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A fluorescent labelling composition comprises a linear polysaccharide backbone molecule having a plurality of target-binding molecules, such as antibodies or nucleic acids, attached at spaced-apart intervals thereon. Each of the target-binding molecules, in turn, includes a multiplicity of fluorescent dye molecules bound thereto. In this way, fluorescent signal introduced to a single target-site on a solid phase surface may be increased without loss of binding activity.
1. A method for labelling a target substance in or on a solid phase, said method comprising:
providing a labelling composition including:
(a) a linear polysaccharide having a molecular weight of at least 500 kD;
(b) a plurality of molecules which specifically bind to the target substance attached to spaced-apart locations along the linear polysaccharide, and
(c) a multiplicity of fluorescent dye molecules bound to at least some of the target-binding molecules; and
exposing the labelling composition to the solid phase.
2. A method as in claim 1, wherein the solid phase is selected from the group consisting of a polymeric surface, an inorganic surface, and a tissue section.
3. A method as in claim 2, wherein the labelling composition is exposed to a solid phase surface which has previously been prepared by:
providing a solid phase surface having a specific binding substance specific for the target substance immobilized thereon; and
exposing the solid phase to a biological sample suspected of containing the target substance, whereby target substance is bound to the solid phase; and
wherein the labelling composition exposing step comprises exposing the labelling composition to the solid phase.
4. A method as in claim 1, wherein the linear polysaccharide is a dextram.
5. A method as in claim 4, wherein the linear polysaccharide is dextrans having a molecular weight of at least 1000 kD.
6. A method as in claim 1, wherein the target binding molecules are selected from the group consisting of antibodies, antibody fragments, antibody analogs, antigens, nucleic acids, hormones, hormone receptors, enzymes, enzyme substrates, avidin, and streptavidin.
7. A method as in claim 1, wherein the linear polysaccharide is dextrans having a molecular weight of at least 1000 kD and the target binding molecule is an antibody.
8. A method as in claim 1, wherein the fluorescent dye molecules are cyanine dyes.
9. A method as in claim 8, wherein the cyanine dyes are arylsulfonate cyanines.
10. A method as in claim 9, wherein the arylsulfonate cyanine is Cy5.
11. A method as in claim 1, wherein each target-binding molecule has from 1 to 10 fluorescent dye molecules covalently bound thereto.
12. A method as in claim 1, wherein the linear polysaccharide is dextrans having a molecular weight of at least 1000 kD, the target binding molecule is an antibody, and the fluorescent dye is an arylsulfonate cyanine.
The subject matter of the present application is related to that disclosed in each of the following U.S. patent applications which are being filed on the same day: U.S. Ser. Nos. 08/522,048; 08/522,434; 08/521,615; 08/522,435, the full disclosures of which are incorporated herein by reference.
1. Field of the Invention
The present invention relates generally to compositions and methods for detecting target substances in biological samples. More particularly, the present invention relates to labelling compositions comprising a multiplicity of fluorescent dye molecules bound to a plurality of binding molecules which in turn are bound to a linear polysaccharide backbone.
Many methods are known for detecting specific biological substances in solution and on solid phase supports. Of particular interest to the present invention are immunochemical methods where binding of a detectable label to a target substance in a biological sample is mediated by a specific binding substance. For example, a labelled specific binding substance, such as a fluorescently-labelled antibody, which recognizes the target substance (e.g., an antigen or hapten) is exposed to a biological specimen suspected of containing the target substance, and presence of the target substance is confirmed by binding of the label. Similar techniques may be performed using nucleic acid hybridization methods, where nucleic acid sequences of interest may be detected with a labelled polynucleotide which recognizes its complementary sequence in the nucleic acids of the biological specimen. In both cases, if present, the target substance will thus bind to the fluorescent label, allowing detection based on fluorescent emissions of the label.
Fluorescent labels offer a number of advantages compared with other common assay labels, such as enzyme labels. The detection of fluorescent labels is simple and does not require the addition of a substrate as is common with many enzyme labels. The signal generated by fluorescent label is localized, permitting the detection of multiple analytes in different reaction zones within a single reaction region or chamber. Thus, fluorescent labels allow simplified protocols for the detection of multiple analytes where only the final detection step must be performed separately for different analytes.
Of specific interest to the present invention, a class of cyanine fluorescent materials, referred to as "arylsulfonate cyanine fluorescent dyes," having particularly desirable properties has been developed. The arylsulfonate cyanine fluorescent dyes have high extinction coefficients (typically from 130,000 L/mole to 250,000 L/mole, good quantum yields, fluorescent emission spectra in a range (500 nm to 750 nm) which is outside of the autofluorescence wavelengths of most biological materials and plastics, good solubilities, and low non-specific binding characteristics. A particular arysulfonate cyanine fluorescent dye, designated Cy5, has the following structure: ##STR1##
Despite these excellent properties, arylsulfonate cyanine fluorescent dyes suffer from certain limitations. In particular, these dyes have a relatively narrow Stokes shift which results in significant overlap between the excitation and emission spectra of the dye. The overlap of excitation and emission spectra, in turn, can cause self-quenching of the fluorescence when the dye molecules are located close to each other when excited. Such self-quenching limits the number of arylsulfonate dye molecules which can be conjugated to a single antibody molecule for use in immunoassays. In the case of Cy5, an exemplary arylsulfonate cyanine fluorescent dye, the Stokes shift is 17 nm (which is the difference between an excitation wavelength of 650 nm and an emission wavelength of 667 nm). Optimal fluorescent yield is obtained when from two to four Cy5 molecules are conjugated to a single antibody molecule. The fluorescent signal output drops rapidly when more than four dye molecules are conjugated to a single antibody molecule. The inability to conjugate more than four dye molecules to individual antibody molecules significantly limits the sensitivity of immunoassays using Cy5-labelled antibodies and other binding substances.
The direct attachment of fluorescent dye molecules to carrier molecules, such as dextran, has been proposed as the technique for enhancing fluorescent output. Such carrier molecules would typically contain one or a few antibody molecules, where the binding of each antibody to a target substance will carry a number of fluorescent dye molecules. Attachment of arylsulfonate cyanine dyes to such carrier molecules, however, would not be expected to significantly increase the level of fluorescent emission because of the self-quenching density of the individual dye molecules, as described above. Moreover, the binding capacity of many prior fluorescent dyes/carrier molecules complexes has been limited, due both to a limited number of antibody molecules present in the complex and to steric hinderance of the antibodies from the carrier molecule.
For these reasons, it would be desirable to provide improved fluoresceht labelling compositions for use in the detection of target substances in biological samples. Such compositions should be useful in all circumstances where fluorescent labelling complexes generally find use, including solid phase immunoassays, immunohistochemical staining, flow cytometry, and the like. It would be particularly desirable to provide improved arylsulfonate cyanine dye compositions comprising conjugates having multiple dye molecules with reduced self-quenching and relatively high target-binding capacities. Such compositions should continue to display the low non-specific binding, high solubility, and other desirable characteristics associated with previous arylsulfonate cyanine dye formulations.
2. Description of the Background Art
Arylsulfonate cyanine fluorescent dyes are described in Mujumdar et al. (1993) BIOCONJUGATE CHEMISTRY 4:105-111; Southwick et al. (1990) CYTOMETRY 11:418-430; and U.S. Pat. No. 5,268,486. Cy5 is described in each of the references and is commercially available from Biological Detection Systems, Inc., Pittsburgh, Pa., under the tradename Fluorolink™ Cy5™. Fluorescently labelled polysaccharides, such as dextrans, are described in Globe et al. (1983) ANAL. BIOCHEM. 130:287 and deBelder et al. (1973) CARBOHYDRATE RES. 30:376.
The present invention provides compositions and methods for highly sensitive fluorescent detection of target substances and biological samples. The compositions are fluorescent labelling compositions comprising a carrier or backbone molecule having a plurality of target-binding molecules bound thereto. Each target-binding molecule, in turn, has a multiplicity of fluorescent dye molecules bound thereto. The backbone molecule is a linear polysaccharide, such as a high molecular weight dextran, and the target-binding molecules are antibodies, nucleic acids, or the like, bound at spaced-apart locations along the linear polysaccharide. Such compositions are particularly suitable for incorporating arylsulfonate cyanine fluorescent dyes, such as Cy5, where they provide substantially enhanced fluorescent emission levels (signal amplification) with reduced self-quenching.
The compositions of the present invention may be advantageously employed in virtually any assay or labelling procedure where fluorescent label is to be introduced to a target substance via specific binding, including solid phase and other heterogenous immunoassays, immunohistochemical staining of biological specimens, flow cytometry, and the like. The compositions will be particularly useful in performance of solid phase assays where the labelling composition is exposed to a solid phase surface having the target substance bound directly or indirectly thereto. An especially preferred use is in two-site or "sandwich" solid phase assays where the solid phase surface is derivatized with a binding substance specific for a target substance suspected of being present in a liquid sample. After exposure to the liquid sample, the solid phase is washed and exposed to the labelling composition of the present invention which binds directly or indirectly (e.g., through the intermediate binding of biotin and avidin/streptavidin) to the immobilized target substance. The fluorescent label is then detected on the solid phase surface by exposure to excitation energy within a localized reaction zone on the surface.
FIG. 1 is a schematic illustration showing the structural relationship of the polysaccharide backbone, the target-binding substance, and the fluorescent label in the fluorescent labelling compositions of the present invention.
FIGS. 2A and 2B illustrate a method according to the present invention using the fluorescent labelling composition of FIG. 1 for detecting target substances in two discrete reaction zones on a solid phase surface.
FIG. 3 illustrates the use of specific binding substances for capturing a target substance onto a solid phase according to the method of the present invention.
Fluorescent labelling compositions according to the present invention comprise conjugates 10 (FIG. 1) including a backbone or carrier molecule 12, a plurality of target-binding molecules 14, and a multiplicity of fluorescent dye molecules 16 attached to each of the target-binding molecules. The backbone or carrier molecule 12 will comprise a linear polysaccharide, usually consisting of a single linear polysaccharide molecule. Conjugate 10 will include at least two target-binding substances 14, usually including at least four target-binding substances, and preferably including from five to 20 target-binding substances, more preferably from six to 15 target-binding substances. Each of the target-binding substances 14 will preferably include at least one fluorescent dye molecule, more preferably including at least two fluorescent dye molecules, and often including three or more fluorescent dye molecules. For many fluorescent dyes, the upper limit on the number of dye molecules will be 5, 10, or even higher. In the exemplary case of arylsulfonate cyanine fluorescent dyes, however, the optimum number of fluorescent dye molecules will be from 2 to 4 dye molecules per antibody binding substance 14.
