhibit thrombin activity, possess significant antiproliferative properties reducing the viability of the HeLa cell line even by 95% at 10 μM concentration.

  8. Progestin and thrombin regulate tissue factor expression in human term decidual cells.

    Science.gov (United States)

    Lockwood, C J; Murk, W; Kayisli, U A; Buchwalder, L F; Huang, S-T; Funai, E F; Krikun, G; Schatz, F

    2009-06-01

    Perivascular cell membrane-bound tissue factor (TF) initiates hemostasis via thrombin generation. The identity and potential regulation of TF-expressing cells at the human maternal-fetal interface that confers hemostatic protection during normal and preterm delivery is unclear. The objective of the study were to identify TF-expressing cells at the maternal-fetal interface in term and preterm decidual sections by immunohistochemistry and evaluate progestin, thrombin, TNF-alpha, and IL-1beta effects on TF expression by cultured human term decidual cells (DCs). Serial placental sections were immunostained for TF. Leukocyte-free term DC monolayers were incubated with 10(-8) M estradiol (E2) or E2 plus 10(-7) M medroxyprogestrone acetate (MPA) +/- thrombin or TNF-alpha or IL-1beta. ELISA and Western blotting assessed TF in cell lysates. Quantitative real-time RT-PCR measured TF mRNA levels. Immunolocalized TF in DC membranes in preterm and term placental sections displayed higher Histologic Scores than villous mesenchymal cells (P term placental sections, DC-expressed TF exceeds that of other cell types at the maternal-fetal interface and is localized at the cell membranes in which it can bind to factor VII and meet the hemostatic demands of labor and delivery via thrombin formation. Unlike the general concept that TF is constitutive in cells that highly express it, MPA and thrombin significantly enhanced TF expression in term DC monolayers.

  9. Random Forests Are Able to Identify Differences in Clotting Dynamics from Kinetic Models of Thrombin Generation.

    Directory of Open Access Journals (Sweden)

    Jayavel Arumugam

    Full Text Available Current methods for distinguishing acute coronary syndromes such as heart attack from stable coronary artery disease, based on the kinetics of thrombin formation, have been limited to evaluating sensitivity of well-established chemical species (e.g., thrombin using simple quantifiers of their concentration profiles (e.g., maximum level of thrombin concentration, area under the thrombin concentration versus time curve. In order to get an improved classifier, we use a 34-protein factor clotting cascade model and convert the simulation data into a high-dimensional representation (about 19000 features using a piecewise cubic polynomial fit. Then, we systematically find plausible assays to effectively gauge changes in acute coronary syndrome/coronary artery disease populations by introducing a statistical learning technique called Random Forests. We find that differences associated with acute coronary syndromes emerge in combinations of a handful of features. For instance, concentrations of 3 chemical species, namely, active alpha-thrombin, tissue factor-factor VIIa-factor Xa ternary complex, and intrinsic tenase complex with factor X, at specific time windows, could be used to classify acute coronary syndromes to an accuracy of about 87.2%. Such a combination could be used to efficiently assay the coagulation system.

  10. Marine Diterpenes: Molecular Modeling of Thrombin Inhibitors with Potential Biotechnological Application as an Antithrombotic

    Directory of Open Access Journals (Sweden)

    Rebeca Cristina Costa Pereira

    2017-03-01

    Full Text Available Thrombosis related diseases are among the main causes of death and incapacity in the world. Despite the existence of antithrombotic agents available for therapy, they still present adverse effects like hemorrhagic risks which justify the search for new options. Recently, pachydictyol A, isopachydictyol A, and dichotomanol, three diterpenes isolated from Brazilian marine brown alga Dictyota menstrualis were identified as potent antithrombotic molecules through inhibition of thrombin, a key enzyme of coagulation cascade and a platelet agonist. Due to the biotechnological potential of these marine metabolites, in this work we evaluated their binding mode to thrombin in silico and identified structural features related to the activity in order to characterize their molecular mechanism. According to our theoretical studies including structure-activity relationship and molecular docking analysis, the highest dipole moment, polar surface area, and lowest electronic density of dichotomanol are probably involved in its higher inhibition percentage towards thrombin catalytic activity compared to pachydictyol A and isopachydictyol A. Interestingly, the molecular docking studies also revealed a good shape complementarity of pachydictyol A and isopachydictyol A and interactions with important residues and regions (e.g., H57, S195, W215, G216, and loop-60, which probably justify their thrombin inhibitor effects demonstrated in vitro. Finally, this study explored the structural features and binding mode of these three diterpenes in thrombin which reinforced their potential to be further explored and may help in the design of new antithrombotic agents.

  11. Bufadienolides from Kalanchoe daigremontiana as thrombin inhibitors-In vitro and in silico study.

    Science.gov (United States)

    Kolodziejczyk-Czepas, Joanna; Sieradzka, Malgorzata; Moniuszko-Szajwaj, Barbara; Pecio, Łukasz; Ponczek, Michal B; Nowak, Pawel; Stochmal, Anna

    2017-06-01

    Thrombin is an active plasma coagulation factor II, critical for the formation of fibrin clot during blood coagulation. For that reason, this protein is also a crucial target for different anti-thrombotic therapies. The work is based on in vitro evaluation of the inhibitory effect of bufadienolide-rich fraction, isolated from roots of Kalanchoe daigremontiana (1-50μg/ml) on enzymatic properties of a serine proteinase - thrombin. The efficacy of the inhibition of amidolytic activity of thrombin (measured as a hydrolysis of the chromogenic substrate S-2238, Chromogenix) attained about 10 and 66%, respectively. The IC 50 , established for the examined bufadienolide fraction was 2.79μg/ml, while the IC 50 calculated for argatroban (reference compound) was 0.78μg/ml. Linearization conducted using Lineweaver-Burk plot indicated that the K. daigremontiana fraction contains compounds that are uncompetitive inhibitors of thrombin. K. daigremontiana fraction was also able to reduce the proteolytic activity of thrombin towards its physiological substrate, i.e. fibrinogen. Additionally, this study is supported by in silico analysis of interactions of the most common compounds, identified in the examined in Kalanchoe extract to crystal structure of this enzyme. Copyright © 2017 Elsevier B.V. All rights reserved.

  12. Platelet-derived microparticles regulates thrombin generation via phophatidylserine in abdominal sepsis.

    Science.gov (United States)

    Wang, Yongzhi; Zhang, Su; Luo, Lingtao; Norström, Eva; Braun, Oscar Ö; Mörgelin, Matthias; Thorlacius, Henrik

    2018-02-01

    Sepsis is associated with dysfunctional coagulation. Recent data suggest that platelets play a role in sepsis by promoting neutrophil accumulation. Herein, we show that cecal ligation and puncture (CLP) triggered systemic inflammation, which is characterized by formation of IL-6 and CXC chemokines as well as neutrophil accumulation in the lung. Platelet depletion decreased neutrophil accumulation, IL-6, and CXC chemokines formation in septic lungs. Depletion of platelets increased peak thrombin formation and total thrombin generation (TG) in plasma from septic animals. CLP elevated circulating levels of platelet-derived microparticles (PMPs). In vitro generated PMPs were a potent inducer of TG. Interestingly, in vitro wild-type recombinant annexin V abolished PMP-induced thrombin formation whereas a mutant annexin V protein, which does not bind to phosphatidylserine (PS), had no effect. Administration of wild-type, but not mutant annexin V, significantly inhibited thrombin formation in septic animals. Moreover, CLP-induced formation of thrombin-antithrombin complexes were reduced in platelet-depleted mice and in animals pretreated with annexin V. PMP-induced TG attenuated in FXII- and FVII-deficient plasma. These findings suggest that sepsis-induced TG is dependent on platelets. Moreover, PMPs formed in sepsis are a potent inducer of TG via PS exposure, and activation of both the intrinsic and extrinsic pathway of coagulation. In conclusion, these observations suggest that PMPs and PS play an important role in dysfunctional coagulation in abdominal sepsis. © 2017 Wiley Periodicals, Inc.

  13. Genetic Determinants of Thrombin Generation and Their Relation to Venous Thrombosis: Results from the GAIT-2 Project

    Science.gov (United States)

    Martin-Fernandez, Laura; Ziyatdinov, Andrey; Carrasco, Marina; Millon, Juan Antonio; Martinez-Perez, Angel; Vilalta, Noelia; Brunel, Helena; Font, Montserrat; Hamsten, Anders; Souto, Juan Carlos; Soria, José Manuel

    2016-01-01

    Background Venous thromboembolism (VTE) is a common disease where known genetic risk factors explain only a small portion of the genetic variance. Then, the analysis of intermediate phenotypes, such as thrombin generation assay, can be used to identify novel genetic risk factors that contribute to VTE. Objectives To investigate the genetic basis of distinct quantitative phenotypes of thrombin generation and its relationship to the risk of VTE. Patients/Methods Lag time, thrombin peak and endogenous thrombin potential (ETP) were measured in the families of the Genetic Analysis of Idiopathic Thrombophilia 2 (GAIT-2) Project. This sample consisted of 935 individuals in 35 extended families selected through a proband with idiopathic thrombophilia. We performed also genome wide association studies (GWAS) with thrombin generation phenotypes. Results The results showed that 67% of the variation in the risk of VTE is attributable to genetic factors. The heritabilities of lag time, thrombin peak and ETP were 49%, 54% and 52%, respectively. More importantly, we demonstrated also the existence of positive genetic correlations between thrombin peak or ETP and the risk of VTE. Moreover, the major genetic determinant of thrombin generation was the F2 gene. However, other suggestive signals were observed. Conclusions The thrombin generation phenotypes are strongly genetically determined. The thrombin peak and ETP are significantly genetically correlated with the risk of VTE. In addition, F2 was identified as a major determinant of thrombin generation. We reported suggestive signals that might increase our knowledge to explain the variability of this important phenotype. Validation and functional studies are required to confirm GWAS results. PMID:26784699

  14. Substitution of valine for glycine-558 in the congenital dysthrombin thrombin Quick II alters primary substrate specificity

    Energy Technology Data Exchange (ETDEWEB)

    Henriksen, R.A.; Mann, K.G. (Univ. of Vermont, Burlington (USA))

    1989-03-07

    Thrombin Quick II is one of two dysfunctional forms of thrombin derived from the previously described congenital dysprothrombin prothrombin Quick. Thrombin Quick II does not clot fibrinogen, hydrolyze p-nitroanilide substrates of thrombin, or bind N{sup 2}-(5-(dimethylamino)naphthalene-1-sulfonyl)arginine N,N-(3-ethyl-1,5-pentanediyl)amide, a high-affinity competitive inhibitor of thrombin. To determine the structural alteration in thrombin Quick II, the reduced, carboxymethylated protein was hydrolyzed by a lysyl endopeptidase. A peptide not present in a parallel thrombin hydrolysate was identified by reverse-phase chromatography. This Gly residue, which is highly conserved in the chymotrypsin family of serine proteases, forms part of the substrate binding pocket for bulky aromatic and basic side chains in chymotrypsin and trypsin, respectively. However, in porcine elastase 1, the corresponding residue is threonine. Consistent with the identified structural alteration, thrombin Quick II incorporates ({sup 3}H)diisopropyl fluorophosphate stoichiometrically and hydrolyzes the elastase substrate succinyl-Ala-Ala-Pro-Leu-p-nitroanilide with a relative k{sub cat}/K{sub M} of 0.14 when compared to thrombin. This results from a 3-fold increase in K{sub M} and a 2.5-fold decrease in k{sub cat} for thrombin Quick II when compared to thrombin acting on the same substrate. These results and those of other investigators studying mutant trypsins support the conclusion that the catalytic activity of serine proteases is very sensitive to structural alterations in the primary substrate binding pocket.

  15. Home Automation

    OpenAIRE

    Ahmed, Zeeshan

    2010-01-01

    In this paper I briefly discuss the importance of home automation system. Going in to the details I briefly present a real time designed and implemented software and hardware oriented house automation research project, capable of automating house's electricity and providing a security system to detect the presence of unexpected behavior.

  16. Three-point bridge calibration with one resistor

    Science.gov (United States)

    Harrison, D. R.; Brown, R. M.

    1974-01-01

    Method calibrates transducer bridge curing unbalanced condition and line resistance errors are negligible. Series resistance method can be automated easily and controlled by 2-bit information source which provide 4 states for switches.

  17. Automatic multi-camera calibration for deployable positioning systems

    Science.gov (United States)

    Axelsson, Maria; Karlsson, Mikael; Rudner, Staffan

    2012-06-01

    Surveillance with automated positioning and tracking of subjects and vehicles in 3D is desired in many defence and security applications. Camera systems with stereo or multiple cameras are often used for 3D positioning. In such systems, accurate camera calibration is needed to obtain a reliable 3D position estimate. There is also a need for automated camera calibration to facilitate fast deployment of semi-mobile multi-camera 3D positioning systems. In this paper we investigate a method for automatic calibration of the extrinsic camera parameters (relative camera pose and orientation) of a multi-camera positioning system. It is based on estimation of the essential matrix between each camera pair using the 5-point method for intrinsically calibrated cameras. The method is compared to a manual calibration method using real HD video data from a field trial with a multicamera positioning system. The method is also evaluated on simulated data from a stereo camera model. The results show that the reprojection error of the automated camera calibration method is close to or smaller than the error for the manual calibration method and that the automated calibration method can replace the manual calibration.

  18. Use of a robotic manipulator in the simulation of the automation of a calibration process of dosemeters; Uso de un manipulador robotico en la simulacion de la automatizacion de un proceso de calibracion de dosimetros

    Energy Technology Data Exchange (ETDEWEB)

    Benitez R, J S; Najera H, M C [Instituto Nacional de Investigaciones Nucleares, A.P. 18-1027, 11801 Mexico D.F. (Mexico)

    2002-07-01

    The development of a system based in a manipulative robot which simulates the operative sequence in a calibration process of dosemeters is presented. In this process it is performed the monitoring of the dosemeter positions and the calibrator by mean of an arm of articulated robot which develops the movement sequences and the taking a decision based on the information coming from the external sensors. (Author)

  19. Modeling the microscopic electrical properties of thrombin binding aptamer (TBA) for label-free biosensors

    Science.gov (United States)

    Alfinito, Eleonora; Reggiani, Lino; Cataldo, Rosella; De Nunzio, Giorgio; Giotta, Livia; Guascito, Maria Rachele

    2017-02-01

    Aptamers are chemically produced oligonucleotides, able to bind a variety of targets such as drugs, proteins and pathogens with high sensitivity and selectivity. Therefore, aptamers are largely employed for producing label-free biosensors (aptasensors), with significant applications in diagnostics and drug delivery. In particular, the anti-thrombin aptamers are biomolecules of high interest for clinical use, because of their ability to recognize and bind the thrombin enzyme. Among them, the DNA 15-mer aptamer (TBA), has been widely explored around the possibility of using it in aptasensors. This paper proposes a microscopic model of the electrical properties of TBA and of the aptamer-thrombin complex, combining information from both structure and function, following the issues addressed in an emerging branch of electronics known as proteotronics. The theoretical results are compared and validated with measurements reported in the literature. Finally, the model suggests resistance measurements as a novel tool for testing aptamer-target affinity.

  20. Binding of poly(amidoamine), carbosilane, phosphorus and hybrid dendrimers to thrombin-Constants and mechanisms.

    Science.gov (United States)

    Shcharbin, Dzmitry; Pedziwiatr-Werbicka, Elzbieta; Vcherashniaya, Aliaksandra; Janaszewska, Anna; Marcinkowska, Monika; Goska, Piotr; Klajnert-Maculewicz, Barbara; Ionov, Maksim; Abashkin, Viktar; Ihnatsyeu-Kachan, Aliaksei; de la Mata, F Javier; Ortega, Paula; Gomez-Ramirez, Rafael; Majoral, Jean-Pierre; Bryszewska, Maria

    2017-07-01

    Thrombin is an essential part of the blood coagulation system; it is a serine protease that converts soluble fibrinogen into insoluble strands of fibrin, and catalyzes many other coagulation-related reactions. Absorption at its surface of small nanoparticles can completely change the biological properties of thrombin. We have analyzed the influence on thrombin of 3 different kinds of small nanoparticles: dendrimers (phosphorus-based, carbosilane based and polyamidoamine) and 2 hybrid systems containing carbosilane, viologen and phosphorus dendritic scaffolds in one single molecule, bearing different flexibility, size and surface charge. There was significant alteration in the rigidity of the rigid dendrimers in contrast to flexible dendrimers. These differences in their action are important in understanding interactions taking place at a bio-nanointerface. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Placental vascular pathology and increased thrombin generation as mechanisms of disease in obstetrical syndromes

    Directory of Open Access Journals (Sweden)

    Salvatore Andrea Mastrolia

    2014-11-01

    Full Text Available Obstetrical complications including preeclampsia, fetal growth restriction, preterm labor, preterm prelabor rupture of membranes and fetal demise are all the clinical endpoint of several underlying mechanisms (i.e., infection, inflammation, thrombosis, endocrine disorder, immunologic rejection, genetic, and environmental, therefore, they may be regarded as syndromes. Placental vascular pathology and increased thrombin generation were reported in all of these obstetrical syndromes. Moreover, elevated concentrations of thrombin-anti thrombin III complexes and changes in the coagulation as well as anticoagulation factors can be detected in the maternal circulation prior to the clinical development of the disease in some of these syndromes. In this review, we will assess the changes in the hemostatic system during normal and complicated pregnancy in maternal blood, maternal–fetal interface and amniotic fluid, and describe the contribution of thrombosis and vascular pathology to the development of the great obstetrical syndromes.

  2. The effect of cigarette smoke extract on thrombomodulin-thrombin binding: an atomic force microscopy study.

    Science.gov (United States)

    Wei, Yujie; Zhang, Xuejie; Xu, Li; Yi, Shaoqiong; Li, Yi; Fang, Xiaohong; Liu, Huiliang

    2012-10-01

    Cigarette smoking is a well-known risk factor for cardiovascular disease. Smoking can cause vascular endothelial dysfunction and consequently trigger haemostatic activation and thrombosis. However, the mechanism of how smoking promotes thrombosis is not fully understood. Thrombosis is associated with the imbalance of the coagulant system due to endothelial dysfunction. As a vital anticoagulation cofactor, thrombomodulin (TM) located on the endothelial cell surface is able to regulate intravascular coagulation by binding to thrombin, and the binding results in thrombosis inhibition. This work focused on the effects of cigarette smoke extract (CSE) on TM-thrombin binding by atomic force microscopy (AFM) based single-molecule force spectroscopy. The results from both in vitro and live-cell experiments indicated that CSE could notably reduce the binding probability of TM and thrombin. This study provided a new approach and new evidence for studying the mechanism of thrombosis triggered by cigarette smoking.

  3. Thrombin immobilization to methacrylic acid grafted poly(3-hydroxybutyrate) and its in vitro application.

    Science.gov (United States)

    Akkaya, Alper; Pazarlioglu, Nurdan

    2013-01-01

    Poly(3-hydroxybutyrate) is nontoxic and biodegradable, with good biocompatibility and potential support for long-term implants. For this reason, it is a good support for enzyme immobilization. Enzyme immobilization could not be done directly because poly(3-hydroxybutyrate) has no functional groups. Therefore, modification should be done for enzyme immobilization. In this study, methacrylic acid was graft polymerized to poly(3-hydroxybutyrate) and thrombin was immobilized to polymethacrylic acid grafted poly(3-hydroxybutyrate). In fact, graft polymerization of methacrylic acid to poly(3-hydroxybutyrate) and thrombin immobilization was a model study. Biomolecule immobilized poly(3-hydroxybutyrate) could be used as an implant. Thrombin was selected as a biomolecule for this model study and it was immobilized to methacrylic acid grafted poly(3-hydroxybutyrate). Then the developed product was used to stop bleeding.

  4. Improved thrombin binding aptamer by incorporation of a single unlocked nucleic acid monomer

    DEFF Research Database (Denmark)

    Pasternak, Anna; Hernandez, Frank J; Rasmussen, Lars Melholt

    2011-01-01

    A 15-mer DNA aptamer (named TBA) adopts a G-quadruplex structure that strongly inhibits fibrin-clot formation by binding to thrombin. We have performed thermodynamic analysis, binding affinity and biological activity studies of TBA variants modified by unlocked nucleic acid (UNA) monomers. UNA...... that a UNA monomer is allowed in many positions of the aptamer without significantly changing the thrombin-binding properties. The biological effect of a selection of the modified aptamers was tested by a thrombin time assay and showed that most of the UNA-modified TBAs possess anticoagulant properties......, and that the construct with a UNA-U monomer in position 7 is a highly potent inhibitor of fibrin-clot formation....

  5. Pseudomonas aeruginosa elastase cleaves a C-terminal peptide from human thrombin that inhibits host inflammatory responses

    DEFF Research Database (Denmark)

    van der Plas, Mariena J A; Bhongir, Ravi K V; Kjellström, Sven

    2016-01-01

    Pseudomonas aeruginosa is an opportunistic pathogen known for its immune evasive abilities amongst others by degradation of a large variety of host proteins. Here we show that digestion of thrombin by P. aeruginosa elastase leads to the release of the C-terminal thrombin-derived peptide FYT21...

  6. Factor XI dependent and independent activation of thrombin activatable fibrinolysis inhibitor (TAFI) in plasma associated with clot formation

    NARCIS (Netherlands)

    Bouma, B. N.; Mosnier, L. O.; Meijers, J. C.; Griffin, J. H.

    1999-01-01

    Thrombin Activatable Fibrinolysis Inhibitor (TAFI) also known as plasma procarboxypeptidase B is activated by relatively high concentrations of thrombin in a reaction stimulated by thrombomodulin. In plasma an intact factor XI-dependent feed back loop via the intrinsic pathway is necessary to

  7. Thrombin impairs human endometrial endothelial angiogenesis; implications for progestin-only contraceptive-induced abnormal uterine bleeding.

    Science.gov (United States)

    Shapiro, John P; Guzeloglu-Kayisli, Ozlem; Kayisli, Umit A; Semerci, Nihan; Huang, S Joseph; Arlier, Sefa; Larsen, Kellie; Fadda, Paolo; Schatz, Frederick; Lockwood, Charles J

    2017-06-01

    Progestin-only contraceptives induce abnormal uterine bleeding, accompanied by prothrombin leakage from dilated endometrial microvessels and increased thrombin generation by human endometrial stromal cell (HESC)-expressed tissue factor. Initial studies of the thrombin-treated HESC secretome identified elevated levels of cleaved chondroitin sulfate proteoglycan 4 (CSPG4), impairing pericyte-endothelial interactions. Thus, we investigated direct and CSPG4-mediated effects of thrombin in eliciting abnormal uterine bleeding by disrupting endometrial angiogenesis. Liquid chromatography/tandem mass spectrometry, enzyme-linked immunosorbent assay (ELISA) and quantitative real-time-polymerase chain reaction (PCR) evaluated conditioned medium supernatant and cell lysates from control versus thrombin-treated HESCs. Pre- and post-Depo medroxyprogesterone acetate (DMPA)-administered endometria were immunostained for CSPG4. Proliferation, apoptosis and tube formation were assessed in human endometrial endothelial cells (HEECs) incubated with recombinant human (rh)-CSPG4 or thrombin or both. Thrombin induced CSPG4 protein expression in cultured HESCs as detected by mass spectrometry and ELISA (pabnormal uterine bleeding in DMPA users. Mass spectrometry analysis identified several HESC-secreted proteins regulated by thrombin. Therapeutic agents blocking angiogenic effects of thrombin in HESCs can prevent or minimize progestin-only contraceptive-induced abnormal uterine bleeding. Copyright © 2017. Published by Elsevier Inc.

  8. Aptamer-based turn-on fluorescent four-branched quaternary ammonium pyrazine probe for selective thrombin detection.

    Science.gov (United States)

    Yan, Shengyong; Huang, Rong; Zhou, Yangyang; Zhang, Ming; Deng, Minggang; Wang, Xiaolin; Weng, Xiaocheng; Zhou, Xiang

    2011-01-28

    In this thrombin detection system, the bright fluorescence of TASPI is almost eliminated by the DNA aptamer TBA (turn-off); however, in the presence of thrombin, it specifically binds to TBA by folding unrestricted TBA into an anti-parallel G-quadruplex structure and then releasing TASPI molecules, resulting in vivid and facile fluorescence recovery (turn-on).

  9. Systems biology of coagulation initiation: kinetics of thrombin generation in resting and activated human blood.

    Directory of Open Access Journals (Sweden)

    Manash S Chatterjee

    2010-09-01

    Full Text Available Blood function defines bleeding and clotting risks and dictates approaches for clinical intervention. Independent of adding exogenous tissue factor (TF, human blood treated in vitro with corn trypsin inhibitor (CTI, to block Factor XIIa will generate thrombin after an initiation time (T(i of 1 to 2 hours (depending on donor, while activation of platelets with the GPVI-activator convulxin reduces T(i to ∼20 minutes. Since current kinetic models fail to generate thrombin in the absence of added TF, we implemented a Platelet-Plasma ODE model accounting for: the Hockin-Mann protease reaction network, thrombin-dependent display of platelet phosphatidylserine, VIIa function on activated platelets, XIIa and XIa generation and function, competitive thrombin substrates (fluorogenic detector and fibrinogen, and thrombin consumption during fibrin polymerization. The kinetic model consisting of 76 ordinary differential equations (76 species, 57 reactions, 105 kinetic parameters predicted the clotting of resting and convulxin-activated human blood as well as predicted T(i of human blood under 50 different initial conditions that titrated increasing levels of TF, Xa, Va, XIa, IXa, and VIIa. Experiments with combined anti-XI and anti-XII antibodies prevented thrombin production, demonstrating that a leak of XIIa past saturating amounts of CTI (and not "blood-borne TF" alone was responsible for in vitro initiation without added TF. Clotting was not blocked by antibodies used individually against TF, VII/VIIa, P-selectin, GPIb, protein disulfide isomerase, cathepsin G, nor blocked by the ribosome inhibitor puromycin, the Clk1 kinase inhibitor Tg003, or inhibited VIIa (VIIai. This is the first model to predict the observed behavior of CTI-treated human blood, either resting or stimulated with platelet activators. CTI-treated human blood will clot in vitro due to the combined activity of XIIa and XIa, a process enhanced by platelet activators and which proceeds

  10. Thrombin like activity of Asclepias curassavica L. latex: action of cysteine proteases.

    Science.gov (United States)

    Shivaprasad, H V; Rajesh, R; Nanda, B L; Dharmappa, K K; Vishwanath, B S

    2009-05-04

    To validate the scientific basis of plant latex to stop bleeding on fresh cuts. Cysteine protease(s) from Asclepias curassavica (Asclepiadaceae) plant latex was assessed for pro-coagulant and thrombin like activities. A waxy material from the latex of Asclepias curassavica latex was removed by freezing and thawing. The resulted latex enzyme fraction was assayed for proteolytic activity using denatured casein as substrate. Its coagulant activity and thrombin like activity were determined using citrated plasma and pure fibrinogen, respectively. Inhibition studies were performed using specific protease inhibitors to know the type of protease. The latex enzyme fraction exhibited strong proteolytic activity when compared to trypsin and exerted pro-coagulant action by reducing plasma clotting time from 195 to 58 s whereas trypsin reduced clotting time marginally from 195 to 155 s. The pro-coagulant activity of this enzyme fraction was exerted by selectively hydrolyzing A alpha and B beta subunits of fibrinogen to form fibrin clot when pure fibrinogen was used as substrate as assessed by fibrinogen-agarose plate method and fibrinogen polymerization assay. Trypsin failed to induce any fibrin clot under similar conditions. The electrophoretic pattern of latex enzyme fraction-induced fibrin clot was very much similar to that of thrombin-induced fibrin clot and mimic thrombin like action. The proteolytic activity including thrombin like activity of Asclepias curassavica latex enzyme fraction was completely inhibited by iodoaceticacid (IAA). Cysteine proteases from Asclepias curassavica latex exhibited strong pro-coagulant action and were found to be specific in its action (Thrombin like). This could be the basis for the use of plant latex in pharmacological applications that justify their use as folk medicine.

  11. Percutaneous Thrombin Injection to Complete SMA Pseudoaneurysm Exclusion After Failing of Endograft Placement

    International Nuclear Information System (INIS)

    Szopinski, Piotr; Ciostek, Piotr; Pleban, Eliza; Iwanowski, Jaroslaw; Krol, Malgorzata Serafin-; Marianowska, Agnieszka; Noszczyk, Wojciech

    2005-01-01

    Visceral aneurysms are potentially life-threatening vascular lesions. Superior mesenteric artery (SMA) pseudoaneurysms are a rare but well-recognized complication of chronic pancreatitis. Open surgical repair of such an aneurysm, especially in patients after previous surgical treatment, might be dangerous and risky. Stent graft implantation makes SMA pseudoaneurysm exclusion possible and therefore avoids a major abdominal operation. Percutaneous direct thrombin injection is also one of the methods of treating aneurysms in this area. We report a first case of percutaneous ultrasound-guided thrombin injection to complete SMA pseudoaneurysm exclusion after an unsuccessful endograft placement. Six-month follow-up did not demonstrate any signs of aneurysm recurrence

  12. Structural Characterization of a Thrombin-Aptamer Complex by High Resolution Native Top-Down Mass Spectrometry

    Science.gov (United States)

    Zhang, Jiang; Loo, Rachel R. Ogorzalek; Loo, Joseph A.

    2017-09-01

    Native mass spectrometry (MS) with electrospray ionization (ESI) has evolved as an invaluable tool for the characterization of intact native proteins and non-covalently bound protein complexes. Here we report the structural characterization by high resolution native top-down MS of human thrombin and its complex with the Bock thrombin binding aptamer (TBA), a 15-nucleotide DNA with high specificity and affinity for thrombin. Accurate mass measurements revealed that the predominant form of native human α-thrombin contains a glycosylation mass of 2205 Da, corresponding to a sialylated symmetric biantennary oligosaccharide structure without fucosylation. Native MS showed that thrombin and TBA predominantly form a 1:1 complex under near physiological conditions (pH 6.8, 200 mM NH4OAc), but the binding stoichiometry is influenced by the solution ionic strength. In 20 mM ammonium acetate solution, up to two TBAs were bound to thrombin, whereas increasing the solution ionic strength destabilized the thrombin-TBA complex and 1 M NH4OAc nearly completely dissociated the complex. This observation is consistent with the mediation of thrombin-aptamer binding through electrostatic interactions and it is further consistent with the human thrombin structure that contains two anion binding sites on the surface. Electron capture dissociation (ECD) top-down MS of the thrombin-TBA complex performed with a high resolution 15 Tesla Fourier transform ion cyclotron resonance (FTICR) mass spectrometer showed the primary binding site to be at exosite I located near the N-terminal sequence of the heavy chain, consistent with crystallographic data. High resolution native top-down MS is complementary to traditional structural biology methods for structurally characterizing native proteins and protein-DNA complexes. [Figure not available: see fulltext.

  13. Longitudinal assessment of thrombin generation potential in response to alteration of antiplatelet therapy after TIA or ischaemic stroke.

    LENUS (Irish Health Repository)

    Tobin, W O

    2013-02-01

    The impact of changing antiplatelet therapy on thrombin generation potential in patients with ischaemic cerebrovascular disease (CVD) is unclear. We assessed patients within 4 weeks of TIA or ischaemic stroke (baseline), and then 14 days (14d) and >90 days (90d) after altering antiplatelet therapy. Thrombin generation was assessed in platelet poor plasma. Ninety-one patients were recruited. Twenty-four were initially assessed on no antiplatelet therapy, and then after 14d (N = 23) and 90d (N = 8) on aspirin monotherapy; 52 were assessed on aspirin monotherapy, and after 14 and 90 days on aspirin and dipyridamole combination therapy; 21 patients were assessed on aspirin and after 14 days (N = 21) and 90 days (N = 19) on clopidogrel. Peak thrombin generation and endogenous thrombin potential were reduced at 14 and 90 days (p ≤ 0.04) in the overall cohort. We assessed the impact of individual antiplatelet regimens on thrombin generation parameters to investigate the cause of this effect. Lag time and time-to-peak thrombin generation were unchanged at 14 days, but reduced 90 days after commencing aspirin (p ≤ 0.009). Lag time, peak thrombin generation and endogenous thrombin potential were reduced at both 14 and 90 days after adding dipyridamole to aspirin (p ≤ 0.01). Lag time was reduced 14 days after changing from aspirin to clopidogrel (p = 0.045), but this effect was not maintained at 90 days (p = 0.2). This pilot study did not show any consistent effects of commencing aspirin, or of changing from aspirin to clopidogrel on thrombin generation potential during follow-up. The addition of dipyridamole to aspirin led to a persistent reduction in peak and total thrombin generation ex vivo, and illustrates the diverse, potentially beneficial, newly recognised \\'anti-coagulant\\' effects of dipyridamole in ischaemic CVD.

  14. Geometric calibration of ERS satellite SAR images

    DEFF Research Database (Denmark)

    Mohr, Johan Jacob; Madsen, Søren Nørvang

    2001-01-01

    Geometric calibration of the European Remote Sensing (ERS) Satellite synthetic aperture radar (SAR) slant range images is important in relation to mapping areas without ground reference points and also in relation to automated processing. The relevant SAR system parameters are discussed...

  15. Development and implementation of an automated system for antiquated of the process of gamma radiation monitors calibration; Desenvolvimento e implantacao de um sistema automatizado para adequacao do processo de calibracao de monitores de radiacao gama

    Energy Technology Data Exchange (ETDEWEB)

    Silva Junior, Iremar Alves

    2012-07-01

    In this study it was carried out the development and implementation of a system for the appropriate process of gamma radiation monitors calibration, constituted by a pneumatic dispositive to exchange the attenuators and a positioning table, both actuated through a control panel. We also implemented a System of Caesa-Gammatron Irradiator, which increased the range of the air kerma rates, due to its higher activity comparing with the current system of gamma radiation in use in the calibration laboratory of gamma irradiation. Hence, it was necessary the installation of an attenuator dispositive remotely controlled in this irradiator system. Lastly, it was carried out an evaluation of the reduction in the rates of the occupational dose. This dissertation was developed with the aim of improving the quality of the services of calibration and tests of gamma radiation monitors - provided by the IPEN Laboratory of Instrument Calibration - as well as decreasing the occupational dose of the technicians involved in the process of calibration, following thus the principles of radiation protection. (author)

  16. Rigidification of the autolysis loop enhances Na[superscript +] binding to thrombin

    Energy Technology Data Exchange (ETDEWEB)

    Pozzi, Nicola; Chen, Raymond; Chen, Zhiwei; Bah, Alaji; Di Cera, Enrico (St. Louis-MED)

    2011-09-20

    Binding of Na{sup +} to thrombin ensures high activity toward physiological substrates and optimizes the procoagulant and prothrombotic roles of the enzyme in vivo. Under physiological conditions of pH and temperature, the binding affinity of Na{sup +} is weak due to large heat capacity and enthalpy changes associated with binding, and the K{sub d} = 80 mM ensures only 64% saturation of the site at the concentration of Na{sup +} in the blood (140 mM). Residues controlling Na{sup +} binding and activation have been identified. Yet, attempts to improve the interaction of Na{sup +} with thrombin and possibly increase catalytic activity under physiological conditions have so far been unsuccessful. Here we report how replacement of the flexible autolysis loop of human thrombin with the homologous rigid domain of the murine enzyme results in a drastic (up to 10-fold) increase in Na{sup +} affinity and a significant improvement in the catalytic activity of the enzyme. Rigidification of the autolysis loop abolishes the heat capacity change associated with Na{sup +} binding observed in the wild-type and also increases the stability of thrombin. These findings have general relevance to protein engineering studies of clotting proteases and trypsin-like enzymes.

  17. Label-free aptamer biosensor for thrombin detection based on functionalized graphene nanocomposites.

    Science.gov (United States)

    Wang, Qingqing; Zhou, Zhixue; Zhai, Yanling; Zhang, Lingling; Hong, Wei; Zhang, Zhiquan; Dong, Shaojun

    2015-08-15

    A label-free and amplified electrochemical impedimetric aptasensor based on functionalized graphene nanocomposites (rGO-AuNPs) was developed for the detection of thrombin, which played a vital role in thrombosis and hemostasis. The thiolated aptamer and dithiothreitol (TBA15-DTT) were firstly immobilized on the gold electrode to capture the thrombin molecules, and then aptamer functionalized graphene nanocomposites (rGO-TBA29) were used to fabricate a sandwich sensing platform for amplifying the impedimetric signals. As numerous negative charges of TBA29 on the electrode repelled to the [Fe(CN)6](4-/3-) anions, resulting in an obvious amplified charge-transfer resistance (Rct) signal. The Rct increase was linearly proportional to the thrombin concentration from 0.3 to 50nM and a detection limit of 0.01nM thrombin was achieved. In addition, graphene could also be labeled with other probes via electrostatic or π-π stacking interactions to produce signals, therefore different detection methods expanding wide application could be used in this model. Copyright © 2015. Published by Elsevier B.V.

  18. Regulation of Thrombin-Induced Lung Endothelial Cell Barrier Disruption by Protein Kinase C Delta.

    Directory of Open Access Journals (Sweden)

    Lishi Xie

    Full Text Available Protein Kinase C (PKC plays a significant role in thrombin-induced loss of endothelial cell (EC barrier integrity; however, the existence of more than 10 isozymes of PKC and tissue-specific isoform expression has limited our understanding of this important second messenger in vascular homeostasis. In this study, we show that PKCδ isoform promotes thrombin-induced loss of human pulmonary artery EC barrier integrity, findings substantiated by PKCδ inhibitory studies (rottlerin, dominant negative PKCδ construct and PKCδ silencing (siRNA. In addition, we identified PKCδ as a signaling mediator upstream of both thrombin-induced MLC phosphorylation and Rho GTPase activation affecting stress fiber formation, cell contraction and loss of EC barrier integrity. Our inhibitor-based studies indicate that thrombin-induced PKCδ activation exerts a positive feedback on Rho GTPase activation and contributes to Rac1 GTPase inhibition. Moreover, PKD (or PKCμ and CPI-17, two known PKCδ targets, were found to be activated by PKCδ in EC and served as modulators of cytoskeleton rearrangement. These studies clarify the role of PKCδ in EC cytoskeleton regulation, and highlight PKCδ as a therapeutic target in inflammatory lung disorders, characterized by the loss of barrier integrity, such as acute lung injury and sepsis.

  19. Percutaneous thrombin injection for the treatment of a post-pancreatitis pseudoaneurysm

    International Nuclear Information System (INIS)

    Puri, S.; Nicholson, A.A.; Breen, D.J.

    2003-01-01

    Visceral artery pseudoaneurysms are often treated surgically or by transcatheter embolisation. We report a case of a pseudoaneurysm in a patient with chronic pancreatitis, which was successfully occluded by percutaneous injection of thrombin into the pseudoaneurysmal sac as a first-line management. (orig.)

  20. EFFICACY OF THROMBIN FIBRIN GLUE AND SCLE ROSANT IN THE MANAGEMENT OF BLEEDI NG GASTRIC VARICES

    Directory of Open Access Journals (Sweden)

    Sanjay Gupta

    2015-01-01

    Full Text Available Gastric varices are noted in up to 20 % of patents with portal hypertension , and are more common in those with non - cirrhotic etiology 1 . They bleed at lower portal pressures , bleed more severely and are associated with higher rates of rebleed , encephalopathy and mortality 1,2,3 . Variceal obliteration using tissue adhesives such as N - butyl cyanoacrylate leading to plugging and thrombosis of the gastric varices is currently the first line management option for obliteration of the gastric varices 3 . Although various options have been proposed , gold standard for management of gastric variceal bleeds is yet to be defined. We theorized that injection of the gastric varices using thrombin based glue followed by injection of a sclerosant shall be effective in optimum sclerotherapy and eradication of gastric varices. MATERIAL AND METHODS : All patients presenting with gastric variceal bleed were offered sclerotherapy with Thrombin fibrin based glue and sclerosant (TFG/S . During the study period 18 patients were enrolled in the TGF/S group. 21 patients underwent variceal plugging with n - butyl cyanoacrylate (NBC . There was no significant difference in age/ sex , duration of bleed or time interval between onset of bleed and endotherapy. RESULTS: Patients undergoing endotherapy with TGF/S had less episodes of bleed , and greater eradication of varices. CONCLUSION: The results with thrombin / fibrin glue and sclerotherapy are highly encouraging. Well - designed trials need to be performed KEYWORDS:Gastric varices; Thrombin Sclerotherapy

  1. Thrombin contributes to bronchoalveolar lavage fluid mitogenicity in lung disease of the premature infant

    NARCIS (Netherlands)

    Dik, Willem A.; Zimmermann, Luc J. I.; Naber, Brigitta A.; Janssen, Daphne J.; van Kaam, Anton H. L. C.; Versnel, Marjan A.

    2003-01-01

    Chronic lung disease of prematurity (CLD) is a common consequence of neonatal respiratory distress syndrome (RDS) and is characterized by pulmonary fibrosis. Increased thrombin activity in the alveolar compartment is associated with pulmonary fibrosis in adults and animals, and contributes to

  2. Inactivation of single-chain urokinase-type plasminogen activator by thrombin in human subjects

    NARCIS (Netherlands)

    Braat, E. A.; Levi, M. [=Marcel M.; Bos, R.; Haverkate, F.; Lassen, M. R.; de Maat, M. P.; Rijken, D. C.

    1999-01-01

    Thrombin cleaves single-chain urokinase-type plasminogen activator (scu-PA) into a virtually inactive two-chain form (tcu-PA/T), a process that may protect a blood clot from early fibrinolysis. It is not known under what circumstances tcu-PA/T can be generated in vivo. We have studied the occurrence

  3. Astrid-2 EMMA Magnetic Calibration

    DEFF Research Database (Denmark)

    Merayo, José M.G.; Brauer, Peter; Risbo, Torben

    1998-01-01

    The Swedish micro-satellite Astrid-2 contains a tri-axial fluxgate magnetometer with the sensor co-located with a Technical University of Denmark (DTU) star camera for absolute attitude, and extended about 0.9 m on a hinged boom. The magnetometer is part of the RIT EMMA electric and magnetic fields...... experiment built as a collaboration between the DTU, Department of Automation and the Department of Plasma Physics, The Alfvenlaboratory, Royal Institute of Technology (RIT), Stockholm. The final magnetic calibration of the Astrid-2 satellite was done at the Lovoe Magnetic Observatory under the Geological...... Survey of Sweden near Stockholm on the night of May 15.-16., 1997. The magnetic calibration and the intercalibration between the star camera and the magnetic sensor was performed by measuring the Earth's magnetic field and simultaneously observing the star sky with the camera. The rotation matrix between...

  4. Thrombin induces epithelial-mesenchymal transition and collagen production by retinal pigment epithelial cells via autocrine PDGF-receptor signaling.

    Science.gov (United States)

    Bastiaans, Jeroen; van Meurs, Jan C; van Holten-Neelen, Conny; Nagtzaam, Nicole M A; van Hagen, P Martin; Chambers, Rachel C; Hooijkaas, Herbert; Dik, Willem A

    2013-12-19

    De-differentiation of RPE cells into mesenchymal cells (epithelial-mesenchymal transition; EMT) and associated collagen production contributes to development of proliferative vitreoretinopathy (PVR). In patients with PVR, intraocular coagulation cascade activation occurs and may play an important initiating role. Therefore, we examined the effect of the coagulation proteins factor Xa and thrombin on EMT and collagen production by RPE cells. Retinal pigment epithelial cells were stimulated with factor Xa or thrombin and the effect on zonula occludens (ZO)-1, α-smooth muscle actin (α-SMA), collagen, and platelet-derived growth factor (PDGF)-B were determined by real-time quantitative-polymerase chain reaction (RQ-PCR), immunofluorescence microscopy, and HPLC and ELISA for collagen and PDGF-BB in culture supernatants, respectively. PDGF-receptor activation was determined by phosphorylation analysis and inhibition studies using the PDGF-receptor tyrosine kinase inhibitor AG1296. Thrombin reduced ZO-1 gene expression (P production of α-SMA and collagen increased. In contrast to thrombin, factor Xa hardly stimulated EMT by RPE. Thrombin clearly induced PDGF-BB production and PDGF-Rβ chain phosphorylation in RPE. Moreover, AG1296 significantly blocked the effect of thrombin on EMT and collagen production. Our findings demonstrate that thrombin is a potent inducer of EMT by RPE via autocrine activation of PDGF-receptor signaling. Coagulation cascade-induced EMT of RPE may thus contribute to the formation of fibrotic retinal membranes in PVR and should be considered as treatment target in PVR.

