text:
|Publication number
|Publication type
|Application number
|Publication date
|Jun 29, 2010
|Priority date
|Mar 23, 2004
|Fee status
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|Publication number
|Zeev Rosenzweig, Yongfen Chen, Desheng Wang
|Original Assignee
|University Of New Orleans Research And Technology Foundation, Inc.
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|Patent Citations (18), Non-Patent Citations (5), Referenced by (9), Classifications (35), Legal Events (4)
|External Links: USPTO, USPTO Assignment, Espacenet
Priority of our U.S. Provisional Patent Application Ser. No. 60/555,629, filed 23 Mar. 2004, incorporated herein by reference, is hereby claimed.
This work was supported by the National Science Foundation (NSF) grants CAREER CHE-9874498; NSF/LEQSF2001-04-RII-03; and CHE-0134027.
Not applicable
1. Field of the Invention
The present invention relates to markers. More particularly, the present invention relates to luminescent markers.
2. General Background of the Invention
Luminescent semiconductor quantum dots (QDs) have been intensely studied due to their unique optical properties, including sharp emission spectra with full width at half-maximum (fwhm) as narrow as 25 nm, high photostability, and tunable size dependent emission peaks.1-5 Incorporation of luminescent semiconductor quantum dots into nanospheres has been explored as a way to prepare bright biological labels and functional composite luminescent materials. For example, Barbera-Guillem successfully encapsulated luminescent CdSe QDs and other metal oxide nanocrystals in nanometric liposomes for potential biological labeling applications.6 Rogach et al. encapsulated CdSe QDs in 40-80 nm silica nanospheres and proposed to use them as building blocks to form 3D colloid crystal microstructures.7 Moffitt et al. encapsulated CdS QDs in water-soluble block copolymers to form large compound micelles (LCM) that averaged 65 nm in diameter.8
Electrostatic interactions of polyelectrolytes have been widely applied in layer-by-layer procedures to prepare thin films since Decher et al. introduced this unique technique in the early 1990s.10-11 Using this technique, Anai et al. successfully incorporated the highly charged protein avidin into a thin film and used it to immobilize biotinylated molecules.12 Caruso et al. immobilized enzymes on the surface of polystyrene microspheres to fabricate micrometric biocatalyst carriers using the same layer-by-layer approach.13 Goldman et al. prepared bioinorganic conjugates by using negatively charged CdSe—ZnS QDs and positively charged avidin. They used these new particles for luminescence immunoassay applications.14 In their study, the concentrations of QDs and avidin were limited to minimize aggregation due to electrostatic attractions.
The following U.S. Patents are incorporated herein by reference:
The following articles (and all references mentioned herein) are incorporated herein by reference:
The present invention includes a novel method to assemble highly luminescent semiconductor quantum dots (preferably CdSe—ZnS) and magnetic nanoparticles (preferably of iron oxide) into nanoassemblies. The inventors believe that this is the first time that someone has integrated luminescent semiconductor QDs and magnetic nanoparticles in the same nanoassemblies. Existing nanoparticles used in diagnostic applications of which the inventors are aware are either magnetic or luminescent but not both.
The present invention includes a novel technique to prepare luminescent and magnetic nanoassemblies averaging around 200 nm in diameter. The nanoassemblies contain hydrophilic or hydrophobic semiconductor (preferably CdSe—ZnS) quantum dots and magnetic nanoparticles of preferably iron oxide.
Hydrophilic quantum dots and magnetic particles can be encapsulated in glyconanospheres. The underlying principle of this technique is electrostatic interactions between positively charged polylysine molecules and negatively charged particles and carboxymethyldextran molecules. In addition to electrostatic attraction between the particles and the molecules, covalent amide bonds were introduced to cross-link the particles with the polysaccharide matrix to further stabilize the novel nanoassemblies.
Hydrophobic particles can be assembled in silica nanospheres using a microemulsion synthetic method.
The emission properties of the quantum dots-containing nanoassemblies were similar to the emission properties of individual quantum dots despite of the presence of magnetic nanoparticles in the assemblies, although some decrease in emission quantum yield was noted. Assembling hundreds of quantum dots in single nanoassemblies created bright and photostable nanoassemblies that could be easily observed using conventional fluorescence microscopy instrumentation. The encapsulation of magnetic nanoparticles in the nanoassemblies enabled their manipulation and separation from solution using an external magnetic field. The novel nanoassemblies could be used in biological applications requiring magnetic separation and highly sensitive fluorescence detection.
U.S. Pat. No. 6,548,171 discloses polymer microspheres. The particles of the present invention are nanometric, meaning 10-100 times smaller.
The method to load nanocrystals into the microspheres of U.S. Pat. No. 6,548,171 is by swelling and shrinking by changing solvents. The microspheres swell in one solvent which enable nanocrystals to diffuse in, then shrink in water to lock the nanocrystals in. The main problem with this approach is stability. Nanocrystals leak out of the particles when exposed to uncontrolled environments of temperature and acidity.
The method to entrap nanocrystals in the smaller particles of the present invention is different. When trapped in the glyconanospheres the nanocrystals are assembled into covalently bonded structures. They are prepared in one step, unlike the microspheres that are first prepared then loaded with nanocrystals. When trapped in silica nanospheres the nanocrystals are precursors in the microemulsion-based method. Again, the nanocrystals are covalently attached to the formed assemblies. As a result of the covalent attachment the particles of the present invention exhibit higher stability than the microspheres in which the nanocrystals are only encapsulated in through weaker physical interactions.
Since the method used to prepare the polymer microspheres in U.S. Pat. No. 6,548,171 involves swelling and shrinking it appears that the method works only with polymer materials that could swell and shrink readily when placed in different solvents. The glyconanospheres and the silica spheres of the present invention do not swell and shrink in different solvents.
U.S. Pat. No. 6,548,171 mentions that the fluorescence microspheres could have a magnetic core. There are commercially available magnetic polystyrene microspheres that could be loaded with fluorescent nanocrystals to form magnetic and luminescent microspheres. Again, these particles will be micrometric in dimensions and will take several steps to prepare. In contrast, the particles of the present invention are prepared in one step.
The preferred luminescent quantum dots of the present invention are CdSe/ZnS luminescent quantum dots: cadmium selenide nanoparticles that are coated with a zinc sulphide shell. Luminescent quantum dots that have visible emission almost always have cadmium. The counter ion could be different, for example cadmium sulphide (CdS) or cadmium teleride (CdTe). Changing the counter ion changes the emission color. CdS particles emit light at higher energies (still visible but close to the UV range). CdTe nanoparticles emit red light (still visible but close to the infrared region). In almost all cases a shell of ZnS is used to passivate the surface to increase the emission efficiency.
The preferred magnetic nanoparticles of the present invention are iron oxide magnetic nanoparticles (e.g., γ-Fe2O3 or Fe3O4). It is the understanding of the inventors that iron oxide is the only type of magnetic nanoparticles approved for in-vivo biomedical applications because these particles if capped correctly are not toxic. It is possible to form magnetic nanoparticles with superior magnetic properties like metallic cobalt or alloys of iron and gold. The iron-gold alloys could be potentially useful for cell separation applications because they may not be toxic. Cobalt particles are highly toxic (they probably should be banned for environmental concerns).
The glyconanospheres of the present invention can contain dextran molecules (or could contain other polysaccharides) on their surface that interact strongly with carbohydrate binding proteins. These glyconanospheres can be used for real-time monitoring of protein carbohydrate interactions. The inventors in their studies of the present invention focused on lectins, which are sugar binding proteins. In particular the inventors worked with Concanavaline A (Con A) which binds glycosylic residues in carbohydrates with high affinity (the dissociation constant of dextran-Con A was found in studies of the present inventors to be about 1 nanomolar).
The assay of the present invention could be carried out to screen lectins for their binding affinity to glycosylic residues on the surface of the quantum dots-containing glyconanospheres. The rate of aggregation of the glyconanospheres depends on the concentration and the binding affinity of lectins to the glycosylic residues on the surface of the glyconanospheres.
The glyconanospheres (around 200 nm in diameter) of the present invention can advantageously be used for lectin recognition assays. The quantum dots coated magnetic nanoparticles (around 30 nm in diameter) of the present invention can advantageously be used for breast cancer cells separation and detection in blood.
The present invention includes luminescent and magnetic nanoassemblies comprising luminescent semiconductor QDs and magnetic nanoparticles. These nanoassemblies can typically be around 10 nm to 300 nm in diameter. The present invention includes a plurality of the such nanoassemblies, averaging around 200 nm�25 nm in diameter or smaller. These nanoassemblies can include luminescent quantum dots and magnetic nanocrystals assembled in glyconanospheres.
