text:
|Publication number
|Publication type
|Application number
|Publication date
|Jul 3, 2001
|Filing date
|Nov 5, 1999
|Priority date
|Nov 5, 1999
|Fee status
|Also published as
|Publication number
|Philip Pivarnik, He Cao, Stephen V. Letcher, A. Garth Rand
|Original Assignee
|The Board Of Governors For Higher Education, State Of Rhode Island And Providence Plantations
|Export Citation
|BiBTeX, EndNote, RefMan
|Patent Citations (30), Non-Patent Citations (10), Referenced by (55), Classifications (53), Legal Events (8)
|External Links: USPTO, USPTO Assignment, Espacenet
The U.S. Government has a paid up license in this invention and the right in limited circumstances to require the patent owner to license others on reasonable terms as provided for by the terms of Grant No. 97-35201-4480 from the US Department of Agricultural National Research Initiative Competitive Grant Program on Food Safety and Contract No. DAAK6095C2035 from the US Army Soldier Systems Command/Natick.
1. Field of the Invention
A fiber-optic biosensor for assaying an analyte of interest.
2. Description of the Related Art
Concern over virulent pathogens contaminating the food supply has the potential to put large segments of the population at risk. The scientific estimates project that between 24 million and 81 million people become ill from foodborne diarrheal disease each year in the United States. The impact of this food contamination costs between $5 billion and $17 billion in medical care costs and lost productivity. Most toxicologists and food scientists agree these figures probably represent about 75% of the whole food safety risks, confirming that microbial pathogens are considered a serious hazard.
The impact due to Salmonella has been estimated at >$4 billion from more than 3 million cases/year. The lack of success in effectively controlling the growth of these pathogens in foods suggests that the best approach would be early and rapid detection. Approximately 5 million analytical tests are performed annually in the U.S., which makes detection a high priority for diagnostic technology.
Traditional methods to identify and quantitate contaminants in foods include physicochemical, biological and serological tests. Most of these approaches lack sufficient sensitivity, selectivity, and take days to perform. There have been many attempts to provide faster and convenient detection methods. Most of these detection systems, although referred to as "rapid methods", still rely on culturing procedures to selectively amplify microbial populations.
The development of food immunoassays to improve this process has provided increased speed, simplicity, and effectiveness. However, these current procedures are expensive, usually require an enrichment/concentration step and still require hours to days for a final result. The adaptation of this immunoassay technology to biosensors has the potential to take immunoassays into the realm of rapid and reusable biosensors. The application of biosensors to the detection of Salmonella has the potential to contribute directly to the production and processing of safer and healthier food.
Biosensors are analytical devices which incorporate biologically active material in intimate contact with a transduction element to selectively and repeatedly detect analytes in products like food and food raw materials. The effective development of biosensors has become multidisciplinary, relying on biochemistry and biotechnology to provide the sensing elements through immobilization techniques and membrane technology, but also requires expertise in microelectronics, optical, acoustical and advanced signal processing. This dramatic combination can be effectively applied in the area of food safety, for the elimination of serious health risks by rapid detection of specific pathogens.
A compact fiber-optic evanescent-wave biosensor system which features an all-fiber optical design and red semiconductor-laser excitation has been developed. C. Zhou, P. Pivarnik, S. Auger, A Rand and S. Letcher, "A compact fiber optic immunosensor for Salmonella based on evanescent wave excitation", Sensors and Actuators B 42, pp. 169-175, 1997. Tapered fiber tips with immobilized antibodies for Salmonella attached were studied in different shapes and treatments and optimized. The system response for Salmonella concentration was established and could determine as low as 104 colony forming units (CFU/mL) in 1 hour with a sandwich immunoassay format. Acoustic enhancement of the fiber-optic biosensor with ultrasonic manipulation of suspended particles has been shown to be an effective way to increase the sensitivity. C. Zhou, P Pivarnik, A. Rand, and S. Letcher, "Acoustic standing-wave enhancement of a fiber optic Salmonella biosensor", Biosensors and Bioelectronics 13, pp 495-500, 1998. Polystyrene microspheres (6-mm diameter) coated with immobilized antibodies were allowed to capture the antigens (Salmonella), which in turn captured antibodies labeled with fluorescent dye molecules. Then the entire structure was moved to the center of the acoustic cell, where the optical fiber with its cladding removed was located. The fluorescent signal was greatly increased over the signal without acoustic positioning. Multiple use of the system was also rapid because the fiber tip could be reused indefinitely (no antibodies were attached)—the flow cell just needed to be flushed out with a buffer solution and was ready for reuse.
Although the use of evanescent wave and tapered fibers provided average sensitivity and measurement potential, these systems did not achieve rapid and direct pathogen detection for food use. Currently the best available commercial system has a sensitivity of 104 Colony Forming Units/ml (CFU/ml) and measurement potential of 107 to 108 CFU/ml . The time to complete an assay is usually 3 to 4 hours.
The present invention comprises a fiber optic biosensor and method for the detection of pathogens, i.e. bacteria, viruses and/or toxins. The fiber-optic biosensor comprises a laser in communication with an excitation fiber. The excitation fiber is in communication with a collection fiber and the excitation and collection fibers are in communication with a magnetic focusing probe. The collection fiber is in communication with a transmission fiber which is in communication with means for detecting, collecting and measuring fluorescent signals. The present invention reduces the background fluorescence from unbound antibodies and eliminates several rinsing steps thereby resulting in a simple rapid assay.
Broadly the invention comprises magnetic focusing of paramagnetic microspheres with a fiber optic biosensor. Microspheres with immobilized antibodies interact throughout the analyte containing the target antigens, which, in turn, capture fluorescent-labeled antibodies in a standard sandwich assay. The bound antigen/antibody/fluorescent antibody complexes are magnetically attracted to the magnetic focusing probe of the fiber optic biosensor which contains the sensing volume of the excitation and collection fibers, while the uncaptured labeled fluorescent antibodies remain in bulk solution thereby reducing background fluorescence.
In contrast to the prior art fiber-optic immunoassays that are acoustically focused and have several steps for detection, the present invention provides an immunoassay that uses magnetic focusing and only requires three steps, specifically; 1) immunocapture and labeling, 2) rinsing and 3) focusing and measurement of fluorescent signal. Furthermore, the fiber-optic immunosensor of the present invention is more efficient than prior art fluorescent immunosensors because prior art fluorescent immunosensors do not concentrate the bound antigen/antibody/fluorescent antibody complexes into the optically active region (field of view) of the fiber.
In a preferred embodiment of the invention a primary antibody specific for the antigen (pathogen) to be detected is coated on a magnetic bead and a secondary antibody is conjugated to a marker. A food sample is prepared and added to the medium. If the expected pathogen, such as Salmonella Typhimurium is present, the pathogen binds to both antibodies forming a magnetic complex. The magnetic complex is attracted to the magnetic focusing probe of the fiber-optic immunosensor containing the excitation and collecting fibers. The fibers are attached to the transmission fiber. The transmission fiber transmits the fluorescent signal received from the contacting fiber to means for detecting, collecting, and measuring the fluorescent signal. The means for detecting, collecting and measuring the fluorescent signal can be a fiber optic spectrometer in communication with a computer or a PIN detector in communication with an optical power meter.
In another aspect of the invention, a sample from the blood of mammals, fish or fowl is prepared and added to a medium comprised of primary antibodies specific for the anitgens (pathogens) to be detected whereby the primary antibodies are coated on a magnetic bead and a secondary antibody conjugated to a marker. The fiber-optic immunosensor of the present invention could then be utilized as described above.
In yet another aspect of the invention, the fiber-optic immunosensor is a portable unit that can be used for field testing.
FIG. 1 is a schematic of a fiber optic immunosensor embodying the invention;
FIG. 2 is a schematic view of an embodiment of the fiber optic magnetic focusing probe of the invention;
FIG. 3 is a perspective view of an embodiment of the magnetic focusing probe with parallel fibers;
FIG. 4 is a diagram of an increase of fluorescent signals during focusing;
FIG. 5 is a chart of the effect of a cuvette wall thickness on the fluorescent signal;
FIG. 6 is a chart on the reproducibility of the focusing step; and
FIG. 7 is a diagram for the response of magnetically focused assay to increasing concentrations of Salmonella typhimurium.
Referring to FIG. 1, a laser 10 having a 650 nm, 5-mW diode laser (Power Technology Corp.) is in communication with a line filter 12 (Omega Optical) which has its transmission maximum at 654 nm. The line filter 12 is in communication with a collimator 14 which is in communication with an exciting fiber 16. The exciting fiber 16 is in communication with a fiber optic magnetic focusing probe 18. The fiber magnetic focusing probe 18 is in communication with a collecting fiber 20. The collecting fiber 20 and the exciting fiber 16 are made from 500-micron (480-micron core) polymethyl methacrylate optical grade fibers. (ESKA brand).
