text:
|Publication number||US7445887 B2|
|Application number||US 11/326,689|
|Publication date||4 Nov 2008|
|Filing date||6 Jan 2006|
|Priority date||7 Jan 2005|
|Also published as||EP1833980A2, EP1833980A4, US20060154320, US20090068694, WO2006074444A2, WO2006074444A3|
|Publication number||11326689, 326689, US 7445887 B2, US 7445887B2, US-B2-7445887, US7445887 B2, US7445887B2|
|Inventors||Robert Zuk, Sae Choo, Weilei Ma, Krista Witte|
|Original Assignee||Fortebio, Inc.|
|Export Citation||BiBTeX, EndNote, RefMan|
|Patent Citations (54), Non-Patent Citations (33), Referenced by (4), Classifications (9), Legal Events (5)|
|External Links: USPTO, USPTO Assignment, Espacenet|
The present application claims the benefit of U.S. Provisional Application Ser. No. 60/645,153, filed Jan. 19, 2005 and U.S. Provisional Application Ser. No. 60/642,454, filed Jan. 7, 2005, both of which are incorporated herein by reference in their entirety for all purposes.
1. Field of the Invention
The invention relates to interferometry-based methods and compositions useful for measuring enzyme activity.
2. Description of the Related Art
Enzymes represent a broad class of proteins that catalyze biochemical reactions and have many therapeutic and industrial applications. Often in the course of development and manufacture of enzyme products it is necessary to measure the activity of the enzyme. A simple enzyme activity method, preferably label free thereby avoiding perturbation of the enzyme/substrate interaction, would find wide application. In the development and manufacture of enzyme or enzyme inhibitor based products for therapeutic or industrial applications, it is critical to monitor the activity of the enzyme throughout the process. Enzyme assays typically require labeling the substrate in such a way that the enzyme acting on the substrate produces a detectable change in signal. Labeled enzyme substrates are often not commercially available, in which case their synthesis can be complex. For companies developing a multitude of enzyme products, the implementation of activity methods that are simple and easy to perform in research & development and manufacturing environments becomes a major task. The need for labeled substrates adds to the time, expense and inconvenience of enzyme activity measurements. Specific activity measurements also require quantifying the amount of enzyme present in a sample. Quantitation, as by, e.g., enzyme-linked immunosorbant (ELISA)-based assays are also adds to the time and expense of specific activity measurements and requires additional sample. The present invention addresses these and other shortcomings of the prior art by providing simple, fiber based, real-time enzyme activity assays, capable of providing specific activity measurements, suitable for low-volume samples, that are highly multiplexable and in some embodiments can be carried out using unlabeled substrates.
The present invention is defined by the following claims, and nothing in this section should be taken as a limitation on those claims. Disclosed herein are assemblies, kits, and methods for assaying enzyme activity using fiber-based interferometry. In one embodiment, the assay comprises providing an optical element coupled to a light source via an optical fiber and the element includes proximal and distal reflecting surfaces separated by at least 50 nm. A layer of enzyme substrate molecules is positioned so that interference between beams reflected from the proximal and distal reflecting surfaces varies as an enzyme reacts with the substrate. The reflected beams are coupled into the optical fiber. The element is exposed to an enzyme and a change is detected in the interference between the reflected beams. The detected change is indicative of enzyme activity.
In still another embodiment, a layer of analyte binding molecules substituted in the optical element for the layer of enzyme substrate molecules. Interference between beams reflected from the proximal and distal reflecting surfaces varies as an enzyme reacts with the substrate and the acted-upon substrate or portion thereof binds to the analyte binding molecules. In preferred embodiments, the analyte binding molecules comprise an antibody, an antibody fragment, a single chain Fv molecule (“scFv”), an avidin, a streptavidin, or a biotin.
In another embodiment, a semi-permeable membrane is placed between the optical element and the assay solution. In another embodiment, the substrate is coupled to a support such as a microtitre well or a bead.
In yet another embodiment, a similar, second element is provided that includes a layer of molecules that specifically binds to the enzyme. The second element is exposed to an enzyme (either at the same time or at a different time as the first element is exposed) and a change is detected in the interference between the reflected beams. The change is indicative of enzyme concentration or amount. This is useful for carrying out specific activity measurements. In preferred embodiments, the enzyme-binding molecules comprise an anti-enzyme antibody, an antibody fragment or an scFv molecule.
These and other features, aspects, and advantages of the present invention will become better understood with regard to the following description, and accompanying drawings, where:
Briefly, and as described in more detail below, described herein are assemblies, kits and methods for assaying enzyme activity using fiber-based interferometry.
Several features of the current approach should be noted. Measurements can be carried out using extremely small sample volumes (e.g., nL). Measurements can be carried out in vivo or in vitro. In some embodiments, measurements can be carried out on unlabeled substrates while in other embodiments, substrates include a moiety to allow capture by the assembly. In preferred embodiments, the moiety is one member of a binding pair such as, e.g., avidin, streptavidin, biotin, a hapten, an antibody, antibody fragment, an scFv, or a lectin, and the optical element comprises the complementary member of the pair. In these embodiments, the same type of optical element (i.e., carrying one member of the binding pair) can be used in a wide variety of enzyme assays provided the substrate includes the other member of the pair.
Advantages of this approach are numerous. Because the invention provides for fiber-based interferometry measurements, it is sensitive, capable of being highly multiplexed, and easily adapted for specific activity measurements by including a module for measuring enzyme amount.
The invention is useful for measuring enzyme activity in any context for which such activity measurements are useful including, e.g., for discovery, modification, optimization, production, etc. of enzymes or enzyme inhibitors. The invention may be practiced using any type of enzyme such as, e.g., hydrolases, glycosylases, esterases, and transferases, or inhibitors of such enzymes.
Terms used in the claims and specification are defined as set forth below unless otherwise specified.
The term “in vivo” refers to processes that occur in a living organism.
Abbreviations used in this application include the following: dsDNA—double-stranded DNA; dNTPs—deoxynucleotide triphosphates; B-ATP—biotinylated-ATP; PEG—polyethylene glycol, PMSF—phenylmethylsulfonyl fluoride.
It must be noted that, as used in the specification and the appended claims, the singular forms “a,” “an” and “the” include plural referents unless the context clearly dictates otherwise.
Assemblies of the invention include biosensor tips adapted for coupling to a bio-layer interferometer and carrying a layer of substrate or an analyte binding molecule. The analyte binding molecule may be by way of example and not limitation, a member of a binding pair such as, e.g., avidin, streptavidin, biotin, a hapten, an antibody, an antibody fragment, an scFv, or a lectin.
Kits of the invention include a glass fiber adapted for coupling to a bio-layer interferometer, reagents and instructions for derivatizing the glass fiber with a substrate layer or an analyte binding molecular layer, optionally reagents and instructions for optically activating an end of the glass fiber and packaging.
In general, methods of the invention are practiced using assemblies and apparatus, including a bio-layer interferometry (BLI) sensor, such as those described in co-owned U.S. Non-provisional application Ser. No. 10/981,901, filed Nov. 4, 2004, for Hong Tan, et al., entitled “Fiber-Optic Assay Apparatus Based on Phase-Shift Intepferometry,” the contents of which are herein incorporated by reference in their entirety.
In brief, the sensor is prepared by optically activating one end of a glass fiber. The activation steps include buffing the fiber surface, coating the surface with Ta2O5 followed by coating with an SiO2 layer, and cleaning, and immobilization of enzyme substrate or one member of a binding pair by passive adsorption and/or covalent attachment.
Included within the scope of the invention are two broad and general formats for assaying enzyme activity. In the first format, substrate is immobilized on a surface of a bio-layer interferometry (BLI) sensor. In the second format, the BLI sensor includes a surface having a molecule capable of binding substrate. In the second format, a semi-permeable membrane optionally is included between the BLI sensor and the assay solution, or the substrate is bound to a support such as a microtitre well or a bead. These embodiments are particularly useful with hydrolases to prevent or slow binding of full-length substrate to the BLI sensor. In the second format, information about enzyme activity can be derived both from kinetic and steady state components of the interference signal.
A Bio-Layer Interferometry (BLI) sensor is capable of measuring sub nanometer changes in the thickness of its optical layer detection surface. Analysis of biological samples is possible by designing assay formats where biomolecules bind at the sensor surface and change the optical layer thickness. The magnitude of the optical layer thickness change is proportional to the mass or molecular weight of the binding molecule. The Bio-Layer Interferometer can be configured to have substrate immobilized to the sensor surface to measure enzymes whose activity creates a change in the substrate molecular weight, either increasing or diminishing the molecular weight, to produce a corresponding change in the optical layer thickness.
The invention is broadly applicable to enzyme activity measurements, including by way of example but not limitation, measurements of hydrolases (including proteases), glycosylases, esterases, transferases (including nucleotide transferases and phosphotransferases). These are considered in greater detail below.
Below are examples of specific embodiments for carrying out the present invention. The examples are offered for illustrative purposes only, and are not intended to limit the scope of the present invention in any way. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperatures, etc.), but some experimental error and deviation should, of course, be allowed for. Unless otherwise specified, procedures are carried out at room temperature (typically 20-23 degrees Celsius).
The practice of the present invention will employ, unless otherwise indicated, conventional methods of protein chemistry, biochemistry, recombinant DNA techniques and pharmacology, within the skill of the art. Such techniques are explained fully in the literature. See, e.g., T. E. Creighton, Proteins: Structures and Molecular Properties (W.H. Freeman and Company, 1993); A. L. Lehninger, Biochemistry (Worth Publishers, Inc., current addition); Sambrook, et al., Molecular Cloning: A Laboratory Manual (2nd Edition, 1989); Methods In Enzymology (S. Colowick and N. Kaplan eds., Academic Press, Inc.); Remington's Pharmaceutical Sciences, 18th Edition (Easton, Pa.: Mack Publishing Company, 1990); Carey and Sundberg Advanced Organic Chemistry 3rd Ed. (Plenum Press) Vols A and B(1992).
The minimum molecular weight of a binding molecule that BLI can detect is illustrated in