The present invention includes a plurality of these nanoassemblies, which can average around 30 nm�15% in diameter or smaller. These can include nanoassemblies in which luminescent quantum dots of around 1-5 nm in diameter directly coat core magnetic nanoparticles of around 10-20 nm in diameter.
The nanoassemblies of the present invention can average around 20 nm�15% in diameter or smaller.
The nanoassemblies of the present invention can average around 20 nm�15% in diameter.
The nanoassemblies of the present invention preferably comprise luminescent semiconductor Cd-containing QDs and magnetic nanoparticles.
In the nanoassemblies of the present invention, the QDs preferably include a shell of ZnS.
In the nanoassemblies of the present invention, preferably the magnetic nanoparticles are magnetic iron oxide nanoparticles.
The nanoassemblies of the present invention preferably comprise luminescent semiconductor CdSe—ZnS QDs and magnetic iron oxide nanoparticles.
In the nanoassemblies of the present invention, preferably the nanoassemblies are encapsulated in glyconanospheres.
Preferably, the components of the nanoassemblies of the present invention are bound electrostatically. Alternatively, the components of the nanoassemblies of the present invention are bound covalently.
Preferably, the nanoassemblies of the present invention contain molecules on their surface that interact strongly with carbohydrate binding proteins. Preferably, the molecules are dextran molecules.
The nanoassemblies of the present invention preferably comprise QDs bound to the surface of thiol-modified magnetic nanoparticles.
The present invention includes a method of using the nanoassemblies of the present invention for real-time monitoring of protein-carbohydrate interactions, comprising: attaching to the nanoassemblies a protein or carbohydrate to create modified nanoassemblies; exposing the modified nanoassemblies to fluid thought to contain a carbohydrate or protein of interest to allow protein-carbohydrate interactions to occur if the fluid contains the carbohydrate or protein of interest; observing the modified nanoassemblies after the modified nanoassemblies are exposed to the fluid. Preferably, the modified nanoassemblies are separated from the fluid using magnetic force.
The present invention includes a method of using the nanoassemblies of the present invention for detecting cancer cells, comprising: attaching to the nanoassemblies antibodies against membranal biomarkers that are selectively expressed in breast cancer cells to create modified nanoassemblies; exposing the modified nanoassemblies to fluid thought to contain circulating breast cancer cells to allow the modified nanoassemblies to attach selectively to any breast cancer cells through binding of the antibody-modified nanoassemblies to membranal biomarkers on the cancer cells. Preferably, the modified nanoassemblies are separated from the fluid using magnetic force. When the fluid includes blood which contains breast cancer cells, the breast cancer cells bound to the modified nanoassemblies are separated from the blood using magnetic force. Optionally, the modified nanoassemblies can have different emission colors and different antibodies where the emission color would code for a specific antibody, to enable determination of multiple biomarkers on a cell surface.
The present invention includes a method of assembling highly luminescent semiconductor QDs and magnetic nanoparticles into nanoassemblies, comprising: providing highly luminescent semiconductor QDs in chloroform; providing magnetic nanoparticles in water; coupling the QDs to the magnetic nanoparticles in a chloroform/methanol/water mixture. Preferably, the methanol is added to the QD-containing chloroform to form a mixture of methanol and QD-containing chloroform; and the water containing magnetic nanoparticles is injected into the mixture of methanol and QD-containing chloroform under sonication and vigorous stirring. Preferably, the QDs comprise highly luminescent semiconductor CdSe—ZnS and the magnetic nanoparticles comprise iron oxide nanoparticles.
For a further understanding of the nature, objects, and advantages of the present invention, reference should be had to the following detailed description, read in conjunction with the following drawings, wherein:
The present inventors in their laboratory developed CdSe QDs doped luminescent micelles that were stabilized by a thin silica layer on their surfaces. The stabilized micelles were more stable than ordinary micelles or liposomes.9 Common disadvantages of these structures have been their lack of surface functional groups that are suitable for further bioconjugation and limited chemical stability and biocompatibility. Here, the present inventors describe the development of CdSe—ZnS QD incorporated luminescent glyconanospheres that are suitable for biomolecule conjugation while maintaining the high emission quantum yield of the QDs. The glyconanospheres preferably contain dextran molecules on their surface that interact strongly with carbohydrate binding proteins (CBP). These novel glyconanospheres could be used for real-time monitoring of protein carbohydrate interactions.
In the study in which the present invention was developed the present inventors used electrostatic interactions to incorporate negatively charged CdSe—ZnS QDs into luminescent nanospheres that contain polysaccharides on their surface. While DNA and protein molecules were already conjugated to luminescent semiconductor quantum dots,15-16 the present inventors demonstrate the synthesis of QD-polysaccharide nanocomposites. To prepare dextran grafted luminescent nanospheres, negatively charged CM-dextran and CdSe—ZnS QDs capped with mercaptosuccinic acid were mixed with 80 000 Da positively charged polylysine.
In another study in which the present invention was developed, the present inventors used electrostatic interactions to incorporate negatively charged CdSe—ZnS QDs and iron oxide magnetic nanocrystals into luminescent nanospheres that contain polysaccharides on their surface. While DNA and protein molecules were already conjugated to luminescent semiconductor quantum dots, the present inventors demonstrate for the first time the synthesis of polysaccharide nanocomposites that contain luminescent quantum dots, magnetic nanocrystals and both luminescent quantum dots and magnetic nanocrystals. To prepare dextran grafted luminescent glyconanospheres, negatively charged CM-dextran, CdSe—ZnS QDs capped with mercaptosuccinic acid and carboxyl-modified iron oxide nanocrystals were mixed with 80 000 Da positively charged polylysine.
Carboxyl-modified iron oxide nanoparticles were obtained from Indicia Biotech, France.
Carboxymethyldextran (CM-dextran) was prepared by mixing 100 mg dextran with a molecular weight of 10 000 Da with 200 mg chloroacetic acid in a 6N NaOH alkaline solution at 60� C. for 1 h following a procedure first suggested by Rebizak et al.18 To prepare the luminescent glyconanospheres, 300 μL aqueous solution of 500 μM CM-dextran was mixed with 120 μL aqueous solution of 0.19 μL mercaptosuccinic acid modified CdSe—ZnS QDs (when forming QDs containing spheres), 0.19 μL of carboxyl-modified iron oxide nanoparticles (when forming iron oxide containing spheres) and 0.19 μL of CdSe/ZnS QDs and 0.19 μL iron oxide nanoparticles (when forming luminescent/magnetic nanocomposite spheres). The concentration of the CdSe—ZnS QDs was calculated using molecular extinction coefficients previously reported by Striolo et al. of 1.1�106 M−1 cm−1 for 3 nm sized CdSe—ZnS QDs and 1.22�106 M−1 cm−1 for 6 nm sized CdSe—ZnS QDs.19 Then, 120 μl of 10 μM polylysine in a pH 7.0 phosphate buffer solution was added to the CM-dextran and QD containing solution. The reaction mixture was incubated at room temperature for 30 min. The concentration of iron oxide nanoparticles was calculated based on inductive coupled plasma (ICP) atomic absorption measurements that quantified the level of iron per mg sample. The newly formed glyconanospheres were washed three times using centrifugation at 4500 rpm for 15 min to remove unreacted QDs and CM-dextran.
Since this preparation method is based on electrostatic interactions, the size of the QDs should not affect the formation of the nanospheres (small variations in the size of the quantum dots or magnetic nanocrystals did not affect the formation of the larger glyconanospheres). Indeed, QDs of different diameter and different emission color were incorporated into the luminescent glyconanospheres.
The dextran molecules play an important role in forming the spherical nanospheres and preventing their aggregation.
To prove that dextran molecules are indeed incorporated into the nanospheres, the present inventors prepared glyconanospheres with Oregon Green labeled dextran molecules. Oregon-Green is a highly luminescent green dye with fluorescein-like emission properties (λex=470 nm, λem=525 nm). It also provides one negative charge for each dye molecule that is attached to the dextran polymeric chain. When excited at 470 nm, the glyconanospheres showed bright green emission due to the emission of Oregon green (
Unlike the strong attraction of polyelectrolytes often observed in thin films formed on a flat surface using the layer-by-layer deposition technique, the present inventors found that electro-static interactions solely are not strong enough to stabilize the luminescent glyconanospheres. Fluorescence microscopy measurements revealed that these luminescent glyconanospheres dissociated in about 10 h when stored in aqueous solution at room temperature. The relative instability of the electrostatically held glyconanospheres could be attributed to their large surface-to-volume ratio. The interactions of solution ions with surface charges weaken the electrostatic attraction between the positively charged polylysine and the negatively charged CM-dextran, and the mercaptosuccinic acid modified CdSe QDs. To further stabilize the luminescent glyconanospheres, the present inventors introduced the standard water-soluble bioconjugate coupling agent 1-ethyl-3-(3)-dimethylamino-propyl carbodiimide (EDC) to initiate the formation of covalent bonds between the carboxylic groups on the surface of the mercaptosuccinic acid modified CdSe QDs and in the CM-dextran chains and the amino groups of the polylysine chains. The concentration of EDC was 500 mM. The reaction mixture was incubated at room temperature for 2 hours. The formation of amide bonds between the carboxylic and amino groups in the glyconanospheres greatly increased their chemical stability. Luminescence microscopy measurements revealed that the covalently and electrostatically held glyconanospheres remained stable for over two months when stored in aqueous solution at room temperature.