The compositions and methods of the present invention may be used for labelling and detecting a wide variety of target substances in various biological samples. The term "target substance" is used herein as defined as a molecule, compound, or other composition which is suspected of being present in a biological sample or on a solid phase surface, as described hereinafter. The target substance will be detectable through binding through a target-binding substance as described below, and will usually be a biological molecule, such as a polypeptide, protein, carbohydrate, or nucleic acid, and may thus be associated with a particular biological, pharmacological, genetic, or biochemical property or set of properties. A non-exhaustive list of exemplary analytes is set forth in U.S. Pat. No. 4,366,241, at col. 19, line 7 through col. 26, line 42, the disclosure of which is incorporated herein by reference.
Target substance may be present in or separated from a variety of biological specimens, such as blood, serum, plasma, urine, cerebral fluid, spinal fluid, ocular lens liquid (tears), saliva, sputum, semen, cervical mucus, scrapings, swab samples, and the like, of the type frequently employed in the diagnosis in monitoring of diseases and therapeutic treatments. In addition to such medical samples, the compositions and methods of the present invention may be used to detect target substances in industrial, environmental, and food samples, such as water, process streams, milk, meat, poultry, fish, condition media, and the like. In certain circumstances, it may be desirable to pretreat the sample, such as by liquification, separation, solubilization, concentration, filtration, chemical treatment, or a combination of such treatment steps, in order to improve the compatibility of the sample with the remaining steps of the assay, as described hereinafter. In particular, it will frequently be desirable to pretreat a sample in order to permit reaction with a solid phase surface in order to capture a target substance on the surface prior to detection with the fluorescent compositions and methods of the present invention. The selection and pretreatment of biological, industrial, and environmental samples prior to immunological testing is well know in the art and need not be described further.
The term "target-binding substance" as used herein will refer to any macromolecular compound having spatial and polar features which cause it to bind specifically to another compound or feature in a complex biological material. Such target-binding substances useful in the present invention will be selected or prepared to specifically bind to particular target compositions, such as the target substances defined above. Natural specific binding pairs which may provide the target-binding substances include antibodies and antigens, lectins and carbohydrates, hormones and hormone receptors, enzymes and enzyme substrates, biotin and avidin/strep avidin, vitamins and vitamin-binding proteins, complementary polynucleotide sequences, drugs and receptors, enzymes and reaction products, enzymes and enzyme inhibitors, apoproteins and cofactors, immunoglobulins and receptors, organisms and receptors, growth factors and receptors, chelating agents and metals, and the like. Biotin and avidin, including biotin derivatives and avidin derivatives such as streptavidin, may also be used as intermediate binding substances in assay protocols employing complex binding sequences. For example, antibodies may be labelled with biotin ("biotinylated") and used to bind to a target substance previously immobilized on a solid phase surface. Fluorescent compositions according to the present invention employing an avidin target-binding substance may then be used to introduce the fluorescent label. When no natural specific binding substances exist (as will usually be the case), a specific target-binding substance may be prepared. For antigenic and haptenic target substances, antibodies may be prepared by well-known techniques. For polynucleotides, complementary polynucleotides, including both DNA and RNA molecules, may be prepared by well known synthesis techniques.
Suitable backbone or carrier molecules for incorporation into the labelling conjugates of the present invention will be linear polysaccharides having molecular weights above 500 kD, preferably above 750 kD, and more preferably above 1000 kD. Exemplary linear polysaccharides include dextran, FICOLL�, and the like. Particularly preferred is the use of dextran molecules having molecular weights above 100 kD, preferably above 1500 kD, and frequently 2000 kD or higher. A preferred linear polymer is dextran having a molecular weight of 2000 kD.
The compositions of the present invention may incorporate virtually any fluorescent label which may be covalently or non-covalently (preferably covalently) attached to the binding substance. In the case of both protein and nucleic acid target-binding substances, suitable fluorescent labels include fluorescent dyes, fluorescein, Texas Red, and the like. In both cases, the preferred fluorescent label will be an arylsulfonate cyanine fluorescent dye, such as those described in U.S. Pat. No. 5,268,486, the full disclosure of which is incorporated by reference. Particularly preferred is the arylsulfonate cyanine fluorescent dye designated Cy5, in that patent.
Attachment of the fluorescent label to the target-binding substance may be covalent or non-covalent, preferably being covalent. The manner of covalent attachment will depend on the precise nature of both the label and the target-binding substance. When the target-binding substance is a polypeptide or protein, such as an antibody, the fluorescent label may be covalently bound to it through a variety of moieties, including disulfide, hydroxyphenyl, amino, carboxyl, indole, or other functional groups, using conventional conjugation chemistry as described in the scientific and patent literature. Alternatively, antibodies may be biotinylated by known techniques (see Wilchek and Bayer, (1988) ANAL. BIOCHEM. 171:1-32) and linked to the fluorescent label via avidin molecules (where the fluorescent label-avidin conjugate may comprise the fluorescent labelling composition of the present invention).
Covalent binding of the fluorescent label to a polynucleotide target-binding substance may be effected through a variety of moieties, including aldehyde, ketone, isothiocyanate, imidate, inosine, acyl, and alkyl, using conventional conjugation chemistry, while derivatization with biotin is taught in numerous references. (See Leary et al. (1983) PROC. NATL. ACAD. SCI. USA 80:4045-4049; WO86/02929; EP063 879; Langer et al. (1981) PROC. NATL. ACAD. SCI. USA 78:6633-6637; and EP2009 996). Numerous other references are available which teach methods for binding fluorescent labels to nucleic acid molecules.
Exemplary techniques for binding arylsulfonate cyanine fluorescent dye labels to antibodies and other proteins are described in U.S. Pat. No. 5,268,486, the full disclosure of which has previously been incorporated by reference. Techniques for linking the preferred Cy5 fluorescent label to both antibodies and nucleic acids are described in a technical bulletin identified as Cat. No. A25000, published by Biological Detection Systems, Inc., Pittsburgh, Pa.
The target-binding substances will be bound to the linear polysaccharides using known procedures for linking proteins and/nucleic acids to carbohydrates. For example, Manube et al. (1983) BIOCHEM. BIOPHYS. RES. COMM. 115:1009, describes conjugation of the anti-cancer agent, mitomycin, to a monoclonal antibody conjugated to dextran. The dextran enhanced the cytotoxic effect of the drug by crosslinking the antibody. Scheffler et al. (1972) J. BIOL. CHEM. 247:18, developed a method to link polynucleic acids to FICOLLi� and dextran to study the activity of DNA polymerases. McMaster et al. (1975) CELL. IMMUNOLOGY 20:42-53, prepared dinitrophenolficoll to monitor hapten specific B cell immune response. There are a variety of chemical approaches to effect linkage of the target binding substance to the linear polysaccharides. Crosslinking reagents directed towards carboxyl, sulfhydryl, and amino groups can be employed. Wong gives a comprehensive description in "Chemistry of Protein Conjugation and Crosslinking," CRC Press. The number of antibodies bound per polysaccharide molecule will depend on the molar ratio of antibody to polysaccharide (dextran) during the conjugation reaction.
The labelling compositions of the present invention will be prepared from the molecular components described above using the conjugation chemistries also described above. After preparation, the compositions will be separated and purified using conventional separation techniques, such as gel filtration, HPLC, and the like. Typically, the compositions of the present invention will be at least 90% pure by weight, preferably being at least 95% pure by weight, and usually being 99% pure by weight, or higher.
The fluorescent labelling compositions of the present invention will frequently be employed to detect target substances present on solid phase surfaces. As used herein, the term "solid phase surface" will include any biological or non-biological surface capable of supporting or capturing the target substance. When used in heterogenous immunoassays, the solid phase surface will typically be a plastic (e.g., acrylic), glass, membrane, or other non-biological surface having a target-binding substance previously immobilized thereon. Specific methods for immobilizing capture antibody of other binding substances are described in copending application Ser. Nos. 08/374,265 and 08/522,435 the full disclosures of which are incorporated herein by reference.
The solid phase may then be used to react with a biological sample to capture target substance which may be present therein. Such solid phase surfaces are frequently used in two-site immunometric assays which utilize two antibodies, each of which is present in excess over the amount of target substance in the sample. The relatively high concentrations of target-binding substance and of labeled target-binding substance assures that all target substance is captured and detected. A particular protocol for performing such two-site immunometric assays will be described below in connection with FIGS. 2A and 2B.
The fluorescent labelling compositions of the present invention will also be useful with biological solid phase surfaces, including tissue sections, intact cells, biological membranes, organslies, chromosomes, and the like. The compositions will also be useful with non-biological membranes which are prepared by the transfer of a desired target substance from a primary biological material, e.g., by blotting.
Referring now to FIGS. 2A and 2B, an exemplary two-site immunometric assay performed using the fluorescent labelling compositions of the present invention will be described. A solid phase surface 30 includes a first target substance 32 and an immunologically distinct second target substance 34. The target substances 32 and 34 may be any of the types of substances described above, including proteins, carbohydrates, nucleic acids, and the like. At least two types of fluorescent labelling conjugates 10' and 10" are exposed to the solid phase surface 30, typically in an aqueous solution which permits which permits conjugates to migrate until they recognize their specific target substance. Conjugate 10' includes target-binding substances 14 which are specific for the target substance 32, while conjugate 10" includes target-binding substances 14" which are specific for the target substance 34. After sufficient incubation time, typically from 60 seconds to 600 seconds, the conjugates 10' and 10" will bind to their respective target substances 32 and 34, respectively, as illustrated in FIG. 2B. After washing to remove non-bound labelling conjugate, the discrete reaction zones on the solid phase surface 30 may be separately interrogated with excitation energy having a wavelength selected to excite the particular fluorescent label 16 which is employed. While the same fluorescent label 16 will usually be employed in both conjugates 10' and 10", the physical isolation of the reaction zones in which the conjugates are bound permits separate detection without interference. Thus, the amounts of both target substance 32 and 34 may be independently measured on a single solid phase surface.