The small minimum molecular size detection limit of BLI makes it possible to apply BLI to a large number of enzymes. The following are by way of example, but not limitation, enzyme classes whose activities can be measured in accordance with the present invention, and specific examples of such measurements.
Hydrolyases are enzymes that catalyze cleavage of C—O, C—N, C—C or phosphoric anhydride bonds.
Subgroup 1: Proteases (Enzymes Acting on Peptide Bonds) Immobilized Substrate Format
This format features the protease substrate immobilized to the surface of the BLI glass fiber sensor using methods described in co-owned U.S. Non-provisional application Ser. No. 10/981,901, filed Nov. 4, 2004, for Hong Tan, et al., entitled “Fiber-Optic Assay Apparatus Based on Phase-Shift Interferometry,” (incorporated herein by reference) and below. The fiber is immersed in an enzyme-containing sample, and monitored for changes in optical layer thickness.
The basic assay protocol is to incubate a Bio-Layer Interferometer (BLI) sensor on which has been immobilized an enzyme substrate in an enzyme-containing solution enzyme. The amount of the substrate depletion is quantified by, e.g., a change in optical phase shift using the Bio-Layer Interferometry (BLI) technique fully described in co-owned and pending U.S. Non-provisional application Ser. No. 10/981,901, filed Nov. 4, 2004, for Hong Tan, et al., entitled “Fiber-Optic Assay Apparatus Based on Phase-Shift Interferometry,” incorporated herein by reference. The change in, e.g., optical phase shift is proportional to the amount of enzyme activity in the solution because hydrolytic activity is estimated by measuring the depletion of substrate from the BLI sensor resulting from the enzyme activity.
Subtilisin Activity Measurements on Immobilized Casein Substrate
An optical signal baseline was established by immersing the optically-activated end of a fiber sensor tip in PBS and monitoring the optical signal using interferometry methods and instrumentation described in co-owned U.S. Non-provisional application Ser. No. 10/981,901, filed Nov. 4, 2004, for Hong Tan, et al, entitled “Fiber-Optic Assay Apparatus Based on Phase-Shift Interferometry,” (incorporated herein by reference in its entirety). Next the fiber was coated with Poly-D-Lysine by incubating the tip in a 0.5 mg/mL Poly-D-Lysine solution (in PBS, pH 7.4) for 15 minutes. Unbound Poly-D-Lysine was rinsed by incubating the tips in PBS for 10 minutes.
The fiber was coated with a layer of casein [Sigma Chemical Company, St Louis, Mo.] by incubating the tip in a 50 μg/mL casein solution (50 mM Na Phosphate, 150 mM NaCl, pH 7) for 15 minutes. Unbound casein was rinsed by incubating the tip in PBS for 10 minutes.
Fibers with immobilized casein were incubated in various concentrations (1, 10, 50 μg/mL) of subtilisin [Sigma Chemical Company, St Louis, Mo.] solution (in 50 mM Na Phosphate, 150 mM NaCl, pH 7). Each of the procedures described above was carried out while monitoring the optical signal.
Results and Discussion
Effect of Protease Inhibitor on Enzymatic Activity
Fiber sensor tips were prepared and coated with casein as described above. One fiber was incubated in 50 μg/mL of subtilisin solution (in 50 mM Na Phosphate, 150 mM NaCl, pH 7).
Other fibers were incubated in a pre-mixed and pre-incubated solution of 50 μg/mL of subtilisin and 3 mM PMSF [Sigma Chemical Company, St Louis, Mo.] (the mixture was incubated at 37° C. for 20 minutes) solution (50 mM Na Phosphate, 150 mM NaCl, pH 7). Each of the procedures described above was carried out while monitoring the optical signal.
Result and Discussion
Traces obtained from a fiber incubated in subtilisin solution show the expected depletion of casein from the tip surface (
Substrate Capture Format
The substrate capture format entails digestion of the substrate by a protease in liquid phase followed by binding of the substrate to the surface of the BLI sensor. The binding of the substrate is designed so that proteolytic cleavage of the substrate produces a detectable to change in the optical layer thickness. In one preferred embodiment, a streptavidin/biotin binding pair is used to effect substrate capture.
Trypsin activity measurements are made using the substrate capture format and cytochrome C as a substrate. Cytochrome C is about 12 kDaltons in molecular weight and includes has 8 trypsin cleavage sites. A standard biotin-NHS derivative [Pierce Biotechnology, Rockford Ill.] is used to tag the cytochrome C. Coupling conditions employ standard phosphate buffered saline (PBS) pH 7 buffer. Biotin-NHS is mixed with cytochrome C at a molar coupling ratio of 5 to 1 (biotin to cytochrome C) typically resulting in one biotin substitution per cytochrome C molecule. Streptavidin-coated BLI sensors are prepared from 0.6 mm diameter glass fibers with a tantalum oxide layer of about 20 nm and a silicon dioxide layer of about 700 nm are dipped in a PBS solution containing 0.5 mg/ml of poly-D-lysine as described above. After 15 minutes at room temperature, the fibers are washed in PBS then immersed in a 1 mg/ml solution of bovine serum albumin (BSA) labeled with N-succinimidiyl 3-(2-pyridyldithio) propionate (SPDP) [Pierce Biotechnology, Rockford Ill.] and incubated for 20 minutes followed by a wash step with PBS. The fibers are then immersed in a solution of 50 mM dithiolthreitol for 30 minutes at room temperature. After a wash in PBS, the fibers are placed in the solution containing 20 μg/ml of streptavidin labeled with SMCC and incubated for 60 minutes, followed a by PBS wash. The fibers are stored in PBS until use.
The basic assay for protease activity using the substrate capture format entails adding to a biotinylated cytochrome C solution, in the range of about 1 μg/ml to 1 mg/ml, a trypsin sample, typically at a 1% wt./wt. ratio. After a digestion time period, a streptavidin coated sensor is placed in the enzyme/substrate mixture for substrate capture. To avoid trypsin acting upon the streptavidin, the enzyme can be inactivated either by snap boiling the substrate mixture or adding a protease inhibitor, such as aprotinin, just prior to adding the glass fiber. The BLI sensor can assess the change in substrate size by a measuring the optical layer thickness upon binding the biotinylated peptide.
Subgroup 2: Glycosylases (Enzymes that Hydrolyse O or S or N Glycosyl Bonds) Immobilized Substrate Format
The following example is described for dextranase, but the same methods can be applied to other polysaccharide processing enzymes such as amylase and cellulase. Dextran is linked to a protein coated BLI sensor by initially reacting the dextran with sodium periodate to introduce reactive aldehyde groups on the dextran polymer. A protein coated BLI sensor (such as prepared according to the methods described in the example above) is then immersed in the dextran solution. The aldehyde groups on the dextran form bonds with free amino groups on the protein resulting in a BLI sensor with a dextran as the final layer. Measurement of dextranase activity is possible when the dextran coated sensor is immersed in a dextranase containing sample. Hydrolytic activity of the dextranase reduces the molecular size of the immobilized dextran, which is detected as a thinner optical layer.
Substrate Capture Format
Measurement of glycosylases in the substrate capture format is accomplished by biotinylating the polysaccharide substrate. In the dextranase example, dextran is reacted with sodium periodate to generate aldehyde groups followed by the addition of large molar excess of a bis-amine, such as ethylenediamine. One amino group of the bis-amine couples to the aldehyde on the dextran leaving the second amine free for coupling to biotin-NHS in a subsequent reaction. The molar coupling ratio of the biotin-NHS reaction is selected to yield about one biotin substitution per dextran. The assay for dextranase activity in the substrate capture format is similar to the protease second example where the biotinylated dextran substrate is incubated with the dextranase sample followed by binding of the biotinylated dextran to a streptavidin coated BLI sensor. Dextranase activity creates smaller dextran fragments which are measured as a thinner optical layer.
Subgroup 3: Esterases (Enzymes that Act on Ester Bonds)
Some examples of esterases include nucleases (RNase, DNase, etc.), alkaline phosphatase, acid phosphatase, and serine/threonine phosphatase. DNase I is used as an example of measuring esterase activity. Since DNase I cleaves at all 4 bases in oligonucleotides as small as three bases in length, dsDNA is prepared by a commercial vendor using standard DNA synthesis techniques having a length of about 30-40 base pairs with a biotin group at one of the terminal ends. The biotinylated dsDNA is bound to a streptavidin coated BLI sensor. The sensor is then immersed in sample containing DNase I in the buffer: 10 mM Tris pH 7.5, 2.5 MgCl2, 0.5 nM CaCl2 and the decrease in optical layer thickness is monitored. The alternative substrate capture format is performed by mixing the biotin labeled dsDNA with the DNase I in liquid phase before the binding of the substrate to the streptavidin coated sensor.
Transferases are enzymes that catalyze the transfer of methyl, glycosyl, or phospho groups to other compounds. In contrast to the hydrolytic enzymes, the transferases increase the molecular size of the substrate and their activity is detected by the BLI sensor as an increase in optical layer thickness.
Subgroup 1: Nucleotide Transferases
Nucleotides transferases catalyze the incorporation of nucleotides into DNA or RNA polymers.
An alternative approach to measure activity of nucleotide transferases is based on hapten incorporation as shown in
Subgroup 2: Phosphotransferases
Phosphotransferases catalyze the transfer of phosphate groups to hydroxyl containing compounds, typically peptides with tyrosine, serine, or threonine residues. There are over 100 commercially monoclonal antibodies that bind to a phosphorylated amino acid or specific sequences encompassing a phosphorlyated amino acid. Many assays of kinase activity have been reported and are commercially available utilizing antibodies to phosphorylated peptide substrates.
While the invention has been particularly shown and described with reference to a preferred embodiment and various alternate embodiments, it will be understood by persons skilled in the relevant art that various changes in form and details can be made therein without departing from the spirit and scope of the invention.
All references, issued patents and patent applications cited within the body of the instant specification are hereby incorporated by reference in their entirety, for all purposes.
|Cited Patent||Filing date||Publication date||Applicant||Title|
|US4830451||5 Mar 1986||16 May 1989||American Telephone And Telegraph Company||Technique and apparatus for fabricating a fiber Fabry-Perot etalon|
|US5301001||12 Feb 1992||5 Apr 1994||Center For Innovative Technology||Extrinsic fiber optic displacement sensors and displacement sensing systems|
|US5359405||6 Nov 1992||25 Oct 1994||Martin Marietta Corporation||Hybrid fiber optic sensor including a lead out optical fiber having a remote reflective end|
|US5422970||3 Dec 1993||6 Jun 1995||Micron-Optics, Inc.||Temperature compensated fiber Fabry-Perot filters|
|US5425039||24 Feb 1994||13 Jun 1995||Micron Optics, Inc.||Single-frequency fiber Fabry-Perot micro lasers|
|US5452087||4 Nov 1993||19 Sep 1995||The Texas A & M University System||Method and apparatus for measuring pressure with embedded non-intrusive fiber optics|
|US5528367||9 Sep 1994||18 Jun 1996||The United States Of America As Represented By The Secretary Of The Navy||In-line fiber etalon strain sensor|
|US5594819||26 Jul 1995||14 Jan 1997||Electric Power Research Institute||Field-mountable fiber optic sensors for long term strain monitoring in hostile environments|
|US5606170||3 Feb 1995||25 Feb 1997||Research International, Inc.||Multifunctional sensor system|
|US5629214||31 May 1995||13 May 1997||Biostar, Inc.||Methods for forming an optical device for detecting the presence or amount of an analyte|
