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US 5792606 A Abstract The application describes homogeneous assays for determining ligands. A sample is contacted with a labeled substance P2 and receptor R. The receptor R consists of one molecule of P1, which monovalently and specifically binds to P2, and one molecule for R1, which binds to the ligand of interest. Complexes of P1, P2, R1 and ligand form, and are determined. R1 must be a nucleic acid molecule.
Claims(8) 1. Method for determining a ligand via a homogeneous assay, comprising:
contacting a sample with an amount of a labelled substance P2, and a receptor R which consists of one molecule of a binding partner P1, which monovalently and specifically binds to P2, and one molecule of R1, which specifically binds to said ligand, to form complexes comprising P1, P2, R1 and said ligand, and determining said complexes as a determination of said ligand in said sample, wherein R1 is a nucleic acid molecule. 2. The method of claim 1, wherein said homogeneous assay is a competitive assay and further comprises adding to said sample a conjugate C which consists of (a) a member of the group consisting of a molecule of ligand and a molecule of ligand analogue and (b) a molecule of P1, wherein said conjugate C competes with any ligand in said sample for binding to R1.
3. The method of claim 1, wherein P1 is biotin, a hapten, or an epitope.
4. The method of claim 1, wherein P2 is a member selected from the group consisting of avidin, streptavidin, a streptavidin polymer, an antibody, an Fab antibody fragment, and an F(ab')2 antibody fragment.
5. Method for determining a ligand in a heterogenous assay, comprising:
(i) incubating a liquid sample with two receptors R' and R" and an unlabeled substance P2 immobilized on a solid phase, wherein said R' consists of one molecule P1 ' and one molecule R1 ', and said R" consists of one molecule P1 " and one molecule R1 ", wherein P1 ' specifically and monovalently binds said P2, each of R1 ' and R1 " specifically bind to said ligand, to immobilize said ligand on said solid phase to form a complex comprising P2 -P1 '-R1 '-ligand --R1 "-P2 "--, (ii) contacting said immobilized complex with labeled P2, wherein P2 " specifically and monovalently binds labeled P2 to form a complex comprising P2 -P1 '-ligand --R1 "-P2 "-P2, and (iii) measuring a level of labeled P2 bound to said labeled phase as a determination of said ligand in said sample, wherein R1 ' and R1 " are different nucleic acid molecules. 6. The method of claim 5, wherein said heterogeneous assay is a competitive assay and further comprises adding to said sample a conjugate C which consists of (a) a member of the group consisting of a molecule of ligand and a molecule of ligand analogue and (b) a molecule of P1, wherein said conjugate C competes with any ligand in said sample for binding to R1.
7. The method of claim 5, wherein P1 is biotin, a hapten, or an epitope.
8. The method of claim 5, wherein P2 is a member selected from the group consisting of avidin, streptavidin, a streptavidin polymer, an antibody, an Fab antibody fragment, and an F(ab')2 antibody fragment.
Description This application is a continuation-in-part of application Ser. No. 08/887,587, filed May 20, 1992, now U.S. Pat. No. 5,437,981, which was a continuation of application Ser. No. 07/656,393, filed Feb. 21, 1991, abandoned. The invention concerns an immunological method for the determination of a ligand as well as a suitable reagent composition therefor. Very many substances occur in body fluids and tissues which are capable of binding to a specific binding partner and serve as parameters for particular diseases or for the state of health of the human body. These include, on the one hand, immunologically active proteins which have binding sites on their surface such as e.g. tumour markers, hormones or viral proteins and, on the other hand, DNA fragments. Since these substances, which are denoted "ligand" in the following, often only occur in very small amounts, they are detected using methods which are based on immunoassays by which these substances can be determined very specifically and exactly. The known immunological methods of determination may be classified into homogeneous and heterogeneous procedures. In the heterogeneous procedures a solid phase reaction is always involved in order to immobilize complexes containing the substance to be detected and a labelled constituent and thus to separate them from unbound constituents. In homogeneous methods no separation of bound label and unbound label is carried out so that differentiation between bound and unbound label must be effected by other methods. The heterogeneous immunoassays are mainly based on two variants namely competitive and sandwich assays. In these variants of the method at least two receptors are usually employed which are capable of binding to the ligand to be detected and of which one carries a label and the other is bound to a solid phase or mediates the binding to the solid phase. A multitude of variants for this are known in which further receptors are used. All complexes which are bound to the solid phase and which are labelled then enter into the determination. Basically there are two variants for carrying out competitive heterogeneous immunoassays in which either an antibody to the ligand is immobilized or a substance analogous to the ligand is immobilized. In the first variant a sample solution containing the ligand and a conjugate of ligand and a label is incubated with the immobilized antibody. In this process ligand and labelled substance compete for the binding to the antibody. The more ligand is present in the solution the less labelled substance can be bound. After separating the solid and liquid phase the label can then be determined in one of the two phases. The amount of bound labelled substance is an indirect measure for the amount of substance to be determined and thus of ligand. In the second variant the sample solution containing the ligand is incubated with an antibody which is specific for it and with the immobilized analogue of the substance. In this process the immobilized ligand and the ligand present in the solution compete for binding to the antibody. The more ligand is present in the solution the less antibody is bound to the solid phase via binding to the immobilized ligand analogue. Also in this case, after separating the solid phase from the liquid phase, the amount of bound label is determined which again is indirectly proportional to the amount of ligand in the sample solution. In the variants of the sandwich immunoassay an antibody against the ligand is immobilized and the sample solution is incubated in the presence of this immobilized antibody together with a further antibody which is capable of binding to the ligand and is labelled. In this process the receptors are added in excess so that all molecules present in the sample which are to be detected are bound to the solid phase and bind to a labelled receptor. A disadvantage of this method is that for each substance (ligand) to be determined several specially adapted receptors have to be provided. Thus, labelled special antibodies are often necessary which are different from ligand to ligand. The covalent binding of labelling groups to antibodies, which usually attack at several sites on the antibody molecule, can result in undesirable changes in the binding properties of the thus modified antibody. A further problem occurs when testing for proteins or DNA or RNA fragments which have more than one binding site. In these cases a majority or a multitude of receptors can bind whereby the resulting inaccuracy may still be compensated by using standards. This inaccuracy is, however, potentiated when all the receptors used are themselves bivalent or polyvalent. It was therefore the object of the invention to provide a method with which ligands can be detected with high accuracy, good reproducibility and high specificity and in which, in addition, universally applicable labelled constituents and universally suitable solid phase materials can be used. In particular, only one single specially adapted receptor should be required. This object is achieved by a method for the immunological determination of a ligand which is characterized in that the ligand to be determined is reacted with a) at least 2 molecules of a specifically bindable substance P2 of which at least one of these molecules carries a label and b) a receptor R which consists of a binding partner P1 which is capable of monovalent binding to P2 and a binding site R1 for the ligand to be determined and then the label is determined. According to characteristic a) labelled P2 is used in a known amount and preferably in excess with respect to the possible amount of ligand whereby this excess relates to the double stoichiometric amount of ligand if all of P2 is labelled and to the single stoichiometric amount of ligand if unlabelled P2 is also used. In the latter embodiment unlabelled P2 is also present in a one-fold excess. The method according to the present invention is suitable for the determination of all ligands to be detected in body fluids or tissue extracts which are capable of a specific binding whereby substances in low concentrations can be detected just as well as those in high concentrations. The sensitivity and accuracy of the method is improved compared to the hitherto known methods. The invention allows rapid and reliable determinations to be carried out with simple reagents. The method is particularly suitable for determining ligands with several binding sites and DNA or RNA molecules. In this connection, a specific binding site is understood as a binding site which can participate in a specific binding to another substance. Examples of this are antigenic determinants, specific binding sites on proteins or a specific nucleic acid sequence on DNA or RNA. An important feature is that receptor R contains a single partner P1 of the specific binding pair P1/P2 and this partner P1 is monovalent with respect to P2 so that only a single molecule P2 can be bound per molecule of receptor R. Substances are suitable as component R1 which are capable of specific binding to the ligand. These can be e.g. macromolecules such as antibodies, antibody fragments and specifically bindable binding proteins, haptens, epitopes or, in the case of a DNA or RNA test, DNA probes. The component R1 preferably has only one binding site for the ligand. In the case of a protein test it is preferable to use antibody fragments which only have one paratope or substances which bind specifically to the protein, for example T4 for the detection of T4 binding protein. For the detection of DNA or RNA, a probe is for example used as component R1 which can hybridize with a sequence of the DNA or RNA to be detected. A Fab fragment of an antibody capable of specific binding to the ligand is especially preferred as the component R1 in the receptor R. Depending on the ligand, R1 can be a uniform substance or a mixture. The component P1 of the receptor R is a partner of a specific binding pair. Pairs which can specifically bind to one another are known. Suitable binding partners are especially biotin-streptavidin or avidin; hapten-antibody, antigen-antibody, concanavalin-antibody; sugar-lectin; hapten-binding protein, e.g. thyroxine binding globulin and thyroxine antibodies or oligopeptide antibodies. Biotin and streptavidin or avidin are particularly preferably used as the binding pair so that receptor R1 especially preferably contains a single molecule of biotin as partner P1. The production of the conjugates is carried out using methods known to the expert (e.g. in analogy to Eur. J. Biochem. 131 (1980) 333-338). In the case of a heterogeneous procedure P2, in a non-labelled form, mediates the binding to the solid phase. In accordance with the present invention, the provision of a universal test is achieved since solid phase and labelled conjugate can remain the same for all determinations and only a single receptor has to be matched to the respective ligand to be detected. Since the binding of R, ligand and labelled P2 takes place in a homogeneous phase it is preferred over the binding of R to the solid phase P2 which proceeds in a heterogeneous phase. The unlabelled P2 of the specific binding system which is bound to the solid phase is therefore used in excess. Conjugates are preferably used as receptor R which consist of an antibody or antibody fragment R1 and biotin as P1. The production of the conjugates R is carried out in a known manner by reaction of the component R1 e.g. of an antibody or antibody derivative, with the partner P1 in a stoichiometric 1:1 ratio. This coupling reaction preferably takes place via a free SH group or amino group. The products which form statistically in this process containing two or more components P1 can be separated by gel chromatographic methods. For the method according to the present invention conjugates R are particularly preferably used which consist of a Fab' fragment and biotin. In order to produce such conjugates an antibody capable of binding to the ligand to be detected is treated for example with pepsin, the F(ab')2 fragment formed is subjected to reducing conditions and is subsequently reacted with the component P1 whereby the component P1 binds to a free SH group or amino group of the Fab' fragment either via a functional group present in the component P1 or via a spacer, if desired after activation of the binding sites. Products which contain one or more partners P1 can be separated for example according to one of the procedures described in EP-A 0 131 343 in which the separation is based on the difference in charge of related proteins. If the partner binds via the amino groups of a protein then single or several-fold binding leads to conjugates of different charge which enables their separation. Particularly suitable as the solid phase are test tubes or microtitre plates made of polystyrene and similar plastics the inner surfaces of which are coated adsorptively or covalently with P2. Particulate substances are also suitable e.g. molecular sieve materials, glass beads, plastic tubing and similar materials as well as porous carriers in a layer form such as paper. The binding of the partner P2 is carried out in a known way. At least a portion of P2, in the homogeneous test procedure all of P2, is employed in a labelled form. The usual labels for immunological tests are suitable as the label e.g. an enzyme, a fluorescent, chemiluminescent or radioactive substance. Met hods for labelling are known to the expert e.g. from Clin. Chim. Acta 81 (1977) 1-40 and do not need to be described here in further detail. The label can be determined in a known way. If the invention is used with a homogeneous test procedure, i.e. without use of a solid phase, then all of P2 is used in a labelled form. The complexes which form in this process of ligand, at least two molecules R and a number of molecules of P2 corresponding to the number of R in the complex are labelled and then detected by methods known to the expert for measuring a label in homogeneous test procedures. Suitable methods of detection are described for example in EP-B 0084 807. When carrying out the method according to the present invention in a heterogeneous as well as in a homogeneous way a conjugate C can be used in addition which contains the ligand or a ligand analogue with a binding partner P1 which is identical to the binding partner P1 in the receptor R. In this procedure ligand in the sample solution competes with ligand or ligand analogue in the added conjugate C for the component R1 in the receptor R so that a competitive method results. This embodiment can be applied to solid phase methods as well as to methods in a homogeneous phase. The details of the production of the receptor R apply in the same way to the production of the conjugate C. In this connection a substance is regarded as the ligand analogue which has similar properties to the ligand itself with regard to capability of binding to component R1 in the receptor R so that under the