CM-dextran provides the glyconanospheres with high surface density of glucosylic residues. Thus, the carbohydrate binding protein concanavalin A (Con A) can recognize the surface bound dextran. Con A is a lectinic protein that selectively binds to the terminal residues of R-D-glucose and R-D-mannose. Each Con A molecule contains four binding sites for carbohydrates.20 These polyvalent binding sites have been often used in cell agglutination and separation of glycoproteins in affinity chromatography.21 To demonstrate the binding interactions of the surface-bound dextran and Con A, the present inventors mixed a solution of 200 μL 0.04 μM (based on the concentration of CdSe—ZnS QDs) glyconanospheres with 2 mL 0.05M HEPES buffer solution (pH 7.2) containing 0.25 mg/mL Con A, 0.1 mM Mn2+, and 0.1 mM Ca2+. The solution was incubated at room temperature for 2 h.
The present inventors have developed a novel method to assemble highly luminescent semiconductor CdSe—ZnS QDs into glyconanospheres through electrostatic interactions and covalent stabilization. The present inventors have also developed a novel method to assemble highly luminescent semiconductor CdSe—ZnS QDs and iron oxide nanocrystals into glyconanospheres through electrostatic interactions and covalent stabilization. The emission properties of the QDs containing glyconanospheres were similar to the emission properties of individual QDs. Assembling hundreds of QDs in single glyconanospheres created bright and photostable particles that could be easily observed using conventional fluorescence microscopy instrumentation. This could make luminescent QDs more accessible to fluorescence microscopy studies of biological samples. For example, the luminescent glyconanospheres showed high affinity to Con A, a glucose specific lectin. As demonstrated herein, these glyconanospheres could be used to study carbohydrate-protein interactions.
Superparamagnetic Fe2O3 Beads-CdSe/ZnS Quantum Dots Core-Shell Nanocomposite Particles for Cell Separation
A preferred embodiment of the present invention includes the formation of luminescent and magnetic nanocomposite particles that consist of superparamagnetic core particles (γ-Fe2O3) and a layer of luminescent CdSe/ZnS quantum dots (QDs) on their surface. These particles could be used in a variety of biomedical and biological applications including magnetic separation and detection of cancer cells, bacteria and viruses. CdSe semiconductor quantum dots (QDs) with diameters ranging between 1.5 and 8 nm exhibit strong, tunable luminescence.22-26 They have been widely investigated for their size-dependent optoelectronic properties,27 and for their potential use in optical devices,28 biological labels,29 and sensors.30 Luminescent QDs are brighter, exhibit higher photostability and show narrower emission peaks compared to organic fluorophores.29 CdSe QDs are often coated with a layer of ZnS, which has a larger band gap, to passivate their surface and to increase their emission quantum yield through quantum confinement.31 To enable miscibility of the QDs in aqueous solution, their surface has been modified with carboxyl functional groups.32 Alternatively, QDs were encapsulated in water miscible shells such as liposomes,33 silica miscelles,34 and glyconanospheres.35 QDs were also encapsulated in dendrimers36,37 to increase their stability against photooxidation and photobleaching.
Luminescent CdSe/ZnS QDs of green emission and red emission colors were synthesized based on a method developed by Peng et al., with minor modifications.25 12.7 mg cadmium oxide and 160 mg lauric acid were mixed in a 100 mL three-neck flask. The mixture was heated to ˜200� C. in a mantle to fully dissolve the cadmium oxide in the lauric acid solution. Then, 1.94 g trioctylphosphine oxide (TOPO) and 1.94 g hexadecylamine (HDA) were added to the solution. The mixture was heated to a temperature higher than 280� C. and kept under a dry nitrogen atmosphere. Upon reaching the desirable temperature the mantle was removed and 2 mL trioctylphosphine (TOP) solution containing 80 mg selenium powder was rapidly injected into the solution under vigorous stirring. It was previously shown that the diameter of the formed QDs depends on the reaction temperature with smaller particles formed at higher temperature.22 The color of the mixture changed from clear colorless to yellow, orange, or red depending on the exact temperature. To form a ZnS coating on the CdSe QDs, the mixture was cooled to ˜200� C. Then, after being kept three minutes at this temperature, a solution containing 250 μl hexamethyldisllathiane ((TMS)2S) and 1 mL diethylzinc (Zn(Et)2) premixed in 2 mL TOP was gradually injected into the solution over a minute. The reaction mixture was kept at 180� C. and stirred for 1 h. The solution was cooled to room temperature and the resulting sample of CdSe/ZnS QDs was washed three times with methanol and chloroform. The QDs showed extremely high quantum yield, reaching 100%. The present inventors even can see the shining fluorescence under room light.
Polymer coated γ-Fe2O3 superparamagnetic magnetic beads were purchased from Indicia Biotech, France. The nanometric γ-Fe2O3 magnetic particles were coated with dimercapto-succinimid acid (DMSA) to stabilize and functionalize the ferrofluid. The surface of the ferrofluid was covered with free thiol (SH) and carboxyl (COOH) residues (3:20 thiol to COOH ratio) to enable covalent coupling of various ligands to the magnetic particles.38 A TEM image (
The coupling between the luminescent CdSe/ZnS QDs and the magnetic beads was based on thiol chemistry. Thiols (—SH) are probably the most utilized functional groups for stabilizing and modifying CdSe QDs.32,36,37,39,40 Thiol groups form stable bonds with metals on the surface of QDs such as cadmium and zinc.41-45 However, the coupling reaction between the QDs and magnetic beads presented a difficulty since the trioctylphosphine oxide (TOPO) capped CdSe/ZnS QDs were dissolved in chloroform while the polymer-coated magnetic beads were dispersed in water. The present inventors found that running the coupling reaction in a 10:5:1 mixture of chloroform/methanol/water yielded the best nanocomposite particles with minimal aggregation. To carry out the coupling reaction the present inventors first transferred 1 mL of 1 μM CdSe/ZnS QDs into a 5 mL vial. Then the present inventors added 500 μl methanol to the solution. This was followed by the slow injection of 100 μl 0.1 μM magnetic bead-water suspension to the solution under sonication and vigorous stirring. Under these reaction conditions the molar ratio between the QDs and the mage ic particles was 100:1. The excess of QDs was imperative to preventing aggregation of the magnetic beads. Under vigorous stirring the aqueous and organic phases formed an even suspension. The suspension was stirred for 1 h to form the nanocomposite particles. The luminescent/magnetic nanoparticles were then separated from the solution by using a permanent magnet (average cross sectional force density 16.6 T2/m) and washed several times with methanol. The sample was vortexed briefly and sonicated for 15 min to prevent aggregation.
