text:
The present invention relates to compositions and methods for inhibiting unwanted angiogenesis, particularly those of ocular tissues. The treatment, inhibition, and/or prevention of choroidal neovasculature (CNV) is provided, along with an animal model for CNV and imaging techniques that permit the screening of potential agents as anti-angiogenesis and anti-CNV agents.
1. A method for treating a human having the wet form of age-related macular degeneration, the method comprising ophthalmically administering to the human a composition comprising an effective amount of rapamycin to treat the age-related macular degeneration, wherein the rapamycin is dissolved in polyethylene glycol, and wherein the composition is administered by placement of the composition into the vitreous or by placement of the composition between the conjunctiva and the sclera of an eye of the human.
2. The method of
3. The method of
4. The method of
5. The method of
6. The method of
7. A method for inhibiting the transition in a human from the dry form of age-related macular degeneration to the wet form of age-related macular degeneration, the method comprising ophthalmically administering to a human having the dry form of age-related macular degeneration a composition comprising an effective amount of rapamycin to inhibit the transition to the wet form of age-related macular degeneration, wherein the rapamycin is dissolved in polyethylene glycol, and wherein the composition is administered by placement of the composition into the vitreous or by placement of the composition between the conjunctiva and the sclera of an eye of the human.
8. The method of
9. The method of
10. The method of
11. The method of
12. A method for treating an angiogenesis-mediated disease or condition of the retina or choroid in a mammal, the method comprising ophthalmically administering to the mammal an effective amount of a composition comprising rapamycin and a second component comprising polyethylene glycol, wherein the composition is suitable for ophthalmic administration by injection and the composition is administered by placement of the composition into the vitreous or by placement of the composition between the conjunctiva and the sclera of an eye of the mammal.
13. The method of
14. The method of
15. The method of
16. The method of
17. The method of
18. The method of
19. The method of
20. The method of
21. The method of
22. A method for treating an angiogenesis-mediated disease or condition of the retina or choroid in a mammal, the method comprising ophthalmically administering to the mammal an effective amount of a composition comprising polyethylene glycol and an agent selected from the group consisting of rapamycin, tacrolimus, everolimus, pimecrolimus, and temsirolimus, wherein the composition is suitable for ophthalmic administration by injection and the composition is administered by placement of the composition into the vitreous or by placement of the composition between the conjunctiva and the sclera of an eye of the mammal.
23. The method of
24. The method of
25. The method of
26. The method of
27. The method of
28. The method of
29. The method of
This application claims benefit of priority to U.S. Provisional Patent Application 60/412,088, filed Sep. 18, 2002, which is hereby incorporated in its entirety as if fully set forth.
This invention was supported in part by Grant Nos. Y12727 from the U.S. National Institutes of Health. The Government may have certain rights in this invention.
The present invention relates to compositions and methods for inhibiting unwanted angiogenesis, including that of ocular tissues. In particular, compositions and methods for the treatment of choroidal neovasculature (CNV) in ocular diseases are provided. While the invention is exemplified with respect to rapamycin (sirolimus) and tacrolimus, the invention provides for the use of the family of "limus" compounds to inhibit unwanted angiogenesis.
The retina of the eye contains the cones and rods that detect colors. In the center of the retina is the macula lutea, which is about ⅓ to � cm in diameter. The macula provides detailed vision, particularly in the center (the fovea), because the cones are higher in density. Blood vessels, ganglion cells, inner nuclear layer and cells, and the plexiform layers are all displaced to one side (rather than resting above the cones), thereby allowing light a more direct path to the cones.
Under the retina are the choroid, comprising a collection of blood vessels embedded within a fibrous tissue, and the deeply pigmented epithelium, which overlays the choroid layer. The choroidal blood vessels provide nutrition to the retina (particularly its visual cells).
There are a variety of retinal disorders, whose current treatment is not optimal. The retina may tear, form holes and separate from the underlying choroid.
Age-related macular degeneration (AMD) is the major cause of severe visual loss in the United States for individuals over the age of 55. AMD occurs in either an atrophic or (less commonly) an exudative form. In exudative AMD, blood vessels grow from the choriocapillaris through defects in Bruch's membrane, and in some cases the underlying retinal pigment epithelium (choroidal neovascularization or angiogenesis). Organization of serous or hemorrhagic exudates escaping from these vessels results in fibrovascular scarring of the macular region with attendant degeneration of the neuroretina, detachment and tears of the retinal pigment epithelium, vitreous hemorrhage and permanent loss of central vision. This process is responsible for more than 80% of cases of significant visual loss in patients with AMD.
Several studies have recently described the use of laser photocoagulation in the treatment of initial or recurrent neovascular lesions associated with AMD (Macular Photocoagulation Study Groups (1991) in Arch. Ophthal. 109:1220; Arch. Ophthal. 109:1232; Arch Ophthal. 109:1242). Unfortunately, AMD patients with subfoveal lesions subjected to laser treatment experienced a rather precipitous reduction in visual acuity (mean 3 lines) at 3 months follow-up. Moreover, at two years post-treatment treated eyes had only marginally better visual acuity than their untreated counterparts (means of 20/320 and 20/400, respectively). Another drawback of the procedure is that vision after surgery is immediately worse.
Choroidal neovascularization (CNV) has proven recalcitrant to treatment in most cases. Laser treatment can ablate CNV and help to preserve vision in selected cases not involving the center of the retina, but this is limited to only about 10% of the cases. There is no other treatment available to correct CNV. Unfortunately, even with successful laser photocoagulation, the neovascularization recurs in about 50-70% of eyes (50% over 3 years and >60% at 5 years). (Macular Photocoagulation Study Group, Arch. Ophthalmol. 204:694-701 (1986)). In addition, many patients who develop CNV are not good candidates for laser therapy because the CNV is too large for laser treatment, or the location cannot be determined so that the physician cannot accurately aim the laser. Thus, until the present invention, there has been a long-felt need for methods that will prevent or significantly inhibit choroidal neovascularization.
In addition to AMD, choroidal neovascularization is caused by such retinal disorders as: presumed ocular histoplasmosis syndrome, myopic degeneration, angioid streaks and ocular trauma. Angiogenic damage associated with retinal and intravitreal neovascularization occurs in a wide range of disorders including diabetic retinopathy, venous occlusions, sickle cell retinopathy, retinopathy of prematurity, retinal detachment, ocular ischemia and trauma.
Citation of the above documents is not intended as an admission that any of the foregoing is pertinent prior art. All statements as to the date or representation as to the contents of these documents is based on the information available to the applicant and does not constitute any admission as to the correctness of the dates or contents of these documents.
The present invention provides compositions and methods that are effective in inhibiting unwanted angiogenesis, specifically choroidal neovascularization (CNV) that is associated with ocular diseases such as age related macular degeneration (AMD) and histoplasmosis syndrome. Compositions of the invention for inhibiting angiogenesis comprise active agents in the "limus" family of compounds, which bind to members of the immunophilin family of cellular proteins, including cyclophilins and FK506-binding proteins (FKBPs), to inhibit angiogenesis in choroidal membranes. Non-limiting members of the "limus" family of compounds include sirolimus (rapamycin) and its water soluble analog SDZ-RAD, tacrolimus, everolimus, pimecrolimus, CCI-779 (Wyeth), AP23841 (Ariad), and ABT-578 (Abbott Laboratories) as well as analogs and derivatives thereof.
A therapeutic amount of the active agents of the invention may be administered to a patient by a variety of different routes and can be given in dosages that are safe and provide angiogenic inhibition at internal sites. The present invention thus provides methods of treating mammalian diseases characterized by undesired and uncontrolled angiogenesis by administering a composition comprising one or more active agents of the invention. In a particular embodiment of the invention, methods to inhibit or treat choroidal neovascularization (CNV) of the eye are provided.
Thus, the present invention is especially useful for treating certain ocular neovascular diseases such as macular degeneration, including age-related macular degeneration (AMD). The invention is particularly useful in the treatment or inhibition of the wet form of AMD wherein blood vessels grow from their normal location in the choroids into an undesirable position under the retina. Leakage and bleeding from these new blood vessels results in vision loss and possibly blindness. The invention also provides methods for inhibiting the transition from the dry form of AMD (wherein the retinal pigment epithelium or RPE degenerates and leads to photoreceptor cell death and the formation of yellow deposits called drusen under the retina) to the wet form of AMD. The invention thus also provides methods for the treatment of the dry form of AMD.
Compounds which are contemplated for use in the present invention are administered to the patient to halt the progression of the disease and permit reductions in, or regression of, the neovascularization. Other diseases that can be treated using the present invention include, but are not limited to, diabetic retinopathy, neovascular glaucoma and retrolental fibroplasia.
Accordingly, and in a first aspect, the invention provides compounds, compositions, kits and methods to inhibit unwanted angiogenesis as well as neovascularization in the retina of a human or animal. In a second aspect, the invention provides a treatment for diseases mediated by angiogenesis or choroidal neovascularization in a subject. In a further aspect, the invention provides methods for preventing, inhibiting or treating the wet form of AMD, including inhibiting the loss of vision associated therewith.
Another aspect of the invention is the use of the above described methods in combination with other methods known for the treatment of angiogenesis, neovascularization, and the wet form of AMD as well as reducing the loss of visual acuity associated therewith. Moreover, the invention provides methods for the visualization of blood vessels by use of lipophilic dyes in a body as well as an animal model for choroidal neovascularization that can be applied as an assay for rapid identification of additional anti-angiogenic, anti-neovascularization, and anti-AMD compounds.
Advantages and novel features of the invention will be set forth in part in the description, examples and figures which follow, all of which are intended to be for illustrative purposes only, and not intended in any way to limit the invention, and in part will become apparent to those skilled in the art on examination of the following, or may be learned by practice of the invention.
The invention provides methods for the treatment of disorders involving angiogenesis and neovascularization, including ophthalmic disorders, and in particular retinal disorders involving macular degeneration, choroidal neovascularization, and the like in the retina or between the retina and its underlying choroidal tissue, or involving choroidal tissue, as described above. The methods comprise a novel use for the immune suppressor, rapamycin, which is also known as macrocyclic lactone sirolimus (commercially available as Rapamune�, Wyeth-Ayerst) (See, Physician's Desk Reference, 55th edition). According to the Merck Index, 12th edition, Raparnune is also known as RAPA, RPM, sirolimus, AY22989, and NSC-226080.
Although Sirolimus is known as an immunosuppressant, it has been reported as an anti-angiogenic compound in the context of primary and metastatic tumors (Guba et al., Nature Medicine 18(2):128-135 (February 2002) and Guba et al., Chir. Forum Exp. Klin. Forsch. Band 30, pages 37-39 (2001)). In those studies, there is no discussion related to other types of neovascularization, such as choroidal neovascularization. Instead, there is a discussion of the involvement of vascular endothelial growth factor (VEGF) and the serum levels thereof VEGF is a factor implicated in numerous indications without certainty that therapies directed thereto are efficacious in treating said indications. For example, VEGF has been suggested as involved in the formation of pathogenic new vessels in AMD, although VEGF activity has never been tested in an animal model with regard to AMD. Therefore, the ability to treat AMD by targeting VEGF activity remains experimental.
In addition to the use of sirolimus, the invention provides for the use of other immunophilin binding compounds as well as rapamycin derivatives and analogs to treat angiogenesis and neovascularization. Non-limiting examples of such compounds include SDZ-RAD, tacrolimus, everolimus, pimecrolimus, CCI-779 (Wyeth), AP23841 (Ariad), and ABT-578 (Abbott Laboratories) as well as those described in U.S. Pat. Nos. 5,527,907; 6,376,517; and 6,329,386. Additional derivatives include those disclosed in published U.S. Patent application 2002/0123505. All of these documents are hereby incorporated by reference as if fully set forth.
