text:
An enzymatically active reusable sponge or foam, capable of regeneration with oximes, made of a polymer such as polyurethane is prepared for detoxification of hazardous compounds such as organophosphorus and organosulfur compounds. The foam or sponge contains a plurality of enzymes including enzymes selected from acetylcholinesterase, butyrylcholinesterase, triesterase, pseudocholinesterase, organophosphate hydrase, phosphotriesterase, paraoxonase and organophosphorus and organosulfur hydrolyzing enzymes. The sponge or foam may additionally contain activated carbon and an enzyme reactivation compound. A kit can be formed containing the sponge or foam and the compound for enzyme reactivation. The enzymatically active foam or sponge may be prepared using a two chamber device where enzymes and prepolymer are passed from separate chambers into a static mixing stator and are subjected to low shear mixing and extrusion to form by the sponge or foam.
1. A method of making an enzymatically active reusable polymeric sponge or foam, capable of regeneration with oximes, for the detoxification of a hazardous compound comprising immobilizing a plurality of enzymes on or within the sponge or foam by mixing the plurality of enzymes with a polyurethane prepolymer in an apparatus having a static mixing stator connected to both first chamber and a second chamber, wherein the first chamber contains a mixture of the plurality of enzymes and the second chamber contains the polyurethane prepolymer, and equal parts of the mixture of the plurality of enzymes and the polyurethane prepolymer are passed from the first and second chambers into the static mixing stator where the enzymes and prepolymer are mixed under low shear conditions while being rapidly and evenly extruded through the static mixing stator to form said sponge or foam, said plurality of enzymes capable of detoxifying organophosphorous and/or organosulfur compounds, wherein said plurality of enzymes comprises at least one enzyme selected from the group consisting of: acetylcholinesterase (AchE), butylcholinesterase (BchE), triesterase, pseudocholinesterase, organophosphate hydrase (OPH), phosphotriesterase, paraoxonase and organophosphorus and organosulfur (OP) hydrolyzing enzymes.
2. The method of
3. The method of
4. The enzymatically active reusable polymeric sponge or foam for the detoxification of a hazardous compound made by the method of
5. A method of reactivating said polymeric sponge or foam of
6. A method for treating a contaminated surface comprising contacting the surface with the reusable sponge or foam of
7. The method of
8. The method of
9. The method of
10. The method of
11. A kit for the detoxification of a hazardous chemical comprising a reusable a polymeric sponge or foam of
This application is based on Provisional Application No. 60/130,987, filed Apr. 26, 1999.
This invention relates to materials, compositions, kits and methods for neutralizing, detoxifying or decontaminating equipment and/or personnel exposed to organophosphorous and organosulfur compounds.
Methods for decontamination, neutralization and removal of chemicals, such as organophosphorous and organosulfur (OP refers to both) compounds, herbicides and insecticides, are known in the art. However, the compositions and devices utilized in the prior art methods have undesirable properties, such as corrosiveness, flammability, toxicity, difficulty in making and storing, and limited shelf-life.
For example, DS2, a standard decontamination agent, comprises 70% diethylenetriamine, 28% ethylene glycol monomethyl ether, and 2% NaOH by weight. Although DS2 is effective, it is corrosive upon exposure to air. DS2 and any matter resulting from its use is classified and regulated as hazardous material. After an application, the DS2 must stand for 30 minutes before rinsing the treated area with water. Additionally, DS2 comprises a teratogen.
Some decontamination methods employ hypochlorite formulations which are corrosive and toxic and injure humans and sensitive tissues such as eyes. Other methods comprise incinerating the contaminated material and utilizing carbon filters to absorb the residual chemicals. Yet other methods utilize polymer beads or microemulsions which absorb the chemical and must be rinsed away. These methods are inherently dangerous, expensive and generate hazardous waste. Furthermore, as many of these compositions and compounds utilized degrade upon exposure to water and carbon dioxide, these compositions and compounds must be used the same day they are made.
Some in vivo methods employ cholinesterases in the presence of nucleophilic oximes to detoxify OP compounds. This enzyme bioscavenger approach is effective against a variety of OP compounds in rodents and nonhuman primates. For examples pretreatment of rhesus monkeys with fetal bovine serum acetylcholinesterase (FBS-AChE) or horse serum butyrylcholinesterase (Eq-BChE) confers protection against up to 5 LD50 of soman, a highly toxic OP nerve agent. Although, the use of an enzyme as a single pretreatment drug for OP toxicity is sufficient to provide complete protection to an individual subject, a relatively large (stoichiometric) amount of the enzyme is required to neutralize the OP compound in vivo. Therefore, OP/enzyme stoichiometry is increased by combining enzyme pretreatment with oxime reactivation so that the catalytic activity of OP inhibited FBS-AChE is rapidly and continuously restored, and the OP compound is detoxified.
Clearly, a need for better methods and devices for neutralizing, detoxifying, decontaminating and cleaning materials, equipment and personnel exposed to OP compounds exists.
Recently, OP detoxifyng compounds, devices and methods thereof, which allow the safe, effective and convenient detoxification of highly toxic compounds not possible by the prior art, have been developed. These environmentally friendly compounds, devices and methods are disclosed hereinbelow.
The present invention provides materials, compositions, kits and methods for neutralizing, detoxifying or decontaminating equipment and/or personnel exposed to OP compounds.
In one embodiment, the invention relates to a material comprising a mixture of enzymes and substrates for the removal, decontamination and neutralization of OP compounds including those directed against humans. The mixture of enzymes utilized comprises cholinesterases (ChEs) and/or OP hydrolases and reactivators, such as oximes which includes mono-disquarternary oximes. The material may comprise a flexible or rigid porous support. The porous support may be a polyurethane matrix or equivalent.
For example, the porous support may be a flexible sponge-like substance or like material, wherein the enzymes are secured by immobilization. Depending on the polyurethane prepolymer or substrate utilized, porous supports of varying degrees of flexibility and porosity may be obtained. The porous support may be formed into various shapes, sizes and densities, depending on need and the shape of the mold. For example, the porous support may be formed into a typical household sponge or a towelette. The preferred dimensions of the sponge are 1″�2″�8″ to 2″�4″�8″. The preferred dimensions of the towelette are 4″�4″�0.25″ to 4″�4″�0.03125″ to 14″�14″�0.0625″. However, during large-scale synthesis, the dimensions of the initial immobilized enzyme product might be large. For example, approximately 4 feet by 8 feet rolls could be produced and sized as appropriate and described above. The sponge-like support would be preferable for use on surfaces, including natural, synthetic and biological surfaces such as equipment, laboratory hardware, devices, skin and other delicate membranes, where decontamination of a rough or irregular surface is desired or where the prior art decontamination materials are incompatible with human tissue. For example, the materials may be used to clean and decontaminate wounds as it is non-toxic and the immobilized enzymes will not leach into a wound. Therefore, the sponges could be used to decontaminate civilians contaminated by a terrorist attack at a public event.