The collecting fiber 20 is in communication with a collimator 34 which is in communication with a long pass filer 22. The long pass filter 22 has a cutoff of 670 nm and is made by Omega Optical. The long pass filter 22 is in communication with a transmission fiber 24 via collimator 36. The transmission fiber 24 is made of silica with a 400 micron core.
In the preferred embodiment of the invention the laser 10 is a HeNe laser with a wavelength maximum of 633 nm. The line filter 12 is omitted. The long pass filter 22 has a 50% cutoff of 664 nm. The transmission fiber 24 is in communication with a fiber optic spectrometer 26 and a computer 28. The fiber optic spectrometer 26 is an Ocean Optics (Dunedin, Fla.) model PS1000 portable spectrometer. The computer 28 is an IBM compatible computer which controls the fiber optic spectrometer 26 and collects data with the Spectro Scope� software from Ocean Optics.
In an alternate embodiment the transmission fiber 24 is in communication with a PIN detector 30 and an optical power meter 32. The PIN detector 30 and the optical power meter 32 (Model 835) are made by Newport Corporation. The wavelength range of the CCD array in the spectrometer is 250 nm to 750 nm. The fluorescent signal from the CY5 dye is detected at approximately 674 nm. No interference is observed from the HeNe laser.
Referring to FIGS. 2 and 3, a schematic and perspective view of the fiber optic magnetic focusing probe 18 is shown. The fiber optic magnetic focusing probe 18 comprises a tapered soft-iron tip 40 attached to a rare-earth permanent magnet 42 that is rated at 4600 gauss to form the probe 18. The optical fibers 16 and 20 are epoxied with a resin and secured into slits 44 and 46 that have been cut into the probe 18, 180� apart, see FIG. 3. Each optical fiber 16 and 20 is tilted at 18� from the normal to a wall 8 of a cuvette (not shown) in order to provide maximum overlap of their optical fields at the cuvette wall where the paramagnetic particles gather. The tapered soft-iron tip 40 is ground and polished and the outer edges of the tapered soft-iron tip 40 are slightly beveled so that the collection of paramagnetic particles is centered in front of the fibers. The end of the tip is about 1.2 mm in diameter. The magnet 42 has a diameter of 0.6 cm and a total length of about 3 cm.
Materials, Chemicals, Microorganisms, and Supplies
Salmonella typhirnium strain ATCC 13311 was purchased from the American Type Culture Collection (Rockeville, Md.). Nutrient broth and nutrient agar were obtained from Difco laboratories (Detroit, Mich.) and used for growing and counting the Salmonella. Dynabeads� anti-Salmonella paramagnetic spheres, 2.8 microns in diameter, were purchased from Dnyal, Inc. (Oslo, Norway). Purified (>95% IgG) polyclonal antibodies specific to Salmonella species were obtained from Biodesign International (Kennebunk, Me.). Fluorolink-AB™ Reactive Dye labeling kits from Amersham Life Sciences (Pittsburgh, Pa.) were used for labeling of the polyclonal antibodies. Fisherbrand� semi-micro disposable methacrylate cuvettes were obtained from Fisher Scientific (Springfield, N.J.). All other chemicals were ACS reagent grade. Reverse osmosis/deionized waster was used in the preparation of all solutions.
Culturing and Preparation of the Microorganisms for Assay.
Aliquots of Salmonella typhimurium cultures were stored at −80� C. until needed. Immunoassay cultures were prepared by thawing an aliquot of the organism and inoculating a tube of sterile nutrient broth with 1 to 2 loops of organisms. The freshly inoculated broth was then incubated at 37� C. for 9 hours. The newly grown organisms were then used immediately for the assay or stored at 4� C. until they were used for the immunoassay, usually overnight. An aliquot of every freshly prepared culture was checked with the Gram staining procedure to verify the presence of Salmonella typhimurium and check for contaminants. The cultures were prepared for immunoassay by centrifugation at 8000�g for 15 minutes at 4� C. The spent nutrient broth was decanted and the cells were resuspended in phosphate buffered saline with 0.05% Tween 20 (PBST). An aliquot of the resuspended organisms was transferred into a vial of phosphate buffered saline (PBS) and then serially diluted in PBS for plate counts. The organisms were added to nutrient agar plates and spread on the surface and incubated at 37� C. for 24 hours for plate counts.
Immunoassay Procedures
Initially a two-step assay was run with separate steps for binding to the microspheres and labeling of the microorganisms. The organisms suspended in PBST were added to 20 μl of Dynabeads� anti-Salmonella in a microcentrifuge tube and mixed thoroughly. The mixture was mixed and incubated at room temperature for 30 minutes. After the incubation the tubes were placed into a magnetic separator and the spheres were allowed to collect on the side of the tube for 3 minutes. Then the unbound organisms in the buffer were carefully drawn from the tube, being sure not to disturb the beads collected on the sides of the tube. A 1 ml aliquot of PBST was added to the tube and thoroughly mixed. Then the tubes were placed in the magnetic separator and rinse was repeated. Each tube was rinsed 2 times. Then the spheres, with the salmonella attached were incubated with diluted secondary antibody in PBST for 30 minutes at room temperature with constant mixing. The samples were then rinsed 2 times with PBST. Finally, the microspheres with Salmonella attached were resuspended in 0.5 ml of PBST and placed into a semi-micro cuvette for the focusing assay.
For a one-step procedure the Salmonella, Dynabeads�, and CY5-IgG were combined in the microcentrifuge tube and incubated for 30 minutes at room temperature with constant mixing. Following the one-step binding and labeling procedure the tubes were placed in the magnetic separator and rinsed 2 times with PBST, and then resuspended in 0.5 ml of PBST in a semi-micro cuvette for readings with the magnetic probe. CY5-IgG solutions were diluted from 1:100 to 1:2,000 for the assays.
Results and Discussion
Preparation of secondary antibody. Conjugation of CY5 fluorescent dye with anti-Salmonella IgG. The polyclonal anti-Salmonella IgG was labeled with CY5 using the Fluorolink-Ab™ kits. Four different 1 mg aliquots of the IgG were labeled. The dye/protein ratio of the labeled IgG was calculated from the absorbance reading at 650 nm and 280 nm and the results are presented in FIG. 2. The ratios range from 5.85 to 6.92 and are consistent and with the recommended dye/protein ratios for the procedure, which are 4 to 12. A ratio less than 4 results in too little signal and greater than 12 can result in overlabeling and inactivation of the IgG and/or interference of the signal due to too much dye. A slightly higher ratio may allow for a greater dilution of the secondary antibody and may provide more sensitivity but may not be necessary.
Magnetic Focusing of Immunoassay Samples.
the one-step binding and labeling of the Salmonella procedure was used to prepare samples for the magnetic focusing trials. One-step binding and labeling of the organisms is possible due to the fact that different sources of primary and secondary antibodies were used. the primary antibody was a proprietary antibody attached to the magnetic spheres purchased from Dynal, Inc. (Oslo, Norway) and the secondary antibody purified IgG labeled with CY5 as described above. Results of the three different magnetic focusing trials are presented in FIG. 4. For trials 1 and 2 the dilution of secondary antibody was 1:500 and for Trial 3 the dilution of secondary antibody was 1:100, thus the higher signal from Trial 3 was primarily due to the difference in secondary antibody concentration. The cell concentration for Trial 1 was 2.09�107 CFU/ml and for Trials 2 and 3 was 1.6�107 CFU/ml.
The increase in signal over time demonstrates that the beads were focused in front of the magnetic fiber optic probe to enhance the detection of the organisms by separating the beads from the solution. The curve shows almost no increase in signal in first 30 seconds, followed by the most rapid increase in signal between 30 and 90 seconds, and finally a leveling off of signal at 120 seconds. No substantial increase in sensing occurred between 120 and 180 seconds. At 180 seconds the signal has reached its maximum and continued monitoring of the sample resulted only in a slight decrease as the time is extended. This may be due to bleaching of the dye due to the constant exposure to the strong laser light. Visual inspection of the cuvette after the readings are completed verified that the solution was cleared of magnetic spheres. The spheres had collected in a small, solid circle on the wall of the cuvette in front of the magnetic probe and fibers. The diameter of the circle was 3 to 5 mm.
Several factors affected the time required for focusing and the pattern and size of the beads focused on the side of the cuvette. First, the strength of the magnetic field was the most important factor in determining the time required to focus the sample. Stronger magnets resulted in shorter focusing times and complete removal of the spheres from the solutions. When weaker magnets were used the field was not strong enough to clear all of the paramagnetic spheres from solution. The strength of the magnetic field was inversely proportional to the size of the iron probe used to construct the probe. The volume and dimensions of the cuvette were also critical to the focusing step. Using the semimicro cuvettes the probe could be positioned on the side of the cuvette with the path of the cuvette no more than 5 to 6 mm from the tip of the probe. When the probe is placed in the cuvette window that allows for a 1 cm path length those spheres that are more than 5 mm to 6 mm away require much more time to focus. The end probe tip must be small to ensure that the spheres collect in front of excitation and emission fibers and form a solid, reasonably uniform layer on the side of the cuvette. The force on a paramagnet microsphere is proportional to the strength of the magnetic field and to its spatial gradient. The particles are, therefore, most strongly attracted toward the sharp corners of the magnet. When the large end of a cylindrical magnet is used, the spheres often form a ring around the outside of the magnet resulting in a large opening near the center of the magnet where the excitation and emission fibers must be located.
At 0 minutes the signal from the unfocused spheres was in the range of 9 to 12 picowatts. After the magnetic spheres were focused in front of the fibers the signal increased at least 10 fold (500� dilution of secondary IgG) to 20 fold (100� dilution of secondary IgG). Ultimately, the focusing provides a substantial advantage over reading the signal from the beads suspended in bulk solution. It also eliminates interference from the fluorescence of the unbound secondary antibody in the bulk solution.
