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US 7601846 B2 Abstract The present invention provides compounds that can protect mammalian cells from the damaging effects of chemotherapy, irradiation, or in other situations in which it is desirable to protect tissue from the consequences of clinical or environmental stress.
Claims(4) 1. A compound of formula (IA):
wherein R3 is hydrogen, halo, hydroxy, cyano, �N(Ra)(Rb), �S(Ra), �NO2, (C1-C6)alkyl, (C1-C6)alkoxy, (C2-C6)alkynyl, (C2-C6)alkenyl, (C2-C7)alkanoyl, (C2-C7)alkanoyloxy, or (C3-C7)cycloalkyl, wherein Ra and Rb are each independently hydrogen, (C1-C3)alkyl, (C2-C4)alkanoyl, phenyl, benzyl, or phenethyl; or Ra and Rb together with the nitrogen to which they are attached are a 5-6 membered heterocyclic ring;
wherein R4 is �N(Ra)(Rb) and wherein the �N(Ra)(Rb) of R4 is a 5-6 membered heterocyclic ring, optionally comprising 1, 2, or 3 N(Ra), non-peroxide O or S atoms;
Y is �S(O)0-2; n is 1, 2, 3, 4, or 5; or
a pharmaceutically acceptable salt thereof.
2. The compound of
3. The compound of
4. A method for protection of mammalian cells from the damaging effects of chemotherapy, or irradiation, comprising administering to a mammal in need of said protection an effective amount of a compound of
Description This application is a continuation-in-part of U.S. patent application Ser. No. 11/077,986, filed Mar. 11, 2005 now abandoned, which is a continuation application of U.S. patent application Ser. No. 10/364,663, filed Feb. 11, 2003 now abandoned, which application is a continuation under 35 U.S.C. 111(a) of PCT/US01/25175, filed Aug. 10, 2001, which claims priority to U.S. patent application Ser. No. 09/637,531, filed Aug. 11, 2000, and U.S. Provisional Patent Application Ser. No. 60/301,340, filed Jun. 26, 2001, all of which are incorporated by reference herein. This invention was made with the assistance of the National Institutes of Health under Grant Nos. GM23200 and CA81534. The U.S. Government has certain rights in this invention. The prevention of undesired cell death has become an important goal for pharmacologic intervention in a variety of clinical settings. Profound disability can result from apoptotic cell death and tissue injury due to ischemia, chemotherapeutic agents, ionizing radiation, or hyperthermia. Recently, a small molecule, 2-(2-imino-4,5,6,7-tetrahydro-benzothiazol-3-yl)-1-p-tolyl-ethanone hydrobromide (PFT-α; 1), However, the activity of PFT-α is not limited to inhibiting the transactivation of p53. This small molecule has been found to also suppress the heat shock and glucocorticoid signaling pathways Komarova, E. A., et al., J Biol Chem. 2003, 278, 15465-15468. Glucocorticoid-induced cell death is independent of p53 Clarke, A. R., et al., Nature 1993, 362, 849-852. This pathway, however, also induces the transcription and activity of the proapoptotic BH3 (Bcl-2 Homology)-only protein p53-upregulated modulator of apoptosis (PUMA) Han, J.-W.; Flemington, C. et al., Proc Natl Acad Sci USA. 2001, 98, 11318-11323. When this death-inducing signal reaches the mitochondria, a series of cell-death related events ensues. The inner mitochondrial membrane loses its potential and cytochrome c is released into the cytoplasm, where it can associate with ATP, apoptosis-activating factor-1, and procaspase-9. This apoptosome complex cleaves procaspase-9 to caspase-9, which in turn cleaves procaspase-3, initiating the cascade of protease activation in the execution phase of apoptosis Li, P., et al., Cell 1997, 91, 479-489. and Zou, H., et al., Cell 1997, 90, 405-413. PFT-α has been reported to diminish p53-dependent and independent mitochondria mediated cell death in vitro and in vivo, Komarov, P. G.; et al., Science 1999, 285, 1733-1737 and Komarova, E. A., et al., J Biol Chem. 2003, 278, 15465-15468. The multiplicity of molecular pathways that are reportedly influenced by PFT-α suggests that the cytoprotective effect could be further optimized. Furthermore, PFT-α is not stable under physiologic conditions and spontaneously undergoes ring closure to form the imidazo[2,1-b]benzothiazole (IBT) 2, Singh, A.; Mohan, J., et al., Indian J. Chem., Sect. B 1976, 14B, 997-998. Thus, we report the synthesis and structure activity relationship of novel derivatives and analogs of PFT-α, and corresponding closed ring counterparts, with enhanced potency and stability. To determine activity, we used a p53-independent assay of cell death wherein mouse thymocytes were treated with dexamethasone in the presence or absence of test compounds. In addition, we confirmed the activity of the most potent compounds in a p53-dependent apoptosis assay. In addition, there is a need for novel, potent, and selective agents to prevent detrimental effects upon cells due to DNA damage, such as caused by chemotherapy, radiation, ischemic event, including ischemia-reperfusion injury and organ transplantation, and the like. There is also a need for pharmacological tools for the further study of the physiological processes associated with intracellular DNA damage. A continuing need exists for compounds that can protect mammalian cells from the damaging effects of chemotherapy and irradiation, or in other situations in which it is desirable to protect tissue from the consequences of clinical or environmental stress. The present invention provides compounds that can protect mammalian cells from the damaging effects of chemotherapy and irradiation, or in other situations in which it is desirable to protect tissue from the consequences of clinical or environmental stress. Accordingly, there is provided compounds of formula (I) and a method of protecting the cells comprising administering to a mammal in need of said suppression an effective amount of a compound of formula (I): Ar is aryl, heteroaryl, or a 5-6 membered heterocyclic ring, preferably comprising 1-3 N(Ra), non-peroxide O or S atoms, such as a pyrrolidino, piperidino or morpholino ring, optionally substituted with 1-5, preferably 1-2, halo, CF3, hydroxy, CN, N(Ra)(Rb), (C1-C6)alkyl, (C1-C6)alkoxy, (C2-C7)alkanoyl, (C2-C7)alkanoyloxy, (C3-C7)cycloalkyl, (C2-C6)alkanoyl, (C2-C6)alkenyl, or phenyl; Y is oxy (�O�), S(O)0-2, Se, C(R1)(R3), N(Ra), or �P�; or a pharmaceutically acceptable salt thereof. Preferably, Ar is not substituted with halo or alkoxy. Preferably, Ar is heteroaryl or a heterocyclic ring. Preferably, R1 and R2 are not benzo or (C3-C5)alkylidenyl when Ar is aryl, e.g., is phenyl or napthyl. Novel compounds of formula (I) are also within the scope of the present invention, e.g., preferably Y is �O�, �Se�, C(R1)(R3) or P. Preferably, Ar is heteroaryl. Preferably, Ar is substituted with CN, (C2-C7)alkanoyl), (C2-C7)alkanoyloxy, (C3-C7)cycloalkyl, (C2-C6)alkenyl or combinations thereof. Preferably, R1, R2 and R3 are independently, OH, CN (N(Ra)(Rb), S(Ra), NO2, (C2-C7)alkanoyl, or (C2-C7)alkanoyloxyl. In a specific embodiment the compounds of the invention have formula (IA): Additionally, the invention provides a therapeutic method for preventing or reducing the damaging effects of chemotherapy and irradiation, or in other situations in which it is desirable to protect tissue from the consequences of clinical or environmental stress comprising administering to a mammal in need of such therapy, an effective amount of one or more compounds of formula (I), or a pharmaceutically acceptable salt thereof. Such pathological conditions or symptoms include blocking, moderating or reversing the deleterious effects of chemotherapeutic agents, particularly those which damage DNA; radiation, particularly radiation therapy (gamma-, beta- or UV-radiation), ischemic event, including stroke, infarct, ischemia-reperfusion injury and ischemia due to organ, tissue or cell transplantation; environmental pollution or contamination and the like. The invention provides pharmaceutical compositions comprising novel compounds of formula (I), or a pharmaceutically acceptable salt thereof, in combination with a pharmaceutically acceptable diluent or carrier. The invention provides a compound of formula (I) for use in medical therapy as well as the use of a compound of formula (I) for the manufacture of a medicament for the treatment of a pathological condition or symptom in a mammal, such as a human, induced cellular damage, i.e., with unwanted apoptosis. The invention also includes a method for binding a compound of formula (I) to cells and biomolecules comprising p53 or p53 dependent receptors, in vivo or in vitro, comprising contacting said cells or biomolecules with an amount of a compound of formula (I) effective to bind to said receptors. Cells or biomolecules comprising ligand-bound p53 or p53 dependent receptor sites can be used to measure the selectivity of test compounds for specific receptor subtypes, or can be used as a tool to identify potential therapeutic agents for the treatment of diseases or conditions associated with p53 dependent activation, by contacting said agents with said ligand-receptor complexes, and measuring the extent of displacement of the ligand and/or binding of the agent, by methods known to the art. The following definitions are used, unless otherwise described: halo is fluoro, chloro, bromo, or iodo. Alkyl, alkoxy, alkenyl, alkynyl, etc. denote both straight and branched groups; but reference to an individual radical such as �propyl� embraces only the straight chain radical, a branched chain isomer such as �isopropyl� being specifically referred to. Aryl denotes a phenyl radical or an ortho-fused bicyclic carbocyclic radical having about nine to ten ring atoms in which at least one ring is aromatic. Heteroaryl encompasses a radical attached via a ring nitrogen or carbon of a monocyclic aromatic ring containing five or six ring atoms consisting of carbon and one to four heteroatoms each selected from the group consisting of non-peroxide oxygen, sulfur, and N(X) wherein X is absent or is H, O, (C1-C4)alkyl, phenyl or benzyl. Heteroaryl also includes a radical of an ortho-fused bicyclic heterocycle of about eight to ten ring atoms, particularly a benzo-derivative or one derived by fusing a propylene, trimethylene, or tetramethylene diradical thereto. Preferred heteroaryls include pyridin-4-yl and thiophen-2-yl. The term �heterocyclic ring� �heterocycle,� or �heterocycyl,� is defined as above for formula (I). It will be appreciated by those skilled in the art that compounds of the invention having a chiral center may exist in and be isolated in optically active and racemic forms. Some compounds may also exhibit polymorphism. It is to be understood that the present invention encompasses any racemic, optically active, polymorphic, or steroisomeric form, or mixtures thereof, of a compound of the invention, which possess the useful properties described herein, it being well known in the art how to prepare optically active forms (for example, by resolution of the racemic form by recrystallization techniques, by synthesis from optically active starting materials, by chiral synthesis, or by chromatographic separation using a chiral stationary phase) and how to determine cell protection activity using the standard tests described herein, or using other similar tests which are well known in the art. Specific and preferred values listed below for radicals, substituents, and ranges, are for illustration only; they do not exclude other defined values or other values within defined ranges for the radicals and substituents. Specifically, (C1-C6)alkyl can be methyl, ethyl, propyl, isopropyl, butyl, iso-butyl, sec-butyl, pentyl, 3-pentyl, or hexyl; (C3-C7)cycloalkyl can be cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl; the term cycloalkyl includes (cycloalkyl)alkyl of the designated number of carbon atoms; (C3-C5)cycloalkyl(C2-C4)alkyl can be cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl, 2-cyclopropylethyl, 2-cyclobutylethyl, 2-cyclopentylethyl, or 2-cyclohexylmethyl; (C1-C6)alkoxy can be methoxy, ethoxy, propoxy, isopropoxy, butoxy, iso-butoxy, sec-butoxy, pentoxy, 3-pentoxy, or hexyloxy; (C2-C6)alkenyl can be vinyl, allyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, or 5-hexenyl; (C2-C6)alkynyl can be ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl, or 5-hexynyl; (C2-C7)alkanoyl can be acetyl, propanoyl or butanoyl; halo(C1-C6)alkyl can be iodomethyl, bromomethyl, chloromethyl, fluoromethyl, trifluoromethyl, 2-chloroethyl, 2-fluoroethyl, 2,2,2-trifluoroethyl, or pentafluoroethyl; hydroxy(C1-C6)alkyl can be hydroxymethyl, 1-hydroxyethyl, 2-hydroxyethyl, 1-hydroxypropyl, 2-hydroxypropyl, 3-hydroxypropyl, 1-hydroxybutyl, 4-hydroxybutyl, 1-hydroxypentyl, 5-hydroxypentyl, 1-hydroxyhexyl, or 6-hydroxyhexyl; (C1-C6)alkoxycarbonyl can be methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, pentoxycarbonyl, or hexyloxycarbonyl; (C1-C6)alkylthio can be methylthio, ethylthio, propylthio, isopropylthio, butylthio, isobutylthio, pentylthio, or hexylthio; (C2-C6)alkanoyloxy can be acetoxy, propanoyloxy, butanoyloxy, isobutanoyloxy, pentanoyloxy, or hexanoyloxy; aryl can be phenyl, indenyl, or naphthyl; and heteroaryl can be furyl, imidazolyl, triazolyl, triazinyl, oxazoyl, isoxazoyl, thiazolyl, isothiazoyl, pyrazolyl, pyrrolyl, pyrazinyl, tetrazolyl, pyridyl, (or its N-oxide), thienyl, pyrimidinyl (or its N-oxide), indolyl, isoquinolyl (or its N-oxide) or quinolyl (or its N-oxide). A specific value for R3 is H. A specific value for Y is �S�, �O�, or �N(Ra)�. A more specific value for Y is �S�. A specific value for R4 is hydroxy, �N(Ra)(Rb), (C1-C6)alkyl, (C1-C6)alkoxy, or (C2-C7)alkanoyl. Another specific value for R4 is hydroxy, �N(Ra)(Rb), or (C1-C6)alkyl. A more specific value for R4 is �N(Ra)(Rb), or (C1-C4)alkyl. Another specific value for R4 is methyl, ethyl or propyl. A more specific value for R4 is methyl. Another specific value for R4 is �N(Ra)(Rb). Another specific value for R4is �N(CH2CH3)2. A specific value for �N(Ra)(Rb) is a 5-6 membered heterocyclic ring, optionally comprising 1, 2, or 3 N(Ra), non-peroxide O or S atoms. Another specific value for �N(Ra)(Rb) is pyrrolidino, piperidino or morpholino. A more specific value for �N(Ra)(Rb) is pyrrolidino. A specific compound of the invention is formula (IB):