  5. Ex vivo reversal of effects of rivaroxaban evaluated using thromboelastometry and thrombin generation assay

    Science.gov (United States)

    Schenk, B.; Würtinger, P.; Streif, W.; Sturm, W.; Fries, D.; Bachler, M.

    2016-01-01

    Background In major bleeding events, the new direct oral anticoagulants pose a great challenge for physicians. The aim of the study was to test for ex vivo reversal of the direct oral anticoagulant rivaroxaban with various non-specific reversal agents: prothrombin complex concentrate (PCC), activated prothrombin complex concentrate (aPCC), recombinant activated factor VII (rFVIIa), and fibrinogen concentrate (FI). Methods Blood was obtained from healthy volunteers and from patients treated with rivaroxaban. Blood samples from healthy volunteers were spiked with rivaroxaban to test the correlation between rivaroxaban concentration and coagulation tests. Patient blood samples were spiked with various concentrations of the above-mentioned agents and analysed using thromboelastometry and thrombin generation. Results When added in vitro, rivaroxaban was significantly (P<0.05) correlated with ROTEM® thromboelastometry EXTEM (extrinsic coagulation pathway) clotting time (CT), time to maximal velocity (MaxV−t), and with all measured thrombin generation parameters. In vivo, CT, MaxV−t, lag time, and peak thrombin generation (Cmax) were significantly correlated with rivaroxaban concentrations. Regarding reversal of rivaroxaban, all tested agents significantly (P<0.05) reduced EXTEM CT, but to different extents: rFVIIa by 68%, aPCC by 47%, PCC by 17%, and FI by 9%. Only rFVIIa reversed EXTEM CT to baseline values. Both PCC (+102%) and aPCC (+232%) altered overall thrombin generation (area under the curve) and increased Cmax (+461% for PCC, +87.5% for aPCC). Conclusions Thromboelastometry and thrombin generation assays do not favour the same reversal agents for rivaroxaban anticoagulation. Controlled clinical trials are urgently needed to establish doses and clinical efficacy of potential reversal agents. Clinical trial registration EudracCT trial no. 213-00474-30. PMID:27623677

  6. Effects of fibrin, thrombin, and blood on breast capsule formation in a preclinical model.

    Science.gov (United States)

    Marques, Marisa; Brown, Spencer A; Cordeiro, Natália D S; Rodrigues-Pereira, Pedro; Cobrado, M Luís; Morales-Helguera, Aliuska; Lima, Nuno; Luís, André; Mendanha, Mário; Gonçalves-Rodrigues, Acácio; Amarante, José

    2011-03-01

    The root cause of capsular contracture (CC) associated with breast implants is unknown. Recent evidence points to the possible role of fibrin and bacteria in CC formation. The authors sought to determine whether fibrin, thrombin, and blood modulated the histological and microbiological outcomes of breast implant capsule formation in a rabbit model. The authors carried out a case-control study to assess the influence of fibrin, thrombin, and blood on capsule wound healing in a rabbit model. Eighteen New Zealand white rabbits received four tissue expanders. One expander acted as a control, whereas the other expander pockets received one of the following: fibrin glue, rabbit blood, or thrombin sealant. Intracapsular pressure/volume curves were compared among the groups, and histological and microbiological evaluations were performed (capsules, tissue expanders, rabbit skin, and air). The rabbits were euthanized at two or four weeks. At four weeks, the fibrin and thrombin expanders demonstrated significantly decreased intracapsular pressure compared to the control group. In the control and fibrin groups, mixed inflammation correlated with decreased intracapsular pressure, whereas mononuclear inflammation correlated with increased intracapsular pressure. The predominant isolate in the capsules, tissue expanders, and rabbit skin was coagulase-negative staphylococci. For fibrin and thrombin, both cultures that showed an organism other than staphylococci and cultures that were negative were associated with decreased intracapsular pressure, whereas cultures positive for staphylococci were associated with increased intracapsular pressure. Fibrin application during breast implantation may reduce rates of CC, but the presence of staphylococci is associated with increased capsule pressure even in the presence of fibrin, so care should be taken to avoid bacterial contamination.

  7. Salmon and human thrombin differentially regulate radicular pain, glial-induced inflammation and spinal neuronal excitability through protease-activated receptor-1.

    Directory of Open Access Journals (Sweden)

    Jenell R Smith

    Full Text Available Chronic neck pain is a major problem with common causes including disc herniation and spondylosis that compress the spinal nerve roots. Cervical nerve root compression in the rat produces sustained behavioral hypersensitivity, due in part to the early upregulation of pro-inflammatory cytokines, the sustained hyperexcitability of neurons in the spinal cord and degeneration in the injured nerve root. Through its activation of the protease-activated receptor-1 (PAR1, mammalian thrombin can enhance pain and inflammation; yet at lower concentrations it is also capable of transiently attenuating pain which suggests that PAR1 activation rate may affect pain maintenance. Interestingly, salmon-derived fibrin, which contains salmon thrombin, attenuates nerve root-induced pain and inflammation, but the mechanisms of action leading to its analgesia are unknown. This study evaluates the effects of salmon thrombin on nerve root-mediated pain, axonal degeneration in the root, spinal neuronal hyperexcitability and inflammation compared to its human counterpart in the context of their enzymatic capabilities towards coagulation substrates and PAR1. Salmon thrombin significantly reduces behavioral sensitivity, preserves neuronal myelination, reduces macrophage infiltration in the injured nerve root and significantly decreases spinal neuronal hyperexcitability after painful root compression in the rat; whereas human thrombin has no effect. Unlike salmon thrombin, human thrombin upregulates the transcription of IL-1β and TNF-α and the secretion of IL-6 by cortical cultures. Salmon and human thrombins cleave human fibrinogen-derived peptides and form clots with fibrinogen with similar enzymatic activities, but salmon thrombin retains a higher enzymatic activity towards coagulation substrates in the presence of antithrombin III and hirudin compared to human thrombin. Conversely, salmon thrombin activates a PAR1-derived peptide more weakly than human thrombin. These

  8. Toward designing for trust in database automation

    Energy Technology Data Exchange (ETDEWEB)

    Duez, P. P.; Jamieson, G. A. [Cognitive Engineering Laboratory, Univ. of Toronto, 5 King' s College Rd., Toronto, Ont. M5S 3G8 (Canada)

    2006-07-01

    Appropriate reliance on system automation is imperative for safe and productive work, especially in safety-critical systems. It is unsafe to rely on automation beyond its designed use; conversely, it can be both unproductive and unsafe to manually perform tasks that are better relegated to automated tools. Operator trust in automated tools mediates reliance, and trust appears to affect how operators use technology. As automated agents become more complex, the question of trust in automation is increasingly important. In order to achieve proper use of automation, we must engender an appropriate degree of trust that is sensitive to changes in operating functions and context. In this paper, we present research concerning trust in automation in the domain of automated tools for relational databases. Lee and See have provided models of trust in automation. One model developed by Lee and See identifies three key categories of information about the automation that lie along a continuum of attributional abstraction. Purpose-, process-and performance-related information serve, both individually and through inferences between them, to describe automation in such a way as to engender r properly-calibrated trust. Thus, one can look at information from different levels of attributional abstraction as a general requirements analysis for information key to appropriate trust in automation. The model of information necessary to engender appropriate trust in automation [1] is a general one. Although it describes categories of information, it does not provide insight on how to determine the specific information elements required for a given automated tool. We have applied the Abstraction Hierarchy (AH) to this problem in the domain of relational databases. The AH serves as a formal description of the automation at several levels of abstraction, ranging from a very abstract purpose-oriented description to a more concrete description of the resources involved in the automated process

  9. Toward designing for trust in database automation

    International Nuclear Information System (INIS)

    Duez, P. P.; Jamieson, G. A.

    2006-01-01

    Appropriate reliance on system automation is imperative for safe and productive work, especially in safety-critical systems. It is unsafe to rely on automation beyond its designed use; conversely, it can be both unproductive and unsafe to manually perform tasks that are better relegated to automated tools. Operator trust in automated tools mediates reliance, and trust appears to affect how operators use technology. As automated agents become more complex, the question of trust in automation is increasingly important. In order to achieve proper use of automation, we must engender an appropriate degree of trust that is sensitive to changes in operating functions and context. In this paper, we present research concerning trust in automation in the domain of automated tools for relational databases. Lee and See have provided models of trust in automation. One model developed by Lee and See identifies three key categories of information about the automation that lie along a continuum of attributional abstraction. Purpose-, process-and performance-related information serve, both individually and through inferences between them, to describe automation in such a way as to engender r properly-calibrated trust. Thus, one can look at information from different levels of attributional abstraction as a general requirements analysis for information key to appropriate trust in automation. The model of information necessary to engender appropriate trust in automation [1] is a general one. Although it describes categories of information, it does not provide insight on how to determine the specific information elements required for a given automated tool. We have applied the Abstraction Hierarchy (AH) to this problem in the domain of relational databases. The AH serves as a formal description of the automation at several levels of abstraction, ranging from a very abstract purpose-oriented description to a more concrete description of the resources involved in the automated process

  10. Process automation

    International Nuclear Information System (INIS)

    Moser, D.R.

    1986-01-01

    Process automation technology has been pursued in the chemical processing industries and to a very limited extent in nuclear fuel reprocessing. Its effective use has been restricted in the past by the lack of diverse and reliable process instrumentation and the unavailability of sophisticated software designed for process control. The Integrated Equipment Test (IET) facility was developed by the Consolidated Fuel Reprocessing Program (CFRP) in part to demonstrate new concepts for control of advanced nuclear fuel reprocessing plants. A demonstration of fuel reprocessing equipment automation using advanced instrumentation and a modern, microprocessor-based control system is nearing completion in the facility. This facility provides for the synergistic testing of all chemical process features of a prototypical fuel reprocessing plant that can be attained with unirradiated uranium-bearing feed materials. The unique equipment and mission of the IET facility make it an ideal test bed for automation studies. This effort will provide for the demonstration of the plant automation concept and for the development of techniques for similar applications in a full-scale plant. A set of preliminary recommendations for implementing process automation has been compiled. Some of these concepts are not generally recognized or accepted. The automation work now under way in the IET facility should be useful to others in helping avoid costly mistakes because of the underutilization or misapplication of process automation. 6 figs

  11. Parameterizations for reducing camera reprojection error for robot-world hand-eye calibration

    Science.gov (United States)

    Accurate robot-world, hand-eye calibration is crucial to automation tasks. In this paper, we discuss the robot-world, hand-eye calibration problem which has been modeled as the linear relationship AX equals ZB, where X and Z are the unknown calibration matrices composed of rotation and translation ...

  12. Factor Xa generation by computational modeling: an additional discriminator to thrombin generation evaluation.

    Directory of Open Access Journals (Sweden)

    Kathleen E Brummel-Ziedins

    Full Text Available Factor (fXa is a critical enzyme in blood coagulation that is responsible for the initiation and propagation of thrombin generation. Previously we have shown that analysis of computationally generated thrombin profiles is a tool to investigate hemostasis in various populations. In this study, we evaluate the potential of computationally derived time courses of fXa generation as another approach for investigating thrombotic risk. Utilizing the case (n = 473 and control (n = 426 population from the Leiden Thrombophilia Study and each individual's plasma protein factor composition for fII, fV, fVII, fVIII, fIX, fX, antithrombin and tissue factor pathway inhibitor, tissue factor-initiated total active fXa generation was assessed using a mathematical model. FXa generation was evaluated by the area under the curve (AUC, the maximum rate (MaxR and level (MaxL and the time to reach these, TMaxR and TMaxL, respectively. FXa generation was analyzed in the entire populations and in defined subgroups (by sex, age, body mass index, oral contraceptive use. The maximum rates and levels of fXa generation occur over a 10- to 12- fold range in both cases and controls. This variation is larger than that observed with thrombin (3-6 fold in the same population. The greatest risk association was obtained using either MaxR or MaxL of fXa generation; with an ∼2.2 fold increased risk for individuals exceeding the 90(th percentile. This risk was similar to that of thrombin generation(MaxR OR 2.6. Grouping defined by oral contraceptive (OC use in the control population showed the biggest differences in fXa generation; a >60% increase in the MaxR upon OC use. FXa generation can distinguish between a subset of individuals characterized by overlapping thrombin generation profiles. Analysis of fXa generation is a phenotypic characteristic which may prove to be a more sensitive discriminator than thrombin generation among all individuals.

  13. Thrombin and factor Xa link the coagulation system with liver fibrosis.

    Science.gov (United States)

    Dhar, Ameet; Sadiq, Fouzia; Anstee, Quentin M; Levene, Adam P; Goldin, Robert D; Thursz, Mark R

    2018-05-08

    Thrombin activates hepatic stellate cells via protease-activated receptor-1. The role of Factor Xa (FXa) in hepatic fibrosis has not been elucidated. We aimed to evaluate the impact of FXa and thrombin in vitro on stellate cells and their respective inhibition in vivo using a rodent model of hepatic fibrosis. HSC-LX2 cells were incubated with FXa and/or thrombin in cell culture, stained for αSMA and relative gene expression and gel contraction calculated. C57BL/6 J mice were administered thioacetamide (TAA) for 8 weeks with Rivaroxaban (n = 15) or Dabigatran (n = 15). Control animals received TAA alone (n = 15). Fibrosis was scored and quantified using digital image analysis and hepatic tissue hydroxyproline estimated. Stellate cells treated with FXa and thrombin demonstrated upregulation of procollagen, TGF-beta, αSMA and significant cell contraction (43.48%+/- 4.12) compared to culturing with FXa or thrombin alone (26.90%+/- 8.90, p = 0.02; 13.1%+/- 9.84, p < 0.001). Mean fibrosis score, percentage area of fibrosis and hepatic hydroxyproline content (2.46 vs 4.08, p = 0.008; 2.02% vs 3.76%, p = 0.012; 276.0 vs 651.3, p = 0.0001) were significantly reduced in mice treated with the FXa inhibitor compared to control mice. FXa inhibition was significantly more effective than thrombin inhibition in reducing percentage area of fibrosis and hepatic hydroxyproline content (2.02% vs 3.70%,p = 0.031; 276.0 vs 413.1,p = 0.001). FXa promotes stellate cell contractility and activation. Early inhibition of coagulation using a FXa inhibitor significantly reduces TAA induced murine liver fibrosis and may be a viable treatment for liver fibrosis in patients.

  14. Selective albumin-binding surfaces modified with a thrombin-inhibiting peptide.

    Science.gov (United States)

    Freitas, Sidónio C; Maia, Sílvia; Figueiredo, Ana C; Gomes, Paula; Pereira, Pedro J B; Barbosa, Mário A; Martins, M Cristina L

    2014-03-01

    Blood-contacting medical devices have been associated with severe clinical complications, such as thrombus formation, triggered by the activation of the coagulation cascade due to the adsorption of certain plasma proteins on the surface of biomaterials. Hence, the coating of such surfaces with antithrombotic agents has been used to increase biomaterial haemocompatibility. Biomaterial-induced clotting may also be decreased by albumin adsorption from blood plasma in a selective and reversible way, since this protein is not involved in the coagulation cascade. In this context, this paper reports that the immobilization of the thrombin inhibitor D-Phe-Pro-D-Arg-D-Thr-CONH2 (fPrt) onto nanostructured surfaces induces selective and reversible adsorption of albumin, delaying the clotting time when compared to peptide-free surfaces. fPrt, synthesized with two glycine residues attached to the N-terminus (GGfPrt), was covalently immobilized onto self-assembled monolayers (SAMs) having different ratios of carboxylate-hexa(ethylene glycol)- and tri(ethylene glycol)-terminated thiols (EG6-COOH/EG3) that were specifically designed to control GGfPrt orientation, exposure and density at the molecular level. In solution, GGfPrt was able to inactivate the enzymatic activity of thrombin and to delay plasma clotting time in a concentration-dependent way. After surface immobilization, and independently of its concentration, GGfPrt lost its selectivity to thrombin and its capacity to inhibit thrombin enzymatic activity against the chromogenic substrate n-p-tosyl-Gly-Pro-Arg-p-nitroanilide. Nevertheless, surfaces with low concentrations of GGfPrt could delay the capacity of adsorbed thrombin to cleave fibrinogen. In contrast, GGfPrt immobilized in high concentrations was found to induce the procoagulant activity of the adsorbed thrombin. However, all surfaces containing GGfPrt have a plasma clotting time similar to the negative control (empty polystyrene wells), showing resistance to

  15. Turn-on fluorescence sensor based on single-walled-carbon-nanohorn-peptide complex for the detection of thrombin.

    Science.gov (United States)

    Zhu, Shuyun; Liu, Zhongyuan; Hu, Lianzhe; Yuan, Yali; Xu, Guobao

    2012-12-14

    Proteases play a central role in several widespread diseases. Thus, there is a great need for the fast and sensitive detection of various proteolytic enzymes. Herein, we have developed a carbon nanotube (CNT)-based protease biosensing platform that uses peptides as a fluorescence probe for the first time. Single-walled carbon nanohorns (SWCNHs) and thrombin were used to demonstrate this detection strategy. SWCNHs can adsorb a fluorescein-based dye (FAM)-labeled peptide (FAM-pep) and quench the fluorescence of FAM. In contrast, thrombin can cleave FAM-pep on SWCNHs and recover the fluorescence of FAM, which allows the sensitive detection of thrombin. This biosensor has a high sensitivity and selectivity toward thrombin, with a detection limit of 100 pM. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  16. A functional single nucleotide polymorphism in the thrombin-activatable fibrinolysis inhibitor (TAFI) gene associates with outcome of meningococcal disease

    NARCIS (Netherlands)

    Kremer Hovinga, J. A.; Franco, R. F.; Zago, M. A.; ten Cate, Hugo; Westendorp, R. G. J.; Reitsma, P. H.

    2004-01-01

    In meningococcal sepsis, disseminated intravascular coagulation with deposition of fibrin and formation of microthrombi occurs in various organs and enhanced inhibition of fibrinolysis is associated with adverse outcome. Recently, TAFI (thrombin-activatable fibrinolysis inhibitor) was identified as

  17. Direct electrochemistry and electrocatalysis of a glucose oxidase-functionalized bioconjugate as a trace label for ultrasensitive detection of thrombin.

    Science.gov (United States)

    Bai, Lijuan; Yuan, Ruo; Chai, Yaqin; Yuan, Yali; Wang, Yan; Xie, Shunbi

    2012-11-18

    For the first time, a glucose oxidase-functionalized bioconjugate was prepared and served as a new trace label through its direct electrochemistry and electrocatalysis in a sandwich-type electrochemical aptasensor for ultrasensitive detection of thrombin.

  18. Binding of α2-macroglobulin-thrombin complexes and methylamine-treated α2-macroglobulin to human blood monocytes

    International Nuclear Information System (INIS)

    Straight, D.L.; Jakoi, L.; McKee, P.A.; Snyderman, R.

    1988-01-01

    The binding of α 2 -macroglobulin (α 2 M) to human peripheral blood monocytes was investigated. Monocytes, the precursors of tissue macrophages, were isolated from fresh blood by centrifugal elutriation or density gradient centrifugation. Binding studies were performed using 125 I-labeled α 2 M. Cells and bound ligand were separated from free ligand by rapid vacuum filtration. Nonlinear least-squares analysis of data obtained in direct binding studies at 0 0 C showed that monocytes bound the α 2 M-thrombin complex with a K/sub d/ 3.0 +- .09 nM and the monocyte had 1545 +- 153 sitescell. Thrombin alone did not compete for the site. Binding was divalent cation dependent. Direct binding studies also demonstrated that monocytes bound methylamine-treated α 2 M in a manner similar to α 2 M-thrombin. Competitive binding studies showed that α 2 M-thrombin and methylamine-treated α 2 M bound to the same sites on the monocyte. In contrast, native α 2 M did not compete with α 2 M-thrombin for the site. Studies done at 37 0 C suggested that after binding, the monocyte internalized and degraded α 2 M-thrombin and excreted the degradation products. Receptor turnover and degradation of α 2 M-thrombin complexes were blocked in monocytes treated with chloroquine, an inhibitor of lysosomal function. The results indicate that human monocytes have a divalent cation dependent, high-affinity binding site for α 2 M-thrombin and methylamine-treated α 2 M which may function to clear α 2 M-proteinase complexes from the circulation

  19. Plasmid-Mediated Resistance to Thrombin-Induced Platelet Microbicidal Protein in Staphylococci: Role of the qacA Locus

    OpenAIRE

    Kupferwasser, Leon Iri; Skurray, Ronald A.; Brown, Melissa H.; Firth, Neville; Yeaman, Michael R.; Bayer, Arnold S.

    1999-01-01

    Thrombin-induced platelet microbicidal protein 1 (tPMP-1) is a small, cationic peptide released from rabbit platelets following thrombin stimulation. In vitro resistance to this peptide among strains of Staphylococcus aureus correlates with the survival advantage of such strains at sites of endothelial damage in humans as well as in experimental endovascular infections. The mechanisms involved in the phenotypic resistance of S. aureus to tPMP-1 are not fully delineated. The plasmid-encoded st...

  20. Automated analysis of slitless spectra. II. Quasars

    International Nuclear Information System (INIS)

    Edwards, G.; Beauchemin, M.; Borra, F.

    1988-01-01

    Automated software have been developed to process slitless spectra. The software, described in a previous paper, automatically separates stars from extended objects and quasars from stars. This paper describes the quasar search techniques and discusses the results. The performance of the software is compared and calibrated with a plate taken in a region of SA 57 that has been extensively surveyed by others using a variety of techniques: the proposed automated software performs very well. It is found that an eye search of the same plate is less complete than the automated search: surveys that rely on eye searches suffer from incompleteness at least from a magnitude brighter than the plate limit. It is shown how the complete automated analysis of a plate and computer simulations are used to calibrate and understand the characteristics of the present data. 20 references

  1. Distribution automation

    International Nuclear Information System (INIS)

    Gruenemeyer, D.

    1991-01-01

    This paper reports on a Distribution Automation (DA) System enhances the efficiency and productivity of a utility. It also provides intangible benefits such as improved public image and market advantages. A utility should evaluate the benefits and costs of such a system before committing funds. The expenditure for distribution automation is economical when justified by the deferral of a capacity increase, a decrease in peak power demand, or a reduction in O and M requirements

  2. Large-scale preparation of active caspase-3 in E. coli by designing its thrombin-activatable precursors

    Directory of Open Access Journals (Sweden)

    Park Sung

    2008-12-01

    Full Text Available Abstract Background Caspase-3, a principal apoptotic effector that cleaves the majority of cellular substrates, is an important medicinal target for the treatment of cancers and neurodegenerative diseases. Large amounts of the protein are required for drug discovery research. However, previous efforts to express the full-length caspase-3 gene in E. coli have been unsuccessful. Results Overproducers of thrombin-activatable full-length caspase-3 precursors were prepared by engineering the auto-activation sites of caspase-3 precursor into a sequence susceptible to thrombin hydrolysis. The engineered precursors were highly expressed as soluble proteins in E. coli and easily purified by affinity chromatography, to levels of 10–15 mg from 1 L of E. coli culture, and readily activated by thrombin digestion. Kinetic evaluation disclosed that thrombin digestion enhanced catalytic activity (kcat/KM of the precursor proteins by two orders of magnitude. Conclusion A novel method for a large-scale preparation of active caspase-3 was developed by a strategic engineering to lack auto-activation during expression with amino acid sequences susceptible to thrombin, facilitating high-level expression in E. coli. The precursor protein was easily purified and activated through specific cleavage at the engineered sites by thrombin, generating active caspase-3 in high yields.

  3. Thrombin effectuates therapeutic arteriogenesis in the rabbit hindlimb ischemia model: A quantitative analysis by computerized in vivo imaging

    International Nuclear Information System (INIS)

    Kagadis, George C.; Karnabatidis, Dimitrios; Katsanos, Konstantinos; Diamantopoulos, Athanassios; Samaras, Nikolaos; Maroulis, John; Siablis, Dimitrios; Nikiforidis, George C.

    2006-01-01

    We report on an experimental mammalian controlled study that documents arteriogenic capacity of thrombin and utilizes computerized algorithms to quantify the newly formed vessels. Hindlimb ischemia was surgically invoked in 10 New Zealand white rabbits. After quiescence of endogenous angiogenesis heterologous bovine thrombin was intramuscularly injected (1500 units) in one hindlimb per rabbit (Group T). Contralateral limbs were infused with normal saline (Group C). Digital subtraction angiography (DSA) of both limbs was performed after thrombin infusion by selective cannulation of the abdominal aorta and digital images were post-processed with computerized algorithms in order to enhance newly formed vessels. Total vessel area and total vessel length were quantified. In vivo functional evaluation included measurements of blood flow volume at the level of the external iliac artery by Doppler ultrasonography both at baseline and at 20 days after thrombin infusion. Total vessel area and length (in pixels) were 14,713+/-1023 and 5466+/-1327 in group T versus 12,015+/-2557 and 4598+/-1269 in group C (p=0.0062 and 0.1526, respectively). Blood flow volumes (ml/min) at baseline and at 20 days after thrombin infusion were 25.87+/-11.09 and 38.06+/-11.72 in group T versus 26.57+/-11.19 and 20.35+/-7.20 in group C (p=0.8898 and 0.0007, respectively). Intramuscular thrombin effectuates an arteriogenic response in the rabbit hindlimb ischemia model. Computerized algorithms may enable accurate quantification of the neovascularization outcome

  4. Thermodynamic compensation upon binding to exosite 1 and the active site of thrombin.

    Science.gov (United States)

    Treuheit, Nicholas A; Beach, Muneera A; Komives, Elizabeth A

    2011-05-31

    Several lines of experimental evidence including amide exchange and NMR suggest that ligands binding to thrombin cause reduced backbone dynamics. Binding of the covalent inhibitor dPhe-Pro-Arg chloromethyl ketone to the active site serine, as well as noncovalent binding of a fragment of the regulatory protein, thrombomodulin, to exosite 1 on the back side of the thrombin molecule both cause reduced dynamics. However, the reduced dynamics do not appear to be accompanied by significant conformational changes. In addition, binding of ligands to the active site does not change the affinity of thrombomodulin fragments binding to exosite 1; however, the thermodynamic coupling between exosite 1 and the active site has not been fully explored. We present isothermal titration calorimetry experiments that probe changes in enthalpy and entropy upon formation of binary ligand complexes. The approach relies on stringent thrombin preparation methods and on the use of dansyl-l-arginine-(3-methyl-1,5-pantanediyl)amide and a DNA aptamer as ligands with ideal thermodynamic signatures for binding to the active site and to exosite 1. Using this approach, the binding thermodynamic signatures of each ligand alone as well as the binding signatures of each ligand when the other binding site was occupied were measured. Different exosite 1 ligands with widely varied thermodynamic signatures cause a similar reduction in ΔH and a concomitantly lower entropy cost upon DAPA binding at the active site. The results suggest a general phenomenon of enthalpy-entropy compensation consistent with reduction of dynamics/increased folding of thrombin upon ligand binding to either the active site or exosite 1.

  5. Spontaneous pseudoaneurysm of the uterine artery during pregnancy treated by direct thrombin injection: A case report

    Energy Technology Data Exchange (ETDEWEB)

    Hong, Jung Hee; Kim, See Hyung; Kim, Young Hwan [Dept. Radiology, Keimyung University School of Medicine, Dongsan Medical Center, Daegu (Korea, Republic of)

    2016-04-15

    Pseudoaneurysm of uterine artery during pregnancy is a very rare disease. It is mostly associated with uterine artery injury, usually occurring after proceeding conditions such as history of gynecologic operation and infection. However, the best treatment modality has not been established yet. Herein, we reported a case of spontaneous formation of uterine artery pseudoaneurysm during pregnancy treated by direct thrombin injection without any complication or recurrence.

  6. Spontaneous pseudoaneurysm of the uterine artery during pregnancy treated by direct thrombin injection: A case report

    International Nuclear Information System (INIS)

    Hong, Jung Hee; Kim, See Hyung; Kim, Young Hwan

    2016-01-01

    Pseudoaneurysm of uterine artery during pregnancy is a very rare disease. It is mostly associated with uterine artery injury, usually occurring after proceeding conditions such as history of gynecologic operation and infection. However, the best treatment modality has not been established yet. Herein, we reported a case of spontaneous formation of uterine artery pseudoaneurysm during pregnancy treated by direct thrombin injection without any complication or recurrence

  7. First Steps in the Direction of Synthetic, Allosteric, Direct Inhibitors of Thrombin and Factor Xa

    OpenAIRE

    Verghese, Jenson; Liang, Aiye; Sidhu, Preet Pal Singh; Hindle, Michael; Zhou, Qibing; Desai, Umesh R.

    2009-01-01

    Designing non-saccharide functional mimics of heparin is a major challenge. In this work, a library of small, aromatic molecules based on the sulfated DHP scaffold was synthesized and screened against thrombin and factor Xa. The results reveal that i) selected monomeric benzofuran derivatives inhibit the two enzymes, albeit weakly; ii) the two enzymes recognize different structural features in the benzofurans studied suggesting significant selectivity of recognition; and iii) the mechanism of...

  8. Successful endovascular treatment of a hemodialysis graft pseudoaneurysm by covered stent and direct percutaneous thrombin injection.

    LENUS (Irish Health Repository)

    Keeling, Aoife N

    2011-07-25

    Vascular access for hemodialysis remains a challenge for nephrologists, vascular surgeons, and interventional radiologists alike. Arteriovenous fistula and synthetic grafts remain the access of choice for long-term hemodialysis; however, they are subject to complications from infection and repeated needle cannulation. Pseudoaneurysms are an increasingly recognized adverse event. At present, there are many minimally invasive methods to repair these wall defects. We present a graft pseudoaneurysm, which required a combination of endovascular stent graft placement and percutaneous thrombin injection for successful occlusion.

  9. Automated image quality assessment for chest CT scans.

    Science.gov (United States)

    Reeves, Anthony P; Xie, Yiting; Liu, Shuang

    2018-02-01

    Medical image quality needs to be maintained at standards sufficient for effective clinical reading. Automated computer analytic methods may be applied to medical images for quality assessment. For chest CT scans in a lung cancer screening context, an automated quality assessment method is presented that characterizes image noise and image intensity calibration. This is achieved by image measurements in three automatically segmented homogeneous regions of the scan: external air, trachea lumen air, and descending aorta blood. Profiles of CT scanner behavior are also computed. The method has been evaluated on both phantom and real low-dose chest CT scans and results show that repeatable noise and calibration measures may be realized by automated computer algorithms. Noise and calibration profiles show relevant differences between different scanners and protocols. Automated image quality assessment may be useful for quality control for lung cancer screening and may enable performance improvements to automated computer analysis methods. © 2017 American Association of Physicists in Medicine.

  10. Structure-activity studies and therapeutic potential of host defense peptides of human thrombin.

    Science.gov (United States)

    Kasetty, Gopinath; Papareddy, Praveen; Kalle, Martina; Rydengård, Victoria; Mörgelin, Matthias; Albiger, Barbara; Malmsten, Martin; Schmidtchen, Artur

    2011-06-01

    Peptides of the C-terminal region of human thrombin are released upon proteolysis and identified in human wounds. In this study, we wanted to investigate minimal determinants, as well as structural features, governing the antimicrobial and immunomodulating activity of this peptide region. Sequential amino acid deletions of the peptide GKYGFYTHVFRLKKWIQKVIDQFGE (GKY25), as well as substitutions at strategic and structurally relevant positions, were followed by analyses of antimicrobial activity against the Gram-negative bacteria Escherichia coli and Pseudomonas aeruginosa, the Gram-positive bacterium Staphylococcus aureus, and the fungus Candida albicans. Furthermore, peptide effects on lipopolysaccharide (LPS)-, lipoteichoic acid-, or zymosan-induced macrophage activation were studied. The thrombin-derived peptides displayed length- and sequence-dependent antimicrobial as well as immunomodulating effects. A peptide length of at least 20 amino acids was required for effective anti-inflammatory effects in macrophage models, as well as optimal antimicrobial activity as judged by MIC assays. However, shorter (>12 amino acids) variants also displayed significant antimicrobial effects. A central K14 residue was important for optimal antimicrobial activity. Finally, one peptide variant, GKYGFYTHVFRLKKWIQKVI (GKY20) exhibiting improved selectivity, i.e., low toxicity and a preserved antimicrobial as well as anti-inflammatory effect, showed efficiency in mouse models of LPS shock and P. aeruginosa sepsis. The work defines structure-activity relationships of C-terminal host defense peptides of thrombin and delineates a strategy for selecting peptide epitopes of therapeutic interest.

  11. Molecular modeling studies of novel retro-binding tripeptide active-site inhibitors of thrombin.

    Science.gov (United States)

    Lau, W F; Tabernero, L; Sack, J S; Iwanowicz, E J

    1995-08-01

    A novel series of retro-binding tripeptide thrombin active-site inhibitors was recently developed (Iwanowicz, E. I. et al. J. Med. Chem. 1994, 37, 2111(1)). It was hypothesized that the binding mode for these inhibitors is similar to that of the first three N-terminal residues of hirudin. This binding hypothesis was subsequently verified when the crystal structure of a member of this series, BMS-183,507 (N-[N-[N-[4-(Aminoiminomethyl)amino[-1-oxobutyl]-L- phenylalanyl]-L-allo-threonyl]-L-phenylalanine, methyl ester), was determined (Taberno, L.J. Mol. Biol. 1995, 246, 14). The methodology for developing the binding models of these inhibitors, the structure-activity relationships (SAR) and modeling studies that led to the elucidation of the proposed binding mode is described. The crystal structure of BMS-183,507/human alpha-thrombin is compared with the crystal structure of hirudin/human alpha-thrombin (Rydel, T.J. et al. Science 1990, 249,227; Rydel, T.J. et al. J. Mol Biol. 1991, 221, 583; Grutter, M.G. et al. EMBO J. 1990, 9, 2361) and with the computational binding model of BMS-183,507.

  12. Elevated Cytokines, Thrombin and PAI-1 in Severe HCPS Patients Due to Sin Nombre Virus

    Directory of Open Access Journals (Sweden)

    Virginie Bondu

    2015-02-01

    Full Text Available Sin Nombre Hantavirus (SNV, Bunyaviridae Hantavirus is a Category A pathogen that causes Hantavirus Cardiopulmonary Syndrome (HCPS with case fatality ratios generally ranging from 30% to 50%. HCPS is characterized by vascular leakage due to dysregulation of the endothelial barrier function. The loss of vascular integrity results in non-cardiogenic pulmonary edema, shock, multi-organ failure and death. Using Electric Cell-substrate Impedance Sensing (ECIS measurements, we found that plasma samples drawn from University of New Mexico Hospital patients with serologically-confirmed HCPS, induce loss of cell-cell adhesion in confluent epithelial and endothelial cell monolayers grown in ECIS cultureware. We show that the loss of cell-cell adhesion is sensitive to both thrombin and plasmin inhibitors in mild cases, and to thrombin only inhibition in severe cases, suggesting an increasing prothrombotic state with disease severity. A proteomic profile (2D gel electrophoresis and mass spectrometry of HCPS plasma samples in our cohort revealed robust antifibrinolytic activity among terminal case patients. The prothrombotic activity is highlighted by acute ≥30 to >100 fold increases in active plasminogen activator inhibitor (PAI-1 which, preceded death of the subjects within 48 h. Taken together, this suggests that PAI-1 might be a response to the severe pathology as it is expected to reduce plasmin activity and possibly thrombin activity in the terminal patients.

  13. Virtual automation.

    Science.gov (United States)

    Casis, E; Garrido, A; Uranga, B; Vives, A; Zufiaurre, C

    2001-01-01

    Total laboratory automation (TLA) can be substituted in mid-size laboratories by a computer sample workflow control (virtual automation). Such a solution has been implemented in our laboratory using PSM, software developed in cooperation with Roche Diagnostics (Barcelona, Spain), to this purpose. This software is connected to the online analyzers and to the laboratory information system and is able to control and direct the samples working as an intermediate station. The only difference with TLA is the replacement of transport belts by personnel of the laboratory. The implementation of this virtual automation system has allowed us the achievement of the main advantages of TLA: workload increase (64%) with reduction in the cost per test (43%), significant reduction in the number of biochemistry primary tubes (from 8 to 2), less aliquoting (from 600 to 100 samples/day), automation of functional testing, drastic reduction of preanalytical errors (from 11.7 to 0.4% of the tubes) and better total response time for both inpatients (from up to 48 hours to up to 4 hours) and outpatients (from up to 10 days to up to 48 hours). As an additional advantage, virtual automation could be implemented without hardware investment and significant headcount reduction (15% in our lab).

  14. Mechanics of log calibration

    International Nuclear Information System (INIS)

    Waller, W.C.; Cram, M.E.; Hall, J.E.

    1975-01-01

    For any measurement to have meaning, it must be related to generally accepted standard units by a valid and specified system of comparison. To calibrate well-logging tools, sensing systems are designed which produce consistent and repeatable indications over the range for which the tool was intended. The basics of calibration theory, procedures, and calibration record presentations are reviewed. Calibrations for induction, electrical, radioactivity, and sonic logging tools will be discussed. The authors' intent is to provide an understanding of the sources of errors, of the way errors are minimized in the calibration process, and of the significance of changes in recorded calibration data

  15. A Pseudoaneurysm of the Deep Palmar Arch After Penetrating Trauma to the Hand: Successful Exclusion by Ultrasound Guided Percutaneous Thrombin Injection

    Directory of Open Access Journals (Sweden)

    A. Bosman

    Full Text Available : Introduction: Pseudoaneurysm of the hand is a rare condition; most are treated surgically. Ultrasound guided thrombin injection has not previously been reported as a treatment option for pseudoaneurysms of the deep palmar arch. Report: A man was referred to the emergency department with a swollen, painful hand after penetrating trauma. On physical examination, a pulsating tumor was found on the dorsum of the hand. Imaging revealed a pseudoaneurysm vascularized by the deep palmar arch. Ultrasound guided percutaneous thrombin injection was successfully performed. Conclusion: Thrombin injection might be a safe alternative option in the treatment of pseudoaneurysm of the deep palmar arch. Keywords: Deep palmar arch, Pseudoaneurysm, Thrombin injection

  16. Protein C inhibits endocytosis of thrombin-thrombomodulin complexes in A549 lung cancer cells and human umbilical vein endothelial cells

    International Nuclear Information System (INIS)

    Maruyama, I.; Majerus, P.W.

    1987-01-01

    We investigated the effect of protein C on the endocytosis of thrombin-thrombomodulin complexes. We previously showed that exposure of umbilical vein endothelial cells to thrombin stimulated the internalization and degradation of thrombin. A similar internalization was stimulated by a monoclonal antithrombomodulin antibody. We have repeated these studies in the presence of protein C and found that endocytosis of 125 I-thrombin-thrombomodulin complexes, but not 125 I-antithrombomodulin-thrombomodulin complexes, is inhibited. Activated protein C did not inhibit endocytosis of thrombin-thrombomodulin complexes. Protein C inhibited both internalization and degradation of 125 I-thrombin and diisopropylphosphoryl (DIP) 125 I-thrombin in human lung cancer cells (A549). These effects were observed at protein C concentrations found in human plasma. Protein S had no effect on the inhibition of endocytosis of thrombin-thrombomodulin complexes by protein C. We propose that protein C may regulate the rate of endocytosis of thrombin-thrombomodulin complexes in vivo and thereby control the capacity for endothelium to activate protein C

  17. A label-free and high sensitive aptamer biosensor based on hyperbranched polyester microspheres for thrombin detection

    International Nuclear Information System (INIS)

    Sun, Chong; Han, Qiaorong; Wang, Daoying; Xu, Weimin; Wang, Weijuan; Zhao, Wenbo; Zhou, Min

    2014-01-01

    Highlights: • A label-free thrombin aptamer biosensor applied in whole blood has been developed. • The aptamer biosensor showed a wide detection range and a low detection limit. • The antibiofouling idea utilized for biosensor is significant for diagnostics. - Abstract: In this paper, we have synthesized hyperbranched polyester microspheres with carboxylic acid functional groups (HBPE-CA) and developed a label-free electrochemical aptamer biosensor using thrombin-binding aptamer (TBA) as receptor for the measurement of thrombin in whole blood. The indium tin oxide (ITO) electrode surface modified with HBPE-CA microspheres was grafted with TBA, which has excellent binding affinity and selectivity for thrombin. Binding of the thrombin at the modified ITO electrode surface greatly restrained access of electrons for a redox probe of [Fe(CN) 6 ] 3−/4− . Moreover, the aptamer biosensor could be used for detection of thrombin in whole blood, a wide detection range (10 fM–100 nM) and a detection limit on the order of 0.90 fM were demonstrated. Control experiments were also carried out by using bull serum albumin (BSA) and lysozyme in the absence of thrombin. The good stability and repeatability of this aptamer biosensor were also proved. We expect that this demonstration will lead to the development of highly sensitive label-free sensors based on aptamer with lower cost than current technology. The integration of the technologies, which include anticoagulant, sensor and nanoscience, will bring significant input to high-performance biosensors relevant to diagnostics and therapy of interest for human health

  18. A label-free and high sensitive aptamer biosensor based on hyperbranched polyester microspheres for thrombin detection

    Energy Technology Data Exchange (ETDEWEB)

    Sun, Chong [Jiangsu Key Laboratory of Biofunctional Materials, Biomedical Functional Materials Collaborative Innovation Center, College of Chemistry and Materials Science, Nanjing Normal University, Nanjing 210023 (China); Institute of Agricultural Products Processing, Jiangsu Academy of Agricultural Sciences, Nanjing 210014 (China); Han, Qiaorong [Jiangsu Key Laboratory of Biofunctional Materials, Biomedical Functional Materials Collaborative Innovation Center, College of Chemistry and Materials Science, Nanjing Normal University, Nanjing 210023 (China); Wang, Daoying; Xu, Weimin [Institute of Agricultural Products Processing, Jiangsu Academy of Agricultural Sciences, Nanjing 210014 (China); Wang, Weijuan [Jiangsu Key Laboratory of Biofunctional Materials, Biomedical Functional Materials Collaborative Innovation Center, College of Chemistry and Materials Science, Nanjing Normal University, Nanjing 210023 (China); Zhao, Wenbo, E-mail: zhaowenbo@njnu.edu.cn [Jiangsu Key Laboratory of Biofunctional Materials, Biomedical Functional Materials Collaborative Innovation Center, College of Chemistry and Materials Science, Nanjing Normal University, Nanjing 210023 (China); Zhou, Min, E-mail: zhouminnju@126.com [Department of Vascular Surgery, the Affiliated Drum Tower Hospital of Nanjing University Medical School, Nanjing 210008 (China)

    2014-11-19

    Highlights: • A label-free thrombin aptamer biosensor applied in whole blood has been developed. • The aptamer biosensor showed a wide detection range and a low detection limit. • The antibiofouling idea utilized for biosensor is significant for diagnostics. - Abstract: In this paper, we have synthesized hyperbranched polyester microspheres with carboxylic acid functional groups (HBPE-CA) and developed a label-free electrochemical aptamer biosensor using thrombin-binding aptamer (TBA) as receptor for the measurement of thrombin in whole blood. The indium tin oxide (ITO) electrode surface modified with HBPE-CA microspheres was grafted with TBA, which has excellent binding affinity and selectivity for thrombin. Binding of the thrombin at the modified ITO electrode surface greatly restrained access of electrons for a redox probe of [Fe(CN){sub 6}]{sup 3−/4−}. Moreover, the aptamer biosensor could be used for detection of thrombin in whole blood, a wide detection range (10 fM–100 nM) and a detection limit on the order of 0.90 fM were demonstrated. Control experiments were also carried out by using bull serum albumin (BSA) and lysozyme in the absence of thrombin. The good stability and repeatability of this aptamer biosensor were also proved. We expect that this demonstration will lead to the development of highly sensitive label-free sensors based on aptamer with lower cost than current technology. The integration of the technologies, which include anticoagulant, sensor and nanoscience, will bring significant input to high-performance biosensors relevant to diagnostics and therapy of interest for human health.