Disposable semi-micro cuvettes were used. The thickness of the cuvette cell wall 40 was measured and it was found that for Case A the thickness was 0.90�0.02 mm and for Case B the thickness was 0.80�0.02 mm. The cuvettes were then placed in the sample holder and the signal was recorded for each case and is presented in FIG. 5. For sample 1A, the signal for case B was 52% greater than the signal for case A and for sample 1B the increase in signal was 47%. The thickness of the cuvette wall and the distance of the fibers from the paramagnetic spheres are important in obtaining consistent results.
To test for the reproducibility of the focusing step one sample from each of the trials was inserted into the holder, focused and measured, and then removed from the holder and mixed., and then reinserted into the holder. This was repeated up to 20 times for each sample chosen. After 10 of the readings the fluorescent signal began to decrease and this was attributed to bleaching of the fluorescent dye by the laser. Consequently only data from the first 10 readings was considered valuable. FIG. 6 is a graph of the results. The difference in the absolute signal was attributed to the fact that the secondary antibody was diluted 500 fold in trials 1 and 2 and diluted 100 fold in trial 3. The error bars on the graph are the standard deviations for each sample. The % error, or coefficient of variation, for the samples was 8.0% 5.5%, and 10.6% for Trial 1, Trial 2, and Trial 3, respectively.
Variability, as measured by the coefficient of variation, due solely to the focusing step was less than 11%. An 11% error from the focusing step must be reduced and it is expected that improvements in the positioning of the cuvette, and construction of the probe are expected to decrease this error.
Detection of Salmonella typhimurium with magnetically focused beads was determined by assaying a series of samples with increasing Salmonella concentrations. Three different experiments were run with increasing concentrations of Salmonella. The assay results showed that Salmonella was detected at 105 CFU/ml of sample. The curve form one of these experiments is show in FIG. 7. The data show that with increasing Salmonella concentration the fluorescent signal begins to rise above the baseline at 3.17�105 CFU/ml and increases approximately 1.16�107 CFU/ml, where it reaches a maximum and levels off.
A two-step focusing procedure was developed to decrease analysis time. The advantage of the two-step focusing using a stronger magnet to focus the spheres on to the side of the cuvette wall prior to insertion of the curvette into the holder in front of the probe. The use of a stronger, larger magnet in the first step can reduce the total focusing time to one minute or less. The data show no difference in the quantitative results that could be attributed to the one and two-step procedures. However, one difference between one and two-step focusing is that the spot that the magnetic spheres form in front of the fiber is smaller with the two-step focusing. The two-step procedure can cut the time required to get a fluorescence reading in half.
Magnetic focusing of the spheres in front of the excitation and emission optical fibers increased the fluorescent signal several fold over the signal from the beads uniformly distributed in the bulk solution. The system proved to be very sensitive to the distance from the fibers to the layer of spheres. The process leads to an easy one or two-step focusing that will greatly simplify the measurement of Salmonella. Using the magnetic microspheres to capture the Salmonella increases the interaction of the solid phase immunoassay with the target organism. This leads to a more sensitive assay once the optimum immunoassay parameters have been defined.
Another advantage of using the magnetic focusing is that when the spheres collect in front of the fiber they form an opaque non transparent spot on the side of the cuvette. As the spheres are focused the background signal from the unbound CY5-IgG may be eliminated, or reduced to insignificant levels. This results in a very simple assay that utilizes a one-step binding and labeling step along with a one-step focusing and measurement which is a significant advancement over current procedures that require multiple incubation and rinsing steps. All the reagents can be mixed in a cuvette, or other custom designed sample holder, allowed to incubate for a specified period of time, and then measured by simply inserting the sample holder into a chamber in front of the magnetic probe.
The foregoing description has been limited to a specific embodiment of the invention. It will be apparent, however, that variations and modifications can be made to the invention, with the attainment of some or all of the advantages of the invention. Therefore, it is the object of the appended claims to cover all such variations and modifications as come within the true spirit and scope of the invention.
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|Dec 6, 2016
|Becton, Dickinson And Company
|Devices and methods for manipulating components in a fluid sample
|Jan 31, 2001
|Oct 31, 2002
|Mcdevitt John T.
|Magnetic-based placement and retention of sensor elements in a sensor array
|Feb 5, 2002
|Jan 2, 2003
|Willson C. Grant
|Use of mesoscale self-assembly and recognition to effect delivery of sensing reagent for arrayed sensors
|Jan 31, 2001
|Apr 3, 2003
|Mcdevitt John T.
|Method and system for collecting and transmitting chemical information
|May 10, 2001
|Sep 25, 2003
|Kazuhiro Niwa
|Chemical sensor device
|Jan 31, 2001
|Oct 2, 2003
|Mcdevitt John T.
|Portable sensor array system
|Nov 12, 2003
|May 20, 2004
|Matsushita Electric Industrial Co., Ltd.
|Specific coupling reaction measuring method and reagent kit and specific coupling reaction measuring apparatus for use in the same
|May 17, 2004
|Jan 27, 2005
|Meiring Jason E.
|Image and part recognition technology
|Nov 4, 2004
|Nov 17, 2005
|Fortebio, Inc.
|Fiber-optic assay apparatus based on phase-shift interferometry
|Jan 6, 2006
|Jul 13, 2006
|Fortebio, Inc.
|Enzyme activity measurement using bio-layer interferometry
|Aug 21, 2006
|Dec 14, 2006
|Matsushita Electric Industrial Co., Ltd.
|Specific coupling reaction measuring method and reagent kit and specific coupling reaction measuring apparatus for use in the same
|Aug 7, 2006
|Feb 7, 2008
|Hardman Clayton M
|System and method for detecting specific binding reactions using magnetic labels
|Dec 14, 2007
|Jun 19, 2008
|Fortebio, Inc.
|Fiber-Optic Assay Apparatus Based on Phase-Shift Interferometry
|Apr 8, 2008
|Aug 7, 2008
|Fortebio, Inc.
|Fiber-optic assay apparatus based on phase-shift interferometry
|Jul 16, 2008
|Nov 6, 2008
|Board Of Regents, The University Of Texas System
|Image And Part Recognition Technology
|Sep 11, 2008
|Mar 12, 2009
|Baxter International Inc.
|Infusion therapy sensor system
|Dec 15, 2008
|Dec 31, 2009
|Hrair Kirakossian
|Rapid particle detection assay
|Sep 14, 2007
|Apr 15, 2010
|Fortebio, Inc.
|Amine-Reactive Biosensor
|Mar 13, 2013
|Aug 1, 2013
|Hewlett-Packard Development Company, L.P.
|Electrically driven devices for surface enhanced raman spectroscopy
|Mar 15, 2013
|Jun 9, 2015
|Becton, Dickinson And Company
|Vessel assembly
|Jan 29, 2015
|Dec 6, 2016
|Becton, Dickinson And Company
|May 29, 2015
|Aug 26, 2015
|Portable quick detection method for pathogenic bacteria
|Apr 19, 2007
|Oct 22, 2008
|Hach Lange GmbH
|Method for determining the concentration of analytes
|Apr 15, 2008
|Oct 22, 2008
|Hach Lange GmbH
|Method for determining the concentration of analytes
|Jul 24, 2007
|Apr 8, 2009
|Becton, Dickinson & Company
|Assay particle concentration and imaging apparatus and method
|Jul 24, 2007
|Nov 10, 2010
|Becton Dickinson Co
|Assay particle concentration and imaging apparatus and method
|Jan 9, 2009
|Jul 15, 2009
|Becton, Dickinson and Company
|Rapid particle detection assay
|Jan 9, 2009
|Mar 24, 2010
|Becton, Dickinson and Company
|Rapid particle detection assay
|Jul 24, 2007
|Jul 31, 2013
|Becton, Dickinson & Company
|Assay particle concentration and imaging apparatus and method
|Jan 31, 2002
|Dec 27, 2002
|Board Of Regents, The University Of Texas System
|Magnetic-based placement and retention of sensor elements in a sensor array
|Jan 31, 2002
|Jun 5, 2003
|Univ Texas
|Magnetic-based placement and retention of sensor elements in a sensor array
|Mar 30, 2012
|Oct 4, 2012
|Qinetiq Limited
|U.S. Classification
|422/82.07, 435/290.1, 436/526, 356/244, 435/808, 436/532, 436/534, 436/166, 436/806, 435/7.95, 436/805, 435/288.7, 436/501, 435/7.93, 436/533, 436/172, 356/303, 436/73, 422/50, 435/7.1, 435/287.2, 356/300, 356/326, 422/82.05, 422/52, 436/85, 356/301, 435/287.1, 436/164, 435/7.5, 422/82.08, 436/530, 436/800, 356/311, 436/86, 356/319, 435/289.1, 435/7.94, 436/63, 436/74, 436/546, 422/504
|International Classification
|Cooperative Classification
|European Classification
|Jan 31, 2000
Owner name: BOARD OF GOVERNORS FOR HIGHER EDUCATION, STATE OF
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|Dec 23, 2004
|Fee payment
Year of fee payment: 4
|Jan 13, 2009
|Maintenance fee reminder mailed
|Apr 6, 2009
|Fee payment
Year of fee payment: 8
|Apr 6, 2009
|Surcharge for late payment
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Owner name: RHODE ISLAND BOARD OF EDUCATION, STATE OF RHODE IS
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|Jun 21, 2017
Owner name: COUNCIL ON POSTSECONDARY EDUCATION, RHODE ISLAND
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Enjoying your computer projects and the work which you are working on can be hard to do. This is when you should know more information about why you should be using the CNC - PartMaster CNC File to help you in getting all the files and parts made in a timely manner. Without knowing about the CNC - PartMaster CNC File you will easily get swept up in thinking that you can get the work done, but in fact it will take you more time while your competitors are getting the information out to the public and starting to make money off of the idea you had, but did not have the proper file. With the CNC - PartMaster CNC File you will not be concerned about this because you will have the files available to you for using and know it will get your computer work done right.
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Dolphin Cad Cam PartMaster Having the ability to machine your own parts can be a good thing, but not everyone is going to have this option. This is when you should know...
User Reviews