where for R3 is H. The compounds of the invention, can be synthesized as illustrated in Scheme 1, starting with commercially available 2-aminobenzothiazoles. Alkylation of the 2-aminobenzothiazoles with various α-haloacetophenones provides the corresponding N-alkylated compounds, with alkylation occurring exclusively at the ring nitrogen. In protic solvents the ring alkylated fused thiazoles often formed aromatic ring-closed products with concomitant dehydration, leading to the corresponding closed ring derivatives. The corresponding IBT derivatives such as, for example, 3-15 can then be prepared by ring closure in refluxing ethanol or in methoxyethanol when higher temperatures were needed. Additional methods for preparing the compounds of the invention are described in Barchechath, S. et al., J Med Chem. 2005, 48, 6409-6422.
In cases where compounds are sufficiently basic or acidic to form stable nontoxic acid or base salts, administration of the compounds as salts may be appropriate. Examples of pharmaceutically acceptable salts are organic acid addition salts formed with acids which form a physiological acceptable anion, for example, tosylate, methanesulfonate, acetate, citrate, malonate, tartarate, succinate, benzoate, ascorbate, α-ketoglutarate, and α-glycerophosphate. Suitable inorganic salts may also be formed, including hydrochloride, sulfate, nitrate, bicarbonate, and carbonate salts. Pharmaceutically acceptable salts may be obtained using standard procedures well known in the art, for example, by reacting a sufficiently basic compound such as an amine with a suitable acid affording a physiologically acceptable anion. Alkali metal (for example, sodium, potassium or lithium) or alkaline earth metal (for example, calcium) salts of carboxylic acids can also be made. The compounds of formula (I) can be formulated as pharmaceutical compositions and administered to a mammalian host, such as a human cancer patient, in a variety of forms adapted to the chosen route of administration, i.e., orally or parenterally, by intravenous, intramuscular, topical or subcutaneous routes. Thus, the present compounds may be systemically administered, e.g., orally, in combination with a pharmaceutically acceptable vehicle such as an inert diluent or an assimilable edible carrier. They may be enclosed in hard or soft shell gelatin capsules, may be compressed into tablets, or may be incorporated directly with the food of the patient's diet. For oral therapeutic administration, the active compound may be combined with one or more excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. Such compositions and preparations should contain at least 0.1% of active compound. The percentage of the compositions and preparations may, of course, be varied and may conveniently be between about 2 to about 60% of the weight of a given unit dosage form. The amount of active compound in such therapeutically useful compositions is such that an effective dosage level will be obtained. The tablets, troches, pills, capsules, and the like may also contain the following: binders such as gum tragacanth, acacia, corn starch or gelatin; excipients such as dicalcium phosphate; a disintegrating agent such as corn starch, potato starch, alginic acid and the like; a lubricant such as magnesium stearate; and a sweetening agent such as sucrose, fructose, lactose or aspartame or a flavoring agent such as peppermint, oil of wintergreen, or cherry flavoring may be added. When the unit dosage form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier, such as a vegetable oil or a polyethylene glycol. Various other materials may be present as coatings or to otherwise modify the physical form of the solid unit dosage form. For instance, tablets, pills, or capsules may be coated with gelatin, wax, shellac or sugar and the like. A syrup or elixir may contain the active compound, sucrose or fructose as a sweetening agent, methyl and propylparabens as preservatives, a dye and flavoring such as cherry or orange flavor. Of course, any material used in preparing any unit dosage form should be pharmaceutically acceptable and substantially non-toxic in the amounts employed. In addition, the active compound may be incorporated into sustained-release preparations and devices. The active compound may also be administered intravenously or intraperitoneally by infusion or injection. Solutions of the active compound or its salts can be prepared in water, optionally mixed with a nontoxic surfactant. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, triacetin, and mixtures thereof and in oils. Under ordinary conditions of storage and use, these preparations contain a preservative to prevent the growth of microorganisms. The pharmaceutical dosage forms suitable for injection or infusion can include sterile aqueous solutions or dispersions or sterile powders comprising the active ingredient which are adapted for the extemporaneous preparation of sterile injectable or infusible solutions or dispersions, optionally encapsulated in liposomes. In all cases, the ultimate dosage form should be sterile, fluid and stable under the conditions of manufacture and storage. The liquid carrier or vehicle can be a solvent or liquid dispersion medium comprising, for example, water, ethanol, a polyol (for example, glycerol, propylene glycol, liquid polyethylene glycols, and the like), vegetable oils, nontoxic glycerol esters, and suitable mixtures thereof. The proper fluidity can be maintained, for example, by the formation of liposomes, by the maintenance of the required particle size in the case of dispersions or by the use of surfactants. The prevention of the action of microorganisms can be brought about by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases, it will be preferable to include isotonic agents, for example, sugars, buffers or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the compositions of agents delaying absorption, for example, aluminum monostearate and gelation. Sterile injectable solutions are prepared by incorporating the active compound in the required amount in the appropriate solvent with various of the other ingredients enumerated above, as required, followed by filter sterilization. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and the freeze drying techniques, which yield a powder of the active ingredient plus any additional desired ingredient present in the previously sterile-filtered solutions. For topical administration, the present compounds may be applied in pure form, i.e., when they are liquids. However, it will generally be desirable to administer them to the skin as compositions or formulations, in combination with a dermatologically acceptable carrier, which may be a solid or a liquid. Useful solid carriers include finely divided solids such as talc, clay, microcrystalline cellulose, silica, alumina and the like. Useful liquid carriers include water, alcohols or glycols or water-alcohol/glycol blends, in which the present compounds can be dissolved or dispersed at effective levels, optionally with the aid of non-toxic surfactants. Adjuvants such as fragrances and additional antimicrobial agents can be added to optimize the properties for a given use. The resultant liquid compositions can be applied from absorbent pads, used to impregnate bandages and other dressings, or sprayed onto the affected area using pump-type or aerosol sprayers. Thickeners such as synthetic polymers, fatty acids, fatty acid salts and esters, fatty alcohols, modified celluloses or modified mineral materials can also be employed with liquid carriers to form spreadable pastes, gels, ointments, soaps, and the like, for application directly to the skin of the user. Examples of useful dermatological compositions which can be used to deliver the compounds of formula (I) to the skin are known to the art; for example, see Jacquet et al. (U.S. Pat. No. 4,608,392), Geria (U.S. Pat. No. 4,992,478), Smith et al. (U.S. Pat. No. 4,559,157) and Wortzman (U.S. Pat. No. 4,820,508). Useful dosages of the compounds of formula (I) can be determined by comparing their in vitro activity, and in vivo activity in animal models. Methods for the extrapolation of effective dosages in mice, and other animals, to humans are known to the art; for example, see U.S. Pat. No. 4,938,949. Generally, the concentration of the compound(s) of formula (I) in a liquid composition, such as a lotion, will be from about 0.1-25 wt %, preferably from about 0.5-10 wt %. The concentration in a semi-solid or solid composition such as a gel or a powder will be about 0.1-5 wt %, preferably about 0.5-2.5 wt %. The amount of the compound, or an active salt or derivative thereof, required for use in treatment will vary not only with the particular salt selected but also with the route of administration, the nature of the condition being treated and the age and condition of the patient and will be ultimately at the discretion of the attendant physician or clinician. In general, however, a suitable dose will be in the range of from about 0.5 to about 100 mg/kg, e.g., from about 10 to about 75 mg/kg of body weight per day, such as 3 to about 50 mg per kilogram body weight of the recipient per day, preferably in the range of 6 to 90 mg/kg/day, most preferably in the range of 15 to 60 mg/kg/day. The compound is conveniently administered in unit dosage form, for example, containing 5 to 1000 mg, conveniently 10 to 750 mg, most conveniently, 50 to 500 mg of active ingredient per unit dosage form. Ideally, the active ingredient should be administered to achieve peak plasma concentrations of the active compound of from about 0.5 to about 75 μM, preferably, about 1 to 50 μM, most preferably, about 2 to about 30 μM. This may be achieved, for example, by the intravenous injection of a 0.05 to 5% solution of the active ingredient, optionally in saline, or orally administered as a bolus containing about 1-100 mg of the active ingredient. Desirable blood levels may be maintained by continuous infusion to provide about 0.01-5.0 mg/kg/hr or by intermittent infusions containing about 0.4-15 mg/kg of the active ingredient(s). The desired dose may conveniently be presented in a single dose or as divided doses administered at appropriate intervals, for example, as two, three, four or more sub-doses per day. The sub-dose itself may be further divided, e.g., into a number of discrete loosely spaced administrations, such as multiple inhalations from an insufflator or by application of a plurality of drops into the eye. The ability of a compound of the invention to act as a suppressor of p53 activity may be determined using pharmacological models which are well known to the art, e.g., as disclosed below. The invention will now be illustrated by the following non-limiting Examples. Biology. To examine the relative protective effect of the compounds a reliable and relatively rapid screening assay was required. We and others have found that the structural similarity of PFT-α and IBT with luciferin rendered luciferase-based reporter gene assays for apoptosis unreliable.19 Instead we assayed fluorescent dye retention described below in thymocytes after dexamethasone induced apoptosis. Dexamethasone (DEX) treatment of lymphocytes from the thymus initiates a signaling pathway that converges on the mitochondria-mediated molecular executioner cascade, leading to apoptosis within six hours.20 The six hour time point was chosen, as PFT-α was reported to inhibit DEX-induced apoptosis within this incubation period, and in our hands was found to be reliable and reproducible.5 All of the compounds were screened in the thymocyte assay for cytotoxicity and their ability to prevent dexamethasone-induced cell death. Murine thymocytes were exposed to 10 μM or 5 μM of each compound in the presence or absence of DEX for six hours and then assayed for their in vitro viability by 3,3′-dihexyloxacarbocyanine iodide (DiOC6) and propidium iodide (PI) staining, and flow cytometry. When cell death signals converge on the mitochondria, there is a loss of the inner mitochondrial membrane potential and cells can no longer retain the dye DiOC6 as an indicator of apoptosis. Once cells lose the integrity of the outer cell membrane, then the dye propidium iodide is no longer excluded from these cells. The percentage of viable cells, which retained DiOC6 and excluded PI was compared to cells treated with DEX alone. Compounds that significantly enhanced survival in repeated determinations (P≦0.01) were then selected for dose response study to determine the EC50 (Tables 1, 2, and
The effects of substitution at the para position of the phenyl ring of 1 and corresponding derivatives were investigated. A variety of substituents was selected on the basis of their electronic and lipophilic properties. These ability of these compounds to prevent cell death are shown in Table 3.