stoichiometric conditions employed competition does indeed take place for the binding site. The method can be carried out in one or more steps. The evaluation is carried out in a known way. Since each of the receptors and also the substance to be determined can only react specifically with its own particular reaction partner it is possible to incubate all receptors and the sample together and to carry out the method in one step. This is particularly advantageous when carrying out the method in an automated analyser. If the method is to be carried out heterogeneously in one step then substance P2 which carries a label is preferably employed in a limiting amount compared to receptor R. All variants of the method are preferably carried out in a buffered solution. Buffer systems for this method are generally known. GOOD buffers and phosphate buffers are particularly suitable for this. In order to carry out the method according to the present invention in a heterogeneous phase the sample solution is incubated either simultaneously or sequentially with the three receptors R, labelled P2 and, if desired, C in the presence of a solid phase coated with unlabelled P2. In this process two receptors R bind for example to the ligand via the component R1. Labelled P2 is then bound via partner P1 of the one receptor R and binding to the solid phase occurs by means of the binding pair P1/unlabelled P2 via the other receptor R. All complexes which are bound to the solid phase and which carry a label enter into the evaluation. According to the present invention a method is provided which is simple and which can be carried out rapidly and which is very sensitive even when using polyclonal antibodies. The present invention also provides a reagent for the immunological determination of a ligand which is characterized in that it contains a specifically bindable substance P2 which is present at least partially in labelled form and a receptor R which consists of a binding partner P1 which is capable of monovalent binding to P2 and a binding site R1 for the ligand to be determined. The reagent preferably contains the substance P2 in a labelled form and in a form bound to a solid phase in a combined formulation (e.g. a layer in a test carrier device or magnetic particle solution). According to a preferred embodiment this reagent contains unlabelled P2 bound to a solid phase or capable of binding to a solid phase. According to a further preferred embodiment the reagent contains in addition a conjugate C of ligand or ligand analogue and a component P1 as defined above. This reagent is suitable for the determination of a multitude of parameters in body fluids and tissue extracts. In a preferred embodiment the reagent contains in addition buffer substances. It particularly preferably contains phosphate buffers or GOOD buffers. FIG. 1 shows a diagram in which the results for the determination of the thyroxine binding index are plotted. FIG. 2 shows a diagram in which the results for the determination of anti-T4 antibodies are plotted. The invention is elucidated by the figure and by the examples. Determination of AFP (α-foetal protein) a) Production of a conjugate of biotin and Fab' fragments of anti-AFP antibodies (anti-AFP-Fab'-biotin) Polyclonal antibodies against AFP are purified immunosorptively and Fab' fragments are produced therefrom. These are coupled to biotin according to Analyt. Biochem. 161 (1987) 262-271 or Analyt. Biochem. 149 (1985) 529-536. b) Test procedure Buffer A: 120 mmol/l sodium barbiturate 18.2 mmol/l phosphate buffer, pH 8.6 1.27 mmol/l 8-anilino-1-naphthalinesulfonic acid 0.2% by weight bovine serum albumin (final concentration in the test) 50 μl sample (human serum supplemented with AFP) is added to 480 μl buffer A and 20 μl anti-AFP-Fab'-biotin (final concentration in the test: 4 μg/ml) in a polystyrene vessel coated with streptavidin (produced according to EP-A 0 269 092) and incubated for 30 minutes at 25� C. Subsequently 480 μl buffer A and 20 μl of a solution of a streptavidin-POD conjugate (50 mU/test) is added and incubated for 30 minutes at 25� C. It is washed and 1 ml ABTS� solution (9.1 mmol/l ABTS�, 2,2'-azino-di 3-ethylbenzthiazoline sulfonic acid (6)!-di-ammonium salt, 100 mmol/l phosphate-citrate buffer, pH 4.4, 3.2 mmol/l sodium perborate) is added, incubated for 30 minutes at 25� C. and the optical density is determined at 422 nm as a measure for the AFP content. T-uptake test The determination is carried out in such a way that T4 is added to the sample in order to saturate excess TBG. The unbound T4 is then measured. A calibration curve is thus obtained which is directly proportional to the thyroxine binding index (TBI). Sample: standards containing defined amounts of TBG and T4 in human serum are used as standards. A TBI of 0.44 is obtained for sample 1 and a TBI of 1.44 for sample 2 (cf. instructions for the Enzymun Test� TBK of Boehringer Mannheim GmbH, order no. 249416). Reagent 1 100 U/l conjugate of streptavidin and POD 36 pmol/l T4 120 mmol/l sodium barbiturate 18.2 mmol/l phosphate buffer, pH 8.6 0.2% by weight bovine serum albumin Reagent 2 0.2 nmol/l conjugate of T4 and biotin 0.15 mg/l anti-T4 -Fab'-biotin of polyclonal antibodies against T4 (produced according to Example 1) 120 mmol/l sodium barbiturate 18.2 mmol/l phosphate buffer, pH 8.6 0.2% by weight bovine serum albumin 50 μl sample are added to 500 μl reagent 1 in a polystyrene vessel coated with streptavidin and incubated for 30 minutes at 25� C. 500 μl reagent 2 are added and incubated for 30 minutes at 25� C. Subsequently it is washed with water and 1 ml ABTS� solution (cf Example 1) is added, incubated for 30 minutes at 25� C. and the optical density is determined at 422 nm as a measure for the thyroxine binding index. The results are shown in FIG. 1. Determination of an antibody against T4 Standard solutions of polyclonal antibodies against T4 having the concentrations 0, 0.5 and 1.0 mg/l are used as sample. Reagent: 10 mmol/l T4 biotin conjugate 50 U/l streptavidin-POD conjugate 120 mmol/l sodium barbiturate 18.2 mmol/l phosphate buffer, pH 8.6 1.27 mmol/l 8-aniline-1-naphthalinesulfonic acid 0.2% by weight bovine serum albumin 50 μl sample and 1 ml reagent are incubated for 30 minutes at 25� C. in a polystyrene vessel coated with streptavidin. Subsequently it is washed and 1 ml ABTS� solution (cf Example 1) is added, incubated for 30 minutes at 25� C. and the optical density is determined at 422 nm as a measure for the content of antibodies against T4. The results are shown in FIG. 2. Preparation of IgG biotin (1:1) 1) 50 mg of a monoclonal antibody against TSH (ECACC 87122202) is reacted with a 2-fold molar excess of D-biotinyl-ξ-amidocaproic acid-N-hydroxy-succinimide ester according to JACS 100 (1978), 3585-3590. Monobiotinylated IgG's are obtained as the main product and higher biotinylated side products which are separated off. 2) The separation is carried out by DIP chromatography (Delta-Isoelectric-Point) which is described e.g. in EP-A 0131 343 The separation of the mixture is carried out on a Mono-S cation-exchange column (Pharmacia). The mixture was applied in 1 mmol/l potassium pyrophosphate buffer pH 6.9 (buffer A). The bound components were eluted by the application of a linear gradient with 20 mmol/l potassium pyrophosphate buffer/200 mmol/l NaCl, pH 6.9 (buffer B). The pure monobiotinylated IgG fraction is obtained in a high yield as well as pools with a higher degree of biotinylation. Preparation of Fab-biotin (1:1) 1) Monoclonal antibodies against TSH (ECACC 87122202) are cleaved to Fab according to A. Johnstone, R. Thorpe; Immunochemistry in Practice, Blackwell Scientific Publications (1982), 52-53. 50 mg Fab are reacted as described in Example 4. 2) The cleavage is carried out here under modified conditions: Buffer A: 50 mmol/l MES pH 5.6 Buffer B: 50 mmol/l MES/200 mmol/l NaCl pH 5.6 Under these conditions pure Fab-biotin can be obtained in high yields. Abbreviations: Biotin X-OSu: N-biotinoyl-ξ-aminocaproic acid-hydroxysuccinimide ester MES: 2-(N-morpholino)-ethane sulfonic acid KPP: potassium pyrophosphate BSA: bovine serum albumin POD: peroxidase TSH test The test is carried out with a mixture of the two monoclonal antibodies against TSH (ECACC 87122201 and ECACC 87122202) which were prepared as Fab-biotin according to Example 5. Step 1: 3 μg Fab-biotin mixture in 1 ml 50 mmol/l phosphate buffer pH 7.5, 0.1% BSA are incubated for 2 hours with 200 μl sample (human serum supplemented with TSH) in a tube coated with streptavidin (Example 1, see AFP). Afterwards the unbound material is washed out. Step 2: 200 mU streptavidin-POD conjugate in 1 ml buffer (see above) is incubated for 1 hour and subsequently the unbound material is washed out. Step 3: 1 ml substrate solution (Example 1, see AFP) is incubated for 1 hour and the optical density is determined at 422 nm as a measure for the TSH content. Hepatitis B Virus Test This test used HBV-specific oligonucleotides of the known sequence of subtype adw (Ono Y. et al., Nucl. Acids Res. 11, 6: 1747-1757, hereby incorporated by reference). A. Digestion of HBV DNA-containing sera (lysis) 10 μl HBV containing serum is mixed with 10 μl of 0.2 NaOH and incubated for 1 hour at 37� C. and subsequently 30 μl neutralizing solution (100 mM KCl, 20 mM Tris/HCl, 3 mM MgCl2, 0.067N HCl) were added. An aliquot of this mixture is used in PCR amplifications. B. Amplification of HBV DNA 20 μl of sera lysed as above are used in PCR amplifications following the pattern presented below. ______________________________________Reagent or Sample Concentration of Final Concentration in PCRSample Storage Solution 20 μl/100 μl Batch______________________________________Primer 1 5 μM 200 nMPrimer 2 5 μM 200 nMdATP (Na salt) 4 mM 200 mMdCTP (Na salt) 4 mM 200 mMdGTP (Na salt) 4 mM 200 mMdTTP (Na salt) 4 mM 175 mMPCR Buffer 10 � 1 �Taq Polymerase 5 U/μl 2.5 U______________________________________ 10 � PCR Buffer: 100 mM Tris/HCl, 500 mM KC1, 15 mM MgCl2, 1 mg/ml gelatin, pH 9.0 Primer 1: Oligonucleotide of HBV sequence 2269-2289 Primer 2: Oligonucleotide of HBV sequence 2419-2439, reverse The PCR batches were amplified in a Perkin Elmer Thermal Cyler with 30 cycles of 30 sec at 92� C., 30 sec at 50� C. and 1 min at 70� C. C. Preparation of Monobiotinylated Detection Probes Using the HBVadw sequence 2332-2351 (detection probe 1) and the HBVadw sequence 2297-2316 (detection probe 2) oligonucleotides are prepared in a conventional DNA synthesizer incorporating, in one synthesis step, a commercially available biotinylated nucleotide. The sequences of the oligonucleotides correspond to the HBVadw sequence (Ono Y. et al., supra): purification was carried out according to standard methods. Alternatively, the oligonucleotides may be synthesized and subsequently biotinylated according to U.S. Pat. No. 4,711,955. In short, the method comprises mercuration, reaction with allylamine and biotinylation with biotinyl-N-hydroxysuccinimide ester (NHS-biotin). Subsequently, the oligonucleotides are subjected to gel electrophoresis to separate monobiotinylated product from compound conjugated to more than one biotin moiety. D. Denaturation and Detection of the DNA 100 μl of the PCR amplified mixture prepared according to step B are denatured and together with 400 μl hybridization buffer containing the two biotinylated HBV DNA detection probes according to step C, incubated in a tube coated with TRSA streptavidin (EP-A-O 344 578) for 3 hours at 37� C. The concentration of the biotinylated detection probes is 75 ng/ml each. The hybridization buffer contains 62.5 mM Na phosphate buffer, 0.94M NaCl, 94 mM Na citrate, 0.125% Na lauryl sarcosine, 0.6% bovine serum albumin, pH 6.5. After incubation, 3 washing steps using EnzymunR washing solution (Boehringer Mannheim) are carried out and subsequently 500 μl conjugate solution (50 mU/ml streptavidin POD conjugate (Sigma)) in 100 mM Tris/HCl, pH 7.5, 0.9% NaCl, 0.5% bovine serum albumin is added with a pipette. After incubation for 30 min at 37� C. another washing step using EnzymunR washing solution is carried out followed by incubation with substrate (ABTSR, Boehringer Mannheim) for another 30 min and subsequently the extinction is determined at 422 nm. Determination can be automated by means of an ES 300 R (by Boehringer Mannheim). The level of the signal detected is proportional to the HBV concentration of the sample. Patent Citations
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Steph Jones - Best New Songs, Albums, & Features
Listen to the best new songs and albums from Steph Jones as well as features written by the DJ Booth Staff. Last updated on February 13, 2011
Steph Jones
Southern Love
If many of your friends have November birthdays like mine, you'll have figured out by now that February is indeed a time for romance, love and…well, you get the point. While roses, chalk-flavored hearts and tacky...
Posted 4 years ago
Steph Jones
B E A utiful
Since Autotune-drenched club cut Galactic 2 hit our front page back in December of '09, up-and-coming singer Steph Jones has been a busy man – in addition to playing a major role on Breathing Underwater, an ABC...
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Steph Jones
Featuring: Audio Push
Galactic 2
When I was young, my father sat me down on his lap, looked me in the eye, and said "Richard, if you remember one thing I tell you, let it be this: the artist who rocks the dancefloor rules the world." OK, OK, I made that...
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Steph Jones Gives Lifetape Some "Gravity" [Download]
Los Angeles, Calif. -- Rising R&B star Steph Jones is back in the Booth to bring fans his latest mixtape – sorry, lifetape. Featuring Booth-premiered, reader-approved single "Up Up and Away" and much more, the currently...
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Steph Jones
Up Up and Away
Far from being intimidated by strong, assertive females, newly-independent R&B singer Steph Jones (formerly signed to Disturbing tha Peace) prefers his ladies empowered; super-empowered, if possible. Making its online...
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This past fall, Houston-native Steph Jones made his DTP debut on the collaborative track, Celebrity Chick, intended to kick off the still-unreleased DTP compilation album, Strength In Numbers. Now that the calendar has...
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Written on: 31/10/2012 by ExclusiveGraphics (1 review written)
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Watch Speedruns, Cure Cancer With the Awesome Games Done Quick Charity Marathon
Speedruns: The next best thing to becoming a doctor?
Games are finished with sonic speed, computer programs play games better than you, and cancer gets its butt kicked: Welcome to the 2015 edition of the Awesome Games Done Quick charity speedrun marathon to benefit the Prevent Cancer Foundation.
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Disney Princess Memory Game
Product information
This game features a set of 48 bright and colourful high-quality memory picture cards that feature lots of different images of kids' favourite Disney Princesses.
There are 24 different pairs to match up and collect in this classic game. The game play can be varied to suit different ages and abilities; for younger children use fewer cards, but you'll be surprised at just how good youngsters are at this game.
The memory cards are made from sturdy high-quality card, laminated on both sides with a colour print.>
- Brand
- Number of players
- 2 - 8
- Suitable From
- 3 years
- Toy Character
- Disney Princess
- Warning!
- Not suitable for children under 3 years
Brand Story
"
Walt and Roy Disney founded the 'Disney Brothers Cartoon Studio' on 16th October 1923 and it didn't take them long to establish themselves as leaders in the American animation industry. In later years, they branched out beyond this realm into live-action film production, television, and travel. The company is best known for its output from the infamous Walt Disney Studios, which is one of the largest and best-known studios in Hollywood today.