subset: commoncrawl


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Travel, poetry, law, interior design, community work and business.
Family, education and the development of young children are my true loves. Besides running my kindergarten which will be 20 years old in 2013, I also do some corporate work on a part-time basis. This work is business and corporate relations development with project procurement. I find this challenging and enjoy it very much. I am blessed with 6 adult children who are successful in their own right.
Smile even in the hardest of times. God willing from every unfortunate experience we have, is born something new and exciting that will enhance our wisdom for future generations ~


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News : TeleVoice Moves Headquarters to New Offices
Houston, TX, USA, Aug 31, 2016 -- To accommodate its continued growth, TeleVoice, a provider of customized call center solutions, has moved to new offices that are substantially larger than its previous location.
The offices are still in Houston and feature an audio recording studio for production of IVR prompts using the company's experienced voice talent team. Since each client has unique criteria for its interactive voice response (IVR) application, this in-house studio allows TeleVoice to be in complete control of the quality of the recordings and facilitates faster turnaround for time-sensitive projects.
The new facilities also include an expanded test lab allowing for simultaneous quality assurance testing of contact center projects for multiple clients.
"The kind of growth we have experienced this year, our 30th year in business, warranted an investment in new offices," said Barry Hays, senior vice president of TeleVoice. "This expanded space enables us to add technology and marketing staff to our team, These additions will help us continue to provide the highest level of service as our client base continues to expand.
Posted by Veronica Silva Cusi, news correspondent
Source: http://www.businesswire.com
Related Editorial
More Editorial From TeleVoice
About TeleVoice:
Established in 1986, TeleVoice delivers customized telephony solutions, including Interactive Voice Response (IVR) and Computer Telephony Integration (CTI) to the financial services industry. Based in Houston, Texas, TeleVoice has been the call center solutions partner of industry leader Black Knight Financial Services, formerly LPS, since 1987.
Published: Thursday, September 1, 2016


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Live on SS2 & BT2: Sat 12.15 & 17.30: An Inverness win over Aberdeen and a 10/1 (bet365) draw double are among our Scottish picks.
It's top against bottom as Celtic entertain Hearts in the early kick-off. Few would bet against the hosts taking maximum points here but a victory for the champions won't pay off the mortgage as Neil Lennon's boys are just 1/8 at best with Paddy Power to heap more misery on the beleaguered Jambos, bet365's 20/1 underdogs.
The Hoops hammered Hearts 7-0 in the cup recently and they're 80/1 at BetVictor for a repeat but punters can expect the Gorgie outfit to be a touch more resilient on this occasion so a repeat of the 3-1 Bhoys triumph from earlier in the campaign looks more realistic at 12/1 via Coral.
With Inverness and Aberdeen both right in the hunt for second spot their derby showdown takes on extra significance and as Caley Thistle have won five of their last six in the league, it's they who are expected to emerge victorious, BetVictor installing them as 31/20 favourites.
The Dons, who shaded their Pittodrie contest 1-0 earlier this season, will have their backers at 2/1 (Betfred) but home advantage could prove crucial and the hosts stand out at 13/4 via Ladbrokes to prevail by one.
In-form Dundee United can't stop scoring and it's hardly surprising that they're just 2/5 at best with Stan James to send off-colour Ross County home from Tannadice empty-handed. Possessing all the hallmarks of a home banker, the Terrors have racked up an impressive 21 goals in five league and cup assignments and they're 7/5 at Unibet to net at least three times in this one.
Fifth-placed Motherwell will consider their home game against St Mirren as a chance to gatecrash the battle for Europe and they're 10/11 at Winner.com to do the business at Fir Park.
St Johnstone and Hibs sit either side of the divide in the middle of the Premiership table, although Saints will move seven points clear of Terry Butcher's troops if they triumph in the pair's clash at Easter Road. The visitors are 6/4 at William Hill but home advantage should ensure the Hibees nick a point (5/2, bet365).
Meanwhile, neither Kilmarnock or Partick can lay claim to any kind of form at present and it's impossible to pick a winner at Rugby Park, where they're 12/5 with Coral to be inseparable.
Recommended Bets:
Inverness to beat Aberdeen by one – 13/4 (Ladbrokes)
Double on draws at Hibs and Kilmarnock – 10/1 (bet365)
Celtic to beat Hearts 3-1 – 12/1 (Coral)
Dundee Utd to score over 2.5 goals – 7/5 (Unibet)
Compare all Scottish Premiership odds and bet at the best price.


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This star rating is the opinion of the property owner only. A Self-Rated hotel has not been assessed by the AA or Visit Britain, whose official star ratings we use across LateRooms.com.
Best Western Hotel Pythagorion3 Self rated
28, Agiou Konstandinou street, Athens, 10437
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About the hotel
PYTHAGORION HOTEL, Athens, Greece is located next to the National Theater and conveniently situated within walking distance of Plaka area the old quarter of Athens, Acropolis, the Parthenon, Syntagma Square, the Main Railway station and the Metro of Athens. Hotel Pythagorion provides an ideal base for the Athens visitor with easy access to most of Athens sights and attractions as well as the dining and shopping areas. The hotel offers High Speed FREE INTERNET CONNECTION from your own room using your own laptop and High Speed FREE INTERNET CORNER with FREE USE OF PRINTER!!!!!!! Free welcome drink to all guests.
Free Orange juice throughout the day to all guests MAPS of Athens and BOOK with general information's about Athens, FREE from our INFO Stand . 24-hour reception, and the friendly staff is always ready to give you advice on what to see and do in Athens. Multilingual Staff Credit Cards accepted: Visa, Mastercard, American Express,Diners Wake up calls Doctor Assistance Luggage storage Elevator Car rental desk Transportation facilities upon request Information for sightseeing tours and island cruises Meeting facilities Fax services Photocopy services Children under 2 years old - Free of charge Iron and Ironing board available Daily House keeping . Hotel Rooms: 56. Hotel Chain: Best Western.
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text:
Environmental services and equipment provider, MSIS has strengthened its board with the appointment of a new non-executive chairman.
John Forrest, whose career in the upstream oil and gas industry spans more than 35 years, joins the MSIS Board later this month (July 2015).
Currently chairman of PDG Helicopters and Task Fronterra, Mr Forrest has provided Board-level leadership to several fast growing companies and has broad experience of the upstream section, in particular in HSE leadership, asset management and operations. He is a past chairman of the Industry flagship organisation Step Change in Safety.
Chris Lloyd, MSIS Chief Executive, said: "We are extremely pleased to welcome John onto the MSIS board. His background and hands-on experience in the industry, together with his diverse management experience, provides an ideal profile for the post.
MSIS Group
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Ian Stuart /
Ian Stuart 'Rossini'
/ Resell It & Make Up To: $2,400 Learn More
This is a "plus size" couture wedding gown. Ian Stuart does not make dresses this size regularly. The color is taupe with ivory underskirt and multicolored floral accent at side bustle. Worn for only 4 hours and the measurements are bust 44 inches,waist 40 inches, and hip is 40 inches. Cleaned right after wedding. Cathedral length train with french bustle, very easy to bustle and there are straps with corresponding numbers. Many buttons up the back.
Once you have placed your order the Seller will be notified to pack and ship the gown. The seller will then ship the item to us and our inspection and authentication team will check that it is authentic and matches to the seller's description. The item will then be shipped to you upon approval.
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Try It On - No Risk Return Policy: Have your cake & eat it too. Returns accepted on all items. More details


text: Cash Warren took Instagram to share a sweet shot while vacationing in Hawaii with his wife Jessica Alba and their family. In the shot, his children Honor and Haven look happy while they both enjoy a bowl of ice-cream. Alba shared a different shot of the kids on … [Read more...]