Chemistry. Melting points were obtained on a Mel-temp II capillary melting point apparatus and are uncorrected. Proton nuclear magnetic resonance spectra were obtained on a Varian Unity 500 at 499.8 MHz, or on a Varian Mercury at 400.06 MHz. The chemical shifts are expressed in δ values (parts per million) relative to tetramethylsilane (TMS) as internal standard. High resolution mass spectrometric analysis were performed on a Finnigan MAT900XP high resolution double focusing mass spectrometer using electron impact. Elemental analyses were performed by NuMega Resonance Labs, San Diego, Calif. Thin-layer chromatography was performed on silica gel 60 F-254 plates (EM Reagents). E Merck silica gel (230-400 mesh) was used for flash column chromatography. Biological Materials. Mice. C57B1/6 and p53−/− mice were purchased from The Jackson Laboratories (Bar Harbor, Me.). The mice were bred and maintained under standard conditions in the University of California, San Diego Animal Facility that is accredited by the American Association for Accreditation of Laboratory Animal Care. All animal protocols receive prior approval by the institutional review board. Reagents. Dexamethasone (DEX) was purchased from Sigma-Aldrich (St. Louis, Mo.). Other chemicals were purchased from Maybridge plc (Trevillett, Tintagel, Cornwall, UK), and Lancaster Synthesis (Windham, N.H.). Apoptosis assays. Thymocytes were harvested from young C57B1/6 mice and cultured at 37� C. in 5% CO2 in RPMI 1640 containing 10% FBS, 1% Penicillin/Streptomycin (Gibco BRL, Rockville, Md.). Thymocytes were plated at a density of 107 cells/mL and pre-incubated with 5-10 μM of each compound (from 10 mM stock in DMSO) for 30 minutes before induction of apoptosis. Apoptosis was induced with 5 μM dexamethasone or by exposure to 6 Gy gamma radiation. After 6 hours, cell apoptosis was assayed by propidium iodide (PI) and 3,3′ dihexyloxacarbocyanine iodide (DiOC6) staining. The cells were removed from the plate and incubated for 30 minutes in medium with 40 nM DiOC6 and 5 μg/mL PI and then analyzed by flow cytometry in a FACS caliber (Beckton-Dickinson, San Jose, Calif.). Viable cells had high DiOC6 (FL-1) and low PI (FL-3), whereas apoptotic cells had low DiOC6 (FL-1) and low PI (FL-3). To evaluate the EC50s the thymocytes were pre-exposed to graded concentrations of selected compounds for 30 minutes and then apoptosis was induced with 5 μM dexamethasone. After 6 hours the cells were harvested and stained as above. EC50 Determination The concentration (EC50) of each compound that inhibited dexamethasone-induced cell death by 50% was determined by nonlinear regression fitting of the data to a one-site model. Pseudo Hill slopes were determined by nonlinear regression fit of the data to a sigmoidal dose-response equation (variable slope): % viability=minimum % viability+(maximum−minimum % viability)/[1+10(log EC50−X)n], where X is the logarithm of inhibitor concentration, and n is the pseudo Hill slope and the maximum and minimum % viability were experimentally determined after dexamethasone exposure and drug treatment. EC50 values and 95% confidence intervals (CI) were derived from the sigmoid fits to the percent control transformed data shown using GraphPad Prism version 4.0b for Macintosh (GraphPad Software, San Diego, Calif.). Immunoblotting After removal of medium, cells were disrupted in lysis buffer (10 mM HEPES pH 7.9, 10 mM KCl, 0.1 mM EDTA, 0.1 mM EGTA, 1 mM dithiothreitol, 0.5 mM phenylmethylsulfonyl fluoride, and 0.6% NP-40) on ice. The nuclei were separated by centrifugation and the cytoplasmic sample removed. The nuclear pellets were resuspended in (20 mM HEPES pH 7.9, 0.4 M NaCl, 1.0 mM EDTA, 1.0 mM EGTA, 1 mM dithiothreitol, 0.1 mM phenylmethylsulfonyl fluoride) and the insoluble material was removed by centrifugation. The samples were heated to 70� C. for 10 minutes in loading buffer with 10 mM DTT. Each lane of an SDS-PAGE gel was loaded with 20 μg of protein. After electrophoresis, the proteins were transferred to a polyvinylidene difluoride (PVDF) membrane, blocked with 2% I-block� (Tropix Inc, Bedford, Mass.) containing 0.05% Tween-20 in PBS, and then incubated with anti-PUMA (AbCam, Inc., Cambridge, Mass.). Horseradish peroxidase-conjugated anti-IgG (Santa Cruz Laboratories, Santa Cruz, Calif.) was used as the secondary antibody. The membranes were developed using a chemiluminescence system (ECL detection reagent: Amersham Life Science, Aylesbury, UK). The membranes were reprobed with anti-actin (Sigma, St Louis, Mo.) to ensure equivalent loading. Synthesis Method A : General procedure for synthesis of PFT-α derivatives and aromatic analogs. Following a procedure reported by Singh et al., Indian J. Chem., Sect. B 1976, 14B, 997-998, a mixture of 2-amino-4,5,6,7-tetrahydrobenzothiazole (1 eq.) and the appropriate phenacyl bromide (1.1 eq.) was stirred in toluene at room temperature for 24 to 48 hours. The product crystallized out and was collected by filtration. The same procedure was used to synthesize aromatic analogs starting from commercially available of 2-aminobenzothiazole, some bearing a substituent on C-6. The aromatic analogs were made using the same procedures. General Procedure for Synthesis of Aromatic Analogs of IBT Derivatives. Method B: one-step ring closure. A mixture of 2-amino-benzothiazole (1 eq.) and the appropriate phenacyl bromide (1.1 eq.) was refluxed in ethanol for 90 minutes. The ring-closed product crystallized out upon cooling and was separated by filtration. Method C: two-step ring closure. 2-imino-3-phenacyl-benzothiaole hydrobromide or analog was refluxed in ethanol for 90 minutes. The ring-closed product crystallized out upon cooling and was separated by filtration. The title compound is prepared from α-bromo-4-(1-cyclopentyl)aceto-phenone (Hai et al., J. Org. Chem., 1958, 23, 39-42; Vejdelek et al., Collect. Czech. Chem. Commun. 1983, 48, 642-648) and 2-aminobenzothiazole using method A in 16% yield: mp 186-190� C. (dec.); 1H-NMR (DMSO-d6) 1.59 (m, 2H), 1.69, (m, 2H), 1.81 (m, 2H), 2.07 (m, 2H), 3.12 (m, 1H), 6.07 (s, 2H), 7.44 (d, 2H), 7.52 (m, 2H), 7.69 (t, 1H), 7.84 (t, 1H), 8.03 (d, 2H), 9.13 (s, 1H); HR-MS (FAB) m/z 337.1376 (M+); Anal. calculated for C20H21BrN2OS: C, 57.56; H, 5.07; N, 6.71, found; C, 50.82; H, 4.21; N, 7.65. The title compound is prepared from α-bromo-4-(1-pyrrolidino)acetophenone and 2-amino-4,5,6,7-tetrahydrobenzothiazole using method A in 43% yield: mp 285-290� C. (dec); 1H-NMR (DMSO-d6) 2.00 (s, 4H), 3.38 (m, 4H), 5.93 (s, 2H), 6.68 (d, 2H), 7.43 (t, 1H), 7.50 (t, 1H), 7.57 (d, 1H), 7.90 (d, 2H), 8.03 (d, 1H); Anal. C19H20BrN3OS (C, H, N). The title compound is prepared from 2-Bromo-1-furan-2-yl-ethanone and 2-aminobenzothiazole using method B in 30% yield: mp>250� C. (dec); 1H-NMR (DMSO-d6) 6.63 (s, 1H), 6.78 (s, 1H), 7.48 (t, 1H), 7.60 (t, 1H), 7.77 (s, 1H), 8.09 (d, 1H), 8.13 (d, 1H), 8.68 (s, 1H); Anal. (C13H9BrN2OS ⅔ H2O) C, H, N. A mixture of 9 (362 mg, 0.845 mmol) and copper (I) oxide (46 mg, 0.321 mmol) in a 30% aq. solution of NaOH (4 mL) was heated in a bomb for 4 hours. The mixture was neutralized and extracted with EtOAc. The solid recovered upon evaporation was purified by preparative TLC to give 4 in 47% yield: mp 275-277� C.; 1H-NMR (DMSO-d6) 2.31 (s, 3H), 6.95 (dd, 1H), 7.22 (d, 2H), 7.38 (d, 1H), 7.72 (d, 2H), 7.77 (d, 1H), 8.60 (s, 1H), 9.95 (s, 1H); HR-MS (EI) m/z 280.0662 (M+); Anal. calculated for C16H13BrN2OS � HBr: C, 59.90; H, 3.93; N, 8.73, found; C, 59.47; H, 4.43; N, 8.55. To a suspension of 13 (1.643 g, 4.76 mmol) in DMF (150 mL) at room temperature, was slowly added drop wise a solution of N-bromosuccinimide (933 mg, 5.24 mmol) in DMF (6 mL). The solvent was removed under high vacuum and the residue was slurried in ice water/ethanol (100 mL, 50%). A solid dropped out and was collected by filtration and rinsed with water to give 6 in 82% yield: mp>180� C. (dec); 1H-NMR (DMSO-d6) 2.36 (s, 3H), 7.31 (d, 2H), 7.51 (t, 1H), 7.61 (t, 1H), 7.90 (d, 2H), 8.11 (d, 1H), 8.44 (d, 1H); Anal. (C16H IBrN2S.