If an object and/or area to be neutralized or decontaminated comprises cracks, crevices, porous or uneven surfaces, a foam-like support is suitable. Application of small quantities may be done with a spray-bottle or spray can with an appropriate nozzle. Further, foam may be selected so that it can be dispensed into the opening of sensitive equipment or an orifice of a subject, such as the ear canal. If a large area is contaminated, an apparatus that dispenses a large quantity of foam may be utilized.
The foam-like support may dissipate after a period of time like shaving cream or it may cure into a stable and flexible sponge-like support. The dissipating foam may be applied on living subjects. The foam, which cures, may be applied around an object and contain the contamination within the foam. Once the foam cures, the object may be handled and moved without further exposure to the hazardous chemical.
When necessary, the material may also comprise a rigid and porous support. The rigid material can be ground into a powder and added to lotions, soaps and other liquids for application. Likewise, the flexible material, supra, may be appropriately treated to render it suitable for use in lotions, soaps and other liquids.
The material may also be in the form of a filter for neutralizing, detoxifying or decontaminating gases such as air. Additionally, the material may be in a form suitable for use as clothing or linings of clothing. Furthermore, the material may be used to decontaminate water by placing the material in water and then removing it from the water.
In another embodiment, the material can be color-coded according to the specific substance it may neutralize, detoxify or decontaminate. The color or color scheme could be selected to indicate enzymatic concentration, activity and/or remaining shelf-life or range thereof.
As disclosed herein, one of ordinary skill in the art will appreciate the various materials and their uses as contemplated by the inventors. All of these forms may be appropriately combined with carbon for further absorption of OP compounds. The carbon may be embedded or incorporated within the porous support of the material or the carbon may be a layer, filter or other to be used in conjunction with the material. Additionally, a slow release form, such as a dry capsule, pellet, liposome or other, of a reactivating compound and OP reacting compounds such as certain oximes like HI-6 and mono-bisquaternary oximes such as pralidoxime chloride (2-PAM) may be embedded or incorporated within the porous support of the material.
A preferred embodiment of the invention comprises a material wherein ACHE and/or BChE are simultaneously immobilized with OP hydrolases on or within the porous support during synthesis of the material. Preferably, the enzymes are immobilized through covalent linkages. The enzymes may be of prokaryotic or eukaryotic origin. The enzymes may be contained within the cell or cell free. The enzymes may be of recombinant origin. Other enzymes capable of hydrolyzing hazardous chemicals such as OP compounds may be employed, for example laccase. Additionally, other OP hydrolyzing enzymes would ensure rapid and complete destruction of any toxic intermediate (for example, phosphoryloximes) that might be generated during the decontaminaiton process. Likewise, enzymes such as triesterase may be used for the decontamination of pesticides in a similar manner as herein described. Preferred enzymes are those that may be reactivated.
The materials of the invention may be placed in containers to complete decontamination the OP compounds on the materials.
In another embodiment, the invention relates to the process of making a material, for the removal, decontamination or neutralization of hazardous chemicals such as OP compounds, comprising a mixture of enzymes immobilized on a porous support. In this embodiment, a mixture of enzymes and a prepolymer are gently and evenly mixed together with minimal degradation of the biotype component so that the resulting immobilized enzyme may effectively decontaminate, neutralize or detoxify an amount of an OP compound. The device utilized folds the components into one another. This is a low shear process. During synthesis of the material by prior art methods, for example a mixing drill, the enzymes utilized are subjected to fluid forces or shear stress. Use of a device that gently folds the components into one another greatly reduces these fluid forces or shear stress, and is the preferred device for enzymes, specifically enzymes that are sensitive to the high shear forces of the drill mixing device. Additionally use of additives such as surface-acting polymers, e.g. P-65, or low concentrations of glycerol protects against enzyme denaturation induced by shear forces, and modifies the final properties of the sponge to obtain the desired porosity and absorbent qualities.
In a preferred process of making the material, a two chamber apparatus is utilized. See FIG. 11A and B. One chamber contains a mixture of enzymes and the other chamber contains the prepolymer. The mixture of enzymes and the prepolymer are simultaneously extruded at a 1:1 ratio and mixed. Preferably, the mixture of enzymes and the prepolymer are rapidly and evenly extruded through a static mixing stator which gently and evenly mixes the enzymes and prepolymer. A preferred low shear device is a double chamber syringe and a static mixing stator typically used to mix viscous polyurethanes or epoxy glues. The size of the apparatus may vary depending on need. It may be pocketsize for use in the field by soldiers. Alternatively, the apparatus may be suitable for large-scale production and/or decontamination of a large objects or area. The low shear mixing device more than doubles the resultant ACHE or BChE immobilized enzyme activity when compared to an identical mixture prepared with the high shear device.
The invention further relates to various materials, methods and devices for reactivating the enzymatic activity of the material. These materials, methods and devices will allow a person to use the decontamination material of the invention for several separate uses and/or for a single and continuous use, which would normally require several decontamination materials but for reactivation of the enzymatic activity of the immobilized enzymes. Additionally, these materials, methods and devices allow for complete decontamination and/or neutralization of excess OP compounds absorbed by the porous support but did not react with the immobilized enzymes. These methods and reactivation materials employ substrates and/or oyimes, to reactivate the catalytic activity of the OP inhibited and immobilized enzymes.
The invention further relates to various materials and additives that are added to the embodiment to aid in the removal and decontamination of organophosphate from surfaces such as cracks, crevices, porous or uneven surfaces such as clothes and biological surfaces that readily absorb the organophosphates or pesticides such as skin. The additives are used in conjunction with the sponge material and may be incorporated within the porous support of the material. The additives may be in a dry or liquid form, and may be organophosphate solubilizing compounds such as triacetin or tetraglyme, or oximes, which both aid in decontaminating and reactivating enzymes.
Another embodiment of the invention relates to a variety of kits. Along with the sponge containing a plurality of enzymes needed for the decontamination of organophosphorous and/or sulfur compounds the kit may include materials which would facilitate or be deemed necessary for the decontamination process. Kits may also include polymeric materials and enzymes if the foam is transient in nature, e.g. the prepolymer, a stable enzyme mixture and a low shear apparatus for making an organophosphorous and/or organosulfur decontamination foam. The kits may contain items to facilitate the use of the device, e.g. instructions, containers, test tubes, etc.