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- English
- français
Emergent Connectivity Principles in the Neocortex
The neocortex makes up over 80% of the mammalian brain and is responsible for higher cognitive functions, processing of sensory inputs and orchestration of complex motor outputs. It is a 6-layered structure composed of billions of morphologically and electrically diverse neurons. Functionally, the basic unit is the neocortical column (NCC), a vertical structure of 0.5 mm wide, repeated millions of times across the neocortex and connected in an intricate but consistent way. In this thesis I investigated the role of morphologies (neurogeometry) in shaping the elaborate connectivity scheme within a column. First, I suggest a morphological basis for the higher than expected reciprocal connectivity reported experimentally between connected pairs, using a newly defined measure: the Reciprocity Index (RI), which arises from a purely mathematical concept and can be derived from the morphometric statistics of neurons. Second, I show that most experimentally reported synaptic patterns between different classes of neurons can be directly computed from the statistics of their morphologies, while some pathways would require additional functional mechanisms to refine an even more specific synaptic pattern. My thesis is done within the Blue Brain Project, the first comprehensive attempt to reverse-engineer the mammalian brain, starting with the somatosensory NCC of a P14 rat. I present in this thesis my contribution in constructing the framework for building biologically accurate circuitry. I explain how we start from 3D neuron morphologies obtained in vitro, repair them for slicing artefacts, build circuits and accurately detect potential synapses. I also explore how modelling of the experimental procedures can help us characterize biases and predict in vivo data from in vitro data. I finally present recent exploratory work on how to use the supercomputing power to design novel in silico protocols to investigate the emergent dynamics of the neocortical column.
Keywords: neocortical column ; connectivity ; neurogeometry ; morphologies ; emergent properties ; primary somatosensory cortex ; large scale modelling ; simulation driven research ; synaptic pathways ; reciprocal connection ; reciprocity index ; connection probability profile ; in silico ; colonne néocorticale ; connectivité ; neurogeometry ; morphologie ; propriétés émergentes ; cortex somatosensoriel primaire ; modélisation à grande échelle ; recherche axée sur la simulation ; connexion réciproque ; indice de réciprocité ; in silico ; voies synaptiquesThèse École polytechnique fédérale de Lausanne EPFL, n° 4631 (2010)
Programme doctoral Neurosciences
Faculté des sciences de la vie
Institut des neurosciences
Laboratoire de neuroscience des microcircuits
Record created on 2010-01-18, modified on 2017-05-12


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Everyone needs a method of storing leftovers. That may be why, slowly but surely, a collection of takeout containers has amassed in your kitchen. The only trouble is that different containers from different sources rarely match in size. That makes it difficult to use the limited space in your fridge most efficiently by stacking containers on top of one another. The simple secret: Buy sets of containers that are all the same size. The plastic ones come cheap, but you can actually save money if you invest in containers made of glass. Unlike plastic, glass doesn't stain or absorb odors. Handle yours carefully, and they can last forever. Available at Amazon; $10.99 per set of three 12-ounce containers.
Related: 10 Instant Cures for Any Kitchen Odor
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my power outfit
rachel ienna
My power outfit revolves around my NY tank and old Pistons hat. I love to pair it with my plaid flannel, leggings, and either Converse or Uggs depending on the weather. The outfit is inspired by a mix of 90s grunge and current trends. I would consider this my power outfit because it's comfy and casual, but it also flatters my figure and gives me confidence.


text:
Question: How does internal auditing differ from external auditing
How does internal auditing differ from external auditing?
View Solution:
View Solution:
Answer to relevant QuestionsMultiple Choice Questions:1. To what does the word forensic pertain?a. Financial audits when fraud is involved.b. A particular type of accountingc. Matters relating to law and courts of law.d. Prosecutorial investigations.2. ...Westag Mills Company is considering a merger with a competitor. Westag's CEO has hired a forensic accounting firm to investigate the competitor. What types of investigations could the forensic accounting firm conduct? What ...1. In a criminal investigation, the prosecutor represents the interest of the defrauded party.2. In a criminal case, there is no constitutional right to a jury trial.3. A state constitution may not override the U.S. ...Why is bail required? Are there any mitigating factors that could decrease the amount of bail? Are there any charges for which there is no bail? What protection does a bail bond seller have?Multiple Choice Questions:1. Approval applies to transactions thata. Have been not yet been authorized.b. Are pending authorization.c. Are fully authorized.d. All of the above.2. Segregation of duties applies to separating ...
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15 Reviews
Grab a ride by the tail.
On long, hot days in the saddle, our hands are often the first casualty, but Giro's Tessa Women's Gloves incorporate mesh backs and a healthy hit of padding in the synthetic palms to shield against sore grabbers without the risk of overheating. The palms themselves are durable enough to stay on the hoods for as long as we're able to stay in the saddle, and they provide a bit more grip than skin alone.
- Lightly padded synthetic Clarino palm
- Mesh backs
- Hook-and-loop strap closure
- Item #GIR0356
Tech Specs
Gearhead Help
Tech Specs
- Reviews
What do you think about this product?
Have questions about this product?
Fits great!
- Familiarity: I've used it several times
- Fit: True to size
- Size Bought: Small
- Height: 5'2"
- Weight: 112lbs
Nice glove, I use for long road rides to protect my hands, they make my rides more comfortable. They have a great fit.
Nice Gloves
- Familiarity: I've used it once or twice and have initial impressions
- Fit: Runs small
- Size Bought: Medium
- Height: 5'4"
- Weight: 130lbs
These gloves definitely make riding more comfortable and fit decent, but are a little tight. They're a bit hard to get off once you're done riding and sweaty, but other than that they're a decent glove. I'm hoping they may stretch out once I used them a bit more, but overall they improve my comfort while riding.
Run small
Nice gloves but run small. I'm re-ordering in a larger size.
Finally a pair that fits!
- Familiarity: I've used it once or twice and have initial impressions
- Fit: True to size
- Size Bought: Large
I have only used these gloves a handful of times so I cannot speak to the longevity, but they have greatly improved my overall comfort while riding. There is enough gel to dampen the vibrations of the road, but not enough to change my natural hand position on my handlebars. I also have noticed how breathable the fabric is. My biggest issue with finding gloves was finding a pair that actually fit my hands. I was hesitant to purchase men's, as I have small wrists , so I was ecstatic to find a pair of women's that not only fit but performed exceptionally well. As long as you take the time to look at the size chart above, you should be able to easily determine the ideal size you need. I have read reviews where some customers have experienced the fabric tearing at the seams. I do not know if they are defects, but I have stuck with hand washing them and have not had any issues.
pretty comfy
- Familiarity: I've used it several times
- Fit: True to size
This are pretty comfy..I ordered a medium and I think they fit well. I wish they had the pull loop on the fingers to get the gloves off after a long sweaty ride and they are already showing dirt stains. I got the grey white so i feel that's expected with such a light color. Other wise my hands feel great on the bike!
- Familiarity: I've put it through the wringer
- Fit: True to size
I own multiple pairs of these gloves. The padding is a little too stiff on the first ride, but after a ride or two of break-in, they're perfect.
Run small
- Familiarity: I've used it once or twice and have initial impressions
- Fit: Runs small
- Size Bought: S
- Height: 5'2"
- Weight: 128lbs
I purchased these for someone who wears smalls but she said these were a little tight. She kept the gloves but I don't know how much wear they'll get. Although, they should loosen up after being worn a few times. We'll see.
Bought a second pair
- Familiarity: I've put it through the wringer
- Fit: True to size
- Size Bought: Small
- Weight: 110lbs
My dog chewed up a glove and I bought a second pair, wouldn't wear anything else.
COOL gloves
- Familiarity: I gave it as a gift but have feedback to share
- Fit: Runs small
- Size Bought: S
- Height: 65'65"
- Weight: 143lbs
Nice glove is a little next time you buy M code,I am customers from China
Customer Loves them!
Feel great!!
- Familiarity: I've used it several times
- Fit: True to size
I'm relatively new to riding, but heard that riding with gloves would make a huge difference in my hands. After riding with these gloves, I can't imagine ever riding without them again! They're amazing! Extremely comfortable...the padding is perfect! They fit great, are easy to get on and off, and washed up perfectly! I definitely recommend!
Couldn't But Enough Pairs!
- Familiarity: I've put it through the wringer
- Fit: Runs small
Purchased several of these for family and friends because of how much my wife liked them. They have an excellent fit and grip.
Giro tessa gloves. white. large
- Familiarity: I've put it through the wringer
- Fit: True to size
- Size Bought: large
- Height: 5'9"
- Weight: 165lbs
I'm 5'9" and the large fits. I measured my hand like the sizing chart suggests and they're true to size. I ride a lot and so yes, they got stinky after a few weeks, but washed great.
I loved the close-out pricing! Thanks!
Like 'em
- Familiarity: I've put it through the wringer
- Fit: True to size
These gloves get the job done. They are breathable, look good, have adequate padding, wash easily, and don't get stinky.
The only con: They are not as grippy as I'd like. Sometimes on a descent when I'm braking and hit a bump, my hand flies off of the handlebar and brake. Part of this could be due to the exposed fingertips versus full fingered gloves, but I'd like for the palm of my hand to stick to my bike grips too.
Tessa Glove in Action
Nice Glove
- Familiarity: I've used it several times
Comfortable riding glove...washes well and comes out soft and flexible. Good deal on a good glove.
My all-time favorite gloves
I LOVE these gloves. I have no complaints about them, whatsoever.
They provide adequate padding for longer rides. They are comfortable and well-constructed. Gloves are true to size. I like mine to be a little roomy, so my hands have space to swell on hot days when my hands get sweaty. The thumb has a great little suede-like piece that is great for wiping sweaty sunglasses.
I purchased mine nearly a year ago and wear them on almost every ride, and the padding just started to wear out recently. I ordered 3 more to have for future use, just in case Giro discontinues these.
I have bought over half a dozen gloves in the past couple of years, and these are, hands down, the winners.


text:
Mahesh and Charan are having this habit of clashing at box office on the eve of Sankranthi. On two occasions earlier, one during 2013 they locked horns with Nayak and SVSC, and then in 2014 they confronted each other with 1-Nenokkadine and Yevadu. Nayak worked slightly better than SVSC at box office giving some edge to Charan in 2013. And on their next clash too, the young hero emerged triumphant with 1-Nenokkadine turning disastrous and Yevadu reaching 50 Cr mark. But for 2014, the race is not yet over as Mahesh and Charan are competing again.
Reports have that Sreenu Vytla directed 'Aagadu' is being planned for September 26th release, while Krishna Vamsi's latest 'Govindudu Andarivadele' will also hit screens on same day. Mahesh will be seen as encounter specialist police officer in this Dookudu Sequel whereas Charan is coming up with a complete sentimental family story.