⅓ H2O C, H, N. To a suspension of 7-Nitro-2-p-tolyl-imidazo[2,1-b]benzothiazole hydrobromide salt (128 mg, 0.328 mmol) in water at 80� C. were added 4 portions (10 eq.) of sodium dithionite over 30 minutes whereupon the suspension become clear and decolorized to give 7 in 22% yield: mp>210� C. (dec.); 1H-NMR (DMSO-d6) 2.32 (s, 3H), 6.80 (dd, 1H), 7.13 (d, 1H), 7.23 (d, 2H), 7.65 (d, 1H), 7.72 (d, 2H), 8.53 (s, 1H); HR-MS (EI) m/z 279.0821 (M+); Anal. calculated for C16H13N3S. 7/3 HBr: C, 41.05; H, 3.30; N, 8.98; found; C, 41.81; H, 2.86; N, 6.10. The title compound is prepared from 16 using method C in 65% yield: mp 0.250� C. (dec.); 1H-NMR (DMSO-d6) 1.57 (m, 2H), 1.66, (m, 2H), 1.79 (m, 2H), 2.03 (m, 2H), 3.00 (q, 1H), 7.33 (d, 2H), 7.45 (t, 1H), 7.59 (t, 1H), 7.78 (d, 2H), 8.01 (d, 1H), 8.06 (d, 1H), 8.77 (s, 1H); HR-MS (FAB) m/z 319.1267 (M+); Anal. calculated for C20H18N2S.� HBr: C, 70.93; H, 5.43; N, 8.27; found; C, 70.01; H, 5.82; N, 10.97. The title compound is prepared from 2-bromo-4′-methylacetophenone and 2-amino-6-bromobenzothiazole using method B in 24% yield: mp 205-208� C.; 1H-NMR (DMSO-d6) 2.31 (s, 3H), 7.23 (d, 2H), 7.73 (m, 3H), 7.91 (d, 1H), 8.32 (s, 1H), 8.70 (s, 1H); HR-MS (EI) m/z 341.9822 (M+). The title compound is prepared from 2-bromo-4′-methylacetophenone and 2-amino-6-bromobenzothiazole using method B in 62% yield: mp 274-276� C. (dec.); 1H-NMR (DMSO-d6) 2.33 (s, 3H), 3.84 (s, 3H), 7.18 (dd, 1H), 7.26 (d, 2H), 7.72 (m, 3H), 7.92 (d, 1H), 8.71 (s, 1H); Anal. (C17H15BrN2OS) C, H, N. The title compound is prepared from α-bromo-4-(1-pyrrolidino)acetophenone (Lancaster) and 2-aminobenzothiazole using method B and 10 hours of reflux in 16% yield: mp 283-287� C.; 1H-NMR (DMSO-d6) 1.98 (m, 4H), 3.31 (m, 4H), 6.68 (m, 2H), 7.51 (d, 1H), 7.66 (d, 3H), 8.11 (m, 2H), 8.71 (s, 1H); Anal. (C19H18BrN3S) C, H, N. A mixture of 2-aminobenzothiazole (2.116 g, 13.7 mmol), benzyl chloride (2.1 mL, 18.2 mmol) and sodium iodide (200 mg) were refluxed for 8 hours in methoxyethanol (25 mL). Crystallization occurred upon cooling. The crystals were filtered off and thoroughly washed with ether to give yellow crystals in 39% yield: mp 275-278� C.; 1H-NMR (DMSO-d6) 5.65 (s, 2H), 7.28, (d, 2H), 7.36 (m, 4H), 7.50 (t, 1H), 7.55 (d, 1H) 8.02 (s, 1H); HR-MS (EI) m/z 240.0719 (M+). A mixture of 2-aminobenzothiazole (2.104 g, 13.6 mmol) and 4-methylbenzyl chloride (2.25 mL, 12.5 mmol) was refluxed for 8 hours in methoxyethanol in the presence of a catalytic amount of sodium iodide (220 mg, 1.46 mmol). After cooling a precipitate formed which was filtered and rinsed with ether to give 35% yield: mp 243-248� C.; 1H-NMR (DMSO-d6) 2.26 (s, 3H), 5.65 (s, 2H), 7.20 (dd, 4H), 7.39 (t, 1H), 7.48 (t, 1H), 7.55 (d, 1H), 8.02 (d, 1H); HR-MS (EI) m/z 254.0870 (M+). To a solution of 2-aminobenzothiazole (1.018 g, 6.78 mmol) in dry pyridine (4 mL) was added portion wise, p-toluene sulfonyl chloride (1.42 g, 7.48 mmol). The solution turned yellow upon addition. After 5 minutes of stirring at rt, the mixture was heated (70-80� C.) for 5 minutes. The mixture was poured on a bed of ice and the resulting precipitate was filtered off and dried overnight in a dessicator under vacuum to give a yellow powder in 93% yield: mp 246-249� C.; 1H-NMR (DMSO-d6) 2.36 (s, 3H), 7.26, (dt, 1H), 7.30 (d, 1H), 7.37 (d, 2H), 7.40 (dt, 1H), 7.75 (d, 2H), 7.81 (d, 1H); Anal. (C14H12N2O2S2) C, H, N. To a solution of 20 (930 mg, 3.01 mmol) in DMF (15 mL) at room temperature was added NaH. After the effervescence subsided, 2-bromo-4′-methylacetophenone (775 mg, 3.36 mmol) was added. The reaction was first stirred at room temperature for an hour and then heated at 80� C. until completion (by TLC). The mixture was allowed to cool before being poured on ice (400 mL). A precipitate dropped out. It was filtered and washed with cold water and dried to give a pale yellow powder in quantitative yield: mp 180-182� C.; 1H-NMR (DMSO-d6) 2.35 (s, 3H), 2.43 (s, 3H), 5.86 (s, 2H), 7.33 (m, 3H), 7.42 (m, 3H), 7.57 (d, 1H), 7.64 (d, 2H), 7.91 (d, 1H), 7.97 (d, 2H); Anal. (C23H20N2O3S2) C, H, N. A mixture of 2-mercaptobenzothiazole (570 mg, 3.34 mmol) and 2-bromo-4′-methylacetophenone (870 mg, 3.67 mmol) was refluxed in ethanol (15 mL) for 90 minutes. The solution was cooled in the fridge to give a yellow solid in 18% yield: mp 76-78� C.; 1H-NMR (DMSO-d6) 2.42 (s, 3H), 5.15 (s, 2H), 7.36 (t, 1H), 7.41 (d, 2H), 7.44 (t, 1H), 7.78 (d, 1H), 8.00 (m, 3H); Anal. (C16H14BrNOS2� H2O) C, H, N. A mixture of 2-amino-4,5,6,7-tetrahydrobenzothiazole (2.296 g, 14.9 mmol) and ethyl bromoacetate (2.747 g, 16.4 mmol) was refluxed in ethanol (50 mL) containing 3 drops of Et3N for 90 minutes. The reaction mixture was concentrated to half volume in vacuo and the resulting precipitate was filtered and rinsed with cold ethanol, then ether to give a white powder in 79% yield: mp 222-224� C.; 1H-NMR (DMSO-d6) 1.25 (t, 3H), 1.74 (d, 4H), 2.39 (s, 2H), 2.52 (m, 2H), 4.22 (q, 2H), 4.95 (s, 2H), 9.67 (s, 2H); Anal. C11H17Br1N2O2S (C, H, N); HR-MS (EI) m/z 240.0931 (M+). The title compound is synthesized following the procedure described by Maeda et al. (Chem. Pharm. Bull. 1984, 32, 2536-2543). A solution of 4-Phenyl-1H-imidazole-2-thiol 24 (Maeda et al., Chem. Pharm. Bull. 1984, 32, 2536-2543) and 2-chlorocyclohexanone was refluxed in butanol for 3 hours. After cooling, the resulting precipitate was filtered to give 25 in 75% yield: mp 274-278� C.; 1H-NMR (DMSO-d6) 1.89 (m, 4H), 2.76 (m, 4H), 7.38 (t, 1H), 7.49 (t, 2H), 7.88 (d, 2H), 8.51 (s, 1H); HR-MS (EI) m/z 254.0874 (M+). The title compound is prepared from 2-bromo-4′-methylacetophenone and 2-aminothiazole using method A in 64% yield: mp>220� C. (dec); 1H-NMR (DMSO-d6) 2.42 (s, 3H) 5.79, (s, 2H), 7.06 (d, 1H), 7.35 (d, 1H), 7.44 (d, 2H), 7.92 (d, 2H), 9.55, (s, 2H); Anal. (C12H13BrN2OS ⅓ H2O) C, H, N. The title compound is prepared from 26 using a modification of method C with methoxyethanol instead of ethanol. The solvent was evaporated and the residue was recrystallized from ethanol in 49% yield: mp 260-262� C.; 1H-NMR (DMSO-d6) 2.35 (s, 3H), 7.31 (d, 2H), 7.55 (d, 1H), 7.72 (d, 2H), 8.16 (d, 1H), 8.41 (s, 1H); Anal. (C12H11BrN2S) C, H, N. To a solution of 2-hydroxybenzothiazole (1.17 g, 7.58 mmol) in THF (10 mL) was added Et3N (1.1 mL, 7.89 mmol). A solution of 2-bromo-4′-methylacetophenone (1.67 g, 7.06 mmol) in THf (10 mL) was added dropwise. Another equivalent of Et3N was added after 24 hours. Purification by flash chromatography on silicagel (CH2Cl2) gave a white powder in 67% yield: mp 168-170� C.; 1H-NMR (DMSO-d6) 2.43 (s, 3H), 5.62 (s, 2H), 7.21 (t, 1H), 7.27 (d, 1H) 7.32 (t, 1H), 7.42 (d, 2H), 7.70 (d, 1H), 8.01 (d, 2H); Anal. (C16H13NO2S) C, H, N. To a solution of 2-hydroxy-4,5,6,7-tetrahydrobenzothiazole (335 mg, 2.16 mmol) in DMF (3 mL) was added NaH (101 mg, 2.52 mmol). After stirring 10 minutes at room temperature, a solution of 2-bromo-4′-methylacetophenone (668 mg, 2.82 mmol) in DMF (5 mL) was added and the mixture was stirred overnight. Water was added and the mixture was extracted with EtOAc, dried over MgSO4 and the solvent was evaporated. The oil recovered was purified by flash chromatography on silicagel (CH2Cl2) to give 29 as a solid in 65% yield: mp 108-110� C.; 1H-NMR (DMSO-d6) 1.73 (s, 4H), 2.19 (s, 2H) 2.40 (m, 5H), 5.23 (s, 2H), 7.39 (d, 2H), 7.95 (d, 2H); Anal. (C16H17NO2S) C, H, N. A solution of quinazolin-4-yl-hydrazine (Gewald et al., J. Prakt. Chem./Chem.-Ztg. 1996, 338, 206-213) (265 mg, 1.65 mmol) was refluxed with tolualdehyde (400 μl, 3.39 mmol) in methanol for 1 hour. The solvent was evaporated and the resulting solid was triturated with water and filtered to give a yellow powder in 87% yield: mp 211-214� C.; 1H-NMR (DMSO-d6) 2.37 (s, 3H), 7.29 (d, 2H), 7.47 (dt, 2H), 7.67 (t, 1H), 7.88 (d, 3H), 8.20 (m, 1H), 8.51 (s, 1H), 11.65 (s, 1H); HR-MS (EI) m/z 262.1223 (M+). Four proapoptotic genes were tested by real time PCR. Bax, PUMA (p53-upregulated modulator of apoptosis) and noxa are transcriptional targets of p53, proapoptotic members of the Bcl-2 family required to initiate apoptosis, Villunger et al., Science 2003, 302, 1036-1038. PUMA and noxa belong to the BH3-only protein subclass while Bax is a multidomain member of the family. DR5 (also called killer) is an apoptosis-inducing membrane receptor for tumor necrosis factor-related apoptosis-inducing ligand (also called TRAIL and Apo2 ligand). DR5 is a transcriptional target of p53, and its over expression induces cell death in vitro (Finnberg et al., Molecular and Cellular Biology 2005, 25, 2000-2013). Upon treatment with DEX, all four genes were induced but only PUMA showed a significant increase of over 9-fold. Compound 15 was able to keep the level of the gene close to that of the control and even below it in the case of bax and killer ( Confirmation of the induction of PUMA following DEX treatment and ensuing reduction in the presence of PFT-α was made using western blot analysis. PUMA expression was almost nonexistent in the control sample but was induced by DEX. Addition of 1 and 15 reduced the amount of protein induction. PFT-α provided only partial inhibition of PUMA but 15 again proved to be more effective than PFT-α resulting in a total inhibition of PUMA expression in mouse thymocytes. ( The following illustrate representative pharmaceutical dosage forms, containing a compound of formula (I), for therapeutic or prophylactic use in humans.