A representative TEM image of the nanocomposite particles is shown in
N=2π(R Fe +R QD)2/✓3R QD 2
where RFe is the radius of the magnetic beads and RQD is the radius of the smaller QDs. This estimated expression is derived by dividing the surface area covered by the small QDs on the larger iron oxide particle by the area covered by a single QD on the iron oxide particle surface. The calculation assumes close packing of QDs on the magnetic particle surface and takes into consideration the gaps between the QDs. For example, for a magnetic particle diameter of 10 nm, the maximum number of QDs increases from 45 to 133 when the QD diameter decreases from 4 to 2 nm. A high-resolution TEM image of an individual magnetic bead coated with CdSe/ZnS QDs is shown in
Luminescence images of the nanocomposite particles coated with ˜3 and ˜5 nm CdSe/ZnS QDs are shown in
To apply the luminescent/magnetic particles for magnetic separation of cells followed by luminescence detection, the present inventors first modified their surface with carboxylic functional groups. A 2 mL portion of 0.1 μM luminescent/magnetic nanoparticle suspension in methanol was transferred into a 250 mL flask, which was heated to 60� C. under stirring under reflux for 6 h. Then, 200 μl mercaptoacetic acid (Sigma) was injected into the flask. The reaction took 1 h to complete. The carboxyl-modified nanoparticles were washed three times using magnetic decantation and were finally dispersed in 2 mL deionized water and briefly vortexed to prevent aggregation. As expected, the carboxy-modified nanoparticles were highly water-soluble and also maintained their luminescence properties.32,40 Mouse anticycline E antibody molecules were attached to the particles through EDAC (1-ethyl-3-(3-dimethylaminopropyl) carbodiimide) coupling as shown in
A 10 μl portion of mouse anticycline E antibody (Zymed) solution was added into 1 ml ˜0.1 μM luminescent/magnetic nanoparticles suspension in a phosphate buffer (PB) solution at pH 7.4. Then 100 mg EDAC coupling reagent was added into the mixture. The mixture was gently shaken for 1 h and the antibody-labeled particles were then separated and washed using magnetic decantation. The anticycline E labeled nanoparticles were resuspended in 1 mL PB solution at pH 7.4. The use of EDAC coupling enabled the covalent attachment of the antibodies to the carboxyl-modified particles without substantially damaging the active site of the antibodies. To demonstrate the separation capability of the particles the present inventors incubated a sample of 200 μl anticycline E labeled luminescent/magnetic particles with a 1 mL MCF-7 breast cancer cell suspension containing 10,000 cells/mL. Following 15 min incubation at room temperature under gentle shaking the cells were separated from the suspension using a permanent magnet. The cells were washed twice with a phosphate buffer solution using magnetic decantation and observed using digital fluorescence imaging microscopy. Transmission and fluorescence images of the MCF-7 cells labeled with the anticycline E labeled particles are shown in
Conclusions. Water-soluble nanocomposite particles consisting of a magnetic core (γ-Fe2O3) and luminescent quantum dots shell (CdSe/ZnS QDs) were synthesized in an organic/water two-phase mixture. Thiol chemistry was used to bind the QDs to the surface of the magnetic beads. The luminescent/magnetic nanocomposite particles were characterized using TEM and EDS analysis. The particles averaged 20 nm in diameter with about 15% size variation, showed relatively smooth morphology, and were fully water miscible. They also exhibited high emission quantum yield and were easily separated from solution using a permanent magnet. Anticycline E molecules against the breast cancer specific marker cycline E, were attached to the luminescent/magnetic particles followed their functionalization with carboxylic groups through EDAC coupling. The anti cycline E labeled particles were used successfully to separate and detect breast cancer cells in serum. The separation capabilities of the antibody labeled particles in cell suspensions containing MCF-7 and red blood cells are currently under investigation.
Incorporated herein by reference is "Synthesis of Glyconanospheres Containing Luminescent CdSe—ZnS Quantum Dots" by Yongfen Chen, Tianhao Ji, and Zeev Rosenzweig, published in Nano Lett., Vol. 3, No. 5, 2003, 581-584 (copy attached to U.S. Provisional Patent Application Ser. No. 60/555,629, filed 23 Mar. 2004).
Also incorporated herein by reference is "Superparamagnetic Fe2O3 Beads-CdSe/ZnS Quantum Dots Core-Shell Nanocomposite Particles for Cell Separation" by Desheng Wang, Jibao He, Nista Rosenzweig, and Zeev Rosenzweig, published in Nano Lett., 2004, Vol. 4, No. 3, 409-413 (copy attached to U.S. Provisional Patent Application Ser. No. 60/555,629, filed 23 Mar. 2004).
All measurements disclosed herein are at standard temperature and pressure, at sea level on Earth, unless indicated otherwise. All materials used or intended to be used in a human being are biocompatible, unless indicated otherwise.
The foregoing embodiments are presented by way of example only; the scope of the present invention is to be limited only by the following claims.
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|Apr 5, 2012
|Oct 1, 2013
|Sony Corporation
|Microbead automatic recognition method and microbead
|Feb 17, 2012
|Apr 7, 2015
|Senior Scientific Llc
|Magnetic needle biopsy
|Nov 5, 2010
|Aug 4, 2015
|Senior Scientific Llc
|Detection, measurement, and imaging of cells such as cancer and other biologic substances using targeted nanoparticles and magnetic properties thereof
|Apr 9, 2009
|Nov 12, 2009
|Agency For Science, Technology And Research
|Coated composites of magnetic material and quantum dots
|May 16, 2006
|Dec 3, 2009
|Innova S.A.
|Sweetener compositions
|Sep 26, 2008
|Nov 11, 2010
|Nanoco Technologies Limited
|Nanoparticles and their manufacture
|Jan 20, 2011
|National Chiao Tung University
|Drug Delivery Nanodevice, its Preparation Method and Uses Thereof
|Aug 2, 2012
|Sony Corporation
|Microbead automatic recognition method and microbead
|U.S. Classification
|428/402, 977/783, 428/407, 436/526, 435/103, 977/838, 977/773, 977/830, 977/834, 428/403, 977/824
|International Classification
|Cooperative Classification
|European Classification
|Jul 6, 2005
Owner name: UNIVERSITY OF NEW ORLEANS RESEARCH AND TECHNOLOGY
Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:ROSENZWEIG, ZEEV;CHEN, YONGFEN;WANG, DESHENG;SIGNING DATES FROM 20050619 TO 20050621;REEL/FRAME:016227/0718
|Feb 7, 2014
|Maintenance fee reminder mailed
|Jun 29, 2014
|Lapse for failure to pay maintenance fees
|Aug 19, 2014
|Expired due to failure to pay maintenance fee
Effective date: 20140629


text: Rebel Wilson By Maggie Malach April 12, 2015 6:03 PM | PopCrush Share on Facebook Share on Twitter Rebel Wilson at the 2015 MTV Movie Awards. Source: Rebel Wilson Category: Share on Facebook Share on Twitter More Leave a comment Send to a friend! Print this page Pin it! Reddit This! Share on Tumblr! Best of KXLB Bozeman Has a New Downtown Bakery Bozeman Area Fourth of July Events Summer Solstice, Full Moon and Bozeman Longest Day Event Happening Today Laney Lou and the Bird Dogs Perform at Headwaters Country Jam Recommended For You ZergNet Best of the Web ZergNet

subset: commoncrawl


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Please remember to follow the manual of style and code of conduct at all times.
Check BNN and Bulbanews for up-to-date Pokémon news and discuss it on the forums or in our IRC channel #bulbagarden on irc.systemnet.info.
Check BNN and Bulbanews for up-to-date Pokémon news and discuss it on the forums or in our IRC channel #bulbagarden on irc.systemnet.info.
Cacturne (Plasma Freeze 10)
From Bulbapedia, the community-driven Pokémon encyclopedia.
Cacturne (Japanese: ノクタス Noctus) is a Grass-type Stage 1 Pokémon card. It is part of the Plasma Freeze expansion.
Card text
Pokédex data
|Cacturne - Scarecrow Pokémon
|4'03" (1.3 m)
|170.6 lbs. (77.4 kg)
|Pokédex entry
|It becomes active at night, seeking prey that is exhausted from the day's desert heat.
|よるになると かつどうを はじめる。さばくの あつさに つかれはてた えものを みつけだし とらえるのだ。
Payback is a move in the Pokémon games that Cacturne can learn. This card's Pokédex entry comes from Pokémon Black 2 and White 2.
|This article is part of Project TCG, a Bulbapedia project that aims to report on every aspect of the Pokémon Trading Card Game.
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The Immigrant Left in the United Statesby Paul Buhle
View All Available Formats & Editions
This book investigates the role immigrant radicals have played in U.S. society from the mid-nineteenth century to the present.
Editorial Reviews
"I like very much that this book covers immigrant life and politics over the long run of U.S. history rather than focusing on either the past or the present. Essays on immigrant workers inevitably shed light on nationalism, transnationalism, internationalism, and their significance for class analysis. The foreign-language labor and Left press has been surveyed, and evaluated, but this rich source has not yet been mined extensively." -- Donna Gabaccia, University of North Carolina at Charlotte
Product Details
- 9780791428832
- Publisher:
- State University of New York Press
- Publication date:
- 04/28/1996
- Series:
- SUNY series in American Labor History Series
- Pages:
- 349
Meet the Author
Paul Buhle directs the Oral History of the American Left project, Tamiment Library, New York University, and has written or edited 21 books, including Marxism in the United States; Encyclopedia of the American Left; The American Radical; and C. L. R. James: The Artist as Revolutionary.
Dan Georgakas teaches in the Labor Education and Advancement Project of Queens College. He is author of Greek America at Work; associate editor of the newspaper The GreekAmerican; and coeditor of Encyclopedia of the American Left and New Directions in Greek American Studies.
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→ TIP: To search for an exact phrase, put quotes around it. For example, "example of a prayer".