It will be appreciated that target substances 32 and 34 will typically be introduced to the solid phase surface 30 using specific binding substances, such as capture antibodies 36 and 38, respectively (FIG. 3), which have previously been immobilized on the surface. Such aspects of two-site immunometric assays are well known in the art
The following examples are offered by way of illustration not by way of limitation.
Materials and Methods
1. Preparation of Cy5-Antibody-Dextran Conjugates
To thiolate dextran, 150 μl of a 34 mg/ml solution of succinimydyl 6-[3-[2-pyridyldithio]-proprionamido]hexanoate (LC-SPDP) (Pierce #21651) in dimethylformamide (DMF) was added to a 5 ml solution containing 20 mg/ml aminodextran (Molecular Probes, #D-7145, 130 amines/dextran, 2000 kD M.W.) in phosphate buffered saline (PBS) pH 7.4. The reaction proceeded for 30 minutes at room temperature (RT), then the mixture was dialyzed overnight at RT against PBS. Greater than 95% of the amines in aminodextran were labeled with LC-SPDP during the reaction. The LC-SPDP-dextran was then purified on a Bio Gel� A5m column (1�100 cm). To remove the low molecular weight dextran fragments, only the fractions of the first peak were collected.
A 6.15 ml solution of anti-CKMM antibody (BiosPacific G31520) at 2.6 mg/ml in PBS pH 7.4 was mixed with 156 μl of 2.7 mg/ml succinimidyl 4-[N-malemidomethyl]cyclohexan-1-carboxylate (SMCC) (Pierce #22320) in DMF and was allowed to react for 30 minutes at RT. Unreacted SMCC was removed by purification on a Sephadexe G25 column.
The LC-SPDP-dextran was reduced by adding 156 μl of 0.5M dithiothreitol to 5.2 ml of 3.1 mg/ml LC-SPDP-dextran and incubating for 15 minutes at 2 RT. The dextran was then purified on a PD-10 (Pharmacia #17-0851-01) column in PBS pH 7.4.
Conjugation was achieved by mixing the reduced LC-SPDP-dextran with the SMCC-anti-CKMM and incubating overnight at RT. n-ethyl-maleimide (NEM) (Sigma #12828-7) was added at a final concentration of 1.0 mM to stop the reaction. The conjugate was then concentrated down to 6 ml in a Centricon Concentrator (Amicon #4211), and the buffer was exchanged to 0.1M sodium carbonate pH 9.3.
The anti-CKMM/dextran was labeled with Cy5 at a molar coupling ratio (antibody:dye) of 1:4 by adding 0.1 mg of Cy5 (Biological Detection Systems 1464C-1) to 1.3 ml of the concentrated conjugate. After 30 minutes at RT, the conjugate was purified on a PD-10 column to remove free Cy5, and then purified on a Bio Gel� A5m column (1�100 cm) to remove unconjugated antibody. Spectral analysis of the conjugate indicated 2.9 Cy5 molecules per antibody. Approximately 10 antibodies were linked to dextran assuming an average molecular weight of 2000 kD.
2. CKMB Assay Materials
Reaction wells having benzophenone bovine serum albumin (BSA)-biotin immobilized on the bottom were prepared as follows. Each reaction well was assembled by attaching a teflon ring on the surface of an acrylic disk obtained from the Germanow-Simon company, using double stick tape 415 from the 3M company. The dimensions of teflon ring were 9 mm inner diameter and 1.5 mm height. The dimensions of acrylic disk were 30�30 mm sides and 0.6 mm thick, on which four teflon rings were attached. The teflon rings were etched with CO2 to improve bonding of the rings onto the disk. Benzophenone BSA-biotin was then immobilized at 40 μg/ml by UV-curing for 10 minutes.
Streptavidin-monoclonal anti-CKMB antibody conjugate (SA-anti-CKMB) was prepared using heterobifunctional linking reagents, S-acetylthioglycolic acid N-hydroxysuccinimide ester (SATA, Pierce #26102), and Succinimidyl 4-(N-maleimidomethyl) cyclohexane-1-carboxylate (SMCC, Pierce #22320). Fifteen (15) molar excess of SATA (dissolved at 5 mg/mL in dimethylformamide (DMF) from Pierce) was reacted with 1 mg of antiCKMB at 0.74 mg/mL in PBS at pH 7.4 for 3 hours at room temperature. Also, 15 molar excess of SMCC (dissolved at 5 mg/mL in DMF) was reacted with 1.1 mg of streptavidin at 1.1 mg/mL in PBS for 3 hours at room temperature. Unreacted linkers were removed from both antiCKMB/SATA and streptavidin/SMCC reaction mixtures using Sephadex� G25M column (PD10 column from Pharmacia). The purified antiCKMB-SATA and streptavidin-SMCC were mixed at the protein molar ratio of 1:3. The conjugation reaction was initiated by adding 1M hydroxylamine to a final concentration of 100 mM and incubated over 18 hours at 4� C. The reaction was stopped by adding 100 mMN-ethylmaleimide (NEM) from Aldrich Chemical at a final concentration of 1 mM in the reaction mixture and incubating for 15 minutes at room temperature. After the incubation with NEM, the conjugation reaction mixture was concentrated to a final volume less than 2 mL using a Centricon-100 concentrator from Amicon. Then the concentrated mixture was purified using a Bio-Gel� A-5m column (1�100 cm) from Bio-Rad to isolate streptavidin-antiCKMB conjugate.
Cy5/anti-CKMM-Dx conjugate was prepared as described above.
SA-anti-CKMB dilution buffer was 0.1% BSA and 0.05% Tween� 20 in PBS, pH 7.4. Assay Buffer was 4% HSA, 150 μg/ml mouse IgG, and 0.05% Tween� 20 in PBS, pH 7.4. Wash Buffer was 0.2% Tween� 20 in PBS, pH 7.4.
3. CKMB Assay Protocol
SA-anti-CKMB solution (7.5 μg/ml) was added to SA-anti-CKMB dilution buffer. 50 μL of this solution was added into reaction wells and incubated for 1 hour at 37� C. Wells were washed by immersing the entire disk into wash buffer. The wash step was repeated 5 times in five different containers containing wash buffer. Excess wash buffer was removed from the wells using a micropipet. Samples (50 μl) containing CKMB was added into the reaction wells and incubated for 10 minutes at room temperature. Wells were washed by immersing the entire disk into wash buffer. The wash step was repeated 5 times in five different containers containing wash buffer. Excess wash buffer was removed from the wells using a micropipet. Cy5/anti-CKMM-Dx conjugate (50 μl) was added at the proper loading and incubated for 5 minutes at room temperature. Wells were washed by immersing the entire disk into wash buffer. The wash step was repeated 5 times in five different containers containing wash buffer. Excess wash buffer was removed from the wells using a micropipet. Wash buffer (50 μl) was added into each well, and the Cy5 signal read using a reader.
The results are shown in Tables 1 and 2, below. The results of Table 1 are from a CKMB assay run with 0 (control) and 100 ng/ml samples and Cy5-labelled anti-CKMM. Assays employing Cy5-anti-CKMM-dextran can have up to a six-fold higher fluorescent signal with positive CKMB samples.
The fluorescent signal with negative samples (resulting from non-specific binding of the Cy5-antibody, was not increased when compared with the dextran conjugate and native antibody. Consequently, the signal-to-background ratio is improved with dextran conjugates.
Table 2 illustrates data from CKMB assays employing Cy5-anbtibody-dextran conjugates with dextrans of varying molecular weight. Dextran (200 kD) maximizes the available signal.
TABLE 1______________________________________Monomeric Anti-CKMM-Cy5 vs. PolymericCy5-Anti-CKMM/Dextran Conjugates Loading [CKMB Fluorescence (mV)Conjugates (μg/ml) (ng/ml) Mean SD______________________________________Anti-CKMM-Cy5 10 0 3.00 0.19 100 19.13 0.86 40 0 3.98 0.41 100 20.47 1.54 80 0 5.03 1.11 100 20.94 1.85Anti-CKMM- 10 0 2.96 0.50Cy5/Dextran (2000KD) 100 59.67 1.06 40 0 2.45 0.11 100 82.90 5.90 80 0 2.98 0.09 100 124.83 5.89______________________________________
TABLE 2______________________________________Comparison of Cy5-Anti-CKMM/Dextran Conjugates in Various SizesCy5-Anti-CKMM/Dextran Loading [CKMB] Fluorescence (mV)Dextran Molecular Wt. (μg/ml) (ng/ml) Mean SD______________________________________70 KD 40 0 1.72 0.23 100 28.85 4.77 80 0 2.95 0.64 100 35.25 1.76500 KD 40 0 1.76 0.28 100 65.12 4.82 80 0 2.58 0.88 100 51.82 4.942000 KD 40 0 2.24 0.18 100 105.67 9.22______________________________________
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Live Flesh
In art history textbooks, Schiele is often and carelessly labeled an Expressionist, a description intended to distinguish his eroticism from the decorative eroticism of his mentor, Gustav Klimt. Klimt, of course, depicted lovers clasped to each other in intense erotic embrace. There is nevertheless something operatic about Klimt's lovers, as if they were figures in a myth. Like Tristan and Isolde they are caught up in the sweep of passion as the music swells around them. Sex is somehow meant to be transfigurative, a way of transcending the sweaty realities of the flesh depicted. Schiele's figures, by contrast, are raw, hairy and bony, their young bodies marked by erotic zones like maps of where to touch each other. Sex is what they live for, the essence of their lives. It is an end in itself, not a means for transfiguration. They can't keep their hands to themselves when they are together, and they can't keep their hands off themselves when they are alone. Masturbation is their default state.
In their leanness, Schiele's figures might be said to resemble those of Picasso's Blue Period. But Picasso's figures are gaunt because they are poor and needy, whereas Schiele's have no thought for eating, as their only hunger is for sex. They are like illustrations of a thesis of Sigmund Freud, Schiele's fellow Viennese, that human reality is essentially sexual. What I mean to say is that there is no art-historical explanation of Schiele's vision. Expressionism was certainly in the air in Mitteleuropa in those years. But his drawings look like nothing one would see by artists who belong to movements like Die Brücke ("The Bridge") or Der Blaue Reiter ("The Blue Rider"). The German Expressionists used heavy black outlines and were inspired by a vision of primitivism. My own view, hardly inspired, is that Schiele expressed what Freud describes in his central thesis about human nature and conduct--that from infancy on, sex relentlessly holds us in its grip. In Schiele's work we see what we know is repressed in the men and women painted by Edvard Munch, the artist I think Schiele is closest to in terms of achievement.
The reference to Freud is not an appeal to a Viennese zeitgeist by which Schiele's work might be unpacked, although I think it says something about Vienna before World War I that eroticism was the main artistic achievement of the Austrian capital's most original artist at the time. Rather, I mean to suggest that Schiele is likely to have known about Freud's views, whose Three Contributions to the Theory of Sex was published in 1905. Consider Schiele's extraordinary Mother and Child, another drawing from 1910. The theme of mother and child has a considerable history in Western art, from Venus and Eros to the Madonna and the Christ Child, but there is nothing to compare with Schiele's study. The mother is shown from behind, looking over her shoulder, gazing back with a flirtatious glance. Her body is curved in a particularly sexy way, giving a thrust to her hips and a saucy swing to her generous buttocks. She is nude except for her black stockings, and we can see the tip of one breast from the side. The child is sitting next to her, on the arm of the chair the mother leans into. One of his hands is pressed against her waist, which he appears to be kissing with the intense fervor with which he would be sucking at the breast, if that were anatomically possible. The other hand conveys the child's total absorption in the mother's flesh. It is very much as if they are lovers--hardly a posture that would have occurred to anyone had the idea of infantile sexuality not been in the air. Schiele had drawn pregnant females at a clinic with a certain obstetrical precision. But Mother and Child has a moral daring, and it expresses a psychological truth. Interestingly, the pair is surrounded by a kind of white aura, scrubbed onto the yellow of the paper, and the flesh itself is given life by the way the paint is swirled on, as in finger painting. The space where the buttocks join the thighs is punctuated by a dark cross as black as the mother's eye or hair.
Kneeling Semi-Nude, done in 1917, shows how little Schiele's style and vision had evolved over seven years. The naked upper body of the kneeling woman emerges from a voluminous frilled undergarment. She is intensely involved in palpating her left breast, holding it in her right hand while she probes above the nipple with the other hand. The round red nipple is fully exposed, and the woman is peering at it with such intensity that the celebrated male gaze of contemporary feminist theory appears by contrast to be a passing glance. Like all of Schiele's women, she is slender and beautiful, and her face is framed marvelously by tangled black curls. Lips, cheek and nipple are the only touches of red in the otherwise neutrally painted body.