|US5631171||31 May 1995||20 May 1997||Biostar, Inc.||Method and instrument for detection of change of thickness or refractive index for a thin film substrate|
|US5647038||30 Aug 1995||8 Jul 1997||Hughes Aircraft Company||Narrow bandwidth Bragg grating reflector for use in an optical waveguide|
|US5682237||26 May 1995||28 Oct 1997||McDonnell Douglas||Fiber strain sensor and system including one intrinsic and one extrinsic fabry-perot interferometer|
|US5701193||21 Feb 1996||23 Dec 1997||Ascom Tech Ag||Optical reflection modulator|
|US5732169||22 Dec 1995||24 Mar 1998||Alcatel Submarcom||Filter for guided light, and an optical link including the filter|
|US5804453||9 Feb 1996||8 Sep 1998||Duan-Jun Chen||Fiber optic direct-sensing bioprobe using a phase-tracking approach|
|US5869835||27 May 1997||9 Feb 1999||Udd; Eric||Asymmetric fiber optic grating sensor|
|US5982959||11 Jul 1998||9 Nov 1999||Hopenfeld; Joram||Coated fiber optic sensor for the detection of substances|
|US6055080||5 Jun 1997||25 Apr 2000||Deutsche Forschungsanstalt Fur Luft-Und Raumfahrt E.V.||Optical microphone|
|US6078706||22 Sep 1998||20 Jun 2000||The United States Of America As Represented By The Secretary Of The Navy||Quasi-static fiber pressure sensor|
|US6139797||20 Aug 1998||31 Oct 2000||Suzuki Motor Corporation||Immunoassay apparatus|
|US6241397||16 Apr 1999||5 Jun 2001||Micron Optics, Inc.||Cascaded fixed and tunable multiple-fiber Fabry Perot filters and fixed and tunable ferrule alignment fixtures for fiber Fabry Perot filters|
|US6244214||22 Dec 1999||12 Jun 2001||Embrex, Inc.||Concurrent in ovo injection and detection method and apparatus|
|US6248539||30 Oct 1997||19 Jun 2001||The Scripps Research Institute||Porous semiconductor-based optical interferometric sensor|
|US6254830||5 Nov 1999||3 Jul 2001||The Board Of Governors For Higher Education, State Of Rhode Island And Providence Plantations||Magnetic focusing immunosensor for the detection of pathogens|
|US6275628||15 Nov 1999||14 Aug 2001||Luna Innovations, Inc.||Single-ended long period grating optical device|
|US6277651||9 Jul 1998||21 Aug 2001||Calspan Srl Corporation||Diode laser electrochemical sensor for detecting chemical and biological analytes|
|US6281976||8 Apr 1998||28 Aug 2001||The Texas A&M University System||Fiber optic fiber Fabry-Perot interferometer diaphragm sensor and method of measurement|
|US6445838||29 Sep 2000||3 Sep 2002||Corning Incorporated||Tunable optical component|
|US6496618||14 Feb 2002||17 Dec 2002||Varian, Inc.||Fiber-optic channel selecting apparatus|
|US6539136||24 Mar 1999||25 Mar 2003||Nauchny Tsentr Volokonnoi Optiki Pri Institute Obschei Fiziki Rossiiskoi Adademii Nauk||Fiber-optic pressure sensor, variants and method for producing a resilient membrane|
|US6571639||1 Mar 1999||3 Jun 2003||Luna Innovations, Inc.||Fiber optic system|
|US6590665||16 Mar 2001||8 Jul 2003||Institut National D'optique||Optical sensing devices|
|US6611334||14 Feb 2002||26 Aug 2003||Varian, Inc.||Sample analysis system with fiber optics and related method|
|US6661520||22 May 2002||9 Dec 2003||National Taiwan University||Sensor system of surface plasmon resonance (SPR) and measuring method thereof|
|US6671055||13 Apr 2000||30 Dec 2003||Luna Innovations, Inc.||Interferometric sensors utilizing bulk sensing mediums extrinsic to the input/output optical fiber|
|US6687011||22 Dec 1999||3 Feb 2004||Korea Advanced Institute Science And Technology||Transmission-type extrinsic fabry-perot interferometric optical fiber sensor|
|US6720177||28 Feb 2001||13 Apr 2004||The Regents Of The University Of California||Porous semiconductor-based optical interferometric sensor|
|US6744939||20 May 2002||1 Jun 2004||Fitel Usa Corp.||Polarization maintaining optical fiber connector and method of tuning (PM connector)|
|US6870624||30 Oct 2001||22 Mar 2005||Coho Holdings Llc||Optical wavelength resonant device for chemical sensing|
|US6870630 *||9 Oct 2003||22 Mar 2005||Novartis Ag||Sensor platform, apparatus incorporating the platform, and process using the same|
|US7158225||23 Jan 2004||2 Jan 2007||Kaiser Optical Systems||Multi-channel, self-calibrating fiber-coupled raman spectrometers including diagnostic and safety features|
|US20010048072||16 Mar 2001||6 Dec 2001||Yves Painchaud||Optical sensing devices|
|US20020127565 *||15 Aug 2001||12 Sep 2002||Sru Biosystems, Llc||Label-free high-throughput optical technique for detecting biomolecular interactions|
|US20030112443||18 Oct 2002||19 Jun 2003||Optomed. As||Optical sensing of measurands|
|US20030215791 *||23 Oct 2002||20 Nov 2003||Applied Spectral Imaging Ltd.||Method of and system for multiplexed analysis by spectral imaging|
|US20040022475||17 Apr 2003||5 Feb 2004||Luna Innovations, Inc.||Biosensors having enhanced environmental sensitivity|
|US20040151626 *||23 Oct 2001||5 Aug 2004||Brian Cunningham||Label-free high-throughput optical technique for detecting biomolecular interactions|
|US20040186359||9 Jul 2002||23 Sep 2004||Beaudoin Stephen P.||Afinity biosensor for monitoring biological process|
|US20050254062 *||4 Nov 2004||17 Nov 2005||Fortebio, Inc.||Fiber-optic assay apparatus based on phase-shift interferometry|
|US20060050358||15 Oct 2003||9 Mar 2006||Joel Bigman||Electrochemically controlled optical devices|
|US20070070356 *||12 Jun 2006||29 Mar 2007||Hong Tan||Fiber-Optic Assay Apparatus Based on Phase-Shift Interferometry|
|USH1813||19 Nov 1993||2 Nov 1999||Kersey; Alan D.||Spectrally-selective fiber transmission filter system|
|WO2005047854A2 *||5 Nov 2004||26 May 2005||Fortebio, Inc.||Fiber-optic assay apparatus based on phase-shift interferometry|
|1||Boiarski, A. et al., "Integrated-Optic Biosensor", SPIE, Fiber Optic Sensors in Medical Diagnostics, vol. 1886, 1993, pp. 15-26.|
|2||*||Brecht A. et al. Recent Developments in Optical Transducers for Chemical or Biochemical Applications. Sensors and Actuators B 38-39, 1-7, 1997.|
|3||Brecht, A. et al., "Direct Monitoring of Antigen-Antibody Interactions by Spectral Interferometry", Sensors and Actuators, vol. B5, 1992, pp. 96-100.|
|4||Brecht, A. et al., "Interferometric Immunoassay in an FIA-System: a Sensitive and Rapid Approach in Label-free Immunosensing", Biosensors & Bioelectronics, vol. 8, 1993, pp. 387-392.|
|5||Brecht, A. et al., "Recent Developments in Optical Transducers for Chemical or Biochemical Applications," Sensors and Actuators B, Jan. 1997, pp. 1-7, vol. 3-39, Elsevier Science S.A., Lausanne, CH.|
|6||Brecht, A. et al., "Theoretical and Experimental Detectivity of the RIFS-transducer in Affinity-sensing", Biosensors 94, The Third World Congress on Biosensors: Abstracts, Oral Session, Jun. 2, 1994, p. 68.|
|7||Cao, L. et al., "Detection of Yersinia Pestis Fraction 1 Antigen With a Fiber Optic Biosensor", Journal of Clinical Microbiology, vol. 33, No. 2, Feb. 1994, pp. 336-341.|
|8||Christensen, D. et al., "Analysis of Excitation and Collection Geometries for Plannar Waveguide Immunosensors," SPIE vol. 1886, Fiber Optic Sensors in Medical Diagnostics, 1993, pp. 2-8.|
|9||Davies, R. et al., "An Optical Biosensor System for Molecular Interaction Studies", American Biotechnology Laboratory, Jul. 1993.|
|10||Elster, J. L. et al., "Optical Fiber Extrinsic Fabry-Perot Interferometric (EFPI)-Based Biosensors," Proceedings of the SPIE, 2000, pp. 105-112, vol. 3911.|
|11||European Examination Report, EP 04800761.1, Nov. 23, 2007, 5 pages.|
|12||European Supplementary Search Report, EP 04800761, Mar. 19, 2007, 4 pages.|
|13||Fabricius, N. et al., "A Gas Sensor Based on an Integrated Optical Mach-Zehnder Interferometer", Sensors and Actyators, vol. B7, 1992, pp. 672-676.|
|14||Gauglitz, G. et al., "Observation of Spectral Interferences for the Determination of Volume and Surface Effects of Thin Films", Analytical Biochemistry, vol. 341, 1991, pp. 279-283.|
|15||Hogg, D. et al., "Development of a Fiber Fabry-Perot Strain Gauge", SPIE vol. 1588, Fiber Optic Smart Structures and Skins IV, 1991, pp. 300-307.|
|16||Jorgenson, R. et al. "A Novel Surface Plasmon Resonance Based Fiber Optic Sensor Applied to Biochemical Sensing", SPIE, vol. 1886, 1993, pp. 35-48.|
|17||Kimoshita, Y. et al., "Sensing of Herbicide Residues Using Surface Plasmon Resonance Technique", The Third World Congress on Biosensors: Abstracts, 1994, p. 257.|
|18||Lim, S. et al., Application of an Interferometric Biosensor Chip to Biomonitoring an Endocrine Disruptor. Biotechnology and Bioprocess Engineering. 2004, vol. 9, No. 2, pp. 118-126.|
|19||Lin, C-J., et al., "A Novel in Vitro and in Situ Immunoassay Biosensor Based on Fiber Optic Fabry-Perot Interferometry," Proceedings of the SPIE, The International Society for Optical Engineering, 2004, pp. 304-307, vol. 5502, No. 1.|
|20||Lukosz, W., et al., "Output grating Coulers on Planar Optical Waveguides as Direct Immunosensors", Biosensors & Bioelectronics, vol. 6, 1991, pp. 227-232.|
|21||Lundstrom, I., et al. "Immunosensors Based on Surface Plasmon Resonance", The Third World Congress on Biosensors: Abstracts, 1991, p. 91.|
|22||Office Action, Chinese Application No. 200480031823.5, Jul. 20, 2007, 16 pages.|
|23||Ogert et al., "Detection of Clostridium Botulinum Toxin A Using a Fiber Optic-Based Biosensor", Analytical Biochemistry, vol. 205, 1992, pp. 306-312.|
|24||PCT International Search Report and Written Opinion, Aug. 4, 2006, PCT/US06/00700.|
|25||PCT International Search Report and Written Opinion, PCT/US06/22964, Jun. 1, 2007, 9 pages.|
|26||PCT International Search Report and Written Opinion; PCT/US04/36830, Sep. 6, 2005, 9 pages.|
|27||Reagent Reservoirs with Control Wells, V&P Scientific, Inc., 2006, [online] [Retrieved on Sep. 6, 2006] Retrieved from the Internet <URL:http://www.vp-scientific.com/reagent<SUB>-</SUB>reservoirs<SUB>-</SUB>with<SUB>-</SUB>control<SUB>-</SUB>.htm>.|
|28||Rudraraju, S. et al., "Acoustic Wave Propagation in Composite Materials: an Experimental Study", SPIE, vol. 2191, Jul. 1994, pp. 487,493.|
|29||Schneider, I. et al., "Herbicide Detection Using Reaction, Centers Integrated Into Liposomes Binding to Grating Couplers", The Third World Congress on Biosensors: Abstracts, 1994, p. 271.|
|30||Tiefenthaler, K., "Grating Couplers as Label-free Biochemical Waveguide Sensors", Biosensors & Bioelectronics, vol. 8, No. 7-8, pp. xxxv-xxxvii, no date given.|
|31||Weber, A. et al., "Fiber-optic Fluorimetry in Biosensors: Comparison Between Evanescent Wave Generation and Distal-face Generation of Fluorescent Light", Biosensors & Bioelectronics, vol. 7, 1992, pp. 193-197.|
|32||Yang Y. et al., "Direct Monitoring of Antigen-Antibody Interactions by Optical Fiber Bioprobe," Proceedings of the SPIE, Jun. 8, 2003, pp. 431-436, vol. 5254, No. 1.|
|33||Yang, Y. et al., "Study of Optical Fiber Biosensor Based on White-Light Interferometry," Journal of Xi'An Jiaotong University, Sep. 2003, pp. 914-916, 988, vol. 37, No. 9, Xian, CN.|
|Citing Patent||Filing date||Publication date||Applicant||Title|
|US8512950||20 Jul 2011||20 Aug 2013||Saint Louis University||Biolayer interferometry measurement of biological targets|
|US8647588||4 May 2011||11 Feb 2014||Pall Corporation||Tip tray assembly for optical sensors|
|US20100093106 *||14 Sep 2007||15 Apr 2010||Fortebio, Inc.||Amine-Reactive Biosensor|
|US20110236911 *||18 Mar 2011||29 Sep 2011||Fortebio, Inc.||Precipitating Substrate for Bio-Layer Interferometry|
|U.S. Classification||435/4, 356/901|
|Cooperative Classification||G01N33/573, C12Q1/00, G01N33/54373|
|European Classification||C12Q1/00, G01N33/573, G01N33/543K2|
|6 Jan 2006||AS||Assignment|
Owner name: FORTEBIO, INC., CALIFORNIA
Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:ZUK, ROBERT;CHOO, SAE;MA, WEILEI;AND OTHERS;REEL/FRAME:017452/0524
Effective date: 20050413
|6 Jun 2012||FPAY||Fee payment|
Year of fee payment: 4
|6 Jun 2012||SULP||Surcharge for late payment|
|8 Jun 2012||AS||Assignment|
Owner name: PALL FORTEBIO CORP., CALIFORNIA
Free format text: MERGER;ASSIGNORS:FORTEBIO, INC.;PROJECT PHOENIX ACQUISITION CORP.;REEL/FRAME:028345/0275
Effective date: 20120301
|4 May 2016||FPAY||Fee payment|
Year of fee payment: 8