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Vic MP faces Ombudsman probe
PUBLISHED: 05 Jun 2012 16:06:00 | UPDATED: 07 Jun 2012 00:50:45PUBLISHED: 06 Jun 2012 PRINT EDITION: 06 Jun 2012A Victorian MP accused of misusing his electoral entitlements faces an investigation by the state's Ombudsman in a dramatic escalation of the scandal plaguing the Baillieu government.
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People | Ted Baillieu, George Brouwer |
Topics | Politics /Victorian Politics |


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AFTER a night of Christmas parties, it just wouldn't be complete without a drive through the Cape County Park Christmas lights display. Awesome.
ARE you kidding me? The city of Cape Girardeau is going to spend $1.6 million on expanding the trail at the river walk, but you can't fill the potholes on Lexington and William. My car is going to bottom out one of these days, and I am going to sue the city for it.
SINCE we are needing parking along Broadway, why don't we just go ahead and continue and start at Main Street and go west to Kingshighway and remove all buildings and make it a parking lot? That will give us ample parking.
YOU know what smokers really just don't get is the fact that their hair and their clothes -- no matter how new, no matter how beautiful they are -- stink. If they come in your house, they bring the stink right along with them. If you have a date and he smokes, he is going to stink and you are going to stink when you go home, too.
THE U.S. drone that was captured by Iran was not damaged because whoever was controlling it landed it there.
IF everyone would stop feeding the deer, the deer problem would go away. Half of the problem is that everyone has to feed the deer. The hunters want to feed the deer. All the people in town want to see the pretty deer. All the hunters want to keep the deer in their area so they can harvest the deer. The Department of Conservation is 10 years behind this problem. And as far as these expert bow hunters, all they want to do is kill deer. They can't be objective in any way.
I just completed reading the recent TBY insert. The article about a miracle, "We say yes" -- that was just precious, and it was so appropriate whenever we are thinking about the precious life of Jesus, who was born during this season. Thank you, Kim, for doing such a great job.
THE individual makes society, and society makes the individual. Do you see why we need ethical businessmen?
I think Rush Limbaugh has made a mistake calling people liberals. There are people in our culture who just exhibit bad behavior. Liberals are people who think and help develop society. They are trained at state universities, and a liberal Democrat is a good person. He believes in social justice, economic justice and so on in society.
I just read the front page story about the new initiatives for performance funding. They are supposed to be going forth in Missouri in which universities will be compensated depending on their performance. One of these things they are supposed to be trying to accomplish is improving student graduation rates. The problem with that is we are already graduating in this country lots of college graduates who can't write, who can't figure out how to read an article, who don't have the skills needed to succeed and compete with the rest of the world. This sort of thing is going to make it worse. I don't see this as progress. What we need to do is reward accomplishment, not just pass them through like the high schools have done for years.
HELP save a post office. Mail cards and letters.
CAPE Area Landlord Association, thank you for checking things and helping with the situation. Hopefully we can avoid a rental inspection program that costs us all more each year. Some of the houses that are being rented now are ones we lived in just a year ago. Nothing is wrong with them.
THIS is in reference to the Cape Girardeau School District possibly going to uniforms. Scott City did it. I was totally against it. But, after a few weeks or less, everybody was fine, everything was going along the way it needed to go, and you didn't see people dressed like tramps and thugs.
IF the Memphis, Tenn., inner city public schools can establish a dress code and have it work to eliminate bullying and some of the other problems in the school, any school in the United States could achieve this same situation. Money is not necessarily an object. If Memphis can do it, anybody can do it.
I'VE been reading and hearing that there is a public meeting on Dec. 29 at the Osage Centre. It's like a meeting to shut down the postal facilities. I urge everybody to be there: business people, people who do mailings, trucking companies and all other consumers. Everybody who mails letters and depends on the mail should attend because things are about to get real bad for the mail service. And that affects everyone in some way. It will also have a big impact on the local economy.