subset: commoncrawl


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Yenidze, Dresden
10 Reviews
10 Reviews
"Yenidze is a strange name" - I thought when I first heard it. It looked like Georgian word. Actually it is old cigarette factory, the name comes from former Ottoman Empire town. It was built in 1909, using architecture of mosques. The production of cigarettes continued till 1953, later it was Dresden tobacco association, nowadays here is restaurant, place for different events.
I have seen it from longer distance, but I believe it is reachable by foot from old town.
When we arrived in Dresden (from Berlin), the train passed at this totally out of Baroque style building which does not match anything in the city. First you might think of the Russian orthodox legacy, or a stone-turned content of a fairy-tale. The latter is closer to the look – as the dome is not orthodox onion-shaped but straighter mosque-like.
But it is all a little different, and the building has a very interesting history.
For a start, it is called Yenidze. DIE Yenidze. As you know, we have articles in German… ;-) To me Yenidze had sounded a bit more masculine, like DER Yenidze… ;-)
The name refers to the old name of the town of Giannitsa in Northern Greece – which was Yenidze and means "New Town". It has been known for tobacco plantations for more than a hundred years. And there we are: This building in the style of a mosque was a cigarette fabric for many years, built from 1909 to 1912, run by the cigarette producer Hugo Zietz. The locals called the building "Tabakmoschee" – tobacco mosque.
Once Dresden produced 60 % of Germany's cigarette production. The best-known brand was Salem – even I as a non-smoker remember it, as my grandfather used to smoke them.
After the war it did not take long to repair the damage, production started again in 1947 already, and from 1953 the Yenidze became the central tobacco office of GDR. After the reunification the building was thoroughly restored and redeveloped, and turned into an office complex. During the restoration the glass dome got back its original colours that had been replaced by uni-coloured glass after the War.
Under this dome there is a restaurant now, and in the basement you find the Märchen GmbH, a cultural place where they perform fairy-tales and more (Märchen = fairy-tale). On their website you can find their ever changing programme.
The building once caused heavy controversy, and the architect who dared to design it – a guy named Hermann Martin Hammitzsch - was excluded from Saxony's conservative architects' society!
The Yenidze was the world's first reinforced concrete skeleton construction. The cleverest thing was to disguise the chimney stalk as a minaret, as they would not have allowed such an industrial chimney in the royal residence of Dresden at the time.
It is said the architect had the splendid Mameluk grave of Khair Bak in Cairo on his mind when he designed the Yenidze in this style-mixture of art-deco and moorish. The building has 600 windows. The dome is 20 metres high, and has a diametre of 17 metres.
The Yenidze is located near Marienbrücke, north of Neustadt, next to the Elbe river. You can spot it quite well from the many viewing towers in the city.
The green building you see next to it, just around the corner in Ostra-Allee, is Sächsische Zeitung which is Saxony's leading newspaper. Remember the name – I write for them as a freelancer ;-)
Kuppelrestaurant in der Yenidze
Weißeritzstraße 3
01067 Dresden
Phone 0351-490 59 90
Fax 0351-490 59 92
Website: http://www.kuppelrestaurant.de/index1.html
Email: firstname.lastname@example.com
Open daily 11am - 12midnight
1001 Märchen GmbH
Weißeritzstraße 3
01067 Dresden
Phone 0351- 495 1001
Fax 0351 - 495 1004
Website: http://www.1001maerchen.de/
Email: email@example.com
The Yenidze is one of Dresden's more unusual buildings and certainly adds to the already impressive skyline. Built in 1908 in the form of a mosque it was originally a cigarette factory and was named after the company and derived from a city name in what is today Macedonia. Its architect Martin Hammitzsch came up with the tobacco mosque motif at the suggestion of the company's owner, who was of Turkish descent. It was restored to its former glory in 1996 after some years of disuse and now is a multiuse facility housing offices, a restaurant with a fantastic view and a unique beer garden. But perhaps its most interesting aspect is in the basement where fairy tales are reenacted in the evening.
Jenidze is my favourite building (and beergarden!) in Dresden! The mosque-style building used to be a tobacco and cigarette factory. It was built between 1907 and 1909. The colourful dome today hosts a small theatre where actors and writers read out different fairytales in the evenings. Unfortunately we didn't have time for such an event, it would have been my favourite thing to do. Sounds soooo romantic. Anyway, we had a drink on the terrace of the building which is located right underneath the big dome inbetween all the little towers and spires. You get a nice view of Dresden Downtown from here but the nicest view you get when looking up: The dome of Yenidze!
The building that looks like a mosque with its huge pointed dome and minaret, but it is no mosque and has never been. Jenidze was a factory that produced cigarettes. It was built in 1907-1912 for the "tobacco king" Hugo Zietz. Due to the Turkish-Arab origins of tobacco (or so they thought) the owners and architect chose the Middle Eastern style, covered the factory hall with a dome and turned the chimney into a minaret.
The factory has long been given up. The building is now mostly occupied by offices and gastronomy. A part of it, including the hall underneath the dome. is used for events, fairy-tale readings, exhibitions and concerts.
The oriental dome is a landmark in Dresden's skyline. The best view is from the train - right after crossing the Elbe bridge between Bahnhof Neustadt and Hauptbahnhof, from where you have hopefully enjoyed the panorama of the old town, look to the opposite site not to miss Yenidze, which is located next to the train tracks.
This office mall was built in 1907-09 and was intended to look like a mosque in order to refer to the tobacco theme by Hugo Zietz who imported the smokes and made the packets of tobacco here. The building has an onion dome and minarets and colored chimneys. The dome is 66 feet high and the building has 600 windows. The building has had financial problems for years and now only has a disco in the basement.
It stands out on the horizon over by the Neustadt rail station
Yenidze is a former factory for cigarettes and was built in 1909 by Martin Hammitzsch in oriental style. The name come from a town called "Genissea" in the former Ottoman Empire. Today Yenidze houses offices, a discotheque and a restaurant.
This building resembles a mosque with its dome-like cupola, though it has been a factory in former times. The architecture is interesting. It serves as a cultural centre - inside the cupola, cultural events, readings etc. take place.
The old tabacco contor is a bit off the beaten path. I really think it's the most fascinating building in Dresden as it's so unusual. You seem to be in the orient there...
Nowadays its mainly an office building. Under the green cuppola you will find a restaurant and a theater, showing fairy tales....
Yenidze was an old cigarette factory - the building is in the form of a mosque with a stained glass dome
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LAS VEGAS – Muhammad Ali had his Leisure World, too.
He was so uncontested in his heavyweight lair, that he turned much of his career into a world tour. Richard Dunn, Ruud Luebbers, Joe Bugner, a rematch with Joe Bugner, Jurgen Blin, Jean-Pierre Coopman. He turned a normally brutal profession into a pilgrimage.
Floyd Mayweather does not travel that well.
He lives here, his gym is across the freeway from the MGM Grand, and he invites the world to come to his side and peel him grapes.
On Saturday night he effectively mocked his own grandiose promotion by performing on one parallel plane while 23-year-old Canelo Alvarez flailed in another.
Nice work if you can get it, especially when it isn't work.
Mayweather moved another step toward getting to his finish line undefeated with this blowout of Canelo, who would do well to move up to 160 pounds, where he will be challenged but not mocked.
"Chess and checkers," Mayweather explained it in the weeks leading up to this, and there is no question who the king was Saturday night, and who was the pawn.
In fact, it seemed that the theme of the show – "The One" – might refer to the number of rounds Canelo would get.
But, since this is Vegas, hold all tickets.
Judge C.J. Ross ruled this virtual shutout a … draw? Even the 116-112 and 117-111 verdicts on the other two cards seemed ludicrous.
Maybe boxing needs a good dose of voter suppression.
"I'm not in control of what the judges do," Mayweather said. "That's a strong, young fighter over there. My dad (trainer Floyd Mayweather Sr.) said I was a little tight, and he was right. If I had attacked earlier I might have gotten the stoppage, but we had a great game plan."
As usual, Mayweather respected his foe enough to get into frightfully good shape and concoct a plan. Instead of launching right hands, he went jabbing from the beginning, and Canelo cooperated by standing frozen for the first two rounds.
By the time Canelo decided to fight, Mayweather had found alleys for a right that, in terms of impact and accuracy, is still strangely underrated. By the middle of the fight he was toying with Canelo, getting himself stuck in corners to test himself, taking a couple of unnecessary shots, just to say he'd been in a fight.