Prayers for Today G-MANS JOURNEY - June 26, 2016 ... (Prayer of Ignatius Loyola, 1491-1556 ) Lord, I wish to put aside all distractions and to give you my total focus. I will do nothing more important today than to meditate prayerfully on your goodness and your active role in my life. Though I am unworthy to be in your presence, I trust in your mercy and love. Through this moment of prayer I want to draw closer to you and learn to live more like ...http://gman59.blogspot.com/2016/06/prayers-for-today.html
Sleep TIGERISH WATERS - June 26, 2016 The efficacy and power of prayer does not come from us. Prayer comes through us but it can leave us drained. In order for Joshua to be successful in battle, Moses has to pray and if he got tired and his arms dropped things did not go well for Joshua. So Aaron and Hur supported his arms. The analogy to the work of the Church and Christ's work is blatantly obvious. Yes, prayer drains. The woman with the issue of blood drained the strength ...http://tigerishwaters.blogspot.com/2016/06/sleep.html
Prayer: Paul VI IGNATIAN SPIRITUALITY: SET THE WORLD ABLAZE - June 26, 2016 God of peace, we bless you and we thank you because you have sent us your beloved Son, Jesus, the source of all peace. Open yet more our hearts to the needs of all our brothers and sisters, so that we may be better able to build a true peace. Remember all who are in pain, who suffer and died in the cause of a more just world. For the people of every race, of every tongue – may your kingdom come.http://predmore.blogspot.com/2016/06/prayer-paul-vi.html
Centering Prayer and Distractions ASSOCIATION OF CATHOLIC WOMEN BLOGGERS - June 25, 2016 One Centering Prayer practitioner wrote to me in an online conversation, "In this information age of constant texting and tweets I think Centering Prayer is a very practical way to do 'be still' and surrender in God's presence; it's helped me to detach from negative thoughts and feelings…" This is a common assertion, one I'm sympathetic to. We certainly do have too much "noise" in our world–not just beeps and drum beats, but also texts and ...http://associationofcatholicwomenbloggers.blogspot.com/2016/06/centering-prayer-and-distractions.html
Tea Time With Tiffany #54 - Prayer Life Routine in the Midst of the Whirlwind... LIFE OF A CATHOLIC LIBRARIAN - June 24, 2016 ... a cathartic summer edition of: Today I talk about the whirlwind activities going on in my life right now, and how I long for the solace of a prayer routine in the midst of it. Organization, sometimes I miss you so much. *heart* ***To subscribe to the audio version of Tea Time with Tiffany, just search for it in iTunes or use this link to subscribe via Feedburner in your podcatcher of choice. Intro music is "Tea Ceremony" from PlayonLoop.com Items mentioned in this episode: My ...http://www.lifeofacatholiclibrarian.com/2016/06/tea-time-with-tiffany-54-prayer-life.html
Even More Intercession A WING AND A PRAYER - June 24, 2016 Once again, I seek stable, suitable employment. Back to this prayer again (this will be the fourth time I've used it in a post). ++++++++++ Prayer to Saint Anthony of Padua: Good Saint Anthony, in God's providence you have secured for His people many marvelous favors. You have been especially celebrated, good Saint Anthony, for your goodness to the poor and the hungry, for finding employment for those seeking it , for your special care of those who travel, ...http://awingandaprayer.blogspot.com/2016/06/even-more-intercession.html
Prayer: "Prayer for Compassion" by Teilhard De Chardin, S.J. IGNATIAN SPIRITUALITY: SET THE WORLD ABLAZE - June 23, 2016 O God, I wish from now on to be the first to become conscious of all that the world loves, pursues, and suffers; I want to be the first to seek, to sympathize, and to suffer; the first to unfold and sacrifice myself, to become more widely human and more nobly of the earth than any of the world's servants.http://predmore.blogspot.com/2016/06/prayer-prayer-for-compassion-by.html
Pause for Prayer: FRIDAY 6/24 A CONCORD PASTOR COMMENTS - June 23, 2016 Lord, what will you ask of me today? - my time? - my forgiveness of someone who's hurt me? - a change of mind? - a change of heart? - a change of habit? - outreach to someone in need? - a return to the path I belong on? - a little more honesty with myself? - some time in prayer with you? - something known only to you and me? Help me know what you ask of me today, Lord, and help me do it - today... Tweet Subscribe to A Concord Pastor Commentshttp://concordpastor.blogspot.com/2016/06/pause-for-prayer-friday-624.html
5th Sunday After Pentecost 19 Jun 2016 Sermon LUX VERA - June 23, 2016 5th Sunday after Pentecost 19.6.16 Unity in Prayer The teachings on charity, as contained in today's Gospel (Mt 5,20-24), are demanding, but are not meant to be out of our reach. Our Lord is raising the command on charity to a higher level. Now, not only do we do not kill our brother; we do not even call him a fool! Such is the level of unity and concord that Our Lord desires for us – especially for those who profess to be His disciples. And even more so again for ...http://luxvera.blogspot.com/2016/06/5th-sunday-after-pentecost-19-jun-2016.html
Solemnity Alert! COFFEE AND CANTICLES - June 23, 2016 Don't forget to do Evening Prayer I for St. John the Baptist tonight. John the Baptist by Jacek Malczewski (wikimedia commons)http://feedproxy.google.com/~r/CoffeeAndCanticles-TheDivineOfficeInYourLife/~3/ZdSXHVSLpLs/solemnity-alert.html
To Pray Is Human ASSOCIATION OF CATHOLIC WOMEN BLOGGERS - June 22, 2016 Since starting my new website Is Centering Prayer Catholic?, I've been pondering why certain errors about prayer are so common. I think I've hit on a root misunderstanding that's at the bottom of many of these errors: ignoring the distinction between the natural and the supernatural in prayer. The Catechism quotes St. John Damascene in saying: " Prayer is the raising of one's mind and heart to God or the requesting of ...http://associationofcatholicwomenbloggers.blogspot.com/2016/06/to-pray-is-human.html
'Stay With Me, Lord' by Saint Pio of Pietrelcina GOD IS AT WORK IN YOU! - June 22, 2016 A new version of this magnificent prayer by Saint Pio, for you for the sake of all, thanks be to God and His Immaculate Mother. What wonders. A prayer written by Saint Pio for the sacred fifteen minutes when one's body and soul are Divinely unified with the Real Presence of Jesus the Divine Bridegroom of every soul in the Blessed Sacrament, the greatest Source of grace we have on earth, the Compendium of all Miracles (Saint Thomas), and our Remedy for ...http://godisatworkinyou.blogspot.com/2016/06/stay-with-me-lord-by-saint-pio-of.html
Speak Lord. Your Servant Is Listening. HAPPY CATHOLIC* - June 22, 2016 Breton Girls at Prayer William-Adolphe Bouguereau, 1904 via French Painters Before the Lord the whole universe is as a grain from a balance or a drop of morning dew come down upon the earth. But you have mercy on all, because you can do all things; and you overlook people's sins that they may repent. For you love all things that are and loathe nothing that you have made; for what you hated, you would not have fashioned. And how could a thing remain, unless you willed it; or ...http://happycatholic.blogspot.com/2016/06/speak-lord-your-servant-is-listening_22.html
Revisiting the Liturgy of the Hours THE CLOISTERED HEART - June 22, 2016 ... prayer? I could not have been more wrong. The Liturgy of the Hours, also known as the 'Divine Office,' is an official group of prayers used by priests and Religious. It is a primary part of the daily schedules of monks and nuns. The Divine Office is the same for people throughout the Church, throughout the world. On the very same day, Father O'Neill in Dublin and a group of monks in Sydney and a monastery of nuns in Toledo are praying. And I can pray with them, if I wish. As I ...http://www.thecloisteredheart.org/2016/06/revisiting-liturgy-of-hours_22.html
The Energy of Christian Prayer CATHOLIC LIGHT - RADICAL FAITH & TRUST - June 21, 2016 The Energy of Christian Prayer : Christian prayer is rooted in the truth that Christ crucified is fully God and fully man. The prayer of the Lord is theandric — the highest expression of the perfect unity of divine and human energy in His Divine Person. Theandric comes from the Greek words for God (Theos) and for man (anthropos). To say that …http://tsahaylu.blogspot.com/2016/06/the-energy-of-christian-prayer.html
Re-Reading Your Week? REFLECTIONS OF AN RSCJ - June 21, 2016 http://reflectionsofanrscj.blogspot.com/2016/06/re-reading-your-week.html
Pause for Prayer: WEDNESDAY 6/22 A CONCORD PASTOR COMMENTS - June 21, 2016 Tweet Subscribe to A Concord Pastor Commentshttp://concordpastor.blogspot.com/2016/06/pause-for-prayer-wednesday-622.html
Prayer: Patience and Understanding JOURNEY OF A CATHOLIC NERD WRITER - June 21, 2016 ... and dishonest boyfriend I've ever had. Fast forward several years later. As I grew spiritually, I learned that sometimes our prayers don't get answered because it's not God's will for us; that He has something better in store. It wasn't overnight. Some of my reversion prayers included a lot of bargaining (big no-no!) so I'd get what I wanted. Eventually I began accepting that I could ask for something but that it wouldn't always be answered. I accepted it and tried to see the positive in ...http://catholicnerdwriter.blogspot.com/2016/06/prayer-patience-and-understanding.html
O My God, Trinity Whom I Adore – Blessed Elisabeth of the Trinity Prayer CATHOLIC LIGHT - RADICAL FAITH & TRUST - June 20, 2016 O My God, Trinity Whom I Adore – Blessed Elisabeth of the Trinity Prayer : O My God, Trinity Whom I Adore Blessed Elisabeth of the Trinity Prayer As we mentioned earlier this year, on March 3rd, Pope Francis approved Blessed Elisabeth of the Trinity for canonization. Today, a meeting of cardinals is expected to set the date for when she will be raised to the altars. Blessed Elisabeth of …http://tsahaylu.blogspot.com/2016/06/o-my-god-trinity-whom-i-adore-blessed.html
The Holy Father's Prayer Intentions for July 2016 BIG C CATHOLICS - June 20, 2016 Please remember the Holy Father Pope Francis' intentions in prayer through the month of July: General Intention: Indigenous Peoples That indigenous peoples, whose identity and very existence are threatened, will be shown due respect. Evangelization: Latin America and the Caribbean That the Church in Latin America and the Caribbean, by means of her mission to the continent, may announce thehttp://www.bigccatholics.com/2016/06/the-holy-fathers-prayer-intentions-for.html


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What to do with doubt I often have people email me, saying, "I want to believe, but I still have doubts. What can I do? The first thing I think we need to realize is that doubts are a common, emotional experience which need not challenge our commitment. C.... [Read more...]