  19. Synthesis Polarimetry Calibration

    Science.gov (United States)

    Moellenbrock, George

    2017-10-01

    Synthesis instrumental polarization calibration fundamentals for both linear (ALMA) and circular (EVLA) feed bases are reviewed, with special attention to the calibration heuristics supported in CASA. Practical problems affecting modern instruments are also discussed.

  20. ORNL calibrations facility

    International Nuclear Information System (INIS)

    Berger, C.D.; Gupton, E.D.; Lane, B.H.; Miller, J.H.; Nichols, S.W.

    1982-08-01

    The ORNL Calibrations Facility is operated by the Instrumentation Group of the Industrial Safety and Applied Health Physics Division. Its primary purpose is to maintain radiation calibration standards for calibration of ORNL health physics instruments and personnel dosimeters. This report includes a discussion of the radioactive sources and ancillary equipment in use and a step-by-step procedure for calibration of those survey instruments and personnel dosimeters in routine use at ORNL

  1. Automating Finance

    Science.gov (United States)

    Moore, John

    2007-01-01

    In past years, higher education's financial management side has been riddled with manual processes and aging mainframe applications. This article discusses schools which had taken advantage of an array of technologies that automate billing, payment processing, and refund processing in the case of overpayment. The investments are well worth it:…

  2. Library Automation.

    Science.gov (United States)

    Husby, Ole

    1990-01-01

    The challenges and potential benefits of automating university libraries are reviewed, with special attention given to cooperative systems. Aspects discussed include database size, the role of the university computer center, storage modes, multi-institutional systems, resource sharing, cooperative system management, networking, and intelligent…

  3. Control console of the gamma calibration room

    International Nuclear Information System (INIS)

    Vilchis P, A.E.; Romero G, M.

    1999-01-01

    The Nuclear Centre of Mexico has a Ionizing Radiation Metrology Center (CMRI). This is in charge of the calibration in Mexico and Latin America of equipment dedicated to radiation measurement as industrial, medical as other fields. The importance to ensure that the equipment stay justly calibrated, it is imposed the necessity of automating the different rooms which the CMRI has. in this case it will be exposed the Calibration room for gamma radiation type. The operation of this application was carried out with the LabVIEW development platform and also in C language. The hardware associated is: personal computer with two cards using the 8255 device, 16 channels with optical isolation to manage input/output TTL type, 16 channels with optical isolation to management of charges to 127 V a.c., and 2 channels for 90V d.c. motors. (Author)

  4. Factor VIII S373L: mutation at P1' site confers thrombin cleavage resistance, causing mild haemophilia A.

    Science.gov (United States)

    Johnson, D J; Pemberton, S; Acquila, M; Mori, P G; Tuddenham, E G; O'Brien, D P

    1994-04-01

    A novel CRM+ mutation, factor VIII position 373 serine to leucine substitution (FVIII 373-Leu) was identified during a survey of Factor VIII (FVIII) mutations. We have purified the variant protein from the patient's plasma in order to allow further characterisation of the molecule. The CRM+ plasma contained 120% Factor VIII antigen (FVIII:Ag) and 6% Factor VIII coagulant activity (FVIII:C). After purification the mutant FVIII was subjected to thrombin proteolysis, and was thereby activated 5.6-fold compared with 7-fold for wild type molecule. Subsequently, spontaneous inactivation of the mutant was much slower than noted for wild type FVIII. Western blot analysis using monoclonal antibodies demonstrated that thrombin cleavage of FVIII 373-Leu at positions 740 and 1689 were normal but that cleavage at position 372 was completely absent. Crystallographic coordinates of the active site of thrombin complexed to fibrinopeptide A were used to explore possible mechanistic reasons for the failure of thrombin to cleave the mutant FVIII at position 372. Steric hindrance between the mutant side chain and the side chain of the P1 residue was apparent. We conclude that the functional defect of FVIII 373-Leu results from the inability of thrombin to cleave the mutant at position 372-373, and propose that this is due to steric hindrance by the side chain of leucine 373, preventing correct formation of the enzyme substrate complex.

  5. Fibrinogen and thrombin concentrations are critical for fibrin glue adherence in rat high-risk colon anastomoses

    Directory of Open Access Journals (Sweden)

    Eliseo Portilla-de Buen

    2014-04-01

    Full Text Available OBJECTIVE: Fibrin glues have not been consistently successful in preventing the dehiscence of high-risk colonic anastomoses. Fibrinogen and thrombin concentrations in glues determine their ability to function as sealants, healers, and/or adhesives. The objective of the current study was to compare the effects of different concentrations of fibrinogen and thrombin on bursting pressure, leaks, dehiscence, and morphology of high-risk ischemic colonic anastomoses using fibrin glue in rats. METHODS: Colonic anastomoses in adult female Sprague-Dawley rats (weight, 250-350 g treated with fibrin glue containing different concentrations of fibrinogen and thrombin were evaluated at post-operative day 5. The interventions were low-risk (normal or high-risk (ischemic end-to-end colonic anastomoses using polypropylene sutures and topical application of fibrinogen at high (120 mg/mL or low (40 mg/mL concentrations and thrombin at high (1000 IU/mL or low (500 IU/mL concentrations. RESULTS: Ischemia alone, anastomosis alone, or both together reduced the bursting pressure. Glues containing a low fibrinogen concentration improved this parameter in all cases. High thrombin in combination with low fibrinogen also improved adherence exclusively in low-risk anastomoses. No differences were detected with respect to macroscopic parameters, histopathology, or hydroxyproline content at 5 days post-anastomosis. CONCLUSIONS: Fibrin glue with a low fibrinogen content normalizes the bursting pressure of high-risk ischemic left-colon anastomoses in rats at day 5 after surgery.

  6. Key role of integrin α(IIb)β (3) signaling to Syk kinase in tissue factor-induced thrombin generation.

    Science.gov (United States)

    van der Meijden, Paola E J; Feijge, Marion A H; Swieringa, Frauke; Gilio, Karen; Nergiz-Unal, Reyhan; Hamulyák, Karly; Heemskerk, Johan W M

    2012-10-01

    The fibrin(ogen) receptor, integrin α(IIb)β(3), has a well-established role in platelet spreading, aggregation and clot retraction. How α(IIb)β(3) contributes to platelet-dependent coagulation is less well resolved. Here, we demonstrate that the potent suppressing effect of clinically used α(IIb)β(3) blockers on tissue factor-induced thrombin generation is linked to diminished platelet Ca(2+) responses and phosphatidylserine (PS) exposure. The same blockers suppress these responses in platelets stimulated with collagen and thrombin receptor agonists, whereas added fibrinogen potentiates these responses. In platelets spreading on fibrinogen, outside-in α(IIb)β(3) signaling similarly enhances thrombin-induced Ca(2+) rises and PS exposure. These responses are reduced in α(IIb)β(3)-deficient platelets from patients with Glanzmann's thrombasthenia. Furthermore, the contribution of α(IIb)β(3) to tissue factor-induced platelet Ca(2+) rises, PS exposure and thrombin generation in plasma are fully dependent on Syk kinase activity. Tyrosine phosphorylation analysis confirms a key role of Syk activation, which is largely but not exclusively dependent on α(IIb)β(3) activation. It is concluded that the majority of tissue factor-induced procoagulant activity of platelets relies on Syk activation and ensuing Ca(2+) signal generation, and furthermore that a considerable part of Syk activation relies on α(IIb)β(3) signaling. These results hence point to a novel role of Syk in integrin-dependent thrombin generation.

  7. Detection of Thrombin Based on Fluorescence Energy Transfer between Semiconducting Polymer Dots and BHQ-Labelled Aptamers

    Directory of Open Access Journals (Sweden)

    Yizhang Liu

    2018-02-01

    Full Text Available Carboxyl-functionalized semiconducting polymer dots (Pdots were synthesized as an energy donor by the nanoprecipitation method. A black hole quenching dye (BHQ-labelled thrombin aptamers was used as the energy acceptor, and fluorescence resonance energy transfer between the aptamers and Pdots was used for fluorescence quenching of the Pdots. The addition of thrombin restored the fluorescence intensity. Under the optimized experimental conditions, the fluorescence of the system was restored to the maximum when the concentration of thrombin reached 130 nM, with a linear range of 0–50 nM (R2 = 0.990 and a detection limit of 0.33 nM. This sensor was less disturbed by impurities, showing good specificity and signal response to thrombin, with good application in actual samples. The detection of human serum showed good linearity in the range of 0–30 nM (R2 = 0.997, with a detection limit of 0.56 nM and a recovery rate of 96.2–104.1%, indicating that this fluorescence sensor can be used for the detection of thrombin content in human serum.

  8. Magnetic relaxation switch and colorimetric detection of thrombin using aptamer-functionalized gold-coated iron oxide nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Liang Guohai; Cai Shaoyu; Zhang Peng [Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai 200433 (China); Peng Youyuan [Department of Chemistry, Quanzhou Normal University, Quanzhou 362000 (China); Chen Hui; Zhang Song [Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai 200433 (China); Kong Jilie, E-mail: jlkong@fudan.edu.cn [Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai 200433 (China)

    2011-03-18

    We describe a sensitive biosensing system combining magnetic relaxation switch diagnosis and colorimetric detection of human {alpha}-thrombin, based on the aptamer-protein interaction induced aggregation of Fe{sub 3}O{sub 4}-Au nanoparticles. To demonstrate the concept, gold-coated iron oxide nanoparticle was synthesized by iterative reduction of HAuCl{sub 4} onto the dextran-coated Fe{sub 3}O{sub 4} nanoparticles. The resulting core-shell structure had a flowerlike shape with pretty narrow size distribution (referred to as 'nanorose'). The two aptamers corresponding to human {alpha}-thrombin were conjugated separately to two distinct nanorose populations. Once a solution containing human {alpha}-thrombin was introduced, the nanoroses switched from a well dispersed state to an aggregated one, leading to a change in the spin-spin relaxation time (T{sub 2}) as well as the UV-Vis absorption spectra of the solution. Thus the qualitative and quantitative detection method for human {alpha}-thrombin was established. The dual-mode detection is clearly advantageous in obtaining a more reliable result; the detection range is widened as well. By using the dual-mode detection method, a detectable T{sub 2} change is observed with 1.0 nM human {alpha}-thrombin, and the detection range is from 1.6 nM to 30.4 nM.

  9. Preparation of chondroitin sulfate libraries containing disulfated disaccharide units and inhibition of thrombin by these chondroitin sulfates.

    Science.gov (United States)

    Numakura, Mario; Kusakabe, Noriko; Ishige, Kazuya; Ohtake-Niimi, Shiori; Habuchi, Hiroko; Habuchi, Osami

    2010-07-01

    Chondroitin sulfate (CS) containing GlcA-GalNAc(4,6-SO(4)) (E unit) and CS containing GlcA(2SO(4))-GalNAc(6SO(4)) (D unit) have been implicated in various physiological functions. However, it has been poorly understood how the structure and contents of disulfated disaccharide units in CS contribute to these functions. We prepared CS libraries containing E unit or D unit in various proportions by in vitro enzymatic reactions using recombinant GalNAc 4-sulfate 6-O-sulfotransferase and uronosyl 2-O-sulfotransferase, and examined their inhibitory activity toward thrombin. The in vitro sulfated CSs containing disulfated disaccharide units showed concentration-dependent direct inhibition of thrombin when the proportion of E unit or D unit in the CSs was above 15-17%. The CSs containing both E unit and D unit exhibited higher inhibitory activity toward thrombin than the CSs containing either E unit or D unit alone, if the proportion of the total disulfated disaccharide units of these CSs was comparable. The thrombin-catalyzed degradation of fibrinogen, a physiological substrate for thrombin, was also inhibited by the CS containing both E unit and D unit. These observations indicate that the enzymatically prepared CS libraries containing various amounts of disulfated disaccharide units appear to be useful for elucidating the physiological function of disulfated disaccharide units in CS.

  10. In vitro study of the role of thrombin in platelet rich plasma (PRP) preparation: utility for gel formation and impact in growth factors release.

    Science.gov (United States)

    Huber, Stephany Cares; Cunha Júnior, José Luiz Rosenberis; Montalvão, Silmara; da Silva, Letícia Queiroz; Paffaro, Aline Urban; da Silva, Francesca Aparecida Ramos; Rodrigues, Bruno Lima; Lana, José Fabio Santos Duarte; Annichino-Bizzacchi, Joyce Maria

    2016-01-01

    The use of PRP has been studied for different fields, with promising results in regenerative medicine. Until now, there is no study in the literature evaluating thrombin levels in serum, used as autologous thrombin preparation. Therefore, in the present study we evaluated the role played by different thrombin concentrations in PRP and the impact in the release of growth factors. Also, different activators for PRP gel formation were evaluated. Thrombin levels were measured in different autologous preparations: serum, L-PRP (PRP rich in leukocytes) and T-PRP (thrombin produced through PRP added calcium gluconate). L-PRP was prepared according to the literature, with platelets and leukocytes being quantified. The effect of autologous thrombin associated or not with calcium in PRP gel was determined by measuring the time of gel formation. The relationship between thrombin concentration and release of growth factors was determined by growth factors (PDGF-AA, VEGF and EGF) multiplex analysis. A similar concentration of thrombin was observed in serum, L-PRP and T-PRP (8.13 nM, 8.63 nM and 7.56 nM, respectively) with a high variation between individuals (CV%: 35.07, 43 and 58.42, respectively). T-PRP and serum with calcium chloride showed similar results in time to promote gel formation. The increase of thrombin concentrations (2.66, 8 and 24 nM) did not promote an increase in growth factor release. The technique of using serum as a thrombin source proved to be the most efficient and reproducible for promoting PRP gel formation, with some advantages when compared to other activation methods, as this technique is easier and quicker with no need of consuming part of PRP. Noteworthy, PRP activation using different thrombin concentrations did not promote a higher release of growth factors, appearing not to be necessary when PRP is used as a suspension.

  11. Renal function and plasma dabigatran level measured at trough by diluted thrombin time assay

    Directory of Open Access Journals (Sweden)

    Marta E. Martinuzzo

    2017-02-01

    Full Text Available Dabigatran etexilate (direct thrombin inhibitor is effective in preventing embolic stroke in patients with atrial fibrillation. It does not require laboratory control, but given the high renal elimination, its measurement in plasma is important in renal failure. The objectives of the study were to verify the analytical quality of the diluted thrombin time assay for measurement of dabigatran plasma concentration (cc, correlate cc with classic coagulation assays, prothrombin time (PT and activated partial thromboplastin time (APTT, and evaluate them according to the creatinine clearance (CLCr. Forty plasma samples of patients (34 consecutive and 6 suspected of drug accumulation receiving dabigatran at 150 (n = 19 or 110 (n = 21 mg/12 hours were collected. Blood samples were drawn at 10-14 hours of the last intake. Dabigatran concentration was determined by diluted thrombin time (HemosIl DTI, Instrumentation Laboratory (IL. PT and APTT (IL were performed on two fotooptical coagulometers, ACL TOP 300 and 500 (IL. DTI presented intra-assay coefficient of variation < 5.4% and inter-assay < 6%, linearity range 0-493 ng/ml. Patients' cc: median 83 (4-945 ng/ml. Individuals with CLCr in the lowest tertile (22.6-46.1 ml/min showed significantly higher median cc: 308 (49-945, compared to the average 72 (12-190 and highest tertile, 60 (4-118 ng/ml. Correlation between cc and APTT or PT were moderate, r2 = 0.59 and -0.66, p < 0.0001, respectively. DTI test allowed us to quantify plasma dabigatran levels, both in patients with normal or altered renal function, representing a useful tool in clinical situations such as renal failure, pre surgery or emergencies

  12. BjussuSP-I: a new thrombin-like enzyme isolated from Bothrops jararacussu snake venom.

    Science.gov (United States)

    Sant' Ana, Carolina D; Ticli, Fabio K; Oliveira, Leandro L; Giglio, Jose R; Rechia, Carem G V; Fuly, André L; Selistre de Araújo, Heloisa S; Franco, João J; Stabeli, Rodrigo G; Soares, Andreimar M; Sampaio, Suely V

    2008-11-01

    A thrombin-like enzyme named BjussuSP-I, isolated from B. jararacussu snake venom, is an acidic single chain glycoprotein with approximately 6% sugar, Mr=61,000 under reducing conditions and pI approximately 3.8, representing 1.09% of the chromatographic A(280) recovery. BjussuSP-I is a glycosylated serine protease containing both N-linked carbohydrates and sialic acid in its structure. BjussuSP-I showed a high clotting activity upon human plasma, which was inhibited by PMSF, leupeptin, heparin and 1,10-phenantroline. This enzyme showed high stability regarding coagulant activity when analyzed at different temperatures (-70 to 37 degrees C), pHs (4.5 to 8.0), and presence of two divalent metal ions (Ca(2+) and Mg(2+)). It also displayed TAME esterase and proteolytic activities toward natural (fibrinogen and fibrin) and synthetic (BAPNA) substrates, respectively, being also inhibited by PMSF and leupeptin. BjussuSP-I can induce production of polyclonal antibodies able to inhibit its clotting activity, but unable to inhibit its proteolytic activity on fibrinogen. The enzyme also showed crossed immunoreactivity against 11 venom samples of Bothrops, 1 of Crotalus, and 1 of Calloselasma snakes, in addition of LAAO isolated from B. moojeni venom. It displayed neither hemorrhagic, myotoxic, edema-inducing profiles nor proteolytic activity on casein. BjussuSP-I showed an N-terminal sequence (VLGGDECDINEHPFLA FLYS) similar to other thrombin-like enzymes from snake venoms. Based on its biochemical, enzymatic and pharmacological characteristics, BjussuSP-I was identified as a new thrombin-like enzyme isoform from Bothrops jararacussu snake venom.

  13. Enzyme-guided plasmonic biosensor based on dual-functional nanohybrid for sensitive detection of thrombin.

    Science.gov (United States)

    Yan, Jing; Wang, Lida; Tang, Longhua; Lin, Lei; Liu, Yang; Li, Jinghong

    2015-08-15

    Rapid and sensitive methodologies for the detection of protein are in urgent requirement for clinic diagnostics. Localized surface plasmon resonance (LSPR) of metal nanostructures has the potential to circumvent this problem due to its sensitive optical properties and strong electromagnetic near-field enhancements. In this work, an enzyme mediated plasmonic biosensor on the basis of a dual-functional nanohybrid was developed for the detection of thrombin. By utilizing LSPR-responsive nanohybrid and anaptamer-enzyme conjugated reporting probe, the sensing platform brings enhanced signal, stability as well as simplicity. Enzymatic reaction catalyzed the reduction of Au(3+) to Au° in situ, further leading to the rapid crystal growth of gold nanoparticles (AuNPs). The LSPR absorbance band and color changed company with the nanoparticle generation, which can be real-time monitoring by UV-visible spectrophotometer and naked eye. Nanohybrid constructed by gold and magnetic nanoparticles acts as a dual functional plasmonic unit, which not only plays the role of signal production, but also endows the sensor with the function of magnetic separation. Simultaneously, the introduction of enzyme effectively regulates the programming crystal growth of AuNPs. In addition, enzyme also serves as signal amplifier owing to its high catalysis efficiency. The response of the plasmonic sensor varies linearly with the logarithmic thrombin concentration up to 10nM with a limit of detection of 200 pM. The as-proposed strategy shows good analytical performance for thrombin determination. This simple, disposable method is promising in developing universal platforms for protein monitoring, drug discovery and point-of-care diagnostics. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. Minimally Invasive Therapy of Pseudoaneurysms of the Trunk: Application of Thrombin

    International Nuclear Information System (INIS)

    Schellhammer, Frank; Steinhaus, Daniele; Cohnen, Mathias; Hoppe, Jonas; Moedder, Ulrich; Fuerst, Guenter

    2008-01-01

    Thrombin injection has been proven to be successful in postcatheterization pseudoaneurysms. However, there are only a few reports on the treatment of pseudoaneurysms of the trunk. We report our first experiences using a percutaneous as well as an endovascular access. Eight iatrogenic pseudoaneurysms of the trunk (aorta, n = 4; pulmonary artery, n = 1; gastroduodenal artery, n = 1; left gastric artery, n = 1, renal artery, n = 1) were treated either percutaneously using CT guidance (n = 3) or via an endovascular access (n = 5). Noninvasive control angiograms were performed at day 1 and weeks 1 and 3 by either CT or MR angiography. The total volume of the pseudoaneurysms was 31.2 ± 23.1 ml on average, with a mean volume of the perfused aneurysmal lumen of 12.9 ± 7.2 ml. The maximum diameter was 4.1 ± 1.39 cm on average. In each case, the aneurysmal neck was not wider than 2 mm. One pseudoaneurysm occluded spontaneously following selective catheterization. The remaining pseudoaneurysms were successfully treated by injection of 765 ± 438.1 IU thrombin. In one individual, a nontarget embolization occurred, as well as an intervention-associated rupture of a pseudoaneurysm. High-grade stenoses of the donor artery were found in a different case. Only once was the endoluminal access converted to a percutaneous one. Thrombin injection might be a future first-line treatment of vascular lesions such as pseudoaneurysms of the trunk. In our experience both percutanous and endoluminal access are technically feasible and safe. However, further experiences are mandatory, especially concerning the question of dosage and long-term results

  15. Minimally invasive therapy of pseudoaneurysms of the trunk: application of thrombin.

    Science.gov (United States)

    Schellhammer, Frank; Steinhaus, Daniele; Cohnen, Mathias; Hoppe, Jonas; Mödder, Ulrich; Fürst, Günter

    2008-01-01

    Thrombin injection has been proven to be successful in postcatheterization pseudoaneurysms. However, there are only a few reports on the treatment of pseudoaneurysms of the trunk. We report our first experiences using a percutaneous as well as an endovascular access. Eight iatrogenic pseudoaneurysms of the trunk (aorta, n = 4; pulmonary artery, n = 1; gastroduodenal artery, n = 1; left gastric artery, n = 1, renal artery, n = 1) were treated either percutaneously using CT guidance (n = 3) or via an endovascular access (n = 5). Noninvasive control angiograms were performed at day 1 and weeks 1 and 3 by either CT or MR angiography. The total volume of the pseudoaneurysms was 31.2 +/- 23.1 ml on average, with a mean volume of the perfused aneurysmal lumen of 12.9 +/- 7.2 ml. The maximum diameter was 4.1 +/- 1.39 cm on average. In each case, the aneurysmal neck was not wider than 2 mm. One pseudoaneurysm occluded spontaneously following selective catheterization. The remaining pseudoaneurysms were successfully treated by injection of 765 +/- 438.1 IU thrombin. In one individual, a nontarget embolization occurred, as well as an intervention-associated rupture of a pseudoaneurysm. High-grade stenoses of the donor artery were found in a different case. Only once was the endoluminal access converted to a percutaneous one. Thrombin injection might be a future first-line treatment of vascular lesions such as pseudoaneurysms of the trunk. In our experience both percutanous and endoluminal access are technically feasible and safe. However, further experiences are mandatory, especially concerning the question of dosage and long-term results.

  16. Rapid Detection of Thrombin and Other Protease Activity Directly in Whole Blood

    Science.gov (United States)

    Yu, Johnson Chung Sing

    Thrombin is a serine protease that plays a key role in the clotting cascade to promote hemostasis following injury to the endothelium. From a clinical diagnostic perspective, in-vivo thrombin activity is linked to various blood clotting disorders, as well as cardiovascular disease (DVT, arteriosclerosis, etc). Thus, the ability to rapidly measure protease activity directly in whole blood will provide important new diagnostics, and clinical researchers with a powerful tool to further elucidate the relationship between circulating protease levels and disease. The ultimate goal is to design novel point of care (POC) diagnostic devices that are capable of monitoring protease activities directly in whole blood and biological sample. A charge-changing substrate specific to the thrombin enzyme was engineered and its functionality was confirmed by a series of experiments. This led to the preliminary design, construction, and testing of two device platforms deemed fully functional for the electrophoretic separation and focusing of charged peptide fragments. The concept of using the existing charge-changing substrate platform for bacterial protease detection was also investigated. Certain strains of E coli are associated with severe symptoms such as abdominal cramps, bloody diarrhea, and vomiting. The OmpT protease is expressed on the outer membrane of E coli and plays a role in the cleavage of antimicrobial peptides, the degradation of recombinant heterologous proteins, and the activation of plasminogen in the host. Thus, a synthetic peptide substrate specific to the OmpT protease was designed and modeled for the purpose of detecting E coli in biological sample.

  17. Comparing thrombin generation in patients with hemophilia A and patients on vitamin K antagonists.

    Science.gov (United States)

    de Koning, M L Y; Fischer, K; de Laat, B; Huisman, A; Ninivaggi, M; Schutgens, R E G

    2017-05-01

    Essentials It is unknown if hemophilia patients with atrial fibrillation need anticoagulation. Endogenous thrombin potentials (ETP) in hemophilia patients and patients on coumarins were compared. Severe hemophilia patients had comparable ETP to therapeutic international normalized ratio (INR). In non-severe hemophilia, 33% had higher ETP than therapeutic INR and may need anticoagulation. Click to hear Dr Negrier's perspective on global assays for assessing coagulation SUMMARY: Background It is unknown whether patients with hemophilia A with atrial fibrillation require treatment with vitamin K antagonists (VKAs) to the same extent as the normal population. Objective To compare hemostatic potential in hemophilia patients and patients on VKAs using thrombin generation (TG). Methods In this cross-sectional study, TG, initiated with 1pM tissue factor, was measured in 133 patients with severe (FVIII hemophilia A, 97 patients on a VKA with an international normalized ratio (INR) ≥ 1.5 and healthy controls. Endogenous thrombin potential (ETP) (nm*min) was compared according to FVIII level (hemophilia patients and patients on VKAs had lower median ETPs at 304 (196-449) and 176 (100-250), respectively. ETP was quite similar in severe hemophilia patients (185 [116-307]) and patients with a therapeutic INR (156 [90-225]). Compared with patients with therapeutic INR, ETP in patients with FVIII 1-19% and patients with FVIII 20-50% was higher at 296 (203-430) and 397 (219-632), respectively. All patients with therapeutic INR had an ETP hemophilia patients, 70% of patients with FVIII 1-19% and 52% of patients with FVIII 20-50% had an ETP hemophilia patients, TG was comparable to that in patients with a therapeutic INR. In one-third of non-severe hemophilia patients, TG was higher. These results suggest that anticoagulation therapy should be considered in a substantial proportion of non-severe hemophilia patients. © 2017 International Society on Thrombosis and Haemostasis.

  18. Differential proteolytic activation of factor VIII-von Willebrand factor complex by thrombin

    International Nuclear Information System (INIS)

    Hill-Eubanks, D.C.; Parker, C.G.; Lollar, P.

    1989-01-01

    Blood coagulation factor VIII (fVIII) is a plasma protein that is decreased or absent in hemophilia A. It is isolated as a mixture of heterodimers that contain a variably sized heavy chain and a common light chain. Thrombin catalyzes the activation of fVIII in a reaction that is associated with cleavages in both types of chain. The authors isolated a serine protease from Bothrops jararacussu snake venom that catalyzes thrombin-like heavy-chain cleavage but not light-chain cleavage in porcine fVIII as judged by NaDodSO 4 /PAGE and N-terminal sequence analysis. Using a plasma-free assay of the ability of activated 125 I-fVIII to function as a cofactor in the activation of factor X by factor IXa, they found that fVIII is activated by the venom enzyme. The venom enzyme-activated fVIII was isolated in stable form by cation-exchange HPLC. von Willebrand factor inhibited venom enzyme-activated fVIII but not thrombin-activated fVIII. These results suggest that the binding of fVIII to von Willebrand factor depends on the presence of an intact light chain and that activated fVIII must dissociate from von Willebrand factor to exert its cofactor effect. Thus, proteolytic activation of fVIII-von Willebrand factor complex appears to be differentially regulated by light-chain cleavage to dissociate the complex and heavy-chain cleavage to activate the cofactor function

  19. First steps in the direction of synthetic, allosteric, direct inhibitors of thrombin and factor Xa.

    Science.gov (United States)

    Verghese, Jenson; Liang, Aiye; Sidhu, Preet Pal Singh; Hindle, Michael; Zhou, Qibing; Desai, Umesh R

    2009-08-01

    Designing non-saccharide functional mimics of heparin is a major challenge. In this work, a library of small, aromatic molecules based on the sulfated DHP scaffold was synthesized and screened against thrombin and factor Xa. The results reveal that (i) selected monomeric benzofuran derivatives inhibit the two enzymes, albeit weakly; (ii) the two enzymes recognize different structural features in the benzofurans studied suggesting significant selectivity of recognition; and (iii) the mechanism of inhibition is allosteric. The molecules represent the first allosteric small molecule inhibitors of the two enzymes.

  20. First Steps in the Direction of Synthetic, Allosteric, Direct Inhibitors of Thrombin and Factor Xa

    Science.gov (United States)

    Verghese, Jenson; Liang, Aiye; Sidhu, Preet Pal Singh; Hindle, Michael; Zhou, Qibing; Desai, Umesh R.

    2009-01-01

    Designing non-saccharide functional mimics of heparin is a major challenge. In this work, a library of small, aromatic molecules based on the sulfated DHP scaffold was synthesized and screened against thrombin and factor Xa. The results reveal that i) selected monomeric benzofuran derivatives inhibit the two enzymes, albeit weakly; ii) the two enzymes recognize different structural features in the benzofurans studied suggesting significant selectivity of recognition; and iii) the mechanism of inhibition is allosteric. The molecules represent the first allosteric small molecule inhibitors of the two enzymes. PMID:19540113

  1. A study on the sensitivity of photogrammetric camera calibration and stitching

    CSIR Research Space (South Africa)

    De Villiers, J

    2014-11-01

    Full Text Available This paper presents a detailed simulation study of an automated robotic photogrammetric camera calibration system. The system performance was tested for sensitivity with regard to noise in the robot movement, camera mounting and image processing...

  2. Protein kinase C is differentially regulated by thrombin, insulin, and epidermal growth factor in human mammary tumor cells

    Energy Technology Data Exchange (ETDEWEB)

    Gomez, M.L.; Tellez-Inon, M.T. (Instituto de Ingenieria Genetica y Biologia Molecular, Buenos Aires (Argentina)); Medrano, E.E.; Cafferatta, E.G.A. (Instituto de Investigaciones Bioquimicas Fundacion Campomar, Buenos Aires (Argentina))

    1988-03-01

    The exposure of serum-deprived mammary tumor cells MCF-7 and T-47D to insulin, thrombin, and epidermal growth factor (EGF) resulted in dramatic modifications in the activity and in the translocation capacity of protein kinase C from cytosol to membrane fractions. Insulin induces a 600% activation of the enzyme after 5 h of exposure to the hormone in MCF-7 cells; thrombin either activates (200% in MCF-7) or down-regulates (in T-47D), and EGF exerts only a moderate effect. Thus, the growth factors studied modulate differentially the protein kinase C activity in human mammary tumor cells. The physiological significance of the results obtained are discussed in terms of the growth response elicited by insulin, thrombin, and EGF.

  3. Thermodynamic and biological evaluation of a thrombin binding aptamer modified with several unlocked nucleic acid (UNA) monomers and a 2′-C-piperazino-UNA monomer

    DEFF Research Database (Denmark)

    Jensen, Troels B.; Henriksen, Jonas Rosager; Rasmussen, Bjarne E.

    2011-01-01

    Thrombin binding aptamer is a DNA 15-mer which forms a G-quadruplex structure and possess promising anticoagulant properties due to specific interactions with thrombin. Herein we present the influence of a single 2′-C-piperazino-UNA residue and UNA residues incorporated in several positions on th...

  4. Incidence of thromboembolic events after use of gelatin-thrombin-based hemostatic matrix during intracranial tumor surgery.

    Science.gov (United States)

    Gazzeri, Roberto; Galarza, Marcelo; Conti, Carlo; De Bonis, Costanzo

    2018-01-01

    Association between the use of hemostatic agents made from collagen/gelatin mixed with thrombin and thromboembolic events in patients undergoing tumor resection has been suggested. This study evaluates the relationship between flowable hemostatic matrix and deep vein thrombosis in a large cohort of patients treated for brain tumor removal. The authors conducted a retrospective, multicenter, clinical review of all craniotomies for tumor removal performed between 2013 and 2014. Patients were classified in three groups: group I (flowable gelatin hemostatic matrix with thrombin), group II (gelatin hemostatic without thrombin), and group III (classical hemostatic). A total of 932 patients were selected: tumor pathology included 441 gliomas, 296 meningiomas, and 195 metastases. Thromboembolic events were identified in 4.7% of patients in which gelatin matrix with thrombin was applied, in 8.4% of patients with gelatin matrix without thrombin, and in 3.6% of cases with classical methods of hemostasis. Patients with venous thromboembolism had an increased proportion of high-grade gliomas (7.2%). Patients receiving a greater dose than 10 ml gelatin hemostatic had a higher rate of thromboembolic events. Intracranial hematoma requiring reintervention occurred in 19 cases: 4.5% of cases of group III, while reoperation was performed in 1.3 and 1.6% of patients in which gelatin matrix with or without thrombin was applied. Gelatin matrix hemostat is an efficacious tool for neurosurgeons in cases of difficult intraoperative bleeding during cranial tumor surgery. This study may help to identify those patients at high risk for developing thromboembolism and to treat them accordingly.

  5. Inhibition of thrombin by functionalized C60 nanoparticles revealed via in vitro assays and in silico studies.

    Science.gov (United States)

    Liu, Yanyan; Fu, Jianjie; Pan, Wenxiao; Xue, Qiao; Liu, Xian; Zhang, Aiqian

    2018-01-01

    The studies on the human toxicity of nanoparticles (NPs) are far behind the rapid development of engineered functionalized NPs. Fullerene has been widely used as drug carrier skeleton due to its reported low risk. However, different from other kinds of NPs, fullerene-based NPs (C 60 NPs) have been found to have an anticoagulation effect, although the potential target is still unknown. In the study, both experimental and computational methods were adopted to gain mechanistic insight into the modulation of thrombin activity by nine kinds of C 60 NPs with diverse surface chemistry properties. In vitro enzyme activity assays showed that all tested surface-modified C 60 NPs exhibited thrombin inhibition ability. Kinetic studies coupled with competitive testing using 3 known inhibitors indicated that six of the C 60 NPs, of greater hydrophobicity and hydrogen bond (HB) donor acidity or acceptor basicity, acted as competitive inhibitors of thrombin by directly interacting with the active site of thrombin. A simple quantitative nanostructure-activity relationship model relating the surface substituent properties to the inhibition potential was then established for the six competitive inhibitors. Molecular docking analysis revealed that the intermolecular HB interactions were important for the specific binding of C 60 NPs to the active site canyon, while the additional stability provided by the surface groups through van der Waals interaction also play a key role in the thrombin binding affinity of the NPs. Our results suggest that thrombin is a possible target of the surface-functionalized C 60 NPs relevant to their anticoagulation effect. Copyright © 2017. Published by Elsevier B.V.

  6. Deletion of the thrombin cleavage domain of osteopontin mediates breast cancer cell adhesion, proteolytic activity, tumorgenicity, and metastasis

    International Nuclear Information System (INIS)

    Beausoleil, Michel S; Schulze, Erika B; Goodale, David; Postenka, Carl O; Allan, Alison L

    2011-01-01

    Osteopontin (OPN) is a secreted phosphoprotein often overexpressed at high levels in the blood and primary tumors of breast cancer patients. OPN contains two integrin-binding sites and a thrombin cleavage domain located in close proximity to each other. To study the role of the thrombin cleavage site of OPN, MDA-MB-468 human breast cancer cells were stably transfected with either wildtype OPN (468-OPN), mutant OPN lacking the thrombin cleavage domain (468-ΔTC) or an empty vector (468-CON) and assessed for in vitro and in vivo functional differences in malignant/metastatic behavior. All three cell lines were found to equivalently express thrombin, tissue factor, CD44, αvβ5 integrin and β1 integrin. Relative to 468-OPN and 468-CON cells, 468-ΔTC cells expressing OPN with a deleted thrombin cleavage domain demonstrated decreased cell adhesion (p < 0.001), decreased mRNA expression of MCAM, maspin and TRAIL (p < 0.01), and increased uPA expression and activity (p < 0.01) in vitro. Furthermore, injection of 468-ΔTC cells into the mammary fat pad of nude mice resulted in decreased primary tumor latency time (p < 0.01) and increased primary tumor growth and lymph node metastatic burden (p < 0.001) compared to 468-OPN and 468-CON cells. The results presented here suggest that expression of thrombin-uncleavable OPN imparts an early tumor formation advantage as well as a metastatic advantage for breast cancer cells, possibly due to increased proteolytic activity and decreased adhesion and apoptosis. Clarification of the mechanisms responsible for these observations and the translation of this knowledge into the clinic could ultimately provide new therapeutic opportunities for combating breast cancer

  7. Shc adaptor proteins are key transducers of mitogenic signaling mediated by the G protein-coupled thrombin receptor

    DEFF Research Database (Denmark)

    Chen, Y; Grall, D; Salcini, A E

    1996-01-01

    The serine protease thrombin activates G protein signaling systems that lead to Ras activation and, in certain cells, proliferation. Whereas the steps leading to Ras activation by G protein-coupled receptors are not well defined, the mechanisms of Ras activation by receptor tyrosine kinases have......-insensitive proteins appear to mediate this effect, since (i) pertussis toxin pre-treatment of cells does not blunt the action of thrombin and (ii) Shc phosphorylation on tyrosine can be stimulated by the muscarinic m1 receptor. Shc phosphorylation does not appear to involve protein kinase C, since the addition of 4...

  8. Calibration of Flick standards

    International Nuclear Information System (INIS)

    Thalmann, Ruedi; Spiller, Jürg; Küng, Alain; Jusko, Otto

    2012-01-01

    Flick standards or magnification standards are widely used for an efficient and functional calibration of the sensitivity of form measuring instruments. The results of a recent measurement comparison have shown to be partially unsatisfactory and revealed problems related to the calibration of these standards. In this paper the influence factors for the calibration of Flick standards using roundness measurement instruments are discussed in detail, in particular the bandwidth of the measurement chain, residual form errors of the device under test, profile distortions due to the diameter of the probing element and questions related to the definition of the measurand. The different contributions are estimated using simulations and are experimentally verified. Also alternative methods to calibrate Flick standards are investigated. Finally the practical limitations of Flick standard calibration are shown and the usability of Flick standards both to calibrate the sensitivity of roundness instruments and to check the filter function of such instruments is analysed. (paper)

  9. Computerized 50 liter volume calibration system

    International Nuclear Information System (INIS)

    Proffitt, T.H.

    1990-01-01

    A system has been designed for the Savannah River Site that will be used to calibrate product shipping containers. For accountability purposes, it is necessary that these containers be calibrated to a very high precision. The Computerized 50 Liter Volume Calibration System (CVCS), which is based on the Ideal Gas Law (IGL), will use reference volumes with precision of no less ±0.03%, and helium to calibrate the containers to have a total error of no greater than ±0.10%. A statistical interpretation of the system has given a theoretical total calculated error of ±0.08%. Tests with the system will be performed once fabrication is complete to experimentally verify the calculated error. Since the total error was calculated using the worst case scenario, the actual error should be significantly less than the calculated value. The computer controlled, totally automated system is traceable to the National Institute of Standards and Technology. The design, calibration procedure, and statistical interpretation of the system will be discussed. 1 ref

  10. Calibration of areal surface topography measuring instruments

    Science.gov (United States)

    Seewig, J.; Eifler, M.

    2017-06-01

    The ISO standards which are related to the calibration of areal surface topography measuring instruments are the ISO 25178-6xx series which defines the relevant metrological characteristics for the calibration of different measuring principles and the ISO 25178-7xx series which defines the actual calibration procedures. As the field of areal measurement is however not yet fully standardized, there are still open questions to be addressed which are subject to current research. Based on this, selected research results of the authors in this area are presented. This includes the design and fabrication of areal material measures. For this topic, two examples are presented with the direct laser writing of a stepless material measure for the calibration of the height axis which is based on the Abbott- Curve and the manufacturing of a Siemens star for the determination of the lateral resolution limit. Based on these results, as well a new definition for the resolution criterion, the small scale fidelity, which is still under discussion, is presented. Additionally, a software solution for automated calibration procedures is outlined.

  11. In vitro effects of recombinant activated factor VII on thrombin generation and coagulation following inhibition of platelet procoagulant activity by prasugrel.

    Science.gov (United States)

    Mazzeffi, Michael; Szlam, Fania; Jakubowski, Joseph A; Tanaka, Kenichi A; Sugidachi, Atsuhiro; Levy, Jerrold H

    2013-07-01

    Prasugrel is a thienopyridyl P2Y12 antagonist with potent antiplatelet effects. At present, little is known about its effects on thrombin generation or what strategies may emergently reverse its anticoagulant effects. In the current study we evaluated whether recombinant activated factor VII may reverse prasugrel induced effects and increase thrombin generation in an in vitro model. The effect of prasugrel active metabolite, PAM (R-138727), was evaluated on platelet aggregation, thrombin generation, and rotational thromboelastometry parameters using blood from 20 healthy volunteers. Additionally, we evaluated the effects of adenosine diphosphate (ADP) and recombinant activated factor VII on restoring these parameters towards baseline values. PAM reduced maximum platelet aggregation and led to platelet disaggregation. It also decreased peak thrombin, increased lag time, and increased time to peak thrombin. Treatment with recombinant activated factor VII restored all three parameters of thrombin generation towards baseline. ADP decreased lag time and time to peak thrombin, but had no effect on peak thrombin. When recombinant activated factor VII and ADP were combined they had a greater effect on thrombin parameters than either drug alone. PAM also increased thromboelastometric clotting time and clot formation time, but had no effect on maximum clot firmness. Treatment with either recombinant activated factor VII or ADP restored these values towards baseline. Recombinant activated factor VII restores thrombin generation in the presence of PAM. In patients taking prasugrel with life-threatening refractory bleeding it has the potential to be a useful therapeutic approach. Additional clinical studies are needed to validate our findings. Copyright © 2013 Elsevier Ltd. All rights reserved.

  12. RF impedance measurement calibration

    International Nuclear Information System (INIS)

    Matthews, P.J.; Song, J.J.

    1993-01-01

    The intent of this note is not to explain all of the available calibration methods in detail. Instead, we will focus on the calibration methods of interest for RF impedance coupling measurements and attempt to explain: (1). The standards and measurements necessary for the various calibration techniques. (2). The advantages and disadvantages of each technique. (3). The mathematical manipulations that need to be applied to the measured standards and devices. (4). An outline of the steps needed for writing a calibration routine that operated from a remote computer. For further details of the various techniques presented in this note, the reader should consult the references

  13. FVIIa-sTF and Thrombin Inhibitory Activities of Compounds Isolated from Microcystis aeruginosa K-139

    Directory of Open Access Journals (Sweden)

    Andrea Roxanne J. Anas

    2017-08-01

    Full Text Available The rise of bleeding and bleeding complications caused by oral anticoagulant use are serious problems nowadays. Strategies that block the initiation step in blood coagulation involving activated factor VII-tissue factor (fVIIa-TF have been considered. This study explores toxic Microcystis aeruginosa K-139, from Lake Kasumigaura, Ibaraki, Japan, as a promising cyanobacterium for isolation of fVIIa-sTF inhibitors. M. aeruginosa K-139 underwent reversed-phase solid-phase extraction (ODS-SPE from 20% MeOH to MeOH elution with 40%-MeOH increments, which afforded aeruginosin K-139 in the 60% MeOH fraction; micropeptin K-139 and microviridin B in the MeOH fraction. Aeruginosin K-139 displayed an fVIIa-sTF inhibitory activity of ~166 µM, within a 95% confidence interval. Micropeptin K-139 inhibited fVIIa-sTF with EC50 10.62 µM, which was more efficient than thrombin inhibition of EC50 26.94 µM. The thrombin/fVIIa-sTF ratio of 2.54 in micropeptin K-139 is higher than those in 4-amidinophenylmethane sulfonyl fluoride (APMSF and leupeptin, when used as positive controls. This study proves that M. aeruginosa K-139 is a new source of fVIIa-sTF inhibitors. It also opens a new avenue for micropeptin K-139 and related depsipeptides as fVIIa-sTF inhibitors.