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Cough-mixture thief arrested after allegedly breaking into three clinics
Published on Aug 29, 2014 9:58 PM
SINGAPORE - A 35-year-old man who is believed to have broken into three clinics along Bedok South Road, Marine Terrace and Bedok North Street 1 was arrested on Friday.
He allegedly stole more than 100 bottles of cough mixture together with other medication like Panadol, sleeping pills, pain killers and cash amounting to about $2,250 between Aug 23 and Aug 29, the police said on Friday.
A raid of the suspect's residence unearthed 66 bottles of cough mixture together with other medications. Preliminary investigations revealed that the suspect may have gained entry into the clinics through the rear window or the front door.


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Complex PTSD
Dr. Judith Herman developed the concept of Complex PTSD. She argues that this new diagnosis is necessary to describe the symptoms experienced by survivors of long-term trauma such as child sexual abuse, prostitution, and organized child exploitation rings.
The National Center for Post-Traumatic Stress Disorder has a very helpful Fact Sheet on Complex PTSD.


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Video: Tags: Mad Men
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TAVIS SMILEY | John Slattery | PBS (#2622)
The "Mad Men" star discusses his character being the 'quintessential' 1960s male and his misgivings on going out of his comfort zone. To see this entire interview when it becomes available, go to:... 03:53
Tavis Smiley | Writer-producer Matthew Weiner | PBS (#2564)
Full Program: The award-winning writer-producer discusses Mad Men and the show's future with AMC. 24:07
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Tavis Smiley, the first African American to have his own signature talk show on NPR, hosts a new late-night television talk show - a hybrid of news, issues and entertainment, featuring interviews with newsmakers, politicians,...


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Gay ban repealed, but restrictions remain
WASHINGTON While President Barack Obama this week is expected to clear the way for gays to serve openly in the military, the new law won't go into effect immediately and unanswered questions remain: How soon will the new policy be implemented, will it be accepted by the troops and could it hamper the military in Afghanistan and Iraq?
The historic action by Congress repeals the requirement, known as "don't ask, don't tell," that for the last 17 years has allowed gays and lesbians to serve, but only if they kept quiet about their sexual orientation. Ending that policy has been compared in its social implications to President Harry S. Truman's 1948 executive order that brought racial equality to the military.
After Obama signs the legislation - passed by the Senate on Saturday - into law, the Pentagon must still certify to Congress that the change won't damage combat readiness.
So, for the time being the restrictions will remain on the books, though it's unclear how fully they will be enforced. Some people believe gay discharge cases will be dropped as soon as Obama signs the law. Military leaders, who have been divided on the issue, gave indications that the policy change will be aggressively pursued.
Marine Corps Commandant Gen. James Amos, who had argued against the policy change, said in a statement Sunday the Corps "will step out smartly to faithfully implement this new policy" and that he would "personally lead this effort, thus ensuring the respect and dignity due all Marines."
The issue of gays in the military has been a contentious one for decades. Until 1993, all recruits had to state on a questionnaire whether they were homosexual; if they said "yes," they could not join. More than 13,500 service members were dismissed under the law.
In the 17 years since the "don't ask, don't tell" policy went into effect, views toward the gays in the broader society have evolved. Gay marriage is now legal in five states and the District of Columbia. Opinion surveys say a majority of Americans think it's OK for gays to serve in uniform.
Peter Mansoor, a retired Army colonel who commanded a brigade in Iraq, said he believes the military - from top commanders to foot soldiers - will accept their new orders.
"Pretty much all the heated discussion is over and now it's a matter of the more mundane aspects of implementing the law," Mansoor, a professor of military history at Ohio State University, said in a telephone interview.
Aaron Belkin, director of the Palm Center, a research institute at the University of California, Santa Barbara said only three steps are needed to assure a smooth and quick transition: an executive order suspending all gay discharges, a few weeks to put new regulations in place, immediate certification to Congress that the new law will work. But he said the military may require months of education and training well into 2011.
Defense Secretary Robert Gates, who has supported the change but has stressed a go-slow approach, said "successful implementation (of the new policy) will depend upon strong leadership, a clear message and proactive education throughout the force."


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Versatile thyme adds its warm, aromatic flavor to slow cooked stews, soups, and roasted meats and poultry. Add it to stuffing, vegetables, beans and jambalaya.
Organic Thyme Leaves
Its sweet, minty-green flavor is sensational on poultry, fish, meats, vegetables, Cajun dishes and stews.
Serves: 4Prep time: 15 mins
This product has no significant nutritional value.


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Whale helping merchants reel in business
Bayfront visitors hope for a glimpse
Background: A Pacific gray whale has been sighted in San Diego Bay since March 10. It was migrating between Baja California and Alaska.
What's new: People on a tour boat saw the marine mammal, dubbed Diego, near Harbor Island yesterday morning.
What's next: There are no plans to help Diego leave the bay. Diego's health status is unclear, but gray whales can survive with little or no food during their weeks-long migration to Alaska.
News flitted through the San Diego Maritime Museum yesterday morning: Diego the stray whale had surfaced just in time for the 11 a.m. boat tour that highlights the bay's history and industry.
The timing was so perfect that Kathy Christenson, a tourist from Minnesota, feigned skepticism.
"Maybe it's a fake," she said, spinning a theory about Diego being a secret part of the federal government's economic stimulus plan.
What can't be disputed is that Diego – the young Pacific gray whale that adopted the bay three weeks ago today – has turned from a novelty into a moneymaker for a handful of bayfront proprietors, including eateries and boat-tour companies.
Diego often hangs out near the museum, prompting the organization to double its number of narrated bay tours aboard the boat Pilot.
Some of the businesses half-seriously suggest feeding the creature so it will provide regular entertainment as in a SeaWorld marine park.
"We are thinking about putting him on the payroll," said Mary Cobb, an assistant manager at Anthony's Fish Grotto on the Embarcadero.
She said that a few people call daily for Diego updates and that some stop for meals at the restaurant because the whale has been spotted from Anthony's dining room and patio.
"They are very curious to see him, and they enjoy our good food. They get a little of both," Cobb said.
She and others would like Diego to reside year-round, except they realize that it's time for the whale to rejoin the roughly 20,000 gray whales heading toward Alaska for the summer. If the whale waits much longer, it will miss the usual migration period.
Since March 10, the 30-foot whale has made several forays toward the bay's outlet and reportedly swam about two miles out to sea Wednesday, only to turn back.
Yesterday, an official with the National Marine Fisheries Service said the agency has no plans to entice Diego to leave the harbor.
Records show that at least one Pacific gray whale's detour has lasted longer than three weeks: Humphrey remained in San Francisco Bay and the Sacramento River for 26 days in 1985.
Before Diego arrived, the Pilot boat offered five tours per day Friday through Sunday. Since March 16, it leaves four to six times daily with mostly full loads of people. Diego is featured prominently on the museum's Web site, sdmaritime.com, along with tour details.
Yesterday, museum guide Mark Gallant prepared tourists for the 11 a.m. excursion. Then he joked about charging a special fee if Diego emerged.