What To Do With Doubt
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Global Account Manager, Downstream and Chemicals
Swift Worldwide Resources
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Armenian chess players successful in Dubai Open first round
April 16, 2012 - 16:28 AMT
PanARMENIAN.Net - The first round of Dubai Open Chess Championship kicked off on April 15.
Vladimir Akopian scored a win over Ali Elmejbri (Lebanon), Tigran L. Petrosian beat Mihail Chommyyev (Turkmenistan), Robert Hovhannisyan won John Bennett (England). Zaven Andriasian gained a victory over Zhansaya Abdumalik (Kazakhstan), Avetik Grigoryan- Maria Blanco (Venezuela), Davit Petrosian – Mama Ingcongan.
Karen Grigoryan beat Elnemr Mohamed (Egypypt). Vahe Baghdasaryan was the only Armenian representative who lost to Shanepes Meredow (Turkmenistan).
Dubai Open Chess Championship is held in 9 round Swiss system. The winner will get USD 10 thous. Contestants to gain 2-3places will be awarded USD 7 thous. and 4 thous. respectively.
Top stories
Raiola told German newspaper Bild he is happy to fight for what his client wants, which is to play under Jose Mourinho next season.
"Such an offer to join Manchester United might only come once in a lifetime for a player," Mkhitaryan's agent Mino Raiola said.
Due to the temporary disqualification, the athlete can't participate in tournaments, also set to miss the Rio Olympics in August.
"I am a positive guy. If I wasn't confident of reaching some kind of conclusion, I wouldn't try," the agent of Henrikh Mkhitaryan said.
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Iraqi army closes in on IS militants near Falluja Backed by air strikes from a U.S.-led coalition, Iraqi artillery bombarded targets as its troops closed in on up to 150 insurgents.
At least 35 killed in triple Islamic State bomb attack in Yemen An offshoot of al-Qaeda controlled the port city until recently. It was recaptured in April by Yemeni govt. and Saudi-led coalition forces.
Damaged flight data recorder from EgyptAir plane "repaired" Flight MS804 from Paris to Cairo plunged into the Mediterranean Sea on 19 May, killing all 66 people on board.


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3 colors
JS Collections Short Sleeve Soutache Cocktail Dress
Women's JS Collections Lace & Pleat Panel Crepe
Rich black crepe frames the blushing lace-covered bodice and accordion-pleat skirt to imbue this elongated A-line gown with romantic two-tone dimension. A solid inset waistband further defines the feminine silhouette. 60 1/2" length (size 8). Hidden back-zip closure. Fully lined. 95% polyester, 5% spandex. Dry clean. By JS Collections; imported. ... Learn More
Women's Cheap Monday 'Mystic' Stripe Swing Dress, Size
A trim, mock-neck bodice releases into a full, flowing skirt on a pieced swing dress wrapped in twisting, multidirectional stripes. 35" length (size Medium). Slips on over head. Mock neck. Short sleeves. Unlined. 100% organic cotton. Machine wash warm, line dry. By Cheap Monday; imported. BP. Helpful info: Dress Fit ... Learn More
Women's Sean Collection Mixed Media Fit & Flare
An incognito illusion yoke framed in stylized lace kicks of the delicate details throughout this flirty cocktail dress. Shutter-pleated satin shows off the narrow waist and the swirling hemline brings the layered tulle skirt to life. 35" length. Side zip closure. Cap-sleeve illusion yoke. Partially lined. 100% polyester. Dry clean. ... Learn More
Women's St. John Collection Embellished Degrade Croc Knit
A silvery jacquard crocodile pattern emboldens curves in a square-neck sheath knit from shimmering yarns that fade from glowing gold to glinting black. Hand-placed Austrian crystals sprinkle the hem to enhance the glamorous dégradé effect. 38 1/2" length (size 8). Hidden back-zip closure. Square neck. Elbow-length sleeves. 32% polyester, 30% ... Learn More
Women's Versace Collection Print Georgette Silk Dress, Size
Modern and Baroque design combine to graphic effect on this stretch-georgette dress printed in an artful mix of colorful stars and swirling filigree. Softly sheer flared sleeves echo the swing babydoll-style silhouette, which is loosely defined by rows of metallic square studs at the Empire waist. Hidden back-zip closure. Notch ... Learn More
Women's Pamella Roland Embellished Silk Faille Fit &
A graceful floral-and-leaf motif rendered in metallic paillettes sweeps across a shockingly pink silk-faille frock that's meticulously crafted in a fit-and-flare silhouette. A slender décolleté keyhole brings daring allure to the retro-femme design. Let our experts help perfect the fit. Learn more. 44 1/2" length (size 10). Hidden back-zip closure. ... Learn More
Women's Misha Collection 'Alessandra' Backless Silk Midi Dress,
A completely open back and soaring side slits show off some skin in this chic midi-length cocktail dress crafted from luxuriously white silk. 47" length (size Medium). Back zip closure. Square neck. Spaghetti straps. Fully lined. 100% silk. Dry clean. By Misha Collection; imported. Special Occasion. Helpful info: Dress Fit ... Learn More
Women's St. John Collection Crystal Embellished Shimmer Milano
A semi-sheer silk-georgette yoke graces an elegant gown boasting brilliant Austrian crystals that complement the subtle glint of the shimmer-infused knit. A high front slit eases movement in the column silhouette and offers a beguiling glimpse of leg. 62" length. Hidden back zip with button-and-loop keyhole closure. Jewel neck. Short ... Learn More


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One killed, 20 injured in Thane train collision
Mumbai: At least one woman was killed and another 20 passengers injured when the Vidarbha Express rammed into four derailed coaches of a Mumbai suburban train in Thane district, officials said here Thursday.
Teams of senior officials, medicos and relief workers have been rushed to the scene of the double accident between Kasara and Umbermali, said a Central Railway official.
The accident occurred around 9.30 p.m., when four coaches of the Kasara-CST suburban local train derailed and fell across the parallel railway line.
A few minutes later, the speeding 12105 Mumbai-Gondia Vidarbha Express rammed into the derailed bogies scattered on its route towards Kasara.
As a result, the engine of the express train also derailed and blocked the services on both sides.
At present, suburban services and long-distance trains are being operated only till Asangaon on the CST-Satara section and efforts were continuing to clear both the railway tracks.
Heavy rains in the Thane-Raigad belt was hampering the rescue and relief operations which are likely to continue till Friday.
The CR has set up two helpline numbers for the benefit of the victims` relatives: 020-22694040/ 10720.
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In the aftermath of their BLACKEST NIGHT encounter, Vril Dox and his team realize they must kill Starro the Conqueror. It's a job that requires three dangerous missions led by Dox, Captain Comet and Adam Strange, who will face loss, treachery and the overwhelming might of Starro's conquering horde.
"The Flash" Adds "Harry Potter" Star Tom Felton as Series Regular


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rss.feedsportal.com – The true story of a real fake filmWe've been following the development of Ben Affleck's latest directorial effort, Argo, with interest for some time now. He's plucked a great script, gathered an amazing cast, applied everything he's learned from Gone Baby Gone and The Town, and the fruits of his labour are now on screen in the first trailer for the movie.
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It took awhile for donnieyen to find his groove, but when he did, he proved he could choose headlines with the best of them.