The above formulations may be obtained by conventional procedures well known in the pharmaceutical art. All publications, patents, and patent documents are incorporated by reference herein, as though individually incorporated by reference. The invention has been described with reference to various specific and preferred embodiments and techniques. However, it should be understood that many variations and modifications may be made while remaining within the spirit and scope of the invention. Patent Citations
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Shelby County Schools board members got their first detailed look Wednesday, Feb. 19, at a set of more than two dozen changes in student attendance zones in the 2014-2015 school year that could affect nearly 7,000 students in the school system.
WASHINGTON (AP) – Superstorm Sandy will end up causing about $20 billion in property damages and $10 billion to $30 billion more in lost business, according to IHS Global Insight, a forecasting firm.
The Memphis Chapter of the International Association of Administrative Professionals will meet Monday, April 9, at 6 p.m. at the Hilton Memphis, 939 Ridge Lake Blvd. Judge Gina C. Higgins will speak about business law, its effect on business and the administrative assistant. Cost is $22. For reservations, contact Sharon Gardner at firstname.lastname@example.com or 752-6213.
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Hope and despair have co-existed for a long time along the stretch of Poplar Avenue between Danny Thomas Boulevard and Decatur Street. And for the past two years, the area has seen more change than just about any other inner-city avenue in Memphis.
The Memphis Regional Chamber's Metro Business Council will hold the annual Fall Sales & Marketing Summit today at 11:30 a.m. at Hilton Memphis, 939 Ridge Lake Blvd. The theme will be "Winning Strategies for Winning Clients" and will feature a panel of sales and marketing specialists. For more information, contact Angela Michaels at email@example.com or 543-3546.
The Advanced Distributed Learning Workforce Co-Lab at the University of Memphis hosts the first eLearning Summit Tuesday through Saturday at the U of M FedEx Institute of Technology, 365 Innovation Drive. The event features a series of three workshops that focus on various aspects of e-learning. Visit www.elearningsummit.org.

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Title: | Nike's Advertising History |
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Windows RT tablets are of questionable value when compared to the competition, and Acer can't resist the temptation to pile on. The PC maker has been making some interesting tablet and Ultrabook designs lately, but is choosing the wait and see approach for the ARM version of Windows 8.
"There are still users looking for more versatile products," said Acer President Jim Wong. He went on to add, "To be honest, there's no value doing the current version of RT." In regards to his opinion on Windows 8, Wong's opinion was mixed. "The launch was not ideal and the Surface tablet created some distraction, but Microsoft is now doing a good job of supporting its partners and explaining the touch-usage model to customers, Wong said."
When asked about the company's vision of the future, Wong clarified that a major focus will be placed on hybrids going forward. "We believe Windows 8 has some more chances. That's why we continue to invest in Windows 8 tablets," Wong said.
[1] http://www.maximumpc.com/user/justinkerr_0
[2] http://www.maximumpc.com/tags/windows_rt
[3] http://www.maximumpc.com/article/news/acer_claims_aspire_p3_cats_meow_ultrabooks2013
[4] http://www.citeworld.com/tablets/21824/acer-windows-rt-no-value
[5] http://www.twitter.com/justinkerr
[6] https://plus.google.com/u/0/102706118778464605651?rel=author
[7] http://www.maximumpc.com/tags/acer
[8] http://www.maximumpc.com/tags/hardware
[9] http://www.maximumpc.com/tags/operating_systems
[10] http://www.maximumpc.com/tags/software
[11] http://www.maximumpc.com/tags/windows
[12] http://www.maximumpc.com/tags/windows_8
[13] http://www.maximumpc.com/articles/news
[14] http://www.maximumpc.com/user/login?&commentfragment=comments_top_anchor


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You all remember when that cover that generated such controversy because of the character's unrealistic pose... Yeah they decided to fix it but the pose didn't change that much. At least its more pleasant to look at and more anatomically correct.
They fixed her cover...
It's only slightly better. There's still a lot wrong with it.
Unfortunately, they didn't fix what's inside. The issue is a trainwreck.
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Episode / Video Archive: Season 13
Videos are available for seasons 15 and later.
Michael Cleveland & Flamekeeper with Audie Blaylock (#1312)
Four-time winner of the IBMAs Fiddle Player of the Year award and Grammy-nominated musician Michael Cleveland brings dynamic traditional bluegrass to the stage with his band, Flamekeeper.
Premiered: Saturday, October 18, 2008 at 4:00/3:00 pm CT on KETKY
Berline-Crary-Hickman (#1311)
The Berline-Crary-Hickman trio features three of the most renowned bluegrass musicians to have taken up the form!
Premiered: Thursday night, March 18, 2010 at 12:00/11:00 pm CT on KETKY
Foghorn Stringband (#1310)
Foghorn Stringband delve into the traditional sounds of rural America, such as old-time, bluegrass, and other folk music.
Premiered: Saturday, July 12, 2008 at 4:00/3:00 pm CT on KETKY
Butch Waller & High Country (#1309)
Hailing from the West Coast, High Countrys repertoire offers a vital mix of powerful standards and original songs that focus on traditional bluegrass.
Premiered: Thursday night, July 3, 2008 at 12:00/11:00 pm CT on KETKY
The Lonesome River Band (#1308)
The Lonesome River Band brings its style of hard-driving, traditional bluegrass to the Jubilee stage.
Premiered: Sunday, February 21, 2010 at 2:00/1:00 am CT on KETKY
Hazel Dickens/Wade & Julia Mainer (#1307)
Three legendary performers--folk artist Hazel Dickens, and bluegrass pioneers Wade & Julia Mainer--offer up a masterful presentation of classic American music.
Premiered: Sunday, September 28, 2008 at 1:00/12:00 midnight CT on KETKY
Professors of Bluegrass (#1306)
The Professors of Bluegrass, a group of Yale University faculty members, perform their unique brand of traditional bluegrass.
Premiered: Saturday, June 14, 2008 at 11:00/10:00 pm CT on KET2
Timberline Drive/Jake Quesenberry & The MacRae Brothers (#1305)
Bowling Green's Timberline Drive and bluegrass legends Jake Quesenberry & The MacRae Brothers bring their skilled expertise to the Jubilee stage.
Premiered: Thursday night, October 6, 2011 at 12:00/11:00 pm CT on KETKY
The Farewell Drifters/Blue Moon Rising (#1304)
Two Tennessee bands perform classically inspired yet original bluegrass.
Premiered: Saturday, May 31, 2008 at 4:00/3:00 pm CT on KETKY
Dan Paisley and Southern Grass (#1303)
Dan Paisley & Southern Grass carry on the bluegrass tradition of the Paisley and Lundy families.
Premiered: Saturday, August 16, 2008 at 4:00/3:00 pm CT on KETKY
Adrienne Young & Little Sadie/Cadillac Sky (#1302)
Two groundbreaking groups--Adrienne Young & Little Sadie and Cadillac Sky--bring their unique brand of bluegrass to the Jubilee stage.
Premiered: Thursday night, May 15, 2008 at 12:00/11:00 pm CT on KETKY
The Grascals (#1301)
A band of rising stars displays its innovative mingling of bluegrass and country.
Premiered: Saturday, August 8, 2009 at 4:00/3:00 pm CT on KETKY


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Animals and LOVE - PhotosHi,Animals and LOVE - Photos2 years ago in Personal More Like This
thank you for so many notes with Valentine cards!