This invention is further understood by reference to the drawings wherein:
FIG. 1A illustrates the modeled surfaces of acetylcholinesterase, butyrycholinesterase and phosphotriesterase.
FIG. 1B illustrates the modeled surface of laccase.
FIG. 2 shows a cured material.
FIG. 3 schematically illustrates the specific reaction of the enzymes with prepolymer and the product polyurethane crosslinked ChE.
FIG. 4 shows the linear correlation between the amount of BChE added during synthesis of the material and the amount of BCHE in the final material.
FIG. 5 shows the increasing amounts of BSA added during synthesis to a constant amount of ACHE and TDI polymer.
FIGS. 6A and B illustrates that the materials maintained enzymatic stability for more than 12 months at 25� and 45� C. and 3 years at 4� C. for AChE (A) and BChE(B).
FIG. 7 shows that the material maintained enzymatic activity after consecutive washes.
FIG. 8 shows a substrate concentration dependent curve for soluble and polyurethane coupled AChE.
FIG. 9 illustrates the pH range of soluble and immobilized ACHE.
FIG. 10 shows the relative activities of co-immobilized ChEs and OPHs.
FIGS. 11A and B shows a version of a manual mixing gun and a disposable mixing stator.
FIG. 12 illustrates how oximes reactivate alkylphosphorylated ChE.
FIG. 13A illustrates the enzyme activity of immobilized FBS-AChE. FIG. 13B illustrates the enzyme activity of immobilized Eq-BChE.
FIG. 14 represents inhibition of foam-immobilized FBS-AChE by DFP and reactivation by HI-6.
FIG. 15 represents inhibition of foam-immobilized Eq-BChE by DFP and reactivation by TMB4.
FIG. 16 shows restoration of the original sponge activity with HM-6.
FIG. 17 (A, B and C) shows the protection afforded by sponge with various additives, i.e. tetraglyme (A), HI-6 (B) and 2-PAM (C).
FIG. 18 shows the capacity of the resulting carbon sponge for binding methylene blue.
Enzymes have been incorporated in hypo-based urethane foam during polymer synthesis. See U.S. Pat. No. 4,342,834. Hypoprepolymer is synthesized from a reaction of polyether (or polyester) polyol with isocyanates in the presence of cross-linking agents. See Havens, P. L., et al., Ind Eng Chem Res (1993) 32:2254-2258; U.S. Pat. No. 4,137,200; LeJeune, K. E., et al., Biotechnology and Bioengineering (1999) 20;62(6):659-665. Synthesis is initiated by bringing water molecules into contact with isocyanate groups present within the polyurethane prepolymer.
A two-step procedure occurs from this point. Isocyanates react with water to form an unstable carbonic acid, which in turn degrades to an amine yielding CO2 that gives the porous support lift and enables it to rise. The amines readily react with isocyanate groups, leading to production of urea type linkages. Since the enzyme contains multiple functional groups, such as amines and hydroxyls that can react with isocyanates, the enzyme becomes an integral part of the porous support during synthesis. Significant quantities of enzyme can link to the porous support without disrupting the progress of polymer synthesis. The reaction occurring during the polymer synthesis is shown below.
1. CO2 Evolution:
2. Urea Linkage:
3. Amine Group Enzyme Immobilization:
4. Hydroxyl Group Enzyme Immobilization:
The following list of enzymes and chemicals are examples of those suitable for use in the instant invention:
Acetylcholinesterase (AChE);
Butyrylcholinesterase (BChE);
Organophosphate hydrolases (OPH);
Pseudomonas diminuta bacterial OPH (paraoxonase);
Organophosphate acid anhydrase (OPAA)
Pralidoxime chloride (2-PAM);
7-(methoxyphosphinyloxy)-1-methylquinolium iodide (MEPQ);
Diisopropyl fluorophosphate (DFP);
Acetylthiocholine iodide (ATC);
S-butyrylthiocholine iodide (BTC);
5,5′-dithio-bis(2-nitrobenzoic acid) (DTNB);
N,N′-trimethylene bis(pyridinium-4-aldoxime) dibromide (TMB4); and
1-(2-hydroxyiminomethyl-1-pyridinium)-1-(4-carboxyaminopyridinium)-dimethylether hydrochloride (HI-6).
The following examples are intended to illustrate but not to limit the invention.
As polyether prepolymer derived from tolyl diisocyanate (TDI), reacts most favorably with free aliphatic amines such as lysine and arginine present on the surface of the ChEs (or any protein) to become a permanent cross-linked part of the material, computer aided molecular modeling of the enzymes was performed to highlight the available amino groups on the surface of each enzyme, and to determine whether the coupling of these groups to a porous support would interfere with enzymatic function. This may be performed on every enzyme for which its crystal structure is known, or enzymes which may be modeled by homology.
FIG. 1A illustrates the modeled surfaces of acetylcholinesterase, butyrycholinesterase and phosphotriesterase and shows the lysine and arginine residues on the surface of the ChEs which are available for coupling to the prepolymer. This was generated by Insight II, molecular modeling software, by Biosym Technologies. Based on the molecular modeling, there are at least one lysine and 29 arginine water-accessible residues on the surface of FBS-ACHE to couple to the porous support, while 26 lysine and 26 arginine residues were modeled for equine-BChE. The majority of the lysine and arginine residues were found on the backside of the ChEs, and only a few are found on the side of the enzyme where the catalytic site gorge is located. The rim and the catalytic site gorge opening of both AChE and BChE appeared to be essentially devoid of lysine and arginine. Therefore, coupling these enzymes to the porous support should have minimal effect on the entrance of substrate, inhibitors such as OPs, or reactivators such as oximes which includes mono-disquarternary oximes, release of products of catalysis to and from the active site, and the kinetic rates of the enzymes. Similarly, a model of the surface of laccase (FIG. 1B) is shown with available residues to couple covalently to the prepolymer.
A typical synthesis of the material comprises mixing enzymes in phosphate buffer containing 1% (final concentration) surfactant with prepolymer. Polyether prepolymer derived from tolyl diisocyanate (TDI), Hypol prepolymer TDI 3000 (Hampshire Chemical, Lexington, Mass.), and Pluronic P-65 surfactant (BASF Specialty Chemicals, Parsippany, N.J.) were used. The 2-phase system is mixed and placed into a suitable mold and left to cure. FIG. 2 shows a cured material which comprises a sponge-like support.