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Proper puppy-proofing sets tone
Simple steps can make house safer and more enjoyable for young pets
• Tape loose electrical cords. Use outlet covers.
• Store cleaning chemicals out of reach. Use baby latches on cabinet doors if needed.
• Move breakables and valuables out of the puppy's reach. Roll up new or valuable rugs until the puppy is house-trained.
In your yard:
• Do not leave a puppy unsupervised, and be sure to fence in your yard before letting the puppy off leash.
• Use pet-safe gardening products, and be sure any lawn service you use does the same.
• Store grill utensils out of the puppy's reach.
• Make sure a swimming pool is fenced.
• Check the ASPCA's list of poisonous plants.
• As soon as puppies are old enough, start training them, especially the "drop it" and "leave it" commands.
Online resources
• The ASPCA's "Springtime Safety Tips," including a list of poisonous plants: www.aspca.org/pet-care/springtime-safety-tips
• For fun, see Design Sponge's "11 Ways to (Stylishly) Pet-Proof Your Home": www.designsponge.com/2014/01/10-ways-to-stylishly-pet-proof-your-home.html


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Hurricane Preparedness Center - Flint, MI
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Flint, Michigan
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Introduction to preparing for a hurricane
All Atlantic and Gulf of Mexico coastal areas are subject to hurricanes. A reliable estimate of expected annual property loss due to hurricanes is $5 billion. The Atlantic hurricane season lasts ...
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Metro Arts Alliance of Greater Des Moines
Rating: 5 stars 1 1 review 203
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Location: 500 E Locust Ste 201 Des Moines IA 50309 USA
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I have known Metro Arts Alliance as a member of the general public, a volunteer, and a board member for Metro Arts Alliance over the past 20+ years. They provide the unique service of linking our community to the arts through creative programs and alliances with community groups and other non-profits. Their approach has strengthen the arts and the creative economy in the Des Moines metro area for many years. MAA educational programs with the schools is second to none and has aided in providing a foundation of art appreciation for our children in this era of declining school funding. MAA constant search for new and high-value alliances and programs in our community make it a treasure among non-profits nationwide.
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He is a metaphor for the chicest Indian garment — the bandhgala. Over the years, he has reinvented the classic staple and recently even reinterpreted it for women. Dazzling bespoke junkies with his restrained and elegant aesthetic seasons after seasons, he has shied away from fashion weeks and branched into other ventures like hospitality and chocolates. The suave and elegant Raghavendra Rathore speaks about his design philosophy and plans...
You studied design overseas. How has the global perspective shaped your quintessential Indian aesthetics? What's the root of your artistic expression?
International exposure and worldly education is key to understand the value and the richness of our culture. In the global modern village, we need to comprehend and value the importance of 'indigenous design'.
You are one of few Indian designers who focused early on brand expansion — in terms of hotel properties, chocolatier etc...
The design landscape in India is unique in every regard. All my early experiences of working in the US made me stress on the fact that we wanted to make a design company and not only a fashion company. Hence the idea of diversity was key in the early years of building a diverse foundation or the Raghavendra Rathore label.
Of late you have stayed away from runway and fashion weeks (the bridal week was a one-off showcase in Mumbai). Any particular reason?
I believe the bespoke segment that we are in, may perhaps not need so much cajoling as the other segments of fashion. Flirting with the fashion weeks is important but very sparingly, time to time, depending on what the vision of the company is, we are open to such platforms.
You've redefined the classic bandhgala. Can you elaborate on the classic love affair...
Resurrecting the classic clothing from our heritage is not only important but crucial for the future generations. Modernising ideas from the past for the present is where we find our challenges for the company — it is why we exist.
What are your views on menswear in India? Why is it still in the nascent stages?
As retail gets more organised we will see the real value of Indian men's wear reach its optimum. The present speed at which it is growing is rapid and alarming, capturing the mindset is a real challenge for a design house like us.
Your design process and muse? What's in store for your next collection?
In spite of fashion being cyclic, each season we focus on deeper roots of heritage to bring out value from the old world and translate that in contemporary methodologies. We have varied interests in design and lifestyle which covers the entire bandwidth of lifestyle.
Any new ventures to reach out to the masses or are you looking at any brand tie-ups.
Capitalising over the hand-made Jodhpuri bandgala jacket, Jodhpuri pants, Jodhpuri achkans and other classic outfits is our present focus for the year. Providing an access to our services pan India will be the key area of growth over this and the coming year.


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Why oh why did Kosakowski go pro?
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Originally Posted by
naw. dk left the team on good terms with the coach and teammates. last year, he had been playing all of the usa futures in between his college matches, skipping most of the fall tournaments including last year's ITA All-Americans. at ojai, even though most top pac 10 players play, typically ucla's top players (kohloffel, abid, srugo) in years past do not compete, often playing only in the doubles draw or not at all. basically it was always dk's plan to go pro and thats why he played all of the american futures while in college. the question was when he should move on. and for dk, he is really dedicated to tennis and he sees tennis as something he wants to do for a long time. for those talking about if dk will need to pay for tution if he goes back to school, i dont see dk going back to college either. he left cause he wants tennis to be his career.
Fair enough. But realize that for 98% of players on tour, its a money-losing proposition. It costs a lot to compete year after year. And then you come back and that scholarship is gone.
I dunno, an education worth, lets call it $100K, fine, is still worth a lot. UCLA room, board, books etc is pretty pricey.
Degree would come in handy when trying to get a real job in a tough job market when tennis doesn't work out.
Trying to go back to college at 28-32 years old and pay full boat...well, good luck with that, most don't do it. He'll end up a country club pro at best. Nothing wrong with that...I guess.
You're telling me he couldn't compete in 3 more years? Didn't hurt Isner..at all. Young could have benefited from it too.
Anyway, it's his life. I get it. But its not now or never. Let him grow physically, learn the game more, play top collegiate players. Just seems shortsighted.
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Jonas Karlsson was born on March 11, 1971 in Salem, Stockholms län, Sweden as Sven Bert Jonas Karlsson. He is an actor and writer, known for Miffo (2003), Offside (2006) and Storm (2005). See full bio »
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The 5 Keys to setting up your successful garden.
Put your effort into building good soil for easier garden maintenance and long lasting
Step by step instructions for getting your crops growing.
Lactuca sativa
The brilliant green, oak-shaped leaves of this early, compact variety form a tight, compact rosette with a soft buttery texture, crispy ribs and sweet flavor for salads and sandwiches. Succession plant at close spacing in spring and fall to keep a constant supply.
Days to Harvest: 55
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NEWORLEANS- Landry-Walker High School in Algiers is mourning the loss of one of its brightest stars.
School officials say 17-year-old Johan Kenner was a personable, loving kid and leader on both the football and basketball teams. Kenner, known by friends and family as 'Yogi' died in a hail of gunfire Sunday evening in St. Roch Park.
'He just was a real good kid,' said his mother Cortina Johnson-Cash. 'All he wanted to do was graduate and go to college and play whatever because he played football, basketball and baseball. He was really, really good at all of them.'
Johan's mother says her son just had ACL surgery. 'Everyone started running and unfortunately, Johan couldn't run as fast because of his ACL surgery.'
Neighbors say the shooting makes little sense. 'For someone to shoot up a park, I mean this is a beautiful park,' said Abdi Farah from the St. Roch Community Church. 'It's kind of a center of the neighborhood.
Usually it's full of kids. Violence of any kind is just unnecessary, but to be around so many innocent people, it's kind of a loss for words.'
Johan's mother says she's not sure who the intended target of the shooting might have been. But, she says it appears the shooter just sprayed St. Roch Park with bullets. 'He just came in the park and started shooting toward the basketball goal and started shooting towards the playground area for the kids and then all over,' said Johnson-Cash. 'People just trying to live and to be going to a park, where kids play, a lot of kids, that just wasn't right.'
Johan was the second of nine children in his family. His mother is a correctional officer at Orleans Parish Prison. She now wants to move out of her house, just down the street from St. Roch Park, for the sake of her kids.
'I need to move out,' she said. 'New Orleans is just too dangerous to even really raise your kids here. I don't know. It's really dangerous.'
'This neighborhood has so much good, so many good families, so many good people,' said Farah. It's just weird how these little bursts of violence just keep coming up.' New Orleans police released few details about the shooting and said so far they have no motive or suspects in the case.


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DLC Character for Injustice Gods Among Us That I want to see
This will be updated if any of these characters make it as playable characters as DLC . There will be more characters on this list than probably in the game. The characters that make it will be removed from this list. These are the 20 I want to see, this list will change names once all DLC content is finished. The list will probably change to characters I wish were in Injustice


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Novell: No SUSE Linux for ARM-based netbooks
A year ago, netbooks were viewed as a good chance for Linux to finally break into the desktop computing mainstream.
A year ago, netbooks were viewed as a good chance for Linux to finally break into the desktop computing mainstream.
Advanced Micro Devices agreed to an Intel offer to make public the terms of a cross-license agreement at the heart of a dispute between the two companies. But there's one condition: in return, AMD wants Intel to lift a demand that evidence submitted ...
Lenovo has decided to unveil photos of its Pocket Yoga concept PC, an ultra-compact portable that's slim enough to slide in a back pocket. The leather-bound device, which resembles an oversized wallet, features a QWERTY keyboard and a touch screen.
Nokia is seriously contemplating entering the notebook market, according to research firm DisplaySearch. A blog entry by analyst John Jacobs quotes Nokia CEO Olli-Pekka Kallasvuo as saying in an interview with Finnish national broadcaster YLE that th...
Microsoft Corp. will deliver a release candidate of Windows 7 to the public in late May, according to a report from a noted Windows Web site.
Hewlett-Packard will begin selling on Monday laptop batteries that are guaranteed to last three years.
Asustek Computer, which pioneered the netbook market with its Eee PC line, plans to further slim down its business units and reduce the variety of netbooks it sells.
We're already a quarter of the way into 2009 and the gadgets are coming thick and fast despite the recession that is gripping many countries.
New laptop and desktop designs are on tap as PC and hardware makers start tweaking components to take advantage of improved features in Microsoft's upcoming Windows 7 OS.
Scientists at the Massachusetts Institute of Technology (MIT) have developed a way to charge lithium ion batteries in seconds, instead of hours, that could open the door to smaller, faster-charging batteries for cell phones and other devices.
One Laptop Per Child is set to dump x86 processors, instead opting to put low-power Arm-based processors in its next-generation XO-2 laptop with the aim of improving battery life.
Microsoft patched the first critical vulnerability in Windows 7 Tuesday as it rolled out an update that fixes three flaws in the new operating system's kernel.
Dell on Tuesday is expected to launch the tough Latitude E6400 XFR laptop with a touch screen, which the company claims can withstand drops, dust and high-pressure water sprays.
Rumors of an Apple netbook gained momentum Tuesday as the Dow Jones financial news service confirmed earlier reports that the company will launch a touch-screen-based computer, perhaps as early as the second half of this year.
Microsoft next month plans to release a toolkit to help business customers begin testing their existing applications for compatibility with Windows 7.
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First impression on unpacking the Q702 test unit was the solid feel and clean, minimalist styling.
For work use, Microsoft Word and Excel programs pre-installed on the device are adequate for preparing short documents.
The Fujitsu LifeBook UH574 allowed for great mobility without being obnoxiously heavy or clunky. Its twelve hours of battery life did not disappoint.
The screen was particularly good. It is bright and visible from most angles, however heat is an issue, particularly around the Windows button on the front, and on the back where the battery housing is located.
My first impression after unboxing the Q702 is that it is a nice looking unit. Styling is somewhat minimalist but very effective. The tablet part, once detached, has a nice weight, and no buttons or switches are located in awkward or intrusive positions.