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Thanksgiving is almost upon us. And as we all fly and drive to towns far and away, prepare our epically long grocery lists or meticulously clean, disinfect and scrub our homes in preparation for the in-laws, we can get a wee bit stressed out. Will the turkey come out perfect? Will little Johnny and Jane sit still during dinner? Will Uncle Bill refrain from indulging in too much wine? So much to think about, so many anxious moments and so much worry. You know what we all need? A little comic relief, a little laugh, a giggle or perhaps a smirk, all in the name of Thanksgiving.
These twenty Thanksgiving photos represent a variety of funny, amusing and sometimes just sad turkey day fails. Just be thankful it didn't happen to you.
Check them out them right here:
Top Photo: TangoOrangers.com


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How Apple laptops interfaced with phones in 1999
1999 doesn't seem all that long ago, but it was -- it really was. Here we see a PowerBook 550c (1995/1996) getting frisky with an NTT Personal PALDiO (1999) Japanese flip phone in its PCMCIA slot. Not exactly as slick as an iPhone wirelessly chatting with a MacBook, but it was probably pretty damn cool at the time.
[Photo credit: raneko]
Software Updates
more updates- Apple Remote Desktop updated with Yosemite support
- OS X Yosemite 10.10.2, iOS 8.1.3 updates now available
- Sports Illustrated 120 SPORTS channel comes to Apple TV
- Logic Pro X update brings AirDrop support, new effects, tools, and more
- Parallels Access 2.5 released, adds file manager, computer-to-computer remote access
- The Google Translate iOS app is about to get a lot smarter