FIG. 3 schematically illustrates the specific reaction of the enzymes with prepolymer. Synthesis begins when H2O molecules react with the isocyanate groups present within the polyurethane prepolymer. Isocyanate reacts with the water to form an unstable carbonic acid, which degrades to an amine yielding CO2. The CO2 causes the polymer to rise and become porous, and simultaneously the amines readily react with the isocyanate groups leading to urea linkages. While the amino groups are the preferred reaction site between the enzyme(s) and the prepolymer, hydroxyl (OH) groups are also available to interact within the reaction buffer, e.g. H2O and on the enzyme(s).
Since the ChE contains amines that are on the surface and available to react with the isocyanate groups, they can become an integral part of the polyurethane support during synthesis. There is no significant entrapment of the enzyme in the material as found with cyclodextrins, or physical adsorption of the enzymes, as observed with activated carbon. The inclusion of a surfactant such as Pluronic P-65 at about 1% final concentration controls the final structure and absorption potential of the material.
To create a material comprising a porous polyurethane support, approximately 30 mL of 50 mM phosphate buffer, pH 8.0, containing P-65 surfactant buffer, was placed in a 600 mL plastic beaker. 3 to 5 mL of either purified FBS-AChE (7500 units) or purified Eq-BChE (5000 units) was added, followed by approximately 40 gm of Hypo 3000 prepolymer (tolyl diisocyanate). The two-phase system was mixed and the material was allowed to expand for 10 min, extruded from the container. The material was washed thoroughly with 50 mM phosphate buffer, pH 8.0, dried and stored in a zippered bag at 4� C. for future use.
Approximately 20-90% of the enzymes were covalently linked to the porous support through free amino- or hydroxyl groups. This was determined by the presence of enzyme in first and second washes of the material.
Since the enzymes can be attached at multiple points, they become a part of the cross-linked polymer support. The cross-linked polymer support imparts considerable stability to the bound enzymes. A large quantity of enzyme can be incorporated into a small polyurethane support, thereby rendering the cross-linked polymer support a highly effective material for decontamination.
A. Enzymatic Activity.
Five samples of materials containing FBS-ACHE and five samples of materials containing Eq-BCHE, ranging in weight from I to 40 mg, were suspended in 2.8 mL of 50 mM phosphate buffer, pH 8.0, and assayed using the method of Eliman. See Ellman, G. L., et al., (1961) Biochem Pharmacol. 7:88-95. A linear correlation was found between the weight of the sponge and enzyme activity for both FBS-ACHE and Eq-BChE immobilizations. See FIGS. 13A and B. The linear correlation between the weight of the material and enzyme activity indicates a uniform immobilization of ACHE or BCHE throughout the material.
The material was washed with either 50 mM phosphate buffer, distilled water, or 10 mM ammonium bicarbonate without affecting substrate hydrolysis. Therefore, the mixing of prepolymer, surfactant, and enzyme in situ at 22� C. yields a useful and effective material retaining about 50% of the original activity of soluble ChE.
B. Protein Loading Capacity.
The material has a significantly higher loading capacity for ChEs such as BCHE or ACHE. The final activity of the BChE immobilized in the material could be increased by adding larger quantities of enzyme during synthesis. See FIG. 4. When nonspecific protein (bovine serum albumin, BSA) was added to a constant amount of purified ACHE, there was no reduction in ChE activity. See FIG. 5. Thus, higher potency materials may be synthesized with additional proteins, enzymes and other ChEs. Additionally, materials effective against a diverse array of OP compounds may be readily synthesized by with combinations of multiple enzymes or a plurality of enzymes.
C. Enzymatic Stability.
As illustrated by FIG. 6, the immobilized ChE and OP hydrolase maintained enzymatic stability for more than 12 months at 25� C. and 45� C., respectively. If the material is frozen in liquid nitrogen, most of the original activity remains. TDI imparts remarkable stability to the immobilized ChE; about 50% of the original activity of the immobilized ACHE and 20% of the activity of the immobilized BCHE remained after 16 hours at 80� C., conditions under which the soluble enzymes would exhibit no activity. The ChE materials can be exhaustively dried under vacuum at 22� C. and then rehydrated without loss ofenzyme activity. When AChE or BChE materials were exhaustively washed and assayed for activity, the wash and assay cycle repeated more than twenty times over three days, no decrease in activity occurred. See FIG. 7. This indicates that the material may be used repeatedly.
These results also demonstrate that the ChEs are covalently cross-linked in the porous support and that the ChEs will not leach out to skin, water, or equipment. Therefore, once the immobilized enzymes bind an OP compound the OP is removed from the surface requiring decontamination.
D. Kinetic Constants.
The number of active sites of either the immobilized or soluble ChEs was determined by titration with the organophosphorous compound MEPQ, 7-(methylethoxyphosphinyloxy)-1-methylquinolinium iodide. The bimolecular rate constants for the inhibition of AChE material and BCHE material and the respective soluble enzymes by MEPQ at 25� C. showed that there was no significant difference between the soluble and covalently bound enzymes. See Table 1.
These results demonstrate that the immobilized and soluble forms of ChEs interact with the OP compounds similarly. Therefore, enzymatic activity assays which are generally available and known in the art may be used.
An initial rates method using a modified Eliman's assay was used to determine the parameters Km, kcat, and kcat/Km for immobilized and soluble AChE and BChE. The number of active sites of either the coupled or soluble ChEs was determined by titration with MEPQ. As shown in Table 2 and FIG. 8 for ACHE, the Km values for the immobilized ChEs were about 10-fold greater than the corresponding soluble enzymes, and the kcat values were less dramatically affected. The combined effects on affinity for substrate and kcat resulted in approximately a 20 to 50-fold decrease in acylation (kcat/Km). Interestingly, while soluble BChE lacked substrate inhibition, immobilized BChE yielded substrate inhibition. These results suggest that covalent binding of surface residues of ChEs to the porous support changed some properties of the active site region of the bound enzymes directly or indirectly.
Generally, immobilized cholinesterases or OP hydrolyzing enzymes exhibit between the same to 10 fold greater Km values than the corresponding soluble enzymes. In addition to the cholinesterases, OPH (derived from Pseudomonas diminuta, FIG. 19A) shows about a 10-fold increase in Km because a shift to the right is also observed in the immobilized (sponge) form when determined using the substrate paraoxon. On the other hand, OPAA (derived from Alteromonas, FIG. 19B), shows little change in Km for the substrate paraoxon.
E. pH of Soluble and Immobilized Enzymes.
The pH profiles of immobilized and soluble ACHE are identical and the enzymes exhibit activity throughout the broad pH range of 7-8.5. See FIG. 9. Since the pH profiles of soluble cholinesterases and OP hydrolases have optimal activities in this same pH range, the materials may be optimized and diversified by employing a plurality of these multiple enzymes immobilized on or within a porous support.