I wish you wonderful Valentine too and here is my series of "animals in love" photos
Wonderful leopards - PHOTOSHi!Wonderful leopards - PHOTOS3 years ago in Personal More Like This
Cats rulezz!
Huray, I am finally healthy. Finally it was not salmonellosis, but something very similar - so I took the same pills and now I am quite fine, which is good. Thanks for support
Is anybody here? + news + ZOO animals - PHOTOS.Hi!Is anybody here? + news + ZOO animals - PHOTOS.2 years ago in Personal More Like This
Ok, ok, ok, I am almost scared - what happaned? It seems that this site is totally empty for last week!
I wonder what caused this: Sandy? Or do Americans have some Halloween holiday so they are not online? Or... is there some secret and slow Zombie attack?
Of course dA decided to use this time to announce free shipping usually they do it before Christmas and I usually get pretty nice money from it. This time I got just few dolars, so I hope dA will repeat the action - just before X-mas as they did the last year. Because this site really seems to be empty now! as a response to a free shipping, I released Free calendar for 2013 - I think I have never had so few faves, views and downloads on it. What a pity, what a bad time
I know about Sandy, it was covered by Czech media a lot
Poor people, I hope everything will be fixed as soon as possible. I did not know it affected so many people
You also know I asked you to vote for Allerlei's photo - thank you
News + cute animals - PHOTOSHi!News + cute animals - PHOTOS3 years ago in Personal More Like This
I quit the job today.
I did not want to do it before finding another one, but I finally could not bear all that confused stuff and stress and low competence of the company. I was even supposed to help to spread something I considered to be evil for the world, so it was even more stressful.
Now I need to find a job as soon as possible. So I am poor again now XD
EDIT: people, you seem to be a bit confused - my job has never had anything to do with animals or photography! ZOO photography is just a hobby for me, I need to earn money to go to ZOOs. So my photography is not threatened by leaving the job, but it is threatened by having less money because of leaving the job if you know what I mean I just need to find another job
At least I finally have time to answer your notes and comments
Guilty and adorable baby animals - PHOTOSHi!Guilty and adorable baby animals - PHOTOS2 years ago in Personal More Like This
At first, thank you for very interesting discusion under my last journal - although I did not reply to every comment, be sure I read every single one! do not be afraid, I definitely do not judge USA and its citizens through some reality shows here in the Czech republic we also have really stupid shows and so, I think it is a world epidemy I always keep in mind that USA is huge country with a large scale of people of all the qualities some of you said they are ashamed to be Americans - I do not think there is a reason for it, as your country has f.e. many good universities or scientists!
I had a bad day today. I got totally stressed (I can not say why - maybe because of my stupid diploma thesis?) and when I get stressed, I eat. Holy mother, I ate such a crap today, that I feel really guilty and if I did not feel a strong resistance against it, I would like to kick myself to threw it all away I ate some caramel cream I prepared for a cake I made, then I ate TH
Very cute baby wild cats - PHOTOSHi,Very cute baby wild cats - PHOTOS3 years ago in Personal More Like This
I met my dear friend amrodel and we visited ZOO Jihlava together - it was full of baby cats!
Caracals, manuls and snow leopards had babies, so it was amazing to watch them - just the weather was bad, cloudy and even rainy, so the light made the photography difficult thanks for lovely meeting, Amrodel!
Here you can see some photos - I included also lovely Tamina the seal for Lunchi, of course
BABY LIONS - photos! But I am ill now :(Hi people!BABY LIONS - photos! But I am ill now :(3 years ago in Personal More Like This
Yesterday I went to ZOO Olomouc with my great friend amrodel - she has never been to this ZOO and because it is one of my the most favorite animal facilities, I really wanted to show her! And because amrodel is a lion lover and because baby lions were born in Olomouc, we could not have a better opportunity to meet! It is always a pleasure to see :devamrodel and we really had fun. It was fantastic to watch whole lion family - father imon was so desperate, because his babies wanted to play with his tail! He really seemed very unhappy about it. But we had fun.
Thank you for great meeting and for a great present (amrodel gave me a printed version of her famous and beautiful photo of my beloved wolf Lobo), friend
In the ZOO, I saw also something which took my breath away - I will show you later!
But after I got back, I got some leight gastric problems. I had them also today, but it was very far from anything serious. But during this eve
Tired + ZOO Prague, nice animals - PHOTOSHi!Tired + ZOO Prague, nice animals - PHOTOS3 years ago in Personal More Like This
It was my first weekend after a working week in my life - and I did not relax
I had to go to help my family, we painted a huge room in my grandfather's cottage (his free time house, where he loves to travel and work). I was driving and my father was not satisfied with it, which made me unhappy. I am really trying. But we have manual shift, of course, so I need to learn how to do it right... My father also wants me to drive faster than I think is safe. Do not get me wrong, I am not afraid of speed - if the road is good and quite empty, it is ok. But there was a narrow road with many bends, where bikers love to go and on the other hand, cars in opposite direction might have troubles to stay in their half. In such situation, I want to be more careful, but well...
I got even more angry about this, when we met a place of tragical car accident - I later found out that some young guy died there only hours ago. There were many cadles and young people staying close to the tree, crying
Relief, kiriban and ZOO Brno - brand new PHOTOSHi people!Relief, kiriban and ZOO Brno - brand new PHOTOS2 years ago in Personal More Like This
So my parents finally got back from Georgia. I have to say that I feel a great relief! Of course I knew local people are civilized that it is a nice country with fantastic nature, but it was still the Caucasus region and I definitely did not trust its roads and planes... My parents loved the trip and today and tomorrow we are looking at the photos they and their friends have taken. Nice to have them home
I also announce the newest kiriban as you did not catch the previos one: 904 444!
Catch my 904 444 pageviews and send me a printscreen via note as a proof to win a feature
(no feature for porn or animal cruelty, thanks for understanding)
Huh, I made a BIG mistake.
I read a part of Hunger games. I do not say it is a bad book - but I was shocked how bad I felt for that story! Among my friends, I am well-known for having no love for kids. But reading about deaths of all those children made me sick and although I stopped r
News+ ZOO photos + KIRIBAN is cloooose UPDATEHiNews+ ZOO photos + KIRIBAN is cloooose UPDATE2 years ago in Personal More Like This
Sorry for update, but it is time for Kiriban again - I annouce 899 999 this time
Catch my 899 999 pageviews and send me a printscreen as a proof via note to win a feature ;D
(no feature for porn or animal cruelty, thanks)
It is very close, so good luck
In case you noticed (I do not think so ), sorry for low activity in last days - I had too many things to do
I got back from Berlin, then we got prepared for a wedding! then my parents went to Georgia (I can not say I am happy about it), then it was the wedding, then it was my sister's birthday and then I went to Nuremberg - I got back today
I also answered all 120 notes, sorry it took so long! I already have 16 new ones...
Oh, a wedding!
Nope, it was NOT my wedding ;D it was a wedding of my beloved and admired cousin - I love him so much! He is so smart, honest, responsible, caring, intelligent, likeable, hansome, loving, ambicious, educated, helpful person and I can continue with superlatives. I lo
Baby polar foxes were born! + old PHOTOS of kitsHi!Baby polar foxes were born! + old PHOTOS of kits3 years ago in Personal More Like This
I just read the info, that baby polar foxes were born in local ZOO!
They are probably still too young to leave the burrow, but I might visit them soon to try to take some photos
Maybe it will kick me to buy a new year ticket - I have not visited the ZOO since my year ticket expired in March but yeah, I will buy it, I think. I definitely visit this ZOO more often than just six times a year
Some ooold photos of baby foxes (taken in 2009). Kits are so adorable
Big question for Americans + animals - PHOTOSHiBig question for Americans + animals - PHOTOS3 years ago in Personal More Like This
At first, there is a question I wanted to ask the Americans for years so I would be grateful for answer, thank you
I know it is completely silly to judge anything through movies, but...
Ok, the question of life and death:
Do you Americans REALLY enter your houses and flats with your shoes on?
I mean - in every movie or TV serie, people always get into their homes and walk through all the rooms with their shoes on. And there are so many scenes with teenagers jumping to their beds (!!!!!) right after getting back from school, still wearing shoes! it seems so silly to me - shoes are always the most dirty part of clothes, so I need to ask
For example in the Czech republic, it would be culturally unacceptable to enter the main rooms with shoes still on
The first place you see after you get into average Czech flat or house, is a small space where you take your shoes (and usually also coats, jumpers or jackets) off. And yes, Czech movies and series do
Thieves are annoying + animals - PHOTOS! + KIRIBANHi!Thieves are annoying + animals - PHOTOS! + KIRIBAN3 years ago in Personal More Like This
OMG, time for new kiriban again! Catch my 811 111 pageviews and send me a printscreen as a proof if you want to win a feature (no feature for porn or animal abuse, ok?)!
I can not believe that dA has not removed those stolen photos yet
I sent dA that "I swear to God, to Muhammad, to Buddha, to holy Cow, to the man on the moon, that I REALLY, but REALLY took those photos and I BEG YOU on my knees to remove the stolen ones" note, but nothing happened
I can not even post names of the thieves, because it would be me, who would have problems
I am just grateful that my nice friend TVD-Photography posted a journal (http://tvd-photography.deviantart.com/journal/Life-special-features-and-an-art-thief-294356735) about this problem, asking her watchers to report the thieves
Thanks, TVD-Photography, there is a tiny feature for you
EDIT - KIRIBAN IS CLOSE +fantastic animals - PHOTOHi!EDIT - KIRIBAN IS CLOSE +fantastic animals - PHOTO3 years ago in Personal More Like This
SORRY for update, but I would like to inform you, that kiriban is quite close usually it happens during night, so my American watchers have the biggest chance to catch it. Now it seems to be Europe's turn. Good luck!
Catch my 744 444 pageviews and send me a printscreen as a proof if you want to win a feature! (no feature for porn or animal cruelty, please )
I am really stressed now - I study so much but I do not remember anything
Those texts are so complicated that when I finish reading, I am not able to say what it was about
And those are some 6 000 pages!
I feel really uneasy about it. I have no time to learn that!
Also, this winter is so bad when it comes to snow
I am really happy I took many photos of wolves on snow last year, because it seems that winter 2011/12 will not offer any nice photo oportunity. There is NO snow here only on mountains, but not here. I love snow, so it makes me sad
Well, time for new Kiriban!
Catch my 744
Wonderful animals from ZOO Brno - PHOTOSHi,Wonderful animals from ZOO Brno - PHOTOS3 years ago in Personal More Like This
today I post some older photos taken by Canon Powershot S5 IS
Foxes, foxes, foxes - PHOTOS + kiribanHello,Foxes, foxes, foxes - PHOTOS + kiriban3 years ago in Personal More Like This
I announce the newest kiriban - it is 788 888 and it is quite close!
If you want to win a feature, send me a printscreen of it via note (no feature for porn or animal cruelty, sorry).
Gooooood luck!