The invention also provides for the use of the above agents in combination with other agents and therapies for the treatment of angiogenesis or neovascularization, particularly CNV. Non-limiting examples of such additional agents and therapies include pyrrolidine, dithiocarbamate (NFκB inhibitor); squalamine; TPN 470 analogue and fumagillin; PKC (protein kinase C) inhibitors; Tie-1 and Tie-2 kinase inhibitors; inhibitors of VEGF receptor kinase; proteosome inhibitors such as Velcade™ (bortezomib, for injection; ranibuzumab (Lucentis™) and other antibodies directed to the same target; pegaptanib (Macugen™); vitronectin receptor antagonists, such as cyclic peptide antagonists of vitronectin receptor-type integrins; α-v/β-3 integrin antagonists; α-v/β-1 integrin antagonists; thiazolidinediones such as rosiglitazone or troglitazone; interferon, including γ-interferon or interferon targeted to CNV by use of dextran and metal coordination; pigment epithelium derived factor (PEDF); endostatin; angiostatin; anecortave acetate; acetonide; triamcinolone; tetrathiomolybdate; RNA silencing or RNA interference (RNAi) of angiogenic factors, including ribozymes that target VEGF expression; Accutane™ (13-cis retinoic acid); ACE inhibitors such as quinopril or perindozril; inhibitors of mTOR (mammalian target of rapamycin); 3-aminothalidomide; pentoxifylline; 2-methoxyestradiol; colchicines; AMG-1470; cyclooxygenase inhibitors such as nepafenac, rofecoxib, and diclofenac; t-RNA synthase modulator; metalloprotease 13 inhibitor; acetylcholinesterase inhibitor; potassium channel blockers; endorepellin; purine analog of 6-thioguanine; cyclic peroxide ANO-2; (recombinant) arginine deiminase; epigallocatechin-3-gallate; cerivastatin; analogues of suramin; VEGF trap molecules; Visudyne™ and other photosensitizers with photodynamic therapy (PDT); and laser photocoagulation.
"Macular degeneration" is characterized by the excessive buildup of fibrovascular deposits in or beneath the macula and retina and the atrophy and/or dislodgement of the retinal pigment epithelium (RPE). The administration of rapamycin appears to limit excessive angiogenesis, such as choroidal neovascularization in age-related macular degeneration (AMD), which may occur without such treatment. As used herein, the term "angiogenesis" means the generation of new blood vessels ("neovascularization") into a tissue or organ. An "angiogenesis-mediated disease or condition" of the eye or retina is one in which new blood vessels are generated in a pathogenic manner in the eye or retina, resulting in loss of vision or other problem, e.g., choroidal neovascularization associated with AMD.
The methods of the invention include a preferred embodiment using rapamycin and or other active agents in vitro or in vivo. When administered in vitro, the method is used, for example, to screen for, or assay the effects of, additional candidate active agents for the activity of controlling or reducing neovascularization or angiogenesis in retinal or choroidal tissue or cells. This may be used as a helpful assay for additional anti-angiogenesis or CNV agents. When administered in vivo the method is used, for example, to treat a patient having a predisposition to develop the choroidal neovascularization typically seen in AMD, or to prevent or inhibit choroidal neovascularization in such a patient, or to reduce choroidal neovascularization in an AMD patient. Prevent, inhibit and reduce are given their ordinary meanings with regard to the effect of the active agents of the invention on choroidal neovascularization. A patient having a predisposition or in need of prevention may be identified by the skilled practitioner by established methods and criteria in the field. The skilled practitioner may also readily diagnose individuals as in need of inhibition or treatment based upon established criteria in the field for identifying unwanted angiogenesis and/or neovascularization.
An effective amount of the drug is that amount which provides the therapeutic effect sought, e.g., an therapeutically effective dose of rapamycin or drug equivalent would be the amount which reduces choroidal neovascularization in an AMD patient, or which inhibits or completely prevents choroidal neovascularization in a patient predisposed to AMD or who, even without predisposition, shows early signs of AMD. Thus, the therapeutically effective dose may not be the same in every patient treated with rapamycin. An effective amount also refers to the amount of drug which inhibits angiogenesis or neovascularization in a model or assay therefor, such as that disclosed by the present invention.
"Patient" preferably refers to a subject who has, or who may develop, choroidal neovascularization associated with exudative AMD unless treated by the preferred methods of the present invention. Such a patient is preferably a mammal, more preferably a human, although the present methods are also application to model experimental animals and veterinary animal subjects.
An active agent of the invention, such as rapamycin, is preferably administered orally, intravenously, topically, intraocularly, intramuscularly, locally or in an ocular device. More preferably the mode of administration is selected from the following: intraocular injection, subretinal injection, subscleral injection, intrachoroidal injection, subconjunctival injection, topical administration, oral administration and parenteral administration. Most preferably the active agent is directly administered to the retinal area by subretinal injection, although less invasive modes of administration may be developed that are equally as effective. Formulations for timed release or delayed release over time are also provided by the present invention.
The dosage of the active agent will depend on the condition being treated, the particular agent, and other clinical factors such as weight and condition of the human or animal and the route of administration of the agent. It is to be understood that the present invention has application for both human and veterinary use. For administration to humans, an effective dosage is one that inhibits choroidal neovascularization. In the case of rapamycin, an inhibiting amount that can be employed ranges generally between about 0.1 to 300 mg/kg/day, preferably between approximately 0.5 and 50 mg/kg/day, and most preferably between approximately 1 to 10 mg/kg/day. Dosages of various agents of the invention for treating various conditions can be refined by the use of clinical trials on the present invention. Additionally, dose ranges for the practice of the invention include those disclosed in U.S. Pat. Nos. 6,376,517 and 5,387,589, which are hereby incorporated by reference as if fully set forth.
An active agent of the invention, such as rapamycin, may be subjected to conventional pharmaceutical operations, such as sterilization and/or may contain conventional adjuvants, such as preservatives, stabilizers, wetting agents, emulsifiers, buffers etc. The agents may also be formulated with pharmaceutically acceptable excipients for clinical use to produce a pharmaceutical composition. Formulations of the present invention suitable for oral administration may be presented as discrete units such as capsules, cachets or tablets each containing a predetermined amount of the active ingredient; as a powder or granules; as a solution or a suspension in an aqueous liquid or a non-aqueous liquid; or as an oil-in-water liquid emulsion or a water-in-oil emulsion and as a bolus, etc. Stated differently, the active agents of the invention may be used to prepare a medicament for the treatment of any of the conditions described herein.
For administration, an active agent such as rapamycin may be combined with one or more adjuvants appropriate for the indicated route of administration. The active agent may be admixed with lactose, sucrose, starch powder, cellulose esters of alkanoic acids, stearic acid, talc, magnesium stearate, magnesium oxide, sodium and calcium salts of phosphoric and sulphuric acids, acacia, gelatin, sodium alginate, polyvinylpyrrolidine, and/or polyvinyl alcohol, and tableted or encapsulated for conventional administration. Alternatively, the compounds of this invention may be dissolved in polyethylene glycol, propylene glycol, carboxymethyl cellulose colloidal solutions, ethanol, corn oil, peanut oil, cottonseed oil, sesame oil, tragacanth gum, and/or various buffers. Other adjuvants and modes of administration are well known in the pharmaceutical art and may be used in the practice of the invention. The carrier or diluent may include time delay material, such as glyceryl monostearate or glyceryl distearate alone or with a wax, or other materials well known in the art.
The formulations of the invention include those suitable for oral, ophthalmic, (including intravitreal or intracameral) nasal, topical (including buccal and sublingual), or parenteral (including subcutaneous, intramuscular, intravenous, intradermal, intratracheal, and epidural) administration. The formulations may conveniently be presented in unit dosage form and may be prepared by conventional pharmaceutical techniques. Such techniques include the step of bringing into association the active ingredient and the pharmaceutical carrier(s) or excipient(s). In general, the formulations are prepared by uniformly and intimately bringing into associate the active ingredient with liquid carriers or finely divided solid carriers or both, and then, if necessary, shaping the product.
Further provided in another aspect of the invention is a pharmaceutical composition comprising: a therapeutically effective amount of an active agent of the invention, such as rapamycin, and a pharmaceutically acceptable carrier suitable for administration to the eye or eye tissue.
In addition, a kit is provided, comprising at least one vial comprising a therapeutically effective amount of an active agent of the invention, such as rapamycin, and a second vial comprising a pharmaceutically acceptable carrier suitable for administration to the eye or eye tissue. Other kits of the invention comprise components such as the active agent of the invention for use in the practice of the methods disclosed herein, wherein containers, each with one or more of the various reagents (typically in concentrated form) utilized in the methods, including, for example, buffers and other reagents as necessary, are also included. A label or indicator describing, or a set of instructions for use of, kit components in a method of the present invention, will also be typically included, where the instructions may be associated with a package insert and/or the packaging of the kit or the components thereof.
Diseases associated with retinal/choroidal neovascularization that can be treated according to the present invention include, but are not limited to, diabetic retinopathy, macular degeneration, retinopathy of prematurity, infections causing a retinitis or choroiditis, presumed ocular histoplasmosis, myopic degeneration, angioid streaks, ocular trauma, and AMD. Other non-limiting examples of diseases and unwanted conditions that may be treated with the present invention include, but are not limited to, pseudoxanthoma elasticum, vein occlusion, artery occlusion, carotid obstructive disease, Sickle Cell anemia, Eales disease, myopia, chronic retinal detachment, hyperviscosity syndromes, toxoplasmosis, trauma and post-laser complications. Other diseases include, but are not limited to, diseases associated with rubeosis (neovasculariation of the angle) and diseases caused by the abnormal proliferation of fibrovascular or fibrous tissue, including all forms of proliferative vitreoretinopathy, whether or not associated with diabetes.
In addition to the compositions and methods for the treatment of unwanted conditions, the invention also provides methods for the visualization of blood vessels in a body. Such methods may also be viewed as methods of detectably labeling blood vessels in a body for subsequent visualization. Conventional ways to process tissue samples for blood vessel visualization are both labor intensive and time consuming. To improve the efficiency in tissue preparation, a technique called Vessel Painting is provided by the present invention. The basic concept of Vessel Painting is to selectively stain the inner lining of blood vessels with fluorescent dye. Blood vessels are lined with endothelial cell membrane, a lipid bilayer membrane that can be stained directly with a lipophilic dye. The key to this technique is a specially formulated solution, the Vessel Paint, which contains a lipophilic dye. Non-limiting examples of such dyes are available from Molecular Probes, and they include DiI, DiO, DiO, DiD, DiA, and DiR, which are long-chain dialkylcarbocyanines and dialkylaminostyryl dyes used as neuronal tracers. By intracardiac perfusion of an animal with Vessel Paint followed by a wash, optionally in a fixative solution such as, but not limited to 4% paraformaldehyde solution, blood vessels are stained instantaneously. Tissues can be viewed by fluorescence microscopy immediately after perfusion. The staining is remarkably bright with very low background so that high contrast images can be obtained using objective lenses of different magnification powers.
The invention also provides for an animal model of ocular neovascularization. The model is discussed in detail below, but generally it is based upon the injection of material into the subretinal space of an animal's eye. Any suitable non-human animal model for ocular disease may be used, and the injected material may range from Matrigel™, an extract of extracellular matrix (ECM) proteins from the murine EHS (Engelbreth-Holm-Swarm) tumor that is widely used as reconstituted basement membrane in cell culture experiments, to a simple solution of rat tail collagen I, bovine collagen I, and human collagen I (such as that available from BD Biosciences). See Gautreau, A. et. al., PNAS, 96: 7300. (1999); Abir R. et al. Hum Reprod, 14:299. (1999); and Abir R. et al. Fertil Steril, 75:141. (2001). Other sources of collagen may be used, including those produced by using lyophilized collagen (such as rat tail collagen from Roche) dissolved in 0.1�DME pH 4.0 (DME power from Life technologies without NaHCO3, make a 10� solution but with the pH color indicator, and use HCl to adjust pH to 4.0, then dilute this solution with water to make the 0.1�DME solution) and a 10% acetic acid solution.