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New tax rates in T-M, Calallen, Robstown
CORPUS CHRISTI - Calallen, Robstown and Tuloso-Midway independent school districts have Nueces County taxpayers to thank for the chance to gain more state funding for programs, new band equipment and teacher raises.
"I feel good," Calallen ISD Superintendent Artie Almendarez said after seeing the results for his district.
He, Robstown ISD Superintendent Alfonso Obregon and Tuloso-Midway ISD Interim Superintendent Sue Nelson said the opportunity for more funding wouldn't have been possible without voters' help.
"It's good for all three school districts," Nelson said.
Elections in each district asked voters to shift taxes from debt service to general operations.
Voters were asked to increase the maintenance and operations taxes to 1.17 per $100 valuation, then a state law will kick in allowing those districts to receive additional state funding based on a district's student attendance, with an emphasis on low-income students.
School districts must call for elections to increase their maintenance and operations tax rates to more than 1.04 cents, school district officials said.
Here are the final votes in each district and what they mean:
n In Calallen the tax rate change was approved 318 to 60.
Voters approved a 13-cent operation tax rate increase. Since that was approved, trustees plan to reduce the debt service tax rate by 13 cents. That would keep the overall tax rate the same. Calallen may receive an estimated $700,000 annually from the state thanks to the voter-approved change.
n In Robstown the change was approved 410 to 23.
Voters approved adding 13 cents to the operations tax rate and adding an almost 15-cent increase to the debt service tax rate. Trustees plan to reduce the debt service tax rate by 13 cents. Robstown may receive about $2 million annually thanks to the change.
n In Tuloso-Midway the change was approved 110 to 7.
Voters approved adding 13 cents to the operations tax rate and cutting a corresponding 13 cents from the debt service tax rate. That would keep the overall tax rate the same. Tuloso-Midway may receive more than $1.9 million annually thanks to the change.
Officials emphasized that the main purpose of the elections was to gain more state funding, particularly at a time when state government officials expect an $18 billion budget shortfall.
Several districts across the state already receive this funding, including the Corpus Christi Independent School District. Orange Grove Independent School District taxpayers recently approved a similar tax rate adjustment.
Calallen, Tuloso-Midway and Robstown officials have said they plan to put some of those extra funds toward teacher raises or assistance with offsetting increased health care costs.
In Tuloso-Midway, the money generated generally would go toward debt service, but this year it could go toward one-time expenses such as parking lot repairs, middle school and high school band instruments and restorations to the high school track, Nelson has said.
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Interesting Sailboats
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New Ovni Evolution 43
Not sure that this new yacht has been discussed yet ? Has been talked about by Ovni within the industry for a couple of months now and has just been listed on their website but no other details there yet. Looks like their version of the Sense 43 - maybe Ovni's time has come now that chines have gone mainstream !
cheers Robert
Now, this is really interesting, thanks for posting.
Almost everybody knows that OVNI are great voyage boats but their design was getting old and they, besides being good boats, were not interesting boats in what regards design.
It has changed know. First with the evolution 52 and know with the 43.
(If you have more, or bigger images of the 43, please post them)
I am sure these boats will have a much better performance downwind and even downwind because finally alubat is installing naca profiles on both the board and the twin rudders. They talk about this to be a kind of Cigale centerboarder and that are just good news.
Quite a revolution going on OVNI, well they call it evolution
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text: Publications Search Results Users may wish to consult What's Included in the Bibliography. Results 1 - 2 Results per page: 25 50 100 Sort by: Pub Date (newest) Title Recent Additions Author Clear All Pub. Year Citation 1986 Grimes, D.A. . Declining surgical case-load of the obstetrician-gynecologist. Obstetrics and Gynecology. 67, (6), 760-762. Full Text Options: DOI Worldcat Google Scholar Export Options: RIS EndNote XML 1997 Learman, L.A., Grimes, D.A. . Rapid hospital discharge following laparoscopy for ectopic pregnancy. A promise unfulfilled?. Western Journal of Medicine. 167, (3), 145-148. Full Text Options: Worldcat Google Scholar Export Options: RIS EndNote XML [1]


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Search Results
Created by Dr. Naveed Ahmad, this website brings together hundreds of high-quality resources for people interested in learning more about the world of radiology. On the homepage, visitors are presented with a list of...
The Learning Radiology website was conceived and created by Professor William Herring of the Albert Einstein Medical Center in Philadelphia. The site was started in June 2002, and now receives upwards of 12,000 visitors...
These online radiology images are part of the MedPix database, which is offered as a public service by the departments of radiology and biomedical informatics at the Uniformed Services University in Bethesda, Maryland....
A unit designed to give students knowledge in the area of nuclear medicine as well as improve math skills. Students become familiar with nuclear medicine through diagnostic imaging and its components.
The Medical College of Wisconsin?s Collaborative Hypertext of Radiology (CHORUS) website contains over one thousand documents and images related to anatomy and physiology, radiological findings, diagnostics, and d...
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text:
by GM Ross - Steve Claggett, the fighting pride of Western Canada, is eager to get one of Ontario's lightweight prospects in an Alberta ring in the near future. According to Claggett's manager Eric De Guzman, his fighter is ready and willing to meet either Buzz Grant or Logan McGuinness in an eight round contest at 135lbs. Teofista Promotions has a card in the works for July 24th, to be staged in either Lethbridge or Calgary, for which a Claggett-Grant or Claggett-McGuinness bout would serve as the headliner..
All three fighters Claggett, Grant and McGuinness are red hot at the moment, representing spots two through four in the CBF rankings. Most recently, Buzz Grant thoroughly outpointed Michael Kelly of Ireland at the Royal York Hotel in Toronto, Ontario. In March, Logan McGuinness turned more than a few heads at the Montreal Casino with his unanimous decision victory over experienced Mexican journeyman Pedro Navarrete. The same evening McGuinness was outpointing Navarrete in Montreal, Steve Claggett was busy in Calgary, scoring a second round TKO over former Nova Scotia amateur champion Walter Linthorne.
Regardless of which two fighters come together, the bout will certainly have major implications for the CPBC and CBF titles. The winner will surely be considered the most legitimate opponent for number one ranked CBF contender Tony Luis and/or his CPBC counterpart Pier Olivier Cote, setting up a championship showdown for later in the year.
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text:
Air guitarists mock you like a hurricane
Originally Published: July 27, 2010By Patrick Hruby | Page 2
Think you're a pretty good air guitarist? A Slash of invisible ax-wielding? A maestro of the shred-like-no-one's-watching dark arts?
Trust me: You're a rank amateur. A dilettante. No matter how much of Van Halen's "Eruption" you've committed to fingertip muscle memory, you have nothing on the nation's top pretend guitarists.
And yes, our nation actually has a cadre of elite pretend guitarists.
Last week, the best and brightest gathered in New York ...
Read full story
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Millington board mulls operations for new library
Panel ponders management, site
As city officials ponder a location for a new library, the Millington Library Board is re-evaluating who will handle its day-to-day operations.
The library board will decide in the next six months whether to renew its contract with the Maryland-based Library Systems and Services or go a new direction.
LSSI has one year left on its three-year contract worth $476,000 to provide privatized library management and staffing services, but the city must give notice on Nov. 1 whether it will renew the contract.
"The (library) board, in cooperation with the mayor, has decided to re-evaluate all options regarding the best way for the library to go forward," said Brian Miller, director of the Millington library.
Although Miller said library officials have been happy with LSSI's management, Millington has the option of either running the library itself or using the Memphis and Shelby County library system.
If the city does decide not to renew the LSSI contract, Miller said the Millington Library Board is leaning strongly toward having the city manage the library.
"I am confident that the city of Millington has the ability to oversee the operation of the library directly," he said. "The board has shared the same feeling, emphasizing their own confidence with the current staff of the library as it now stands."
The last option is to go back to having Memphis and Shelby County run the library, but library officials said recent staffing shakeups in Memphis make that option less appealing.
The library was a branch of the Memphis and Shelby County system five years ago, but when the county stopped funding the branch, Millington split from the larger system and hired LSSI.
"Most folks can make their own judgment on the headlines in the papers about neighborhood library closings and changes in management dictated by political decisions," Miller said. "That, in my honest opinion, is not the best way to maintain a stable, productive and enduring library."
Regardless of what direction the library board decides to take, choosing where to relocate the facility will be one of the main goals for library leaders.
Millington has nine years to find or build a new library before it turns over the current Navy Road building to the Millington Telephone Co. as part of a trade for USA Stadium.
City Mayor Richard Hodges said the library board and city officials are reaching out to residents to provide feedback on locations for the new library.
One local landowner has already offered a lot off of Easley Street, but city leaders are open to any ideas.
"We would love for people to bring us comments and suggestions," he said. "This is a major decision and we have to decide where we want to go."
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- Exchange: EN Paris
- Sector: Technology
- Industry: Hardware
- Sub-Industry: Consumer Electronics
Archos+ Add to Watchlist
As of 07:14:34 ET on 05/28/2015.
Company Profile for Archos (JXR)
Archos manufactures computer peripherals and digital entertainment products. The Company produces the Jukebox line of products that combine MP3 portable recorders and players, photo albums, digital cameras, digital camcorders, and video recorders and players. Archos also manufactures portable hard drives, CD recordable and rewritable devices, and DVD players.
Contact Information
Key Executives for Archos (JXR)
|Henri CrohasChairman/CEO/Founder
|Loic PoirierChief Operating Officer
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WILSONVILLE, Ore. An armed man tied up two Wilsonville La Quinta Inn employees and robbed them and the hotel Friday morning, officials said. Police were investigating whether the robbery was linked to a similar hotel robbery on Thursday.
The suspect was wearing a ski mask when he entered the La Quinta Inn at 8815 SW Sun Place around 4:15 a.m. Friday, according to Deputy Bryon O'Neil of the Clackamas County Sheriff s Office.
Two women were preparing breakfast when the suspect pointed a gun at them and tied them up, O Neil said. He then un-tied one woman and made her empty a cash drawer.
The women said the robber also took both of their purses before fleeing the scene. No one was hurt during the robbery.
The suspect was described as a man in his early 20s of medium height and weight, wearing a long shirt, gloves and ski mask.
The Portland Police Bureau was investigating whether the robbery was connected with a similar incident that took place Thursday at the Silver Cloud Inn in Portland. Anyone with information about either robbery was asked to contact the police.
Related: Suspect ties up, robs Portland hotel employee