Thank you for answering my questions about Austrian holidays
And the most important thing: my very nice and good friend BlueFox-cz has his birthday today! he is a very nice guy, real gentleman, who loves foxes and is active when it comes to animal rights
Blue fox, I wish you all the best, good health, loyal friends and many foxes around you!
Those fox photos are dedicated to you
News, thieves (yay) + ZOO animals - PHOTOSHelloNews, thieves (yay) + ZOO animals - PHOTOS3 years ago in Personal More Like This
I say that all the time - copyright problems come in waves XD at first, those were those two thieves (dA still has not removed the second stolen picture yet *facepalm*), then I found another three of my photos edited and posted by three different persons without any credit. No prob, all those people removed them or gave credit, when I contacted them. Yesterday, two different persons posted edited photos of my best friend Allerlei and today, I found another girl who manipulated few of my photos without any credit. I noticed her and I wait for her reply. And when you look at photography category, it is full of google photos again
EDIT: people, just a strange thing happened. I usually do not report people if they edited my work and posted it without my permission or without giving credit. I usually inform them and ask for credit. I wait even weeks or months for reply. But when I see such person has been warned several times and simply ignorre it, I do report.
Some annoying things, beautiful animals - PHOTOSHello!Some annoying things, beautiful animals - PHOTOS3 years ago in Personal More Like This
Mhm... I must say I am kind of annoyed - when I got to Austria, I saw that they have the same goods as here in the Czech republic - but the quality was much better. My parents agree with that - my mum even buys some chemicals for cleaning and washing powders in Austria, because the same things are much less effective here. If you buy the same chocolate here in the Czech republic and then you taste the same chocolate in Austria, the taste is very different. You might say: ,,ok, the Czechs probably prefer lower price to quality." But the most annoying thing is, that in the Czech republic, the same things have not only lower quality, but they have HIGHER price at the same time. And the month's salary of the Czechs is so much lower! it makes you really feel like some second-class person. Very stupid thing - experts have already proved that goods with MUCH lower quality are exported to countries of Central and Eastern Europe. But the European union said such praxis is OK, that
Wild, feroucious but beautiful - LEOPARDS - PHOTOSHello!Wild, feroucious but beautiful - LEOPARDS - PHOTOS3 years ago in Personal More Like This
Yay, I have only three last driving lessons before the final exam!
And I need to start writing my final thesis again! I bet you hate this line at least as much as me
I also made a totally delicious dessert with two different cremes, it took me at least 5 hours to finish it - but do not worry, one can eat only very small portion every day and I will give some pieces to my grandfather, friend and sister's boyfriend
And I need to ride a bike again - that Alpen trip interrupted my sporting again and I need to get used to it again
I at least had some nice hour of fast walking on Monday
Here come some cats
It is Sri Lankan leopard, Snow leopard, Amur leopard, Persian leopard, then again Sri Lankan leopard, Snow leopard, Amur leopard and Persian leopard. I hope you like it
Not so good news + animals from ZOO Brno - PHOTOSHi!Not so good news + animals from ZOO Brno - PHOTOS3 years ago in Personal More Like This
I am still very tired. I am really not used to waking up early, doing something for all 8 hours and then getting home late. I usually fall asleep very soon. I usualy wake up between 12 PM - 4 AM and I am able to just post one photo for dA. Or I wake up at 5 AM, which prevents me from doing anything - it is too late to exercise or to write something, but it is too soon to wake up. So I usually try to sleep. The worst thing is waking up at 6 AM - it is VERY early, but at the same time, it is not enough to fall asleep again. So as a result, it makes me very tired in the evening and this whole circle goes on and on.
I did not even have time to write any journal...
Maybe nobody noticed
Of course I did not ,,defend" my final thesis *facepalm* it is all my fault. I need to write it once again, I have the very last attempt to rewrite it *sad face*
I need to get used to my new regime, so I can exercise again.
I wanted to move at least through the weekend, but I spend it rather with gre
White wolves - PHOTOS (+ Kiriban VERY close)Hi!White wolves - PHOTOS (+ Kiriban VERY close)3 years ago in Personal More Like This
Kiriban (744 444) is very close!
And I am busy and totally stressed. I fell asleep - I wanted to sleep only some 10 minutes, but I woke up now, after some 6 hours! it makes me unhappy. I am afraid I will not make it, there are so many pages and I do not remember anything. But I want the graduation exams to be already behind me. My life style is just terrible now two more days and it would kill me
Super fresh PHOTOS from ZOO BRNOHello!Super fresh PHOTOS from ZOO BRNO3 years ago in Personal More Like This
My first visit in ZOO Brno in 2012 brought you those photos
*Losing weight news*
Oh my. I hate dead lines for studying. I have some 10 days to study for my graduation exams and I am so stressed! And I have a bad habit to eat a lot when I am stressed so I commited few really bad eating sins in last hours. And I had no time to exercise. So I am fixing it right now!
Sorry, funny thief + ZOO Brno - animal PHOTOSHi!Sorry, funny thief + ZOO Brno - animal PHOTOS3 years ago in Personal More Like This
At first, I want to apologize - because of my final exams, I could not answer notes, so finally I got 15 of them! All of them should be already answered, sorry for late replies. My brain was not able to work few days before and then also after graduation exams it should be ok now.
Also: sometime I might miss some of your comment's questions, sorry If you want to be 99,9% sure to get a reply from me, send me a note, please I always do my best to answer every single note. Very rarely, it can happen that I read some note minutes before leaving the flat, so I forget to answer it, but I think it is really VERY rare so once again - I love to answer questions, but if you want to be 99,9% sure to gain an answer, ask via note, please thank you!
At second - wohoo after pretty long time, my photo was reposted here on dA. It was not a big deal - I stayed calm and just told the person to remove the shot. She did remove it - but funny thing came, she posted a big h
White wolves and friends - PHOTOS and a bit sad.Hi!White wolves and friends - PHOTOS and a bit sad.3 years ago in Personal More Like This
Here are some new photos for you.
I am just sad when I look at Alex and Clair, the oldest females of the pack (the last photo). They both look so.... used do not be mistaken, it is not just winter/summer fur change. They are too slim, too used, losing too much fur, some parts of their bodies seem to be almost bare, they had even some small bleeding injuries... I think their bodies are too tired after so many pups it was nice to see them both meet next to the lake, as they walked away together as old friends, although they used to have at least one very big conflict which resulted in death of Alex's puppies. But it was a sad sight too. I do not want to be pesimistic, but I do not believe Alex and Clair will survive 2012 my personal guess is, that ZOO will put them to sleep in autumn.... but it is really my personal opinion
Nah, people: it is MY SPECULATION! ZOO has not announced anything like this, so do not blame it, please! I simply think that they would not survive wi


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recipe reviews for:
"These pickled eggs are marinated in a cider vinegar that is spiced with mustard and turmeric powder. The longer they set, the better they get."
Cooking Level: Intermediate
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Cooking Level: Expert
The best wintertime meals need a long, slow simmer while you're doing something else.
Our big collection of budget recipes will leave some jingle in your pocket.
Delicious recipes, party ideas, and cooking tips! Get a year of Allrecipes magazine for $7.99!
This simple Southern snack is a cinch to prepare.
See how to make classic pickled eggs.
Watch Chef John make quick-and-easy pickled peppers.


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Smoke bans move beyond passive steps
WITH the exception of a diehard few, not many people would willingly return to the days when smoking was commonplace in pubs, cinemas and even on flights.
WITH the exception of a diehard few, not many people would willingly return to the days when smoking was commonplace in pubs, cinemas and even on flights.
GUERNSEY'S ageing population is set to put extra strain on the purse-strings of the island's overspending health department, its deputy minister Martin Storey warned yesterday.
A CULTURE of health and safety in the workplace needs to come from the very top of the organisation, according to a leading figure in the industry.
PLANS to tackle high rates of teenage pregnancies will be addressed in a forthcoming sexual health strategy, a leading consultant has said.
NOW that the unredacted version of the Lightfoot review into the operation of the Ambulance and Rescue Service has finally been released, it is possible to see that withholding the information was secrecy for the sake of secrecy.
HEALTH and Social Services has finally committed to releasing full details of how much it costs to run the ambulance service.
A student who sat his GCSEs while suffering from his first bout of cancer has been recognised by a national charity.
I WAS horrified to read the Guernsey Press article on Judy Hayman, who may lose the vitamins she needs because the deputies are to decide on food supplements legislation, i.e., the EU law to control vitamins.
THE Health and Social Services board will discuss whether to lodge a second code of conduct complaint against its most vocal critic.
TO THE surprise of no one, HSSD survived this week's vote of no confidence by a hefty majority. It was not so much an endorsement of the brilliant job being done by the current board as an acknowledgement that the department would not benefit from more disruption. However, a bad-tempered debate raises doubts about what improvements islanders can hope for in their health service.
THE Health and Social Services Department wishes to correct statements made in relation to the evaluation and recommended next steps for the bowel cancer screening programme on pages one and two of the Guernsey Press on Monday 27 January.
HEALTH and Social Services is not guaranteeing value for money, a review by the Public Accounts Committee has concluded.
FLOODING at the offices of Environmental Health and Pollution Regulation caused one of its employees to collapse after an allergic reaction, a tribunal has heard.
HEALTH's new board has 'failed dismally' on its election promises, a deputy who tabled a vote of no confidence in its predecessor has said.
A WARD closed as a 'temporary' measure in the wake of Health and Social Services £2.5m. overspend will reopen nearly a year after operations were cancelled.
PATIENTS might have to wait an extra week for their injectable drugs after an unexpected fungal mould was found in the hospital's pharmacy department.


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Klime Kovaceski, chef-proprietor of Crystal Café, dismisses those kinds of guilty pangs, both from a business side and a consumer point of view. A Macedonian, he has lived with unsettled violence as a fact of life. "I grew up with the fighting of wars," he recollects. "It's pretty scary, and it obviously affects everyone. But going to restaurants and ordering a cognac and talking for hours is all part of the [wartime] fabric." He notes that his clientele hasn't really fallen off; rather people seem to be dining out as an act of confirmation of life or rebellion against the loss of it.
So while things may not seem so rosy -- or rosé -- at the moment, it is within striking distance that South Beach will manage to emerge victorious from the slump. Of course we must do our part as locals and show some loyalty to our hometown eateries. As a bonus you can also help out by exhibiting an appetite. On October 11 a national campaign will mark the inauguration of the Window of Hope Family Relief Fund, which will aid those who lost relatives who worked in Windows on the World and other culinary support systems of the World Trade Center.
35 NE 40th St.
Miami, FL 33137
Category: Restaurant > American
Region: Midtown/Wynwood/Design District
433 Washington Ave.
Miami Beach, FL 33139
Category: Restaurant > Italian
Region: South Beach
Restaurants, which will include Pacific Time (restaurateurs interested in participating are invited to contact Eismann), will donate monies made on this night, the one-month anniversary of the catastrophe, to the fund. And it may indeed be self-serving to consider helping the victims' families in this way. But no doubt Eismann's pan-Eastern fare is a reward far more singular than the Oreo and juice you get for donating blood, which is never going to make it out of South Florida anyway.