Without being bound by theory, and offered to improve the understanding of the invention, the injection of a collagen (or protein) solution is believed to be sufficient to mimic the abnormal deposits that occur in AMD after injection into the subretinal space of rats to induce new blood vessels invasion. Such animal models may be advantageously used to screen candidate active agents of the invention for activity against angiogenesis, neovascularization (such as CNV), and AMD. Non-limiting examples of such methods include those comprising the administration (by any method disclosed herein) of a candidate agent to said animal and determining the effect (increase, decrease, or no change) on angiogenesis or neovascularization in said animal.
The following examples are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the present invention, and are not intended to limit the scope of what the inventors regard as their invention nor are they intended to represent that the experiments below are all and only experiments performed. Efforts have been made to ensure accuracy with respect to numbers used (e.g. amounts, temperature, etc.) but some experimental errors and deviations should be accounted for. Unless indicated otherwise, parts are parts by weight, molecular weight is weight average molecular weight, temperature is in degrees Celsius, and pressure is at or near atmospheric.
Within this application, unless otherwise stated, the techniques utilized may be found in any of several well-known references, such as: Molecular Cloning: A Laboratory Manual (Sambrook et al., Cold Spring Harbor Laboratory Press (1989); "Guide to Protein Purification" in Methods in Enzymology (M. P. Deutshcer, ed., (1990) Academic Press, Inc.); Culture of Animal Cells: A Manual of Basic Technique, 2nd Ed., Liss, Inc., New York, N.Y., (1987).
In search of effective treatments for CNV, a simple animal model was created by injecting 2-3 μl Matrigel™ to the subretinal space of adult Sprague-Dawley rats using a 33-guage needle connected to a 10 μl Hamilton microsyringe. A week or more later the animals were sacrificed by CO2 inhalation and perfused with Vessel Paint, a new visualization technique recently developed in the inventors' lab that enables screening agents, including chemical compounds and proteins, for their potential in inhibiting CNV (Wen, ARVO Abstract, March 2002). Vessel Painting comprises the use of a solution containing DiI, followed by 4% paraformaldehyde solution, and is discussed further below.
The anterior portion of the eye, including the cornea and lens, was removed, and the eyecup was imbedded in 5% agarose. Serial thick sections (100 μm) of the eye were cut on a vibratome and mounted on glass slides. Eye sections were examined by fluorescence microscopy for neovascularization in the Matrigel™ deposit. Serial optical sections were obtained using confocal microscopy. Three-dimensional reconstruction of newly developed blood vessels was achieved using Auto Visualiz-3D (Autouant Imaging, Inc.). Protein leakage of the new vessels was detected by assessing color change in the Matrigel™ deposit after intravenous injection of Evan's Blue dye.
Neovascularization was observed as early as 7 days after Matrigel™ injection, and extensive CNV was evident 10 days after Matrigel™ injection in all of the eyes injected. New blood vessels, originated from choriocapillaris exclusively, invaded the Matrigel™ deposit and formed extensive networks 14 days after Matrigel™ injection. Three-dimensional reconstruction clearly showed that the new vessels originated from the choroid. The Matrigel™ deposit became light blue in color after Evan's blue injection, as compared with pale white surrounding tissue, indicating the lack of barrier. Discform scar was observed 30 days after Matrigel™ injection.
Thus, the subretinal Matrigel™ deposits induce CNV in the subretinal space, mimicking the pathology seen in exudative, or wet form, AMD, and thereby providing an improved animal model for researching the pathology of CNV and for testing potential therapies.
In the initial characterization of the model, it was suspected that there was a possible involvement of an inflammatory reaction to Matrigel™ in the generation of neovascularization. As a result, two known immunosuppressants were tested, cyclosporin and raparnycin.
Oral administration of cyclosporin (15 mg/kg/d, given 4 day before Matrigel™ injection thorough 10 days after injection) had no effect on CNV. In marked contrast, however, oral rapamycin (Rapamune�, 1.5 mg/kg/d, given 4 days before Matrigel™ injection thorough 10 day after injection) resulted in complete inhibition of CNV development in the 16 eyes tested. Rapamycin is commercially available as oral solution, marketed as Rapamune Oral Solution by Wyeth-Ayerst. Thus, the anti-CNV properties of rapamycin by local administration were further examined.
Since rapamycin is not soluble in water, it was either dissolved in DMSO (tested in 8 eyes) or suspended in PBS (tested in 6 eyes), then mixed with Matrigel™. The mixed Matrigel™-containing rapamycin was injected to the subretinal space. At a dose of 25 μg/μl rapamycin (or 30 μg/injection, since each injection used 1.2 μl of Matrigel™), there was again a complete inhibition of CNV in rapamycin-treated eyes.
In further experiments, and using the methods described above, the amount of rapamycin was reduced to 2.5 μg/l. At this amount (the word "concentration" is not being used since rapamycin is not soluble in water), the crystals of rapamycin were clearly visible, even after 10 days. In each case, there was no detectable neovascularization in treated eyes. Concerns with insolubility, if they exist, may be addressed by the use of soluble active agents of the invention, such as, but not limited to, SDZ-RAD.
Rapamycin thus has the potential to inhibit or prevent CNV in human patients. In addition, local administration of rapamycin is evidently a practical approach to CNV treatment, which is particularly advantageous given the potential disadvantages effects of systemic administration.
The retina from a normal 3 months old Sprague-Dawley rat, sacrificed by CO2 overdose was perfused with Vessel Paint (DiI, 0.1 mg/ml), followed by 4% paraformaldehyde. The retina was dissected and postfixed in the same fixative for 1 hr, rinsed in phosphate buffered saline (PBS), and flat-mounted on glass slides. Microphotographs were taken on a Nikon E800 microscope.
Microphotographs of the flat-mount retina showed that vessels stain brightly with a low background. Endothelial cell nuclei were also stained and easily identifiable. The spatial structure of the vasculature was well preserved and the deep retinal capillaries were also visible.
The vascular network is better appreciated by images to show its three-dimensionality. Excellent 3-D images of vasculature can be obtained using the corrosion-casting technique and scanning electron microscopy (SEM), see Konerding M A (1991) Scanning electron microscopy of corrosion casting in medicine. Scanning Microsc. 5:851-865. However, corrosion casting is technically challenging and time-consuming, as is SEM. Alternatively, 3-D images can be reconstructed from a stack of serial optical sections by confocal microscopy. Bright staining and a high signal-to-noise ratio by Vessel Painting make it possible to obtain serial optical sections of samples as thick as 100-150 μm by laser scanning confocal microscopy without significant deterioration of signal from the bottom of the sample even using objective lenses of low magnification power. High quality of 3-D images at different viewing angles can be reconstructed from a stack of 2-D digital images using commercially available software.
Matrigel™ is an extract of extracellular matrix (ECM) proteins from the murine EHS (Engelbreth-Holm-Swarm) tumor and is widely used as reconstituted basement membrane in cell culture experiments. It is also used to assess angiogenic or antiangiogenic agents in an in vivo assay, the Matrigel™ plug assay (Passaniti A, Taylor R M, Pili R, et al. (1992) A simple, quantitative method for assessing angiogenesis and antiangiogenic agents using reconstituted basement membrane, heparin, and fibroblast growth factor. Lab Invest. 67:519-528). Pathological studies indicate an association between CNV and abnormal deposits of extracellular matrix (ECM) in the location between the retinal pigment epithelium (RPE) and Bruch's membrane. In order to mimic the abnormal deposits that occur in AMD, Matrigel™ was injected into the subretinal space of rats. New blood vessels invade Matrigel™ deposits shortly after Matrigel™ injection.
In the following, Matrigel™ (1.2 μl) was introduced to the subretinal space by injection into eyes of Sprague-Dawley rats. At a given time after injection, the animal was sacrificed and perfused with Vessel Paint, followed by 4% paraformaldehyde solution. The anterior portion of the eye was removed and the eyecup was then embedded in 5% agarose. Serial cross sections (100 μm thick) were cut on a vibratome.
New blood vessels originating from the choriocapillaris were detectable as early as 4 days after Matrigel™ injection and became well developed by 10 days after injection. A DIC image of a cross section of an eye, from a 2 month old animal 10 days after Matrigel™ injection, is shown in
The permeability of newly formed blood vessels was assessed by Evans Blue assay. Evans Blue (60 mg/kg in PBS) was injected intravenously to a Sprague-Dawley rat whose eyes had been injected with Matrigel™ 10 days before. A choroid-retina preparation was dissected, flat-mounted and viewed by fluorescence microscopy. Evans Blue staining was only seen in the Matrigel™ injected area, indicating the leaky nature of the new vessels.
In initial screens of potential anti angiogenic agents using the Matrigel™ model, rapamycin demonstrated a remarkable ability to inhibit new blood vessel formation. Rapamycin, clinically used as immunosuppressant (Kahan B D (2001) Sirolimus: a comprehensive review. Expert Opin. Pharmacother. 2:1903-1917), binds to a FKBP (FK506 binding protein) to form an FKBP-rapamycin complex, which in turn inhibits the function of mTOR (mammalian target of rapamycin), a central controller of cell growth (Schmelzle T, Hall M N (2000) TOR, a central controller of cell growth. Cell 103:253-262). Rapamycin inhibits endothelial cell proliferation (Vinals F, Chambard J C, Pouyssegur J (1999) p70 S6 kinase-mediated protein synthesis is a critical step for vascular endothelial cell proliferation. J. Biol. Chem. 274:26776-26782) and the response to VEGF (Yu Y, Sato J D (1999) MAP kinases, phosphatidylinositol 3-kinase, and p70 S6 kinase mediate the mitogenic response of human endothelial cells to vascular endothelial growth factor. J. Cell Physiol. 178:235-246) and bFGF (basic fibroblast growth factor) and PDGF (platelet-derived growth factor). See Cao et al. (1995) Effects of rapamycin on growth factor-stimulated vascular smooth muscle cell DNA synthesis. Inhibition of basic fibroblast growth factor and platelet-derived growth factor action and antagonism of rapamycin by FK506. Transplantation 59(3):390-5 and Ruygrok et al. (2003) Rapamycin and cardiovascular medicine. Intern. Med. J 33(3):103-9. It has been shown to block tumor angiogenesis (Guba M, von Breitenbuch P, Steinbauer M, et al. (2002) Rapamycin inhibits primary and metastatic tumor growth by antiangiogenesis: involvement of vascular endothelial growth factor. Nat. Med. 8:128-135).
Adult Sprague-Dawley rats (n=22) were injected with Matrigel™ to the subretinal space. Animals were fed with rapamycin once a day at a dose of 3 mg/kg started 4 days before Matrigel™ injection. Eyes were collected at 10 days (n=18) or 20 days (n=4) after Matrigel™ injection and processed as described in Example 4. Tissue sections were examined by fluorescence microscopy. None of the eyes collected at 10 days contained any new blood vessels invading the subretinal space in rapamycin treated animals. In the 20 days group, some newly formed blood vessels started to invade the Matrigel™ area. The amount of CNV in this group was semiquantified as (+). In comparison, CNV in control animals (10 days) were graded as (+++˜++++), as shown in
In another group of animals, rapamycin was mixed with Matrigel™ (suspension, 1 μg/μl, n=6; 10 μg/μl, n=11) and co-injected to the subretinal space with Matrigel™ (in-gel delivery). Eyes were collected 10 days after injection and processed as described in Example 4. No CNV was found in any of the eyes treated with rapamycin by in-gel delivery.
In all experiments, rapamycin had no discernable effect on the normal vasculature of the eye. Cyclosporin, also an immunosuppressant, failed to inhibit CNV formation either when administrated orally (100 mg/kg/d, n=3) or in-gel (25 μg/μl, n=3) by the same experimental paradigm.
FK506, which is not water soluble, was mixed with Matrigel™ (as a suspension) at 10 μg/μl, and 1.2 μl were injected into the subretinal space as described above. Eyes were collected 10 days after injection and blood vessels were stained with Vessel Paint. Eyes were embedded in 5% agarose and serial sections were cut (100 μm thick) on a vibratome. CNV was examined by fluorescence microscopy and CNV index of each eye was calculated. Results are shown below.
Therefore, FK506 inhibited CNV by 60%.
All references cited herein, including patents, patent applications, and publications, are hereby incorporated by reference in their entireties, whether previously specifically incorporated or not.
Having now fully described this invention, it will be appreciated by those skilled in the art that the same can be performed within a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the invention and without undue experimentation.
While this invention has been described in connection with specific embodiments thereof, will be understood that it is capable of further modifications. This application is intended to cover any variations, uses, or adaptations of the invention following, in general, the principles of the invention and including such departures from the present disclosure as come within known or customary practice within the art to which the invention pertains and as may be applied to the essential features hereinbefore set forth.
Patent Citations
Non-Patent Citations
Referenced by
Legal Events
Original Image