ChEs were co-immobilized with bacterial OP hydrolase (OPHB) and/or rabbit serum OP hydrolase (OPHR). There was no reduction in the enzymatic activities of AChE or BChE co-immobilized with OPH as compared to the enzymatic activities of each of these enzymes individually immobilized. See FIG. 10. Additionally, there was no reduction in the enzymatic activity of co-immobilized OPH. Therefore, a plurality of enzymes, which each enzyme differentially reacts with various OP compounds, may be selected and utilized in a material to create a decontamination material effective against a wide range of OP compounds.
By utilizing a method of syntheses modified from the adhesive industry (CPA, Greenville, R.I. 02828) shear forces which decrease enzymatic activity are reduced. See FIG. 11. In this method, the enzyme is not in an organic buffer as required in some immobilization techniques. This results in less air-induced shearing, thereby maintaining enzymatic activity. This method is also simple to conduct, rapid and reproducible. The low shear mixing device more than doubles the resultant AChE and/or BChE immobilized enzyme activity when compared to an identical mixture prepared with the high shear device such as a mixing drill. See Table 3.
100 mg samples of immobilized FBS-AChE were incubated with varying concentrations of DFP in 2 mL of 50 mM phosphate buffer, pH 8.0, for 1 hour at 25� C. In parallel experiments, 1 mM HI-6 was added to the same amount of material and DFP. Residual DFP in the samples was measured by adding a 0.5 mL aliquot of the reaction mixture to 0.5 mL of a fresh 1 U/mL solution of FBS-AChE, incubating for 1 hour, and assaying 10 T1 aliquots using the Ellman procedure. The results are shown in FIG. 14.
The inhibition of FBS-AChE activity by DFP was proportional to the stoichiometric amount of DFP added to the foam suspended in buffer. The presence of 1 mM HI-6 nearly completely prevented enzyme inhibition by DFP. This indicates that immobilized FBS-AChE may be repeatedly reused after reactivating the enzyme with an oxime solution such as HI-6.
FIG. 12 illustrates how oximes may reactivate alkylphosphorylated ChE activity.
50 mg samples of immobilized Eq-BChE were incubated with varying concentrations of DFP in 2 mL of 50 mM phosphate buffer, pH 8.0, for 18 hours at 25� C. In parallel experiments, 1 mM TMB4 was added to the same amount of material and DFP. Residual DFP in the samples was determined by adding a 0.5 mL aliquot of the reaction mixture to 0.5 mL of a fresh 1 U/mL solution of Eq-BChE, incubating for 1 hour, and assaying 10T1 aliquots using the Eliman procedure.
TMB4 was used as a reactivator instead of HI-6, since TMB4 is a more efficient reactivator of inhibited Eq-BChE than is HI-6. These results are shown in FIG. 15. As in Example 6, the foam-bound Eq-BChE may be repeatedly reused after reactivating the enzyme with an oxime solution such as TMB4.
50 mg samples of immobilized acetylcholinesterase were incubated with varying concentrations of MEPQ in 2 mL of 50 mM phosphate buffer, pH 8.0 at 25� C. for 1 hr. In the absence of oxime HI-6, the sponge soaks up the MEPQ and is inactivated. Addition of HI-6 reactivates the sponge's activity, and the MEPQ is detoxified, and most of the original activity of the sponge returns. Only at very high ratios of organophosphate (1000-fold molar excess) is the process of binding, reactivation, and detoxification not complete. However, fresh HI-6 can restore most of the original activity once again. See FIG. 16.
Sponges approximately 1��2�″ (H�L�D) contained 9.0 mL of additive and a second sponge contained 4.5 mL of additive. Each guinea pig was wiped with the first sponge and then the second sponge after soman (GD) exposure. Survival of the guinea pigs was determined after 24 hours, and the protective ratio determined. The protective ratio is the ratio of the LD50 of the sponge containing an additive to the LD50 of soman in the absence of sponge. Thus, the higher the LD50, then the higher the protective ratio and the more effective the sponge combination is for decontamination of guinea pig skin and protecting the animal from the organophosphate. The sponge was compared to the M291 kit (available from Truetech, Inc.), the currently used decontamination kit fielded by the US Army. As shown in the table, the sponges provide 4 to 5-fold better protection than the M291 kit.
FIG. 17A shows the protection afforded by tetraglyme; FIG. 17B the protection afforded by HI-6, and FIG. 17C the protection afforded by 2-PAM. The number on the top of each bar shows the number of guinea pigs evaluated at the indicated dose of soman (GD). For reference, the LD50 of soman on guinea pigs without any effort to decontaminate is shown by the label "GP", while the protection offered by the M291 kit is shown by "M291". Other additives to the sponge such as triacetin also afforded some additional protection compared to the M291 kit.
0.5-1 grams of activated carbon was added to about 4 mL of the prepolymer prior to mixing with acetylcholinesterase (5 mL of Electric eel, in 50 mM pH 8.0 phosphate buffer with 1% Pluronic P-65) to produce an acetylcholinesterase immobilized carbon sponge. The addition of carbon did not interfere with the immobilization of the enzyme, as shown in the table. The capacity of the resulting carbon sponge for binding methylene blue (an colorimetric indicator for 1) activated carbon or resin) is illustrated in the FIG. 18. Therefore, comparison of the sponge with activated carbon to the sponge lacking activated carbon demonstrates that it can bind about 2-fold moremethylene blue at less than saturating concentrations.
To the extent necessary to understand or complete the disclosure of the present invention, all publications, patents, and patent applications mentioned herein are expressly incorporated by reference therein to the same extent as though each were individually so incorporated.
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AA Importing Oriental Landscape 2 Drawer Chest
AA Importing Oriental Landscape 2 Drawer Chest
Weights & Dimensions
|Product Type: Accent chest
|Base Finish: Black
|Distressed: No
|Hand Painted: Yes
|Shape: Rectangular
|Country of Manufacture: China
About the ManufacturerA.A. Importing Company, Inc. was founded in 1934 by A.A. Gralnick in the Gas Light Square area in St. Louis, MO. It began as a distributor for kerosene lamp parts and accessories. Prior to and after World War II, A.A. Importing was heavily involved in the importation of fine crystal and art glass, as well as fine porcelain from Europe. The company became famous for its fine selection of art glass and accessories for the antique trader as well as replacement parts. That was the beginning of building a strong foundation for A.A. Importing in the antique reproduction business. As the nation developed an even greater interest in decorating with an antique theme, A.A. Importing was again at the forefront. Today their sources of antique reproductions have expanded worldwide.