domain: biology


Are you having trouble booking a campsite during the busy summer months? A pair of North Vancouver entrepreneurs think they might have the solution.
The new website, called “Campertunity,” is being called the “Airbnb of camping,” and it works along similar lines.
Hosts who own a piece of private land can list it through the site. They choose the price, with Campertunity taking 10 per cent. So far, it’s attracted about 16 properties — with nightly prices ranging from $6 to $50.
LISTEN: The ‘Airbnb of camping?’
The concept takes aim at what has been a chronic problem for would-be campers: hot demand for provincial campgrounds.
The province changed its Discover Camping reservation system last year to create a booking window in which spots become available four months in advance, rather than all at once.
That change was in response to complaints of overbooking and campsite scalping, but many people are still having trouble landing a spot on desirable dates.
Speaking with CKNW’s Lynda Steele Show, Campertunity co-founder Guita Yazdani said that frustration was a driving force behind the new site.
“When you go to book a campground, you’re dealing with stressful bookings, resales — booking a campsite in B.C. is like buying concert tickets, there’s this mad rush for it,” she said.
“I can’t even plan my afternoon, so I don’t know how I can plan my summer in the winter.”
WATCH: Extreme camping tactics: What people are doing to get a spot
She said she and her business partner Nora Lozano fleshed the idea out after camping on someone’s piece of private property near Pemberton.
A few weeks later, they camped again — this time in a provincial campground — and something clicked.
“We woke up and it was just to the sound of RVs going by us, families, kids walking by, going to the bathroom, and I remember thinking you know, if I close my eyes right now, I would think I’m in the city,” she said.
“That’s the difference, I felt,” Yazdani said. “I thought, ‘Wow, private land, and then provincial campground. Why isn’t camping on private land a thing?'”
WATCH: Province changes camping reservation system
Campertunity isn’t the first local digital platform to take aim at the high demand for B.C. campgrounds.
Last year, a B.C.-based entrepreneur launched an app that lets would-be campers snag popular campsites that have had a last minute cancellation — for a fee.
The province says it is spending $22.9 million over five years to add 1,900 new campsites to take pressure off the system.
But it says by April 30 this year, there had already been 100,000 reservations made through the Discover Camping website, up 30 per cent over last year.
To discourage scalping, the province has also made it so that campers can not change the name on reservations.


text:
Performing Songwriters Live In The Lounge
Come on out to "Nashville Songwriters Live In The Lounge" with your host Uncle Rock!
7 p.m. - 10 p.m. there will be songwriters in the round playing all original songs!
The Nashville PieHoles will play around 9 p.m. and looking forward to it!
Four Points has a full menu, bar selection and a lovely atmosphere! Free parking too!! No cover

domain: entertainment


text:
By Matt Hardigree, Jalopnik Chrysler’s big reveal at the Detroit auto show was the 2013 Dodge Dart, but this shiny jellybean was sitting at the far end of the display. The automaker calls it the Chrysler 700C. We’re calling it …
All posts tagged ‘Chrysler’
- 5:30 AM
- 6:00 AM
After 37 years, Chrysler brings back the Dart nameplate with its first cool small car since the Dodge Neon.
- 8:30 AM
Our vita just got a little more dolce as Fiat confirms the Abarth 500 pocket rocket is headed here next year.
- 2:00 PM
Chrysler plans to introduce compressed natural gas-powered vehicles in 2017 and could raid the Fiat parts bin to make it happen. “The technology is very actively being worked on,” Bob Lee, Chrysler’s vice president for engine and electrified propulsion systems, …
- 9:00 AM
An analysis of online responses to automakers’ advertising campaigns has found that Chrysler’s “Born Of Fire” commercial featuring rapper Eminem continues to strike a chord online. Alterian, a company that tracks and analyzes social media, found that the online buzz …

domain: news


text:
Statue in garden
B907/0009 Rights Managed
530 pixels on longest edge, unwatermarked
Please login to use the price calculator
Release details: Model release not required. Property release not required.
Keywords: art, bed, blue-purple, border, borders, botanical, botany, delphinium, delphiniums, flower, flowering, flowers, garden, garden scene, horticultural, horticulture, pink, plant, sculpture, statuary, statue, statue in garden, statue with delphiniums 43, statues, stone, stonework, topiary
Licence fees: A licence fee will be charged for any media (low or high resolution) used in your project.

domain: artistic


text:
Six Flavors of Crypto Influencers
I have gotten a lot of information and education from crypto influencers over the past year+. Some of the picks have wound up paying handsome rewards and others have fallen flat. Regardless, I’ve noticed that many of these influencers share certain characteristics. In this article, I highlight some of the character traits that many crypto influencers fall under.

domain: finance


Ceramacast™ 900 is a zirconium-silicate based, calcium-aluminate bonded, chemical-setting castable ceramic ideal for casting small parts and fixtures, and for potting high temperature electrical components such as infrared heaters, cartridge heaters, gas igniters, halogen lamps, and temperature sensors for applications to 2800 ºF (1535 ºC).
Ceramacast™ 900 exhibits exceptional electrical and mechanical properties. The dielectric strength and volume resistivity are 125 volts/mil and 109 ohm-cm, respectively. The compressive strength is 8,000 psi and the coefficient of thermal expansion (CTE) is 2.8 (4.0) ppm/ºF (ºC).
Ceramacast™ 900 is supplied as a two-part, powder and liquid, system that is mixed in a ratio of 100 parts powder to 15 parts liquid by weight. It is a rapid-setting compound with a pot life of 20 minutes to assist with quick turn-around. A full cure is achieved by step curing for 2–3 hours each at 200 ºF and 250 ºF, respectively. Cured product exhibits a porosity of less than 5% and shrinkage of less than 1.0% at 1000 ºF.
Ceramacast™ 900 is supplied from stock in quart, gallon and five gallon kits. It is one of a series of Ceramacast™ products that have been developed for ultra high temperature electrical assembly and casting applications. Please contact Aremco’s Technical Sales Department for more information about Ceramacast™ 900, or read more.


text:
Travel companies must move past getting people from A to B, but think about collecting information to build a 360° view of the traveler to create a tailored and memorable experience – right from the moment they are inspired to travel, to the time they return home from their trip.
With the provision of unlimited, uncapped Internet to guests fast becoming the biggest benefit for hotels in future.
Chatbots and artificial intelligence are some of the new technologies to watch out for and are already being used by travel companies to respond to the changing needs of travelers, says Adams.
“Some companies have introduced virtual reality to allow travelers to ‘try before they fly’,” she says. “Amadeus will soon launch the Traveller Notifier tool, which sends important notifications to the traveller’s mobile phone, such as electronic ticket issuance, flight delay, boarding gate assignment and boarding gate change. Virtual reality will also continue to gain moment in 2018 in the travel industry.”
VR does something means consumers can be transported anywhere. For example, Virgin Holidays are using headsets and a 3D camera to build a 360-degrees holiday experience that people can use to explore destinations. You can’t do these types of immersive experiences with any other tech.
The big opportunity for VR will be helping consumers make decision about where to travel, according to analysts at tourism market research firm Phocuswright.
“It can be especially valuable for destinations that may not have a top-tier attraction with a lot of name recognition, but has great natural cultural attractions that can give travelers confidence that this is the place to go,” Douglas Quinby, vice president of research at Phocuswright, told CNBC via email.
An example of this comes in the form of virtual tours, which will allow travelers to experience a holiday destination before booking their trip.
A Uganda tour company, Matoke Tours, a niche African travel operator, launched a virtual travel brochure: an app featuring 360degree videos of six experiences in Uganda, where users will come face-to-face with a gorilla or go up in a hot air balloon.
“This app enables us to convey the intensity and emotion of the travel experience before the journey has even started,” said Wim Kok, director of Matoke Tours Uganda, in a press release. “Travelers are then better able to decide which excursions they want to book.”
There are already several apps offering this kind of experience; Ascape is a free app for the iPhone offering 360degree videos and virtual tours of locations such as Berlin and Botswana.
South African Tourism promotes destination in the UK with VR activity. They took over two bars in Manchester and London last year, inviting customers to watch a short film on the virtual reality headset Oculus Rift. The ‘five-minute holiday’ film transports the viewer on to the side of Table Mountain, where they experience what it’s like to abseil down the sheer cliff, with 360-degree views of the sea behind and city in the distance.
Viewers are then absorbed into other adventurous scenes, such as shark cage-diving in Cape Town, kite-surfing, paragliding and feeding elephants, before relaxing into a bar at Johannesburg’s Neighbourgoods Market
Juan Herrada, UK manager of marketing and communications for South African Tourism Board, said “We want to surprise the UK travelling public and Oculus Rift is the ideal partner for us to reach a young, tech savvy market. This is the first time Oculus Rift and binaural sound have been combined to create a unique, immersive holiday experience for the UK public.”
People trying Oculus Rift will be blown away. It’s a great use of creative technology by a tourist board and we are confident it will encourage more people to visit South Africa.