text:
Dana Morgan sliced some berries and put them on a plate you could sit on a quarter, with room to spare.
The artist baked the dish at a very high temperature, but it came out looking field-fresh. Yet you eat this fruit plate only with your eyes — it's a miniature composition in clay that stood out among handcrafts exhibited last week at the Middletown Grange Fair.

subset: commoncrawl


text:
Request a Deal
Love this business? The more people who sign up, the more likely we are to get a deal.
People make it happen. Spread the word to request a deal!
13 signed up for this.
About this Business
- HoursSun12:00 PM - 6:00 PMMon-Fri10:00 AM - 7:00 PMSat9:00 AM - 6:00 PM
From Our Editors
A visitor to Diamante Day Spa won?t be hurting for options: between the spa?s 20 different massage styles and 18 distinct facials, clients could be forgiven for having a hard time knowing which treatments would benefit them most. That?s why it?s fortunate that holistic practitioner and spa founder Tara Darlene Diamant? Canhos is not only a skilled massage therapist, but also an experienced lifestyle coach and spiritual consultant. Canhos and her staff utilize this experience to help clients devise the perfect array of treatments. Specialty services encompass mainly Eastern holistic healing practices?chakra balancing, yoga nidra meditation, Tibetan reflexology?which is why it?s fitting that the spa is also home to a zen lounge area complete with fresh fruit, organic herbal teas, and no howler monkeys.


text:
My daughter, Lulu, recently spotted a shirt that she loved – a dip-dyed watermelon number that set her six-year-old heart a flutter. And bless her heart, I gave her the dreaded "I could totally make that" answer. If you were raised by crafty people, you've probably heard that one before. But I held my end of the bargain by creating this simple and oh-so-adorable watermelon shirt that she will wear all summer long. Watch the video below to see how you can make your own just in time for the Fourth of July.
Dyeing fabric may seem daunting, but it's so easy! I think this treatment would be really cute on a picnic blanket as well, don't you? We are just all about some watermelon around here today!


text:
Thus a trip to the Emerald Isle, where folks go around kissing the Blarney Stone, has always been on our traveling wish list. So when our daughter, who lives in London with her husband and two young sons, suggested renting an Irish cottage for a week of transgenerational bonding, my wife and I went for it.
The Davillaun Cottage turned out to be a rather resplendent two-story house in County Mayo, on the northwest Irish coast on the outskirts of the harbor town of Westport.
It included four bedrooms, two bathrooms, a comfortable living room with a fireplace, an enclosed garden terrace, complete kitchen, and washing machine and dryer -- room enough for four adults, plus 4-year-old Benjamin and 1-year-old Gabriel.
We welcome your feedback.
Privacy Policy
Your information will not be shared or sold without your consent. Get all the details.
Terms of Service
JewishJournal.com has rules for its commenting community.Get all the details.
JewishJournal.com reserves the right to use your comment in our weekly print publication.


text:
Visit our other Wisconsin Historical Society websites!
Refine or narrow your results by clicking any term in the gray boxes below.
Expand your results by removing any breadcrumb in a gray outlined box above. Click on the blue "X" to remove the term.
Narrow your results by entering a keyword into the search box above. Then click the "Search Within" checkbox, then click on the orange "Search" button.
If you did not find what you are looking for or you did not get any results, try changing your search term(s). See our Search Help page for more tips.
Date: 1825-08-19
Description: Second page of the Treaty between the United States and the Chippewa, Sauk, Fox, Menominee, Iowa, Sioux, Winnebago and a portion of the Ottawa, Chippewa, a...
Date: 1832-06-23
Description: Document written by Ebenezer Brigham listing events at Fort Blue Mounds.
Description: Third page of the Treaty between the United States and the Chippewa, Sauk, Fox, Menominee, Iowa, Sioux, Winnebago and a portion of the Ottawa, Chippewa, an...
Date: 1972
Description: Letter from Martha MacMillan of Farmington, Michigan to the group Women for a Peaceful Christmas written on stationery bearing an Earth Day logo.
Date: 1821-08-18
Description: Green Bay treaty, August 18, 1821. The treaty is signed by the chiefs of the six Indian nations for the possession of lands near the Fox River to the Winne...
Date: 1974
Description: Angry letter written to Women for a Peaceful Christmas in opposition to their Christmas shopping boycott, signed "A small business owner."
Date: 1952-01-29
Description: The first page of a letter written to A.A. Schecter by Sylvester "Pat" Weaver.
Description: A certificate admitting Lucius Fairchild into the Society of the Army of the Potomac.
Date: 1897-08-28
Description: Receipt for trees and shrubs purchased from Commercial Nurseries of Council Bluffs, Iowa, by Nina Dousman for gardens at Villa Louis.
Date: 1924-12-16
Description: This is page one of a letter written to Governor Emanuel Philipp from Helen Keller. She asks for his assistance in helping establish an endowment fund for...
Date: 1786
Description: Letter written by Gilbert Imlay to Daniel Boone.
Date: 1971 ca.
Description: Page 13 of a booklet published by Women for a Peaceful Christmas offering alternatives to buying Christmas gifts. The page is decorated with several Christ...
Date: 1827-06-22
Description: Front page of "Freedom's Journal", Volume 1, #15.
Date: 1821-07-03
Description: Two pages of a letter written by Thomas Forsyth.
Description: The second page of a letter written by Sylvester "Pat" Weaver to A.A. Schecter.
Description: Letterhead of the Wadhams Oil Company, with an illustration of various red and yellow oil containers against a black circle. The motto, "Emphatically Indep...
Date: 1938-03-09
Description: Confidential letter from President Franklin Roosevelt to United States Minister to Ireland, John Cudahy, on White House stationery regarding events in Engl...
Date: 1860-04-23
Description: Receipt for trees and shrubs purchased from John M. Dunlop by Hercules Dousman for gardens at Villa Louis.
Description: This is page two of a letter written to Governor Emanuel Philipp from Helen Keller. She asks for his assistance in helping establish an endowment fund for...
Description: Letter from Carol Lampe of Fairfield, Connecticut to Women for a Peaceful Christmas written on colorful stationery with a writing surface in the shape of a...
If you didn't find the material you searched for, our Library Reference Staff can help.
Call our reference desk at 608-264-6535 or email us at:
© 1996-2015 Wisconsin Historical Society, 816 State Street, Madison, WI 53706