  14. Computational study of some benzamidine-based inhibitors of thrombin-like snake venom proteinases

    Science.gov (United States)

    Henriques, Elsa S.; Nascimento, Marco A. C.; Ramos, Maria João

    Pit viper venoms contain a number of serine proteinases that, despite their observed coagulant thrombin-like action in vitro, exhibit a paradoxical benign defibrinogenating (anticoagulant) action in vivo, with clinical applications in preventing thrombi and improved blood circulation. Considering that several benzamidine-based inhibitors, some highly selective to thrombin, also inhibit the enzymatic activity of such venombins, the modeling of their enzyme-inhibitor interactions could provide valuable information on the topological factors that determine the divergences in activity. The first step, and the object of the present study, was to derive the necessary set of parameters, consistent with the CHARMM force field, and to perform molecular dynamics (MD) simulations on a few selected representatives of the inhibitors in question under physiological conditions. Bonding and van der Waals parameters were derived by analogy to similar ones in the existing force field. Net atomic charges were obtained with a restrained fitting to the molecular electrostatic potential generated at B3LYP/6-31G(d) level. The parameters were refined to reproduce the available experimental geometries and crystal data, and the MD simulations of the free inhibitors in aqueous solution at 298 K provided an insightful description of their available conformational space.

  15. Proteolytic signatures define unique thrombin-derived peptides present in human wound fluid in vivo.

    Science.gov (United States)

    Saravanan, Rathi; Adav, Sunil S; Choong, Yeu Khai; van der Plas, Mariena J A; Petrlova, Jitka; Kjellström, Sven; Sze, Siu Kwan; Schmidtchen, Artur

    2017-10-13

    The disease burden of failing skin repair and non-healing ulcers is extensive. There is an unmet need for new diagnostic approaches to better predict healing activity and wound infection. Uncontrolled and excessive protease activity, of endogenous or bacterial origin, has been described as a major contributor to wound healing impairments. Proteolytic peptide patterns could therefore correlate and "report" healing activity and infection. This work describes a proof of principle delineating a strategy by which peptides from a selected protein, human thrombin, are detected and attributed to proteolytic actions. With a particular focus on thrombin-derived C-terminal peptides (TCP), we show that distinct peptide patterns are generated in vitro by the human S1 peptidases human neutrophil elastase and cathepsin G, and the bacterial M4 peptidases Pseudomonas aeruginosa elastase and Staphylococcus aureus aureolysin, respectively. Corresponding peptide sequences were identified in wound fluids from acute and non-healing ulcers, and notably, one peptide, FYT21 (FYTHVFRLKKWIQKVIDQFGE), was only present in wound fluid from non-healing ulcers colonized by P. aeruginosa and S. aureus. Our result is a proof of principle pointing at the possibility of defining peptide biomarkers reporting distinct proteolytic activities, of potential implication for improved diagnosis of wound healing and infection.

  16. A sensitive HIV-1 envelope induced fusion assay identifies fusion enhancement of thrombin

    International Nuclear Information System (INIS)

    Cheng, De-Chun; Zhong, Guo-Cai; Su, Ju-Xiang; Liu, Yan-Hong; Li, Yan; Wang, Jia-Ye; Hattori, Toshio; Ling, Hong; Zhang, Feng-Min

    2010-01-01

    To evaluate the interaction between HIV-1 envelope glycoprotein (Env) and target cell receptors, various cell-cell-fusion assays have been developed. In the present study, we established a novel fusion system. In this system, the expression of the sensitive reporter gene, firefly luciferase (FL) gene, in the target cells was used to evaluate cell fusion event. Simultaneously, constitutively expressed Renilla luciferase (RL) gene was used to monitor effector cell number and viability. FL gave a wider dynamic range than other known reporters and the introduction of RL made the assay accurate and reproducible. This system is especially beneficial for investigation of potential entry-influencing agents, for its power of ruling out the false inhibition or enhancement caused by the artificial cell-number variation. As a case study, we applied this fusion system to observe the effect of a serine protease, thrombin, on HIV Env-mediated cell-cell fusion and have found the fusion enhancement activity of thrombin over two R5-tropic HIV strains.

  17. Thrombin-activatable fibrinolysis inhibitor is degraded by Salmonella enterica and Yersinia pestis.

    Science.gov (United States)

    Valls Serón, M; Haiko, J; DE Groot, P G; Korhonen, T K; Meijers, J C M

    2010-10-01

     Pathogenic bacteria modulate the host coagulation system to evade immune responses or to facilitate dissemination through extravascular tissues. In particular, the important bacterial pathogens Salmonella enterica and Yersinia pestis intervene with the plasminogen/fibrinolytic system. Thrombin-activatable fibrinolysis inhibitor (TAFI) has anti-fibrinolytic properties as the active enzyme (TAFIa) removes C-terminal lysine residues from fibrin, thereby attenuating accelerated plasmin formation.  Here, we demonstrate inactivation and cleavage of TAFI by homologous surface proteases, the omptins Pla of Y. pestis and PgtE of S. enterica. We show that omptin-expressing bacteria decrease TAFI activatability by thrombin-thrombomodulin and that the anti-fibrinolytic potential of TAFIa was reduced by recombinant Escherichia coli expressing Pla or PgtE. The functional impairment resulted from C-terminal cleavage of TAFI by the omptins.  Our results indicate that TAFI is degraded directly by the omptins PgtE of S. enterica and Pla of Y. pestis. This may contribute to the ability of PgtE and Pla to damage tissue barriers, such as fibrin, and thereby to enhance spread of S. enterica and Y. pestis during infection. © 2010 International Society on Thrombosis and Haemostasis.

  18. High throughput protease profiling comprehensively defines active site specificity for thrombin and ADAMTS13.

    Science.gov (United States)

    Kretz, Colin A; Tomberg, Kärt; Van Esbroeck, Alexander; Yee, Andrew; Ginsburg, David

    2018-02-12

    We have combined random 6 amino acid substrate phage display with high throughput sequencing to comprehensively define the active site specificity of the serine protease thrombin and the metalloprotease ADAMTS13. The substrate motif for thrombin was determined by >6,700 cleaved peptides, and was highly concordant with previous studies. In contrast, ADAMTS13 cleaved only 96 peptides (out of >10 7 sequences), with no apparent consensus motif. However, when the hexapeptide library was substituted into the P3-P3' interval of VWF73, an exosite-engaging substrate of ADAMTS13, 1670 unique peptides were cleaved. ADAMTS13 exhibited a general preference for aliphatic amino acids throughout the P3-P3' interval, except at P2 where Arg was tolerated. The cleaved peptides assembled into a motif dominated by P3 Leu, and bulky aliphatic residues at P1 and P1'. Overall, the P3-P2' amino acid sequence of von Willebrand Factor appears optimally evolved for ADAMTS13 recognition. These data confirm the critical role of exosite engagement for substrates to gain access to the active site of ADAMTS13, and define the substrate recognition motif for ADAMTS13. Combining substrate phage display with high throughput sequencing is a powerful approach for comprehensively defining the active site specificity of proteases.

  19. Plasma thrombin-cleaved osteopontin elevation after carotid artery stenting in symptomatic ischemic stroke patients

    International Nuclear Information System (INIS)

    Kurata, Mie; Okura, Takafumi; Kumon, Yoshiaki; Tagawa, Masahiko; Watanabe, Hideaki; Miyazaki, Tatsuhiko; Higaki, Jitsuo; Nose, Masato; Nakahara, Toshinori

    2012-01-01

    Atherothrombosis is the primary pathophysiology that underlies ischemic cerebral infarction. Osteopontin (OPN) is produced in atherosclerotic lesions and is cleaved by activated thrombin. We hypothesized that the rupture or damage of an unstable atherosclerotic plaque increases plasma levels of thrombin-cleaved OPN (trOPN). This study included 90 patients who received carotid angioplasty with stenting (CAS), 23 patients with essential hypertension (EHT) and 10 patients who were treated with carotid endarterectomy (CEA). The CAS patient group included 36 patients that had pre- and post-operative blood tests, diffusion-weighted imaging (DWI) using cerebral MRIs and estimated thrombus debris within the protection device. Immunohistochemistry of CEA specimens revealed that trOPN was detected around intra-plaque vessels. The highest tertile of plasma trOPN levels in CAS patients was higher than trOPN levels in EHT patients. Post-operative trOPN levels were significantly higher in symptomatic compared with asymptomatic patients (P=0.003). New ipsilateral DWI-positive patients revealed higher post-operative trOPN levels (P=0.003) and a higher grade of thrombi (P<0.001) than DWI-negative patients. TrOPN may be a novel biomarker that reflects the atherothrombotic status in ischemic stroke. (author)

  20. Correlation between Interleukin-6 and Thrombin-Antithrombin III Complex Levels in Retinal Diseases.

    Science.gov (United States)

    Ehrlich, Rita; Zahavi, Alon; Axer-Siegel, Ruth; Budnik, Ivan; Dreznik, Ayelet; Dahbash, Mor; Nisgav, Yael; Megiddo, Elinor; Kenet, Gili; Weinberger, Dov; Livnat, Tami

    2017-09-01

    This study aims to evaluate and correlate the levels of interleukin-6 (IL-6) and thrombin-antithrombin III complex (TAT) in the vitreous of patients with different vitreoretinal pathologies. Vitreous samples were collected from 78 patients scheduled for pars plana vitrectomy at a tertiary medical center. Patients were divided by the underlying vitreoretinal pathophysiology, as follows: macular hole (MH)/epiretinal membrane (ERM) (n = 26); rhegmatogenous retinal detachment (RRD) (n = 32); and proliferative diabetic retinopathy (PDR) (n = 20). Levels of IL-6 and TAT were measured by enzyme-linked immunosorbent assay and compared among the groups. A significant difference was found in the vitreal IL-6 and TAT levels between the MH/ERM group and both the PDR and RRD groups (P Diabetes was associated with higher IL-6 levels in the RRD group. Different relationships between the IL-6 and TAT levels were revealed in patients with different ocular pathologies. Our results imply that variations in vitreal TAT level may be attributable not only to an inflammatory reaction or blood-retinal barrier breakdown, but also to intraocular tissue-dependent regulation of thrombin.

  1. Infection-Induced Thrombin Production: A Potential Novel Mechanism for Preterm Premature Rupture of Membranes (PPROM).

    Science.gov (United States)

    Feng, Liping; Allen, Terrence K; Marinello, William P; Murtha, Amy P

    2018-04-13

    Preterm premature rupture of membranes (PPROM) is a leading contributor to maternal and neonatal morbidity and mortality. Epidemiologic and experimental studies have demonstrated that thrombin causes fetal membrane weakening and subsequently PPROM. Although blood is suspected as the likely source of thrombin in fetal membranes and amniotic fluid of patients with PPROM, this has not been proven. Ureaplasma Parvum (U. parvum) is emerging as a pathogen involved in prematurity, including PPROM, but until now, prothrombin production directly induced by bacteria in fetal membranes has not been described. This study was designed to investigate whether U. parvum exposure can induce prothrombin production in fetal membranes cells. Primary fetal membrane cells (amnion epithelial, chorion trophoblast, and decidua stromal) or full-thickness fetal membrane tissue explants from elective, term, uncomplicated cesarean deliveries were harvested. Cells or tissue explants were infected with live U. parvum (1 x 10 5 , 1 x 10 6 , or 1 x 10 7 colony forming units (cfu)/ml) or lipopolysaccharide (Escherichia coli J5, L-5014, Sigma, 100 ng/ml or 1000 ng/ml) for 24 hours. Tissue explants were fixed for immunohistochemistry staining of thrombin/prothrombin. Fetal membrane cells were fixed for confocal immunofluorescent staining of the biomarkers of fetal membrane cell types and thrombin/prothrombin. Protein and mRNA were harvested from the cells and tissue explants for Western blot or qRT-PCR to quantify thrombin/prothrombin protein or mRNA production, respectively. Data are presented as mean values ± standard errors of mean. Data were analyzed using one-way ANOVA with post hoc Dunnett's test. Prothrombin production and localization was confirmed by Western blot and immunostainings in all primary fetal membrane cells and tissue explants. Immunofluorescence observations revealed a perinuclear localization of prothrombin in amnion epithelial cells. Localization of prothrombin in chorion and

  2. Plant automation

    International Nuclear Information System (INIS)

    Christensen, L.J.; Sackett, J.I.; Dayal, Y.; Wagner, W.K.

    1989-01-01

    This paper describes work at EBR-II in the development and demonstration of new control equipment and methods and associated schemes for plant prognosis, diagnosis, and automation. The development work has attracted the interest of other national laboratories, universities, and commercial companies. New initiatives include use of new control strategies, expert systems, advanced diagnostics, and operator displays. The unique opportunity offered by EBR-II is as a test bed where a total integrated approach to automatic reactor control can be directly tested under real power plant conditions

  3. Inherited Anti-Thrombin Deficiency in A Malay-Malaysian Family: A Missense Mutation at Nucleotide g.13267C>A aka anti-thrombin Budapest 5 (p.Pro439Thr) of the SERPINC 1 gene.

    Science.gov (United States)

    Norlelawati, A T; Rusmawati, I; Naznin, M; Nur Nadia, O; Rizqan Aizzani, R; Noraziana, A W

    2014-02-01

    Inherited anti-thrombin deficiency is an autosomal dominant disorder which is associated with increased risk for venous thromboembolism (VTE). This condition is very rare in Malaysia and there has been no documented report. Thus, the aim of the present study is to investigate the type of an inherited anti-thrombin deficiency mutation in a 25-year-old Malay woman who presented with deep vein thrombosis in her first pregnancy. DNA was extracted from the patient's blood sample and buccal mucosal swabs from family members. Polymerase chain reaction(PCR) assays were designed to cover all seven exons of the serpin peptidase inhibitor, clade C (antithrombin), member 1 (SERPINC1) gene; and the products were subjected to DNA sequencing. Sequences were referred to NCBI Reference Sequence: NG_012462.1. A heterozygous substitution mutation at nucleotide position 13267 (CCT->ACT) was identified in the patient and two other family members, giving a possible change of codon 439 (Pro→Thr) also known as anti-thrombin Budapest 5. The genotype was absent in 90 healthy controls. The study revealed a heterozygous antithrombin Budapest 5 mutation in SERPINC 1 giving rise to a possible anti-thrombin deficiency in a Malay-Malaysian family.

  4. Reducing the Variance of Intrinsic Camera Calibration Results in the ROS Camera_Calibration Package

    Science.gov (United States)

    Chiou, Geoffrey Nelson

    The intrinsic calibration of a camera is the process in which the internal optical and geometric characteristics of the camera are determined. If accurate intrinsic parameters of a camera are known, the ray in 3D space that every point in the image lies on can be determined. Pairing with another camera allows for the position of the points in the image to be calculated by intersection of the rays. Accurate intrinsics also allow for the position and orientation of a camera relative to some world coordinate system to be calculated. These two reasons for having accurate intrinsic calibration for a camera are especially important in the field of industrial robotics where 3D cameras are frequently mounted on the ends of manipulators. In the ROS (Robot Operating System) ecosystem, the camera_calibration package is the default standard for intrinsic camera calibration. Several researchers from the Industrial Robotics & Automation division at Southwest Research Institute have noted that this package results in large variances in the intrinsic parameters of the camera when calibrating across multiple attempts. There are also open issues on this matter in their public repository that have not been addressed by the developers. In this thesis, we confirm that the camera_calibration package does indeed return different results across multiple attempts, test out several possible hypothesizes as to why, identify the reason, and provide simple solution to fix the cause of the issue.

  5. Calibration of moisture monitors

    International Nuclear Information System (INIS)

    Gutierrez, R.L.

    1979-02-01

    A method for calibrating an aluminum oxide hygrometer against an optical chilled mirror dew-point hygrometer has been established. A theoretical cross-point line of dew points from both hygrometers and a maximum moisture content of 10 ppM/sub v/ are used to define an area for calibrating the sensor probes of the aluminum oxide hygrometer

  6. Site Calibration report

    DEFF Research Database (Denmark)

    Yordanova, Ginka; Vesth, Allan

    The report describes site calibration measurements carried out on a site in Denmark. The measurements are carried out in accordance to Ref. [1]. The site calibration is carried out before a power performance measurement on a given turbine to clarify the influence from the terrain on the ratio...

  7. Topics in Statistical Calibration

    Science.gov (United States)

    2014-03-27

    Natural cubic spline speed di st 110 B.2 The calibrate function The most basic calibration problem, the one often encountered in more advanced ...0040-1706, 1537-2723. A. M. Mood, F. A. Graybill, and D. C. Boes. Introduction to the Theory of Statistics. McGraw-Hill, Auckland , U.A, 1974. ISBN

  8. Thrombin Time

    Science.gov (United States)

    ... information]. Available online at http://www2.massgeneral.org/pathology/coagbook/CO004600.htm through http://www2.massgeneral.org . Accessed July 2010. Dugdale, D. (Updated 2009 March 2). Congenital afibrinogenemia. MedlinePlus Medical Encyclopedia [On-line ...

  9. WIDAFELS flexible automation systems

    International Nuclear Information System (INIS)

    Shende, P.S.; Chander, K.P.; Ramadas, P.

    1990-01-01

    After discussing the various aspects of automation, some typical examples of various levels of automation are given. One of the examples is of automated production line for ceramic fuel pellets. (M.G.B.)

  10. An Automation Planning Primer.

    Science.gov (United States)

    Paynter, Marion

    1988-01-01

    This brief planning guide for library automation incorporates needs assessment and evaluation of options to meet those needs. A bibliography of materials on automation planning and software reviews, library software directories, and library automation journals is included. (CLB)

  11. Sandia WIPP calibration traceability

    Energy Technology Data Exchange (ETDEWEB)

    Schuhen, M.D. [Sandia National Labs., Albuquerque, NM (United States); Dean, T.A. [RE/SPEC, Inc., Albuquerque, NM (United States)

    1996-05-01

    This report summarizes the work performed to establish calibration traceability for the instrumentation used by Sandia National Laboratories at the Waste Isolation Pilot Plant (WIPP) during testing from 1980-1985. Identifying the calibration traceability is an important part of establishing a pedigree for the data and is part of the qualification of existing data. In general, the requirement states that the calibration of Measuring and Test equipment must have a valid relationship to nationally recognized standards or the basis for the calibration must be documented. Sandia recognized that just establishing calibration traceability would not necessarily mean that all QA requirements were met during the certification of test instrumentation. To address this concern, the assessment was expanded to include various activities.

  12. The GERDA calibration system

    Energy Technology Data Exchange (ETDEWEB)

    Baudis, Laura; Froborg, Francis; Tarka, Michael; Bruch, Tobias; Ferella, Alfredo [Physik-Institut, Universitaet Zuerich (Switzerland); Collaboration: GERDA-Collaboration

    2012-07-01

    A system with three identical custom made units is used for the energy calibration of the GERDA Ge diodes. To perform a calibration the {sup 228}Th sources are lowered from the parking positions at the top of the cryostat. Their positions are measured by two independent modules. One, the incremental encoder, counts the holes in the perforated steel band holding the sources, the other measures the drive shaft's angular position even if not powered. The system can be controlled remotely by a Labview program. The calibration data is analyzed by an iterative calibration algorithm determining the calibration functions for different energy reconstruction algorithms and the resolution of several peaks in the {sup 228}Th spectrum is determined. A Monte Carlo simulation using the GERDA simulation software MAGE has been performed to determine the background induced by the sources in the parking positions.

  13. Sandia WIPP calibration traceability

    International Nuclear Information System (INIS)

    Schuhen, M.D.; Dean, T.A.

    1996-05-01

    This report summarizes the work performed to establish calibration traceability for the instrumentation used by Sandia National Laboratories at the Waste Isolation Pilot Plant (WIPP) during testing from 1980-1985. Identifying the calibration traceability is an important part of establishing a pedigree for the data and is part of the qualification of existing data. In general, the requirement states that the calibration of Measuring and Test equipment must have a valid relationship to nationally recognized standards or the basis for the calibration must be documented. Sandia recognized that just establishing calibration traceability would not necessarily mean that all QA requirements were met during the certification of test instrumentation. To address this concern, the assessment was expanded to include various activities

  14. Development of an Efficient G-Quadruplex-Stabilised Thrombin-Binding Aptamer Containing a Three-Carbon Spacer Molecule

    DEFF Research Database (Denmark)

    Aaldering, Lukas J.; Poongavanam, Vasanthanathan; Langkjær, Niels

    2017-01-01

    The thrombin-binding aptamer (TBA), which shows anticoagulant properties, is one of the most studied G-quadruplex-forming aptamers. In this study, we investigated the impact of different chemical modifications such as a three-carbon spacer (spacer-C3), unlocked nucleic acid (UNA) and 3′-amino-mod...

  15. Alkylation of phosphorothioated thrombin binding aptamers improves the selectivity of inhibition of tumor cell proliferation upon anticoagulation.

    Science.gov (United States)

    Yang, Xiantao; Zhu, Yuejie; Wang, Chao; Guan, Zhu; Zhang, Lihe; Yang, Zhenjun

    2017-07-01

    Recently, aptamers have been extensively researched for therapy and diagnostic applications. Thrombin-binding aptamer is a 15nt deoxyribonucleic acid screened by SELEX, it can specifically bind to thrombin and inhibit blood coagulation. Since it is also endowed with excellent antitumor activity, the intrinsic anticoagulation advantage converted to a main potential side effect for its further application in antiproliferative therapy. Site-specific alkylation was conducted through nucleophilic reaction of phosphorothioated TBAs using bromide reagents. Circular dichroism (CD) spectroscopy and surface plasmon resonance (SPR) measurements were used to evaluate anticoagulation activity, and a CCK-8 assay was used to determine cell proliferation activity. The CD spectra of the modified TBAs were weakened, and their affinity for thrombin was dramatically reduced, as reflected by the K D values. On the other hand, their inhibition of A549 cells was retained. Incorporation of different alkyls apparently disrupted the binding of TBA to thrombin while maintaining the antitumor activity. A new modification strategy was established for the use of TBA as a more selective antitumor agent. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Effects of thrombin inhibition with melagatran on renal hemodynamics and function and liver integrity during early endotoxemia

    DEFF Research Database (Denmark)

    Nitescu, Nicoletta; Grimberg, Elisabeth; Ricksten, Sven-Erik

    2007-01-01

    Sepsis is associated with an activation of the coagulation system and multiorgan failure. The aim of the study was to examine the effects of selective thrombin inhibition with melagatran on renal hemodynamics and function, and liver integrity, during early endotoxemia. Endotoxemia was induced...

  17. Effect of the immobilisation of DNA aptamers on the detection of thrombin by means of surface plasmon resonance

    Czech Academy of Sciences Publication Activity Database

    Hianik, T.; Ostatná, V.; Vaisocherová, Hana; Homola, Jiří

    2008-01-01

    Roč. 391, č. 5 (2008), s. 1861-1869 ISSN 1618-2642 R&D Projects: GA AV ČR KAN200670701 Institutional research plan: CEZ:AV0Z20670512 Keywords : DNA aptamer * thrombin * dendrimer s Subject RIV: EI - Biotechnology ; Bionics Impact factor: 3.328, year: 2008

  18. Effects of hereditary and acquired risk factors of venous thrombosis on a thrombin generation-based APC resistance test

    NARCIS (Netherlands)

    Curvers, J; Thomassen, MCLGD; Rimmer, J; Hamulyak, K; van der Meer, J; Tans, G; Preston, FE; Rosing, J

    Background. Several hereditary and acquired risk factors for venous thromboembolism (VTE) are associated with impaired down-regulation of thrombin formation via the protein C pathway. To identify individuals at risk, functional tests are needed that estimate the risk to develop venous thrombosis.

  19. Graft Product for Autologous Peripheral Blood Stem Cell Transplantation Enhances Thrombin Generation and Expresses Procoagulant Microparticles and Tissue Factor.

    Science.gov (United States)

    Sidibe, Fatoumata; Spanoudaki, Anastasia; Vanneaux, Valerie; Mbemba, Elisabeth; Larghero, Jerome; Van Dreden, Patrick; Lotz, Jean-Pierre; Elalamy, Ismail; Larsen, Annette K; Gerotziafas, Grigoris T

    2018-05-01

    The beneficial effect of autologous peripheral blood stem cell transplantation (APBSCT) may be compromised by acute vascular complications related to hypercoagulability. We studied the impact of graft product on thrombin generation of normal plasma and the expression of tissue factor (TF) and procoagulant platelet-derived procoagulant microparticles (Pd-MPs) in samples of graft products. Graft products from 10 patients eligible for APBSCT were mixed with platelet-poor plasma (PPP) or platelet-rich plasma (PRP) from healthy volunteers and assessed for in vitro thrombin generation. In control experiments, thrombin generation was assessed in (1) PPP and PRP without any exogenous TF and/or procoagulant phospholipids, (2) PPP with the addition of TF (5 pM) and procoagulant phospholipids (4 μM), (3) in PRP with the addition of TF (5 pM). Graft products were assessed with Western blot assay for TF expression, with a specific clotting assay for TF activity and with flow cytometry assay for Pd-MPs. The graft product enhanced thrombin generation and its procoagulant activity was related to the presence of Pd-MPs and TF. The concentration of Pd-MPs in the graft product was characterized by a significant interindividual variability. The present study reveals the need for a thorough quality control of the graft products regarding their procoagulant potential.

  20. Binding characteristics of thrombin-activatable fibrinolysis inhibitor to streptococcal surface collagen-like proteins A and B

    NARCIS (Netherlands)

    Seron, Mercedes Valls; Plug, Tom; Marquart, J. Arnoud; Marx, Pauline F.; Herwald, Heiko; de Groot, Philip G.; Meijers, Joost C. M.

    2011-01-01

    Streptococcus pyogenes is the causative agent in a wide range of diseases in humans. Thrombin-activatable fibrinolysis inhibitor (TAFI) binds to collagen-like proteins ScIA and ScIB at the surface of S. pyogenes. Activation of TAFI at this surface redirects inflammation from a transient to chronic

  1. In vitro effect of hemodilution on activated clotting time and high-dose thrombin time during cardiopulmonary bypass

    NARCIS (Netherlands)

    Huyzen, RJ; vanOeveren, W; Wei, FY; Stellingwerf, P; Boonstra, PW; Gu, YJ

    Background. Extreme dilution of clotting factors, as may occur during pediatric or neonatal cardiopulmonary bypass, often leads to inadequate monitoring of anticoagulation with activated dotting time (ACT). In this study we postulate that the high-dose thrombin time (HiTT) is less influenced by

  2. Low cost automation

    International Nuclear Information System (INIS)

    1987-03-01

    This book indicates method of building of automation plan, design of automation facilities, automation and CHIP process like basics of cutting, NC processing machine and CHIP handling, automation unit, such as drilling unit, tapping unit, boring unit, milling unit and slide unit, application of oil pressure on characteristics and basic oil pressure circuit, application of pneumatic, automation kinds and application of process, assembly, transportation, automatic machine and factory automation.

  3. A machine vision system for the calibration of digital thermometers

    International Nuclear Information System (INIS)

    Vázquez-Fernández, Esteban; Dacal-Nieto, Angel; González-Jorge, Higinio; Alvarez-Valado, Victor; Martín, Fernando; Formella, Arno

    2009-01-01

    Automation is a key point in many industrial tasks such as calibration and metrology. In this context, machine vision has shown to be a useful tool for automation support, especially when there is no other option available. A system for the calibration of portable measurement devices has been developed. The system uses machine vision to obtain the numerical values shown by displays. A new approach based on human perception of digits, which works in parallel with other more classical classifiers, has been created. The results show the benefits of the system in terms of its usability and robustness, obtaining a success rate higher than 99% in display recognition. The system saves time and effort, and offers the possibility of scheduling calibration tasks without excessive attention by the laboratory technicians

  4. Automated Budget System -

    Data.gov (United States)

    Department of Transportation — The Automated Budget System (ABS) automates management and planning of the Mike Monroney Aeronautical Center (MMAC) budget by providing enhanced capability to plan,...

  5. A electrochemiluminescence aptasensor for detection of thrombin incorporating the capture aptamer labeled with gold nanoparticles immobilized onto the thio-silanized ITO electrode

    International Nuclear Information System (INIS)

    Fang Lanyun; Lue Zhaozi; Wei Hui; Wang Erkang

    2008-01-01

    A novel electrochemiluminescence (ECL) aptasensor was proposed for sensitive and cost-effective detection of the target thrombin adopted an aptamer-based sandwich format. To detect thrombin, capture aptamers labeled with gold nanoparticles (AuNPs) were first immobilized onto the thio-silanized ITO electrode surface through strong Au-S bonds. After catching the target thrombin, signal aptamers tagged with ECL labels were attached to the assembled electrode surface. As a result, an AuNPs-capture-aptamer/thrombin/ECL-tagged-signal-aptamer sandwich type was formed. Treating the resulting electrode surface with tri-n-propylamine (TPA) and applying a swept potential to the electrode, ECL response was generated which realized the detection of target protein. Spectroscopy and electrochemical impedance techniques were used to characterize and confirm the fabrication of the ECL aptasensor. AuNPs amplification and smart sensor fabrication art were implemented for the sensitive and cost-effective detection purpose. Signal-to-dose curve excellently followed a sandwich format equation and could be used to quantify the protein, and the detection limit was estimated to be 10 nM. Other forms of thrombin such as β- and γ-thrombins had negligible response, which indicated a high specificity of α-thrombin detection. The aptasensor opened up new fields of aptamer applications in ECL domain, a highly sensitive technique, and had a promising perspective to be applied in microarray analysis

  6. Rapid Upregulation of Orai1 Abundance in the Plasma Membrane of Platelets Following Activation with Thrombin and Collagen Related Peptide

    Directory of Open Access Journals (Sweden)

    Guilai Liu

    2015-11-01

    Full Text Available Background: Blood platelets accomplish primary hemostasis following vascular injury and contribute to the orchestration of occlusive vascular disease. Platelets are activated by an increase of cytosolic Ca2+-activity ([Ca2+]i, which is accomplished by Ca2+-release from intracellular stores and subsequent store operated Ca2+ entry (SOCE through Ca2+ release activated Ca2+ channel moiety Orai1. Powerful activators of platelets include thrombin and collagen related peptide (CRP, which are in part effective by activation of small G- protein Rac1. The present study explored the influence of thrombin and CRP on Orai1 protein abundance and cytosolic Ca2+-activity ([Ca2+]i in platelets drawn from wild type mice. Methods: Orai1 protein surface abundance was quantified utilizing CF™488A conjugated antibodies, and [Ca2+]i was determined with Fluo3-fluorescence. Results: In resting platelets, Orai1 protein abundance and [Ca2+]i were low. Thrombin (0.02 U/ml and CRP (5ug/ml within 2 min increased [Ca2+]i and Orai1 protein abundance at the platelet surface. [Ca2+]i was further increased by Ca2+ ionophore ionomycin (1 µM and by store depletion with the sarcoendoplasmatic Ca2+ ATPase inhibitor thapsigargin (1 µM. However, Orai1 protein abundance at the platelet surface was not significantly affected by ionomycin and only slightly increased by thapsigargin. The effect of thrombin and CRP on Orai1 abundance and [Ca2+]i was significantly blunted by Rac1 inhibitor NSC23766 (50 µM. Conclusion: The increase of [Ca2+]i following stimulation of platelets with thrombin and collagen related peptide is potentiated by ultrarapid Rac1 sensitive translocation of Orai1 into the cell membrane.

  7. Automation 2017

    CERN Document Server

    Zieliński, Cezary; Kaliczyńska, Małgorzata

    2017-01-01

    This book consists of papers presented at Automation 2017, an international conference held in Warsaw from March 15 to 17, 2017. It discusses research findings associated with the concepts behind INDUSTRY 4.0, with a focus on offering a better understanding of and promoting participation in the Fourth Industrial Revolution. Each chapter presents a detailed analysis of a specific technical problem, in most cases followed by a numerical analysis, simulation and description of the results of implementing the solution in a real-world context. The theoretical results, practical solutions and guidelines presented are valuable for both researchers working in the area of engineering sciences and practitioners looking for solutions to industrial problems. .

  8. Marketing automation

    Directory of Open Access Journals (Sweden)

    TODOR Raluca Dania

    2017-01-01

    Full Text Available The automation of the marketing process seems to be nowadays, the only solution to face the major changes brought by the fast evolution of technology and the continuous increase in supply and demand. In order to achieve the desired marketing results, businessis have to employ digital marketing and communication services. These services are efficient and measurable thanks to the marketing technology used to track, score and implement each campaign. Due to the technical progress, the marketing fragmentation, demand for customized products and services on one side and the need to achieve constructive dialogue with the customers, immediate and flexible response and the necessity to measure the investments and the results on the other side, the classical marketing approached had changed continue to improve substantially.

  9. Formation of PI 3-kinase products in platelets by thrombin, but not collagen, is dependent on synergistic autocrine stimulation, particularly through secreted ADP.

    Science.gov (United States)

    Selheim, F; Idsøe, R; Fukami, M H; Holmsen, H; Vassbotn, F S

    1999-10-05

    Platelet activation by thrombin or collagen results in secretion and synthesis of several platelet agonists that enhance the responses to the primary agonists (autocrine stimulation). To disclose the effects of thrombin and collagen on the phosphorylation of 3-phosphoinositides per se we incubated platelets with five inhibitors of platelet autocrine stimulation (IAS) that act extracellularly. We found that IAS almost totally blocked thrombin-induced production of phosphatidylinositol 3,4-bisphosphate [PtdIns(3,4)P(2)] and phosphatidylinositol 3,4,5-trisphosphate [PtdIns(3,4,5)P(3)]. In contrast, collagen induced massive production of PtdIns(3,4)P(2) and PtdIns(3,4,5)P(3) in the presence of IAS. When testing the effect of each inhibitor individually we found the strongest inhibition of thrombin-induced PtdIns(3,4)P(2) production with the ADP scavenger system CP/CPK. Furthermore, we found a strong synergistic effect between exogenously added ADP and thrombin on production of PtdIns(3,4)P(2). In contrast to the results from 3-phosphorylated phosphoinositides, CP/CPK had little effect on thrombin-induced protein tyrosine phosphorylation. Our results show the importance of autocrine stimulation in thrombin-induced accumulation of 3-phosphorylated phosphoinositides and raise the question as to whether thrombin by itself is capable of inducing PI 3-K activation. In marked contrast to thrombin, collagen per se appears to be able to trigger increased production of PtdIns(3,4)P(2) and PtdIns(3,4,5)P(3). Copyright 1999 Academic Press.

  10. iTRAQ quantitative proteomics-based identification of cell adhesion as a dominant phenotypic modulation in thrombin-stimulated human aortic endothelial cells.

    Science.gov (United States)

    Wang, Huang-Joe; Chen, Sung-Fang; Lo, Wan-Yu

    2015-05-01

    The phenotypic changes in thrombin-stimulated endothelial cells include alterations in permeability, cell shape, vasomotor tone, leukocyte trafficking, migration, proliferation, and angiogenesis. Previous studies regarding the pleotropic effects of thrombin on the endothelium used human umbilical vein endothelial cells (HUVECs)-cells derived from fetal tissue that does not exist in adults. Only a few groups have used screening approaches such as microarrays to profile the global effects of thrombin on endothelial cells. Moreover, the proteomic changes of thrombin-stimulated human aortic endothelial cells (HAECs) have not been elucidated. HAECs were stimulated with 2 units/mL thrombin for 5h and their proteome was investigated using isobaric tags for the relative and absolute quantification (iTRAQ) and the MetaCore(TM) software. A total of 627 (experiment A) and 622 proteins (experiment B) were quantified in the duplicated iTRAQ analyses. MetaCore(TM) pathway analysis identified cell adhesion as a dominant phenotype in thrombin-stimulated HAECs. Replicated iTRAQ data revealed that "Cell adhesion_Chemokines and adhesion," "Cell adhesion_Histamine H1 receptor signaling in the interruption of cell barrier integrity," and "Cell adhesion_Integrin-mediated cell adhesion and migration" were among the top 10 statistically significant pathways. The cell adhesion phenotype was verified by increased THP-1 adhesion to thrombin-stimulated HAECs. In addition, the expression of ICAM-1, VCAM-1, and SELE was significantly upregulated in thrombin-stimulated HAECs. Several regulatory pathways are altered in thrombin-stimulated HAECs, with cell adhesion being the dominant altered phenotype. Our findings show the feasibility of the iTRAQ technique for evaluating cellular responses to acute stimulation. Copyright © 2015 Elsevier Ltd. All rights reserved.

  11. A cluster of aspartic residues in the extracellular loop II of PAR 4 is important for thrombin interaction and activation of platelets.

    Science.gov (United States)

    Sánchez Centellas, Daniel; Gudlur, Sushanth; Vicente-Carrillo, Alejandro; Ramström, Sofia; Lindahl, Tomas L

    2017-06-01

    Thrombin activates platelets via proteolytic cleavage of protease-activated receptors (PARs) 1 and 4. The two PARs have distinct but complementary roles. The mechanisms responsible for PAR1 activation by thrombin have been extensively studied. However, much less is known regarding thrombin activation of PAR4, especially the potential involvement of regions of PAR4 other than the N-terminal, which is bound to the catalytic site of thrombin. We have studied PAR4 in S. cerevisiae strain MMY12, an expression system in which the GPCR receptors are connected to a Lac Z reporter gene resulting in increased β-galactosidase activity. This approach was used to assess PAR4 mutants to evaluate the contribution of different aspartic residues in facilitating PAR4 activation. Furthermore, peptides mimicking parts of the PAR4 N-terminal and the second extracellular loop (ECLII) were tested for their ability to inhibit platelet activation by thrombin. Binding of these peptides to γ-thrombin was studied by monitoring the decrease in tryptophan fluorescence intensity of thrombin. We conclude that not only the N-terminal but also the electronegative aspartic residues D224, D230 and D235 (located in ECLII) are be important for PAR4 binding to thrombin. We further suggest that they play a role for the tethered ligand binding to the receptor, as mutations also affected activation in response to a PAR4-activating peptide mimicking the new N-terminal formed after cleavage. This agrees with previous results on PAR1 and thrombin binding. We suggest that the ECLII of PAR4 could be a potential target for antithrombotic drug development. Copyright © 2017 Elsevier Ltd. All rights reserved.

  12. Compact radiometric microwave calibrator

    International Nuclear Information System (INIS)

    Fixsen, D. J.; Wollack, E. J.; Kogut, A.; Limon, M.; Mirel, P.; Singal, J.; Fixsen, S. M.

    2006-01-01

    The calibration methods for the ARCADE II instrument are described and the accuracy estimated. The Steelcast coated aluminum cones which comprise the calibrator have a low reflection while maintaining 94% of the absorber volume within 5 mK of the base temperature (modeled). The calibrator demonstrates an absorber with the active part less than one wavelength thick and only marginally larger than the mouth of the largest horn and yet black (less than -40 dB or 0.01% reflection) over five octaves in frequency

  13. Lidar to lidar calibration

    DEFF Research Database (Denmark)

    Fernandez Garcia, Sergio; Villanueva, Héctor

    This report presents the result of the lidar to lidar calibration performed for ground-based lidar. Calibration is here understood as the establishment of a relation between the reference lidar wind speed measurements with measurement uncertainties provided by measurement standard and corresponding...... lidar wind speed indications with associated measurement uncertainties. The lidar calibration concerns the 10 minute mean wind speed measurements. The comparison of the lidar measurements of the wind direction with that from the reference lidar measurements are given for information only....

  14. Investigation of the thrombin-generating capacity, evaluated by thrombogram, and clot formation evaluated by thrombelastography of platelets stored in the blood bank for up to 7 days

    DEFF Research Database (Denmark)

    Johansson, Per Ingemar; Svendsen, M.S.; Salado, J.

    2008-01-01

    thrombin) and endogenous thrombin potential (ETP; nm thrombin*min) were registered. Clot formation was evaluated by TEG and the R time (min), maxial amplitude (MA; mm), time to maximum thrombus generation (TMG; min) and maximum thrombus generation (MTG; dynes cm(-2) s(-1)) and total thrombus generation...... (TTG; dyne cm(-2)) were registered. RESULTS: Platelets become more procoagulant, evaluated both by TEG and CAT during storage. The reduction in CAT lag time and the ttPeak correlated with a decrease in the TEG R time and TMG (P

  15. PAR-1 and thrombin: the ties that bind the microenvironment to melanoma metastasis.

    Science.gov (United States)

    Zigler, Maya; Kamiya, Takafumi; Brantley, Emily C; Villares, Gabriel J; Bar-Eli, Menashe

    2011-11-01

    Progression of melanoma is dependent on cross-talk between tumor cells and the adjacent microenvironment. The thrombin receptor, protease-activated receptor-1 (PAR-1), plays a key role in exerting this function during melanoma progression. PAR-1 and its activating factors, which are expressed on tumor cells and the surrounding stroma, induce not only coagulation but also cell signaling, which promotes the metastatic phenotype. Several adhesion molecules, cytokines, growth factors, and proteases have recently been identified as downstream targets of PAR-1 and have been shown to modulate interactions between tumor cells and the microenvironment in the process of melanoma growth and metastasis. Inhibiting such interactions by targeting PAR-1 could potentially be a useful therapeutic modality for melanoma patients. ©2011 AACR.

  16. Historical perspective and contemporary management of acute coronary syndromes: from MONA to THROMBINS2.

    Science.gov (United States)

    Kline, Kristopher P; Conti, C Richard; Winchester, David E

    2015-01-01

    Acute coronary syndrome (ACS) remains a major burden on morbidity and mortality in the United States. Medical professionals and students often use the mnemonic 'MONA' (morphine, oxygen, nitroglycerin and aspirin) to recall treatments for ACS; however, this list of therapies is outdated. We provide a historical perspective on 'MONA,' attempt to uncover its origin in the medical literature, and demonstrate the myriad changes that have occurred over the last 50 years of ACS management. We have developed a novel mnemonic, 'THROMBINS2' (thienopyridines, heparin/enoxaparin, renin-angiotensin system blockers, oxygen, morphine, beta blocker, intervention, nitroglycerin, statin/salicylate) to help bedside clinicians recall all the elements of contemporary ACS management. We demonstrate the mortality benefit for each component of contemporary ACS management, correlating the continued improvement with historical data on mortality after myocardial infarction. We encourage providers to utilize this mnemonic to explore options and guide treatments in ACS patients.

  17. Thrombin-activatable fibrinolysis inhibitor activity in healthy and diseased dogs

    DEFF Research Database (Denmark)

    Jessen, Lisbeth Rem; Wiinberg, Bo; Kjelgaard-Hansen, Mads

    2010-01-01

    Background: In people, increased thrombin-activatable fibrinolysis inhibitor (TAFI) antigen has been associated with increased risk of thrombosis, and decreased TAFI may contribute to bleeding diathesis. TAFI activity in dogs has been described in experimental models, but not in dogs...... with spontaneous disease. Objective: The aim of this study was to compare TAFI activity in healthy dogs with TAFI activity in dogs with spontaneous disease. Methods: Plasma samples from 20 clinically healthy Beagles and from 35 dogs with various diseases were analyzed using a commercial chromogenic assay...... that measured TAFI activity relative to activity in standardized pooled human plasma. Results: Median TAFI activity for the 20 Beagles was 46.1% (range 32.2-70.8%) compared with 62.6% (29.1-250%) for the 35 diseased dogs, and 14/35 (40%) had TAFI activities >the upper limit for controls. The highest individual...

  18. The Calibration Home Base for Imaging Spectrometers

    Directory of Open Access Journals (Sweden)

    Johannes Felix Simon Brachmann

    2016-08-01

    Full Text Available The Calibration Home Base (CHB is an optical laboratory designed for the calibration of imaging spectrometers for the VNIR/SWIR wavelength range. Radiometric, spectral and geometric calibration as well as the characterization of sensor signal dependency on polarization are realized in a precise and highly automated fashion. This allows to carry out a wide range of time consuming measurements in an ecient way. The implementation of ISO 9001 standards in all procedures ensures a traceable quality of results. Spectral measurements in the wavelength range 380–1000 nm are performed to a wavelength uncertainty of +- 0.1 nm, while an uncertainty of +-0.2 nm is reached in the wavelength range 1000 – 2500 nm. Geometric measurements are performed at increments of 1.7 µrad across track and 7.6 µrad along track. Radiometric measurements reach an absolute uncertainty of +-3% (k=1. Sensor artifacts, such as caused by stray light will be characterizable and correctable in the near future. For now, the CHB is suitable for the characterization of pushbroom sensors, spectrometers and cameras. However, it is planned to extend the CHBs capabilities in the near future such that snapshot hyperspectral imagers can be characterized as well. The calibration services of the CHB are open to third party customers from research institutes as well as industry.