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1. Finding DoryGross: $135,060,273Total: $135,060,273
2. Central IntelligenceGross: $35,535,250Total: $35,535,250
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Want to list your puppies for sale? Post a classified today!
About Redwing Kennel
We breed IRW & IRS and Gordon's for the field. We hunt quail and phesant and like our dogs to foot handle and retrieve. We do compete in UKC and AKC hunting events. We have many Dual Champions as well as field winners over the years, and strive to better ourselves with each litter. We offer a 10 year health guarantee. Will ship anywhere. Paypal accepted.
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Dunbar, Kentucky 42219
Breeding quality gun dogs for over 24 years.
270-925-1429Send Email to Redwing KennelClick here to visit Redwing Kennel website
Testimonials (4)
Submitted by: Pat and Bruce Frankinburger on Jan 15, 2014We brought Dexter home to Indiana with us in May of 2013. He was 8 weeks old. We bonded on the 6+ hour drive home, and he's become a vibrant member of the family ever since. Friendly, energetic, and biddable, Dex is the best!
Submitted by: Ginger Shaw on Jul 25, 2013I have had setters for more than 45 years, and purchased my last "baby" from Redwings in 2005. He was soooooo easy to train. We NEVER had an accident in the house, even when he was only 6 weeks old. I brought home this little 6# ball of fur that is healthier, smarter, and loves to "work" as my medical companion. I have a neuro disorder, diagnosed in 2010. Brogan senses the changes, and will either not leave my side, or goes to get someone to help if he thinks I need it. He is a bit mommy obsessed, but who says that HAS to be bad? :) He has put himself between me and danger more than once without questioning it.
Jean's care of her pups cannot be doubted. The heated barn they were in was clean, plenty of food and fresh water, and she has people who help socialize the pups until they go to their new homes. I can't wait to add another one of her pups to my family once we move :)
Submitted by: Larry on Mar 24, 2013I have purchased 4 Setters from Jean over a 20 year span. I have loved all my Redwings they are wonderful hunters, companions and family members. I raised my children with them and now my children own Redwing setters. They live long and healthy. They are easy to train. I have owned 2 reds and 2 red & whites. One daughter has Redwing reds and the other daughter has Redwing Red & Whites.
My grandson is starting to hunt with me maybe a Gordon this time as a present for him.
Thank you
Submitted by: John Koshoffer on Sep 20, 2011My boy, Squire, was fromJean's Redwing kennel and is both my hunting partner and a champion show dog in both UKC and AKC.
What more do you want??


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Forum > Raves and Rants >
does anyone know how to fix this? I loaded the chictopia widget onto my wordpress blog (amyswalkinwardrobe.wordpress.com) and it looks great, but still says i have '0' fans when i actually have 5. And obviously 5 sounds way better than 0 hahaha
would love any advice on this, really bothers me
It takes a while for it to update. My widget is usually behind by a couple days.posted about 4 years ago
thanks for letting me know. Mine always says 0 :( Strange thing is, it says the correct number when i view from my Ipad, but 0 from a PC. craziness.posted about 4 years ago
How strange! Have you tried deleting the widget code and typing it back in?posted about 4 years ago
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Samantha Peter and Robbie Pannet were married during a ceremony Tuesday, June 5, 2012, at the city park gazebo in Cannelton. Chris and Karen Zuckschwerdt officiated the ceremony.
The bride is the daughter of Cheryl and Bill Peter of Leopold and the late Larry A. Peter, and the granddaughter of Eva Knable. She is a full-time student at Sullivan College in Louisville, Ky.
Mr. Pannet is the son of Dave F. and Patty Pannet of Tell City and the grandson of Jeanette Pannet and the late Dave A. Pannet. He is enlisted in the U.S. Army and stationed at Fort Benning, Ga.
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Hundreds of New Zealand cancer patients would not have died from the illness if they were faced with the same battle in Australia, according to a leading researcher.
Studies published in the New Zealand Medical Journal today reveal death rates from cancer in New Zealand patients were 10 per cent higher than our Australian counterparts.
Researchers studied and compared the cancer incidence and death rates in both countries for the years 1996-97 and 2006-07.
The report found for 11-year timeframe, cancer-specific death rates decreased in both countries and cancer mortality fell in both countries. But there was no change in the difference between the death rates New Zealand and Australia, which remained remained 10 per cent higher in New Zealand.
In the publication's editorial, University of Otago professor David Skegg said New Zealand experienced more deaths from cancer that would be expected, and pointed to worrying figures.
"If the Australian rates had applied here, there would have been 616 fewer cancer deaths each year among New Zealand women, and 215 fewer in men. The majority of excess deaths were in people under 70 years of age," he said.
Almost 8000 New Zealanders die of cancer each year on average, figures from a 2000 to 2007 study show.
The main cancer sites responsible for the overall excess mortality were colorectual cancer in both men and women, and lung and breast in females. Every year, about 350 Kiwis die from bowel cancer who would not if the Australian rates were applied here.
"The higher mortality from all cancers combined cannot be attributed to higher incidence rates, and this suggests that overall patient survival is lower in New Zealand," Skegg said.
He said it was encouraging to see between 1990 and 2007, the overall cancer death rate declined by about 24 per cent in men and 20 per cent in women.
The study, A comparison of cancer statistics in New Zealand and Australia: 1996-2007, concluded differing cancer death rates was partly due to lifestyle and different ethnic populations, as well as New Zealanders presenting with more advanced cancers and having harder access to some treatments.
"Until we know the relative contributions of these factors, it will be difficult for New Zealand to plan interventions in the future which have a good chance of lifting our cancer survival rates to those of our closest neighbour. The collection of clinical stage on all new cancer registrations would provide the base information required," the study said.
Between 2000 and 2007, the study showed colorectum and anus cancer rates were 35 per cent higher in New Zealand women compared with Australia, and 24 per cent high in Kiwi men.
Meanwhile, lung cancer deaths were 20 per cent higher in New Zealand women compared with their Australian counterparts, but between men the rate was similar.
New Zealand's breast cancer mortality rate was almost 19 per cent higher during that period.
Both countries have population-based screening programs, starting in Australia in 1991 and in New Zealand in 1998, offering twice-yearly mammographic screening.
It has been estimated in the New Zealand programme, deaths were reduced by as much as 40 per cent in women participating in the screening.
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Sunday, 26 June 2016
Last updated 1 day ago
May 20 2013 | 12:23pm ET
China's Fosun International hopes to raise US$1 billion for its second dollar-denominated private-equity fund.
The new China Momentum Fund will invest in foreign companies seeking to expand in China. There will be a particular focus on Europe, in an effort to profit from depressed prices due to the economic crisis.
China Momentum will hold its first close next month at about US$300 million. Among the investors is Prudential Financial, Fosun's partner on its last fund, which raised US$600 million. The new fund will have priority over Fosun's existing funds in selecting investments.
Fosun aims to have only about 10 investors in the new fund, primarily pension funds, high-net worth investors and institutional investors from outside China.
Jun 16 2016 | 4:19pm ET
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Jun 17 2016 | 1:05pm ET
Over the past year, a large portion of the financial services industry has awakened...


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Weather News
Dry season floods Muchea with sheepOlivia Garnett, Tuesday July 2, 2013 - 17:18 EST
Stock agents say the recent dry spell in the eastern wheatbelt has been the main driver for 21 730 sheep and lambs being yarded at today's sale at Muchea.
It's the most sheep sold at Muchea in one day since May 2011, where 23 948 went under the hammer.
The lack of feed and water in northern and eastern parts of the wheatbelt has forced many producers to sell their animals.
One farmer from Dalwallinu sent 1600 sheep from his single property to today's sale, a sobering sign that producers are making tough management decisions.
Despite the high numbers, prices held well and quality was relatively good.
"I've just done an average on what I've bought and I've found that prices are very similar to prices at Katanning last week," said Murray French from Westcoast Livestock in Albany.
The WA Minister for Agriculture and Food Ken Baston says he's determined to create a diversification of markets for the sheep industry and he's been talking about those opportunities with producers at today's sale.
For many in the sheep industry those new markets couldn't come soon enough.
Reduced opportunities in the live export market in recent years, subdued demand from the processors, poor prices and now dry seasonal conditions have farmers offloading a large number of livestock at today's Muchea sale.
"One of my big issues is marketing, if you create the price where the producer can make a profit then I believe that's the best way we can build up the sheep flock in Western Australia."
Mr Baston is focussing on South East Asia, China and other emerging markets like Vietnam.
WA's sheep flock has dropped from 25 million to 14.5 million in six years, a level not experienced since the 1950's.
Mr Baston says today's yarding at Muchea of over 20 000 sheep should be the norm.
"We need to get back to that and show profitability in sheep."
He's concerned WA could lose its grip on overseas markets if numbers don't improve.