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In This Section
AES News
- First Book in "AES Presents" Series from Focal Press
- New edition of Handbook for Sound Engineers, edited by Glen Ballou
- 137th Audio Engineering Society Convention Breaks Records and Draws Acclaim from Attendees, Exhibitors and Presenters Alike
- Convention reminds West-Coast audio community, "If It's About Audio, It's At AES!"
- AES 2014 Election Results
- The results are in!
- Time to Vote: 2014 AES Elections
- Deadline was Friday, July 11th
AES Silver Medal Award
Tomlinson Holman
The AES recognizes and honors those who have made outstanding contributions to the field of audio in engineering, technology, service, and the arts through its Awards Program.
In 2006, Tomlinson Holman was presented with the AES Silver Medal Award for more than three decades of work in the advancement of audio electronics, multichannel sound techniques, and motion picture sound production and reproduction standards.


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Mon-Fri: 7am-1am EST / Sat-Sun: 7am-11pm EST
ApplianceJenn-Air Range
Level of DifficultyReally easy
Time to do repair15 - 30 mins
Age of ApplianceMore than 10 years
NutdriverScrew drivers
10 out of 12 people found this instruction helpful
shut off power , remove vent cover, 2 screws to free the switch box , there was enough slack to turn it over , idendtify switches to be replaced , double check you got the right ones , got em , nut driver for the fan switch, torex for the burner switch , test ,put it back together , and clean up , no more than a half hour . order parts on monday , delivered on tuesday afternoon with 3-5 day shipping. thanks guys
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text:
This Year is continuing with its best-ofs of 1UP podcasts as we cover (for the first time) Retronauts. Retronauts was, of course, 1UP's great retro games podcast. It was one of the first podcasts I ever listened to, and maybe it's just podcast nostalgia (though I doubt it), but I'm convinced that it's one of the best gaming podcasts ever created. And it has been, of course, recently rebooted; I believe the new show kicks off next month. This is a best-of compilation only for the 2007 year of the show. We've done these compilation shows in the past for other 1UP programs, namely 1UP Yours and GFW Radio.
Within we hear discussed such topics as the history of survival horror, 8-bit Mario games, the Atari 2600, and the Commodore 64, among a number of other things. The best-of includes such luminaries as Jeremy Parish, Chris Kohler, Scott Sharkey, Shane Bettenhausen, and Jenn Frank. Here's a blurb I wrote:
Retronauts was part of the 1UP Network's expansion into podcasting in 2006, appearing alongside popular shows like GFW Radio and 1UP Yours. As dreary as host Jeremy Parish liked to suggest it might be—he termed it 1UP's only podcast "for old people"—the podcast was anything but. Though it might not have been as bombastic as its sister programs, Retronauts was in fact just as entertaining, thought-provoking and, more often than not, funny. A well researched, informative journey through video game history, Retronauts was a podcast of inestimable value: not only a rare thing—a history podcast that never bored—but an important aural vade mecum of the games so many people love today.
My favorite topic in the whole show is the survival horror one. It's a must-listen. You can download the best-of here (410 mins/188MB) and there's more information at our site. There you'll find a link to our iTunes show page, RSS feed, Twitter, et cetera. Thanks for your continued support! We receive a ton of feedback here which is great and is really appreciated. Please enjoy the program.


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Existential psychology
Existential psychology
a theory of psychology, based on the philosophies of phenomenology and existentialism, which holds that the proper study of psychology is an individuals experience of the sequence, spatiality, and organization of his or her existence in the world.
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text:
Economy's Tumble Even Worse Than Expected In 3Q
Posted: 11/25/2008 - The government says the economy took a tumble in the summer that was worse than first thought as American consumers throttled back their spending by the most in 28 years.
Posted: 11/25/2008 - The government says the economy took a tumble in the summer that was worse than first thought as American consumers throttled back their spending by the most in 28 years.
Posted: 11/25/2008 - The Colorado State Fair is going to a four-day work week to cut utility costs.
Updated: 11/26/2008 - A jet-pack pilot has flown across the 1,500-foot-wide, 1,100-foot-deep royal gorge in southern Colorado.
Posted: 11/25/2008 - The government has introduced a pair of new programs that will provide $800 billion to help unfreeze the market for consumer debt which Treasury Secretary Henry Paulson calls vital to supporting the economy.
Updated: 11/24/2008 - The Dow Jones surged 396 points Monday upon news that the federal government is set to bailout banking giant Citigroup. But while Wall Street celebrates, Main Street grows weary as more of their dollars are handed over to corporations.
Updated: 11/24/2008 - Mesa County has approved a plan to spend up to $1.5 million dollars on healthcare for inmates next year.
Posted: 11/24/2008 - Monty Python's Spamalot" will now close Jan. 11. A week earlier than the musical announced last month.
Updated: 11/24/2008 - With a bailout plan for automakers on hold in Washington, Grand junction car dealers are trying to clear their lots before the end of the year. Local car dealers say sales for their used cars are up but they say it may be a better investment to buy new.
Updated: 11/25/2008 - The Montrose County District Attorney's Office has filed formal charges against the teen suspected of stabbing a Montrose High School student.
Updated: 11/25/2008 - Turkey Day is fast approaching and one local woman says she would rather have the bird in her yard than on her table.
Updated: 11/24/2008 - Local area ski resorts receive grades on their efforts to go green.
Posted: 11/24/2008 - "Young Frankenstein" is closing after nearly 500 Broadway performances.
Posted: 11/24/2008 - Bruce Springsteen's new single can be downloaded free of charge for one day.
Posted: 11/24/2008 - Celebrity buzz about the AMA's and more
Posted: 11/24/2008 - Chris Brown has plenty to sing about.
Posted: 11/24/2008 - A state-run newspaper in China is blasting the latest Guns N' Roses album as an attack on the country.
Posted: 11/24/2008 - How would Obama's appointees work together?
Posted: 11/24/2008 - Authorities in Cincinnati say three children have died in a house fire.
Posted: 11/24/2008 - A constitutional showdown over presidential power could play out this morning in a Washington courtroom.
Posted: 11/24/2008 - President Bush is hosting Israeli Prime Minister Ehud Olmert for a private dinner.
Posted: 11/24/2008 - Iranian state radio says an alleged espionage network with links to Israel's Mossad spy agency has been dismantled.
Posted: 11/24/2008 - Mark Williams from Our Country Deserves Better talks about Palin Thank You ads
Posted: 11/24/2008 - Astronauts aboard the International Space Station think they've made some progress with a faulty water recycling system.
Posted: 11/24/2008 - More than 20 people have been killed in a series of bomb attacks today in Iraq.
Posted: 11/24/2008 - American auto companies are making a big push for the Chinese markets


text:
The Supreme Court Bar Association has opposed the Chief Justice of India R.M. Lodha's suggestion to Chief Justices of all High Courts that courts should function all 365 days in a year. The CJI had said "the courts should function all year round, giving individual judges the choice of holidays and vacations."
Responding to the CJI's suggestion, SCBA president P.H. Parekh said "on the face of it, 365 days work scheme may not reduce the arrears because if working hours of the judges remain the same the judicial output will remain constant."
Mr. Parekh, in a letter to the CJI, said, "In fact the new scheme may perhaps result in an increase in the arrears because on Saturdays, Sundays and other holidays, the stake holders in the administration of justice may not take the judicial work as seriously as they do it on working days."
He added, "It is believed worldwide that holidays are very necessary for the efficient and effective functioning of any institution. It will apply with greater force to members of the bar looking at their working hours. The view of most of the members of the SCBA is that they cannot possibly work for 365 days as no human being can or should work 365 days."
The Bar Council of India, after a meeting of State bar councils, had already voiced its opposition to the 365 days work scheme.
Mr. Parekh said, "The members of the bar are already working very hard under the existing system. Except for their work obligations, they have their family as well as social obligations. They need holidays to prepare their cases and they also need some time to have vacation as all others have."
Instead of the 365 days work scheme, the SCBA suggested among other things that vacancies of judges in the Supreme Court, High Courts and subordinate judiciary should be filled up expeditiously, unnecessary adjournments of cases should be avoided and, if necessary, working hours of all courts could be increased by one hour on all working days for some time as an experiment.
Keywords: Bar association, Supreme Court, court holiday, Justice Lodha, court vacation