  19. A guide to automation techniques in calorimetry

    International Nuclear Information System (INIS)

    Renz, D.P.; Wetzel, J.R.; Breakall, K.L.; James, S.J.; Kasperski, P.W.

    1992-01-01

    Many improvements occurring in calorimetry measurement technology in the past few years will help current users of calorimeters to achieve better use of their existing systems. These include a more user friendly operator computer interface, a more detailed printout at the end of each run, improved data processing to eliminate operator error, and improved system monitoring to detect system or environmental problems. In addition, an electrical calibration heater has been developed to replace plutonium-238 heat standards for calibrating calorimeters, and several automation systems allow for easier and safer system operation

  20. Comparison of Ultrasound-Guided Thrombin Injection of Iatrogenic Pseudoaneurysms Based on Neck Dimension.

    Science.gov (United States)

    Yang, Ethan Y; Tabbara, Marwan M; Sanchez, Priscila G; Abi-Chaker, Andrew M; Patel, Jaimin; Bornak, Arash; Jones, Keith M; Rey, Jorge

    2018-02-01

    Ultrasound-guided thrombin injection (UGTI) of femoral artery pseudoaneurysms after endovascular procedures is an effective therapy. There is controversy in the literature regarding injecting pseudoaneurysms with short and/or wide necks. This article reports our experience in UGTI of pseudoaneurysms in 1 hospital regarding the efficacy of this treatment in all pseudoaneurysms regardless of the size of the necks. A retrospective review of 46 patients diagnosed between 2011 and 2016 with groin pseudoaneurysms using established duplex ultrasound criteria. Mean age was 68 years (range 27-87). Ten pseudoaneurysms thrombosed spontaneously, 5 were thrombosed by ultrasound-guided compression, and 2 were treated surgically due to disqualifying criteria. In this retrospective review, we analyzed the remaining 29 pseudoaneurysms regarding the dimensions of their neck lengths and outcomes after attempting thrombin injection. The mean aneurysm neck length and width were 1.03 ± 0.9 cm and 0.30 ± 0.1 cm, respectively. All 29 patients were evaluated with respect to pseudoaneurysm size, neck length, neck width, and complexity. Successful treatment of 29 pseudoaneurysms (2 external iliac, 20 common femoral, 2 deep femoral, and 5 superficial femoral) with UGTI was achieved without complications in 100% of the cases, regardless of pseudoaneurysm size, neck dimensions, or complexity. Anticoagulation status did not affect the efficacy of the procedure. Nine of the 29 pseudoaneurysms (31.0%) had neck length less than 0.5 cm. This study demonstrates the safety and efficacy of UGTI in treating iatrogenic pseudoaneurysm in 29 of 29 patients, even in patients with pseudoaneurysm with short neck lengths. Our experiences support injecting all pseudoaneurysms irrespective of dimension. Copyright © 2017 Elsevier Inc. All rights reserved.

  1. Construction of a Bivalent Thrombin Binding Aptamer and Its Antidote with Improved Properties

    Directory of Open Access Journals (Sweden)

    Quintin W. Hughes

    2017-10-01

    Full Text Available Aptamers are short synthetic DNA or RNA oligonucleotides that adopt secondary and tertiary conformations based on Watson–Crick base-pairing interactions and can be used to target a range of different molecules. Two aptamers, HD1 and HD22, that bind to exosites I and II of the human thrombin molecule, respectively, have been extensively studied due to their anticoagulant potentials. However, a fundamental issue preventing the clinical translation of many aptamers is degradation by nucleases and reduced pharmacokinetic properties requiring higher dosing regimens more often. In this study, we have chemically modified the design of previously described thrombin binding aptamers targeting exosites I, HD1, and exosite II, HD22. The individual aptamers were first modified with an inverted deoxythymidine nucleotide, and then constructed bivalent aptamers by connecting the HD1 and HD22 aptamers either through a triethylene glycol (TEG linkage or four consecutive deoxythymidines together with an inverted deoxythymidine nucleotide at the 3′-end. The anticoagulation potential, the reversal of coagulation with different antidote sequences, and the nuclease stability of the aptamers were then investigated. The results showed that a bivalent aptamer RNV220 containing an inverted deoxythymidine and a TEG linkage chemistry significantly enhanced the anticoagulation properties in blood plasma and nuclease stability compared to the existing aptamer designs. Furthermore, a bivalent antidote sequence RNV220AD efficiently reversed the anticoagulation effect of RNV220 in blood plasma. Based on our results, we believe that RNV220 could be developed as a potential anticoagulant therapeutic molecule.

  2. Characterization of Ixophilin, a thrombin inhibitor from the gut of Ixodes scapularis.

    Directory of Open Access Journals (Sweden)

    Sukanya Narasimhan

    Full Text Available Ixodes scapularis, the black-legged tick, vectors several human pathogens including Borrelia burgdorferi, the agent of Lyme disease in North America. Pathogen transmission to the vertebrate host occurs when infected ticks feed on the mammalian host to obtain a blood meal. Efforts to understand how the tick confronts host hemostatic mechanisms and imbibes a fluid blood meal have largely focused on the anticoagulation strategies of tick saliva. The blood meal that enters the tick gut remains in a fluid state for several days during the process of feeding, and the role of the tick gut in maintaining the blood-meal fluid is not understood. We now demonstrate that the tick gut produces a potent inhibitor of thrombin, a key enzyme in the mammalian coagulation cascade. Chromatographic fractionation of engorged tick gut proteins identified one predominant thrombin inhibitory activity associated with an approximately 18 kDa protein, henceforth referred to as Ixophilin. The ixophilin gene was preferentially transcribed in the guts of feeding nymphs. Expression began after 24 hours of feeding, coincident with the flow of host blood into the tick gut. Immunity against Ixophilin delayed tick feeding, and decreased feeding efficiency significantly. Surprisingly, immunity against Ixophilin resulted in increased Borrelia burgdorferi transmission to the host, possibly due to delayed feeding and increased transmission opportunity. These observations illuminate the potential drawbacks of targeting individual tick proteins in a functional suite. They also underscore the need to identify the "anticoagulome" of the tick gut, and to prioritize a critical subset of anticoagulants that could be targeted to efficiently thwart tick feeding, and block pathogen transmission to the vertebrate host.

  3. Effects of the oral, direct thrombin inhibitor dabigatran on five common coagulation assays.

    Science.gov (United States)

    Lindahl, Tomas L; Baghaei, Fariba; Blixter, Inger Fagerberg; Gustafsson, Kerstin M; Stigendal, Lennart; Sten-Linder, Margareta; Strandberg, Karin; Hillarp, Andreas

    2011-02-01

    Dabigatran is an oral, reversible thrombin inhibitor that has shown promising results in large clinical trials. Laboratory monitoring is not needed but the effects on common coagulation assays are incompletely known. Dabigatran was added to plasma from healthy subjects in the concentration range 0-1,000 μg/l and analysed using several reagents for activated thromboplastin time (APTT), prothrombin time (PT), fibrinogen, antithrombin, and activated protein C resistance. Typical trough concentrations are about 50 μg/l, peak concentrations 100-300 μg/l. At 100 μg/l all APTT-results were prolonged. The concentration required to double APTT ranged between 227 and 286 μg/l, the responses for all five reagents were similar. PT-reagents were much less affected with almost no samples above INR 1.2 at 100 μg/l. The effect was sample dilution dependent with PT Quick type more sensitive than PT Owren type methods. If a patient on dabigatran has prolonged APTT, >90 seconds, and Quick PT INR>2 or Owren PT INR>1.5 over-dosing or accumulation of dabigatran should be considered. Two of four fibrinogen reagents underestimated the fibrinogen concentration considerably at expected peak concentration. Methods based on inhibition of thrombin over-estimated the antithrombin concentration, but not Xa-based. The APC-resistance methods over-estimated the APC-ratio, which may lead to miss-classification of factor V Leiden patients as being normal. Different coagulation assays, and even different reagents within an assay group, display variable effects at therapeutic concentrations of dabigatran. Some of these assay variations are of clinical importance, thus knowledge is needed for a correct interpretation of results.

  4. Air Data Calibration Facility

    Data.gov (United States)

    Federal Laboratory Consortium — This facility is for low altitude subsonic altimeter system calibrations of air vehicles. Mission is a direct support of the AFFTC mission. Postflight data merge is...

  5. SRHA calibration curve

    Data.gov (United States)

    U.S. Environmental Protection Agency — an UV calibration curve for SRHA quantitation. This dataset is associated with the following publication: Chang, X., and D. Bouchard. Surfactant-Wrapped Multiwalled...

  6. BES online calibration system

    International Nuclear Information System (INIS)

    Zhang Bingyun; Li Xiaonan; Zhu Kejun; Zhang Jiawen; Gong Mingyu

    2003-01-01

    We constructed BES (Beijing Spectrometer) online calibration system to ensure the coherence of readout electronic channels due to huge data volume in high energy physics experiment. This paper describes the structure of hardware and software, and its characteristic and function

  7. Calibrated Properties Model

    International Nuclear Information System (INIS)

    Ahlers, C.F.; Liu, H.H.

    2001-01-01

    The purpose of this Analysis/Model Report (AMR) is to document the Calibrated Properties Model that provides calibrated parameter sets for unsaturated zone (UZ) flow and transport process models for the Yucca Mountain Site Characterization Project (YMP). This work was performed in accordance with the AMR Development Plan for U0035 Calibrated Properties Model REV00 (CRWMS M and O 1999c). These calibrated property sets include matrix and fracture parameters for the UZ Flow and Transport Model (UZ Model), drift seepage models, drift-scale and mountain-scale coupled-processes models, and Total System Performance Assessment (TSPA) models as well as Performance Assessment (PA) and other participating national laboratories and government agencies. These process models provide the necessary framework to test conceptual hypotheses of flow and transport at different scales and predict flow and transport behavior under a variety of climatic and thermal-loading conditions

  8. Calibrated Properties Model

    International Nuclear Information System (INIS)

    Ahlers, C.; Liu, H.

    2000-01-01

    The purpose of this Analysis/Model Report (AMR) is to document the Calibrated Properties Model that provides calibrated parameter sets for unsaturated zone (UZ) flow and transport process models for the Yucca Mountain Site Characterization Project (YMP). This work was performed in accordance with the ''AMR Development Plan for U0035 Calibrated Properties Model REV00. These calibrated property sets include matrix and fracture parameters for the UZ Flow and Transport Model (UZ Model), drift seepage models, drift-scale and mountain-scale coupled-processes models, and Total System Performance Assessment (TSPA) models as well as Performance Assessment (PA) and other participating national laboratories and government agencies. These process models provide the necessary framework to test conceptual hypotheses of flow and transport at different scales and predict flow and transport behavior under a variety of climatic and thermal-loading conditions

  9. SPOTS Calibration Example

    Directory of Open Access Journals (Sweden)

    Patterson E.

    2010-06-01

    Full Text Available The results are presented using the procedure outlined by the Standardisation Project for Optical Techniques of Strain measurement to calibrate a digital image correlation system. The process involves comparing the experimental data obtained with the optical measurement system to the theoretical values for a specially designed specimen. The standard states the criteria which must be met in order to achieve successful calibration, in addition to quantifying the measurement uncertainty in the system. The system was evaluated at three different displacement load levels, generating strain ranges from 289 µstrain to 2110 µstrain. At the 289 µstrain range, the calibration uncertainty was found to be 14.1 µstrain, and at the 2110 µstrain range it was found to be 28.9 µstrain. This calibration procedure was performed without painting a speckle pattern on the surface of the metal. Instead, the specimen surface was prepared using different grades of grit paper to produce the desired texture.

  10. Scanner calibration revisited

    Directory of Open Access Journals (Sweden)

    Pozhitkov Alexander E

    2010-07-01

    Full Text Available Abstract Background Calibration of a microarray scanner is critical for accurate interpretation of microarray results. Shi et al. (BMC Bioinformatics, 2005, 6, Art. No. S11 Suppl. 2. reported usage of a Full Moon BioSystems slide for calibration. Inspired by the Shi et al. work, we have calibrated microarray scanners in our previous research. We were puzzled however, that most of the signal intensities from a biological sample fell below the sensitivity threshold level determined by the calibration slide. This conundrum led us to re-investigate the quality of calibration provided by the Full Moon BioSystems slide as well as the accuracy of the analysis performed by Shi et al. Methods Signal intensities were recorded on three different microarray scanners at various photomultiplier gain levels using the same calibration slide from Full Moon BioSystems. Data analysis was conducted on raw signal intensities without normalization or transformation of any kind. Weighted least-squares method was used to fit the data. Results We found that initial analysis performed by Shi et al. did not take into account autofluorescence of the Full Moon BioSystems slide, which led to a grossly distorted microarray scanner response. Our analysis revealed that a power-law function, which is explicitly accounting for the slide autofluorescence, perfectly described a relationship between signal intensities and fluorophore quantities. Conclusions Microarray scanners respond in a much less distorted fashion than was reported by Shi et al. Full Moon BioSystems calibration slides are inadequate for performing calibration. We recommend against using these slides.

  11. Calibration of thermoluminiscent materials

    International Nuclear Information System (INIS)

    Bos, A.J.J.

    1989-07-01

    In this report the relation between exposure and absorbed radiation dose in various materials is represented, on the base of recent data. With the help of this a calibration procedure for thermoluminescent materials, adapted to the IRI radiation standard is still the exposure in rontgen. In switching to the air kerma standard the calibration procedure will have to be adapted. (author). 6 refs.; 4 tabs

  12. Both Automation and Paper.

    Science.gov (United States)

    Purcell, Royal

    1988-01-01

    Discusses the concept of a paperless society and the current situation in library automation. Various applications of automation and telecommunications are addressed, and future library automation is considered. Automation at the Monroe County Public Library in Bloomington, Indiana, is described as an example. (MES)

  13. Calibrating nacelle lidars

    Energy Technology Data Exchange (ETDEWEB)

    Courtney, M.

    2013-01-15

    Nacelle mounted, forward looking wind lidars are beginning to be used to provide reference wind speed measurements for the power performance testing of wind turbines. In such applications, a formal calibration procedure with a corresponding uncertainty assessment will be necessary. This report presents four concepts for performing such a nacelle lidar calibration. Of the four methods, two are found to be immediately relevant and are pursued in some detail. The first of these is a line of sight calibration method in which both lines of sight (for a two beam lidar) are individually calibrated by accurately aligning the beam to pass close to a reference wind speed sensor. A testing procedure is presented, reporting requirements outlined and the uncertainty of the method analysed. It is seen that the main limitation of the line of sight calibration method is the time required to obtain a representative distribution of radial wind speeds. An alternative method is to place the nacelle lidar on the ground and incline the beams upwards to bisect a mast equipped with reference instrumentation at a known height and range. This method will be easier and faster to implement and execute but the beam inclination introduces extra uncertainties. A procedure for conducting such a calibration is presented and initial indications of the uncertainties given. A discussion of the merits and weaknesses of the two methods is given together with some proposals for the next important steps to be taken in this work. (Author)

  14. Laser guided automated calibrating system for accurate bracket ...

    African Journals Online (AJOL)

    It is widely recognized that accurate bracket placement is of critical importance in the efficient application of biomechanics and in realizing the full potential of a preadjusted edgewise appliance. Aim: The purpose of ... placement. Keywords: Hough transforms, Indirect bonding technique, Laser, Orthodontic bracket placement ...

  15. AUTOMATED CALIBRATION OF FEM MODELS USING LIDAR POINT CLOUDS

    Directory of Open Access Journals (Sweden)

    B. Riveiro

    2018-05-01

    Full Text Available In present work it is pretended to estimate elastic parameters of beams through the combined use of precision geomatic techniques (laser scanning and structural behaviour simulation tools. The study has two aims, on the one hand, to develop an algorithm able to interpret automatically point clouds acquired by laser scanning systems of beams subjected to different load situations on experimental tests; and on the other hand, to minimize differences between deformation values given by simulation tools and those measured by laser scanning. In this way we will proceed to identify elastic parameters and boundary conditions of structural element so that surface stresses can be estimated more easily.

  16. Laser Guided Automated Calibrating System for Accurate Bracket ...

    African Journals Online (AJOL)

    Background: The basic premise of preadjusted bracket system is accurate bracket positioning. ... using MATLAB ver. 7 software (The MathWorks Inc.). These images are in the form of matrices of size 640 × 480. 650 nm (red light) type III diode laser is used as ... motion control and Pitch, Yaw, Roll degrees of freedom (DOF).

  17. Automated Vision Test Development and Validation

    Science.gov (United States)

    2016-11-01

    crystal display monitor (NEC Multisync, P232W) at 1920x1080 resolution. Proper calibration was confirmed using a spot photometer/colorimeter (X-Rite i1...visual input to the right and left eye was achieved using liquid crystal display shuttered glasses (NVIDIA 3D Vision 2). The stereo target (Figure 4...threshold on the automated tasks. • Subjects had a lower (better) threshold on color testing for all cone types using the OCCT due to a ceiling

  18. Measurement quality assurance for beta particle calibrations at NIST

    International Nuclear Information System (INIS)

    Soares, C.G.; Pruitt, J.S.

    1993-01-01

    Standardized beta-particle fields have been established in an international standard and have been adopted for use in several U.S. dosimeter and instrument testing standards. Calibration methods and measurement quality assurance procedures employed at the National Institute of Standards and Technology (NIST) for beta-particle calibrations in these reference fields are discussed. The calibration facility including the NIST-automated extrapolation ionization chamber is described, and some sample results of calibrations are shown. Methods for establishing and maintaining traceability to NIST of secondary laboratories are discussed. Currently, there are problems in finding a good method for routine testing of traceability to NIST. Some examples of past testing methods are given and solutions to this problem are proposed

  19. Automatic Calibration of High Density Electric Muscle Stimulation

    DEFF Research Database (Denmark)

    Knibbe, Jarrod; Strohmeier, Paul; Boring, Sebastian

    2017-01-01

    . (2) EMS requires time consuming, expert calibration -- confining these interaction techniques to the lab. EMS arrays have been shown to increase stimulation resolution, but as calibration complexity increases exponentially as more electrodes are used, we require heuristics or automated procedures......Electric muscle stimulation (EMS) can enable mobile force feedback, support pedestrian navigation, or confer object affordances. To date, however, EMS is limited by two interlinked problems. (1) EMS is low resolution -- achieving only coarse movements and constraining opportunities for exploration...... for successful calibration. We explore the feasibility of using electromyography (EMG) to auto-calibrate high density EMS arrays. We determine regions of muscle activity during human-performed gestures, to inform stimulation patterns for EMS-performed gestures. We report on a study which shows that auto...

  20. Measurement quality assurance for beta particle calibrations at NIST

    Energy Technology Data Exchange (ETDEWEB)

    Soares, C.G.; Pruitt, J.S. [National Institute of Standards and Technology, Gaithersburg, MD (United States)

    1993-12-31

    Standardized beta-particle fields have been established in an international standard and have been adopted for use in several U.S. dosimeter and instrument testing standards. Calibration methods and measurement quality assurance procedures employed at the National Institute of Standards and Technology (NIST) for beta-particle calibrations in these reference fields are discussed. The calibration facility including the NIST-automated extrapolation ionization chamber is described, and some sample results of calibrations are shown. Methods for establishing and maintaining traceability to NIST of secondary laboratories are discussed. Currently, there are problems in finding a good method for routine testing of traceability to NIST. Some examples of past testing methods are given and solutions to this problem are proposed.

  1. Electrochemical and circular dichroism spectroscopic evidence of two types of interaction between [Ru(NH3)(6)](3+) and an elongated thrombin binding aptamer G-quadruplex

    Czech Academy of Sciences Publication Activity Database

    De Rache, A.; Kejnovská, Iva; Buess-Herman, C.; Doneux, T.

    2015-01-01

    Roč. 179, OCT 2015 (2015), s. 84-92 ISSN 0013-4686 Institutional support: RVO:68081707 Keywords : Biosensors * Thrombin binding aptamer * Hexaammineruthenium Subject RIV: BO - Biophysics Impact factor: 4.803, year: 2015

  2. I trust it, but I don't know why: effects of implicit attitudes toward automation on trust in an automated system.

    Science.gov (United States)

    Merritt, Stephanie M; Heimbaugh, Heather; LaChapell, Jennifer; Lee, Deborah

    2013-06-01

    This study is the first to examine the influence of implicit attitudes toward automation on users' trust in automation. Past empirical work has examined explicit (conscious) influences on user level of trust in automation but has not yet measured implicit influences. We examine concurrent effects of explicit propensity to trust machines and implicit attitudes toward automation on trust in an automated system. We examine differential impacts of each under varying automation performance conditions (clearly good, ambiguous, clearly poor). Participants completed both a self-report measure of propensity to trust and an Implicit Association Test measuring implicit attitude toward automation, then performed an X-ray screening task. Automation performance was manipulated within-subjects by varying the number and obviousness of errors. Explicit propensity to trust and implicit attitude toward automation did not significantly correlate. When the automation's performance was ambiguous, implicit attitude significantly affected automation trust, and its relationship with propensity to trust was additive: Increments in either were related to increases in trust. When errors were obvious, a significant interaction between the implicit and explicit measures was found, with those high in both having higher trust. Implicit attitudes have important implications for automation trust. Users may not be able to accurately report why they experience a given level of trust. To understand why users trust or fail to trust automation, measurements of implicit and explicit predictors may be necessary. Furthermore, implicit attitude toward automation might be used as a lever to effectively calibrate trust.

  3. Thrombin has biphasic effects on the nitric oxide-cGMP pathway in endothelial cells and contributes to experimental pulmonary hypertension.

    Directory of Open Access Journals (Sweden)

    Katrin F Nickel

    Full Text Available BACKGROUND: A potential role for coagulation factors in pulmonary arterial hypertension has been recently described, but the mechanism of action is currently not known. Here, we investigated the interactions between thrombin and the nitric oxide-cGMP pathway in pulmonary endothelial cells and experimental pulmonary hypertension. PRINCIPAL FINDINGS: Chronic treatment with the selective thrombin inhibitor melagatran (0.9 mg/kg daily via implanted minipumps reduced right ventricular hypertrophy in the rat monocrotaline model of experimental pulmonary hypertension. In vitro, thrombin was found to have biphasic effects on key regulators of the nitric oxide-cGMP pathway in endothelial cells (HUVECs. Acute thrombin stimulation led to increased expression of the cGMP-elevating factors endothelial nitric oxide synthase (eNOS and soluble guanylate cyclase (sGC subunits, leading to increased cGMP levels. By contrast, prolonged exposition of pulmonary endothelial cells to thrombin revealed a characteristic pattern of differential expression of the key regulators of the nitric oxide-cGMP pathway, in which specifically the factors contributing to cGMP elevation (eNOS and sGC were reduced and the cGMP-hydrolyzing PDE5 was elevated (qPCR and Western blot. In line with the differential expression of key regulators of the nitric oxide-cGMP pathway, a reduction of cGMP by prolonged thrombin stimulation was found. The effects of prolonged thrombin exposure were confirmed in endothelial cells of pulmonary origin (HPAECs and HPMECs. Similar effects could be induced by activation of protease-activated receptor-1 (PAR-1. CONCLUSION: These findings suggest a link between thrombin generation and cGMP depletion in lung endothelial cells through negative regulation of the nitric oxide-cGMP pathway, possibly mediated via PAR-1, which could be of relevance in pulmonary arterial hypertension.

  4. An electrochemical aptasensor based on TiO2/MWCNT and a novel synthesized Schiff base nanocomposite for the ultrasensitive detection of thrombin.

    Science.gov (United States)

    Heydari-Bafrooei, Esmaeil; Amini, Maryam; Ardakani, Mehdi Hatefi

    2016-11-15

    A sensitive aptasensor based on a robust nanocomposite of titanium dioxide nanoparticles, multiwalled carbon nanotubes (MWCNT), chitosan and a novel synthesized Schiff base (SB) (TiO2/MWCNT/CHIT/SB) on the surface of a glassy carbon electrode (GCE) was developed for thrombin detection. The resultant nanocomposite can provide a large surface area, excellent electrocatalytic activity, and high stability, which would improve immobilization sites for biological molecules, allow remarkable amplification of the electrochemical signal and contribute to improved sensitivity. Thrombin aptamers were simply immobilized onto the TiO2-MWCNT/CHIT-SB nanocomposite matrix through simple π - π stacking and electrostatic interactions between CHIT/SB and aptamer strands. The electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV) and differential pulse voltammetry (DPV) were used to analyze the surface characterization of unmodified GCE and TiO2-MWCNT/CHIT-SB modified GCE, and also the interaction between aptamer and thrombin. In the presence of thrombin, the aptamer on the adsorbent layer captures the target on the electrode interface, which makes a barrier for electrons and inhibits electron transfer, thereby resulting in decreased DPV and increased impedance signals of the TiO2-MWCNT/CHIT-SB modified GCE. Furthermore, the proposed aptasensor has a very low LOD of 1.0fmolL(-1) thrombin within the detection range of 0.00005-10nmolL(-1). The aptasensor also presents high specificity and reproducibility for thrombin, which is unaffected by the coexistence of other proteins. Clinical application was performed with analysis of the thrombin levels in blood and CSF samples obtained from patients with MS, Parkinson, Epilepsy and Polyneuropathy using both the aptasensor and commercial ELISA kit. The results revealed the proposed system to be a promising candidate for clinical analysis of thrombin. Copyright © 2016 Elsevier B.V. All rights reserved.

  5. Molecular design and structure--activity relationships leading to the potent, selective, and orally active thrombin active site inhibitor BMS-189664.

    Science.gov (United States)

    Das, Jagabandhu; Kimball, S David; Hall, Steven E; Han, Wen Ching; Iwanowicz, Edwin; Lin, James; Moquin, Robert V; Reid, Joyce A; Sack, John S; Malley, Mary F; Chang, Chiehying Y; Chong, Saeho; Wang-Iverson, David B; Roberts, Daniel G M; Seiler, Steven M; Schumacher, William A; Ogletree, Martin L

    2002-01-07

    A series of structurally novel small molecule inhibitors of human alpha-thrombin was prepared to elucidate their structure-activity relationships (SARs), selectivity and activity in vivo. BMS-189664 (3) is identified as a potent, selective, and orally active reversible inhibitor of human alpha-thrombin which is efficacious in vivo in a mouse lethality model, and at inhibiting both arterial and venous thrombosis in cynomolgus monkey models.

  6. POLCAL - POLARIMETRIC RADAR CALIBRATION

    Science.gov (United States)

    Vanzyl, J.

    1994-01-01

    Calibration of polarimetric radar systems is a field of research in which great progress has been made over the last few years. POLCAL (Polarimetric Radar Calibration) is a software tool intended to assist in the calibration of Synthetic Aperture Radar (SAR) systems. In particular, POLCAL calibrates Stokes matrix format data produced as the standard product by the NASA/Jet Propulsion Laboratory (JPL) airborne imaging synthetic aperture radar (AIRSAR). POLCAL was designed to be used in conjunction with data collected by the NASA/JPL AIRSAR system. AIRSAR is a multifrequency (6 cm, 24 cm, and 68 cm wavelength), fully polarimetric SAR system which produces 12 x 12 km imagery at 10 m resolution. AIRSTAR was designed as a testbed for NASA's Spaceborne Imaging Radar program. While the images produced after 1991 are thought to be calibrated (phase calibrated, cross-talk removed, channel imbalance removed, and absolutely calibrated), POLCAL can and should still be used to check the accuracy of the calibration and to correct it if necessary. Version 4.0 of POLCAL is an upgrade of POLCAL version 2.0 released to AIRSAR investigators in June, 1990. New options in version 4.0 include automatic absolute calibration of 89/90 data, distributed target analysis, calibration of nearby scenes with calibration parameters from a scene with corner reflectors, altitude or roll angle corrections, and calibration of errors introduced by known topography. Many sources of error can lead to false conclusions about the nature of scatterers on the surface. Errors in the phase relationship between polarization channels result in incorrect synthesis of polarization states. Cross-talk, caused by imperfections in the radar antenna itself, can also lead to error. POLCAL reduces cross-talk and corrects phase calibration without the use of ground calibration equipment. Removing the antenna patterns during SAR processing also forms a very important part of the calibration of SAR data. Errors in the

  7. Vibration transducer calibration techniques

    Science.gov (United States)

    Brinkley, D. J.

    1980-09-01

    Techniques for the calibration of vibration transducers used in the Aeronautical Quality Assurance Directorate of the British Ministry of Defence are presented. Following a review of the types of measurements necessary in the calibration of vibration transducers, the performance requirements of vibration transducers, which can be used to measure acceleration, velocity or vibration amplitude, are discussed, with particular attention given to the piezoelectric accelerometer. Techniques for the accurate measurement of sinusoidal vibration amplitude in reference-grade transducers are then considered, including the use of a position sensitive photocell and the use of a Michelson laser interferometer. Means of comparing the output of working-grade accelerometers with that of previously calibrated reference-grade devices are then outlined, with attention given to a method employing a capacitance bridge technique and a method to be used at temperatures between -50 and 200 C. Automatic calibration procedures developed to speed up the calibration process are outlined, and future possible extensions of system software are indicated.

  8. Calibration Under Uncertainty.

    Energy Technology Data Exchange (ETDEWEB)

    Swiler, Laura Painton; Trucano, Timothy Guy

    2005-03-01

    This report is a white paper summarizing the literature and different approaches to the problem of calibrating computer model parameters in the face of model uncertainty. Model calibration is often formulated as finding the parameters that minimize the squared difference between the model-computed data (the predicted data) and the actual experimental data. This approach does not allow for explicit treatment of uncertainty or error in the model itself: the model is considered the %22true%22 deterministic representation of reality. While this approach does have utility, it is far from an accurate mathematical treatment of the true model calibration problem in which both the computed data and experimental data have error bars. This year, we examined methods to perform calibration accounting for the error in both the computer model and the data, as well as improving our understanding of its meaning for model predictability. We call this approach Calibration under Uncertainty (CUU). This talk presents our current thinking on CUU. We outline some current approaches in the literature, and discuss the Bayesian approach to CUU in detail.

  9. Thrombin generation assay as a possible tool for assessment of reduced activity of clotting factors induced by antiphospholipid antibodies and in-vitro evaluation of treatment options.

    Science.gov (United States)

    Livnat, Tami; Zivelin, Ariella; Tamarin, Ilia; Guetta, Victor; Salomon, Ophira

    2009-12-01

    Bleeding is a rare manifestation of antiphospholipid syndrome, unless associated with reduced clotting factors or severe thrombocytopenia. Accurate assessment of the autoantibodies in plasma is very important since the autoantibodies can lead to bleeding or thrombosis. The objective of the present study was to define the inhibitors causing reduced clotting activity in a patient with antiphospholipids antibodies and to assess the potential of thrombin generation assay to assist in establishment of optimal treatment in case of major bleeding. Levels of clotting factors as well as inhibitors to factors II, V, VII, VIII, IX, X and XI were defined. For detection of inhibitors to prothrombin crossed immunoelectrophoresis was used. IgG was purified by commercial protein A column. Thrombin generation was measured using a fluorometric assay in platelet-poor and platelet-rich plasma. Inhibitors toward the activity of factors V, VII, VIII, IX, X and XI were defined and also an inhibitor to prothrombin antigen. No thrombin generation was induced in the patient's plasma by recalcification even in the presence of recombinant factor VIIa or factor VIII inhibitor bypassing activity. In contrast, addition of platelets from either donor or patient or synthetic phospholipids normalized the thrombin generation. The thrombin generation model showed that the addition of platelets and no recombinant factor VIIa or factor VIII inhibitor bypassing activity would correct thrombin generation in vitro. On this basis, platelet concentrates were administered to a patient with bleeding caused by lupus anticoagulant and low clotting factors activity.

  10. Fluorescence enhancement upon G-quadruplex folding: synthesis, structure, and biophysical characterization of a dansyl/cyclodextrin-tagged thrombin binding aptamer.

    Science.gov (United States)

    De Tito, Stefano; Morvan, François; Meyer, Albert; Vasseur, Jean-Jacques; Cummaro, Annunziata; Petraccone, Luigi; Pagano, Bruno; Novellino, Ettore; Randazzo, Antonio; Giancola, Concetta; Montesarchio, Daniela

    2013-11-20

    A novel fluorescent thrombin binding aptamer (TBA), conjugated with the environmentally sensitive dansyl probe at the 3'-end and a β-cyclodextrin residue at the 5'-end, has been efficiently synthesized exploiting Cu(I)-catalyzed azide-alkyne cycloaddition procedures. Its conformation and stability in solution have been studied by an integrated approach, combining in-depth NMR, CD, fluorescence, and DSC studies. ITC measurements have allowed us to analyze in detail its interaction with human thrombin. All the collected data show that this bis-conjugated aptamer fully retains its G-quadruplex formation ability and thrombin recognition properties, with the terminal appendages only marginally interfering with the conformational behavior of TBA. Folding of this modified aptamer into the chairlike, antiparallel G-quadruplex structure, promoted by K(+) and/or thrombin binding, typical of TBA, is associated with a net fluorescence enhancement, due to encapsulation of dansyl, attached at the 3'-end, into the apolar cavity of the β-cyclodextrin at the 5'-end. Overall, the structural characterization of this novel, bis-conjugated TBA fully demonstrates its potential as a diagnostic tool for thrombin recognition, also providing a useful basis for the design of suitable aptamer-based devices for theranostic applications, allowing simultaneously both detection and inhibition or modulation of the thrombin activity.

  11. Toehold strand displacement-driven assembly of G-quadruplex DNA for enzyme-free and non-label sensitive fluorescent detection of thrombin.

    Science.gov (United States)

    Xu, Yunying; Zhou, Wenjiao; Zhou, Ming; Xiang, Yun; Yuan, Ruo; Chai, Yaqin

    2015-02-15

    Based on a new signal amplification strategy by the toehold strand displacement-driven cyclic assembly of G-quadruplex DNA, the development of an enzyme-free and non-label aptamer sensing approach for sensitive fluorescent detection of thrombin is described. The target thrombin associates with the corresponding aptamer of the partial dsDNA probes and liberates single stranded initiation sequences, which trigger the toehold strand displacement assembly of two G-quadruplex containing hairpin DNAs. This toehold strand displacement reaction leads to the cyclic reuse of the initiation sequences and the production of DNA assemblies with numerous G-quadruplex structures. The fluorescent dye, N-Methyl mesoporphyrin IX, binds to these G-quadruplex structures and generates significantly amplified fluorescent signals to achieve highly sensitive detection of thrombin down to 5 pM. Besides, this method shows high selectivity towards the target thrombin against other control proteins. The developed thrombin sensing method herein avoids the modification of the probes and the involvement of any enzyme or nanomaterial labels for signal amplification. With the successful demonstration for thrombin detection, our approach can be easily adopted to monitor other target molecules in a simple, low-cost, sensitive and selective way by choosing appropriate aptamer/ligand pairs. Copyright © 2014 Elsevier B.V. All rights reserved.

  12. Flow cytometry analysis reveals different activation profiles of thrombin- or TRAP-stimulated platelets in db/db mice. The regulatory role of PAR-3.

    Science.gov (United States)

    Kassassir, Hassan; Siewiera, Karolina; Talar, Marcin; Przygodzki, Tomasz; Watala, Cezary

    2017-06-01

    Recent studies have shown that it may be the concentration of thrombin, which is discriminative in determining of the mechanism of platelet activation via protease activated receptors (PARs). Whether the observed phenomenon of differentiated responses of mouse platelets to various thrombin concentrations in non-diabetic db/+ and diabetic db/db mice depends upon the concerted action of various PARs, remains to be established. We found elevated reactivity of platelets, as well as the enhanced PAR-3 expression in response to both the used concentrations of AYPGKF in db/db mice, as compared to db/+ heterozygotes. At low concentration of thrombin platelets from diabetic mice demonstrated hyperreactivity, reflected by higher expression of PAR-3. For higher thrombin concentration, blood platelets from db/db mice appeared hyporeactive, compared to db/+ animals, while no significant differences in PAR-3 expression were observed between diabetic and non-diabetic mice. The novel and previously unreported finding resulting from our study is that the increased expression of PAR-3 in response to either TRAP for PAR-4 or low thrombin (when PAR-4 is not the efficient thrombin receptor) may be one of the key events contributing to higher reactivity of platelets in db/db mice. Copyright © 2017 Elsevier Inc. All rights reserved.

  13. Secretory products from thrombin-stimulated human platelets exert an inhibitory effect on NK-cytotoxic activity

    DEFF Research Database (Denmark)

    Skov Madsen, P; Hokland, P; Hokland, M

    1987-01-01

    We have investigated the interaction between human platelets and the NK-system, with special emphasis on the action of secretory products from platelets in an NK assay with 51Cr-labelled K562 as target cells. Supernatants from thrombin-stimulated platelets added to the NK assay consistently...... decreased the NK-cytotoxicity by 40% +/- 4.3%, indicating the existence of secreted products from platelets as a source of NK-inhibiting substances. In contrast, no direct cytotoxic effect of these secretory products on the target cells (K562) was seen. Thus, normal human platelets, when stimulated...... with thrombin, are capable of secreting different, yet undefined factors, which significantly inhibit NK activity in vitro. The results also suggest that the role of products from contaminating in vitro activated platelets should be borne in mind when performing conventional NK assays. Udgivelsesdato: 1986-Oct...

  14. Ibis ground calibration

    International Nuclear Information System (INIS)

    Bird, A.J.; Barlow, E.J.; Tikkanen, T.; Bazzano, A.; Del Santo, M.; Ubertini, P.; Blondel, C.; Laurent, P.; Lebrun, F.; Di Cocco, G.; Malaguti, E.; Gabriele, M.; La Rosa, G.; Segreto, A.; Quadrini, E.; Volkmer, R.

    2003-01-01

    We present an overview of results obtained from IBIS ground calibrations. The spectral and spatial characteristics of the detector planes and surrounding passive materials have been determined through a series of calibration campaigns. Measurements of pixel gain, energy resolution, detection uniformity, efficiency and imaging capability are presented. The key results obtained from the ground calibration have been: - optimization of the instrument tunable parameters, - determination of energy linearity for all detection modes, - determination of energy resolution as a function of energy through the range 20 keV - 3 MeV, - demonstration of imaging capability in each mode, - measurement of intrinsic detector non-uniformity and understanding of the effects of passive materials surrounding the detector plane, and - discovery (and closure) of various leakage paths through the passive shielding system

  15. Gamma counter calibration system

    International Nuclear Information System (INIS)

    1977-01-01

    A method and apparatus are described for the calibration of a gamma radiation measurement instrument to be used over any of a number of different absolute energy ranges. The method includes the steps of adjusting the overall signal gain associated with pulses which are derived from detected gamma rays, until the instrument is calibrated for a particular absolute energy range; then storing parameter settings corresponding to the adjusted overall signal gain, and repeating the process for other desired absolute energy ranges. The stored settings can be subsequently retrieved and reapplied so that test measurements can be made using a selected one of the absolute energy ranges. Means are provided for adjusting the overall signal gain and a specific technique is disclosed for making coarse, then fine adjustments to the signal gain, for rapid convergence of the required calibration settings. (C.F.)

  16. Individual dosimetry and calibration

    International Nuclear Information System (INIS)

    Hoefert, M.; Nielsen, M.

    1996-01-01

    In 1995 both the Individual Dosimetry and Calibration Sections worked under the condition of a status quo and concentrated fully on the routine part of their work. Nevertheless, the machine for printing the bar code which will be glued onto the film holder and hence identify the people when entering into high radiation areas was put into operation and most of the holders were equipped with the new identification. As far as the Calibration Section is concerned the project of the new source control system that is realized by the Technical Support Section was somewhat accelerated

  17. Radiation Calibration Measurements

    International Nuclear Information System (INIS)

    Omondi, C.

    2017-01-01

    KEBS Radiation Dosimetry mandate are: Custodian of Kenya Standards on Ionizing radiation, Ensure traceability to International System (SI ) and Calibration radiation equipment. RAF 8/040 on Radioisotope applications for troubleshooting and optimizing industrial process established Radiotracer Laboratory objective is to introduce and implement radiotracer technique for problem solving of industrial challenges. Gamma ray scanning technique applied is to Locate blockages, Locate liquid in vapor lines, Locate areas of lost refractory or lining in a pipe and Measure flowing densities. Equipment used for diagnostic and radiation protection must be calibrated to ensure Accuracy and Traceability

  18. Calibrating Legal Judgments

    Directory of Open Access Journals (Sweden)

    Frederick Schauer

    2017-09-01

    Full Text Available Objective to study the notion and essence of legal judgments calibration the possibilities of using it in the lawenforcement activity to explore the expenses and advantages of using it. Methods dialectic approach to the cognition of social phenomena which enables to analyze them in historical development and functioning in the context of the integrity of objective and subjective factors it determined the choice of the following research methods formallegal comparative legal sociological methods of cognitive psychology and philosophy. Results In ordinary life people who assess other peoplersaquos judgments typically take into account the other judgments of those they are assessing in order to calibrate the judgment presently being assessed. The restaurant and hotel rating website TripAdvisor is exemplary because it facilitates calibration by providing access to a raterrsaquos previous ratings. Such information allows a user to see whether a particular rating comes from a rater who is enthusiastic about every place she patronizes or instead from someone who is incessantly hard to please. And even when less systematized as in assessing a letter of recommendation or college transcript calibration by recourse to the decisional history of those whose judgments are being assessed is ubiquitous. Yet despite the ubiquity and utility of such calibration the legal system seems perversely to reject it. Appellate courts do not openly adjust their standard of review based on the previous judgments of the judge whose decision they are reviewing nor do judges in reviewing legislative or administrative decisions magistrates in evaluating search warrant representations or jurors in assessing witness perception. In most legal domains calibration by reference to the prior decisions of the reviewee is invisible either because it does not exist or because reviewing bodies are unwilling to admit using what they in fact know and employ. Scientific novelty for the first

  19. Calibration of scanning Lidar

    DEFF Research Database (Denmark)

    Gómez Arranz, Paula; Courtney, Michael

    This report describes the tests carried out on a scanning lidar at the DTU Test Station for large wind turbines, Høvsøre. The tests were divided in two parts. In the first part, the purpose was to obtain wind speed calibrations at two heights against two cup anemometers mounted on a mast. Additio......This report describes the tests carried out on a scanning lidar at the DTU Test Station for large wind turbines, Høvsøre. The tests were divided in two parts. In the first part, the purpose was to obtain wind speed calibrations at two heights against two cup anemometers mounted on a mast...