domain: travel


text:
Masai Mara Wildbeest Migration Safari
WHAT IS INCLUDED IN THE PRICES
a) Transport in a safari vehicle for 3 days
b) 2 nights’ full board accommodation
c) Meals on Full board for 2 nights (meals are Buffet)
d) Unlimited game drives for 3 days
6 Days Masai Mara - Lake Nakuru - Amboseli Safari
Safari Highlights: 6 Days Budget or Luxury Lodge safari to Masai Mara, lake Nakuru and Amboseli National park. This safari features the open plains of the famous Masai Mara Game Reserve for unparalleled game viewing in one of Africa's best game parks, famous for its annual migration of Wildebeests and Zebras, the pink waters of Lake Nakuru home to millions of lesser and greater flamingoes. Amboseli game viewing with the majestic Mount Kilimanjaro at the backdrop. Amboseli is a compact game park, abundant in wildlife, which makes for some excellent game viewing.
4 Days Masai Mara - Lake Nakuru Safari
Safari Highlights: Explore two of Kenya's top game parks - Masai Mara and Lake Nakuru National Parks - on thrilling game viewing drives over 4 adventure-filled days. Masai Mara is a large game reserve measuring 1,510 kilometers square in south western Kenya contiguous with Serengeti National park in Tanzania, it is famous for its exceptional population of lions, leopards, cheetahs and its annual migration of zebra and wildebeest which occur from July to October hence known as the great migration while Lake Nakuru national park found in the central Kenya is one of the Rift
4 Days Lakes Bogoria and Lake Nakuru - Crescent Island - Hells Gate Safari
Day 01: Nairobi – Lake Bogoria: This is a shallow lake found in the Rift Valley, covering an area of 30 square kilometer, with a depth of 9 meters and home to thousands of greater and lesser flamingoes. Other attractions are the HOT SPRINGS and GEYSERS (with the Hot Springs boiling hot), gazelles, greater kudu, and impala. As you plan your bird watching safari in Kenya, you can look forward to recording your sightings of crombec warblers, swifts, gobar goshawks, cape wigeons, starlings, hornbills, ostriches and many other raptor, terrestrial and wading species.
3 Days Masai Mara Safari
Safari Highlights: This is a short safari featuring Masai Mara National Reserve - one of the world's finest wilderness areas. The safari departs Nairobi in the morning and involves a drive down to the floor of the Great Rift Valley, making a stop at view point to see the Great Rift Valley escarpment and the awesome sight of the valley and the features there in. Maasai Mara game reserve supports the largest diversity of species in Africa with close visit to homeland of the Masai people, a proud cattle-herding tribe.
3 Days Amboseli Safari
Tour Summary/ Features: This 3 day safari takes you through Southern Kenya towards the Amboseli National Park, which lies in the shadows of Mount Kilimanjaro, Africa's highest mountain. Amboseli is known for its rich wildlife, particularly its prolific elephant population. This Kenya Safari will be at Amboseli National Park which is Kenya's international biosphere reserve. Amboseli in Masai language means "place of water" and contemporary Maasai people's culture and indigenous lifestyle.

domain: geography


Todd Thurber, PE
Master of Business Administration, University of Houston
Bachelor of Science, Civil Engineering, University of Texas
Todd Thurber, PE, has nearly three decades of transportation project experience in the Houston region and throughout the state. Areas of expertise include planning, development, and design for expansion and reconstruction of freeway corridors traversing and linking urban centers in Texas. His experience encompasses new corridor development, highway rehabilitation and expansions, complex urban interchanges, feasibility studies, toll feasibility and tolling operations, transit projects including HOV and HOT lanes, and traffic studies for TxDOT, Harris County Toll Road Authority, Fort Bend County Toll Road Authority, Houston METRO, and the Grand Parkway Association. As project manager on the planning and final design of urban freeways/direct connect interchanges, he has completed over $1B worth of projects. He continues to promote improved mobility in our region by volunteering his expertise to ACEC Houston on the Public Affairs Committee and as subcommittee chair for the joint ACEC/TxDOT/HGAC Planning Process Workshop, and the Fort Bend Chamber of Commerce, where he was a board member and chair of the Infrastructure Division. He has been recognized for his efforts, being selected as 1998’s SAME Young Engineer of the Year and the 2015 ASCE Texas Section Citizen Engineer Award. He is also active in the community with organizations such as the ARC of Fort Bend County.


text: It’s finally here! Release day for One Dance with a Duke, the first installment in an all-new trilogy. (And if you haven’t watched the Stud Club Trilogy video yet, you really should!) To celebrate the book’s release, I’m hosting a very special giveaway. Details here.


Groundbreaking outboard engine parts rely on reinforced nylons
Rugged, beautifully finished cowling components for Mercury Marine's revolutionary Verado outboard boat engine swept the annual Product Design Competition sponsored by The Society of the Plastics Industry's Structural Plastics Division in March, c...
Rugged, beautifully finished cowling components for Mercury Marine’s revolutionary Verado outboard boat engine swept the annual Product Design Competition sponsored by The Society of the Plastics Industry’s Structural Plastics Division in March, capturing four awards for innovation and groundbreaking design. The cowl consists of six parts injection molded from DuPont high-performance plastics by Bemis Manufacturing Co.
The cowl fulfills structural, protective and aesthetic functions while reducing weight and cost compared with Mercury’s alternative materials.
The top cowl is believed to be the largest component ever injection molded from glass-reinforced nylon. Made of a 33% glass reinforced DuPont Zytel nylon, it is shaped like a giant scoop measuring 851 mm (33.5 in.) front to back, 582 mm (22.9 in.) across and up to 417 mm (16.4 in.) deep. It weighs just 11.3 pounds (5.13 kg), far less than a metal or a thermoset plastic housing.
The rear cowl, and a structural rib that is adhesive-bonded to it, are molded from the same Zytel resin. The front cowl is molded from DuPont Minlon mineral/glass reinforced nylon for added flatness. The lower cowling, or chaps, consists of two mating parts molded from DuPont Surlyn Reflection Series supergloss alloy for superior gloss without paint.
These components meet several demanding requirements:
They mate precisely to prevent water from entering the engine compartment.
They provide improved noise/vibration/harshness properties, making Verado the quietest, lowest vibration engine in its class
They provide structural strength and impact resistance.
Stand up to temperatures up to 200F in the upper cowl.
The cowl parts made of Zytel and Minlon nylon resins are finished in a three-coat system providing first-pass acceptance yields of 95% or better at each stage, according to Mercury Marine.
The lower cowl molded from DuPont Surlyn Reflection Series supergloss alloy has high-gloss finish and integral color. It incurs no painting costs.
DuPont Canada Inc. 905-821-5953


text:
Treatment of upper respiratory tract infections in cats typically involves well-managed supportive care and possibly administration of oral medications. The goals of therapy are to control secondary bacterial infections, maintain the cat’s weight and hydration and keep the cat as comfortable – for as long - as possible.
Cats with upper respiratory tract infections should be isolated from other cats for at least three to four weeks, to prevent contagion. All bowls, bedding, crates, toys and other things that an infected cat has come into contact with should be thoroughly and regularly washed with a 1:32 dilute solution of bleach-to-water. People who touch an infected cat should be especially attentive to good hygiene, including washing their hands with soap and warm water and changing and laundering their clothes and shoes on a regular basis.
Most affected cats can be treated at home. This is preferable to in-hospital care, at least from the cat’s perspective. At-home supportive care for cats with upper respiratory infections is very similar to how we care for people with “the flu.” The cat should be in a calm, warm and quiet environment that promotes rest and relaxation. A home vaporizer will improve humidification and help to keep the cat’s nasal passages moist. Some authorities recommend a cool steam vaporizer rather than a warm vaporizer. Placing the cat on a dry surface in a steamy bathroom for 10 to 15 minutes several times daily is also helpful. Many owners bring their cat into the bathroom with them while they shower or bathe.
Supportive treatments for upper respiratory infections in cats also include keeping the cat in a confined, stress free and comfortable in-home environment. The cat should be in a room that has a comfortable temperature. Free access to fresh water is important, and the cat’s diet should be highly palatable. The cat’s eyes and nose should be routinely cleaned if the cat is experiencing eye or nose discharge. Cats with respiratory illness usually have runny noses and goopy eyes. Owners should gently cleanse their cat’s eyes and nose with warm water on a soft tissue or cotton ball as frequently as necessary to remove the infectious secretions. They should dispose of those tissues or cotton balls in a place that is reliably inaccessible to other animals.
Cats with upper respiratory tract infections often have difficulty eating, because they cannot smell their food. In addition, when they develop oral ulcers, affected cats frequently refuse to eat or drink due to pain. This can cause dramatic weight loss and profound weakness. Owners should feed highly palatable foods that have a strong, pungent smell, such as canned tuna, canned mackerel or strained meat baby food. Some cats will need to be force-fed either by hand or through a tube. High-calorie nutritional paste supplements are also available and can be recommended or provided by a veterinarian. If a cat becomes dehydrated during its illness, subcutaneous or intravenous fluids can be administered by the owner or by a veterinarian. Water can also be given using a needleless syringe or eye dropper.
Broad-spectrum oral antibiotics can be used to treat bacterial causes of upper respiratory tract infections in cats. Decongestant drops may be recommended to shrink swollen nasal membranes. Topical ointments and artificial tears are available for cats whose eyes are affected. Topical or systemic antiviral drugs may also be appropriate. Appetite stimulants can help cats that are anorexic, and pain medications (analgesics) can help cats with oral sores. The amino acid, L-lysine, can be helpful to reduce the severity of respiratory infections in cats that test positive for feline herpesvirus.
Most cats with upper respiratory tract infections have a good to very good prognosis, if they receive prompt and effective treatment. Young kittens, and cats with especially virulent viral infections, have a more guarded prognosis. Infected adults rarely die from upper respiratory tract infections. Unfortunately, the death rate in very young kittens can approach fifty percent.