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Member Since 20 Jan 2012Offline Last Active Jan 24 2015 08:52 PM
Profile Feed
So Sorry I missed the Art Journal Chat too. Where do I go to see the schedule of the Art Journal Chats?
Apr 27 2014 07:20 AM
Cheryllann I'm sending you a PM, there is no actual schedule, but usually it's once a month. :)
Apr 27 2014 08:05 AM
teecee → cherryyllaann
Congratulations on your win. It's a beautiful page! http://scrapgirls.com/blog/
Mar 29 2014 08:36 AM
Belle → cherryyllaann
Congrats on your winning layout in the blog this week. Beautiful layout.
Mar 29 2014 06:03 AM
MariJ → cherryyllaann
Hi CheryllAnn! Just wanted to remind you about the Art Journaling Chat & Challenge tomorrow, Monday March 17th at 9PM ET. Hope to see you there! :)
Mar 16 2014 12:30 PM
Any suggestions for how to get a sock out of a vacuum cleaner hose? That's apparently what happens when the kids help clean?!...
Mar 01 2014 05:49 PM
my thought too Belle.. I used one on my shower drain the other week... don't ask!
Mar 02 2014 07:47 AM -
Wire Hanger - Ahhh, I had not considered that. Definitely keep in mind for the future.
Mar 02 2014 09:09 AM
Fished the 5K Turkey Trot with Jaxon, Spinach Quiche in the oven and prepping Turkey and sides. Having a great day and hoping all of you are too. Happy Thanksgiving.
Nov 28 2013 10:47 AM
Its official, kids home with strep throat. Jax seems to like chicken soup broth and the flat ginger ale is working. Dylan is not quite as bad off. Hopefully we'll get some sleep tonight.. Till then I'm sorting through challenges to catch up on.
Nov 25 2013 01:48 PM
Great suggestion Andrea, never heard of that. Poor little guys, we are in summer cold/flu season here so I feel your pain. Hugs to them and you!
Nov 25 2013 07:20 PM -
Florida granny
I hope they feel better soon. Meanwhile, it's good to have some scrapping time.
Nov 26 2013 07:56 AM
Becster → cherryyllaann
Thanks so much for commenting on my heritage LO! The letter you mentioned is part of the CRO_FeelingThankful collection!
Nov 16 2013 12:26 PM
This morning we brought in Jaxon's candy bags with Poem/Layout attached. Gave one to his principal. She loved it so much she had Jaxon read his poem to the school - on camera- during morning announcements. Luckily I had my camera with me. So unexpected and appreciated. So happpy for Jaxon :)
Oct 31 2013 10:30 AM
here is the link to his poem/layout if you want to see it. http://scrapgirls.net/forum/gallery/image/146645-oct-30-hnc-witches-and-pet-dragons/
Oct 31 2013 10:31 AM


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India's budget: Saturday morning pictures
Business and finance
Greece's election: A crowded field
Syriza edges closer to victory, but with uncertainty over its coalition partner4
Greek politics: Remaking the political landscape
A new burst of optimism about the economy is not yet luring many voters back to the two mainstream parties11
Corruption in Greece: Political minefield
A police investigation finds evidence that public money was misused10
Greek taxation: Doubts over competence
Not a single prominent tax evader has yet been convicted23
New biography: For Greece a tear
The political drama of the Papandreous12
Politics in Greece: Election games
The manoeuvring ahead of a new vote12
Greece's dysfunctional politics: Tomorrow and tomorrow
How not to form a government52
Charlemagne: Europe against the people?
Efforts to save the euro cannot run against the will of the voters indefinitely245
Greece's politicians: In their own time
Greece will get its coalition government. But that comes with its own dangers7
Greece and the euro: Papandreou's people
The uncertain political ramifications surrounding Greece and the euro36
The G20 and the euro's crisis: The burning fuse
The G20 summit is dominated by the crisis in Greece and the euro zone68
The euro crisis: The Greeks will vote on something, and other euro news
Fun times in Athens, and Frankfurt5
A euro referendum: Greece's woes
The markets are not the euro's only threat. Voters may be too195
Charlemagne: A Greek bearing gifts
George Papandreou chose the wrong time to call for a referendum91
Financial markets: Greece lightning
The prime minister's botched referendum plan has left the debt deal in trouble28
The euro-zone crisis: A Greek drama
George Papandreou, the Greek prime minister, seems to be on the way out, but that will not solve the country's problems269
Greece's woes: Debts, downturns and demonstrations
How much more can Greeks take?71
Alternative indicators: Behind the bald figures
Receding hairlines and other signals of where the economy is heading3
Greece's finances: Out of the frying pan
Greece has gained some breathing space, for now0
EU summit: Greek opposition under EU fire
European leaders turn their fire on the Greek opposition leader53
Greece and the euro: The brewing storm
George Papandreou's new government has passed its first pressing test. But disaster may still loom88
Greece's crisis: Papandreou survives round one
The new Greek government wins a vote of confidence. But there are choppy waters ahead51
Charlemagne: It's all Greek to them
The streets of Athens are in the grip of an ugly mood322
Greece's economic woes: The labours of austerity
The international plan to rescue Greece is instead starting to paralyse it20
Greece's budgetary woes: A long odyssey
Support for the government's austerity programme is still holding up, just13
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Toll sticker 2015: Azure is the color of the year
Important: the new vignette in azure is valid from December 1, 2014 - the "old" vignette in lime but can still easily be used until 31 January 2015. This means that from February 1, 2015 applies only to new vignette in Azure.
Another innovation: according to law, the price of the vignette on the harmonized index of consumer prices (HICP) adapted - and by 2.1 percent. Should read: A car-year vignette costs EUR 84.40 - EUR 33,60 for motorcycles are due.
Rates valid as December 1st 2014
10-day toll sticker | 2-month toll sticker | Annual toll sticker | |
Motorbike | 5,00 | 12,70 | 33,60 |
Car (vehicle up to and including 3.5 tonnes MPW) | 8,70 | 25,30 | 84,40 |
Authentication features
All Austrian types of toll sticker are backed with a two-dimensional hologram. This authentication feature is used as the main protection against counterfeit products. When tilting the vignette both the federal eagle and the motorway symbol is visible in the silver shimmering field. The number "14" refers to the yearly period of validity - in this example from 1st December 2013 to 31st January 2015. The black-colored cross with the words "Abziehen und aufkleben" ("stripping and stick") on the liner of the vignette will continue to maintain a multiple use of the vignette is excluded.
For further information about the vignette please see the outgoing FAQ Link.
Rates valid as December 1st 2013
The toll sticker 2014 is lime colored. Prices for 2014 has been adjusted at the annual harmonized index of consumer prices (HICP) in accordance with the legal requirements and increased by 2.6 percent. The toll sticker (vehicles with a maximum permissible gross weight of up to 3.5 tonnes) entitled to use the Austrian motorways and expressways. The requirement applies to cars, motorcycles and light motor homes.
The use of Austria's motorways and expressways is subject to charge. All vehicles up to 3.5 tonnes maximum permissible weight (MPW) - i.e. cars, motorbikes and small motor homes - need a valid motorway toll sticker. Depending on the type of toll sticker.
10-day toll sticker | 2-month toll sticker | Annual toll sticker | |
Motorbike | 4,90 | 12,40 | 32,90 |
Car (vehicle up to and including 3.5 tonnes MPW) | 8,50 | 24,80 | 82,70 |
With around 6,000 points of sale in Austria and abroad, ASFINAG ensures the smooth sale of toll stickers. The toll sticker is easy to affix to the vehicle. Compared with the toll stickers of other countries, the prices of the 'pickerl', or motorway 'pickerl' as it is called locally, are moderate, which has made a decisive contribution to acceptance of the toll system in Austria.
The prices of the toll stickers vary depending on the type of vehicle and the period of validity.
In connection with the correct application and removal of the toll sticker the same questions appear again and again. What is essential to ensure the purchase and the proper handling of the vignette explains our short video with Conny in a very clear and entertaining way.
Drive safely, stick the toll sticker!
As one hundred percent user-financed company thus ASFINAG receives any financial assistance from the state budget. All proceeds from sale vignette, truck tolls and special toll roads will only be reinvested in the construction, operation and safety of the primary road network in Austria.


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"Can I see another's woe, / And not be in sorrow too?"
William Blake
"Can I see another's woe, and not be in sorrow, too? Can I see another's grief, and not seek for kind relief?"
"Who hath sorrow? Who hath woe?/ They who do not answer no;/ They whose feet to sin incline/ While they tarry at the wine."
Carry Nation
"Sainthood emerges when you can listen to someone's tale of woe and not respond with a description of your own."
Andrew V. Mason
"Not so, not so, no load of woeNeed bring despairing frown;For while we bear it, we can bear,Past that, we lay it down."
Sarah Williams
"The hues of bliss more brightly glow, Chastis'd by sabler tints not of woe"
Thomas Gray
"Woe to him that buildeth a town with blood, and stablisheth a city by iniquity! / Behold, is it not of the LORD of hosts that the people shall labour in the very fire, and the people shall weary themselves for very vanity? / For the earth shall be filled with the knowledge of the glory of the LORD, as the waters cover the sea."


text:
What does CWAG stand for?
What does CWAG mean? This page is about the various possible meanings of the acronym, abbreviation, shorthand or slang term: CWAG.
We've found a total of 6 definitions for CWAG:
Sort by:PopularityAlphabeticallyCategory
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CWAG | Citizens Water Advocacy Group | ||||
CWAG | Conference of Western Attorneys General | ||||
CWAG | Canadian Women and Geography | ||||
CWAG | Canberra Women's Adventure Group | ||||
CWAG | Coalition of Wisconsin Aging Groups | ||||
CWAG | Central Waterfront Advisory Group |
Still can't find the acronym definition you were looking for? Use our Power Search technology to look for more unique definitions from across the web!
"CWAG." Abbreviations.com. STANDS4 LLC, 2015. Web. 27 Feb. 2015. <http://www.abbreviations.com/CWAG>.


text:
Italy > Lombardy > Ponte Caffaro > Bastei
Bastei rankings
Difficulty ranking world: 300 (all)
Ranking Italy: 64 (all)
Ranking Alps: 107 (all)
Difficulty score: 152.55 what?
Bastei ratings
(rate it)
The Bastei is situated in Lombardy
and belongs to the
Starting from Ponte Caffaro,
the Bastei ascent is 11 km long. Over this distance, you climb 1129 heightmeters.
The average percentage thus is 10.3 %.
Look for other sides to climb the Bastei.
Find all information on climbs and cycling in Lombardy and find all climbs in Lombardy on a map. |
Other climbs close to the Bastei: within 10 km - within 20 km - within 50 km |
No stories submitted yet. Did you climb this mountain?
Did you climb the Bastei? Please send your cycling story, description or comments. You can also send us video's or send us pictures on the Bastei.
© climbbybike™ All rights reserved 2005 -


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