  20. Anti-thrombin III, Protein C, and Protein S deficiency in acute coronary syndrome

    Directory of Open Access Journals (Sweden)

    Dasnan Ismail

    2002-06-01

    Full Text Available The final most common pathway for the majority of coronary artery disease is occlusion of a coronary vessel. Under normal conditions, antithrombin III (AT III, protein C, and protein S as an active protein C cofactor, are natural anticoagulants (hemostatic control that balances procoagulant activity (thrombin antithrombin complex balance to prevent thrombosis. If the condition becomes unbalanced, natural anticoagulants and the procoagulants can lead to thrombosis. Thirty subjects with acute coronary syndrome (ACS were studied for the incidence of antithrombin III (AT III, protein C, and protein S deficiencies, and the result were compare to the control group. Among patients with ACS, the frequency of distribution of AT-III with activity < 75% were 23,3% (7 of 30, and only 6,7% ( 2 of 30 in control subject. No one of the 30 control subject have protein C activity deficient, in ACS with activity < 70% were 13,3% (4 of 30. Fifteen out of the 30 (50% control subjects had protein S activity deficiency, while protein S deficiency activity < 70% was found 73.3.% (22 out of 30. On linear regression, the deterministic coefficient of AT-III activity deficiency to the development ACS was 13,25 %, and the deterministic coefficient of protein C activity deficient to the development of ACS was 9,06 %. The cut-off point for AT-III without protein S deficiency expected to contribute to the development of vessel disease was 45%. On discriminant analysis, protein C activity deficiency posed a risk for ACS of 4,5 greater than non deficient subjects, and AT-III activity deficiency posed a risk for ACS of 3,5 times greater than non deficient subjects. On binary logistic regression, protein S activity acted only as a reinforcing factor of AT-III activity deficiency in the development of ACS. Protein C and AT III deficiency can trigger ACS, with determinant coefficients of 9,06% and 13,25% respectively. Low levels of protein C posed a greater risk of

  1. Combination of FVIII and by-passing agent potentiates in vitro thrombin production in haemophilia A inhibitor plasma.

    Science.gov (United States)

    Klintman, Jenny; Astermark, Jan; Berntorp, Erik

    2010-11-01

    The by-passing agents, recombinant activated factor VII (rFVIIa) and activated prothrombin complex concentrate (APCC), are important tools in the treatment of patients with haemophilia A and high-responding inhibitory antibodies. It has been observed clinically that in some patients undergoing immune tolerance induction the bleeding frequency decreases, hypothetically caused by a transient haemostatic effect of infused FVIII not measurable ex vivo. We evaluated how by-passing agents and factor VIII (FVIII) affect thrombin generation (TG) in vitro using plasma from 11 patients with severe haemophilia A and high titre inhibitors. Samples were spiked with combinations of APCC, rFVIIa and five different FVIII products. Combination of APCC and FVIII showed a synergistic effect in eliciting TG (Pproducts. When rFVIIa and FVIII were combined the interaction between the preparations was found to be additive. APCC and rFVIIa were then combined without FVIII, resulting in an additive effect on thrombin production. Each product separately increased TG above baseline. In conclusion, the amount of thrombin formed in vitro by adding a by-passing agent, was higher in the presence of FVIII. Our findings support the use of FVIII in by-passing therapy to optimize the haemostatic effect. © 2010 Blackwell Publishing Ltd.

  2. A sensitive electrochemical aptasensor based on water soluble CdSe quantum dots (QDs) for thrombin determination

    Energy Technology Data Exchange (ETDEWEB)

    Li Yanfen; Han Min [Jiangsu Laboratory of New Power Batteries, Jiangsu Key Laboratory of Biofuctional Materials, College of Chemistry and Materials Science, Nanjing Normal University, Nanjing 210097 (China); Bai Hongyan [Jiangsu Laboratory of New Power Batteries, Jiangsu Key Laboratory of Biofuctional Materials, College of Chemistry and Materials Science, Nanjing Normal University, Nanjing 210097 (China); College of Biological and Chemical Engineering, Jiaxing College, Jiaxing 314001 (China); Wu Yong [Jiangsu Laboratory of New Power Batteries, Jiangsu Key Laboratory of Biofuctional Materials, College of Chemistry and Materials Science, Nanjing Normal University, Nanjing 210097 (China); Dai Zhihui, E-mail: daizhihuii@njnu.edu.cn [Jiangsu Laboratory of New Power Batteries, Jiangsu Key Laboratory of Biofuctional Materials, College of Chemistry and Materials Science, Nanjing Normal University, Nanjing 210097 (China); Bao Jianchun, E-mail: baojianchun@njnu.edu.cn [Jiangsu Laboratory of New Power Batteries, Jiangsu Key Laboratory of Biofuctional Materials, College of Chemistry and Materials Science, Nanjing Normal University, Nanjing 210097 (China)

    2011-08-01

    A novel aptamer biosensor with easy operation and good sensitivity, specificity, stability and reproducibility was developed by immobilizing the aptamer on water soluble CdSe quantum dots (QDs) modified on the top of the glassy carbon electrode (GCE). Methylene blue (MB) was intercalated into the aptamer sequence and used as an electrochemical marker. CdSe QDs improved the electrochemical signal because of their larger surface area and ion centers of CdSe QDs may also had a major role on amplifying the signal. The higher ion concentration caused more combination of aptamer which caused larger signal. The thrombin was detected by differential pulse voltammetry (DPV) quantitatively. Under optimal conditions, the two linear ranges were obtained from 3 to 13 {mu}g mL{sup -1} and from 14 to 31 {mu}g mL{sup -1}, respectively. The detection limit was 0.08 {mu}g mL{sup -1} at 3{sigma}. The constructed biosensor had better responses compared with that in the absence of the CdSe QDs immobilizing. The control experiment was also carried out by using BSA, casein and IgG in the absence of thrombin. The results showed that the aptasensor had good specificity, stability and reproducibility to the thrombin. Moreover, the aptasensor could be used for detection of real sample with consistent results in comparison with those obtained by fluorescence method which could provide a promising platform for fabrication of aptamer based biosensors.

  3. ECAL Energy Flow Calibration

    CERN Multimedia

    CERN. Geneva

    2015-01-01

    My talk will be covering my work as a whole over the course of the semester. The focus will be on using energy flow calibration in ECAL to check the precision of the corrections made by the light monitoring system used to account for transparency loss within ECAL crystals due to radiation damage over time.

  4. Calibration with Absolute Shrinkage

    DEFF Research Database (Denmark)

    Øjelund, Henrik; Madsen, Henrik; Thyregod, Poul

    2001-01-01

    In this paper, penalized regression using the L-1 norm on the estimated parameters is proposed for chemometric je calibration. The algorithm is of the lasso type, introduced by Tibshirani in 1996 as a linear regression method with bound on the absolute length of the parameters, but a modification...

  5. Calibration bench of flowmeters

    International Nuclear Information System (INIS)

    Bremond, J.; Da Costa, D.; Calvet, A.; Vieuxmaire, C.

    1966-01-01

    This equipment is devoted to the comparison of signals from two turbines installed in the Cabri experimental loop. The signal is compared to the standard turbine. The characteristics and the performance of the calibration bench are presented. (A.L.B.)

  6. Calibration of farmer dosemeters

    International Nuclear Information System (INIS)

    Ahmad, S.S.; Anwar, K.; Arshed, W.; Mubarak, M.A.; Orfi, S.D.

    1984-08-01

    The Farmer Dosemeters of Atomic Energy Medical Centre (AEMC) Jamshoro were calibrated in the Secondary Standard Dosimetry Laboratory (SSDL) at PINSTECH, using the NPL Secondary Standard Therapy level X-ray exposure meter. The results are presented in this report. (authors)

  7. Physiotherapy ultrasound calibrations

    International Nuclear Information System (INIS)

    Gledhill, M.

    1996-01-01

    Calibration of physiotherapy ultrasound equipment has long been a problem. Numerous surveys around the world over the past 20 years have all found that only a low percentage of the units tested had an output within 30% of that indicatd. In New Zealand, a survey carried out by the NRL in 1985 found that only 24% had an output, at the maximum setting, within + or - 20% of that indicated. The present performance Standard for new equipment (NZS 3200.2.5:1992) requires that the measured output should not deviate from that indicated by more than + or - 30 %. This may be tightened to + or - 20% in the next few years. Any calibration is only as good as the calibration equipment. Some force balances can be tested with small weights to simulate the force exerted by an ultrasound beam, but with others this is not possible. For such balances, testing may only be feasible with a calibrated source which could be used like a transfer standard. (author). 4 refs., 3 figs

  8. NVLAP calibration laboratory program

    Energy Technology Data Exchange (ETDEWEB)

    Cigler, J.L.

    1993-12-31

    This paper presents an overview of the progress up to April 1993 in the development of the Calibration Laboratories Accreditation Program within the framework of the National Voluntary Laboratory Accreditation Program (NVLAP) at the National Institute of Standards and Technology (NIST).

  9. NVLAP calibration laboratory program

    International Nuclear Information System (INIS)

    Cigler, J.L.

    1993-01-01

    This paper presents an overview of the progress up to April 1993 in the development of the Calibration Laboratories Accreditation Program within the framework of the National Voluntary Laboratory Accreditation Program (NVLAP) at the National Institute of Standards and Technology (NIST)

  10. Gamma ray calibration system

    International Nuclear Information System (INIS)

    Rosauer, P.J.; Flaherty, J.J.

    1981-01-01

    This invention is in the field of gamma ray inspection devices for tubular products and the like employing an improved calibrating block which prevents the sensing system from being overloaded when no tubular product is present, and also provides the operator with a means for visually detecting the presence of wall thicknesses which are less than a required minimum. (author)

  11. An automated swimming respirometer

    DEFF Research Database (Denmark)

    STEFFENSEN, JF; JOHANSEN, K; BUSHNELL, PG

    1984-01-01

    An automated respirometer is described that can be used for computerized respirometry of trout and sharks.......An automated respirometer is described that can be used for computerized respirometry of trout and sharks....

  12. Autonomy and Automation

    Science.gov (United States)

    Shively, Jay

    2017-01-01

    A significant level of debate and confusion has surrounded the meaning of the terms autonomy and automation. Automation is a multi-dimensional concept, and we propose that Remotely Piloted Aircraft Systems (RPAS) automation should be described with reference to the specific system and task that has been automated, the context in which the automation functions, and other relevant dimensions. In this paper, we present definitions of automation, pilot in the loop, pilot on the loop and pilot out of the loop. We further propose that in future, the International Civil Aviation Organization (ICAO) RPAS Panel avoids the use of the terms autonomy and autonomous when referring to automated systems on board RPA. Work Group 7 proposes to develop, in consultation with other workgroups, a taxonomy of Levels of Automation for RPAS.

  13. Configuration Management Automation (CMA) -

    Data.gov (United States)

    Department of Transportation — Configuration Management Automation (CMA) will provide an automated, integrated enterprise solution to support CM of FAA NAS and Non-NAS assets and investments. CMA...

  14. A critical comparison of systematic calibration protocols for activated sludge models: a SWOT analysis.

    Science.gov (United States)

    Sin, Gürkan; Van Hulle, Stijn W H; De Pauw, Dirk J W; van Griensven, Ann; Vanrolleghem, Peter A

    2005-07-01

    Modelling activated sludge systems has gained an increasing momentum after the introduction of activated sludge models (ASMs) in 1987. Application of dynamic models for full-scale systems requires essentially a calibration of the chosen ASM to the case under study. Numerous full-scale model applications have been performed so far which were mostly based on ad hoc approaches and expert knowledge. Further, each modelling study has followed a different calibration approach: e.g. different influent wastewater characterization methods, different kinetic parameter estimation methods, different selection of parameters to be calibrated, different priorities within the calibration steps, etc. In short, there was no standard approach in performing the calibration study, which makes it difficult, if not impossible, to (1) compare different calibrations of ASMs with each other and (2) perform internal quality checks for each calibration study. To address these concerns, systematic calibration protocols have recently been proposed to bring guidance to the modeling of activated sludge systems and in particular to the calibration of full-scale models. In this contribution four existing calibration approaches (BIOMATH, HSG, STOWA and WERF) will be critically discussed using a SWOT (Strengths, Weaknesses, Opportunities, Threats) analysis. It will also be assessed in what way these approaches can be further developed in view of further improving the quality of ASM calibration. In this respect, the potential of automating some steps of the calibration procedure by use of mathematical algorithms is highlighted.

  15. PLEIADES ABSOLUTE CALIBRATION : INFLIGHT CALIBRATION SITES AND METHODOLOGY

    Directory of Open Access Journals (Sweden)

    S. Lachérade

    2012-07-01

    Full Text Available In-flight calibration of space sensors once in orbit is a decisive step to be able to fulfil the mission objectives. This article presents the methods of the in-flight absolute calibration processed during the commissioning phase. Four In-flight calibration methods are used: absolute calibration, cross-calibration with reference sensors such as PARASOL or MERIS, multi-temporal monitoring and inter-bands calibration. These algorithms are based on acquisitions over natural targets such as African deserts, Antarctic sites, La Crau (Automatic calibration station and Oceans (Calibration over molecular scattering or also new extra-terrestrial sites such as the Moon and selected stars. After an overview of the instrument and a description of the calibration sites, it is pointed out how each method is able to address one or several aspects of the calibration. We focus on how these methods complete each other in their operational use, and how they help building a coherent set of information that addresses all aspects of in-orbit calibration. Finally, we present the perspectives that the high level of agility of PLEIADES offers for the improvement of its calibration and a better characterization of the calibration sites.

  16. Mercury CEM Calibration

    Energy Technology Data Exchange (ETDEWEB)

    John F. Schabron; Joseph F. Rovani; Susan S. Sorini

    2007-03-31

    The Clean Air Mercury Rule (CAMR) which was published in the Federal Register on May 18, 2005, requires that calibration of mercury continuous emissions monitors (CEMs) be performed with NIST-traceable standards. Western Research Institute (WRI) is working closely with the Electric Power Research Institute (EPRI), the National Institute of Standards and Technology (NIST), and the Environmental Protection Agency (EPA) to facilitate the development of the experimental criteria for a NIST traceability protocol for dynamic elemental mercury vapor generators. The traceability protocol will be written by EPA. Traceability will be based on the actual analysis of the output of each calibration unit at several concentration levels ranging from about 2-40 ug/m{sup 3}, and this analysis will be directly traceable to analyses by NIST using isotope dilution inductively coupled plasma/mass spectrometry (ID ICP/MS) through a chain of analyses linking the calibration unit in the power plant to the NIST ID ICP/MS. Prior to this project, NIST did not provide a recommended mercury vapor pressure equation or list mercury vapor pressure in its vapor pressure database. The NIST Physical and Chemical Properties Division in Boulder, Colorado was subcontracted under this project to study the issue in detail and to recommend a mercury vapor pressure equation that the vendors of mercury vapor pressure calibration units can use to calculate the elemental mercury vapor concentration in an equilibrium chamber at a particular temperature. As part of this study, a preliminary evaluation of calibration units from five vendors was made. The work was performed by NIST in Gaithersburg, MD and Joe Rovani from WRI who traveled to NIST as a Visiting Scientist.

  17. Does Lipid Profile Affect Thrombin Generation During Ramadan Fasting in Patients With Cardiovascular Risks?

    Science.gov (United States)

    Sassi, Mouna; Chakroun, Taher; Chouchène, Saoussen; Hellara, Ilhem; Boubaker, Hamdi; Grissa, Mohamed Habib; Khochtali, Ines; Hassine, Mohsen; Addad, Faouzi; Elalamy, Ismail; Nouira, Semir

    2017-11-01

    There is evidence that diet and variation in lipid metabolism can influence blood coagulation, but little is known about the effect of Ramadan fasting on plasmatic coagulation pattern. We investigated the effect of Ramadan fasting on thrombin generation (TG) in patients with cardiovascular disease (CVD) risks, and we aimed to assess the effect of lipid profile on TG parameters. The study was conducted in 36 adults having at least 2 CVD risks and in 30 healthy controls. Coagulation pattern was assessed by both classical clotting times and TG test. A complete lipid profile was performed simultaneously. Patients were invited 2 times: 1 week before Ramadan and during the last week of the Ramadan. The TG parameters were not different in patients with CVD risks compared to healthy controls. Fasting had no effect on plasmatic coagulation parameters and on TG profile. Individual analysis of the mean rate index (MRI) of TG revealed 3 groups: group 1 with no modification of MRI, group 2 with a significant increase in MRI (81.64 nM/min vs 136.07 nM/min; P fasting did not influence the global coagulation pattern in patients with CVD risks. Whereas, a significant increase in the propagation phase of TG was associated with a significant increase in cholesterol levels, which was not found with the other TG parameters.

  18. Noncoded amino acids in protein engineering: Structure-activity relationship studies of hirudin-thrombin interaction.

    Science.gov (United States)

    De Filippis, Vincenzo; Acquasaliente, Laura; Pontarollo, Giulia; Peterle, Daniele

    2018-01-01

    The advent of recombinant DNA technology allowed to site-specifically insert, delete, or mutate almost any amino acid in a given protein, significantly improving our knowledge of protein structure, stability, and function. Nevertheless, a quantitative description of the physical and chemical basis that makes a polypeptide chain to efficiently fold into a stable and functionally active conformation is still elusive. This mainly originates from the fact that nature combined, in a yet unknown manner, different properties (i.e., hydrophobicity, conformational propensity, polarizability, and hydrogen bonding capability) into the 20 standard natural amino acids, thus making difficult, if not impossible, to univocally relate the change in protein stability or function to the alteration of physicochemical properties caused by amino acid exchange(s). In this view, incorporation of noncoded amino acids with tailored side chains, allowing to finely tune the structure at a protein site, would facilitate to dissect the effects of a given mutation in terms of one or a few physicochemical properties, thus much expanding the scope of physical organic chemistry in the study of proteins. In this review, relevant applications from our laboratory will be presented on the use of noncoded amino acids in structure-activity relationships studies of hirudin binding to thrombin. © 2017 International Union of Biochemistry and Molecular Biology, Inc.

  19. Dynamic affinity chromatography in the separation of sulfated lignins binding to thrombin

    Science.gov (United States)

    Liang, Aiye; Thakkar, Jay N.; Hindle, Michael; Desai, Umesh R.

    2013-01-01

    Sulfated low molecular weight lignins (LMWLs), a mixture of chemo-enzymatically prepared oligomers, have been found to be potent antagonists of coagulation. However, structures that induce anticoagulation remain unidentified. The highly polar sulfate groups on these molecules and the thousands of different structures present in these mixtures make traditional chromatographic resolution of sulfated LMWLs difficult. We performed dynamic thrombin affinity chromatography monitored using chromogenic substrate hydrolysis assay to isolate sulfated LMWL fractions that differed significantly in their biophysical and biochemical properties. Three fractions, I35, I55 and Peak II, were isolated from the starting complex mixture. Independent plasma clotting assays suggested that I35 possessed good anticoagulation potential (APTT = 4.2 μM; PT = 6.8 μM), while I55 and Peak II were approximately 10- and 100-fold less potent. The ESI-MS spectrum of this oligomeric fraction showed multiple peaks at 684.8, 610.6, 557.4, 541.4, 536.5, and 519.4 m/z, which most probably arise from variably functionalized (β-O4—β-β-linked trimers and/or a β-O4—β-O4-linked dimers. The first direct observation of these structures in sulfated LMWLs will greatly assist in the discovery of more potent sulfated LMWL-based anticoagulants. PMID:23122400

  20. Unfolding mechanism of thrombin-binding aptamer revealed by molecular dynamics simulation and Markov State Model.

    Science.gov (United States)

    Zeng, Xiaojun; Zhang, Liyun; Xiao, Xiuchan; Jiang, Yuanyuan; Guo, Yanzhi; Yu, Xinyan; Pu, Xuemei; Li, Menglong

    2016-04-05

    Thrombin-binding aptamer (TBA) with the sequence 5'GGTTGGTGTGGTTGG3' could fold into G-quadruplex, which correlates with functionally important genomic regionsis. However, unfolding mechanism involved in the structural stability of G-quadruplex has not been satisfactorily elucidated on experiments so far. Herein, we studied the unfolding pathway of TBA by a combination of molecular dynamics simulation (MD) and Markov State Model (MSM). Our results revealed that the unfolding of TBA is not a simple two-state process but proceeds along multiple pathways with multistate intermediates. One high flux confirms some observations from NMR experiment. Another high flux exhibits a different and simpler unfolding pathway with less intermediates. Two important intermediate states were identified. One is similar to the G-triplex reported in the folding of G-quadruplex, but lack of H-bonding between guanines in the upper plane. More importantly, another intermediate state acting as a connector to link the folding region and the unfolding one, was the first time identified, which exhibits higher population and stability than the G-triplex-like intermediate. These results will provide valuable information for extending our understanding the folding landscape of G-quadruplex formation.

  1. CALIBRATION PROCEDURES ON OBLIQUE CAMERA SETUPS

    Directory of Open Access Journals (Sweden)

    G. Kemper

    2016-06-01

    step with the help of the nadir camera and the GPS/IMU data, an initial orientation correction and radial correction were calculated. With this approach, the whole project was calculated and calibrated in one step. During the iteration process the radial and tangential parameters were switched on individually for the camera heads and after that the camera constants and principal point positions were checked and finally calibrated. Besides that, the bore side calibration can be performed either on basis of the nadir camera and their offsets, or independently for each camera without correlation to the others. This must be performed in a complete mission anyway to get stability between the single camera heads. Determining the lever arms of the nodal-points to the IMU centre needs more caution than for a single camera especially due to the strong tilt angle. Prepared all these previous steps, you get a highly accurate sensor that enables a fully automated data extraction with a rapid update of you existing data. Frequently monitoring urban dynamics is then possible in fully 3D environment.

  2. Field calibration of cup anemometers

    DEFF Research Database (Denmark)

    Schmidt Paulsen, Uwe; Mortensen, Niels Gylling; Hansen, Jens Carsten

    2007-01-01

    A field calibration method and results are described along with the experience gained with the method. The cup anemometers to be calibrated are mounted in a row on a 10-m high rig and calibrated in the free wind against a reference cup anemometer. The method has been reported [1] to improve...... the statistical bias on the data relative to calibrations carried out in a wind tunnel. The methodology is sufficiently accurate for calibration of cup anemometers used for wind resource assessments and provides a simple, reliable and cost-effective solution to cup anemometer calibration, especially suited...

  3. A quantum dot-aptamer beacon using a DNA intercalating dye as the FRET reporter: application to label-free thrombin detection.

    Science.gov (United States)

    Chi, Chun-Wei; Lao, Yeh-Hsing; Li, Yi-Shan; Chen, Lin-Chi

    2011-03-15

    A new quantum dot (QD)-aptamer (apt) beacon that acts by folding-induced dissociation of a DNA intercalating dye, BOBO-3(B), is demonstrated with label-free thrombin detection. The beacon, denoted as QD-apt:B, is constructed by (1) coupling of a single-stranded thrombin aptamer to Qdot 565 via EDC/Sulfo-NHS chemistry and (2) staining the duplex regions of the aptamer on QD with excess BOBO-3 before thrombin binding. When mixing a thrombin sample with QD-apt:B, BOBO-3 is competed away from the beacon due to target-induced aptamer folding, which then causes a decrease in QD fluorescence resonance energy transfer (FRET)-mediated BOBO-3 emission and achieves thrombin quantitation. In this work, the effects of Mg(2+), coupling time, and aptamer type on the beacon's performances are investigated and discussed thoroughly with various methods, including transmission electron microscopy (TEM), dynamic light scattering (DLS), and two-color differential gel electrophoresis. Using the best aptamer beacon (HTQ37), we attain highly specific and wide-range detection (from nM to μM) of thrombin in buffer, and the beacon can sense nM-range thrombin in 15% diluted serum. Compared to the reported QD aptamer assays, our method is advantageous from the aspect of using a simple sensory unit design without losing the detection sensitivity. Therefore, we consider the QD-apt:B beacon a potential alternative to immuno-reagents and an effective tool to study nucleic acid folding on QD as well. Copyright © 2011 Elsevier B.V. All rights reserved.

  4. Individual dosimetry and calibration

    International Nuclear Information System (INIS)

    Otto, T.

    1997-01-01

    In 1996, the Dosimetry and Calibration Section was, as in previous years, mainly engaged in routine tasks: the distribution of over 6000 dosimeters (with a total of more than 10,000 films) every two months and the calibration of about 900 fixed and mobile instruments used in the radiation survey sections of RP group. These tasks were, thanks to an experienced team, well mastered. Special efforts had to be made in a number of areas to modernize the service or to keep it in line with new prescriptions. The Individual Dosimetry Service had to assure that CERN's contracting firms comply with the prescriptions in the Radiation Safety Manual (1996) that had been inspired by the Swiss Ordinance of 1994: Companies must file for authorizations with the Swiss Federal Office for Public Health requiring that in every company an 'Expert in Radiation Protection' be nominated and subsequently trained. CERN's Individual Dosimetry Service is accredited by the Swiss Federal Authorities and works closely together with other, similar services on a rigorous quality assurance programme. Within this framework, CERN was mandated to organize this year the annual Swiss 'Intercomparison of Dosimeters'. All ten accredited dosimetry services - among others those of the Paul Scherrer Institute (PSI) in Villigen and of the four Swiss nuclear power stations - sent dosimeters to CERN, where they were irradiated in CERN's calibration facility with precise photon doses. After return to their origin they were processed and evaluated. The results were communicated to CERN and were compared with the originally given doses. A report on the results was subsequently prepared and submitted to the Swiss 'Group of Experts on Personal Dosimetry'. Reference monitors for photon and neutron radiation were brought to standard laboratories to assure the traceability of CERN's calibration service to the fundamental quantities. For photon radiation, a set of ionization chambers was calibrated in the reference field

  5. Sloan Digital Sky Survey photometric telescope automation and observing software

    International Nuclear Information System (INIS)

    Eric H. Neilsen, Jr.; email = neilsen@fnal.gov

    2002-01-01

    The photometric telescope (PT) provides observations necessary for the photometric calibration of the Sloan Digital Sky Survey (SDSS). Because the attention of the observing staff is occupied by the operation of the 2.5 meter telescope which takes the survey data proper, the PT must reliably take data with little supervision. In this paper we describe the PT's observing program, MOP, which automates most tasks necessary for observing. MOP's automated target selection is closely modeled on the actions a human observer might take, and is built upon a user interface that can be (and has been) used for manual operation. This results in an interface that makes it easy for an observer to track the activities of the automating procedures and intervene with minimum disturbance when necessary. MOP selects targets from the same list of standard star and calibration fields presented to the user, and chooses standard star fields covering ranges of airmass, color, and time necessary to monitor atmospheric extinction and produce a photometric solution. The software determines when additional standard star fields are unnecessary, and selects survey calibration fields according to availability and priority. Other automated features of MOP, such as maintaining the focus and keeping a night log, are also built around still functional manual interfaces, allowing the observer to be as active in observing as desired; MOP's automated features may be used as tools for manual observing, ignored entirely, or allowed to run the telescope with minimal supervision when taking routine data

  6. Automation in Clinical Microbiology

    Science.gov (United States)

    Ledeboer, Nathan A.

    2013-01-01

    Historically, the trend toward automation in clinical pathology laboratories has largely bypassed the clinical microbiology laboratory. In this article, we review the historical impediments to automation in the microbiology laboratory and offer insight into the reasons why we believe that we are on the cusp of a dramatic change that will sweep a wave of automation into clinical microbiology laboratories. We review the currently available specimen-processing instruments as well as the total laboratory automation solutions. Lastly, we outline the types of studies that will need to be performed to fully assess the benefits of automation in microbiology laboratories. PMID:23515547

  7. MAVEN SEP Calibrated Data Bundle

    Data.gov (United States)

    National Aeronautics and Space Administration — The maven.sep.calibrated Level 2 Science Data Bundle contains fully calibrated SEP data, as well as the raw count data from which they are derived, and ancillary...

  8. Ultrasonic calibration assembly

    International Nuclear Information System (INIS)

    1981-01-01

    Ultrasonic transducers for in-service inspection of nuclear reactor vessels have several problems associated with them which this invention seeks to overcome. The first is that of calibration or referencing a zero start point for the vertical axis of transducer movement to locate a weld defect. The second is that of verifying the positioning (vertically or at a predetermined angle). Thirdly there is the problem of ascertaining the speed per unit distance in the operating medium of the transducer beam prior to the actual inspection. The apparatus described is a calibration assembly which includes a fixed, generally spherical body having a surface for reflecting an ultrasonic beam from one of the transducers which can be moved until the reflection from the spherical body is the highest amplitude return signal indicating radial alignment from the body. (U.K.)

  9. Travelling gradient thermocouple calibration

    International Nuclear Information System (INIS)

    Broomfield, G.H.

    1975-01-01

    A short discussion of the origins of the thermocouple EMF is used to re-introduce the idea that the Peltier and Thompson effects are indistinguishable from one another. Thermocouples may be viewed as devices which generate an EMF at junctions or as integrators of EMF's developed in thermal gradients. The thermal gradient view is considered the more appropriate, because of its better accord with theory and behaviour, the correct approach to calibration, and investigation of service effects is immediately obvious. Inhomogeneities arise in thermocouples during manufacture and in service. The results of travelling gradient measurements are used to show that such effects are revealed with a resolution which depends on the length of the gradient although they may be masked during simple immersion calibration. Proposed tests on thermocouples irradiated in a nuclear reactor are discussed

  10. Mesoscale hybrid calibration artifact

    Science.gov (United States)

    Tran, Hy D.; Claudet, Andre A.; Oliver, Andrew D.

    2010-09-07

    A mesoscale calibration artifact, also called a hybrid artifact, suitable for hybrid dimensional measurement and the method for make the artifact. The hybrid artifact has structural characteristics that make it suitable for dimensional measurement in both vision-based systems and touch-probe-based systems. The hybrid artifact employs the intersection of bulk-micromachined planes to fabricate edges that are sharp to the nanometer level and intersecting planes with crystal-lattice-defined angles.

  11. Calibration of germanium detectors

    International Nuclear Information System (INIS)

    Bjurman, B.; Erlandsson, B.

    1985-01-01

    This paper describes problems concerning the calibration of germanium detectors for the measurement of gamma-radiation from environmental samples. It also contains a brief description of some ways of reducing the uncertainties concerning the activity determination. These uncertainties have many sources, such as counting statistics, full energy peak efficiency determination, density correction and radionuclide specific-coincidence effects, when environmental samples are investigated at close source-to-detector distances

  12. Calibration of hydrometers

    Science.gov (United States)

    Lorefice, Salvatore; Malengo, Andrea

    2006-10-01

    After a brief description of the different methods employed in periodic calibration of hydrometers used in most cases to measure the density of liquids in the range between 500 kg m-3 and 2000 kg m-3, particular emphasis is given to the multipoint procedure based on hydrostatic weighing, known as well as Cuckow's method. The features of the calibration apparatus and the procedure used at the INRiM (formerly IMGC-CNR) density laboratory have been considered to assess all relevant contributions involved in the calibration of different kinds of hydrometers. The uncertainty is strongly dependent on the kind of hydrometer; in particular, the results highlight the importance of the density of the reference buoyant liquid, the temperature of calibration and the skill of operator in the reading of the scale in the whole assessment of the uncertainty. It is also interesting to realize that for high-resolution hydrometers (division of 0.1 kg m-3), the uncertainty contribution of the density of the reference liquid is the main source of the total uncertainty, but its importance falls under about 50% for hydrometers with a division of 0.5 kg m-3 and becomes somewhat negligible for hydrometers with a division of 1 kg m-3, for which the reading uncertainty is the predominant part of the total uncertainty. At present the best INRiM result is obtained with commercially available hydrometers having a scale division of 0.1 kg m-3, for which the relative uncertainty is about 12 × 10-6.

  13. Observation models in radiocarbon calibration

    International Nuclear Information System (INIS)

    Jones, M.D.; Nicholls, G.K.

    2001-01-01

    The observation model underlying any calibration process dictates the precise mathematical details of the calibration calculations. Accordingly it is important that an appropriate observation model is used. Here this is illustrated with reference to the use of reservoir offsets where the standard calibration approach is based on a different model to that which the practitioners clearly believe is being applied. This sort of error can give rise to significantly erroneous calibration results. (author). 12 refs., 1 fig

  14. Dosimetry and Calibration Section

    International Nuclear Information System (INIS)

    Otto, T.

    1998-01-01

    The two tasks of the Dosimetry and Calibration Section at CERN are the Individual Dosimetry Service which assures the personal monitoring of about 5000 persons potentially exposed to ionizing radiation at CERN, and the Calibration Laboratory which verifies all the instruments and monitors. This equipment is used by the sections of the RP Group for assuring radiation protection around CERN's accelerators, and by the Environmental Section of TISTE. In addition, nearly 250 electronic and 300 quartz fibre dosimeters, employed in operational dosimetry, are calibrated at least once a year. The Individual Dosimetry Service uses an extended database (INDOS) which contains information about all the individual doses ever received at CERN. For most of 1997 it was operated without the support of a database administrator as the technician who had assured this work retired. The Software Support Section of TIS-TE took over the technical responsibility of the database, but in view of the many other tasks of this Section and the lack of personnel, only a few interventions for solving immediate problems were possible

  15. Calibrated Properties Model

    International Nuclear Information System (INIS)

    Ghezzehej, T.

    2004-01-01

    The purpose of this model report is to document the calibrated properties model that provides calibrated property sets for unsaturated zone (UZ) flow and transport process models (UZ models). The calibration of the property sets is performed through inverse modeling. This work followed, and was planned in, ''Technical Work Plan (TWP) for: Unsaturated Zone Flow Analysis and Model Report Integration'' (BSC 2004 [DIRS 169654], Sections 1.2.6 and 2.1.1.6). Direct inputs to this model report were derived from the following upstream analysis and model reports: ''Analysis of Hydrologic Properties Data'' (BSC 2004 [DIRS 170038]); ''Development of Numerical Grids for UZ Flow and Transport Modeling'' (BSC 2004 [DIRS 169855]); ''Simulation of Net Infiltration for Present-Day and Potential Future Climates'' (BSC 2004 [DIRS 170007]); ''Geologic Framework Model'' (GFM2000) (BSC 2004 [DIRS 170029]). Additionally, this model report incorporates errata of the previous version and closure of the Key Technical Issue agreement TSPAI 3.26 (Section 6.2.2 and Appendix B), and it is revised for improved transparency

  16. Calibration of robotic drilling systems with a moving rail

    Directory of Open Access Journals (Sweden)

    Tian Wei

    2014-12-01

    Full Text Available Industrial robots are widely used in aircraft assembly systems such as robotic drilling systems. It is necessary to expand a robot’s working range with a moving rail. A method for improving the position accuracy of an automated assembly system with an industrial robot mounted on a moving rail is proposed. A multi-station method is used to control the robot in this study. The robot only works at stations which are certain positions defined on the moving rail. The calibration of the robot system is composed by the calibration of the robot and the calibration of the stations. The calibration of the robot is based on error similarity and inverse distance weighted interpolation. The calibration of the stations is based on a magnetic strip and a magnetic sensor. Validation tests were performed in this study, which showed that the accuracy of the robot system gained significant improvement using the proposed method. The absolute position errors were reduced by about 85% to less than 0.3 mm compared with the maximum nearly 2 mm before calibration.

  17. Thrombin-induced, TNFR-dependent miR-181c downregulation promotes MLL1 and NF-κB target gene expression in human microglia.

    Science.gov (United States)

    Yin, Min; Chen, Zhiying; Ouyang, Yetong; Zhang, Huiyan; Wan, Zhigang; Wang, Han; Wu, Wei; Yin, Xiaoping

    2017-06-29

    Controlling thrombin-driven microglial activation may serve as a therapeutic target for intracerebral hemorrhage (ICH). Here, we investigated microRNA (miRNA)-based regulation of thrombin-driven microglial activation using an in vitro thrombin toxicity model applied to primary human microglia. A miRNA array identified 22 differential miRNA candidates. Quantitative real-time reverse transcription polymerase chain reaction (qRT-PCR) identified miR-181c as the most significantly downregulated miRNA. TargetScan analysis identified mixed lineage leukemia-1 (MLL1) as a putative gene target for miR-181c. qRT-PCR was applied to assess tumor necrosis factor-alpha (TNF-α), miR-181c, and MLL1 levels following thrombin or proteinase-activated receptor-4-specific activating peptide (PAR4AP) exposure. Anti-TNF-α antibodies and tumor necrosis factor receptor (TNFR) silencing were employed to test TNF-α/TNFR dependence. A dual-luciferase reporter system and miR-181c mimic transfection assessed whether mir-181c directly binds to and negatively regulates MLL1. Nuclear factor kappa-B (NF-κB)-dependent luciferase reporter assays and NF-κB target gene expression were assessed in wild-type (MLL1+) and MLL1-silenced cells. Thrombin or PAR4AP-induced miR-181c downregulation (p < 0.05) and MLL1 upregulation (p < 0.05) that were dependent upon TNF-α/TNFR. miR-181c decreased wild-type MLL1 3'-UTR luciferase reporter activity (p < 0.05), and a miR-181c mimic suppressed MLL1 expression (p < 0.05). Thrombin treatment increased, while miR-181c reduced, NF-κB activity and NF-κB target gene expression in both wild-type (MLL1+) and MLL1-silenced cells (p < 0.05). Thrombin-induced, TNF-α/TNFR-dependent miR-181c downregulation promotes MLL1 expression, increases NF-κB activity, and upregulates NF-κB target gene expression. As miR-181c opposes thrombin's stimulation of pro-inflammatory NF-κB activity, miR-181c mimic therapy may show promise in controlling thrombin

  18. Phage display of the serpin alpha-1 proteinase inhibitor randomized at consecutive residues in the reactive centre loop and biopanned with or without thrombin.

    Directory of Open Access Journals (Sweden)

    Benjamin M Scott

    Full Text Available In spite of the power of phage display technology to identify variant proteins with novel properties in large libraries, it has only been previously applied to one member of the serpin superfamily. Here we describe phage display of human alpha-1 proteinase inhibitor (API in a T7 bacteriophage system. API M358R fused to the C-terminus of T7 capsid protein 10B was directly shown to form denaturation-resistant complexes with thrombin by electrophoresis and immunoblotting following exposure of intact phages to thrombin. We therefore developed a biopanning protocol in which thrombin-reactive phages were selected using biotinylated anti-thrombin antibodies and streptavidin-coated magnetic beads. A library consisting of displayed API randomized at residues 357 and 358 (P2-P1 yielded predominantly Pro-Arg at these positions after five rounds of thrombin selection; in contrast the same degree of mock selection yielded only non-functional variants. A more diverse library of API M358R randomized at residues 352-356 (P7-P3 was also probed, yielding numerous variants fitting a loose consensus of DLTVS as judged by sequencing of the inserts of plaque-purified phages. The thrombin-selected sequences were transferred en masse into bacterial expression plasmids, and lysates from individual colonies were screening for API-thrombin complexing. The most active candidates from this sixth round of screening contained DITMA and AAFVS at P7-P3 and inhibited thrombin 2.1-fold more rapidly than API M358R with no change in reaction stoichiometry. Deep sequencing using the Ion Torrent platform confirmed that over 800 sequences were significantly enriched in the thrombin-panned versus naïve phage display library, including some detected using the combined phage display/bacterial lysate screening approach. Our results show that API joins Plasminogen Activator Inhibitor-1 (PAI-1 as a serpin amenable to phage display and suggest the utility of this approach for the selection

  19. Enhanced Effector Function of CD8+ T Cells From Healthy Controls and HIV-Infected Patients Occurs Through Thrombin Activation of Protease-Activated Receptor 1

    Science.gov (United States)

    Hurley, Amanda; Smith, Mindy; Karpova, Tatiana; Hasley, Rebecca B.; Belkina, Natalya; Shaw, Stephen; Balenga, Nariman; Druey, Kirk M.; Nickel, Erin; Packard, Beverly; Imamichi, Hiromi; Hu, Zonghui; Follmann, Dean; McNally, James; Higgins, Jeanette; Sneller, Michael; Lane, H. Clifford; Catalfamo, Marta

    2013-01-01

    Disruption of vascular integrity by trauma and other tissue insults leads to inflammation and activation of the coagulation cascade. The serine protease thrombin links these 2 processes. The proinflammatory function of thrombin is mediated by activation of protease-activated receptor 1 (PAR-1). We found that peripheral blood effector memory CD4+ and CD8+ T lymphocytes expressed PAR-1 and that expression was increased in CD8+ T cells from human immunodeficiency virus (HIV)–infected patients. Thrombin enhanced cytokine secretion in CD8+ T cells from healthy controls and HIV-infected patients. In addition, thrombin induced chemokinesis, but not chemotaxis, of CD8+ T cells, which led to structural changes, including cell polarization and formation of a structure rich in F-actin and phosphorylated ezrin-radexin-moesin proteins. These findings suggest that thrombin mediates cross-talk between the coagulation system and the adaptive immune system at sites of vascular injury through increased T-cell motility and production of proinflammatory cytokines. PMID:23204166

  20. Enhanced effector function of CD8(+) T cells from healthy controls and HIV-infected patients occurs through thrombin activation of protease-activated receptor 1.

    Science.gov (United States)

    Hurley, Amanda; Smith, Mindy; Karpova, Tatiana; Hasley, Rebecca B; Belkina, Natalya; Shaw, Stephen; Balenga, Nariman; Druey, Kirk M; Nickel, Erin; Packard, Beverly; Imamichi, Hiromi; Hu, Zonghui; Follmann, Dean; McNally, James; Higgins, Jeanette; Sneller, Michael; Lane, H Clifford; Catalfamo, Marta

    2013-02-15

    Disruption of vascular integrity by trauma and other tissue insults leads to inflammation and activation of the coagulation cascade. The serine protease thrombin links these 2 processes. The proinflammatory function of thrombin is mediated by activation of protease-activated receptor 1 (PAR-1). We found that peripheral blood effector memory CD4(+) and CD8(+) T lymphocytes expressed PAR-1 and that expression was increased in CD8(+) T cells from human immunodeficiency virus (HIV)-infected patients. Thrombin enhanced cytokine secretion in CD8(+) T cells from healthy controls and HIV-infected patients. In addition, thrombin induced chemokinesis, but not chemotaxis, of CD8(+) T cells, which led to structural changes, including cell polarization and formation of a structure rich in F-actin and phosphorylated ezrin-radexin-moesin proteins. These findings suggest that thrombin mediates cross-talk between the coagulation system and the adaptive immune system at sites of vascular injury through increased T-cell motility and production of proinflammatory cytokines.

  1. A mathematical model for camera calibration based on straight lines

    Directory of Open Access Journals (Sweden)

    Antonio M. G. Tommaselli

    2005-12-01

    Full Text Available In other to facilitate the automation of camera calibration process, a mathematical model using straight lines was developed, which is based on the equivalent planes mathematical model. Parameter estimation of the developed model is achieved by the Least Squares Method with Conditions and Observations. The same method of adjustment was used to implement camera calibration with bundles, which is based on points. Experiments using simulated and real data have shown that the developed model based on straight lines gives results comparable to the conventional method with points. Details concerning the mathematical development of the model and experiments with simulated and real data will be presented and the results with both methods of camera calibration, with straight lines and with points, will be compared.

  2. Calibration technology in application of robot-laser scanning system

    Science.gov (United States)

    Ren, YongJie; Yin, ShiBin; Zhu, JiGui

    2012-11-01

    A system composed of laser sensor and 6-DOF industrial robot is proposed to obtain complete three-dimensional (3-D) information of the object surface. Suitable for the different combining ways of laser sensor and robot, a new method to calibrate the position and pose between sensor and robot is presented. By using a standard sphere with known radius as a reference tool, the rotation and translation matrices between the laser sensor and robot are computed, respectively in two steps, so that many unstable factors introduced in conventional optimization methods can be avoided. The experimental results show that the accuracy of the proposed calibration method can be achieved up to 0.062 mm. The calibration method is also implemented into the automated robot scanning system to reconstruct a car door panel.

  3. Calibration methods for ECE systems with microwave sources

    International Nuclear Information System (INIS)

    Tubbing, B.J.D.; Kissel, S.E.

    1987-01-01

    The authors investigated the feasibility of two methods for calibration of electron cyclotron emission (ECE) systems, both based on the use of a microwave source. In the first method -called the Antenna Pattern Integration (API) method - the microwave source is scanned in space, so as to simulate a large - area - blackbody -source. In the second method -called the Untuned Cavity (UC) method -an untuned cavity, fed by the microwave source, is used to simulate a blackbody. For both methods, the hardware required to perform partly automated calibrations was developed. The microwave based methods were compared with a large area blackbody calibration on two different ECE systems, a Michelson interferometer and a grating polychromator. The API method was found to be more successful than the UC method. (author)

  4. Pilot study to verify the calibration of electrometers

    International Nuclear Information System (INIS)

    Becker, P.; Meghzifene, A.