"I intend to travel and help to establish those markets for Western Australia, we've got to remember that on the same time zone we have 60 per cent of the world's population sitting on our doorstep."
Ron Dewar from Gingin was at the Muchea sale to sell some of his sheep which fetched for $104.40 each, higher than he expected.
"We expected a drop of about $10 considering the number of lambs that were in," Mr Dewar said.
"There were a couple of keen buyers that made the difference... it's a game of chance this farming."
"We're crying out for rain now, further inland I don't think they've had a break to the season at all."
"We're not badly off compared to a lot of the other poor devils."
One of the state's big sheep farmers and traders Peter Boyle from York took home about 650 sheep, a modest day considering he usually only gets the chequebook out for upwards of 1000.
He says the seasonal conditions are the main reason why there were so many sheep at today's sale.
"The dry season is starting to really bite, people are having to get rid of sheep because pasture is disappearing."
Mr Boyle says it's hard for producers to move ewes on when most are lambing.
"Hopefully they can either find agistment or they'll just have to feed them and carry them through."
The truckload of sheep Mr Boyle bought today are destined for the live export trade.
"Livestock Shipping Services have two boats going and Emanuel Exports have one," he said.
More breaking news
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Wintry weather across the nation
Cold snap across the nation reminds Australians we do have a winter.
Lorinna residents still relying on barge after floods cut road access
Almost three weeks after the devastating floods in north-west Tasmania, there is still no road access for the small community of Lorinna.
Temperatures plummet in southern Queensland as Brisbane shivers through coldest morning of year
Brisbane has shivered through its coldest morning of the year with temperatures dropping to below zero in parts of southern Queensland.


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At right: The Flip and Kale's Treenut attraction, which recently celebrated its populated anniversary
Nobody knows what the target audience of Flip and Kale's Treenut is. It once had stuff to do, but some stuck-up paralegals had it taken away to make it harder for kids to get exercise.
Run from Princess Mode
© copyright 2011–2016
A Pants Aflame production


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Utilities stocks (XLU -1.2%) are under heavy selling pressure as interest rates soar: EXC -1.7%, EIX -2%, FE -1.9%, PCG -1.5%, ED -1.4%, D -1.2%, PEG -1.3%, DUK -1.2%, NU -1.7%, SO -1%, NEE -1%, NRG -1.3%, SRE -1.6%, ETR -2.3%, XEL -1.7%, AEP -1.3%.
Utilities stocks (XLU -1.2%) are under heavy selling pressure as interest rates soar: EXC -1.7%,...
Jul 5 2013, 10:58 ET | About: Utilities Select Sector SPD... (XLU) | By: Carl Surran, SA News Editor
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Comments (7)
The Long Tail of Finance, contributorComments (1651) | + Follow Following - Unfollow
Nothing said about number of shares available. Just the number of owners of shares wanting to sell is greater than number of buyers at a given price point, forces prices down until buyers say OK, we'll buy shares now at that price. Simple economics - supply (# selling) and demand (#buying) determines price at which shares change hands. RSR
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Buffalo Praised For NHL Draft Yahoo Praised For Draft Streaming Show Debut Vegas NHL Team Sets Sights On Practice Facility Warriors Not In Need Of Drastic Changes Excel Sports Reps Six First-Round Picks Predators' Freeman Sues Team, Fellow Owner NHL Prospects Coming From Warm-Weather Cities NBA Draft Overnight Lowest Since '12 Foley Shifts Focus To Hockey Operations 76ers Excited For Ben Simmons
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The payrolls of each NBA team were listed in the Newark STAR-LEDGER by Dave D'Allesandro. The top three, in millions: Bulls, $61.87M; Knicks $53.70M; Magic, $42.48M (Newark STAR-LEDGER, 11/6)....The impact of the NHL's six "prominent" unsigned Group II free agents on the early part of the league's season is examined by SI's Michael Farber. Fox/MSG hockey analyst John Davidson: "I understand business, but the [1994-95] lockout beat us up, and I'd put these players' being out only a little behind that" (SPORTS ILLUSTRATED, 11/10 issue)....The Corel WTA Tour's Kremlin Cup in Moscow will increase its '98 prize money to over US$1M, making it the eight WTA Tour event to top the million dollar mark in prize money. This year's Cup set an attendance record with 59,397 for the week, more than triple its previous best of 17,000 (Corel WTA Tour).


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London Chocolate Spa day
A decadent combination of the popular chocolate rub and wrap for the body combined with a luxury chocolate pedicure.
This will all be served with a delicious cup of hot chocolate, and will last for two and a half hours.
If you love chocolate, you will adore the Savana Chocolate Indulgence Experience
Your amazing day will include:
A Chocoholics delight...
-You will start with the luxurious skin conditioning body wrap featuring warm oils. fine brown sugar and pure cocoa
-You will then receive a relaxing full body massage with warmed cocoa butter to sooth the spirit and leave the skin feeling smooth and smelling yummy.
-Next, to pamper those little toes you will receive a warm milk soak and hot vanilla and brown sugar scrub
-Finally you will experience a cocoa butter foot masque for super hydration
-This will all be served with a delicious cup of hot chocolate.
This delicious experience will last for two and a half hours.
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This gift is valid for 8 months from date of purchase. This certificate is non-transferable Please book in advance and arrive 15 minutes early for your appointment. Not to be used in conjunction with any other offers. This experience is for one person only. If you choose to have your pedicure with toe polish, please ensure you bring open-toed sandals or flip flops to wear after your treatment. This experience will last for two and a half hours.
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Octavia Nasr is CNN's Senior Editor of Arab Affairs and appears often on that network as an expert and commentator.
Here is what she tweeted upon hearing of the death of Hezbollah spiritual leader Muhammad Hussein Fadl-Allāh:
"beyond imagination."
(h/t DeJerusalem who provided the screenshot; Nasr's tweets are not public)
Lee Diamond – Sarona and Orlando Revisited
3 hours ago


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Church of Good Shepherd flock upset
- Details
The Parochial Church Council (PCC) of Bishopston and St Andrews have proposed the closure of the Church of the Good Shepherd Site
At the Sunday service on the 17th April, it was formally announced that the whole of the Church of the Good Shepherd (COGS) site, on Bishop Road, is to be sold off.
Students Moving Out
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STREET SCENE GROUP - NEWSLETTER – May 2016
Cricket Club - Frequently Asked Questions
- Details
Following the Residents' Meeting at the Cricket Club in April, a list of Frequently Asked Questions about the Cricket Club, cricket matches and the floodlights has been compiled by Matt Jones, Bristol City Council Neighbourhood Officer.
The responses have been provided by various of the stakeholders involved.
Bishopston Open Gardens Event: June 19th 2016
- Details
Come and see some beautiful local gardens on the afternoon of Sunday, June 19th. Full details of venues:
Friends of the Memorial Ground: Update, May 24th '16
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The Friends of The Memorial Ground group is being set up to ensure a fitting future for the Memorial Ground. The Friends will promote a positive, community use for the site, in keeping with its history and what it is for.
Memorial Ground Survey launched - please give your views!
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You are invited to contribute your views on the future of the Bristol Rugby Memorial Ground.
Bishopston,Cotham & Redland Neighbourhood Partnership News, May '16
- Details
Details of the Combined Forum where you can meet your new councillors and help plan and prioritise new projects, a listing of local events like The Free Harmony Picnic in St Andrew's Park, Sun 22nd May 11.30am onwards, Meet the Bees on Gloucester Road Sat June 11th etc can all be found in the latest BCR NP newsletter here
A New Library in Bishopston
- Details
This information comes from the BCC Libraries Team. Later this year, Cheltenham Road Library will be closing its doors and moving up the Gloucester Rd to a new venue in the former swimming baths site in Bishopston.
We'd love to know what you would like to see happen there.
The Bishopston Society's response to planning application for 2, Ashley Down Road
- Details
The Bishopston Society is concerned about the height and bulk of this proposal at second floor level in close proximity to the rear of 389 and 391 Gloucester Road. We consider that the building as it currently stands at 2 storeys high is the correct height.
We recommend refusal.
Friends of Horfield Common news May 2nd '16
- Details
You can see the programme of events happening through the summer - including new music evenings at the Ardagh, wildlife events, activities for young people during half term, and information about the cafe, ongoing weekly groups and new projects here


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Dell World: Dell warns of cyber sleeper cells targeting critical infrastructure
SecureWorks CTO warns of new strains of malware targeting governments and energy firms
AUSTIN: Dell SecureWorks chief technology officer, Jon Ramsey has warned cyber espionage tools like Flame are likely precursors to more serious Stuxnet level cyber attacks. Ramsey told V3 that Dell...
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We take our usual look back over what Microsoft has been up to this week
Brexit spells tough times ahead for the UK's tech industry
Good luck to all involved. You will need it
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