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Connecticut Baptist Homes has been compared
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Connecticut Baptist Homes in Meriden Connecticut has a 97% occupancy rate with 58 residents using its 60 beds. They are not part of a multiple nursing home ownership and are a Non Profit nursing home.
Connecticut Baptist Homes has an average registered nurse per resident per day of 50 minutes compared to the Connecticut state nursing home average of 50 minutes. They had 8 deficiencies in its past 2 state inspections compared to the CT average of 14 and complaints compared to the CT average of 6. Connecticut Baptist Homes had 3 fire safety deficiencies in the past 2 state inspections which is better than the Connecticut state nursing home average of 15.
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Hitwest.com is the 757269:th largest website within the world. The website is created in 28/09/2000, owned by unavailable, currently located in France and is running on IP 220.127.116.11 registered by GANDI SAS network. This site not uses Javascript for user interaction. This site uses CSS to manage the site layout. This site is running on the unavailable webserver. The server side programming lanquage of the site is unavailable. Hitwest.com Google Pagerank is 0 and it's domain is Commercial. Hitwest.com estimated worth is $5,276.77, with 1321 estimated visites per day and ad revenue of $ 3.96. -
Estimated Data
We are absolutely certain that every one is able to earn money from his website, Therefor we will display a short estimated numbers that might be achievable through dedication and seriousness work on your website.
Our estimations point that your Website Value is $5,276.77, Your Daily Visitors could be in the area of 1321 per day and your potential Daily Revenues could be around $3.96. -
Meta Data
Title: 1ere hit radio dans l Ouest, musique, jeux, webradio, videos, artistes | Hit West
Description: Ecouter la radio musicale Hit West, musique en ligne, clips videos, webradios 80, infos a Angers, Nantes, Rennes, Brest, Lorient, Bretagne
Keywords: Ere Hit Radio Ouest Musique Jeux Webradio Artistes West
Seo Perfomance
Alexa Rank: #757269
Links In: 176
Google Page Rank: 0
General Data
Created: 28/09/2000
Expires: 28/09/2019
Owner: unavailable
Hosted in: France
Host IP: 220.127.116.11
ICANN Registrar: GANDI SAS
Domain Suffix: com
Domain Archive: hitwest.com in the past
Technical Data
Server DNS A: 18.104.22.168
Server DNS NS: a.dns.gandi.net c.dns.gandi.net b.dns.gandi.net
Server Name: ouest-proxyvip2.sdv.fr
Server Type: unavailable
Server Side Language: unavailable
Javascript Usage: no
CSS Usage: yes
RSS Usage: no
Google AdSense Usage: no
Additional technologies usage: Google Analytics,jQuery,AddThis,GoogleFontApi
Ip Ranges Adress information
From - to Netname Description Country Admin Tech Status 188.8.131.52 - 220.127.116.11 SDVNET3 SdV Plurimedia 15 rue de la nuee bleue 67000 Strasbourg France --------------------------- For abuse: firstname.lastname@example.com --------------------------- FR DUMY-RIPE DUMY-RIPE ASSIGNED PA 220.127.116.11 - 18.104.22.168 FR-SDV-9900205 SdV-Plurimedia PROVIDER France FR DUMY-RIPE DUMY-RIPE ALLOCATED PA 126.96.36.199 - 22.214.171.124 EU-ZZ-212-213 RIPE NCC European Regional Registry EU DUMY-RIPE DUMY-RIPE ALLOCATED UNSPEC 0.0.0.0 - 255.255.255.255 IANA-BLK The whole IPv4 address space EU # Country field is actually a DUMY-RIPE DUMY-RIPE ALLOCATED UNSPEC -
Network Ip Calculator
- Address 184.108.40.206 11010100.01011111.01001000.00000100
- Netmask 255.255.255.0 = 24 11111111.11111111.11111111.00000000
- Wildcard 0.0.0.255 00000000.00000000.00000000.11111111
- Network 220.127.116.11/24 11010100.01011111.01001000.00000000
- HostMin 188.8.131.52 11010100.01011111.01001000.00000001
- HostMax 184.108.40.206 11010100.01011111.01001000.11111110
- Broadcast 22.214.171.124 11010100.01011111.01001000.11111111
- Hosts/Net 254 Class C
Traffic Data
Keywords Density
Keyword Count Density Hit 26 3.63 Vos 19 2.65 West 17 2.37 Jeux 16 2.23 Places 14 1.96 Ecouter 12 1.68 Replay 11 1.54 Infos 10 1.4 Gagnez 9 1.26 Gagner 7 0.98 Radio 7 0.98 Rhum 7 0.98 Route 7 0.98 Pour 6 0.84 édition 6 0.84 Concert 6 0.84 Fans 5 0.7 Nantes 5 0.7 Rennes 5 0.7 Une 4 0.56 Soir 4 0.56 -
More Websites on IP 22.214.171.124
Alternative Spellings
bitwest, ditwest, xitwest, hctwest, hftwest, hstwest, hitest, hitfest, hitiest, hitrest, hitwgst, hitwkst, hitwet, hitwect, hitwelt, hitweot, hitwevt, hitwext, hitwesc, hitwesf, fhitwest, ghitwest, shitwest, thitwest, hcitwest, hgitwest, hmitwest, hqitwest, htitwest, huitwest, hictwest, hiitwest, hiotwest, hirtwest, hitcwest, hitgwest, hitxwest, hitwiest, hitwjest, hitwpest, hitwuest, hitwvest, hitwekst, hitwevst, hitwesit, hitwesrt, hitwestd, hitwestq, hitwests, hitwestu,
Geo Data
Server Country Code: FR
Server Country Name: France
Server City Name: Strasbourg
Server Region Name: C1
Server Zip Code:
Server Latitude: 48.600399017334
Server Longitude: 7.7873997688293
Whois Data
Whois Server Version 2.0
Domain names in the .com and .net domains can now be registered
with many different competing registrars. Go to http://www.internic.net
for detailed information.
Domain Name: HITWEST.COM
Registrar: GANDI SAS
Whois Server: whois.gandi.net
Referral URL: http://www.gandi.net
Name Server: A.DNS.GANDI.NET
Name Server: B.DNS.GANDI.NET
Name Server: C.DNS.GANDI.NET
Status: clientTransferProhibited
Updated Date: 03-oct-2012
Creation Date: 28-sep-2000
Expiration Date: 28-sep-2019
>>> Last update of whois database: Fri, 31 Oct 2014 19:00:51 GMT <<<
NOTICE: The expiration date displayed in this record is the date the
registrar's sponsorship of the domain name registration in the registry is
currently set to expire. This date does not necessarily reflect the expiration
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registrar. Users may consult the sponsoring registrar's Whois database to
view the registrar's reported date of expiration for this registration.
TERMS OF USE: You are not authorized to access or query our Whois
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The Registry database contains ONLY .COM, .NET, .EDU domains and


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that would be money and timeSubmitted by B M (not verified) on Tue, 01/08/2013 - 9:43pm.
that would be money and time better spent, as the law stands now you can't even come on school grounds to pick up your children if you have a gun in your dash even though you're a ccw, it's time to update some out of touch laws, more law enforcement and bigger government hasn't worked, let people be responsible for their own safety for a change. »
Reply |


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28 Heartwarming Reasons Why Cosplayers Choose Their Characters
I asked attendees at L.A. Cosplay Con what drew them to the characters they chose. Here's what they said.
I asked attendees at L.A. Cosplay Con what drew them to the characters they chose. Here's what they said.
His appearance at New York Comic Con this past weekend was a huge relief for those of us who were a bit concerned at how damn skinny he was just over a year ago.
Seth Green shared a bonus Pacman sketch from the Season 6 Blu-ray (out on Oct. 8th) with BuzzFeed.
This poor guy needed help to fight the man! In Season Four of Robot Chicken, the episode titles told a story in production order, but the message got garbled when aired incorrectly.
Perfect parenting. Dan Milano of Robot Chicken took his toddler daughter with him to recreate the famous "Stay Puft" scene.
How do we feel about this? On the one hand, the super deformed character design is wonky but on the other hand, it's actually funny.
He's had it up to HERE with the jokes. And he's out for revenge.
You'll never look at the Justice League the same way again. Some stories can only be told through crass stop motion animation.
It featured Conan O'Brien, and predicted Britney Spears' eventual descent into crazyville. Those are two things we will applaud you for Mr. Green. Well done. View Media ›
Best. Panel. Ever. At the Robot Chicken panel a fan asked for co-creator Seth Green to oblige (for the third year in a row) with a sexy pose for a photo-op. And was then sexually assaulted assisted by Culkin.