    2002-01-01

    Full text: The main detector used for standardization of the quantities used in measurements of ionizing radiation is the ionization chamber. The interaction of the radiation with this detector produces electrical charge, usually, in the range of pC to nC. The instrument used to measure such small charges (or currents) is the electrometer. As part of a good practice, the measured charge (current) must be traceable to a primary or secondary standard. Some calibration laboratories can only provide a system calibration coefficient, i.e. a calibration coefficient for the combination of electrometer plus ionization chamber (Gy/scale division). This practice is acceptable, but it can impose a limitation to the automation of their calibration procedures (using computerized application for the acquisition of current/charge). Not all models have the possibility of a connection to a computer and in the case of those that don't have this capability, automation is not possible without the development of a specific interface. In addition, end-users receive a calibration coefficient, which is only valid for the set ion chamber and electrometer. In case of a broken chamber, the end-user cannot connect another chamber to their electrometer without knowing its calibration coefficient. If the calibration laboratories had the capability of calibrating the chamber separately from the electrometer, for example, using an electrometer calibrated in terms of charge, all the chambers could be calibrated using this electrometer. The laboratory can also benefit from the automation of the measurements. This requires that the laboratory must be able to cross-calibrate the electrometers (associated to the chambers) also in terms of charge (Coulombs). Electrical charge is standardized by the use of a standard air capacitor and a standard voltage source (Q=CV) and the National Laboratory for Metrology of Ionizing Radiation (IRD) in Brazil has also adopted this procedure. Since the Brazilian

  5. Thrombin-induced rabbit platelet microbicidal protein is fungicidal in vitro.

    Science.gov (United States)

    Yeaman, M R; Ibrahim, A S; Edwards, J E; Bayer, A S; Ghannoum, M A

    1993-03-01

    Platelet microbicidal protein (PMP) is released from platelets in response to thrombin stimulation. PMP is known to possess in vitro bactericidal activity against Staphylococcus aureus and viridans group streptococci. To determine whether PMP is active against other intravascular pathogens, we evaluated its potential fungicidal activity against strains of Candida species and Cryptococcus neoformans. Anionic resin adsorption and gel electrophoresis confirmed that the fungicidal activity of PMP resided in a small (approximately 8.5-kDa), cationic protein, identical to previous studies of PMP-induced bacterial killing (M.R. Yeaman, S.M. Puentes, D.C. Norman, and A.S. Bayer, Infect. Immun. 60:1202-1209, 1992). When assayed over a 180-min period in vitro, the susceptibilities of these fungi to PMP varied considerably. Generally, Candida albicans strains (mean survival, 33.5% +/- 6.9% [n = 6]) as well as isolates of Candida glabrata (mean survival, 50.8% +/- 2.9% [n = 2]) were the most susceptible to killing by PMP, while Candida guillermondii and Candida parapsilosis were relatively resistant to PMP-induced killing. Compared with C. albicans, C. neoformans was relatively resistant to the fungicidal activity of PMP, with a mean survival among the isolates studied of 77.4% +/- 12.4% (n = 6). Against C. albicans, PMP-induced fungicidal activity was time dependent (range, 0 to 180 min), PMP concentration dependent (range, 10 to 150 U/ml), and inversely related to the fungal inoculum (range, 5 x 10(3) to 1 x 10(5) CFU/ml). Scanning electron microscopy of PMP-exposed C. albicans and C. neoformans cells revealed extensive surface damage and collapse, suggesting that the site of PMP fungicidal action may directly or indirectly involve the fungal cell envelope.

  6. Sphingosine kinase inhibition alleviates endothelial permeability induced by thrombin and activated neutrophils.

    Science.gov (United States)

    Itagaki, Kiyoshi; Zhang, Qin; Hauser, Carl J

    2010-04-01

    Inflammation and microvascular thrombosis are interrelated causes of acute lung injury in the systemic inflammatory response syndrome. Neutrophils (polymorphonuclear neutrophil [PMN]) and endothelial cells (EC) activated by systemic inflammatory response syndrome interact to increase pulmonary vascular permeability, but the interactions between PMN and EC are difficult to study. Recently, we reported that sphingosine 1-phosphate is a second messenger eliciting store-operated calcium entry (SOCE) in response to inflammatory agonists in both PMN and EC. Store-operated calcium entry is therefore a target mechanism for the therapeutic modulation of inflammatory PMN-EC interactions. Here, we isolated, modeled, and studied the effects of pharmacologic SOCE inhibition using real-time systems to monitor EC permeability after exposure to activated PMN. We created systems to continuously assess permeability of human pulmonary artery endothelial cells and human microvascular endothelial cells from lung. Endothelial cells show increased permeability after challenge by activated PMN. Such permeability increases can be attenuated by exposure of the cocultures to sphingosine kinase (SK) inhibitors (SKI-2, N,N-dimethylsphingosine [DMS]) or Ca2+ entry inhibitors (Gd3+, MRS-1845). Human microvascular endothelial cells from lung pretreated with SKI-2 or DMS showed decreased permeability when later exposed to activated PMN. Likewise, when PMNs were activated with thapsigargin (TG) in the presence of SKI-2, DMS, Gd, or MRS-1845, their ability to cause EC permeability subsequently was reduced. SKI-2 also inhibited the activation of human pulmonary artery ECs by thrombin. These studies will provide a firm mechanistic foundation for understanding how systemic SOCE inhibition may be used to prevent acute lung injury in vivo.

  7. Assessment of thrombus imaging potency of thrombin-targeting recombinant hirudin in vitro and in vivo

    Energy Technology Data Exchange (ETDEWEB)

    Cheng, Tan; Rongjun, Zhang; Weixing, Wan; Gangming, Cai; Huixin, Yu; Mengjun, Jiang; Lianfen, Zhang; Guoping, Sheng; Yijun, Fan; Yonghui, Tao; Jian, Jin [Jiangsu Inst. of Nuclear Medicine, Wuxi (China)

    2003-07-01

    The purpose of this study is to evaluate the effect of recombinant hirudin HV2 (rHHV2) as a thrombus imaging agent. {sup 125}I-rHHV2 and {sup 125}I-Th were prepared with Chloramine method, the labeling rate were 86.64% and 62.20%, with the radioactive purity of 89.70% and 91.22%, with the specific activity of 22.4 TBq/mmol and 94.43 TBq/mmol respectively. The competitive radioassay showed that the Th-fibrin complex formation did not affect the ability of rHHV2 binding with Th. In the complex, the molecular binding ratio of rHHV2 to Th and fibrinogen was 14:14:1. {sup 99m}Tc-rHHV2 was prepared by 2-iminothiolane modified method, the labeled rate was 94%, with the radioactive purity of 93.90%, with the specific activity of 2.30 TBq/mmol. It was used to image fresh thrombi on arteries and veins of dog or rabbit (30 {mu}g/kg). In SPECT images, all thrombin were clearly visible, arterial thrombosis imaging can be seen clearly within 45 min after injection and fade away slowly, venous thrombosis imaging also can be seen within 30 min after injection and quantitative imaging ratios between the thrombus and opposite vessel increased following the time. Biodistribution studies in mouse demonstrated that rHHV2 was excreted from kidneys. These data indicate that Th in Th-fibrin complex could be a potent target for diagnosis of thrombus and {sup 99m}Tc-rHHV2 could be a new thrombotic imaging agent. (authors)

  8. Building a framework to manage trust in automation

    Science.gov (United States)

    Metcalfe, J. S.; Marathe, A. R.; Haynes, B.; Paul, V. J.; Gremillion, G. M.; Drnec, K.; Atwater, C.; Estepp, J. R.; Lukos, J. R.; Carter, E. C.; Nothwang, W. D.

    2017-05-01

    All automations must, at some point in their lifecycle, interface with one or more humans. Whether operators, end-users, or bystanders, human responses can determine the perceived utility and acceptance of an automation. It has been long believed that human trust is a primary determinant of human-automation interactions and further presumed that calibrating trust can lead to appropriate choices regarding automation use. However, attempts to improve joint system performance by calibrating trust have not yet provided a generalizable solution. To address this, we identified several factors limiting the direct integration of trust, or metrics thereof, into an active mitigation strategy. The present paper outlines our approach to addressing this important issue, its conceptual underpinnings, and practical challenges encountered in execution. Among the most critical outcomes has been a shift in focus from trust to basic interaction behaviors and their antecedent decisions. This change in focus inspired the development of a testbed and paradigm that was deployed in two experiments of human interactions with driving automation that were executed in an immersive, full-motion simulation environment. Moreover, by integrating a behavior and physiology-based predictor within a novel consequence-based control system, we demonstrated that it is possible to anticipate particular interaction behaviors and influence humans towards more optimal choices about automation use in real time. Importantly, this research provides a fertile foundation for the development and integration of advanced, wearable technologies for sensing and inferring critical state variables for better integration of human elements into otherwise fully autonomous systems.

  9. Comprehensive Calibration and Validation Site for Information Remote Sensing

    Science.gov (United States)

    Li, C. R.; Tang, L. L.; Ma, L. L.; Zhou, Y. S.; Gao, C. X.; Wang, N.; Li, X. H.; Wang, X. H.; Zhu, X. H.

    2015-04-01

    As a naturally part of information technology, Remote Sensing (RS) is strongly required to provide very precise and accurate information product to serve industry, academy and the public at this information economic era. To meet the needs of high quality RS product, building a fully functional and advanced calibration system, including measuring instruments, measuring approaches and target site become extremely important. Supported by MOST of China via national plan, great progress has been made to construct a comprehensive calibration and validation (Cal&Val) site, which integrates most functions of RS sensor aviation testing, EO satellite on-orbit caration and performance assessment and RS product validation at this site located in Baotou, 600km west of Beijing. The site is equipped with various artificial standard targets, including portable and permanent targets, which supports for long-term calibration and validation. A number of fine-designed ground measuring instruments and airborne standard sensors are developed for realizing high-accuracy stepwise validation, an approach in avoiding or reducing uncertainties caused from nonsynchronized measurement. As part of contribution to worldwide Cal&Val study coordinated by CEOS-WGCV, Baotou site is offering its support to Radiometric Calibration Network of Automated Instruments (RadCalNet), with an aim of providing demonstrated global standard automated radiometric calibration service in cooperation with ESA, NASA, CNES and NPL. Furthermore, several Cal&Val campaigns have been performed during the past years to calibrate and validate the spaceborne/airborne optical and SAR sensors, and the results of some typical demonstration are discussed in this study.

  10. Calibration of triaxial fluxgate gradiometer

    International Nuclear Information System (INIS)

    Vcelak, Jan

    2006-01-01

    The description of simple and fast calibration procedures used for double-probe triaxial fluxgate gradiometer is provided in this paper. The calibration procedure consists of three basic steps. In the first step both probes are calibrated independently in order to reach constant total field reading in every position. Both probes are numerically aligned in the second step in order that the gradient reading is zero in homogenous magnetic field. The third step consists of periodic drift calibration during measurement. The results and detailed description of each calibration step are presented and discussed in the paper. The gradiometer is finally verified during the detection of the metal object in the measuring grid

  11. Primary calibration in acoustics metrology

    International Nuclear Information System (INIS)

    Milhomem, T A Bacelar; Soares, Z M Defilippo

    2015-01-01

    SI unit in acoustics is realized by the reciprocity calibrations of laboratory standard microphones in pressure field, free field and diffuse field. Calibrations in pressure field and in free field are already consolidated and the Inmetro already done them. Calibration in diffuse field is not yet consolidated, however, some national metrology institutes, including Inmetro, are conducting researches on this subject. This paper presents the reciprocity calibration, the results of Inmetro in recent key comparisons and the research that is being developed for the implementation of reciprocity calibration in diffuse field

  12. Mercury CEM Calibration

    Energy Technology Data Exchange (ETDEWEB)

    John Schabron; Joseph Rovani; Mark Sanderson

    2008-02-29

    Mercury continuous emissions monitoring systems (CEMS) are being implemented in over 800 coal-fired power plant stacks. The power industry desires to conduct at least a full year of monitoring before the formal monitoring and reporting requirement begins on January 1, 2009. It is important for the industry to have available reliable, turnkey equipment from CEM vendors. Western Research Institute (WRI) is working closely with the Electric Power Research Institute (EPRI), the National Institute of Standards and Technology (NIST), and the Environmental Protection Agency (EPA) to facilitate the development of the experimental criteria for a NIST traceability protocol for dynamic elemental mercury vapor generators. The generators are used to calibrate mercury CEMs at power plant sites. The Clean Air Mercury Rule (CAMR) which was published in the Federal Register on May 18, 2005 requires that calibration be performed with NIST-traceable standards (Federal Register 2007). Traceability procedures will be defined by EPA. An initial draft traceability protocol was issued by EPA in May 2007 for comment. In August 2007, EPA issued an interim traceability protocol for elemental mercury generators (EPA 2007). The protocol is based on the actual analysis of the output of each calibration unit at several concentration levels ranging initially from about 2-40 {micro}g/m{sup 3} elemental mercury, and in the future down to 0.2 {micro}g/m{sup 3}, and this analysis will be directly traceable to analyses by NIST. The document is divided into two separate sections. The first deals with the qualification of generators by the vendors for use in mercury CEM calibration. The second describes the procedure that the vendors must use to certify the generator models that meet the qualification specifications. The NIST traceable certification is performance based, traceable to analysis using isotope dilution inductively coupled plasma/mass spectrometry performed by NIST in Gaithersburg, MD. The

  13. Roles of Platelet STIM1 and Orai1 in Glycoprotein VI- and Thrombin-dependent Procoagulant Activity and Thrombus Formation*

    Science.gov (United States)

    Gilio, Karen; van Kruchten, Roger; Braun, Attila; Berna-Erro, Alejandro; Feijge, Marion A. H.; Stegner, David; van der Meijden, Paola E. J.; Kuijpers, Marijke J. E.; Varga-Szabo, David; Heemskerk, Johan W. M.; Nieswandt, Bernhard

    2010-01-01

    In platelets, STIM1 has been recognized as the key regulatory protein in store-operated Ca2+ entry (SOCE) with Orai1 as principal Ca2+ entry channel. Both proteins contribute to collagen-dependent arterial thrombosis in mice in vivo. It is unclear whether STIM2 is involved. A key platelet response relying on Ca2+ entry is the surface exposure of phosphatidylserine (PS), which accomplishes platelet procoagulant activity. We studied this response in mouse platelets deficient in STIM1, STIM2, or Orai1. Upon high shear flow of blood over collagen, Stim1−/− and Orai1−/− platelets had greatly impaired glycoprotein (GP) VI-dependent Ca2+ signals, and they were deficient in PS exposure and thrombus formation. In contrast, Stim2−/− platelets reacted normally. Upon blood flow in the presence of thrombin generation and coagulation, Ca2+ signals of Stim1−/− and Orai1−/− platelets were partly reduced, whereas the PS exposure and formation of fibrin-rich thrombi were normalized. Washed Stim1−/− and Orai1−/− platelets were deficient in GPVI-induced PS exposure and prothrombinase activity, but not when thrombin was present as co-agonist. Markedly, SKF96365, a blocker of (receptor-operated) Ca2+ entry, inhibited Ca2+ and procoagulant responses even in Stim1−/− and Orai1−/− platelets. These data show for the first time that: (i) STIM1 and Orai1 jointly contribute to GPVI-induced SOCE, procoagulant activity, and thrombus formation; (ii) a compensating Ca2+ entry pathway is effective in the additional presence of thrombin; (iii) platelets contain two mechanisms of Ca2+ entry and PS exposure, only one relying on STIM1-Orai1 interaction. PMID:20519511

  14. Roles of platelet STIM1 and Orai1 in glycoprotein VI- and thrombin-dependent procoagulant activity and thrombus formation.

    Science.gov (United States)

    Gilio, Karen; van Kruchten, Roger; Braun, Attila; Berna-Erro, Alejandro; Feijge, Marion A H; Stegner, David; van der Meijden, Paola E J; Kuijpers, Marijke J E; Varga-Szabo, David; Heemskerk, Johan W M; Nieswandt, Bernhard

    2010-07-30

    In platelets, STIM1 has been recognized as the key regulatory protein in store-operated Ca(2+) entry (SOCE) with Orai1 as principal Ca(2+) entry channel. Both proteins contribute to collagen-dependent arterial thrombosis in mice in vivo. It is unclear whether STIM2 is involved. A key platelet response relying on Ca(2+) entry is the surface exposure of phosphatidylserine (PS), which accomplishes platelet procoagulant activity. We studied this response in mouse platelets deficient in STIM1, STIM2, or Orai1. Upon high shear flow of blood over collagen, Stim1(-/-) and Orai1(-/-) platelets had greatly impaired glycoprotein (GP) VI-dependent Ca(2+) signals, and they were deficient in PS exposure and thrombus formation. In contrast, Stim2(-/-) platelets reacted normally. Upon blood flow in the presence of thrombin generation and coagulation, Ca(2+) signals of Stim1(-/-) and Orai1(-/-) platelets were partly reduced, whereas the PS exposure and formation of fibrin-rich thrombi were normalized. Washed Stim1(-/-) and Orai1(-/-) platelets were deficient in GPVI-induced PS exposure and prothrombinase activity, but not when thrombin was present as co-agonist. Markedly, SKF96365, a blocker of (receptor-operated) Ca(2+) entry, inhibited Ca(2+) and procoagulant responses even in Stim1(-/-) and Orai1(-/-) platelets. These data show for the first time that: (i) STIM1 and Orai1 jointly contribute to GPVI-induced SOCE, procoagulant activity, and thrombus formation; (ii) a compensating Ca(2+) entry pathway is effective in the additional presence of thrombin; (iii) platelets contain two mechanisms of Ca(2+) entry and PS exposure, only one relying on STIM1-Orai1 interaction.

  15. In vitro assessment of Tc-99m labeled bovine thrombin and streptokinase-activated human plasmin: concise communication. [Iodine 125

    Energy Technology Data Exchange (ETDEWEB)

    Wong, D.W.; Tanaka, T.; Mishkin, F.; Lee, T.

    1979-09-01

    Bovine thrombin and streptokinase-activated human plasmin have been labeled with Tc-99m using stannous reduction of pertechnetate under physiological conditions (pH 7.4). The binding efficiency of radiotechnetium to these enzymes is greater than 94%, with less than 5% of reduced but unbound Tc-99m (Sn) complex as assayed by ascending paper radiochromatography using ITLC silica gel plate. Free or unbound pertechnetate is less than 1%. In vitro enzymatic analyses of the Tc-99m-labeled enzymes demonstrate no evidence of protein denaturation or significant loss of enzymatic activity after labeling. Both labeled enzymes are biochemically active in vitro with their respective substrates.

  16. In vitro assessment of Tc-99m labeled bovine thrombin and streptokinase-activated human plasmin: concise communication

    International Nuclear Information System (INIS)

    Wong, D.W.; Tanaka, T.; Mishkin, F.; Lee, T.

    1979-01-01

    Bovine thrombin and streptokinase-activated human plasmin have been labeled with Tc-99m using stannous reduction of pertechnetate under physiological conditions (pH 7.4). The binding efficiency of radiotechnetium to these enzymes is greater than 94%, with less than 5% of reduced but unbound Tc-99m (Sn) complex as assayed by ascending paper radiochromatography using ITLC silica gel plate. Free or unbound pertechnetate is less than 1%. In vitro enzymatic analyses of the Tc-99m-labeled enzymes demonstrate no evidence of protein denaturation or significant loss of enzymatic activity after labeling. Both labeled enzymes are biochemically active in vitro with their respective substrates

  17. Host defense peptides of thrombin modulate inflammation and coagulation in endotoxin-mediated shock and Pseudomonas aeruginosa sepsis

    DEFF Research Database (Denmark)

    Kalle, Martina; Papareddy, Praveen; Kasetty, Gopinath

    2012-01-01

    Gram-negative sepsis is accompanied by a disproportionate innate immune response and excessive coagulation mainly induced by endotoxins released from bacteria. Due to rising antibiotic resistance and current lack of other effective treatments there is an urgent need for new therapies. We here...... present a new treatment concept for sepsis and endotoxin-mediated shock, based on host defense peptides from the C-terminal part of human thrombin, found to have a broad and inhibitory effect on multiple sepsis pathologies. Thus, the peptides abrogate pro-inflammatory cytokine responses to endotoxin...

  18. The CHEOPS calibration bench

    Science.gov (United States)

    Wildi, F.; Chazelas, B.; Deline, A.; Sarajlic, M.; Sordet, M.

    2017-09-01

    CHEOPS is an ESA Class S Mission aiming at the characterization of exoplanets through the precise measurement of their radius, using the transit method [1]. To achieve this goal, the payload is designed to be a high precision "absolute" photometer, looking at one star at a time. It will be able to cover la large fraction of the sky by repointing. Its launch is expected at the end of 2017 [2, this conference]. CHEOPS' main science is the measure of the transit of exoplanets of radius ranging from 1 to 6 Earth radii orbiting bright stars. The required photometric stability to reach this goal is of 20 ppm in 6 hours for a 9th magnitude star. The CHEOPS' only instrument is a Ritchey-Chretien style telescope with 300 mm effective aperture diameter, which provides a defocussed image of the target star on a single frame-transfer backside illuminated CCD detector cooled to -40°C and stabilized within 10 mK [2]. CHEOPS being in a LEO, it is equipped with a high performance baffle. The spacecraft platform provides a pointing stability of < 2 arcsec rms. This relatively modest pointing performance makes high quality flat-fielding necessary In the rest of this article we will refer to the only CHEOPS instrument simply as "CHEOP" Its behavior will be calibrated thoroughly on the ground and only a small subset of the calibrations can be redone in flight. The main focuses of the calibrations are the photonic gain stability and sensibility to the environment variations and the Flat field that has to be known at a precision better than 0.1%.

  19. CALIBRATED HYDRODYNAMIC MODEL

    Directory of Open Access Journals (Sweden)

    Sezar Gülbaz

    2015-01-01

    Full Text Available The land development and increase in urbanization in a watershed affect water quantityand water quality. On one hand, urbanization provokes the adjustment of geomorphicstructure of the streams, ultimately raises peak flow rate which causes flood; on theother hand, it diminishes water quality which results in an increase in Total SuspendedSolid (TSS. Consequently, sediment accumulation in downstream of urban areas isobserved which is not preferred for longer life of dams. In order to overcome thesediment accumulation problem in dams, the amount of TSS in streams and inwatersheds should be taken under control. Low Impact Development (LID is a BestManagement Practice (BMP which may be used for this purpose. It is a land planningand engineering design method which is applied in managing storm water runoff inorder to reduce flooding as well as simultaneously improve water quality. LID includestechniques to predict suspended solid loads in surface runoff generated over imperviousurban surfaces. In this study, the impact of LID-BMPs on surface runoff and TSS isinvestigated by employing a calibrated hydrodynamic model for Sazlidere Watershedwhich is located in Istanbul, Turkey. For this purpose, a calibrated hydrodynamicmodel was developed by using Environmental Protection Agency Storm WaterManagement Model (EPA SWMM. For model calibration and validation, we set up arain gauge and a flow meter into the field and obtain rainfall and flow rate data. Andthen, we select several LID types such as retention basins, vegetative swales andpermeable pavement and we obtain their influence on peak flow rate and pollutantbuildup and washoff for TSS. Consequently, we observe the possible effects ofLID on surface runoff and TSS in Sazlidere Watershed.

  20. Radionuclide calibrators performance evaluation

    International Nuclear Information System (INIS)

    Mora Ramirez, E.; Zeledon Fonseca, P.; Jimenez Cordero, M.

    2008-01-01

    Radionuclide calibrators are used to estimate accurately activity prior to administration to a patient, so it is very important that this equipment meets its performance requirements. The purpose of this paper is to compare the commercially available 'Calicheck' (Calcorp. Inc), used to assess linearity, versus the well-known source decay method, and also to show our results after performing several recommended quality control tests. The parameters that we wanted to evaluate were carried on using the Capintec CRC-15R and CRC-15 β radionuclide calibrators. The evaluated tests were: high voltage, display, zero adjust, background, reproducibility, source constancy, accuracy, precision and linearity. The first six tests were evaluated on the daily practice, here we analyzed the 2007 recorded data; and the last three were evaluated once a year. During the daily evaluation both calibrators performance were satisfactory comparing with the manufacture's requirements. The accuracy test show result within the ± 10% allowed for a field instrument. Precision performance is within the ± 1 % allowed. On the other hand, the linearity test shows that using the source decay method the relative coefficient is 0.9998, for both equipment and using the Calicheck the relative coefficient is 0.997. However, looking the percentage of error, during the 'Calicheck' test, its range goes from 0.0 % up to -25.35%, and using the source decay method, the range goes from 0.0 % up to -31.05 %, taking into account both instruments. Checking the 'Calicheck' results we can see that the results varying randomly, but using the source decay method the percentage of error increase as the source activity decrease. We conclude that both devices meet its manufactures requirements, in the case of the linearity using the decay method, decreasing the activity source, increasing the percentage of error, this may happen because of the equipment age. (author)

  1. Self-calibrating interferometer

    International Nuclear Information System (INIS)

    Nussmeier, T.A.

    1982-01-01

    A self-calibrating interferometer is disclosed which forms therein a pair of Michelson interferometers with one beam length of each Michelson interferometer being controlled by a common phase shifter. The transfer function measured from the phase shifter to either of a pair of detectors is sinusoidal with a full cycle for each half wavelength of phase shifter travel. The phase difference between these two sinusoidal detector outputs represents the optical phase difference between a path of known distance and a path of unknown distance

  2. Automation systems for radioimmunoassay

    International Nuclear Information System (INIS)

    Yamasaki, Paul

    1974-01-01

    The application of automation systems for radioimmunoassay (RIA) was discussed. Automated systems could be useful in the second step, of the four basic processes in the course of RIA, i.e., preparation of sample for reaction. There were two types of instrumentation, a semi-automatic pipete, and a fully automated pipete station, both providing for fast and accurate dispensing of the reagent or for the diluting of sample with reagent. Illustrations of the instruments were shown. (Mukohata, S.)

  3. Automatic and integrated micro-enzyme assay (AIμEA) platform for highly sensitive thrombin analysis via an engineered fluorescence protein-functionalized monolithic capillary column.

    Science.gov (United States)

    Lin, Lihua; Liu, Shengquan; Nie, Zhou; Chen, Yingzhuang; Lei, Chunyang; Wang, Zhen; Yin, Chao; Hu, Huiping; Huang, Yan; Yao, Shouzhuo

    2015-04-21

    Nowadays, large-scale screening for enzyme discovery, engineering, and drug discovery processes require simple, fast, and sensitive enzyme activity assay platforms with high integration and potential for high-throughput detection. Herein, a novel automatic and integrated micro-enzyme assay (AIμEA) platform was proposed based on a unique microreaction system fabricated by a engineered green fluorescence protein (GFP)-functionalized monolithic capillary column, with thrombin as an example. The recombinant GFP probe was rationally engineered to possess a His-tag and a substrate sequence of thrombin, which enable it to be immobilized on the monolith via metal affinity binding, and to be released after thrombin digestion. Combined with capillary electrophoresis-laser-induced fluorescence (CE-LIF), all the procedures, including thrombin injection, online enzymatic digestion in the microreaction system, and label-free detection of the released GFP, were integrated in a single electrophoretic process. By taking advantage of the ultrahigh loading capacity of the AIμEA platform and the CE automatic programming setup, one microreaction column was sufficient for many times digestion without replacement. The novel microreaction system showed significantly enhanced catalytic efficiency, about 30 fold higher than that of the equivalent bulk reaction. Accordingly, the AIμEA platform was highly sensitive with a limit of detection down to 1 pM of thrombin. Moreover, the AIμEA platform was robust and reliable to detect thrombin in human serum samples and its inhibition by hirudin. Hence, this AIμEA platform exhibits great potential for high-throughput analysis in future biological application, disease diagnostics, and drug screening.

  4. Essential role of cofilin-1 in regulating thrombin-induced RelA/p65 nuclear translocation and intercellular adhesion molecule 1 (ICAM-1) expression in endothelial cells.

    Science.gov (United States)

    Fazal, Fabeha; Bijli, Kaiser M; Minhajuddin, Mohd; Rein, Theo; Finkelstein, Jacob N; Rahman, Arshad

    2009-07-31

    Activation of RhoA/Rho-associated kinase (ROCK) pathway and the associated changes in actin cytoskeleton induced by thrombin are crucial for activation of NF-kappaB and expression of its target gene ICAM-1 in endothelial cells. However, the events acting downstream of RhoA/ROCK to mediate these responses remain unclear. Here, we show a central role of cofilin-1, an actin-binding protein that promotes actin depolymerization, in linking RhoA/ROCK pathway to dynamic alterations in actin cytoskeleton that are necessary for activation of NF-kappaB and thereby expression of ICAM-1 in these cells. Stimulation of human umbilical vein endothelial cells with thrombin resulted in Ser(3) phosphorylation/inactivation of cofilin and formation of actin stress fibers in a ROCK-dependent manner. RNA interference knockdown of cofilin-1 stabilized the actin filaments and inhibited thrombin- and RhoA-induced NF-kappaB activity. Similarly, constitutively inactive mutant of cofilin-1 (Cof1-S3D), known to stabilize the actin cytoskeleton, inhibited NF-kappaB activity by thrombin. Overexpression of wild type cofilin-1 or constitutively active cofilin-1 mutant (Cof1-S3A), known to destabilize the actin cytoskeleton, also impaired thrombin-induced NF-kappaB activity. Additionally, depletion of cofilin-1 was associated with a marked reduction in ICAM-1 expression induced by thrombin. The effect of cofilin-1 depletion on NF-kappaB activity and ICAM-1 expression occurred downstream of IkappaBalpha degradation and was a result of impaired RelA/p65 nuclear translocation and consequently, RelA/p65 binding to DNA. Together, these data show that cofilin-1 occupies a central position in RhoA-actin pathway mediating nuclear translocation of RelA/p65 and expression of ICAM-1 in endothelial cells.

  5. Essential Role of Cofilin-1 in Regulating Thrombin-induced RelA/p65 Nuclear Translocation and Intercellular Adhesion Molecule 1 (ICAM-1) Expression in Endothelial Cells*

    Science.gov (United States)

    Fazal, Fabeha; Bijli, Kaiser M.; Minhajuddin, Mohd; Rein, Theo; Finkelstein, Jacob N.; Rahman, Arshad

    2009-01-01

    Activation of RhoA/Rho-associated kinase (ROCK) pathway and the associated changes in actin cytoskeleton induced by thrombin are crucial for activation of NF-κB and expression of its target gene ICAM-1 in endothelial cells. However, the events acting downstream of RhoA/ROCK to mediate these responses remain unclear. Here, we show a central role of cofilin-1, an actin-binding protein that promotes actin depolymerization, in linking RhoA/ROCK pathway to dynamic alterations in actin cytoskeleton that are necessary for activation of NF-κB and thereby expression of ICAM-1 in these cells. Stimulation of human umbilical vein endothelial cells with thrombin resulted in Ser3 phosphorylation/inactivation of cofilin and formation of actin stress fibers in a ROCK-dependent manner. RNA interference knockdown of cofilin-1 stabilized the actin filaments and inhibited thrombin- and RhoA-induced NF-κB activity. Similarly, constitutively inactive mutant of cofilin-1 (Cof1-S3D), known to stabilize the actin cytoskeleton, inhibited NF-κB activity by thrombin. Overexpression of wild type cofilin-1 or constitutively active cofilin-1 mutant (Cof1-S3A), known to destabilize the actin cytoskeleton, also impaired thrombin-induced NF-κB activity. Additionally, depletion of cofilin-1 was associated with a marked reduction in ICAM-1 expression induced by thrombin. The effect of cofilin-1 depletion on NF-κB activity and ICAM-1 expression occurred downstream of IκBα degradation and was a result of impaired RelA/p65 nuclear translocation and consequently, RelA/p65 binding to DNA. Together, these data show that cofilin-1 occupies a central position in RhoA-actin pathway mediating nuclear translocation of RelA/p65 and expression of ICAM-1 in endothelial cells. PMID:19483084

  6. Dosimetry and Calibration Section

    International Nuclear Information System (INIS)

    Otto, T.

    1999-01-01

    The Dosimetry and Calibration Section fulfils two tasks within CERN's Radiation Protection Group: the Individual Dosimetry Service monitors more than 5000 persons potentially exposed to ionizing radiation on the CERN sites, and the Calibration Laboratory verifies throughout the year, at regular intervals, over 1000 instruments, monitors, and electronic dosimeters used by RP Group. The establishment of a Quality Assurance System for the Individual Dosimetry Service, a requirement of the new Swiss Ordinance for personal dosimetry, put a considerable workload on the section. Together with an external consultant it was decided to identify and then describe the different 'processes' of the routine work performed in the dosimetry service. The resulting Quality Manual was submitted to the Federal Office for Public Health in Bern in autumn. The CERN Individual Dosimetry Service will eventually be officially endorsed after a successful technical test in March 1999. On the technical side, the introduction of an automatic development machine for gamma films was very successful. It processes the dosimetric films without an operator being present, and its built-in regeneration mechanism keeps the concentration of the processing chemicals at a constant level

  7. Piezoelectric trace vapor calibrator

    International Nuclear Information System (INIS)

    Verkouteren, R. Michael; Gillen, Greg; Taylor, David W.

    2006-01-01

    The design and performance of a vapor generator for calibration and testing of trace chemical sensors are described. The device utilizes piezoelectric ink-jet nozzles to dispense and vaporize precisely known amounts of analyte solutions as monodisperse droplets onto a hot ceramic surface, where the generated vapors are mixed with air before exiting the device. Injected droplets are monitored by microscope with strobed illumination, and the reproducibility of droplet volumes is optimized by adjustment of piezoelectric wave form parameters. Complete vaporization of the droplets occurs only across a 10 deg. C window within the transition boiling regime of the solvent, and the minimum and maximum rates of trace analyte that may be injected and evaporated are determined by thermodynamic principles and empirical observations of droplet formation and stability. By varying solution concentrations, droplet injection rates, air flow, and the number of active nozzles, the system is designed to deliver--on demand--continuous vapor concentrations across more than six orders of magnitude (nominally 290 fg/l to 1.05 μg/l). Vapor pulses containing femtogram to microgram quantities of analyte may also be generated. Calibrated ranges of three explosive vapors at ng/l levels were generated by the device and directly measured by ion mobility spectrometry (IMS). These data demonstrate expected linear trends within the limited working range of the IMS detector and also exhibit subtle nonlinear behavior from the IMS measurement process

  8. A Simple Accelerometer Calibrator

    International Nuclear Information System (INIS)

    Salam, R A; Islamy, M R F; Khairurrijal; Munir, M M; Latief, H; Irsyam, M

    2016-01-01

    High possibility of earthquake could lead to the high number of victims caused by it. It also can cause other hazards such as tsunami, landslide, etc. In that case it requires a system that can examine the earthquake occurrence. Some possible system to detect earthquake is by creating a vibration sensor system using accelerometer. However, the output of the system is usually put in the form of acceleration data. Therefore, a calibrator system for accelerometer to sense the vibration is needed. In this study, a simple accelerometer calibrator has been developed using 12 V DC motor, optocoupler, Liquid Crystal Display (LCD) and AVR 328 microcontroller as controller system. The system uses the Pulse Wave Modulation (PWM) form microcontroller to control the motor rotational speed as response to vibration frequency. The frequency of vibration was read by optocoupler and then those data was used as feedback to the system. The results show that the systems could control the rotational speed and the vibration frequencies in accordance with the defined PWM. (paper)

  9. Laboratory Automation and Middleware.

    Science.gov (United States)

    Riben, Michael

    2015-06-01

    The practice of surgical pathology is under constant pressure to deliver the highest quality of service, reduce errors, increase throughput, and decrease turnaround time while at the same time dealing with an aging workforce, increasing financial constraints, and economic uncertainty. Although not able to implement total laboratory automation, great progress continues to be made in workstation automation in all areas of the pathology laboratory. This report highlights the benefits and challenges of pathology automation, reviews middleware and its use to facilitate automation, and reviews the progress so far in the anatomic pathology laboratory. Copyright © 2015 Elsevier Inc. All rights reserved.

  10. Automated cloning methods.; TOPICAL

    International Nuclear Information System (INIS)

    Collart, F.

    2001-01-01

    Argonne has developed a series of automated protocols to generate bacterial expression clones by using a robotic system designed to be used in procedures associated with molecular biology. The system provides plate storage, temperature control from 4 to 37 C at various locations, and Biomek and Multimek pipetting stations. The automated system consists of a robot that transports sources from the active station on the automation system. Protocols for the automated generation of bacterial expression clones can be grouped into three categories (Figure 1). Fragment generation protocols are initiated on day one of the expression cloning procedure and encompass those protocols involved in generating purified coding region (PCR)

  11. Complacency and Automation Bias in the Use of Imperfect Automation.

    Science.gov (United States)

    Wickens, Christopher D; Clegg, Benjamin A; Vieane, Alex Z; Sebok, Angelia L

    2015-08-01

    We examine the effects of two different kinds of decision-aiding automation errors on human-automation interaction (HAI), occurring at the first failure following repeated exposure to correctly functioning automation. The two errors are incorrect advice, triggering the automation bias, and missing advice, reflecting complacency. Contrasts between analogous automation errors in alerting systems, rather than decision aiding, have revealed that alerting false alarms are more problematic to HAI than alerting misses are. Prior research in decision aiding, although contrasting the two aiding errors (incorrect vs. missing), has confounded error expectancy. Participants performed an environmental process control simulation with and without decision aiding. For those with the aid, automation dependence was created through several trials of perfect aiding performance, and an unexpected automation error was then imposed in which automation was either gone (one group) or wrong (a second group). A control group received no automation support. The correct aid supported faster and more accurate diagnosis and lower workload. The aid failure degraded all three variables, but "automation wrong" had a much greater effect on accuracy, reflecting the automation bias, than did "automation gone," reflecting the impact of complacency. Some complacency was manifested for automation gone, by a longer latency and more modest reduction in accuracy. Automation wrong, creating the automation bias, appears to be a more problematic form of automation error than automation gone, reflecting complacency. Decision-aiding automation should indicate its lower degree of confidence in uncertain environments to avoid the automation bias. © 2015, Human Factors and Ergonomics Society.

  12. Monitoring low molecular weight heparins at therapeutic levels: dose-responses of, and correlations and differences between aPTT, anti-factor Xa and thrombin generation assays.

    Directory of Open Access Journals (Sweden)

    Owain Thomas

    Full Text Available Low molecular weight heparins (LMWH's are used to prevent and treat thrombosis. Tests for monitoring LMWH's include anti-factor Xa (anti-FXa, activated partial thromboplastin time (aPTT and thrombin generation. Anti-FXa is the current gold standard despite LMWH's varying affinities for FXa and thrombin.To examine the effects of two different LMWH's on the results of 4 different aPTT-tests, anti-FXa activity and thrombin generation and to assess the tests' concordance.Enoxaparin and tinzaparin were added ex-vivo in concentrations of 0.0, 0.5, 1.0 and 1.5 anti-FXa international units (IU/mL, to blood from 10 volunteers. aPTT was measured using two whole blood methods (Free oscillation rheometry (FOR and Hemochron Jr (HCJ and an optical plasma method using two different reagents (ActinFSL and PTT-Automat. Anti-FXa activity was quantified using a chromogenic assay. Thrombin generation (Endogenous Thrombin Potential, ETP was measured on a Ceveron Alpha instrument using the TGA RB and more tissue-factor rich TGA RC reagents.Methods' mean aPTT at 1.0 IU/mL LMWH varied between 54s (SD 11 and 69s (SD 14 for enoxaparin and between 101s (SD 21 and 140s (SD 28 for tinzaparin. ActinFSL gave significantly shorter aPTT results. aPTT and anti-FXa generally correlated well. ETP as measured with the TGA RC reagent but not the TGA RB reagent showed an inverse exponential relationship to the concentration of LMWH. The HCJ-aPTT results had the weakest correlation to anti-FXa and thrombin generation (Rs0.62-0.87, whereas the other aPTT methods had similar correlation coefficients (Rs0.80-0.92.aPTT displays a linear dose-response to LMWH. There is variation between aPTT assays. Tinzaparin increases aPTT and decreases thrombin generation more than enoxaparin at any given level of anti-FXa activity, casting doubt on anti-FXa's present gold standard status. Thrombin generation with tissue factor-rich activator is a promising method for monitoring LMWH's.

  13. Will Automated Vehicles Negatively Impact Traffic Flow?

    Directory of Open Access Journals (Sweden)

    S. C. Calvert

    2017-01-01

    Full Text Available With low-level vehicle automation already available, there is a necessity to estimate its effects on traffic flow, especially if these could be negative. A long gradual transition will occur from manual driving to automated driving, in which many yet unknown traffic flow dynamics will be present. These effects have the potential to increasingly aid or cripple current road networks. In this contribution, we investigate these effects using an empirically calibrated and validated simulation experiment, backed up with findings from literature. We found that low-level automated vehicles in mixed traffic will initially have a small negative effect on traffic flow and road capacities. The experiment further showed that any improvement in traffic flow will only be seen at penetration rates above 70%. Also, the capacity drop appeared to be slightly higher with the presence of low-level automated vehicles. The experiment further investigated the effect of bottleneck severity and truck shares on traffic flow. Improvements to current traffic models are recommended and should include a greater detail and understanding of driver-vehicle interaction, both in conventional and in mixed traffic flow. Further research into behavioural shifts in driving is also recommended due to limited data and knowledge of these dynamics.

  14. Genetic algorithm for building envelope calibration

    International Nuclear Information System (INIS)

    Ramos Ruiz, Germán; Fernández Bandera, Carlos; Gómez-Acebo Temes, Tomás; Sánchez-Ostiz Gutierrez, Ana

    2016-01-01

    Highlights: • Calibration methodology using Multi-Objective Genetic Algorithm (NSGA-II). • Uncertainty analysis formulas implemented directly in EnergyPlus. • The methodology captures the heat dynamic of the building with a high level of accuracy. • Reduction in the number of parameters involved due to sensitivity analysis. • Cost-effective methodology using temperature sensors only. - Abstract: Buildings today represent 40% of world primary energy consumption and 24% of greenhouse gas emissions. In our society there is growing interest in knowing precisely when and how energy consumption occurs. This means that consumption measurement and verification plans are well-advanced. International agencies such as Efficiency Valuation Organization (EVO) and International Performance Measurement and Verification Protocol (IPMVP) have developed methodologies to quantify savings. This paper presents a methodology to accurately perform automated envelope calibration under option D (calibrated simulation) of IPMVP – vol. 1. This is frequently ignored because of its complexity, despite being more flexible and accurate in assessing the energy performance of a building. A detailed baseline energy model is used, and by means of a metaheuristic technique achieves a highly reliable and accurate Building Energy Simulation (BES) model suitable for detailed analysis of saving strategies. In order to find this BES model a Genetic Algorithm (NSGA-II) is used, together with a highly efficient engine to stimulate the objective, thus permitting rapid achievement of the goal. The result is a BES model that broadly captures the heat dynamic behaviour of the building. The model amply fulfils the parameters demanded by ASHRAE and EVO under option D.

  15. Host defense peptides of thrombin modulate inflammation and coagulation in endotoxin-mediated shock and Pseudomonas aeruginosa sepsis.

    Science.gov (United States)

    Kalle, Martina; Papareddy, Praveen; Kasetty, Gopinath; Mörgelin, Matthias; van der Plas, Mariena J A; Rydengård, Victoria; Malmsten, Martin; Albiger, Barbara; Schmidtchen, Artur

    2012-01-01

    Gram-negative sepsis is accompanied by a disproportionate innate immune response and excessive coagulation mainly induced by endotoxins released from bacteria. Due to rising antibiotic resistance and current lack of other effective treatments there is an urgent need for new therapies. We here present a new treatment concept for sepsis and endotoxin-mediated shock, based on host defense peptides from the C-terminal part of human thrombin, found to have a broad and inhibitory effect on multiple sepsis pathologies. Thus, the peptides abrogate pro-inflammatory cytokine responses to endotoxin in vitro and in vivo. Furthermore, they interfere with coagulation by modulating contact activation and tissue factor-mediated clotting in vitro, leading to normalization of coagulation responses in vivo, a previously unknown function of host defense peptides. In a mouse model of Pseudomonas aeruginosa sepsis, the peptide GKY25, while mediating a modest antimicrobial effect, significantly inhibited the pro-inflammatory response, decreased fibrin deposition and leakage in the lungs, as well as reduced mortality. Taken together, the capacity of such thrombin-derived peptides to simultaneously modulate bacterial levels, pro-inflammatory responses, and coagulation, renders them attractive therapeutic candidates for the treatment of invasive infections and sepsis.

  16. Ultrasound-Guided Thrombin Injection Is a Safe and Effective Treatment for Femoral Artery Pseudoaneurysm in the Morbidly Obese.