text:
Great friendly service , very good food. Excellent view of Lake Champlain and Vermont. Also good place for train watching.
Came here last year and were welcome with open arms. Came back this year and received the same reception. Friendly people, great service and good food at a nice price.
We stopped in for lunch and were very pleased with our homemade meatball sandwich. Afterwards, we were told by the locals that his is a good place to eat.
Couldn't ask for more food was excellent and so was the service doesn't look like much from outside but was good real food that tasted awesome
This was an acceptable restaurant, with a view of Fort Ticonderoga. The outside is not very appealing, and you wonder whether you should really eat here. Once inside, you can see the fort very clearly, which is nice. Most of the food is fried, so it's also not really for the healthy crowd. We had the fish basket, and it...
The Fort View Inn is a favorite place to eat. It has a wide selection of food on the menu and we usually have a glass of wine with our meal. The meals have signature twists to old favorites and they are delicious. The owner is almost always there and works along with her employees. It is best to get...
Some of the best wings in the United States. Cathy makes her own secret sauce from mild to extra hot. Her cheesecakes alone are worth the visit. Clams, sandwiches, steaks seafood, wonderful little place
We have eaten here 3 times this week because the food is awesome. We tried other places in this area and this is the best. Do not pass this up. The owner Kathy is so nice and so full of information about this area. do not pass this up
From the moment we walked in and felt the warmth of the Adirondecks we liked it. Katie greeted us with a big smile and seated us immediately. The place is full of locals and very few tourist. The Key West shrimp were excellent as were the bacon wrapped scallops. Katies soup is second to none. All dinners come with a...
Really enjoyed our stop here. Food was very good, even though a bit pricy for the atmosphere.
Bar drinks were good, but also think a bit price for the atmosphere.
Steamed clams were quite good, so were the salads and entrees.
Service was very nice, very friendly.
They are one of the few places open quite late in the area.
Register now for free — and start getting automatic notification of new reviews, responding to traveler feedback, adding new photos to your listing and much more.


text: Over 200 Years of Japanese UFO Sightings A while ago, Hashi wrote a post about Utsuro Bune: Japan's First Modern UFO Encounter. Since then, there have been many UFO sightings in Japan, with most of them having a lot in common and appearing in the same areas. So … [Read more...]Up Next: Scary Needles and Japanese Health Care


text:
I'm waiting for the Eddie Murphy and Jennifer Tilly overlay! ;-)
The Mur
Two different worlds.....we live in two different worlds
I was going to go to the 40th celebration of HM on September 9, but the packages sold out pretty fast.
The movie was okay, but I feel that it made a joke of the Haunted Mansion. I only liked the subtle ride homages.
I was at DL for a bit today and noticed that the Pixie Hollow queue is completely behind walls again. The actual meet and greet is still open, but the queue is down.
Anyone know what's going on? I was surprised given it has just opened.
This is not all that unusual. After being opened for a while, they either noticed some places in the queue that need more work or possibly will redo how the chains are laid out.
While movie tie-ins generally suck, I wish Haunted Mansion would have gotten the Gore Verbinski POTC treatment. I think an era-specific story using the Master Gracey storyline and NO Eddie Murphy would have made for a fantastic movie.
As for no HMH, I don't really think the market of those who want Haunted Mansion to stay "normal" for the 40th is that large. People love that ride, especially considering you can hardly get to Splash Mountain during the holiday months.
"You tried your best and you failed miserably. The lesson is, never try." ~ Homer Simpson
Avatar designed by Greg Maletic
Does anyone know any information on a possible Matterhorn refurbishment, or the rumored "new cars." I know it was closed a few years back for refurbishment and inspections, but does anyone have knowledge of a drastic change?
Based on information from Al Lutz' latest update, I've added an entry for the upcoming Rivers of America refurbishment in early 2010, which will put several attractions out of commission.
MiceAge 4/21: Return to Flash Mountain
Dragon Fly
[ . . . ] the Rivers of America is scheduled to be drained for refurbishment for several months in the winter and spring of 2010, with Fantasmic! returning for the 55th Anniversary summer in June, 2010.
Major Rivers of America refurbishment
Details: Major Rivers of America refurb, resulting in a hiatus for Fantasmic! and a closed Tom Sawyer Island, Sailing Ship Columbia, and Mark Twain Riverboat
Project Status: Drained and closed during winter and spring 2010.
Open Date: June 2010


text:
Nick Van Evelingen (From Falmouth Packet)
- News feed
- Find us on Twitter
All the latest news and views from the Packet
All the latest sport from the Packet
- Find us on Facebook
The Packet
Like us on Facebook
- Search notices
- Birthdays
- Births
- Deaths
- Exams
- Greetings
- Marriages
- Anniversaries
- Other
- Click2Find
Book an announcement for publication online and in the next edition of the Falmouth Packet
40th Birthday
Nick Van Evelingen
Published in the Falmouth Packet on 4 January 12
NICK VAN EVELINGEN - 40 07.01.12 HAPPY BIRTHDAYWith all our love from Mum, Dave and all the Family x x x x
This notice has 197 views, no gifts and no messages.
There are no messages for this notice.
Leave a message
Log in with
There are no gifts for this notice.
Offer a virtual gift
Fields marked with * are mandatory.
Remember you are personally responsible for what you post on this site and must abide by our site terms. Do not post anything that is false, abusive or malicious. If you wish to complain about a particular message, please use the 'report this post' link.


text:
Irish coffee cheesecake
You can't beat cheesecake. It's so rich and creamy and delicious. Some people think it's tricky to make, but it's not. It's basically cream cheese, sugar, and eggs. Nothing complicated about that. I've made Irish creme cheesecake over the years, sometimes with a chocolate swirl, and it's always a big hit. I also love a little Irish creme in my coffee on occasion, so in honor or St. Patrick's Day, I decided to add a layer of coffee flavor to the cheesecake I made for my Irish sweetie. It was the perfect ending to our dinner of reubens and oven fries, and it was even better straight out of the fridge the next morning for breakfast, along with a big cup of coffee. Slainte!
15 graham crackers
4 Tbsp. melted butter or margarine
4 8-oz. pkgs. cream cheese, softened (I used the 1/3 less fat variety and you can't tell the difference)
1 cup sugar
1 tsp. vanilla
4 eggs
2 Tbsp. cocoa
1 Tbsp. strong coffee or espresso
1/3 cup Irish creme liqueur
Crush the graham crackers into crumbs. You can use a food processor to do this quickly and easily, but I find putting the crackers in a large resealable plastic bag and beating them into crumbs with a rolling pin is a good way to release stress. It's also a good job for the kids. Add the melted butter and combine well, then press onto the bottom and up the sides of a springform pan.
In a large bowl (I used my stand mixer), beat together the softened cream cheese, sugar, and vanilla. Add the eggs, one at a time.
Transfer half of the mixture into a separate bowl. Into one half of the mixture, add the coffee and cocoa and stir well to combine. Add the Irish creme liqueur into the other half and stir well.
Pour half of one of the mixtures into the crust, then half of the other, and repeat, making layers in the pan. Run a knife through the batter in a tic-tac-toe pattern to create swirls.
Bake at 350 degrees F. for 40-45 minutes, until the center is set. Let cool completely before removing the sides of the springform pan.
Here's a little hint for making cheesecake: Sometimes you'll get cracks in the top once the cake cools. It doesn't affect the flavor at all. The best way to remedy this is to make a chocolate ganache and pour all over the top, or if you've made a plain cheesecake, open a can of cherry pie filling and pour it over the cheesecake. Nobody will know the difference!


text:
Chicken a L'Orange
|Chicken breast halves
|3 Pound, skinned & boned (6 halves)
|1⁄2 Teaspoon
|Reduced calorie margarine
|1⁄4 Cup (4 tbs)
|Unsweetened orange juice
|1 Cup (16 tbs)
|Dried tarragon
|1 Teaspoon
|Corn starch
|1 Teaspoon
|1 Tablespoon
|Slivered almonds
|2 Tablespoon, toasted
1) Peel the oranges and cut the rind into thin strips. Set aside.
2) Preheat oven to 350° F.
3) Sprinkle the chicken with paprika and saute in margarine for about 4 minutes on both sides or until light brown in color.
4) Take a 13- x 9- x 2-inch baking dish and place the chicken in it.
5) Add tarragon, orange juice and orange rind strips.
6) Cover and bake for about 25 minutes or until nicely done.
7) Remove chicken and set aside. Keep warm.
8) Take a small saucepan and pour the orange juice mixture in it.
9) Take a small bowl and combine water with cornstarch in it. Stir until nicely blended. Add to the orange mixture.
10) Cook until mixture comes to a boil, stirring constantly.
11) Cook for another 1 minute, stirring constantly.
12) Sprinkle the chicken with almonds and serve with sauce.


text:
Hence the animus of former candidate and Texas Governor Rick Perry, candidate Ron Paul, and even Mitt Romney.
But his lack of animus also comes from his post-war experience, one day in particular.
In their meeting, Mao made explicit both his patience on Taiwan and animus against Moscow.
1820, "temper" (usually in a hostile sense), from Latin animus "rational soul, mind, life, mental powers; courage, desire," related to anima "living being, soul, mind, disposition, passion, courage, anger, spirit, feeling," from PIE root *ane- "to blow, to breathe" (cf. Greek anemos "wind," Sanskrit aniti "breathes," Old Irish anal, Welsh anadl "breath," Old Irish animm "soul," Gothic uzanan "to exhale," Old Norse anda "to breathe," Old English eðian "to breathe," Old Church Slavonic vonja "smell, breath," Armenian anjn "soul"). It has no plural. As a term in Jungian psychology for the masculine component of a feminine personality, it dates from 1923.
animus an·i·mus (ān'ə-məs)
An animating or energizing spirit.
Intention to do something; disposition.
A spirit of active hostility; ill will.
In Jungian psychology, the masculine inner personality as present in the unconscious of the female.
["Constraint-Based Animation: The Implementation of Temporal Constraints in the Animus System", R. Duisberg, PhD Thesis U Washington 1986].