AA Importing Oriental Landscape 2 Drawer Chest
Customer Reviews
|5 star
|4 star
|3 star
|2 star
|1 star
Nicely made
vivian from New York, NY – Verified Buyer
While this is a nice piece, it is not as colorful as shown in the picture. It is much darker and the colors are muted. I must admit to being disappointed because of the actual dark coloration. It came well packaged and timely and is well made. Cons: much darker and muted than depicted in the photo
Great product that matches the description!
Timothy from Phoenixville, PA – Verified Buyer
We purchased several Oriental Landscape 2 Drawer Chests and it is a great product that matches the description! Pros: Great value and no assembly required!
OUR SKU # AAI1450Oriental Landscape 2 Drawer Chest is also sometimes listed under the following manufacturer numbers: 47838

subset: commoncrawl


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Typography for Lawyers: Essential Tools for Polished and Persuasive Documents
From the Publisher. Based on the popular website, Typography for Lawyers is the first guide to the essentials of typography aimed specifically at lawyers. Author Matthew Butterick, a Harvard-trained typographer and practicing attorney, dispels the myth that legal documents are incompatible with excellent typography. Butterick explains how to get professional results with the tools you already have—quickly and easily. Topics include special keyboard characters, line length, point size, font choice, headings, and hyphenation. The book also includes tutorials on specific types of documents like résumés, research memos, and motions.
For your lawyer friend (we may not like to admit it, but we all have one at least):
As it says on the back cover: Good typography is part of good lawyering. Nobody has ever counted how many arguments or court cases are lost because of bad documents that were difficult to decipher, complicated language notwithstanding. I wonder whether documents are often impenetrable and impossible to understand because the author doesn't want the other party to know what they're up to – that is certainly the case with contractual documents. They need an expert to read, creating work for the legal trade. If a book works for lawyers, it'll work for anybody who writes, edits and produces complex documents.
Matthew Butterick practices civil litigation in Los Angeles but has a background in design and typography. He has just released a typeface designed specially for these types of documents called Equity and has been running the website www.typographyforlawyers.com since 2008.
This is a pleasantly small but comprehensive book, written in a style that betrays Matthew's background in communication. If your lawyer friend doesn't go on to produce better documents after reading this book, it may be time for a new friend (released November 2010).
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Design Books to Win, April 22, 2015: Winner of The New York City Transit Authority Graphics Standards Manual Announced
The winner of The New York City Transit Authority Graphics Standards Manual designed by Massimo Vignelli and Bob Noorda is Joseph Swerdlin — congratulations!
Our new giveaways continue in 2015.
New Book Available for Pre-Order Now; Release Date, June 1, 2015: Spin: 360°
A portrait of Spin, one of London's leading design studios, which has produced work in identity, print, moving image, retail, digital, and environmental graphics. Includes essays and interviews with Spin's founders, Tony Brook and Patricia Finegan, and texts by Paula Scher, Stefan Sagmeister, Ben Bos, Wim Crouwel, Rick Poynor, Steven Heller, Patrick Burgoyne, and artist and author Edmund de Waal.
Tony Brook and Patricia Finegan; designed by Spin
Published by Unit Editions
Pre-order and details. The first 1,000 copies come with a limited-edition pack of six silk-screened cards in a matching envelope, plus a set of six button badges — designed by Spin. If you pre-order you also receive an exclusive advance excerpt from the book.
New Book Release, February 9, 2015: Reproducing Scholten & Baijings
The first book on the work of the Amsterdam-based studio Scholten & Baijings, which has become renowned for its sensitive and subtle yet functional products—from ceramics and silverware to textiles and even a concept car.
By Louise Schouwenberg
Published by Phaidon Press
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New Book Release, December 5, 2014: Dream of Venice
Captures the mysterious allure of the ancient floating city with the evocative photography of Charles Christopher and the beguiling words of a diverse group of contemporary Venetophiles.
Edited by JoAnn Locktov; photography by Charles Christopher; foreword by Frances Mayes
Published by Bella Figura Publications
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New Book Release, February 10, 2015: Design to Grow: How Coca-Cola Learned to Combine Scale and Agility (and How You Can Too)
A Coca-Cola senior executive shares both the successes and failures of one of the world's largest companies as it learns to use design to be both agile and big. In this rare and unprecedented behind-the-scenes look, David Butler and senior Fast Company editor, Linda Tischler, use plain language and easy-to-understand case studies to show how this works at Coca-Cola—and how other companies can use the same approach to grow their business.
By David Butler and Linda Tischler
Published by Simon & Schuster
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A North Hardin High School student has died the day after what should have been his graduation day.
Senior Thomas Carmen died at 2:40 Sunday afternoon, Vine Grove Police Chief Dale Riggs said, after sustaining injuries May 12 in a car wreck.
Carmen was a passenger in a vehicle that appeared to have spun out on Crume Road in Vine Grove because of slick road conditions. The car turned about 165 degrees and hit the front of an oncoming vehicle with its back end.
Hooray for Heroes thanked uniformed men and women this weekend for their service.
The seventh annual Hooray for Heroes was Saturday at Stithton Baptist Church in Radcliff. Free food and fun were offered and many families took advantage. The festival honors the armed forces and emergency responders.
Sgt. Maj. Luke Legg was working a grill at the festival, flipping burgers on behalf of the Bluegrass Sergeants Major Association. The association has worked with the event every year, but this was Legg's first year working.
Those who have paid the ultimate price for their work were remembered Friday.
The William R. Burns Memorial Fraternal Order of Police Lodge 39 hosted a Law Enforcement Memorial Service at the Radcliff Police Department, to remember those police officers who have died in the line of duty and to coincide with National Police Week.
Wayne Fleming, vice president of the lodge, led the proceedings and said before the event the memorial is "to honor and remember those who gave the ultimate sacrifice." The lodge itself is named after one such officer.
Clients at SpringHaven domestic violence shelter in Elizabethtown were filtering back to the facility Friday after they were transplanted to hotels a week before when a state inspector found a bridge leading to the facility was unsafe for large vehicles.
Tanya Thomas, assistant director of SpringHaven, said the shelter got word to evacuate around 21 clients on May 11 after an inspection found the bridge could not safely sustain weights exceeding three tons.
Motorists who attempt to fake out police regarding seat belts soon will be under even more scrutiny.
Local law enforcement agencies and Kentucky State Police will be participating in the national Click It or Ticket campaign starting Monday and ending June 3. The directive is aimed at reducing the number of highway fatalities through an increased focus on the enforcement of seat belt laws.
Fans of the Beatles can relive the band's glory Monday at the Hardin County Public Library's BeatleMania.