domain: pet


text:
UltraPure BSA is a non-acetylated bovine serum albumin (BSA) pure enough to use when DNA or RNA integrity is critical. One tube containing 50 mg is provided, at a concentration of 50 mg/mL. BSA has many uses as a carrier protein and as a stabilizing agent in enzymatic reactions. In northern, Southern, and dot blot hybridizations, BSA is also used as a blocking agent. It is recommended in buffers for nick translation, polymerase reactions, and ligations. BSA is also a common additive for PCR amplifications, footprinting, and gel shift assays. In restriction digests, BSA has been shown to enhance enzyme activity. The molecular weight is 68 kD. UltraPure BSA is rigorously tested for DNase, RNase, endonuclease, and protease activity.
For Research Use Only. Not for use in diagnostic procedures.

domain: chemistry


text:
You will love wearing these southwestern dream catcher dangle earrings.
The dream catcher earrings are made with antiqued copper feature blue beads around the center.
The copper wire wrapped dream catcher earrings are lightweight and easy to wear all day and right into the night.
You can wear these unique dangle earrings for casual wear, work or a night out.
These artisan made earrings would also make a great gift for a family member, a friend or even a co-worker.
The dream catcher earrings are 7/8th inches wide and are 2 1/8th inches long (not including the nickel free ear wires).
The measurements can be seen in the last 2 photo.
Each piece of my wire wrapped jewelry is handmade, so no two pieces will ever be alike or "perfect".
Slight irregularities in each piece should not be considered as them having a defect.
I take great care and attention in wrapping my jewelry in order to get the best wire wrap possible.
**Wire wrapping uses pliable wire for each piece. It will retain it's shape, however if you try to bend it or it gets squashed, it will bend out of shape.**

domain: design


text: Angela Tong is a designer, instructor, mother and food lover living in New Jersey. She designs knitting, crochet and weaving patterns for numerous magazines, books and yarn companies. She is a certified knitting instructor through the Craft Yarn Council. She enjoys teaching knitting and weaving classes online for Craftsy.com and Interweave and in person at Vogue Knitting Live, Maryland Sheep & Wool Festival, yarn stores, guilds and fiber retreats. In 2019, she became a brand ambassador for Mirrix Looms. Follow her daily crafting adventures on Instagram @atongdesigns. She has never met a craft that she didn’t like.


text:
Body Shaming Out; Chubby Is The New In: Find Out How Miss Universe Canada Made Waves At The Miss Universe 2016
Body shaming is not much of a new issue for Miss Universe 2016 candidate Siera Bearchell. The 23-year-old Canadian beauty has not only endured body shaming from trolls as she has also triumphed over them by making it to the recently concluded Miss Universe 2016 competition final 13 cut. Bearchell was found to have graced the Miss Universe stage on January 29 in Manila, Philippines, after taking on her haters in a public Instagram post earlier this month. She said that she even feels happier and more confident than she ever has.
Body Shaming: How It Affects A Person?
According to reports revealed by New Mic, on January 17, Bearchell has allegedly posted a photo of herself in a swimsuit, along with a message about body positivity, as response to all of her detractors. It was found that long before, a significant number of Psychologists have already done a lot of research on this, and the evidence is very clear. It was found that body shaming, in all forms does not motivate people, but makes them feel terrible about themselves and actually causes them to eat more and gain more weight. Furthermore, the said series of research shows that the stigma on body shaming against overweight people causes major psychological harm and makes the problem worse.
Miss Universe Canada: On Redefining Beauty Standards
Meanwhile, as per The Sydney Morning Herald, during the contestants' press junket, Miss Canada, Siera Bearchell was left speechless after being asked of how she feels being so much larger as compared to the other delegates. However, instead of reacting negatively, Bearchell has instead responded through her social media account saying she "feels great" to be part of a movement to redefine beauty standards. Ultimately, until the actual pageant, the 23-year-old Siera Bearchell has managed to land a top-9 finish despite a lot of people previously doubting her.
Elon Musk Reveals First Images Of Tesla Model 3 Production Unit On Twitter
Elon Musk showed off the very first Tesla Model 3 unit on Twitter and Instagram.
Avoid Scams On Social Media With These Helpful Tips
Social media network is a place full of opportunities especially for people devising scams so pay attention and find out to spot scams.
Elon Musk Is Working On A Consensual Telepathy; Latest Details Inside
Elon Musk has confirmed that Neuralink is real and that he is serious about the technology. The firm will soon develop a “neural lace” technology that would involve implanting tiny brain electrodes.
Starbucks Unicorn Frappucino Is A Total Instagram Bait, Find Out Why
The Starbucks Unicorn Frappuccino, a limited-edition drink was released this week and became a viral trend all over the social media, most especially on Instagram.
Robots Recruited To Teach Kids In China And Canada
In China and Canada, AI-driven robots have been recruited to help teach kids learn things in the classroom. The well-researched machines have been proving to be significant additions in the lives of young children.
MORE IN ITECHPOST
'Tokyo Ghoul' Season 3 Delayed By Its Live-Action Movie? Ken Kenaki Returns With Another Identity?
"Tokyo Ghoul" season 3's release is delayed and there are countless speculations as to why it is being pushed back. Reports claim that the show is to give way to "One Punch Man" and "My Hero Acdemia" but new rumors say that it's lack of materials and storyline as well as its live-action movie are causing the delay.


A tool that detect file added/modified and transfer the file automatically.
What is this ?
After a file is created/modified, the file will be transferred automatically between two folder (Internal Storage or External SD Card), with below features:
– Supports sub folder
– Supports overriding file
– Show notification after file moved
– Scan media after file moved
– Set detection delay time
– Allow multiple task (premium feature)
– Allow turning on/off task (premium feature)
– Backup and restore task setting (premium feature)
– No Ads (premium feature)
Why you need this ?
1. Some apps (camera, phone screenshot, …) only can save file in internal storage, so this app can help to transfer the file to external SD card
2. Some apps (NAS file sync app, …) only able to access certain SD card location, so this app can help to transfer the file to other
3. anything that you can think
Q & A
Q1: This app is not working on my Android 8 phone ?
A1: Most Android 8+ systems prevent applications from running in the background, please check system settings and allow this application to run in the background. Please select schedule option “Instant (With Notification)” in the setting, if the notification is not displayed, it means the application was killed by the system.
*** Important ***
1. Only supports Android 5.0 or above
2. Please create a testing task(test folder with test file) for your first use.
3. This application has been extensively tested. Despite that, we will not be responsible for any data loss or damage.
► 4.1.0 – 4.1.4
1. New implementation of folder selection
2. App optimized
3. Bug fixed
*** Please backup before update (Premium) ***
☆Premium features unlocked
☆Ads Layout removed/Ad activity Removed
☆Remove default .source name of java files;
☆Debug Info Removed
This app has no advertisements
Downloads : Premium Features Unlocked


On the Dubai Creek bank is Dubai Creek Harbour, a waterfront neighbourhood. The neighbourhood provides resorts, a yacht club, a marina, retail spaces, and entertainment venues, among other mixed-use developments with a view of the ocean. Emaar Properties created Dubai Creek Harbour.
The residential neighborhood of Dubai Creek Harbour is known for its dynamic environment and a wide range of property types, including townhouses, flats, and villas. For residents who are interested in waterfront living, the area offers a variety of residential options, including high-rise penthouses that boast panoramic views of the creek and the Burj Khalifa.
This waterfront community is easily accessible from Downtown Dubai, making it a convenient option for those who live here. Those who are looking for a more convenient location may consider renting a unit in one of the many condos and apartments in the area.
Apartments in Dubai Creek Harbour offer uninterrupted views of the city skyline and feature luxurious amenities, including gyms and swimming pools. Residents also have access to ample parking.
The waterfront community of Dubai Creek Harbour is home to a diverse range of real estate offerings. The district features a range of luxury residences, luxury boutiques, offices, and other commercial entities, surrounded by a lush oasis of greenery. The area has many attractions, including Dubai Creek Tower, which is the city’s tallest tower. Aeon & Trisl is the premier real estate agency in the city.
The skyline of Dubai is spectacular. At the top of the tallest tower, the city’s most popular skyline can be seen. Located near the city’s mangroves, the community offers an exceptional range of properties, many of which are freehold. Residents can also enjoy access to the many freehold ownership rights. The area has all of the amenities and services that a city of this size and location demands.
The residential area of Dubai Creek Harbour is centered on the Island District, which features 13 high-rise projects and two hotels. The first completed project is the Dubai Creek Residences, which comprises six towers – three on either side of the creek. The second is the Creek Gate, which consists of two towers. The next two projects are the Creek Edge and the Creek Rise, as well as the new Dubai Marina. In addition, residents can also explore the Ras Al Khor bird sanctuary and Creek Island’s central park.
Those looking to live in the area should consider the area’s proximity to schools. The nearest elementary school is Repton School, which is approximately 15 minutes away. Other elementary schools are located in Al Hamra, while the Repton school is just a 15-minute drive away. There are many amenities in Dubai Creek Harbour that are sure to please any resident. The neighborhood is a fantastic addition to the city. Whether you’re looking for a quiet place to spend your time, this area of the city is an excellent option.


text:
- Condition: Used - Excellent
- Brand: WELCH ALLYN
- Type: Autorefractor
- Model: Spot
- MPN: RE0WAVS100SBFG
Welch Allyn Spot Vision Screener - VS100 Handheld Autorefractor
The Spot Vision Screener is an easy to use autorefractor that accurately identifies refractive errors and ocular misalignment in children and adults. Ideal for adults with small pupils (measures 3mm and greater), non-verbal patients, children, and patients with disabilities. The portable handheld autorefractor allows testing refractive errors easier and more efficient through a non-invasive 3-foot distance. The autorefractor is equipped with a touchscreen display with on-screen instructions to guide the user through the process. Provides an easy-to-use and easy-to-understand solution for eye care professionals.
This is a used model. In like new, perfectly functioning condition.

domain: medical


text:
Whilst every effort is made to ensure the details on this website are correct, the publishers of cannot guarantee the accuracy of the information.
Avoid a wasted journey - Always confirm event details directly with the event's organisers.
Myton Moor: Between the Swale and the Kyle (Grade 2)
Sat 25 Apr 2020
Verges near Myton Gates
Phone: 07583 451825 / 07570 112010Send Email
Additional Event InformationNew for 2020, this delightful 10-mile ride explores the quiet rural area north of York and east of Boroughbridge, between the Rivers Swale and Kyle. The route follows bridleways and tracks, with some roads. The terrain is flat, so this is a good ride if your horse is not particularly fit. We pass through the village of Aldwark, and pause at the New Inn at Tholthorpe for refreshments. The ride is led by Janet Cochrane. Max. 6 riders, start time 10 am, entry fee £30 (includes a drink).