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Nan Du
My research focuses on developing large-scale machine learning models and algorithms for statistical analysis of temporal/spatial dynamics arising from networked systems, which has practical applications in promoting online user engagement, optimizing advertisement allocations and providing context-aware recommendations.
Google Publications
Coevolutionary Latent Feature Processes for Continuous-Time User-Item Interactions
Yichen Wang, Nan Du, Rakshit Trivedi, Le Song
Neural Information Processing Systems (2016)
Dirichlet-Hawkes Processes with Applications to Clustering Continuous-Time Document Streams
Nan Du, Mehrdad Farajtabar, Amr Ahmed, Alexander J. Smola, Le Song
Proceedings of the 21th ACM SIGKDD International Conference on Knowledge Discovery and Data Mining (2015), pp. 219-228


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FLAGSTAFF, Ariz. -- There's a subtle grin and confidence that emerges when Ken Whisenhunt acknowledges all the questions about the Arizona Cardinals' quest to win their third consecutive NFC West title while many others believe it's the year of the San Francisco 49ers.
"Our team has played pretty good football when people write us off," the Cardinals' coach said. "If we respond the way we have in the past, I think we'll be pretty happy."
Then again, those Cardinals teams responded with Kurt Warner playing quarterback, Anquan Boldin playing wide receiver, Karlos Dansby playing linebacker and Antrel Rolle playing safety. They're all gone in 2010.
Yet Whisenhunt is quick to draw from his experience as an assistant coach with the Pittsburgh Steelers to never push the panic button.
"With the Steelers, you lost a Rod Woodson at safety and a Carnell Lake stepped up . You'd lose a Joey Porter but you find a James Harrison," Whisenhunt said.
"If you're a good football team, you go through those transitions and keep going. Look, when Anquan Boldin was hurt, Steve Breaston stepped in and we kept winning. Now it's time for some young guys like [defensive end] Calais Campbell, [cornerback] Greg Toler, [wide receiver] Early Doucet, [running backs] Beanie Wells and Tim Hightower to step their games up."
It's also no coincidence that Whisenhunt did not shy from bringing in two former Steelers greats, guard Alan Faneca and outside linebacker Porter, despite the perception that their better days are behind them.
"Oh, there's a lot that Joey and Alan bring -- they've been two of the better players in the league, they've played on championship teams and they've played in Pro Bowls," Whisenhunt said. "When you go through a transition with young players, they can look at [Faneca and Porter] and see that this is how you work, this is how you prepare. They're going to have a very positive impact on our young players and our team."
The glaring question mark is at quarterback. Matt Leinart is the starter, but even Whisenhunt pointed out that Derek Anderson has shown the talent few quarterbacks in the NFL possess, especially with the offensive skill talent around him headed by All-Pro wideout Larry Fitzgerald.
"Matt learned a lot from Kurt -- I'm excited about him and I'm excited by Derek Anderson, who has been to a Pro Bowl and understands the game," Whisenhunt said. "I think everyone knows we don't shy away from competition. Go back three years ago, and I don't think anyone thought Kurt was going to be the guy but he won the battle with Matt.
"That's what our preseason is all about. What kind of team are we going to be? Will we run the ball more with Beanie and Hightower? Can we still throw the ball special with the receivers we have? I think we're all waiting to see what this team is all about and who's going to step up."
Whisenhunt did offer a vote of confidence for Leinart, even if there is a reality that Anderson is capable of overtaking the job at some stage.
"I think Matt's really matured," the coach said. "I think what he's been through has made him stronger."
Leinart himself conceded Anderson is a worthwhile competitor but won't panic on his bad days.
"I know DA can really throw it and has a big arm, but I just spent three years battling a Hall of Fame quarterback," Leinart said.
"I know this really is my job for the first time but I'm not assuming anything. People can say what they want but I'm also not looking over my shoulder no matter what. One of the things I learned from Kurt is that he had days and games where he struggled but he'd bounce back. Trust me, I learned a lot [from Warner] and I'm here every day to just do my job and lead this team that has a lot of talent around me."
Other observations from Cardinals camp:
Whisenhunt conceded he's "had a number of discussions" with Warner after his January retirement but never really believed the quarterback would pull a Brett Favre and come back for another season.
While Leinart will have to play well to keep Anderson on the sidelines, the No. 3 quarterback battle is another camp story because undrafted free agent Max Hall (BYU) has been very impressive and could beat out fifth-rounder Fordham QB John Skelton, who shows a strong arm. Why is it noteworthy? The Cardinals believe Hall has some Warner-like abilities despite being undersized.
Hightower and Wells both seem strong at running back; Wells looks a little lighter, and his big-play ability is very evident.
Of all the defensive players teammates and coaches say could have a Pro Bowl, breakout year is 6-foot-8, 300-pound Campbell, a third-year defensive end who emerged as a force last season.
Fitzgerald's Twitter following is so large (more than 500,000) that VP media relations director Mark Dalton utilized the receiver's outreach on the social network when the team began its season-ticket push.
ESPN senior NFL analyst Chris Mortensen will file reports from all the training camps and send updates on the road via Twitter (@mortreport).


text:
Download SCCL Mines Admit Card/Hall Ticket for Management Trainee 2015 Print Admit Card Check Exam Date Singareni Collieries Company Limited Executive and Non Executive Posts Exam SCCL MT (E&M) Admit Card 2015 Check admit Card Exam Date
SCCL Exam Admit Card 2015
LATEST UPDATE: Admit Cards for SCCL Management Trainee (E&M) are available for Download Now. Candidates can Download Admit card from the link provided below………….
Singareni Collieries Company Limited is going to conduct written test for the recruitment to 1178 Executive and Non-Executive Cadre posts. Exam for these posts will be held in upcoming dates.
DATE OF SCCL MT (E&M) Exam : Sunday, 28 June, 2015
Download Model / Sample Question bank for Assistant Foreman Trainee (Electrical)
Download Model / Sample Question bank for Management Trainee (CIVIL)
Download Model / Sample Question bank for Management Trainee (Mining)
Admit Card : Candidates who applied for this Competitive Exam should download their E-admit Card or Hall Ticket to enter in prescribed Exam Centre of these various posts Exam conducting by SCCL. No Admit Card or Hall Ticket will reach home of Candidates by post, it can only be download in upcoming dates before exam for following posts :
Executive Trainee Cadre :
→ Management Trainee (Mining)
→ Management Trainee (E&M) (Mining)
→ Management Trainee (Civil)
→ Management Trainee (Personnel
→ Management Trainee (F&A)
Non Executive Cadre :
→ Junior Mining Engineer Trainee (JMET)
→ Assistant Foreman Trainee (Mechanical)
→ Assistant Foreman Trainee (Electrical)
NOTE : The minimum qualifying marks in case of Executive cadre posts is 40% for unreserved candidates, 35% for BC candidates and 25% for SC/ST candidates and for Non – Executive Cadre 35% for unreserved candidates, 25% for BC candidates and 15% for SC/ST candidates.
SCCL Management Trainee E&M Admit card 2015
Click Here to DOWNLOAD MANAGEMENT TRAINEE (E&M) ADMIT CARD 2015 (Available Now)
Click Here to DOWNLOAD Assistant Foreman Trainee (Electrical) ADMIT CARD 2015 (Available Now)
Click Here to DOWNLOAD MANAGEMENT TRAINEE (CIVIL) ADMIT CARD 2015 (Available Now)
Click Here to DOWNLOAD MANAGEMENT TRAINEE (MINING) ADMIT CARD 2015 (Available Now)
Click Here to DOWNLOAD JUNIOR MINING ENGINEER TRAINEE (JMET) Hall Ticket 2015 (Available Now)
Caution : Mobile phone and other electronic gadgets of communication are strictly banned in the campus of the examination hall.
Candidates should have to keep in touch with Website of Singareni Collieries Company Limited (SCCL) for more updates regarding information about Exam and its Admit Card.
For more Information please visit – http://scclmines.com/scclnew/careers.asp
For Any Query or Doubts Candidates can Leave Comment in the Comment Box. We will Try to give Appropriate Response of Queries and Clarify Your Doubts.