text:
Last fall in Barcelona, Spain two PM's from the C# team gave talks on key parts of the new LINQ technology found in Visual Studio 2008:
Luke Hoban was the PM who drove the development of LINQ to Objects, and the man who created the now famous implementation of a ray tracer in a single, very long, LINQ query expression. He is now focusing his efforts on F#. By listening to his talk on C# 3.0 features you will get a chance to see a detailed and specific analysis of how LINQ works. Listening to him give this talk marked a turning point in my understanding of LINQ. For the first time I understood how the team folded extension methods and query expressions together to create the marvelously supple and deceptively simple syntax that we know as LINQ.
Figure 1: Luke Hoban's vision of the inner workings of LINQ to Objects gives developers significant insight into the beauty of C# 3.0.
Luca Bolognese is the Principal Program Manager Lead on the C# team. His talk on LINQ to SQL endows developers with the knowledge necessary to wield the tools that drive data driven application development under C# 3.0. A consummate speaker, Luca skillfully guides his audience through the elements that comprise LINQ to SQL development.
Here are the links you need to get started.
The Talks discussed in this post:
More talks from the TechEd Developers Event Link:


text:
San Francisco ElevatorPitch
- Business & Professional
- 1355 Market Street, #488, San Francisco, CA, 94103
April PitchForce
Tuesday, April 2, 2013 from 5:00 PM to 8:30 PM (PDT)
PitchForce is a part of a series of monthly fund raising events for emerging technology companies in the software, social, mobile, big data, consumer web, finance/payments, clean tech, biotech or medtech sectors. Each month ten companies are chosen to participate in PitchForce. Presenting companies will each have a demo table and have the opportunity to meet and network with angels, VCs and guests during the first hour. Attendees will then vote for their favorite company and the top five will get to present to a distinguished panel of VCs. The panel then chooses who they feel is the most promising startup. The winner will get a meeting with the Sponsoring Venture Capitalist.
Alminder delivers real-time, contextual knowledge to business consumers via a smart mobile calendar app.
Fabbit is a fun, easy, social design platform for the 3D printing market, giving customers the power to create their own products on demand.
FotoNotes innovative mobile and cloud based platform transforms the way that enterprises collect, manage and share mobile information.
Froovie Organics is an innovative organic beverage dispensing solution that combines fruit, vegetable and grains into a "Drink with BenefitsTM"
Ittavi is expense management & bill payment system for child support - ending child support conflict while saving parents time & money.
LoYakk enables Instant "Shared Experience" Communities - on-mobile and on-demand.
Perkville helps small and medium sized businesses retain customers by providing them with a cloud enabled loyalty platform.
Picture Marketing FotoZap® turns consumers into Superfans: Fans that actively contribute social media to any brand's Facebook page or preferred social channel.
PIXT is a fun and easy way to make collaborative walls of photos and videos with your friends and family.
RealtyShares is a real estate crowdfunding marketplace where Accredited Investors can passively invest in pre-vetted real estate deals.
Suraj Kumar Rajwani
Suraj Kumar Rajwani is a co-founder and partner of DoubleRock. He has over seven years of management, investment and mergers and acquisitions experience in Silicon Valley, China and India. He was previously the Managing Director of the Global Network Entrepreneurs Network Organization (GENO), an entrepreneur organization with over 27 chapters worldwide. As head of sales and marketing, he grew GENO's footprint to over 23 countries and led product expansion into new areas such as enterprise pricing and enterprise incentive management. He has advised on mergers and acquisitions in multiple industries including IT, financial services, real estate, and construction. During his tenure at GENO, he developed the GENO Venture Connect framework which served as a platform for entrepreneurs to meet with potential investors which led to the birth and expansion of many successful enterprises.
DoubleRock was founded with the mission to help entrepreneurs achieve their goals of building big, transformative businesses. We are entrepreneurs ourselves and we understand the challenges and skill sets needed to scale good companies into great ones. We formed DoubleRock after several years of doing our own angel investing with a shared belief that the strategy that has succeeded with many of our portfolio companies could effectively be applied towards a wider scope of companies.
Steve Bernardez|Onset
Pete Mayer|LeClairRyan
Suraj Kumar Rajwani|DoubleRock
Paul Weiss|Wylan Capital
Ron Weissman|Band of Angels
Allan Young|Runway
PeopleConnect is a full-service executive search firm specializing in helping innovative startups, early stage ventures, and companies in expansion or turnaround mode grow great teams.
In order to purchase these tickets in installments, you'll need an Eventbrite account. Log in or sign up for a free account to continue.


text:
I have a three year old daughter who is the light of my life! I love her more than life itself. She has always been advanced in everything that she has done.
I just gave birth to my son January 3 ,2010. He was five weeks early which caused us to have to stay in the hospital for twelve days before coming home.
During this time, my daughter started having "Accidents" in her big girl panties. I figured once mommy came back home all would get back to normal.
She loves her new baby brother and shows no signs of jealousy. She gets just as much attention now as always, however she is still having accidents in her panties as well as getting very mouthy and talking back alot. She does not listen when told to do something or takes an hour to do it. Her arguing is getting out of control and I personally don't know what to do.
We have popped her rear end, put her in time out, taken away movies, toys, and anything we can find. Nothing seems to work. I tried giving her a goal to work towards. Showed her a calendar, marked a date two weeks from the other day and said if you can be a big girl and get smily faces for the all of these days, then you get to go to this show on this date. She got really excited and the next day she got a sad face for having an accident in her panties and not listening! I thought for sure giving her a positive goal to work for would work better than all the negatives of punishment you know. But I guess NONE of these options seem to be working!
I just want my little girl back, I have never had these issues with her before and between her acting out and having a newborn in the house again, I am just becoming over whelmed and running out of options to help.
Any one with any kind of ideas, please post them! I am will to try just about ANYTHING at this point to get my daughter back on track.


text:
Patrick Fischer, a retired computer science professor at Vanderbilt University whose secretary was injured when he was targeted by the Unabomber in 1982, has died. He was 75.
Fischer died Aug. 26 in hospice care in Montgomery County, Md., the university announced. A cause was not given.
And he was an early target of the Unabomber, the name that had been given to a perpetrator who had killed three people and wounded at least 23 from 1978 to 1995. The bomber was later identified as Ted Kaczynski.
On May 5, 1982, Fischer's secretary, Janet Smith, opened a package addressed to the professor and a pipe bomb stuffed with smokeless powder and match heads exploded in her hands. She was hospitalized for three weeks with burns and cuts to her chest, arms and hands.
Fischer, who was attending a conference in Puerto Rico at the time, said he could not recall meeting Kaczynski.
Authorities eventually linked the incident to Kaczynski, who is now serving a life sentence. He was not charged in the Vanderbilt incident because the statute of limitations for such a crime had expired by the time he was arrested in 1996.
For years afterward, Fischer wondered how the mail bomber had chosen him.
"For a while I thought he opened up 'Who's Who' and threw a dart until he got a computer scientist," Fischer told The Times in 1996. "That's probably wrong. There has to be a link."
Fischer did notice odd coincidences. His father was a math professor at the University of Michigan when Kaczynski was a graduate student there. And Fischer was a math student at the Massachusetts Institute of Technology while Kaczynski was studying across town at Harvard University.
"The thing is, yes, we knew a lot of the same people," Fischer told the Dallas Morning News in 1996. "But I don't think I know him directly. I didn't recognize the face, either the shaven face or the bearded face. I don't recognize the name at all."
Patrick Carl Fischer was born Dec. 3, 1935, in St. Louis. He received a bachelor's degree from the University of Michigan in 1957 and a doctorate in math from MIT in 1962.
He taught at Harvard, Cornell University, the University of Waterloo in Canada and Penn State before moving to Vanderbilt.
Kaczynski originally addressed the package to Fischer to Penn State, and it was forwarded to him at his new position in Nashville.
Fischer never again assumed a package would be safe to open, telling the Times in 1996: "I will be careful for the rest of my life."
Survivors include his second wife, Charlotte Froese Fischer, a professor of computer science at Vanderbilt; a son and a daughter.
Patrick Fischer dies at 75; target of Unabomber
A computer scientist at Vanderbilt, Patrick Fischer was out of town when his secretary opened the package bomb, setting off a blast that injured her.


text:
I am terrible at Jquery, and can barely understand it as I try to make this code for my site.
My goal is to have Div tags appear when the value that is selected in a form equals that Div tag's name.
Here is what I have:
<select name="SELECT_Mood_30">
<option value="hap">Happy</option>
<option value="sad">Sadness</option>
<option value="ang">Angry</option>
<option value="gru">Grumpy</option>
<select name="SELECT_hair_75">
<option value="red">red</option>
<option value="bla">black</option>
<option value="blo">blonde</option>
<option value="bru">brunette</option>
Now imagine there's maybe 40, 50, or even 100 different options. Each with a hidden div like below. Upon selecting the option with a value that matches the name of one of these divs, I want the div to appear. When deselecting it I want it to disappear.

How can I make you feel better?

Black hair is really cool

Red hair is nice!
Is it possible in Jquery to make this? Possibly make it look up the value that is selected, then unhide the named div. The name of the form doesn't matter, it could look up the color red in the mood form and if it appears it could display it anyway. If the named div doesn't exist, then nothing happens and all is good? (no error message or anything)
I've found a few working ones online, but they all seem to require to put each option inside the Jquery, which is what I don't want. I would like it to be flexible enough where you simply have to change the option values and the div names.
Edit: I forgot to mention that say if one select is chosen, such as "sad", and then someone chooses "red", I don't want the div that was named "sad" to disappear. but if the "ang" option is selected, I want the "sad" to disappear. I hope that doesn't confuse things a lot more.