The celebration begins at 6 p.m. in Elizabethtown and features a documentary, a variety of games with prizes, and of course, music from John, Paul, George and Ringo.
The event celebrates the 50th anniversary of Beatlemania, according to Kathy Shireman, organizer of the celebration and circulation worker at the library.
Tuesday's primary likely will yield a low voter turnout.
Because large swaths of the county only have an already decided presidential primary to vote on, Hardin County Clerk Kenny Tabb said he would not be shocked if the turnout is as low as 10 percent.
Two inmates who walked away from the restricted custody area of the Hardin County Detention Center early Friday morning were captured riding bikes early Sunday morning in Radcliff on U.S. 31W.
Brandon Board, 21, of Bardstown, and Vance Tate, 37, of Radcliff, were captured at 12:49 a.m. Sunday by Radcliff Police after a short foot chase. The two were taken in without incident, according to an RPD news release.
A Radcliff teenager who appeared to thwart cancer in March once again is battling for his life.
Andrew Holland, diagnosed with a rare form of Ewing's sarcoma of the spine and who was Radcliff's junior mayor in April 2011, is back in treatment following the cancer's aggressive return.
Holland's mother, Regina Hensley, said the 15-year-old aspiring writer's condition has worsened considerably with the emergence of nine cancerous tumors – seven on his spine, one on his liver and another on his brain.
Copyright © 2015
The News-Enterprise is your source for local news, sports, events and information in Elizabethtown, KY, Hardin County and the surrounding area.


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Made of 100% cotton twill with 100% polyester laminated buckram, this one of a kind cap is a great fit for your business! This six-panel, low-profile structured cap comes with an adjustable velcro strap and black trim with white piping. Our overseas direct program delivers big savings! Custom colors may be available depending on quantity. See our website for current pricing. Lifetime guaranteed. This supplier is QCA certified. One size fits all.
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My name is Frankie. I'm a happy-go-luck bunny boy who is playful and curious. I am mellow and loving. I'd make a super sweet companion bunny for your or your girl bunny who is looking for a husbun. Here's me showing off at an adoption event at Christmas time. I look good in red, eh. Don't overlook me because I'm white with ruby eyes. I'm all personality!
"Bunnies Gone Bad: The 7 Deadly Sins" is a wickedly funny and amazingly realistic portrait of the rabbit psyche.
Two each of seven for set of 14 cards representing each sin. Comes with seven bright green envelopes as well. Cards measure 5", by 7".


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You want something creative to happen…so contact us. It's as simple as that.
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SeafollyBeach Crush Bandit Dress
In a cute see-through design, the Seafolly Beach Crush Bandit Dress is the perfect cover-up for the beach. Made from 100% cotton lace, this dress is sure to become a holiday favourite. Please note that the slip underneath is not included.
Product code: SF-52191
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Originally written on Fox Sports | Last updated 8/11/12
Jeff Wolfert kicked a 45-yard field goal with 28 seconds left to lift the Cleveland Browns to a 19-17 win over the Detroit Lions on Friday night. Detroit stopped Adonis Thomas on a 2-point conversion run with 6:05 left to keep the lead, but couldn't run enough clock to keep the ball away from the Browns. The Lions' last shot ended when rookie Kellen Moore was intercepted by David Sims at the Cleveland 36 with 34 seconds left. The banged-up Browns took another hit when rookie Brandon Weeden threw to Mohamed Massaquoi and the wide receiver left the game with a head injury. Weeden was 3 of 9 for 62 yards with an interception and a fumble. Detroit backup quarterback Shaun Hill led two touchdown drives in the first half to put the Lions ahead 14-3 in the first half. When the reserves took over in the second half, Cleveland outscored the Lions 16-3. Hill set up Keiland Williams 5-yard TD late in the first quarter with a 27-yard pass to Maurice Stovall. The backup quarterback connect...
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Vubiquity Acquires FilmFlex
Vubiquity, the leading provider of multiplatform video services, today announced that it had acquired UK-based FilmFlex Movies Ltd., one of the largest VOD and multiplatform providers outside the United States. FilmFlex, formerly a joint venture between Sony Pictures Television and The Walt Disney Company Limited, provides digital storefront and video on demand services to connected consumers throughout the UK. The company has licensing agreements with major US studios and many independent distributors.
With over 90 million transactions enabled since launch, FilmFlex is one of the largest independent providers of white label VOD services outside the United States. FilmFlex offers a customizable digital video storefront, already used by service providers and brands such as Virgin Media, TalkTalk, Film4 and EE, as well as content aggregation and delivery to set-tops and connected devices. Bringing the two companies together extends Vubiquity's relationships with major global content and service providers and adds new licensing and digital storefront capabilities to Vubiquity's offerings.
"Vubiquity remains focused on expanding the breadth of our managed services and technical solutions that today support content and service providers worldwide," said Darcy Antonellis, CEO of Vubiquity. "With FilmFlex we add additional studio assets and licensing, and gain important front-end technology to enable EST and other monetization models for multiplatform video consumption."
Vubiquity's acquisition of FilmFlex follows the launch of the company's AnyVU Cloud platform, which offers a broad set of modular services that include a deep library of licensed content for both linear and on-demand, set-top box delivery and direct-to-subscriber streaming, electronic sell-through (EST) as well as other next-generation video services. FilmFlex's digital storefront, assets and capabilities will be integrated into the new platform, and offered as part of the suite of modular services available to Vubiquity's customers.
"With the addition of FilmFlex's proven, carrier-class digital storefront to the AnyVU Cloud platform, Vubiquity offers a complete end-to-end solution for quickly launching and monetizing multiplatform digital video services, while also allowing service providers who prefer a modular approach to choose only those services that fit in with their strategic plans," continued Antonellis.
"FilmFlex's assets and capabilities, when merged with Vubiquity's reach and scale, result in a very large opportunity for both companies -and for the customers we share," said Jeff Henry, FilmFlex's CEO. "We look forward to continued growth together as we integrate our companies."
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- CONSTELLATIONS' Jake Gyllenhaal and Ruth Wilson Set for THEATER TALK This Weekend


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Goodreads helps you keep track of books you want to read.
Start by marking "The Letters To Seven Churches: Updated Edition" as Want to Read:
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Preview — The Letters To Seven Churches by William Mitchell Ramsay
The Letters To Seven Churches: Updated Edition
This edited and updated edition of Ramsay's classic work has, except for the omission of a lengthy poem in chapter 14, left the original material intact. The language and style were updated for the modern reader, modern place names were used, and a modern translation is the source of scripture citations. Notes especially have been updated and corrected, and fuller bibliogr ...more
Published November 1st 1994 by Hendrickson Publishers
(first published 1904)
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Sir William Mitchell Ramsay was a Scottish archaeologist and New Testament scholar. He was the first Professor of Classical Archaeology at Oxford University and pioneered the study of antiquity in what is today western Turkey.More about William Mitchell Ramsay...