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These days, kurtis have turned out to be extremely mainstream in the midst of young ladies and ladies of all age bunches. Kurtis give a slick and enchanting look to your persona furthermore give you awesome level of solace Be it in school, at business sector, at office or just meandering out with companions or family, kurtis give you a great deal of simplicity and comfort. In modern fast life, kurtis have become all time hit apparel for many women. Not everyone is blessed with enviable height like that of B'wood's Deepika and Bipasha. But hey, being short is not so bad either. If you are big on personality and style, no one will even bother about your lack of height. After all, good things come in small packages! Many a times we see how short girls find it really difficult to style long kurtis as they have a mindset that they would look even shorter in that case, but trust me even you can make it look stupendous by styling them in differently. So here we have some styling tips on long kurtis for short height girls.
1. Pick Elongated Patterns On The Kurta-Long Kurtis For Short Height:
Ensure you pick prints that are not making you look massive. Paisley prints, stripes, long themes are what you ought to search for when you purchase kurtas. Kurtas that have vertical stripes or vertical plackets along the inside will consequently make you look taller.But horizontal strips are totally a big NO for you.
2. Wear Long Flowy Kurtas With Palazzos-Long Kurtis For Short Height:
While wearing A-line long kurtas and anarkalis, pair them with flowy palazzos. They are absolutely in vogue and are complimenting for each body shape. It takes away consideration from the stature and makes your outfit look overwhelming.
3. Remember To Wear Tall Heels With Churidaars/ Slim Pants-Long Kurtis For Short Height:
In the event that you are wanting to wear churidaars or slim pants with a long kurta, bear in mind to wear them with stilettos that have thin straps or wedges if you are looking for comfort. Indeed, even a tiny bit of heel can have a gigantic effect to your look. This consequently makes one look taller, and with a straight fit kurta, the visual appeal is much higher.
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By now as we have read so much on this topic, i'am definitely sure that you very well know by now that straight kurtas are totally your thing. . Straight cuts, linear or tiny print patterns make one look lean. Thus when choosing long kurtas, keep this in mind and just say a big NO to horizontal lines and big patterns.
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Skip to comments.American Psychological Association Ignores Abortion-Depression Link
Posted on 08/14/2008 8:32:39 AM PDT by julieee
Washington, DC -- A panel of the American Psychological Association released a new report Wednesday that came to the conclusion skeptics predicted. The organization, which supports legalized abortion, claims abortion causes no mental health problems despite recent research proving otherwise.
The group's conclusions aren't a surprise given that pro-life advocates and a psychology professor both noted the organization stacked the panel with abortion advocates.
(Excerpt) Read more at LifeNews.com ...
Of course they do. They're a political group, after all.
Exactly - they voted to have Homosexuality removed as a disorder, solely as a politically correct (or actively activist) expedient.
Wish there was some way to get around this; hold them accountable or form a new, effective, organization.
They are wrong.
According to the liberal fascists and other pro-choice Romans, one should feel no remorse for extinguishing innocent life and using the body parts for the common good.
And imagine how this organization will morph under an Obambi administration! Food for thought...
Similarly, the American Pyschological Association went on record against life and on the the side of depression and death in the 1970s, as a result of a few homosexuality activitists who gained power within the organization.
Analogous to this pro-abortion stand, the Association's offical voice was maneuvered by those with a political agenda to declare that homosexuality was no longer something to be treated and is perfectly normative. With that vote, homosexual attraction and behavior were summarily taken off the official list of mental illnesses.
Instead mental health practitioners were told that even when people are seeking reparative therapy for unwanted same sex attractions, the patients should be convinced they should stick with and be happy with their homosexual attractions, this despite evidence that homosexual behavior leads to more depression and suicide than the choice to work toward heterosexuality.
Political agendas rather than science are still driving the APA.
Recover it's 'health,' or let it slide into irrelevancy and replace it with a properly functional equivalent?
Why the drop after 1960? (in deaths of women from illegal abortions)
The reasons were new and better antibiotics, better surgery and the establishment of intensive care units in hospitals. This was in the face of a rising population. Between 1967 and 1970 sixteen states legalized abortion. In most it was limited, only for rape, incest and severe fetal handicap (life of mother was legal in all states). There were two big exceptions California in 1967, and New York in 1970 allowed abortion on demand. Now look at the chart carefully.
Abortion Statistics - Decision to Have an Abortion (U.S.)
· 25.5% of women deciding to have an abortion want to postpone childbearing
· 21.3% of women cannot afford a baby
· 14.1% of women have a relationship issue or their partner does not want a child
· 12.2% of women are too young (their parents or others object to the pregnancy)
· 10.8% of women feel a child will disrupt their education or career
· 7.9% of women want no (more) children
· 3.3% of women have an abortion due to a risk to fetal health
2.8% of women have an abortion due to a risk to maternal health
So how many womens lives have been saved by abortion?
Only about 3% of abortions since 1972 were reported to be due to a risk to maternal health. A reasonable person would recognize that not all of those cases represent a lethal risk. But lets say they did. That means that nearly 45 million fetuses were butchered to save the lives of about 1.3 million women. Or put another way; 35 babies are killed to save each woman.
Abortion was legal in all 50 states prior to Roe v. Wade in cases of danger to the life of the woman.
Roe v Wade: FULL Text (The Decision that wiped out an entire Generation 33 years ago today)
Disclaimer: Opinions posted on Free Republic are those of the individual posters and do not necessarily represent the opinion of Free Republic or its management. All materials posted herein are protected by copyright law and the exemption for fair use of copyrighted works.


text:
From the Center for AIDS Prevention Studies (Drs Lurie, Chesney, and Lo), Department of Medicine, AIDS Program (Dr Hecht), Center for AIDS Research (Dr Chesney), and Division of General Internal Medicine, Program in Medical Ethics (Dr Lo),University of California, San Francisco; Public Citizen's Health Research Group, Washington, DC (Dr Lurie); and the Maternal and Child Health Program, School of Public Health, University of California, Berkeley (Dr Miller).
In the wake of recent breakthroughs in antiviral therapies and Centers
for Disease Control and Prevention (CDC) recommendations advocating occupational
postexposure prophylaxis (PEP), health care workers are increasingly receiving
inquiries about PEP following exposures to the human immunodeficiency virus
(HIV) through sex and injection drug use. The probability of HIV transmission
by certain sexual or injection drug exposures is of the same order of magnitude
as percutaneous occupational exposures for which the CDC recommends PEP. In
such cases, if the exposure is sporadic, it seems appropriate to extrapolate
from the data on occupational PEP and recommend prophylaxis. However, for
individuals with continuing or low-risk exposures, we instead recommend referrals
to state-of-the-art risk reduction programs. Clinicians, using local HIV seroprevalence
data and their knowledge of transmission probabilities, can help exposed patients
make an informed decision regarding PEP. Because of the large number of risky
encounters that will not be treated prophylactically, even after significant
outreach efforts, public health interventions that emphasize PEP as part of
a comprehensive HIV prevention program should be confined to cities with highest
HIV prevalences.
Lurie P, Miller S, Hecht F, Chesney M, Lo B. Postexposure Prophylaxis After Nonoccupational HIV ExposureClinical, Ethical, and Policy Considerations. JAMA. 1998;280(20):1769–1773. doi:10.1001/jama.280.20.1769
© 2017


text:
Tagalog for Beginners: Learn how to describe Yourself plus your Family and Friends
Jonnalyn Camba, Entrepreneur, Language & Technology Instructor
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Personal Pronouns3:31
Combining Adjectives & Personal Pronouns2:29
About This Class
In this lesson, we will go over personal pronouns and adjectives so that you will be able to combine them to create sentences.
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Hi. I'm Jonnalyn, a persistent learner just like you. Before I started teaching online classes, I had been taking online classes for many years. I realized the value of online learning right away and wanted to become part of this revolution where anyone can learn whatever they desire.
Here are the other classes I'm teaching: https://www.skillshare.com/jonnalyncamba
Some fun facts about me are: I have a dual major in Economics and Psychology, I love to travel and eat diverse dishes, I love learning languages, I love to write (especially poetry), and I hula hoop really well.
My principle in teaching is to provide my students a fun and simple way to learn anything. I believe the journey is just as important as the destination, which is why I do my best to create beautiful courses that my students can refer back to again and again.
The details and nuances in learning are important, but keeping the big picture in mind for guidance is equally necessary. I am mindful of this in whatever I teach.
My ideal student is the persistent learner who loves learning for learning's sake and sees the world not only in practical terms, but in its poetic, mystical beauty.
Join me in my Tagalog classes, technology, and other artsy classes that are designed for you to enjoy again and again.
Best wishes!


text: RT Book, Whole DB /z-wcorg/ DS http://worldcat.org ID 46640561 LA English T1 Downtown : its rise and fall, 1880-1950 A1 Fogelson, Robert M., PB Yale University Press PP New Haven YR 2001 SN 0300090625 9780300090628 0300098278 9780300098273 AB "Written by one of this country's foremost urban historians, Downtown is the first history of what was once viewed as the heart of the American city. It tells the fascinating story of how downtown - and the way Americans thought about downtown - changed over time. By showing how businessmen and property owners worked to promote the well-being of downtown, even at the expense of other parts of the city, it also gives a riveting account of spatial politics in urban America. Drawing on a wide array of contemporary sources, Robert M. Fogelson brings downtown to life, first as the business district, then as the central business district, and finally as just another business district. His book vividly recreates the long-forgotten battles over subways and skyscrapers in the late nineteenth and early twentieth centuries. And it provides a fresh, often startling perspective on elevated highways, parking bans, urban redevelopment, and other controversial issues. This groundbreaking book will be a revelation to scholars, city planners, policymakers, and general readers interested in American cities and American history."--Jacket.


text:
If I report different dependents than I claimed in the prior tax year, or have added additional dependents to those that I claimed last year, should I expect to be audited?
The IRS doesn't disclose the standards and criteria used for its dependent eligibility audit selection.
You can claim any person as a dependent if he or she meets the requirements for a qualifying child or a qualifying relative.
There's no maximum number of dependents you can claim.
Ready to file?
You've got your forms. And we've got your back. There's only one thing left to do. Let's do this.
So how much will you get (or owe) this year? That's the million-dollar question. We happen to have three very useful calculators to help you estimate your refund or balance due.