text:
You may also like
Rate Dr. Colin Birch
Dr. Colin Birch's Ratings
I really appreciate Dr Birch. He is a genuine man who is a very smart and Compassionate OBGYN. Dr B takes good care of you and makes you feel completely relaxed at your visits or when it is surgery time. Thanks for all your devotion to me and your professional work ethics.
Was this rating useful? 0 [[ userState['ratings'][2683135].votes_count ]]
Dr. Birch and his staff were amazing. Made me comfortable right from the beginning which was difficult as I am an under 40 year old woman with ongoing bladder issues due to having children. I would recommend him to anyone. Fantastic. Delightful staff. I barely had time to open a magazine and they were calling my name to come back. Within minutes of changing the Doctor and Assistant were in speaking with me.
Was this rating useful? 0 [[ userState['ratings'][2639496].votes_count ]]
I had DR birch for a biopsy from an abnormal pap. I was very nervous and feeling quite uncomfortable and exposed. It was nice he explained what he was going to be doing exactly. But when he commented that he was applying the iodine solution to my cervix so if anyone commented that I smelt like "fish and chips" afterwords I would know why... Well I felt even more uncomfortable. Perhaps this joke was supposed to lighten the mood but I thought it was very unprofessional and rude.
After the biopsy I was left to redress.
When I stood up from the bench a gush of blood came from inside and landed on the floor. I didn't know if this was a normal thing to have happen. I was given a larger than life jumbo sized maxi pad to use following the biopsy but nothing was mentioned about this much blood. There was a bathroom in the DRs room I had the procedure so I went and sat on the tolit and I continued to bleed more. After about 10mins dr birchs assistant knocked on the door to see if I was alright. She helped to calm me down and told me that I had nothing to worry about.
I wish that I could have been more prepared for this.
Was this rating useful? 0 [[ userState['ratings'][2600162].votes_count ]]
Wow...the best surgery ever..this Dr is amazing. Painless surgery. Fixed Blatter. He is a god sent ladies.. We need more like Dr birch
Thanks again
Was this rating useful? 0 [[ userState['ratings'][2599811].votes_count ]]
I was lucky to have Dr. Birch as my doctor for my second pregnancy. After complications with my first, I really appreciated his honesty, knowledge and clearly presented options including risks/potential issues and benefits. He was always frank, professional and approachable.
Was this rating useful? 0 [[ userState['ratings'][2583795].votes_count ]]
I'm 24 and a Hypochondriac. When I discovered that I had to get a biopsy done due to an abnormal pap, I was terrified. Dr Birch's RN Kim and I got in touch soon after. She was very polite and reassuring. She also did her best to make me smile because she could tell I was terrified. Meeting Dr Birch was intimidating at first, but then he explained to me what he would be doing and made me feel comfortable. He told me immediately that i didn't have cancer which helped with my anxiety. I've met with him two more times sonar and he is excellent. He was comfortable asking any OB/GYN related questions I had, even ones that had nothing to do with me. I discovered through my pestering that he has worked i third world countries and has seen some horrible things. I left with a new respect for him. He is very sweet, and even though he is quiet, it doesn't mean he is arrogant or rude. I highly recommend Dr Birch and his sweet RN Kim
Was this rating useful? 0 [[ userState['ratings'][1253892].votes_count ]]
I'm surprised at all the positive comments on this doctor. I wonder if his attitude is different based upon the reason for seeing him and whether it's in his office or in hospital. He's been the OB on call twice now since I've been on hospital bed rest and has not once actually told me who he was. I had to ask my nurse who he was since she was in the room the 2nd time he came. This man is, I'm sure, a perfectly competent doctor but his attitude needs some adjustment. He walks in the room as though he is god's gift to pregnant women and we should all fall at his feet and grovel. His first visit to me consisted of a whopping 2 minutes where he tried to push getting a shot of blood thinners every day on me even though the MFM team advised I didn't need it, including the head of the department and head of U of C's Obstetrics department since my bed rest does allow for walking around a few times each day.
Was this rating useful? 1 [[ userState['ratings'][1253891].votes_count ]]
I first met Birch when I had an emercancy c-section with my daughter in 2010. He was very helpful and calmed my nerves about a c-section and talked to my husband about his concerns. When I found out a year and half later I was pregnant agian I requested Dr Birch without hesitation. He is the best doctor I have ever met. I moved to BC half way through my pregnany and I am driving to Calgary every 2 weeks I believe in him so much. He tells you the facts as it is. He will not hold your hand through every step but he will make sure you are comfortable and understand. Whenever I go see him his staff is great and polite and he is sitting there waiting for me. He gets my daughter involved in the appointments and always makes her laugh. He is a great doctor and I am very comfortable with him doing the surgery and I am proud to say both my kids will be born by Dr. Birch, this doctor is amazing.
Was this rating useful? 0 [[ userState['ratings'][1253890].votes_count ]]
I recently had a partial hysterectomy with Dr. Birch in the Canmore hospital (shorter wait times). I cannot say enough about his bedside manner and skill. I felt very comfortable in his care. He told me that is I had any questions prior to the surgery date that I was to come back for another visit. He felt that if I was going into surgery with any unanswered questions that he was not doing his job! How refreshing to have that said to prior to a surgery. Not too sure why he gets any negative feedback as he has always been pleasant to deal with. My only complaint is of the staff at the Pelvic floor clinic admin staff are not very attentive, nursing staff has been great. Sue at the clinic is awesome to deal with!!!
Was this rating useful? 0 [[ userState['ratings'][1253889].votes_count ]]
I had an emergency csection with my first child and he was very unprofessional. He used lots of stupid words like cool, awesome, etc. I felt like I had some high school student performing my surgery. In the end I ended up with a terrible infection that I had to be re-hospitalized for, separating me from my newborn for 3 nights, it was terrible. He didn't even book me for a follow up. When ALL other doctors do that, especially after major surgery. I ended up seeing Dr. Cenaiko, who is awesome, for my follow up, he was also the doctor that I saw during my extra 4 days in the hospital while I had my infection. But back to Dr Birch, NEVER would I have him deliver another child of mine, I felt that he didn't care or have any real knowledge.
Was this rating useful? 0 [[ userState['ratings'][1253893].votes_count ]]
Top Hospitals in Calgary, AB
Find a Doctor by Specialty
You may also like
Top Gynecologists (OBGYN)
#1 of 145
#2 of 145
#3 of 145
#4 of 145
#5 of 145


text:
Former NFL quarterback Ryan Leaf has been transferred to Montana State Prison after being kicked out of a drug treatment center for allegedly threatening a staff member.District Judge Kenneth Neill ordered the former San Diego Charger to serve a five-year commitment to the Montana Department of Corrections, state officials confirmed on Thursday. Leaf had been in "lockdown" at the drug treatment center since July 12. The 36-year old was terminated from the treatment program and subsequently placed behind bars after he was found guilty of "behavior that violated conditions of his drug treatment placement." One of the violations included threatening a staff member, said a regional probation and parole administrator.According to the DOC's Correctional Offender Network Search, Leaf's status last changed Jan. 10, and it lists his current location as the reception unit at the Montana State Prison.The program is intended to offer rehabilitation assistance for those addicted to methamphetamine or any other addiction with a co-occurring disorder.Leaf was drafted No. 2 overall by the Chargers in the 1998 NFL Draft.Via Great Falls Tribune
NFL News
Delivered to your inbox
You'll also receive Yardbarker's daily Top 10, featuring the best sports stories from around the web. Customize your newsletter to get articles on your favorite sports and teams. And the best part? It's free!
|Best of Yardbarker
|NFL News
|MLB News
Today's Best Stuff
For Bloggers
Join the Yardbarker Network for more promotion, traffic, and money.
Company Info
What is Yardbarker?
Yardbarker is the largest network of sports blogs and pro athlete blogs on the web. This site is the hub of the Yardbarker Network, where our editors and algorithms curate the best sports content from our network and beyond.
|© 2015 Yardbarker, Inc. All rights reserved. Use of this website (including any and all parts and components) constitutes your acceptance of these Terms of Use and Privacy Policy. (What's New?)


text:
'Voices & Thoughts' Archives
The Bible tells me so
Dalai Lama, Jason Collins, Leviticus and You. By Robert De Filippis From the Dalai lama, "The purpose of religion is to control yourself, not to criticize others. How much am I doing about my anger, attachment, hatred, pride and jealousy? These are the things which we must check in our daily lives." Let's see, how can we fracture that advice this week? Oh, Oh, I know, let's criticize and condemn Jason Collins for being the first openly gay NBA Player. Hey, while we're at ... Full Story
United States, the last empire?
And Then There Was One Imperial Gigantism and the Decline of Planet Earth By Tom Engelhardt It stretched from the Caspian to the Baltic Sea, from the middle of Europe to the Kurile Islands in the Pacific, from Siberia to Central Asia. Its nuclear arsenal held 45,000 warheads, and its military had five million troops under arms. There had been nothing like it in Eurasia since the Mongols conquered China, took parts of Central Asia and the Iranian plateau, and rode ... Full Story
Children and guns, deadly combination
Florida 13-Year-Old Finds Family Gun, Shoots 6-Year-Old Sister Another tragic shooting involving kids. May 6, 2013 | A 6-year-old Florida girl is in critical condition after being shot by her 13-year-old brother, Florida officials say. According to the Florida Sun-Sentinal, the teen found the gun when the siblings were home alone. Although the incident is still under investigation, the shooting appears to have been accidental, the Sheriff's ... Full Story
Obituary: Kenneth L. Fox
Kenneth L. Fox June 25, 1926 – April 23, 2013 Kenneth Lee Fox was born in Zenith, Kansas as the last child in a family of 7 children to Gertrude and Fred Fox. He loved to tell that he was born on the birthday of his oldest brother, June 25, 1926. Ken was well respected and very active in his small farm community as an Eagle Scout, and as an athlete in basketball, football and track. When Ken was in 9th grade his parents had to move to find work so he continued to ... Full Story
America, the land of the destitute
By Paul Buchheit Five Ugly Extremes of Inequality in America -- The Contrasts Will Drop Your Chin to the Floor Any of the ten richest Americans could pay a year's rent for all of America's homeless with their 2012 income. March 24, 2013 | The first step is to learn the facts, and then to get angry and to ask ourselves, as progressives and caring human beings, what we can do about the relentless transfer of wealth to a small group of well-positioned ... Full Story
What happened to Conservatives?
American Right-Wingers Are No Longer Conservative — They're Extremists Their reactionary bent is manifesting itself in legislatures across the country. April 17, 2013 By Joshua Holland College students taking Poli-Sci 101 learn that conservatism is an ideology that reveres established tradition, emphasizes a deep respect for the rule of law and comes with a deep distrust of rapid social change – especially change driven by public policy. William F. ... Full Story
Welcome to Ayn Rand Planet
Why the Rich Prosper from the Stock Market While We Get Screwed By Les Leopold On Planet Rand, the stock-market boom is a wonderful thing precisely because it rests upon the recent rise in corporate profits. March 11, 2013 | This morning I received an email blast form Amazon. It was pushing The Fountainhead and Atlas Shrugged, the old and new testament of the Ayn Rand nation, which includes much of the Tea Party and its Republican allies. How ... Full Story
Obituary: William H. Schwindt
William "Bill" H. Schwindt, 90, passed away on April 16, 2013, at home. He was born on March 6, 1923 in Berthoud to Henry and Elizabeth (Nolde) Schwindt and grew up in the Berthoud/Loveland area. His parents and grandparents were German-Russian immigrants who settled and farmed in Berthoud. Growing up on a farm, the values of hard work and family traditions were taught to him early, and carried through to the rest of his life. He married Dorothy Frank on January 26, 1947 in Loveland ... Full Story
Obituary: Kevin Michael Campbell
Kevin Michael Campbell, 23, of Fort Collins, passed away Thursday, March 28, 2013 in Loveland. Kevin was born August 24, 1989 in Loveland to parents Kent and Candy Campbell. He graduated from Thompson Valley High School and was a senior Biology major at Colorado State University. He was devoted to his studies and deeply enjoyed learning. He aspired to be a professional athletic trainer. Kevin loved Father God with a passion and intensity that is indescribable. His polite, ... Full Story
Blacks uninsured, unhealthy and unemployed
Courtesy of: Financesonline.com - Hot Financial News