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NPR just came out with an all-star mix featuring 100 hand-picked artists that should be on your radar during this year's SXSW.
This list includes Emmy The Great (pictured gloriously above), Cheerleader, Torres and Mitski!
Get the playlist HERE and make sure to swing by their performances during your trip in Austin!
Check out PHOX's stunning World Cafe session over at NPR!
Their self-titled debut is out now on Partisan Records.
Listen to Dirty Dishes on this week's episode of KEXP's Music That Matters! Every week KEXP chooses ten of their favorite tracks from new and independent artists around the globe.
Listen to "Guilty", and the rest of the mixtape, here!
Dirty Dishes is on tour NOW! Click here for a full list of dates.
Bob Boilen, host of NPR's All Songs Considered calls Monica Martin of PHOX, the band's "not-so-secret weapon", asserting that Martin "…sings with a smoky lilt in front of spare, tasteful instrumentation."
Need proof?
Go ahead and stream PHOX's 3-song performance on this week's episode of Boilen's Tiny Desk Concerts.
Set List - "Kingfisher," "1936," and "Slow Motion"
English synth-pop royalty aka Martin Gore of Depeche Mode announces that he'll be coming out with MG in April via Mute Records - an album idea he's been toying with ever since his DM-days. MG consists of 16 instrumental-only tracks, and the wait is (almost) over, so let's just cut straight to the source for details:
"I wanted to keep the music very electronic, very filmic and give it an almost sci-fi-like quality. Music is a necessity for me. I go into the studio at least five days a week, every week, so once I had the idea and the template, the process was quick and fun." - Martin Gore
Read more at Rolling Stone and listen to the album's lead single "Europa Hymn" streaming there now.
Check out Black Lips' former guitarist Ian St. Pé chat with Culturated.com about the newest album from alt-rock supergroup Diamond Rugs, tour plans, and his favorite color. Read the full interview here!
Cosmetics is spinning at radio NOW!
Glide Magazine has been added to the list of José González supporters for his new album Vestiges & Claws out now on Mute Records.
Below are some highlights from the review with the best tracks highlighted to check out first! Vestiges & Claws is going for spins at RADIO now!
- "What Will" … has the impact of a loud, crashing rock chorus.
- "Afterglow" with its' solemn hand-clapped percussion, has an aura of quiet abandonment.
- On "Stories We Build, Stories We Tell" finds Gonzalez contemplating the at-times selfish side of empathy. "Oh, they'll get to you," he sings. "Wondering what's on your mind; wondering what's wrong with you."
Check out the title track from Houndstooth's upcoming album, No News From Home via Pitchfork!
Houndstooth hail from Portland, Ore., but like most working indie bands, they spend much of their time on the road, far from the comforts of home. That sense of disconnection fuels the title track to their forthcoming second album, as singer/songwriter Katie Bernstein worries over an unresponsive pen pal whose silence leaves her feeling utterly untethered in the world. "I ride a spinning wheel everyday," she sings in a low, sad voice, then adds, "What can I say?"
Maybe it's not even news she wants, but something—anything—to alleviate the repetition of another long drive to another club show. If Bernstein strikes a melancholy tone, the rest of her band sound like they love the road, putting a little snap in the song. As John Gnorski unspools a rollicking guitar solo like he's flooring the gas, the rhythm section count the dotted yellow lines on the highway and put as many miles as possible between Bernstein and her worries.
No News From Home is going for adds on March 10!
Tune in or set your DVR TONIGHT at 12/10c PST to watch Brandi Carlile perform on Conan!
The Firewatcher's Daughter is spinning at radio NOW!
Behold Future Brown: the most innovative, geographic, time-and-genre-bending producers around! Combining forces from Los Angeles, New York, and London (by way of Kuwait), the debut LP from Future Brown (out now on Warp) takes listeners to another place in time.
Check out this awesome review from the writers at The Guardian!
The vocals… are what fix us in the present moment: the party talk, posturing and sexual provocations pose an interesting counterpoint to the sci-fi soundscapes.
Future Brown is at radio NOW!
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I believe in love, it's all we got
Love has no boundaries, costs nothing to touch
War makes money, cancer sleeps
Curled up in my father and that means something to me
Churches and dictators, politics and papers
Everything crumbles sooner or later
But love
I believe in love
I believe in love, it's all we got
Love has no boundaries, no borders to cross
Love is simple, hate breeds
Those who think difference is the child of disease
Father and son make love and guns
Families together kill someone
Without love
I believe in love
Without love I wouldn't believe
In anything that lives and breathes
Without love I'd have no anger
I wouldn't believe in the right to stand here
Without love I wouldn't believe
I couldn't believe in you
And I wouldn't believe in me
Without love
I believe in love
I believe in love
I believe in love
Writer(s): Elton John, Bernie Taupin
Copyright: Wab Publishing Ltd., Rouge Booze Inc.
Lyrics powered by www.musiXmatch.com
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12-02-1999 Comments (0)
Botulinum Toxin Type BCovera-HSMyoblocPrialtSchwarz PharmaVerapamilVerelan PMZiconotide
Elan Corp, based in Dublin, Ireland, has posted revenues of $214.9million and net income (before one-time…
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User Statshot
- Community Stats
Level 47 Blank Slate
Garbage Whistle
Ranked as Police Lieutenant
Latest News
Recent Game Medals
Question Mark
5 Points
Find the hidden button
Medal Stats.
The Flamer
25 Points
Get Fire Death
Medal Stats.
Pecking Order
25 Points
Get Crow Death
Medal Stats.
Kiss That Cyuke
25 Points
Get Rabbit Death
Medal Stats.
25 Points
Kill Hansel
Medal Stats.
Bitch Slap
10 Points
Shoot mom in the face
Medal Stats.
Get Stoned
50 Points
Get all 10 stones
Medal Stats.
Pest Removal
25 Points
Trap the squirrel
Medal Stats.
Sucked Dry
10 Points
Get Spider Death
Medal Stats.
Honey Horror
10 Points
Get Bee Death
Medal Stats.
Total Medals Earned: 70 (From 16 different games.)
Favorite Movies
Sketchy Stories
Rated 3.8319 Stars
A Young Boys Quest for IceCream
Comedy - Original
Dot Dot Dot - Animated
Rated 4.4526 Stars
An animation of a voice track of a review of a game.
Comedy - Original
I Own Time
Rated 4.0118 Stars
I Own Time
Comedy - Original
Awesome Reach
Rated 4.4327 Stars
Halo Reach is out. LOCK THE DOORS. CLOSE THE BLINDS. HOT POCKETS READY.
Comedy - Parody
Favorite Games
Vuvu Hero
Rated 4.1417 Stars
Do you have what it takes to be the vuvuzela hero?
Rated 4.1071 Stars
An arcade brawler where you can eat stuff to gain abilities
Action - Fighting - Brawler
Hedgehog Launch
Rated 4.4344 Stars
Your only opportunity to send hedgehog into orbit!
Skill - Toss
Dad 'n Me
Rated 4.2701 Stars
Chainsaw the Children guy's son goes on a rampage!!
Action - Fighting - Brawler
Favorite Audio
Alice is Dead | Indie Song | |
"Tetris" Remix | Video Game Song | |
Endless Handbag Loop | Video Game Loop | |
czer323 - Techno Fun5 Fixed | Techno Song |
Latest Audio Submissions
Red Sunday [Added Synth Mix] | Dance Song |
Red Sunday | Dance Song |
Loopy (FCM) | Techno Song |
Detonator | Techno Song |


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My first experience with creating help files came a fair few years ago, whilst eagerly reading a dusty Microsoft programming manual. I remember thinking that creating even the simplest form of online help seemed a mammoth task. Inserting appropriate tags here, using curious line breaks there, saving in special file formats somewhere else. What a load of tosh. But surely my experience was simply a result of living in pre-popular computing times, when Microsoft didn't even have a website and WYSIWYG was merely a new fangled toupee? Hmm, true, but even today most programmers avoid the task of creating their own help files. Way too difficult. Or is it? Before writing this review, I personally dabbled with a number of leading help file creation tools. Some were too expensive, others focused purely on the enterprise scene, and yet others were just too darn difficult. In this review, I'm taking a geek peek at EasyHelp, one of the elite couple that caught my eye. It claims to make writing a help file easier than, err, it was. So let's see how it measures up... The problem with many of the help programs I found was that they spent too much time trying to recreate an internal Word-like package that allowed the user to visually create help files. And that meant they forgot about silly little things, like usability. Thankfully EON Solutions didn't. In fact, their program doesn't even ship with an EXE. Instead, EasyHelp is a Microsoft Word add-in. And it's cunningly simple. My installation ate around 7MB of hard disk space, from a 30-day evaluation download of around 2.5MB. During setup, EasyHelp detects your copy of Microsoft Word and configures itself appropriately. And whether you're using Word 6, 7, 97 or 2000—or are attempting to create help files for Windows 3.1, 95, 98 or NT—you'll be in good company—all are supported. Creating a new help file is actually quite a doddle. You simply fire up Word, click File, New, then—instead of selecting 'Blank Document' or 'Professional Fax' as you typically would—choose the 'EasyHelp' template. After initially saving the document, you can start your future help file. Quite simply, you just start typing. Following my early experiences with the world's worst help file creators (a possible new Channel 5 documentary?)—this seemed too good to be true. Still, being the eternal optimist, I started. I tapped out a title, then a chunk of text for the body. Next, I started what I wanted to become the second page, plus added a little more writing. Next up, I clicked Process, which whizzes and whirs to zap out a help compatible RTF file. Finally, I pressed Build—which compiles and launches the final help file. And boy, was I impressed. In front of me, I found one long page containing all the text I'd tapped out. Sure, it wasn't Microsoft quality, but you've got to start somewhere. Hey, I'd actually created my own Windows help file! So there was my entire text on one whopping great page. Next up, it's recommended you split that mass of writing into individual topics. So perhaps 'Introduction', then 'Common Questions', maybe 'Support'. It's your call. Thankfully this is also exceptionally easy. I just selected the titles of what I wanted to become the individual topics—and clicked the Topic button. After specifying a name, that was it. I processed it all again—and EasyHelp had divided my test file up into two separate help file topics. Wow. Still, I thought, this beast must fall down somewhere. Nothing could be this easy. So I went ahead and started getting difficult. I added multi-coloured text, numerous mixed fonts, a hyperlink to my www.karlmoore.com homepage, a numbered list, bullet points—even a picture. And just to be extra troublesome, I added a link on the first topic across to the second, by clicking the Link button. Seconds later, I tested the help file once more. And by jove, everything worked. Everything! Of the many help file products I've seen, this one has to be the simplest. Creation consists of just a few easy-to-follow steps, with extra configuration options—such as title colours and searchable keywords—for power users. Definitely one of the market leaders and certainly worth the #x/$c registration cost. Whether you're building help files for Windows 3.1, 95, 98 or NT—if you're looking for the easiest solution, remember one name: EasyHelp. You can download a 30-day evaluation copy of EasyHelp from www.eon-solutions.com
Review of EasyHelp
November 19, 2002


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BEARdocs is currently undergoing a scheduled upgrade. We expect the upgrade to be completed no later than Monday, March 2nd, 2015. During this time you will be able to access existing documents, but will not be able to log in or submit new documents.
dc.contributor.advisor | Yelderman, Joe C. | |
dc.contributor.author | Diehl, Michelle Lynn. | |
dc.date.copyright | 2012-05 | |
dc.identifier.uri | http://hdl.handle.net/2104/8420 | |
dc.description.abstract | Management of groundwater resources is a critical issue in Texas and groundwater conservation districts have been given this responsibility. While large databases containing well characteristics are available for use from state agencies, they have not been organized for spatial correlation and analysis. As a new entity (2007), the Southern Trinity Groundwater Conservation District (STGCD) is faced with developing and analyzing such data. An unusual challenge for the STGCD is managing groundwater production among wells completed either solely in the upper aquifer (Hensell unit), lower aquifer (Hosston unit), or dually completed in both units of the Trinity Aquifer. The goals of this project were to develop a spatially-based well data set that can be used for management decisions. Results include a report to the STGCD with a database, contour maps for different aquifer characteristics, and well hydraulics analysis. | en_US |
dc.publisher | en | |
dc.rights | Baylor University theses are protected by copyright. They may be viewed from this source for any purpose, but reproduction or distribution in any format is prohibited without written permission. Contact firstname.lastname@example.com for inquiries about permission. | en_US |
dc.subject | Groundwater | en_US |
dc.subject | Aquifers | en_US |
dc.subject | Trinity Aquifer, Central Texas | en_US |
dc.subject | Southern Trinity Groundwater Conversation District | en_US |
dc.title | Intra-aquifer characterization and potential management impacts : Trinity Aquifer, Central Texas. | en_US |
dc.type | Thesis | en_US |
dc.description.degree | M.S. | en_US |
dc.rights.accessrights | Worldwide access | en_US |
dc.contributor.department | Geology. | en_US |
dc.contributor.schools | Baylor University. Dept. of Geology. | en_US |