    Science.gov (United States)

    Yoo, Taehwan; Starr, Jean E; Go, Michael R; Vaccaro, Patrick S; Satiani, Bhagwan; Haurani, Mounir J

    2017-08-01

    Ultrasound-guided thrombin injection (UGTI) is a well-established practice for the treatment of femoral artery pseudoaneurysm. This procedure is highly successful but dependent on appropriate pseudoaneurysm anatomy and adequate ultrasound visualization. Morbid obesity can present a significant technical challenge due to increased groin adiposity, resulting in poor visualization of critical structures needed to safely perform the procedure. We aim to evaluate the safety and efficacy of UGTI to treat femoral artery pseudoaneurysm in the morbidly obese. This is a retrospective cohort study in which all patients who underwent UGTI at The Ohio State University Ross Heart Hospital from 2009 to 2014 were analyzed for patient characteristics and stratified by body mass index (BMI). Patients with BMI ≥ 35 were considered morbidly obese and were compared to patients with a BMI injection. There were 41 nonmorbidly obese and 13 morbidly obese patients. Mean age was 64.5 years. The cohort was 44.4% male. There were 6 failures, of which 1 underwent successful repeat injection and 5 underwent open surgical repair. There was no statistically significant difference in failure between nonmorbidly obese and morbidly obese patients (9.8% vs 15.4%, P = .45). There were no embolic/thrombotic complications. Ultrasound-guided thrombin injection is a safe and effective therapy in the morbidly obese for the treatment of femoral artery pseudoaneurysm. In the hands of experienced sonographers and surgeons with adequate visualization of the pseudoaneurysm sac, UGTI should remain a standard therapy in the morbidly obese.

  17. Inhibition of Cellular Adhesion by Immunological Targeting of Osteopontin Neoepitopes Generated through Matrix Metalloproteinase and Thrombin Cleavage.

    Science.gov (United States)

    Jürets, Alexander; Le Bras, Marie; Staffler, Günther; Stein, Gesine; Leitner, Lukas; Neuhofer, Angelika; Tardelli, Matteo; Turkof, Edvin; Zeyda, Maximilian; Stulnig, Thomas M

    2016-01-01

    Osteopontin (OPN), a secreted protein involved in inflammatory processes and cancer, induces cell adhesion, migration, and activation of inflammatory pathways in various cell types. Cells bind OPN via integrins at a canonical RGD region in the full length form as well as to a contiguous cryptic site that some have shown is unmasked upon thrombin or matrix metalloproteinase cleavage. Thus, the adhesive capacity of osteopontin is enhanced by proteolytic cleavage that may occur in inflammatory conditions such as obesity, atherosclerosis, rheumatoid arthritis, tumor growth and metastasis. Our aim was to inhibit cellular adhesion to recombinant truncated proteins that correspond to the N-terminal cleavage products of thrombin- or matrix metalloproteinase-cleaved OPN in vitro. We specifically targeted the cryptic integrin binding site with monoclonal antibodies and antisera induced by peptide immunization of mice. HEK 293 cells adhered markedly stronger to truncated OPN proteins than to full length OPN. Without affecting cell binding to the full length form, the raised monoclonal antibodies specifically impeded cellular adhesion to the OPN fragments. Moreover, we show that the peptides used for immunization were able to induce antisera, which impeded adhesion either to all OPN forms, including the full-length form, or selectively to the corresponding truncated recombinant proteins. In conclusion, we developed immunological tools to selectively target functional properties of protease-cleaved OPN forms, which could find applications in treatment and prevention of various inflammatory diseases and cancers.

  18. Host defense peptides of thrombin modulate inflammation and coagulation in endotoxin-mediated shock and Pseudomonas aeruginosa sepsis.

    Directory of Open Access Journals (Sweden)

    Martina Kalle

    Full Text Available Gram-negative sepsis is accompanied by a disproportionate innate immune response and excessive coagulation mainly induced by endotoxins released from bacteria. Due to rising antibiotic resistance and current lack of other effective treatments there is an urgent need for new therapies. We here present a new treatment concept for sepsis and endotoxin-mediated shock, based on host defense peptides from the C-terminal part of human thrombin, found to have a broad and inhibitory effect on multiple sepsis pathologies. Thus, the peptides abrogate pro-inflammatory cytokine responses to endotoxin in vitro and in vivo. Furthermore, they interfere with coagulation by modulating contact activation and tissue factor-mediated clotting in vitro, leading to normalization of coagulation responses in vivo, a previously unknown function of host defense peptides. In a mouse model of Pseudomonas aeruginosa sepsis, the peptide GKY25, while mediating a modest antimicrobial effect, significantly inhibited the pro-inflammatory response, decreased fibrin deposition and leakage in the lungs, as well as reduced mortality. Taken together, the capacity of such thrombin-derived peptides to simultaneously modulate bacterial levels, pro-inflammatory responses, and coagulation, renders them attractive therapeutic candidates for the treatment of invasive infections and sepsis.

  19. Isolation and characterization of a serine proteinase with thrombin-like activity from the venom of the snake Bothrops asper

    Directory of Open Access Journals (Sweden)

    A.V Pérez

    2008-01-01

    Full Text Available A serine proteinase with thrombin-like activity was isolated from the venom of the Central American pit viper Bothrops asper. Isolation was performed by a combination of affinity chromatography on aminobenzamidine-Sepharose and ion-exchange chromatography on DEAE-Sepharose. The enzyme accounts for approximately 0.13% of the venom dry weight and has a molecular mass of 32 kDa as determined by SDS-PAGE, and of 27 kDa as determined by MALDI-TOF mass spectrometry. Its partial amino acid sequence shows high identity with snake venom serine proteinases and a complete identity with a cDNA clone previously sequenced from this species. The N-terminal sequence of the enzyme is VIGGDECNINEHRSLVVLFXSSGFL CAGTLVQDEWVLTAANCDSKNFQ. The enzyme induces clotting of plasma (minimum coagulant dose = 4.1 µg and fibrinogen (minimum coagulant dose = 4.2 µg in vitro, and promotes defibrin(ogenation in vivo (minimum defibrin(ogenating dose = 1.0 µg. In addition, when injected intravenously in mice at doses of 5 and 10 µg, it induces a series of behavioral changes, i.e., loss of the righting reflex, opisthotonus, and intermittent rotations over the long axis of the body, which closely resemble the `gyroxin-like' effect induced by other thrombin-like enzymes from snake venoms.

  20. On chromatic and geometrical calibration

    DEFF Research Database (Denmark)

    Folm-Hansen, Jørgen

    1999-01-01

    The main subject of the present thesis is different methods for the geometrical and chromatic calibration of cameras in various environments. For the monochromatic issues of the calibration we present the acquisition of monochrome images, the classic monochrome aberrations and the various sources...... the correct interpolation method is described. For the chromatic issues of calibration we present the acquisition of colour and multi-spectral images, the chromatic aberrations and the various lens/camera based non-uniformities of the illumination of the image plane. It is described how the monochromatic...... to design calibration targets for both geometrical and chromatic calibration are described. We present some possible systematical errors on the detection of the objects in the calibration targets, if viewed in a non orthogonal angle, if the intensities are uneven or if the image blurring is uneven. Finally...

  1. Development of portable flow calibrator

    International Nuclear Information System (INIS)

    Akiyama, Kiyomitsu; Iijima, Nobuo

    1995-01-01

    In the nuclear facilities, air sniffer system is often utilized to evaluate atmospheric concentration of radioactivity in the working environment. The system collects airborne dust on the filter during some sampling period. In this method, total air flow during the sampling period is an important parameter to evaluate the radioactivity concentration correctly. Therefore, calibration for the flow meter of air sniffer system must be done periodically according to Japan Industry Standards (JIS). As we have had to available device to calibrate the flow meter in the working area, we had to remove the flow meters from the installed place and carry them to another place where calibration can be made. This work required a great deal of labor. Now we have developed a portable flow calibrator for air sniffer system which enables us to make in-site calibration of the flow meter in the working area more easily. This report describes the outline of portable flow calibrator and it's experimental results. (author)

  2. Quality Management and Calibration

    Science.gov (United States)

    Merkus, Henk G.

    Good specification of a product’s performance requires adequate characterization of relevant properties. Particulate products are usually characterized by some PSD, shape or porosity parameter(s). For proper characterization, adequate sampling, dispersion, and measurement procedures should be available or developed and skilful personnel should use appropriate, well-calibrated/qualified equipment. The characterization should be executed, in agreement with customers, in a wellorganized laboratory. All related aspects should be laid down in a quality handbook. The laboratory should provide proof for its capability to perform the characterization of stated products and/or reference materials within stated confidence limits. This can be done either by internal validation and audits or by external GLP accreditation.

  3. SURF Model Calibration Strategy

    Energy Technology Data Exchange (ETDEWEB)

    Menikoff, Ralph [Los Alamos National Lab. (LANL), Los Alamos, NM (United States)

    2017-03-10

    SURF and SURFplus are high explosive reactive burn models for shock initiation and propagation of detonation waves. They are engineering models motivated by the ignition & growth concept of high spots and for SURFplus a second slow reaction for the energy release from carbon clustering. A key feature of the SURF model is that there is a partial decoupling between model parameters and detonation properties. This enables reduced sets of independent parameters to be calibrated sequentially for the initiation and propagation regimes. Here we focus on a methodology for tting the initiation parameters to Pop plot data based on 1-D simulations to compute a numerical Pop plot. In addition, the strategy for tting the remaining parameters for the propagation regime and failure diameter is discussed.

  4. Calibration of Nanopositioning Stages

    Directory of Open Access Journals (Sweden)

    Ning Tan

    2015-12-01

    Full Text Available Accuracy is one of the most important criteria for the performance evaluation of micro- and nanorobots or systems. Nanopositioning stages are used to achieve the high positioning resolution and accuracy for a wide and growing scope of applications. However, their positioning accuracy and repeatability are not well known and difficult to guarantee, which induces many drawbacks for many applications. For example, in the mechanical characterisation of biological samples, it is difficult to perform several cycles in a repeatable way so as not to induce negative influences on the study. It also prevents one from controlling accurately a tool with respect to a sample without adding additional sensors for closed loop control. This paper aims at quantifying the positioning repeatability and accuracy based on the ISO 9283:1998 standard, and analyzing factors influencing positioning accuracy onto a case study of 1-DoF (Degree-of-Freedom nanopositioning stage. The influence of thermal drift is notably quantified. Performances improvement of the nanopositioning stage are then investigated through robot calibration (i.e., open-loop approach. Two models (static and adaptive models are proposed to compensate for both geometric errors and thermal drift. Validation experiments are conducted over a long period (several days showing that the accuracy of the stage is improved from typical micrometer range to 400 nm using the static model and even down to 100 nm using the adaptive model. In addition, we extend the 1-DoF calibration to multi-DoF with a case study of a 2-DoF nanopositioning robot. Results demonstrate that the model efficiently improved the 2D accuracy from 1400 nm to 200 nm.

  5. Radiological Calibration and Standards Facility

    Data.gov (United States)

    Federal Laboratory Consortium — PNNL maintains a state-of-the-art Radiological Calibration and Standards Laboratory on the Hanford Site at Richland, Washington. Laboratory staff provide expertise...

  6. Portable compact multifunction IR calibrator

    International Nuclear Information System (INIS)

    Wyatt, C.L.; Jacobsen, L.; Steed, A.

    1988-01-01

    A compact portable multifunction calibrator designed for future sensor systems is described which enables a linearity calibration for all detectors simultaneously using a near small-area source, a high-resolution mapping of the focal plane with 10 microrad setability and with a blur of less than 100 microrad, system spectral response calibration (radiometer) using a Michelson interferometer source, relative spectral response (spectrometer) using high-temperature external commercial blackbody simulators, and an absolute calibration using an internal low-temperature extended-area source. 5 references

  7. Automated System Marketplace 1994.

    Science.gov (United States)

    Griffiths, Jose-Marie; Kertis, Kimberly

    1994-01-01

    Reports results of the 1994 Automated System Marketplace survey based on responses from 60 vendors. Highlights include changes in the library automation marketplace; estimated library systems revenues; minicomputer and microcomputer-based systems; marketplace trends; global markets and mergers; research needs; new purchase processes; and profiles…

  8. Automation in Warehouse Development

    NARCIS (Netherlands)

    Hamberg, R.; Verriet, J.

    2012-01-01

    The warehouses of the future will come in a variety of forms, but with a few common ingredients. Firstly, human operational handling of items in warehouses is increasingly being replaced by automated item handling. Extended warehouse automation counteracts the scarcity of human operators and

  9. Order Division Automated System.

    Science.gov (United States)

    Kniemeyer, Justin M.; And Others

    This publication was prepared by the Order Division Automation Project staff to fulfill the Library of Congress' requirement to document all automation efforts. The report was originally intended for internal use only and not for distribution outside the Library. It is now felt that the library community at-large may have an interest in the…

  10. Automate functional testing

    Directory of Open Access Journals (Sweden)

    Ramesh Kalindri

    2014-06-01

    Full Text Available Currently, software engineers are increasingly turning to the option of automating functional tests, but not always have successful in this endeavor. Reasons range from low planning until over cost in the process. Some principles that can guide teams in automating these tests are described in this article.

  11. Automation and robotics

    Science.gov (United States)

    Montemerlo, Melvin

    1988-01-01

    The Autonomous Systems focus on the automation of control systems for the Space Station and mission operations. Telerobotics focuses on automation for in-space servicing, assembly, and repair. The Autonomous Systems and Telerobotics each have a planned sequence of integrated demonstrations showing the evolutionary advance of the state-of-the-art. Progress is briefly described for each area of concern.

  12. Automating the Small Library.

    Science.gov (United States)

    Skapura, Robert

    1987-01-01

    Discusses the use of microcomputers for automating school libraries, both for entire systems and for specific library tasks. Highlights include available library management software, newsletters that evaluate software, constructing an evaluation matrix, steps to consider in library automation, and a brief discussion of computerized card catalogs.…

  13. BXS Re-calibration

    International Nuclear Information System (INIS)

    Welch, J.

    2010-01-01

    Early in the commissioning it was noticed by Cecile Limborg that the calibration of the BXS spectrometer magnet seemed to be different from the strength of the BX01/BX02 magnets. First the BX01/BX02 currents were adjusted to 135 MeV and the beam energy was adjusted to make the horizontal orbit flat. Then BX01/BX02 magnets were switched off and BXS was adjusted to make the horizontal orbit in the spectrometer line flat, without changing the energy of the beam. The result was that about 140-141 MeV were required on the BXS magnet. This measurement was repeated several times by others with the same results. It was not clear what was causing the error: magnet strength or layout. A position error of about 19 mm of the BXS magnet could explain the difference. Because there was a significant misalignment of the vacuum chamber in the BXS line, the alignment of the whole spectrometer line was checked. The vacuum chamber was corrected, but the magnets were found to be in the proper alignment. So we were left with one (or conceivably two) magnet calibration errors. Because BXS is a wedged shaped magnet, the bend angle depends on the horizontal position of the incoming beam. As mentioned, an offset of the beam position of 19 mm would increase or decrease the bend angle roughly by the ratio of 135/141. The figure of 19 mm is special and caused a considerable confusion during the design and measurement of the BXS magnet. This is best illustrated in Figure 1 which was taken out of the BXS Traveler document. The distance between the horizontal midplanes of the poles and the apex of the beam path was chosen to be 19 mm so the beam is close to the good field region throughout its entire path. Thus it seemed possible that there was an error that resulted in the beam not being on this trajectory, or conversely, that the magnetic measurements were done on the wrong trajectory and the magnet was then mis-calibrated. Mechanical measurements of the vacuum chamber made in the tunnel

  14. Preparatory research to develop an operational method to calibrate airborne sensor data using a network of ground calibration sites

    International Nuclear Information System (INIS)

    Milton, E.J.; Smith, G.M.; Lawless, K.P.

    1996-01-01

    The objective of the research is to develop an operational method to convert airborne spectral radiance data to reflectance using a number of well-characterized ground calibration sites located around the UK. The study is in three phases. First, a pilot study has been conducted at a disused airfield in southern England to test the feasibility of the open-quote empirical line close-quote method of sensor calibration. The second phase is developing methods to predict temporal changes in the bidirectional reflectance of ground calibration sites. The final phase of the project will look at methods to extend such calibrations spatially. This paper presents some results from the first phase of this study. The viability of the empirical line method of correction is shown to depend upon the use of ground targets whose in-band reflectance encompasses that of the targets of interest in the spectral band(s) concerned. The experimental design for the second phase of the study, in which methods to predict temporal trends in the bidirectional reflectance of these sites will be developed, is discussed. Finally, it is planned to develop an automated method of searching through Landsat TM data for the UK to identify a number of candidate ground calibration sites for which the model can be tested. 11 refs., 5 figs., 5 tabs

  15. Automated model building

    CERN Document Server

    Caferra, Ricardo; Peltier, Nicholas

    2004-01-01

    This is the first book on automated model building, a discipline of automated deduction that is of growing importance Although models and their construction are important per se, automated model building has appeared as a natural enrichment of automated deduction, especially in the attempt to capture the human way of reasoning The book provides an historical overview of the field of automated deduction, and presents the foundations of different existing approaches to model construction, in particular those developed by the authors Finite and infinite model building techniques are presented The main emphasis is on calculi-based methods, and relevant practical results are provided The book is of interest to researchers and graduate students in computer science, computational logic and artificial intelligence It can also be used as a textbook in advanced undergraduate courses

  16. Automation in Immunohematology

    Directory of Open Access Journals (Sweden)

    Meenu Bajpai

    2012-01-01

    Full Text Available There have been rapid technological advances in blood banking in South Asian region over the past decade with an increasing emphasis on quality and safety of blood products. The conventional test tube technique has given way to newer techniques such as column agglutination technique, solid phase red cell adherence assay, and erythrocyte-magnetized technique. These new technologies are adaptable to automation and major manufacturers in this field have come up with semi and fully automated equipments for immunohematology tests in the blood bank. Automation improves the objectivity and reproducibility of tests. It reduces human errors in patient identification and transcription errors. Documentation and traceability of tests, reagents and processes and archiving of results is another major advantage of automation. Shifting from manual methods to automation is a major undertaking for any transfusion service to provide quality patient care with lesser turnaround time for their ever increasing workload. This article discusses the various issues involved in the process.

  17. Automation in Warehouse Development

    CERN Document Server

    Verriet, Jacques

    2012-01-01

    The warehouses of the future will come in a variety of forms, but with a few common ingredients. Firstly, human operational handling of items in warehouses is increasingly being replaced by automated item handling. Extended warehouse automation counteracts the scarcity of human operators and supports the quality of picking processes. Secondly, the development of models to simulate and analyse warehouse designs and their components facilitates the challenging task of developing warehouses that take into account each customer’s individual requirements and logistic processes. Automation in Warehouse Development addresses both types of automation from the innovative perspective of applied science. In particular, it describes the outcomes of the Falcon project, a joint endeavour by a consortium of industrial and academic partners. The results include a model-based approach to automate warehouse control design, analysis models for warehouse design, concepts for robotic item handling and computer vision, and auton...

  18. A global model for residential energy use: Uncertainty in calibration to regional data

    International Nuclear Information System (INIS)

    van Ruijven, Bas; van Vuuren, Detlef P.; de Vries, Bert; van der Sluijs, Jeroen P.

    2010-01-01

    Uncertainties in energy demand modelling allow for the development of different models, but also leave room for different calibrations of a single model. We apply an automated model calibration procedure to analyse calibration uncertainty of residential sector energy use modelling in the TIMER 2.0 global energy model. This model simulates energy use on the basis of changes in useful energy intensity, technology development (AEEI) and price responses (PIEEI). We find that different implementations of these factors yield behavioural model results. Model calibration uncertainty is identified as influential source for variation in future projections: amounting 30% to 100% around the best estimate. Energy modellers should systematically account for this and communicate calibration uncertainty ranges. (author)

  19. Cryogenic thermometer calibration system using a helium cooling loop and a temperature controller [for LHC magnets

    CERN Document Server

    Chanzy, E; Thermeau, J P; Bühler, S; Joly, C; Casas-Cubillos, J; Balle, C

    1998-01-01

    The IPN-Orsay and CERN are designing in close collaboration a fully automated cryogenic thermometer calibration facility which will calibrate in 3 years 10,000 cryogenic thermometers required for the Large Hadron Collider (LHC) operation. A reduced-scale model of the calibration facility has been developed, which enables the calibration of ten thermometers by comparison with two rhodium-iron standard thermometers in the 1.8 K to 300 K temperature range under vacuum conditions. The particular design, based on a helium cooling loop and an electrical temperature controller, gives good dynamic performances. This paper describes the experimental set-up and the data acquisition system. Results of experimental runs are also presented along with the estimated global accuracy for the calibration. (3 refs).

  20. Genetic Algorithm Calibration of Probabilistic Cellular Automata for Modeling Mining Permit Activity

    Science.gov (United States)

    Louis, S.J.; Raines, G.L.

    2003-01-01

    We use a genetic algorithm to calibrate a spatially and temporally resolved cellular automata to model mining activity on public land in Idaho and western Montana. The genetic algorithm searches through a space of transition rule parameters of a two dimensional cellular automata model to find rule parameters that fit observed mining activity data. Previous work by one of the authors in calibrating the cellular automaton took weeks - the genetic algorithm takes a day and produces rules leading to about the same (or better) fit to observed data. These preliminary results indicate that genetic algorithms are a viable tool in calibrating cellular automata for this application. Experience gained during the calibration of this cellular automata suggests that mineral resource information is a critical factor in the quality of the results. With automated calibration, further refinements of how the mineral-resource information is provided to the cellular automaton will probably improve our model.

  1. Exposure-rate calibration using large-area calibration pads

    International Nuclear Information System (INIS)

    Novak, E.F.

    1988-09-01

    The US Department of Energy (DOE) Office of Remedial Action and Waste Technology established the Technical Measurements Center (TMC) at the DOE Grand Junction Projects Office (GJPO) in Grand Junction, Colorado, to standardize, calibrate, and compare measurements made in support of DOE remedial action programs. A set of large-area, radioelement-enriched concrete pads was constructed by the DOE in 1978 at the Walker Field Airport in Grand Junction for use as calibration standards for airborne gamma-ray spectrometer systems. The use of these pads was investigated by the TMC as potential calibration standards for portable scintillometers employed in measuring gamma-ray exposure rates at Uranium Mill Tailings Remedial Action (UMTRA) project sites. Data acquired on the pads using a pressurized ionization chamber (PIC) and three scintillometers are presented as an illustration of an instrumental calibration. Conclusions and recommended calibration procedures are discussed, based on the results of these data

  2. Automated complex spectra processing of actinide α-radiation

    International Nuclear Information System (INIS)

    Anichenkov, S.V.; Popov, Yu.S.; Tselishchev, I.V.; Mishenev, V.B.; Timofeev, G.A.

    1989-01-01

    Earlier described algorithms of automated processing of complex α - spectra of actinides with the use of Ehlektronika D3-28 computer line, connected with ICA-070 multichannel amplitude pulse analyzer, were realized. The developed program enables to calculated peak intensity and the relative isotope content, to conduct energy calibration of spectra, to calculate peak center of gravity and energy resolution, to perform integral counting in particular part of the spectrum. Error of the method of automated processing depens on the degree of spectrum complication and lies within the limits of 1-12%. 8 refs.; 4 figs.; 2 tabs

  3. Automated phased array ultrasonic inspection system for rail wheel sets

    International Nuclear Information System (INIS)

    Grosser, Paul; Weiland, M.G.

    2013-01-01

    This paper covers the design, system automation, calibration and validation of an automated ultrasonic system for the inspection of new and in service wheel set assemblies from diesel-electric locomotives and gondola cars. This system uses Phased Array (PA) transducers for flaw detection and Electro-Magnetic Acoustic Transducers (EMAT) for the measurement of residual stress. The system collects, analyses, evaluates and categorizes the wheel sets automatically. This data is archived for future comparison and trending. It is also available for export to a portal lathe for increased efficiency and accuracy of machining, therefore allowing prolonged wheel life.

  4. cGMP and nitric oxide modulate thrombin-induced endothelial permeability : Regulation via different pathways in human aortic and umbilical vein endothelial cells

    NARCIS (Netherlands)

    Draijer, R.; Atsma, D.E.; Laarse, A. van der; Hinsbergh, V.W.M. van

    1995-01-01

    Previous studies have demonstrated that cGMP and cAMP reduce the endothelial permeability for fluids and macromolecules when the endothelial permeability is increased by thrombin. In this study, we have investigated the mechanism by which cGMP improves the endothelial barrier function and examined

  5. Critical role of non-muscle myosin light chain kinase in thrombin-induced endothelial cell inflammation and lung PMN infiltration.

    Science.gov (United States)

    Fazal, Fabeha; Bijli, Kaiser M; Murrill, Matthew; Leonard, Antony; Minhajuddin, Mohammad; Anwar, Khandaker N; Finkelstein, Jacob N; Watterson, D Martin; Rahman, Arshad

    2013-01-01

    The pathogenesis of acute lung injury (ALI) involves bidirectional cooperation and close interaction between inflammatory and coagulation pathways. A key molecule linking coagulation and inflammation is the procoagulant thrombin, a serine protease whose concentration is elevated in plasma and lavage fluids of patients with ALI and acute respiratory distress syndrome (ARDS). However, little is known about the mechanism by which thrombin contributes to lung inflammatory response. In this study, we developed a new mouse model that permits investigation of lung inflammation associated with intravascular coagulation. Using this mouse model and in vitro approaches, we addressed the role of non-muscle myosin light chain kinase (nmMLCK) in thrombin-induced endothelial cell (EC) inflammation and lung neutrophil (PMN) infiltration. Our in vitro experiments revealed a key role of nmMLCK in ICAM-1 expression by its ability to control nuclear translocation and transcriptional capacity of RelA/p65 in EC. When subjected to intraperitoneal thrombin challenge, wild type mice showed a marked increase in lung PMN infiltration via expression of ICAM-1. However, these responses were markedly attenuated in mice deficient in nmMLCK. These results provide mechanistic insight into lung inflammatory response associated with intravascular coagulation and identify nmMLCK as a critical target for modulation of lung inflammation.

  6. Critical role of FcR gamma-chain, LAT, PLCgamma2 and thrombin in arteriolar thrombus formation upon mild, laser-induced endothelial injury in vivo.

    Science.gov (United States)

    Kalia, Neena; Auger, Jocelyn M; Atkinson, Ben; Watson, Steve P

    2008-05-01

    The role of collagen receptor complex GPVI-FcR gamma-chain, PLCgamma2 and LAT in laser-induced thrombosis is unclear. Controversy surrounds whether collagen is exposed in this model or whether thrombosis is dependent on thrombin. This study hypothesized that collagen exposure plays a critical role in thrombus formation in this model, which was tested by investigating contributions of FcR gamma-chain, LAT, PLCgamma2 and thrombin. Thrombi were monitored using intravital microscopy in anesthetized wild-type and FcR gamma-chain, LAT and PLCgamma2 knockout mice. Hirudin (thrombin inhibitor) was administered to wild-type and FcR gamma-chain knockout mice. Significantly reduced thrombus formation was observed in FcR gamma-chain and PLCgamma2 knockouts with a greater decrease observed in LAT knockouts. Dramatic reduction was observed in wild-types treated with hirudin, with abolished thrombus formation only observed in FcR gamma-chain knockouts treated with hirudin. GPVI-FcR gamma-chain, LAT and PLCgamma2 are essential for thrombus generation and stability in this laser-induced model of injury. More importantly, a greater role for LAT was identified, which may reflect a role for it downstream of a second matrix protein receptor. However, inhibition of platelet activation by matrix proteins and thrombin generation are both required to maximally prevent thrombus formation.

  7. Use of a Fibrinogen/Thrombin-Based Collagen Fleece (TachoComb, TachoSil) With a Stapled Closure to Prevent Pancreatic Fistula Formation Following Distal Pancreatectomy.

    Science.gov (United States)

    Mita, Kazuhito; Ito, Hideto; Murabayashi, Ryo; Asakawa, Hideki; Nabetani, Masashi; Kamasako, Akira; Koizumi, Kazuya; Hayashi, Takashi

    2015-12-01

    Postoperative pancreatic fistula formation remains a source of significant morbidity following distal pancreatectomy. The aim of this study was to evaluate the rate of clinically significant fistulas (International Study Group on Pancreatic Fistula grade B and grade C) after distal pancreatectomy using a fibrinogen/thrombin-based collagen fleece (TachoComb, TachoSil) with a stapled closure. Seventy-five patients underwent distal pancreatectomy at our institution between January 2005 and March 2014. A fibrinogen/thrombin-based collagen fleece was applied to the staple line of the pancreas before stapling. Twenty-six patients (34.7%) developed a pancreatic fistula, 8 patients (10.7%) developed a grade B fistula, and no patients developed a grade C fistula. The duration of the drain was significantly different in patients with or without a pancreatic fistula (8.0 ± 4.5 vs. 5.4 ± 1.3 days, P = .0003). Histological analysis showed that there was a tight covering with the fibrinogen/thrombin-based collagen fleece. The fibrinogen/thrombin-based collagen fleece (TachoComb, TachoSil) with a stapled closure has low rates of fistula formation and provides a safe alternative to the conventional stapled technique in distal pancreatectomy. © The Author(s) 2015.

  8. The Anopheles gambiae cE5, a tight- and fast-binding thrombin inhibitor with post-transcriptionally regulated salivary-restricted expression

    Czech Academy of Sciences Publication Activity Database

    Ronca, R.; Kotsyfakis, Michalis; Lombardo, F.; Rizzo, C.; Currà, C.; Ponzi, M.; Fiorentino, G.; Ribeiro, J.M.C.; Arcà, B.

    2012-01-01

    Roč. 42, č. 9 (2012), s. 610-620 ISSN 0965-1748 R&D Projects: GA ČR GAP502/12/2409 Institutional research plan: CEZ:AV0Z60220518 Keywords : Anopheles * Salivary protein * Anti-thrombin * Anophelin * Hematophagy * Post-transcriptional regulation Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.234, year: 2012

  9. Binding of Thrombin-Activated Platelets to a Fibrin Scaffold through αIIbβ3 Evokes Phosphatidylserine Exposure on Their Cell Surface

    Science.gov (United States)

    Brzoska, Tomasz; Suzuki, Yuko; Mogami, Hideo; Sano, Hideto; Urano, Tetsumei

    2013-01-01

    Recently, by employing intra-vital confocal microscopy, we demonstrated that platelets expose phosphatidylserine (PS) and fibrin accumulate only in the center of the thrombus but not in its periphery. To address the question how exposure of platelet anionic phospholipids is regulated within the thrombus, an in-vitro experiment using diluted platelet-rich plasma was employed, in which the fibrin network was formed in the presence of platelets, and PS exposure on the platelet surface was analyzed using Confocal Laser Scanning Microscopy. Almost all platelets exposed PS after treatment with tissue factor, thrombin or ionomycin. Argatroban abrogated fibrin network formation in all samples, however, platelet PS exposure was inhibited only in tissue factor- and thrombin-treated samples but not in ionomycin-treated samples. FK633, an αIIbβ3 antagonist, and cytochalasin B impaired platelet binding to the fibrin scaffold and significantly reduced PS exposure evoked by thrombin. Gly-Pro-Arg-Pro amide abrogated not only fibrin network formation, but also PS exposure on platelets without suppressing platelet binding to fibrin/fibrinogen. These results suggest that outside-in signals in platelets generated by their binding to the rigid fibrin network are essential for PS exposure after thrombin treatment. PMID:23383331

  10. Binding of thrombin-activated platelets to a fibrin scaffold through α(IIb)β₃ evokes phosphatidylserine exposure on their cell surface.

    Science.gov (United States)

    Brzoska, Tomasz; Suzuki, Yuko; Mogami, Hideo; Sano, Hideto; Urano, Tetsumei

    2013-01-01

    Recently, by employing intra-vital confocal microscopy, we demonstrated that platelets expose phosphatidylserine (PS) and fibrin accumulate only in the center of the thrombus but not in its periphery. To address the question how exposure of platelet anionic phospholipids is regulated within the thrombus, an in-vitro experiment using diluted platelet-rich plasma was employed, in which the fibrin network was formed in the presence of platelets, and PS exposure on the platelet surface was analyzed using Confocal Laser Scanning Microscopy. Almost all platelets exposed PS after treatment with tissue factor, thrombin or ionomycin. Argatroban abrogated fibrin network formation in all samples, however, platelet PS exposure was inhibited only in tissue factor- and thrombin-treated samples but not in ionomycin-treated samples. FK633, an α(IIb)β₃ antagonist, and cytochalasin B impaired platelet binding to the fibrin scaffold and significantly reduced PS exposure evoked by thrombin. Gly-Pro-Arg-Pro amide abrogated not only fibrin network formation, but also PS exposure on platelets without suppressing platelet binding to fibrin/fibrinogen. These results suggest that outside-in signals in platelets generated by their binding to the rigid fibrin network are essential for PS exposure after thrombin treatment.

  11. Binding of thrombin-activated platelets to a fibrin scaffold through α(IIbβ₃ evokes phosphatidylserine exposure on their cell surface.

    Directory of Open Access Journals (Sweden)

    Tomasz Brzoska

    Full Text Available Recently, by employing intra-vital confocal microscopy, we demonstrated that platelets expose phosphatidylserine (PS and fibrin accumulate only in the center of the thrombus but not in its periphery. To address the question how exposure of platelet anionic phospholipids is regulated within the thrombus, an in-vitro experiment using diluted platelet-rich plasma was employed, in which the fibrin network was formed in the presence of platelets, and PS exposure on the platelet surface was analyzed using Confocal Laser Scanning Microscopy. Almost all platelets exposed PS after treatment with tissue factor, thrombin or ionomycin. Argatroban abrogated fibrin network formation in all samples, however, platelet PS exposure was inhibited only in tissue factor- and thrombin-treated samples but not in ionomycin-treated samples. FK633, an α(IIbβ₃ antagonist, and cytochalasin B impaired platelet binding to the fibrin scaffold and significantly reduced PS exposure evoked by thrombin. Gly-Pro-Arg-Pro amide abrogated not only fibrin network formation, but also PS exposure on platelets without suppressing platelet binding to fibrin/fibrinogen. These results suggest that outside-in signals in platelets generated by their binding to the rigid fibrin network are essential for PS exposure after thrombin treatment.

  12. Patients with deep venous thrombosis and thrombophilia risk factors have a specific prolongation of the lag time in a chromogenic thrombin generation assay

    NARCIS (Netherlands)

    Haas, F.J.L.M.; Kluft, C.; Biesma, D.H.; Schutgens, R.E.G.

    2011-01-01

    The objective of the present study was to evaluate the influence of thrombophilia risk factors on variables of a chromogenic thrombin generation assay (ETP) in a setting with acute deep venous thrombosis (DVT) and non-DVT patients. In 152 outpatients suspected for DVT, the results of thrombophilia

  13. Optimizing electrode-attached redox-peptide systems for kinetic characterization of protease action on immobilized substrates. Observation of dissimilar behavior of trypsin and thrombin enzymes.

    Science.gov (United States)

    Anne, Agnès; Chovin, Arnaud; Demaille, Christophe

    2012-06-12

    In this work, we experimentally address the issue of optimizing gold electrode attached ferrocene (Fc)-peptide systems for kinetic measurements of protease action. Considering human α-thrombin and bovine trypsin as proteases of interest, we show that the recurring problem of incomplete cleavage of the peptide layer by these enzymes can be solved by using ultraflat template-stripped gold, instead of polished polycrystalline gold, as the Fc-peptide bearing electrode material. We describe how these fragile surfaces can be mounted in a rotating disk configuration so that enzyme mass transfer no longer limits the overall measured cleavage kinetics. Finally, we demonstrate that, once the system has been optimized, in situ real-time cyclic voltammetry monitoring of the protease action can yield high-quality kinetic data, showing no sign of interfering effects. The cleavage progress curves then closely match the Langmuirian variation expected for a kinetically controlled surface process. Global fit of the progress curves yield accurate values of the peptide cleavage rate for both trypsin and thrombin. It is shown that, whereas trypsin action on the surface-attached peptide closely follows Michaelis-Menten kinetics, thrombin displays a specific and unexpected behavior characterized by a nearly enzyme-concentration-independent cleavage rate in the subnanomolar enzyme concentration range. The reason for this behavior has still to be clarified, but its occurrence may limit the sensitivity of thrombin sensors based on Fc-peptide layers.

  14. Duplex/quadruplex oligonucleotides: Role of the duplex domain in the stabilization of a new generation of highly effective anti-thrombin aptamers.

    Science.gov (United States)

    Russo Krauss, Irene; Napolitano, Valeria; Petraccone, Luigi; Troisi, Romualdo; Spiridonova, Vera; Mattia, Carlo Andrea; Sica, Filomena

    2018-02-01

    Recently, mixed duplex/quadruplex oligonucleotides have attracted great interest for use as biomedical aptamers. In the case of anti-thrombin aptamers, the addition of duplex-forming sequences to a G-quadruplex module identical or very similar to the best-known G-quadruplex of the Thrombin Binding Aptamer (HD1) results in new or improved biological properties, such as higher activity or different recognition properties with respect to HD1. Remarkably, this bimodular fold was hypothesized, based on its sequence, for the only anti-thrombin aptamer in advanced clinical trial, NU172. Whereas cation modulation of G-quadruplex conformation and stability is well characterized, only few data from similar analysis on duplex/quadruplex oligonucleotides exist. Here we have performed a characterization of structure and stability of four different duplex/quadruplex anti-thrombin aptamers, including NU172, in the presence of different cations and in physiological-mimicking conditions in comparison to HD1, by means of spectroscopic techniques (UV and circular dichroism) and differential scanning calorimetry. Our data show a strong reciprocal influence of each domain on the stability of the other and in particular suggest a stabilizing effect of the duplex region in the presence of solutions mimicking the physiological conditions, strengthening the idea that bimodular aptamers present better therapeutic potentialities than those containing a single G-quadruplex domain. Copyright © 2017 Elsevier B.V. All rights reserved.

  15. LANL MTI calibration team experience

    Science.gov (United States)

    Bender, Steven C.; Atkins, William H.; Clodius, William B.; Little, Cynthia K.; Christensen, R. Wynn

    2004-01-01

    The Multispectral Thermal Imager (MTI) was designed as an imaging radiometer with absolute calibration requirements established by Department of Energy (DOE) mission goals. Particular emphasis was given to water surface temperature retrieval using two mid wave and three long wave infrared spectral bands, the fundamental requirement was a surface temperature determination of 1K at the 68% confidence level. For the ten solar reflective bands a one-sigma radiometric performance goal of 3% was established. In order to address these technical challenges a calibration facility was constructed containing newly designed sources that were calibrated at NIST. Additionally, the design of the payload and its onboard calibration system supported post launch maintenance and update of the ground calibration. The on-orbit calibration philosophy also included vicarious techniques using ocean buoys, playas and other instrumented sites; these became increasingly important subsequent to an electrical failure which disabled the onboard calibration system. This paper offers various relevant lessons learned in the eight-year process of reducing to practice the calibration capability required by the scientific mission. The discussion presented will include observations pertinent to operational and procedural issues as well as hardware experiences; the validity of some of the initial assumptions will also be explored.

  16. Field calibration of cup anemometers

    DEFF Research Database (Denmark)

    Kristensen, L.; Jensen, G.; Hansen, A.

    2001-01-01

    An outdoor calibration facility for cup anemometers, where the signals from 10 anemometers of which at least one is a reference can be can be recorded simultaneously, has been established. The results are discussed with special emphasis on the statisticalsignificance of the calibration expressions...

  17. Cobalt source calibration

    International Nuclear Information System (INIS)

    Rizvi, H.M.

    1999-01-01

    The data obtained from these tests determine the dose rate of the two cobalt sources in SRTC. Building 774-A houses one of these sources while the other resides in room C-067 of Building 773-A. The data from this experiment shows the following: (1) The dose rate of the No.2 cobalt source in Building 774-A measured 1.073 x 10 5 rad/h (June 17, 1999). The dose rate of the Shepherd Model 109 Gamma cobalt source in Building 773-A measured 9.27 x 10 5 rad/h (June 25, 1999). These rates come from placing the graduated cylinder containing the dosimeter solution in the center of the irradiation chamber. (2) Two calibration tests in the 774-A source placed the graduated cylinder with the dosimeter solution approximately 1.5 inches off center in the axial direction. This movement of the sample reduced the measured dose rate 0.92% from 1.083 x 10 5 rad/h to 1.073 x 10 5 rad/h. and (3) A similar test in the cobalt source in 773-A placed the graduated cylinder approximately 2.0 inches off center in the axial direction. This change in position reduced the measured dose rate by 10.34% from 1.036 x 10 6 to 9.27 x 10 5 . This testing used chemical dosimetry to measure the dose rate of a radioactive source. In this method, one determines the dose by the chemical change that takes place in the dosimeter. For this calibration experiment, the author used a Fricke (ferrous ammonium sulfate) dosimeter. This solution works well for dose rates to 10 7 rad/h. During irradiation of the Fricke dosimeter solution the Fe 2+ ions ionize to Fe 3+ . When this occurs, the solution acquires a slightly darker tint (not visible to the human eye). To determine the magnitude of the change in Fe ions, one places the solution in an UV-VIS Spectrophotometer. The UV-VIS Spectrophotometer measures the absorbency of the solution. Dividing the absorbency by the total time (in minutes) of exposure yields the dose rate

  18. AUTOMATION OF IMAGE DATA PROCESSING

    Directory of Open Access Journals (Sweden)

    Preuss Ryszard

    2014-12-01

    Full Text Available This article discusses the current capabilities of automate processing of the image data on the example of using PhotoScan software by Agisoft . At present, image data obtained by various registration systems (metric and non - metric cameras placed on airplanes , satellites , or more often on UAVs is used to create photogrammetric products. Multiple registrations of object or land area (large groups of photos are captured are usually performed in order to eliminate obscured area as well as to raise the final accuracy of the photogrammetric product. Because of such a situation t he geometry of the resulting image blocks is far from the typical configuration of images . For fast images georeferencing automatic image matching algorithms are currently applied . They can create a model of a block in the local coordinate system or using initial exterior orientation and measured control points can provide image georeference in an external reference frame. In the case of non - metric image application, it is also possible to carry out self - calibration process at this stage . Image matching algorithm is also used in generation of dense point clouds reconstructing spatial shape of the object ( area. In subsequent processing steps it is possible to obtain typical photogrammetric products such as orthomosaic , DSM or DTM and a photorealistic solid model of an object . All aforementioned processing steps are implemented in a single program in contrary to standard commercial software dividing all steps into dedicated modules . I mage processing leading to final geo referenced products can be fully automated including sequential implementation of the processing steps at predetermined control parameters . The paper presents the practical results of the application fully automatic generation of othomosaic for both images obtained by a metric Vexell camera and a block of images acquired by a non - metric UAV system.

  19. Systematic review automation technologies

    Science.gov (United States)

    2014-01-01

    Systematic reviews, a cornerstone of evidence-based medicine, are not produced quickly enough to support clinical practice. The cost of production, availability of the requisite expertise and timeliness are often quoted as major contributors for the delay. This detailed survey of the state of the art of information systems designed to support or automate individual tasks in the systematic review, and in particular systematic reviews of randomized controlled clinical trials, reveals trends that see the convergence of several parallel research projects. We surveyed literature describing informatics systems that support or automate the processes of systematic review or each of the tasks of the systematic review. Several projects focus on automating, simplifying and/or streamlining specific tasks of the systematic review. Some tasks are already fully automated while others are still largely manual. In this review, we describe each task and the effect that its automation would have on the entire systematic review process, summarize the existing information system support for each task, and highlight where further research is needed for realizing automation for the task. Integration of the systems that automate systematic review tasks may lead to a revised systematic review workflow. We envisage the optimized workflow will lead to system in which each systematic review is described as a computer program that automatically retrieves relevant trials, appraises them, extracts and synthesizes data, evaluates the risk of bias, performs meta-analysis calculations, and produces a report in real time. PMID:25005128

  20. Liquid Krypton Calorimeter Calibration Software

    CERN Document Server

    Hughes, Christina Lindsay

    2013-01-01

    Calibration of the liquid krypton calorimeter (LKr) of the NA62 experiment is managed by a set of standalone programs, or an online calibration driver. These programs are similar to those used by NA48, but have been updated to utilize classes and translated to C++ while maintaining a common functionality. A set of classes developed to handle communication with hardware was used to develop the three standalone programs as well as the main driver program for online calibration between bursts. The main calibration driver has been designed to respond to run control commands and receive burst data, both transmitted via DIM. In order to facilitate the process of reading in calibration parameters, a serializable class has been introduced, allowing the replacement of standard text files with XML configuration files.