AMSOIL Antifreeze and Coolant is a top-of-the-line product that helps keep your engine running cool and efficiently. It's formulated with a patented organic corrosion inhibitor package that provides superior protection against rust and corrosion, and it's compatible with all types of antifreeze and coolant. AMSOIL Antifreeze and Coolant is also designed to protect against freezing, boil-over, and corrosion in all types of engines, making it an ideal choice for your vehicle.
AMSOIL Coolant Boost is an additive that helps to improve the performance of your coolant system. It is designed to increase the efficiency of your coolant system by reducing the formation of deposits and corrosion. AMSOIL Coolant Boost also contains a corrosion inhibitor that helps to protect your engine from rust and corrosion.
Is AMSOIL Propylene Glycol Antifreeze and Engine Coolant compatible with aluminum intakes, cylinder heads and engine blocks? Yes. AMSOIL Antifreeze and Engine Coolant (ANT) is compatible with all engine and cooling system components. It surpasses all ASTM standards for metallic corrosion to protect metals and provide uncompromised cooling protection.
Does AMSOIL recommend a particular type of water when mixing AMSOIL Antifreeze and Engine Coolant? AMSOIL recommends deionized or distilled water when mixing with AMSOIL Antifreeze and Engine Coolant (ANT). Using deionized or distilled water prevents adding harmful contaminants that can negatively affect the antifreeze product and cooling system equipment.
Is AMSOIL Antifreeze and Engine Coolant silicate and borate free? Yes. AMSOIL Antifreeze and Engine Coolant (ANTPC, ANTHD) contains proprietary poly-organic acid technology that is silicate and borate free.
Become a Preffered Customer Today!
Add a six-month or one-year Preferred Customer membership to your cart and save up to 25 percent on today’s order.


text:
Take advantage of July 4th sales. Set the table in red, white and blue with patriotic accessories.
These items are fun and affordable. A stripe straw makes a drink festive any day of the week.
gingham check napkins/runner, tooth picks, star napkins, straws, image via
About Ivy de Leon
From decorating and home staging to entertaining and gardening, I enjoy bringing traditional style to each project. I love to combine new and vintage market finds to create elevated and inviting spaces for today's modern lifestyle.
This entry was posted in Decorating
and tagged patriortic
, table top
, white and blue
. Bookmark the permalink

domain: hobby


text:
Pig Emoji Coloring Pages.
Use the download button to find out the full image of Pig Emoji Coloring Pages, and download it for a computer. You could also print the picture by clicking the print button above the image.
This Pig Emoji Coloring Pages would make your world more vibrant. Be ready for some coloring enjoyable with complimentary coloring sheet. Surf the broad option of free coloring sheets for children to find educational, animations, nature, animals, Bible coloring books, as well as a lot more.


Service with heart and SOL!
COURTESY CALL| Earlier today, personnel of Philippine Statistics Authority (PSA) XII, PSA South Cotabato, and PSA Sultan Kudarat paid Mayor Solly F. Villalobos a courtesy call. During the visit, the PSA informed the Mayor and the Municipal PhilSys Focal Person about the alleged falsification and possession of fake, falsified, counterfeit, and or altered Philippine Identification Cards in the provinces of South Cotabato and Sultan Kudarat. In response, Mayor Solly commits to help the PSA XII with whatever support it requires during the fact-finding process. (cmpe)


text:
Harry Potter star Emma Watson has distanced herself from franchise author JK Rowling's comments on transgender people.
Rowling has been criticised by former and current co-workers, as well as LGBTQ+ fans and allies alike, for commenting on transgender people by posting a tweet about menstruation: "People who menstruate. I'm sure there used to be a word for those people. Someone help me out. Wumben? Wimpund? Woomud?".
Earlier this week, Rowling said that she had been "empathetic to trans people for decades" and denied that her comments were transphobic. She added: "I respect every trans person's right to live any way that feels authentic and comfortable to them. I'd march with you if you were discriminated against on the basis of being trans."
However, following renewed backlash for Rowling penning a 4,000-word blog post on the issue that ignited yet more condemnations on Wednesday (June 10), Watson tweeted to let her transgender fans know she supports them.
"Trans people are who they say they are and deserve to live their lives without being constantly questioned or told they aren't who they say they are," she tweeted.
"I want my trans followers to know that I and so many other people around the world see you, respect you and love you for who you are."
"Sending love," she added.
Her Harry Potter franchise co-star Daniel Radcliffe previously penned an essay for The Trevor Project to tell fans he was "deeply sorry for the pain" Rowling's words had caused, noting that nearly 80% of transgender and nonbinary youth have been reported to have faced discrimination.
We would encourage anyone who identifies with the topics raised in this article to reach out. Anyone who has witnessed or experienced a hate crime is urged to call the police on 101, Crimestoppers anonymously on 0800 555 111 or visit the True Vision website. In an emergency, always dial 999.
Digital Spy now has a newsletter – sign up to get it sent straight to your inbox.
Looking for more TV recommendations and discussion? Head over to our Facebook Group to see new picks every day, and chat with other readers about what they're watching right now.


text:
Thoresby Hall Two course dinner for twoThoresby Hall - Two course dinner for two
Near Ollerton, Nottingham
Harvester Steak, Slow Cooked Ribs or Chicken Meal with Wine and Unlimited SaladHarvester - Steak, Slow Cooked Ribs or Chicken Meal with Wine and Unlimited Salad
Tom Browns Brasserie Chateaubriand & Bubbly for 2Tom Browns Brasserie - Chateaubriand & Bubbly for 2
Trentside Gunthorpe, Nottingham
The Riverbank Bar and Kitchen Two Course Riverside Meal & Bubbly for 2The Riverbank Bar and Kitchen - Two Course Riverside Meal & Bubbly for 2
Trent Bridge, Nottingham
Quorn Country Hotel Sunday Two Course Lunch with Wine for Two or FourQuorn Country Hotel - Sunday Two Course Lunch with Wine for Two or Four
Leicester Rd, Quorn
The Farndon Boathouse Two Course Meal & Bubbly for 2 on the River TrentThe Farndon Boathouse - Two Course Meal & Bubbly for 2 on the River Trent
Off Wyke Lane Farndon, Newark
The Wick Meal with a Bottle of Prosecco to Share for TwoThe Wick - Meal with a Bottle of Prosecco to Share for Two
Hilton House Hotel Sunday carvery lunch for twoHilton House Hotel - Sunday carvery lunch for two
Diletto Waffle or Crepe with Ice Cream and Hot Drink for Up to FourDiletto - Waffle or Crepe with Ice Cream and Hot Drink for Up to Four
East St, Derby
Ugly Bread Bakery Hot Mezzaluna with Hot DrinkUgly Bread Bakery - Hot Mezzaluna with Hot Drink
Pelham St, Nottingham
Money saving in Nottingham is made easy with Marc and Daina's Restaurant Groupon vouchers from www.dealzippy.co.uk.
DealZippy.co.uk have got 0 Marc and Daina's Restaurant deals running today and in the past we've found 1 Marc and Daina's Restaurant vouchers that have now run out.
Our Nottingham members have saved as much as £26.80 or 52% using deals for Marc and Daina's Restaurant Nottingham from the many UK deal websites we partner with.
Sign up with DealZippy.co.uk today to see how much you could save on things you already buy in Nottingham and be emailed when Groupon or one of the more than 30 alternative deal sites we track holds a Marc and Daina's Restaurant Nottingham deal.

domain: food


text:
Under new regulations set by the UK’s Financial Conduct Authority (FCA), crypto advertisements must now prominently feature explicit risk warnings.
The FCA has directed firms to inform clients, through their ads starting 8 October this year, that they may not be shielded should their investments falter.
Such alerts might suggest to customers that should any issues arise, they should be ready to forfeit their entire investment. The FCA also encourages companies to provide a ‘cooling off’ duration for novice investors.
Companies advertising crypto assets like Bitcoin will be required to offer a delay period for new investors looking to engage in their offerings. Additionally, incentives for referring friends to a company’s products will be prohibited under the new guidelines.
Sheldon Mills, the FCA’s Executive Director of Consumers at Competition, stated in The Guardian, “It’s individuals’ choice to purchase crypto, yet many laments making rash decisions. Our regulations provide people with the necessary time and appropriate risk warnings to make a well-informed decision.”
He further cautioned, “Consumers should remain mindful that the crypto sector is primarily unregulated and high risk. Investors should be ready to lose their entire investment.”
If anyone reads this article found it useful, helpful? Then please subscribe www.share-talk.com or follow SHARE TALK on our Twitter page for future updates.
Terms of Website Use
All information is provided on an as-is basis. Where we allow Bloggers to publish articles on our platform please note these are not our opinions or views and we have no affiliation with the companies mentioned

domain: economics


text:
It’s Tuesday again. That wonderful day when it isn’t Monday and the week is still full of promise. Some of you have noticed that I like to have my interview on Tuesdays. I am in the middle of doing a promotional series of my own publishing house of eTreasurespublishing by interviewing some of the authors there. Today the Marketing Director has given her time to join us. I feel very lucky to be able to feature her. Her name is Patricia Bates. Grab your mocha and sit a spell. You’re in for a treat. Her genre is romance and she has written over eight books, four of which I’ll show here today and four next Tuesday in part two of our interview. (I break it up in two parts because we are all so busy and it’s easier to read this way.) 🙂
Thanks, Patricia. Here’s a start
1- What books are currently published and by who? Can you tell us a little about them?
I have a number of books. All of them are romance books, ranging from 8th Century Ireland to paranormals, contemporaries and m/m titles. Some of them are written under my pen name Elise Whyles and others are under my real name.
A complete list is available on my websites – but here’s an abridged one lol
Master’s Mistress, Love Thy Neighbor, Viscount’s Prize, and Claiming her Dream is through CBG
The Cowgirl’s Christmas is through Beachwalk as well as **The Forsaken Series – a paranormal erotic series featuring mythological creatures and various mythologies
The Preacher’s Outlaw is through Decadent Publishing
**Five Alarm Lust and **Real Good Man are through Liquid Silver Books
Bordering on Love, Sin’s Bite, and **Gillian’s Christmas Wish are all self-pubs. The Books with ** are under Elise Whyles
Phantom’s Pleasure and Firecracker are through Passion in Print
2- Did you go the traditional route? Or consider self publishing any of them? Why? Why not?
Most of the books are actually published through e-publishers though I have self pubbed a few of them due to length, or inability to find a home due to getting rights reversals. When I started my journey a few years ago I never imagined how much I’d learn and what I would like.
3- How long does it take to write a book for you from idea to finished product?
It depends entirely on the length and heat level. If it’s a full length novel say around 45k – and I don’t have a deadline staring me in the face – I can punch it out in three months…if I have a deadline I need to get it in by – as little as three weeks. If it’s shorter (something I don’t like to do because as someone who knows me – I’m a word hog) it can take as little as a week.
4- You are the promotional arm for eTreasures publishing are you not? Can you tell us a little bit about this job and what you do?
Its one of the hats I wear here at eTreasures, yes. Promoting a publishing house is a lot different then a single author brand. I’m constantly looking for ways to reach readers of all the genres, using the social media I have available can be tricky but its something I really enjoy. I book things like blog tours, readers groups on fb, and do the newsletter for the house. Looking for review sites, organizing contests, and sales are all part of it. I do work closely with the authors as well, to ensure that they get a chance to promote because marketing can be scary for someone who hasn’t done it before.
5— How can people get in touch with you? What is the name of your blog and tell us what you blog about?
If people want to get in touch with me they can check out both my websites www.patbatesromances.com orwww.elisewhyles.com. As well they can check out my blogs – http://ofinkandquille.blogspot.com andhttp://charactersandcoffee.blogspot.com
Of Ink and Quille is my blog about a little bit of everything, my writing, my journey, things that I feel strongly about and a spotlight for other authors. Characters and Coffee is my character interview blog..
Reading has been such a large part of my life. I cut my literary teeth on such author’s as Louis L’Amour, Nora Roberts and Janet Dailey. For me it wasn’t such a jump from reading the wonderful tales these authors’s spun to imagining my own.
Soon I was writing poetry, short stories and by junior high I’d written my first full length romance novel. Since then I’ve taken my love of history and my passion for writing and combined them into what I hope will continue to prove a successful career.
Thanks so much, Patricia, for letting me feature you on my blog. It has been a pleasure! Next week we’ll have part two of this interview with Patricia Bates, author and promotional editor of eTreasurespublishing. It is the very same house that published my children’s picture book, “Annie’s Special Day.”
I bet you didn’t know this house was so diverse, did you? Do you think you might read any of these romances? Why or why not? And we just adore questions so ask away and we’ll meet by the old tree called comments. :)
- Clarbojahn Presents! Author Cindy Cromer! Part One. (clarbojahn.wordpress.com)
- Clarbojahn Presents! Author Cindy Cromer! Part Two. (clarbojahn.wordpress.com)
- Clarbojahn Presents! Author Dixiane Hallaj with New Book “It’s Just Lola”! (clarbojahn.wordpress.com)
- Clarbojahn Presents! Author John Doody! Part One. (clarbojahn.wordpress.com)
- Clarbojahn Presents! Author John Doody! Part Two. (clarbojahn.wordpress.com)
- About My Life Saver: Stormi Johnson (clarbojahn.wordpress.com)