text:
LeBron James Scores 61 on Bobcats
LeBron James scored a career-high 61 points, breaking Glen Rice's franchise record, in the Heat's 124-107 win over the Bobcats on Monday night. LeBron James spoke afterwards.
Indiana Pacers Owe Thanks To Roy Hibbert
The Indiana Pacers and their fan base owe thanks to big man Roy Hibbert for everything that he has done for the team and city now that he is being traded to the Los Angeles Lakers. Read More
Lakers Need Bryant To Take Pay Cut
Kobe Bryant is 36 years old, and he is due to make $25 million during the 2015-2016 season. Find out why it's time for him to take a pay cut. Read More
Kings Are Up To Something By Signing Kosta Koufos
The Sacramento Kings made a confusing move by signing Kosta Koufos. Read More
Roy Hibbert Will Suit The Lakers
The Los Angeles Lakers will end up being very happy that they acquired Roy Hibbert from the Indiana Pacers, and he will turn his career around during the 2015-16 season. Read More
Aldridge Will Completely Bolster Spurs
Find out why new acquisition LaMarcus Aldridge is about to make serious waves with the San Antonio Spurs next season. Read More
Raptors Go for Defense Again With Bismack Biyombo
The Toronto Raptors are doing everything in their power to fix last year's leaky defense. Read More
Hibbert Would Fit Nicely With Lakers
After losing out on the LaMarcus Aldridge sweepstakes, the Los Angeles Lakers are rumored to have interest in Roy Hibbert. Find out why he would be a solid addition. Read More
Clippers' Dawson Dazzles In Summer League Debut
Find out why the Los Angeles Clippers should be so happy to have Branden Dawson. Read More


text:
Premium Content From ASCII Members
What Every Financial Services IT Solutions Provider Needs To Know About Cybersecurity
Most IT organizations (both in house and outsourced) that support financial services businesses tend to look to regulations as their rulebook to ensure security and keep the auditors off their backs. However, I take a different approach. Not only should the IT organization support the institution's fiduciary responsibilities but also assist with keeping the company's name out of negative press due to a breach or data leakage.
5 Warnings About Jurisdiction And Cloud Storage
Technology and law are two things that can never seem to get on the same page. Technology moves at such a rapid pace that the law can never seem to catch up. Now, mix in the advent of cloud storage and the complexities begin to mount. Cloud storage has spurned a global phenomenon where your clients data can be housed anywhere in the world. Without even knowing it, your clients data stored in the cloud can be subject to local, national and even international laws.
3 Reasons That Justify Raising Your Rates
This is another in the arsenal of tough lessons that most small business owners learn — charging your customers more. It isn't unusual to be very nervous about raising your rates for fear you will lose a client. In an ideal world, best practices would dictate that you raise your rates every year.
7 Things Your Antimalware Offering Must Do
You would be hard pressed to find an IT professional who does not offer an antivirus solution to their clients; but it is just as crucial to have a solid antimalware offering for your users.
3 Ways To Build Wealth With The As-A-Service Business Model
On my way to work I pass a billboard advertising the Connecticut Lottery. They now have two games available with a "win for life" top prize — very tempting until you look at the odds of winning. Every day I play a different type of game — my "win for life"' strategy is based on recurring revenue streams in my managed IT services business.
Adding These 3 Services Will Build Stickier Customer Relationships
One of the toughest things that any business owner will run into is, "How do I give more value and keep my customers coming back for more? What can I do to keep the relationship going?" If you are a large, big box company, you have worked the process to an art form. To a small or medium business owner that sells widgets, the challenge can be tough. How do you add value to the transaction (bigger sale) or add value to the relationship (more purchases more frequently)?
7 Reasons NOT To Offer Service Package Options
When I started Triada Networks in 2008, I had to decide which billing model made the most sense for me. Since I knew that I didn't want to trade dollars for hours, fixed-price plans seemed to make the most sense, which ultimately led me to embrace the concept of managed services. By Raffi Jamgotchian President/CTO of Triada Networks
How To Fire A Client
As a business owner, one of the hardest things I have had to learn is when it is time to fire a client. I know that I am not the only business owner to face this challenge. The need to find and retain clients is driven by the crucible of those early days when you did not have very many clients — when the business was struggling to survive. One cannot abide to lose a client — even if they are a lousy client. You fret and worry, maybe if you do a better job next time around they will pay their bills on time or be less argumentative?
5 Ways Cloud Services Are Forcing Your Business To Change
Clouds! Public! Private! Hybrid! Buzzwords ... and more buzzwords! Let's face it, cloud computing has arrived, and is not going anywhere. Sure, we had the same technologies "back in the day" (think: mainframes, where access was remote and not on-premise, and secure) and today, everyone professes to have a "cloud" solution.
How To Bundle And Price Managed Services
Sometimes a client will ask me, "What computer should I buy?" I can only answer that question with a series of other questions about their unique computing needs and preferences. If I ask another IT service firm how they bundle and price their managed services plans, I hear entirely different options and pricing from my own plans. Each client is unique in terms of their buying patterns, just as each business owner is unique in their pricing structure.


text:
Begin New Search
Refine Last Search
Cemetery Lookup
Add Burial Records
Help with Find A Grave
Top Contributors
Success Stories
Discussion Forums
Find A Grave Store
Log In
Mary Joan
member for 5 years, 9 months, 24 days
Add to MyFriends
Bio and Links
I joined Find-A-Grave to "pay forward" the help I have received from others.
Most of the cemeteries, in Defiance and Williams County Ohio, have been read/walked and indexed. I would be willing to look up and send information if requested. I am most knowledgeable about the cemeteries in Farmer Twp. and Washington Twp., Defiance County; and for St. Joseph Twp., Springfield Twp. and Pulaski Twp., in Williams County.
OF INTEREST to others may be the link to the Williams County Public Library (WCPL) obituary index. Anyone who has died in the Williams County, Ohio area since 1870 is most likely on the index. Many Defiance County, Ohio obituaries were included in the Williams County newspapers. I suggest searching by using the last name only. Keep in mind the spellings may vary so try several options if you do not get results on the first search. If a name is listed more than once, it is because an obit was published in more than one local paper. Not all of the papers have been added, so check back often to see if there is a new entry for one of your Ancestors.
The obituary index direct link is
http://obits.mywcpl.org/ (updated 01-2015)
You can request obits online and they will be sent by email.
The link to the WCPL library history and genealogy department is
http://www.mywcpl.org/local (updated 01-2015)
The above URL will have links to Williams and Defiance County Genealogy Society websites.
Leave Public Message
Contributions to
Find A Grave
406 Memorials Added
509 Memorials Managed
1 Memorial/week
735 Photos
5 Photo Requests
92 Volunteer Photos Taken
1,019 Virtual Flowers
3 Virtual Cemeteries
2 Friends
Search Contributor's Records
First Name
Last Name
Virtual Cemeteries
His Direct Line Ancestor...
My Direct Line Ancestors
Touching or Unusual Tombs...
Find A Grave Friends
Keith Keller
Lucille Francis
Messages left for Mary Joan (109)
Leave Message
Mary (Lee) McCulley
RE: 88468314 and 88468509
Yes Ella M. Tharp would be a Great Grandma to my husband.
Added by
Mary (Lee) McCulley
on May 22, 2015 6:03 AM
Joanne Thompson Berger
William Mann #66201397
Hello ...I am wondering if it would be possible to use your cemetery photos on my tree . He is my 4x gr grand and I will give you and Find A Grave full credit... Thank you Joanne
Added by
Joanne Thompson Berger
on Apr 19, 2015 6:07 PM
RE: Clara M Conkey Smith
Thank you for creating a memorial for Clara. The only information I have on her is that her son married Carrie Ertel Husted. So I didn't want to create one just to have the link. I have linked her to her son, Ralph.
Also, I will be sending you an email. My email is email@example.com
Added by
on Apr 15, 2015 9:27 PM
George OHLEMACHER
The Woodlawn Cemetery records state this individual is buried in the City Lot. This land is considered a Potters Field and there is a stone marker indicating this. The land is directly south of the office and adjacent to Webster Ave. Less than a dozen markers are visible and most cannot be read, even with a rubbing. Was not able to locate a marker for any of the eighteen (18) requests. Sorry!
Added by
on Mar 25, 2015 6:53 PM
Matilda Tharp
Please link gggrandmother Matilda Tharp to my ggrandmother Find A Grave Memorial# 52178497,
Thank You
Doug Kigar
Added by
on Mar 02, 2015 7:18 PM
Thomas L. Harman
Wayne Hulbert & Beulah McCollough
Thank you so much for posting the engagement photo for Wayne and Beulah.
Always like to be able to put a face to the names.
Added by
Thomas L. Harman
on Feb 04, 2015 1:14 PM
J Mann
William Mann - Kemp Cemetery
Mary Joan - Please contact me at firstname.lastname@example.com.
John Mann
GGG Son of William Mann
Added by
J Mann
on Jan 21, 2015 4:56 AM
Neil B (John 3:16)
Hi Mary Joan,
♪ღ♪*�.���.�*�(�`'�.�(♥)�.� '��)�*�.���.�*�♪ღ♪�**�.
♪ღ♪*�.���.�*��*�.���.�*�♪ღ ♪�.�*��*�.���.�*�♪ღ♪�
Thanks for your friendship. May this year bring you many unexpected blessings.
Added by
Neil B (John 3:16)
on Jan 02, 2015 1:43 AM
Hallie Smeltz Martinson
RE: More Smeltz info to email
2 emails came through on FindAGrave but, no attachments.
You can use my email to send firstname.lastname@example.com
I am dying to see what you have found.
Added by
Hallie Smeltz Martinson
on Dec 29, 2014 1:08 PM
Dotti Lydecker
James Pollock
I didn't get your message and do appreciate why, getting used to a new computer can be frustrating at times. Great news you've found someone that might be able to help, please let me know what I need to do. Thanks so much for your help.
Added by
Dotti Lydecker
on Dec 29, 2014 7:22 AM
View all messages...
Privacy Statement
Terms of Service


text:
Orders must be placed in the US, they cannot be placed internationally.
Browse and select the photo you want to have printed on your gift card.
We review each image submitted. While we accept most images, we will not accept images that contain any of the following:
Big Daddy Leather (BDL) was our second stop while in F-Burg. After visiting Renegade and then eating some lunch, we decided to check out BDL.
Small shop,...


text:
Zebra Wool Rug, Neutral 2 2
great rug! I love animal prints, but some are more tasteful than others. This is one of those - has just enough drama, while not to "showy". The pile is luxurious. It looks great & feels great in front of our gas wall fireplace. March 15, 2015
contemporary quality This is a beautiful accent rug and adds interest to my living space. July 6, 2013
1-2 of 2


text:
Apple processor price hike denied by Samsung
It would appear that the price hike spoken about earlier this week between Samsung and Apple has been denied by a Samsung official - albeit an anonymous one. This official spoke with the Hankyoreh newspaper in Korea, saying that prices were generally agreed upon between the two companies at the beginning of each year for hardware purchased by one and sold by the other, making a change such as the one mentioned all but impossible. This same source noted that such prices "aren't changed easily" and that the 20 percent price jump Samsung was said to have demanded from Apple was likely entirely fabricated.