text:
Each month, Rated People connects over 50,000 homeowners with quality tradesmen - simply enter your job details in the form above to get started.
We'll match your job to the relevant tradesmen in your area - those interested will pay to see your contact details and call you to quote.
As each tradesman expresses interest in quoting on your job, we'll send you their profile and ratings to help you choose the right one for you.
After the work is complete, return to the site and rate your tradesman based on their quality of work, reliability and value for money.
Added 11/09/2015
I require a new radiator in my bathroom, the old one is starting to rust. It could be tied...
Added 04/10/2014
*PRIORITY LEAD* Changed information:description, budget Services required: Flat roof replacement...
Added 16/04/2012
My Mira shower has stopped working and I want to get it repaired.
Managing your job is easier with the Rated People Homes App, download it today.


text:
The LEGO Book - Hardcover Book
Discover all there is to know about one of the world's most beloved toy companies.
Beautiful pages reveal, explore and celebrate the fascinating story of LEGO® bricks and the history behind them in this newly updated edition of The LEGO Book. Along with 56 new pages, a timeline highlights key moments in LEGO history and special features spotlight numerous groundbreaking and momentous achievements. Includes new and updated information about LEGO Star Wars, LEGO Ninjago, LEGO Friends, and much more.
Explore all the main LEGO toy systems, the LEGOLAND® theme parks with their amazing animated models, and the LEGO Group's spectacular diversification into films, video games and the visual arts. Incorporating clear instruction panels on how to create great models from just a few simple bricks, The LEGO Book is truly a treasure perfect for any LEGO fan!
256 pages
Recommended for ages 8 years and up
Dimensions: 10.9 x 9.3 x 1 inches
*member price, for regularly-priced merchandise only:
Please enter your membership number at checkout. To become a member, please visit our Membership site.
Proceeds from CAF's tours, programs and retail shop support the educational mission to inspire people to discover why design matters.
We Also Recommend


text: DescriptionEdit A full-metal mace with a short shaft and attached blades. The smith who forged this weapon certainly knew what he was doing. Source / Where can it be foundEdit Settlement weaponsmith. Loot in battle.(needs confirmation) NotesEdit Retrieved from "http://battlebrothers.wikia.com/wiki/The_Club?oldid=6029" Ad blocker interference detected! Wikia is a free-to-use site that makes money from advertising. We have a modified experience for viewers using ad blockers Wikia is not accessible if you've made further modifications. Remove the custom ad blocker rule(s) and the page will load as expected. Categories: Melee Weapon Add category Cancel Save


text:
Need a walk in lab test in Alexander City, AL? Simply order affordable testing online and get a walk in lab test in Alexander City, AL (Find More Locations):
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text:
Definition Edit
"Organizations, such as DoD organizations, may choose to develop internal standards documents that are specific to the organization (DoD Internal Standards). The BMO generally does not recommend use of internal standards for DoD biometric deployments, except in cases in which the recommended types of standards . . . do not address the standards issues in question. Use of ISO standards, ANSI standards, and NIST standards are the preferred methods of applying standards to DoD biometric programs."[1]


text:
Toyota of Puyallup
Review for Toyota of Puyallup
KELLY P. on 08/12/2015
"Customer Service"
Great customer service in Parts & Service Departments, as well as on the phone.
Vehicle: TOYOTA MATRIX
Category: Service
Service Date: 08/10/2015
Review Created: 08/12/2015 04:27 PM
Kerry S., President & General Manager at Toyota of Puyallup responded on 08/12/2015
Thank you for your valuable feedback, Kelly. We appreciate your business, and this is great to hear. See you next time!
President & General Manager
Toyota of Puyallup


text:
What is JotForm?
JotForm is a free online form builder which helps you create online forms without writing a single line of code. No sign-up required.
At JotForm, we want to make sure that you're getting the online form builder help that you need. Our friendly customer support team is available 24/7.
We believe that if one user has a question, there could be more users who may have the same question. This is why many of our support forum threads are public and available to be searched and viewed. If you'd like help immediately, feel free to search for a similar question, or submit your question or concern.
Downloading/Accessing photos uploaded in my form is blocked by Comodo Secure DNSAsked by organizasyonservisi on March 05, 2015 at 03:24 AM
hello!I can not see the photos uploaded my form. I get an error message. I would like to help in this matter. thanks
JotForm Support
I used Comodo Secure DNS, but accessing and downloading files from a jotform are working for me. I used the same DNS for over a year now. What browser is that? Let me know and we'll check further. Are you getting the same error for the other browsers? Please try it.
chrome 41.0.2272.76
did not improve
JotForm Support
Are you using Comodo Internet Security Suit? If so, I would suggest adding Amazon AWS to your whitelist settings.
Check this guide: https://help.comodo.com/topic-72-1-522-6408-.html.
Thank you!
I not use Comodo Internet...
page itself leads...
thnks u
Are you trying to see your photos using a corporate network (for example, the one from your workplace)? If this is the case, you would need to ask your network admin to whitelist Amazon AWS domain address (amazonaws.com) as suggested by @Welvin.
Let us know if this helps.
no network...
sometimes seems to pictures.
JotForm Support
I think you have not tried yet using other browsers aside from Chrome.
Can you also try testing on Firefox or Safari, and IE browser if you can?
Check if the same issue happen when using another browser.
Please inform us of the results.
I tried. problem continues ...
JotForm Support
Can you please share to us the exact URL link of the file you were trying to download that was being blocked ?
Provide to us the steps taken, and we will try to do the same and see if we can replicate the issue.
I assume you were trying to download the uploaded file in your form's View Submissions page.
JotForm Support
I was able to easily download the file using the link you provided.
but unfortunately, I was not using Comodo Security on my computer. So that was the big difference.
I will seek out advice from my colleague who have Comodo implementation.
We will update you about this soon as we can.


text:
Big personalities are often at the core of what makes a business special. Would Virgin be Virgin without Richard Branson's antics? Would Amazon be the same without Jeff Bezos and his signature cackle? Would an Apple product launch be as buzzworthy without Steve Jobs on stage sporting his signature mock turtleneck?
Undeniably, an entrepreneur's character can be an asset in company building. But when you're ready to sell your business, having a big personality may be a liability, unless you can find a way to inject your persona into your business when you're ready to leave.
I asked Rohit Bhargava, the author of Personality Not Included: Why Companies Lose Their Authenticity and How Great Brands Get It Back , how business owners can make sure their personality becomes ingrained in their business.
Here are three of Mr. Bhargava's ideas:
1. Publish your philosophy
Mr. Bhargava works at the advertising agency Ogilvy, where he has seen first-hand how his company's namesake ensured his ideas would stick around long after David Ogilvy himself sold his interest in the business and moved to an estate in Touffou, France.
Mr. Ogilvy, who died at his French retreat in 1999 at the age of 88, published his philosophies in Confessions of an Advertising Man in 1976 and later in Ogilvy on Advertising in 1985. These books are still essential reading for new Ogilvy employees to ensure they understand the personality of both Mr. Ogilvy the man and Ogilvy the company.
2. Create physical reminders
One of Mr. Bhargava's favourite ideas is a famous Ogilvy quote: "If each of us hires people who are smaller than we are, we shall become a company of dwarfs. But if each of us hires people who are bigger than we are, we shall become a company of giants."
To reinforce the idea, Mr. Ogilvy bought a set of Russian matryoshka (nesting) dolls, which can be opened up to reveal dolls of ever decreasing size. Those dolls can still be found in Ogilvy offices as a reminder for managers to hire people who are smarter than they are.
3. Celebrate your backstory
When James Dyson developed a bagless vacuum that didn't lose suction, he went to visit all of the vacuum manufacturers of the day and asked if they would license the technology. All refused, in part because they liked the ongoing revenue stream of selling bags.
Not content to let a better solution languish, Mr. Dyson spent 15 years developing prototypes on his own before he started his company to bring the vacuum to market.
Among Dyson employees, the backstory of the company and Mr. Dyson's 15-year odyssey became legend. To keep it alive, Mr. Dyson created the James Dyson Design Awards, which celebrate new industrial designers and their commitment to finding better solutions for life's problems.
What are you doing to make sure your personality will live on in your company after you're gone? It could be the key to getting the best price for your business when you're ready to sell.
Special to The Globe and Mail
John Warrillow is a writer, speaker and angel investor in a number of start-up companies. He writes a blog about building a valuable – sellable – company.Report Typo/Error
Follow @JohnWarrillowon Twitter:


text:
(this is my UK SW account I have another one for my work in Israel)
I do volunteer work for a non profit charity in Israel, my predecessor bought some (10 maybe?) licences for Vista, and I think they havent been used.
If I want to declare I dont agree with the licence agreement (I assume they are still in shrinkwrap can I get these refunded? They will never been used as we just have XP and a few Macs.
as the title suggests I dont know if the licence rules are different in Israel, to UK or US.
2 Replies
Mar 12, 2010 at 9:58 UTC
There's going to be some licensing differences depending on the laws of each country. Generally, with software, it's a "You bought it, it's yours." policy.
You could talk to the vendor they were purchased through. If you still have the purchase receipt they may allow you to return it. I wouldn't hold my breath though.
Mar 30, 2010 at 1:04 UTC
Another option might be to just re-sell it if they weren't used.
Your best bet is to talk to Microsoft and see what their policy is on it. Chances are they may say there's not much they can do, but at least that way you hear it from the horse's mouth.
This discussion has been inactive for over a year.
You may get a better answer to your question by starting a new discussion.