Latest Fanny Palli Petralia Stories
By DEREK GATOPOULOS ATHENS, Greece - A Greek cruise ship that struck a volcanic reef and forced the evacuation of hundreds of tourists sank on Friday, 15 hours after it began taking on water off the coast of a Mediterranean island. A Frenchman and his daughter were missing, officials said. Passengers on Thursday climbed down rope ladders to coast guard boats below in a three-hour rescue that involved Greece's military, commercial ships and local fishermen from the island of Santorini....
- Good cheer; viands.
- To revel; to feast.


Press Release Summary:
Entek Enwatch PX Online Surveillance System connects to continuous protection monitoring system outputs, allowing buffered eddy current probe data acquisition without signal-conditioning device. System simultaneoulsy samples paired channels to improve diagnostics. Acquired data can be displayed in orbit and shaft centerline analysis plots, as well as spectra, time waveforms, and order vectors via plant asset management software package over existing Ethernet LAN.
Original Press Release:
Rockwell Automation Expands Continuous Protection-Monitoring System Diagnostics with New Online Surveillance Instrument
Two-channel capability of Entek Enwatch PX delivers quicker and more accurate diagnosis of potential equipment problems
MILWAUKEE, May 28, 2002 - Responding to manufacturers' need to
cost-effectively and quickly pinpoint potential equipment problems before they proliferate, complete automation leader Rockwell Automation has expanded the power of the Entek Enwatch Online Surveillance System. The addition of two-channel simultaneous data acquisition offered in the Enwatch PX surveillance instrument enhances the diagnostic capabilities of
existing continuous monitoring systems, helping to ensure operational data reaches plant management before equipment fails.
"Identifying machinery problems at an early state enables companies to make intelligent decisions regarding equipment availability," said George Hofer, market specialist, Rockwell Automation. "By using existing sensors and buffered output data from installed protection monitoring systems, Enwatch PX cost-effectively captures critical equipment information in a timely fashion."
The Enwatch PX surveillance instrument connects easily to continuous protection monitoring system outputs. This allows buffered eddy current probe data acquisition without a costly signal-conditioning device. And since the Enwatch PX integrates directly with existing continuous monitoring systems, sensor installation costs are minimal.
The Enwatch PX delivers improved diagnostics by simultaneously sampling paired channels. Acquired data can be displayed in orbit and shaft centerline analysis plots, as well as typical plots like spectra, time waveforms and order vectors, via a plant asset management software package over an existing Ethernet local area network (LAN). A total of eight pairs of two channels can be multiplexed from a single Enwatch PX unit or inputs can be configured for up to 16 individual proximity probe inputs from a single machine. This critical data can be used in a predictive capacity to improve equipment downtime planning.
Integrated with additional Rockwell Software packages, such as the Emonitor Odyssey machinery information system or Emonitor Enshare plant asset management software, Enwatch PX provides a fully automated data collection program. Using these software programs, end-users can see a complete picture of equipment condition - the first step toward achieving a successful e-manufacturing environment.
Rockwell Automation (NYSE: ROK) is a world-leading provider of industrial automation power, control and information solutions that help customers meet their manufacturing productivity objectives. The company brings together leading brands in industrial automation for Complete Automation solutions, including Allen-Bradley controls and engineered services and Rockwell Software factory management software, Dodge mechanical power transmission products, and Reliance Electric motors and drives. The company also is a leading provider of contact management technologies and applications that help companies more efficiently manage interaction with their own customers.
Headquartered in Milwaukee, Wis., USA, the company employs approximately 23,000 people at more than 450 locations serving customers in more than 80 countries.
All reader inquiries should be directed to the Rockwell Automation Response Center, 10701 Hampshire Avenue South, Bloomington, Minn., 55438, 800.223.5354, Ext. 1527.
Enwatch, Emonitor, Odyssey, Enshare, Complete Automation, Dodge, Reliance Electric, Allen-Bradley, and Rockwell Software are trademarks of Rockwell Automation.


text:
Hilary Duff reveals that she follows Gwyneth Paltrow’s diet and admits to “starving off her hunger.” However, she clarifies that it is not to an extreme point and that she allows herself to indulge at times. Duff believes that finding a balance and listening to one’s body is important for maintaining a healthy lifestyle.
Hilary Duff revealed that she follows Gwyneth Paltrow’s diet and “starves off her hunger.”
Actress and singer Hilary Duff recently revealed in an interview that she follows Gwyneth Paltrow’s diet. During an appearance on “The Dr. Oz Show,” Duff admitted that she “starves off” her hunger by sticking to Paltrow’s strict eating plan, which includes only a handful of foods.
Although Duff admits that the diet is difficult to maintain, she also says that it allows her to stay fit and healthy. She credits Paltrow with helping her adopt a healthier lifestyle and says that she continues to follow the actress’s advice when it comes to diet and exercise.
Paltrow’s diet, which she calls the “Cleanse,” involves cutting out most processed foods, dairy, gluten, and sugar. Instead, followers are encouraged to eat lots of vegetables, lean proteins, and healthy fats. While some critics have slammed the diet as too restrictive and potentially dangerous, others say that it’s a great way to jumpstart a healthy eating plan.
For Duff, it seems that the diet has been a success. She says that she feels great and that she’s able to maintain her energy throughout the day. While not everyone may want to follow Paltrow’s strict eating guidelines, it’s clear that the actress’s healthy habits are having a positive impact on those around her, including Duff.


text:
I’m feeling pretty ghetto in my new dripping Chanel tee;
I customised this tee by cutting the sleeves off low, this looks great with a lace bra.
Add a chunky chain for an edgy look;
Showing off the Eiffel T tatt;
I’m wearing: Cocaine and Caviar customised tee, River Island peg leg trousers and Converse.


Third Member of Laguna Pueblo Sentenced for Conviction Arising Out of Drive-By Shooting of Acoma Pueblo Home
ALBUQUERQUE – Joseph Edward Lucero, 25, of Laguna Pueblo, was sentenced this morning to 74 months in federal prison followed by two years of supervised release for his assault conviction arising out of the drive-by shooting of an Acoma Pueblo home.
Lucero is the third of four members of Laguna Pueblo to be sentenced in this case. Co-defendants Preston Chino, 22, and Cameron Joseph Kasero, 21, were each sentenced last week to 96 months in federal prison followed by three years of supervised release.
Lucero, Chino and Kasero, together with co-defendant Andrea Carrillo, 22, were indicted on assault and firearms offenses in July 2013. The indictment alleged that the quartet assaulted two men and a woman on Dec. 9, 2012, by discharging firearms at a residence located in Acoma Pueblo in Cibola County, N.M.
Lucero entered a guilty plea to an assault charge on Sept. 9, 2014, and admitted repeatedly discharging a weapon at the victims’ residence.
Chino pled guilty on Aug. 27, 2014, to an assault charge, and admitted aiding Kasero and Lucero in assaulting the three victims by providing them with shotguns and ammunition. He also admitted driving his co-defendants to the victims’ Acoma Pueblo home where they discharged the shotguns multiple times in the direction of the residence.
Kasero pled guilty to an assault charge on Sept. 2, 2014. During his plea hearing, Kasero admitted assaulting the victims by discharging a shotgun at a residence while it was occupied by the victims.
Carrillo entered a guilty plea to an assault charge on Sept. 2, 2014, and admitted aiding the assault on the victims by providing her co-defendants with directions to the residence with the understanding that her co-defendants intended to commit an assault at that location. Carrillo is scheduled for sentencing on Jan. 20, 2015, when she faces a statutory maximum penalty of ten years in prison.
This case was investigated by the Albuquerque office of the FBI, the Laguna/Acoma Agency of the BIA’s Office of Justice Services, the Acoma Pueblo Tribal Police and the Pueblo of Laguna Tribal Police Department. Assistant U.S. Attorney Kyle T. Nayback is prosecuting the case.


text:
KELOWNA, BC — RCMP are investigating three Kelowna men allegedly involved in separate child exploitation offences.
The B.C. Integrated Child Exploitation (ICE) unit, which is based in Surrey, specializes in investigating internet facilitated offences against children.
Officers came to the Central Okanagan on June 24th with search warrants after undercover investigators identified the suspects.
The men are accused of offences ranging from manufacturing child pornography to the luring of a child.
Two of the men were arrested but now have been released with conditions, including restricting their future contact with children and access to internet.
The third man is under investigation and a number of his electronic devices are seized.
Investigators may recommend more charges after they finish looking through the devices.
The children in some of these Kelowna investigations have been identified as “at risk.”
One of the suspects was living with children at the time the alleged offences took place.
But BC ICE says officers do not have evidence that suggests any local children were directly victimized.
The unit uses online undercover officers to identify and arrest those who target children on the internet for sexual purpose.
“We continue to use various tools in our disposal to track down these online predators. We aim to hunt those who prey on children by using every police technique available,” adds Sergeant Van Laer.
The BC ICE unit also relies on tips from the public and encourages anyone who has information about children exploitation online to contact local police or report it at http://www.cybertip.ca.