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Definition of disinterestedness
n. - The state or quality of being disinterested; impartiality.
The word "disinterestedness" uses 17 letters: D D E E E E I I N N R S S S S T T.
No direct anagrams for disinterestedness found in this word list.
Adding one letter to disinterestedness does not form any other word in this word list.
Words within disinterestedness not shown as it has more than seven letters.List all words starting with disinterestedness, words containing disinterestedness or words ending with disinterestedness
All words formed from disinterestedness by changing one letter
Other words with the same letter pairs: di is si in nt te er re es st te ed dn ne es ss
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Previous word in list: disinterestedly
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We're sorry, we need more information to run your search. Please enter a last name (adding more details will improve your results).
Locating the specific Adena Shemberg you are looking for is easy with PeopleFinders.com. We have a wide variety info for your people search such as age, recent addresses, and phone numbers. For example, Adena Shemberg was born in [YOB] and is 42 years old. The current address for Adena Shemberg is located in Great Neck, NY.
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Iggy Pop, Tommy Lee Help Sum 41 Frontman Get Sober
Deryck Whibley has been able to stay sober thanks to the help of a few classic rockers.
Deryck Whibley has been able to stay sober thanks to the help of a few classic rockers.
Safety reasons will prevent Tommy Lee's 'Cruecifly' drum coaster from being performed at every concert on Motley Crue's 'Final Tour.'
Nobody in rock history has done more to expand the concept of what a drum solo can look and sound like than Motley Crue's Tommy Lee. Here's video proof.
Tommy Lee talks about Motley Crue's upcoming song, which may or may not be the last track they ever release.
Billy Corgan explains his decision to hire Tommy Lee for the new Smashing Pumpkins album by comparing him to the Led Zeppelin drummer.
Frontman Billy Corgan says Tommy Lee appears on all nine songs from the Smashing Pumpkins' forthcoming 'Monuments To An Elegy.'
Tommy Lee might be saying goodbye to life on the road with Motley Crue, but he's saying hello to the married life.
A judge has ruled that Tommy Lee's 360-degree roller-coaster drums were based on his original concept. According to TMZ, the Motley Crue drummer designed the kit, which an engineer claimed was stolen from his 20-year-old idea.
MTV may not play music videos anymore, but Fall Out Boy haven't forgotten about the art of adding a compelling visual component to a song. In fact, they've filmed a video for every track on their latest album, 'Save Rock and Roll' -- and the newest clip features a cameo from Tommy Lee as Satan.
The countdown to what's being touted as Motley Crue's farewell tour has officially begun. But details about the exact reach, dates and reasons behind the decision remain subject to wild debate.


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We found 2 sites within 15 miles of Roscrea with 1 bookable
Ballinsloe, Co. Galway, Republic of Ireland Show on map |
Non-electric grass pitch | 6 x | From £11.82/€15/AU$21.71/$19.03 per night |
The Apple Camping and Caravan Park Cahir, Co. Tipperary, Republic of Ireland Show on map |
Want to see more? We're only showing the sites that are bookable on Pitchup.com.
Show campsites and holiday parks that aren't bookable through Pitchup.com...


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Get Fit Plus fitness studio in Watertown CT features many different programs to transform your body, your mind, and your soul! Fitness and health are a way of life and that is what you will experience at Get Fit Plus! We are a fitness family all on the path of living healthy lifestyles. When you step into our studio you will receive the attention that you need, support and motivation, and a sense of accomplishment every time you leave!
At Get Fit Plus we are NOT a gym! We are a private fitness studio that is focused on getting you results! Our programs can be made to fit any goal and any ability level! No cookie cutter programs found here. We pride ourselves on really knowing each and everyone of our members and what they are trying to achieve so we can make sure you see success!
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Get Fit Plus works with the Watertown Youth Service Bureau to help support the developing our youth! This is a wonderful program that helps direct kids to the proper resources available in this community. To learn more about this Organization click here. To learn more about our Check in for charity program click here!
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January 10th Starts Running Training For Shamrock Shuffle
January 30th and 31st Next Rank Up Days!


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John Legend Launches Worldwide Concert Tour in November in Support of "Evolver"
New York, NY -- Multi-platinum Grammy-Award-winning recording artist and acclaimed performer John Legend will embark on an extensive worldwide concert tour kicking off November 19 in Minneapolis. The first leg of John's North American tour wraps up December 13 with a second leg running from January 12 through February 9. (Please see John Legend tour itinerary below.)
The upcoming John Legend concerts mark the artist's first major tour since his massively successful world tour in 2007 (captured on Live From Philadelphia CD and DVD).
John's special guest on the tour's initial dates is his Columbia Records label mate Raphael Saadiq, who will be performing songs from his just-released new album, The Way I See It.
One of the most eagerly-awaited releases of 2008, EVOLVER is John Legend's first full-length studio album in two years and will be released October 28. Guests on EVOLVER include OutKast's Andre 3000, emerging hip-hop/soul ingénue Estelle, R&B/pop superstar Brandy and John's longtime collaborator Kanye West.
* * * * *
John Legend - 2008 Tour Dates
Wed 11/19 Minneapolis, MN Northrop Auditorium
Thu 11/20 Chicago, IL Chicago Theatre
Mon 11/24 E. Lansing, MI Wharton Center For Performing Arts
Wed 11/26 Detroit, MI Masonic Temple Theatre
Fri 11/28 Toronto, Ont. Roy Thompson Hall
Sat 11/29 Montclair, NJ Wellmont Theatre
Mon 12/1 Montclair, NJ Wellmont Theatre
Tue 12/2 Boston, MA Orpheum Theatre
Fri 12/5 Philadelphia, PA Tower Theater
Mon 12/8 Baltimore, MD Lyric Theatre
Tue 12/9 Washington, DC Constitution Hall
Wed 12/10 Greenville, SC Peace Center
Fri 12/12 Charlotte, NC Ovens Auditorium
Sat 12/13 Durham, NC Durham Performing Arts Center
John Legend Raphael Saadiq Concerts
Written by richard on 10/7/08
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(Prior to Fall 2010, this course was known as HIST 13.The information below might still reflect the old course numbers. Bracketed numbers, if any, are the old course numbers. Learn more...)
3 hours; 3 credits American history to 1877. Political and economic developments from the colonial origins of American institutions through the Revolutionary era and the periods dominated by Federalism, Jeffersonianism, and Jacksonianism. Consideration of the issues of the Civil War and the Reconstruction era. Satisfies Pathways Flexible Core US Experience in Its Diversity requirement.


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It's their time to shine on the big stage and prove that they can play at that level."
That's Furman coach Bobby Lamb talking about his team's game this week at Missouri, of the Big 12 Conference.
Furman and two other Palmetto State FCS squads will get their moment on the big stage this weekend, as Wofford travels to Wisconsin, of the Big Ten, and Charleston Southern goes back to the state of Florida to play South Florida, of the Big East.
Which of the three teams has the best shot at pulling off an upset? Glad you asked. In order of likelihood:
-- Wofford at Wisconsin -- The last time Wisconsin played a Southern Conference team, the 7th-ranked Badgers were booed off the field at halftime by their own fans, tied with The Citadel at 21-21.
Wisconsin went on to win that 2007 game, 45-31, and Badgers coach Bret Bielema is sure to remind his players that the Terriers are from the same league as the Bulldogs.
The Badgers are 2-0 this season and needed double-overtime for a 34-31 win over Fresno State last Saturday. Wisconsin had more than 40 players suffering from flu-like symptoms last week, with at least six getting IV fluids before the game.
Wisconsin also needed OT to get past FCS squad Cal Poly, 36-35, last season, and a flu-weakened squad could be ripe for a Wofford upset.
Unfortunately, the Terriers have their own problems as their leading rusher, fullback Eric Breitenstein, is out four to six weeks with a knee injury suffered last week against Charleston Southern.
Advice to Wofford -- pack some face masks to wear under your face masks.
--Furman at Missouri -- The Paladins are playing the best football of the state's FCS schools at the moment, at 2-0 with wins over Presbyterian and Chattanooga. Furman quarterback Jordan Sorrells has been sharp, and the Paladins have a nice run-pass balance even without tailback Mike Brown (ankle).
Missouri is 2-0, but this does not seem to be quite the Mizzou team of recent vintage that featured quarterback Chase Daniel and receiver Jeremy Maclin.
The Tigers opened with a 37-9 win over Illinois, but struggled last week in a 27-20 victory over Bowling Green, leaving some Missouri fans uneasy.
That should give Mizzou coach Gary Pinkel plenty of ammo to keep the Tigers focused this week.
"Our football team didn't get better from week one to two, and that's disappointing to me," Pinkel said. "That's my job … I'm going to do my best to get that fixed."
Missouri has averaged a 43-7 score against FCS teams under Pinkel, but Furman has a history of playing the big boys much tougher than that.
"The closer we can get to the fourth quarter with some sort of chance, the pressure goes on Missouri," Lamb said.
--Charleston Southern at South Florida -- CSU coach Jay Mills was encouraged after a 62-3 loss at No. 1 Florida to open the season, but disappointed after last week's 42-14 loss to Wofford.
He might also have issues with whomever scheduled three straight road games at Florida, Wofford and Big East contender South Florida to start the season.
"It's a challenging schedule, probably the most challenging in FCS to start the season," Mills said. "But we felt like it would be beneficial to our program, and we feel that way in hindsight as well."
Comparative scores don't hold out much hope for the 0-2 Bucs against 2-0 South Florida, which beat Wofford 40-7.
The Bulls feature a "Toro Loco" offense that has QB Matt Grothe lined up at receiver, and receiver B.J. Daniels behind center. Grothe's block sprung Daniels for a 7-yard TD run in a 35-13 win over Western Kentucky last week.
And if the Bulls lose to CSU, the Toros really will be Loco.
Reach Jeff Hartsell at email@example.com.
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The A-6 Intruder was an American, twin jet engine aircraft. Operated with the U.S. Navy and U.S. Marines between 1963 and 1997 when they where retired from service. Exposed at Intrepid Sea, Air an... (more)
This is probably the most fantastic terminal I've ever seen. Is an open space terminal only closed with security controls. From the road you can see the hole terminal, and once you pass the securi... (more)
About to land at AEP.
Leaving the tarmac on their way back to the USA.
Heading to the runway.
Standing at Fiumicino and getting ready for the next flight.
Taking a close look, there's a bump on to of the cockpit. Probably not big deal. Here we can see Hotel Juliet on the starting engines' procedure at CDG.
On pushback from the terminal at BIO.
Landing on runway 27 at Heraklion.
Only 201 planes were built of the Tiger model between 1957-58. Between 1958 to 1960 the squadron VF-33 flew Tigers off the Intrepid. Here we can see 141884 / 5 painted on Blue Angels colours on st... (more)
On taxi to the runway.
While the plane gets to the parking, the captain saw me a did the victory sing with his finguers. Thank's captain.
Getting to the parking.
Heading to the parking area at Glasgow.
On taxi out for departure.


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Oracle Java SE CVE-2014-0448 Remote Security Vulnerability
Oracle Java SE is prone to a remote security vulnerability in Java SE. The vulnerability can be exploited over multiple protocols. This issue affects the 'Deployment' sub-component. This vulnerability affects the following supported versions: Java SE 7u51, Java SE 8 |
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Categories 7-Day Forecast First Alert Doppler Weather Cameras Humidity 96% Winds 4.6mph View Full Forecast › 21° Colorado Springs ›
Categories 7-Day Forecast First Alert Doppler Weather Cameras Humidity 92% Winds 11.5mph View Full Forecast › 25° Pueblo ›
Union says Denver officers ordered not to wear riot gear DENVER (AP) - Denver's police union says officers' safety is being compromised by orders not... Read Story ›
Thursday Weather: Sunny & Warm (Rinse & Repeat) Colorado Springs hit 60 degrees on Wednesday afternoon. Pueblo warmed to 62 degrees. Those Wednesday... Read Story ›
UNLV hires high school football coach LAS VEGAS (AP) - Bishop Gorman High School coach Tony Sanchez is set to take... Read Story ›
Slum study sought for Lower Westside PUEBLO - Pueblo City Council is expected vote tonight on proposed resolution asking the Pueblo... Read Story ›
Pueblo County records first death from flu A person has died from influenza in Pueblo County, according to Public Health officials. 60... Read Story ›
Union says Denver officers ordered not to wear riot gear DENVER (AP) - Denver's police union says officers' safety is being compromised by orders not... Read Story ›
2 months 3 weeks ago Flood Mitigation Projects to Start Monday in Cheyenne Cañon Dozens of flood mitigation projects are finally wrapping up in Colorado Springs, since heavy rains and floods did major damage to parks, trails and roads in September 2013. The work starts Monday in Cheyenne Cañon and will last anywhere from eight to 12 weeks, depending on the weather. Some trails... Read Story › Share: Facebook Twitter Google Plus
5 months 2 weeks ago First part of massive Williams Canyon drainage project nears completion The first part of a massive project in Williams Canyon is almost complete. News 5 got an inside look at the progress and what's next for the new drainage structure that's meant to prevent devastating flooding like we saw about a year ago in Manitou Springs. "The guys working... Read Story › Share: Facebook Twitter Google Plus
1 year 5 days ago Waldo Flood Mitigation Intends to Slow Down Flood Waters and Catch Debris Work is being done in the Waldo Canyon burn area to prevent devastating flooding like El Paso County saw last year. Most people are aware of the culvert being installed along Highway 24 to funnel water under the highway, but there's much more going on in the area. New... Read Story › Share: Facebook Twitter Google Plus
1 year 10 months ago Mayor wants to dip into saving to cover flood needs COLORADO SPRINGS - Mayor Steve Bach is asking the Colorado Springs City Council to take $11,875,000 million dollars out of savings. A majority of the money, $10 million worth, is directly tied to flood and fire mitigation efforts. The rest would be spent on a study to see whether... Read Story › Share: Facebook Twitter Google Plus
1 year 10 months ago Mayor's office seeks $10M for flood mitigation COLORADO SPRINGS - The Mayor's office plans to ask City Council for an additional $10 million this year to pay for flood mitigation work in and down stream from the Waldo Canyon burn area. The money would come from the city's reserves and a majority of it, $8.8 million, would... Read Story › Share: Facebook Twitter Google Plus
Play Black Forest Selects New Fire Chief Play Air Academy Hockey, Athlete of the Week Play Thousands of kids in need this Christmas Play Improving Stormwater drainage in Colorado Springs Play K9 coming to Canon City Police Dept. Play Report released about safety lapse in Pueblo abuse case Play Pit Bull Trial Play Your Healthy Family Play Pit bull released and reunited with his family Play Caught On Cam: Fiery Rescue Play Santa Delivers A Miracle Play New hospital gown gives full coverage Play City Council and Mayor Bach Battle over Budget Play Urban Renewal hopeful for city loan Play PD: Vandals do over $40K in damage to Pueblo school Play StopFraudColorado.gov targets scammers 1 2 3 4


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A dozen daily deals, day four
The Daily Deals just keep on coming from our friends at DealNews.com. Even though today is Sunday, you still have time to hit the Internets and pick up some more great deals for all your holiday shopping needs. After all, there are only 25 shopping days until Christmas, so you better get busy.
Here are the deals for today:
MacMall: [Cyber Monday] MacMall 3-Day Apple Cyber Monday Sale: Up to 25% off Apple systems, more after rebate
Staples: [Cyber Monday] Staples Cyber Monday Savings Event available online
Adorama: [Cell Phones] Unlocked Sony Ericsson W350A Walkman Phone for $70 + free shipping
Best Buy: [USB Hard Drives] Seagate Expansion 500GB USB 2.0 External Hard Drive for $50 + $6 s&h
Staples: [USB Hard Drives] Seagate Expansion 2TB USB 2.0 External Hard Drive for $140 + free shipping
TigerDirect: [Projectors] 3M MPro110 LED Pico Projector for $155 + free shipping
Walmart: [Office Furniture] Orion L-Shaped Desk with Hutch for $89 + $29 s&h
Staples: [22-Inch LCD] Acer 22" 1080p Widescreen LCD Display w/ HDMI for $110 + free shipping
Dell Home: [20" - 22" LCD TVs] Vizio 20" 720p Widescreen LCD HDTV for $165 + free shipping
ZipZoomFly: [USB Drive Enclosures] Thermaltake BlacX eSATA / USB 2.0 Docking Station for $20 after rebate + free shipping
Best Buy: [Plasma TVs] Panasonic Viera TC-P58S1 58" 600Hz 1080p Plasma HDTV for $1,480 + free shipping
6ave: [40" - 42" LCD TVs] LG 42" 120Hz 1080p LCD HDTV w/ NetFlix, YouTube for $788 + free shipping
Here are the deals for today:
Software Updates
more updates- Apple Remote Desktop updated with Yosemite support
- OS X Yosemite 10.10.2, iOS 8.1.3 updates now available
- Sports Illustrated 120 SPORTS channel comes to Apple TV
- Logic Pro X update brings AirDrop support, new effects, tools, and more
- Parallels Access 2.5 released, adds file manager, computer-to-computer remote access
- The Google Translate iOS app is about to get a lot smarter