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Frozen Watermelon Slush
A refreshing, colorful drink for those dog days of summer. If you have extra watermelon, you can freeze it for more slushes.
Total Time:
Level: Moderate
Yield: 3 1/2 cups
Advertisement - Continue Reading Below
- 3 c. seedless watermelon
- 1 c. frozen strawberries
- 2 scoop lemon sorbet
- 0.25 c. pineapple juice
- 2 tbsp. fresh lime juice
- In blender, combine watermelon, strawberries, sorbet, pineapple juice, and lime juice and blend until mixture is smooth. Pour into 4 glasses.
What do you think?
How to Make a Liberty Bell Cake
Let freedom ring with this fun party centerpiece.
How to Make an Uncle Sam Hat Cake
Because America's birthday deserves a little something special.
Wish Bone Recalls Ranch Dressing Due to a Dangerous Mistake
Double-check your dressing, everyone.
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Town Square
Athertonian settles at $5,500 after losing $160 million
Original post made by anonymous, Menlo Park: Suburban Park/Lorelei Manor/Flood Park Triangle, on Nov 17, 2010
For someone with a reputation for brilliant decisions, why didn't she see this coming in advance and pay her housekeeper the nine grand in back wages? Seems that this dispute was the nail in her political coffin.
She was running neck and neck with a political retread until California found out about her illegal employee of nine years. Seems her consultants wanted to air this out in the primaries, but it was Meg's decision to cover it up and hope no one found out. It's always the coverup in politics, isn't it?
Getting chintzy on nine grand, after spending $160 million on ads and consultants.
Comments (2)
a resident of Woodside: other
on Nov 18, 2010 at 7:15 pm
Stunning that she thought an election could be overpowered by sheer volume of cash, yet nickel and dime'd the person who was her undoing.
a resident of another community
on Nov 23, 2010 at 1:58 pm
Meg gets vindication that the working families she rails against were in fact against her: nurses union assisted Nikki Diaz.
"According to an account published in the San Francisco Chronicle on Tuesday, the California Nurses Assn. helped Diaz get in touch with Allred. The nurses union had been a thorn in Whitman's side throughout the campaign, protesting at events and dressing up an actress as "Queen Meg.""
Meg, dear, you should have actually listened to your highly paid advisers, when they told you not to cover-up your employing the illegal.
It's always the cover up.
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A suspenseful new teaser for the new Star Wars film 'The Force Awakens' debuted during the Star Wars Celebration today. Watch it here…
Remembering Andrew Breitbart 3 Years Later #War
Lee Hernly video No Comments andrew breitbart, Conservatism
We miss you terribly Andrew and continue the fight in your absence.
Sesame Street's House of Cards Parody 'House of Bricks'
Lee Hernly video No Comments house of cards, netflix, sesame street
Inspired by the Three Little Pigs, this spoof by Sesame Street of the hit Netflix show 'House of Cards' is glorious. Enjoy!
Coming Soon: The Department of the Internet
Lee Hernly video No Comments fcc, net neutrality
Great video via Protect Internet Freedom. Help save the internet today! The FCC vote on regulations the FCC won't let you or I see happens TOMORROW, Feb 26.
Carol Costello – 'Right Wing Citizen Extremists' More Dangerous Than ISIS
Lee Hernly video No Comments carol costello, cnn, news
Carol Costello says right wing extremists are more dangerous than ISIS. Bet she can't name one.
Joe Biden – Middle Class is Being Killed
Lee Hernly video No Comments joe biden, speeches
Joe Biden says middle class is in bad shape. What have President Obama and Vice President Biden done to help the middle class? Nothing.
Joe Biden – The Hands-On Vice President
Lee Hernly video No Comments joe biden, vice president, white house
Here's a classic compilation of Vice President Joe Biden being touchy-feely with an assortment of women and children.


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Stuffing a big block V8 into just about anything is guaranteed to make for an entertaining ride, but when it's a 1969 Camaro and... More
Around the Streetside showroom, we hear a lot of conversations, and those we hear about this 1968 Camaro lead us to believe that it... More
Think the replica business is a new development? Guess again! This neat little Bugatti replica dates to the 1970s and uses a sturdy and... More
Purists, this may not be your cup of tea, but if you're a guy who loves the old-school Corvette look with all the modern... More
You could probably call this 1968 Chevrolet Chevelle Malibu a sleeper, thanks to its low-profile paint, minimal flash, and big, honking stroker motor under... More
Descended from greatness, this 1972 Plymouth Road Runner means affordable muscle, complete with a strong-running small block and an awesome Mopar horsepower look. Extensively... More
Is it just me or are Oldsmobiles getting more popular? We can't seem to keep cars like this 1964 Oldsmobile F85 hardtop in stock.... More
No, your eyes aren't deceiving you this really is a TON of truck for the money. For that kind of money, you don't usually... More
This 1969 Chevrolet Nova has a lot of things we like. One, the power-to-weight ratio is awesome, particularly with a 400 cubic inch small... More
Here at Streetside Classics, we do love our Oldsmobiles. Plenty of performance with a healthy dose of luxury on the side, plus some of... More
Recently we've featured quite a few Toyota FJ40 Land Cruisers and a few of the British Land Rovers, but we're guessing that you've never... More
OK, we know a lot of you guys out there are picky about your cars, but please take a moment to look at this... More
Finished in its original code 24 Light Blue paint, this gorgeous 1973 Camaro LT offers big performance in one of the best-looking F-body packages... More
Why spend $50,000 on a new truck when awesome haulers like this 1979 Ford F-100 Ranger are still around? Not quite a full-fledged collectable,... More
Mustang fans, you're in luck. This 1970 Mach 1 is a bargain compared to its big block siblings, so if you're looking for a... More


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Insight newsletterMenu
Insight newsletter
Welcome to "Insight - Networks Perform™". This newsletter digs into today's hot mobile broadband topics, bringing expert insight into today's and future technology and business trends to help operators achieve their goals.
Press releases
Telecom industry and European academia join forces to develop a multiservice mobile network architecture for the 5G era


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Billboard Queen
Since first topping the Billboard Hot 100 singles chart in 2006 with "S.O.S.," Rihanna has made a habit of hitting the No. 1 spot with an unmatched string of addictive pop hits.
With the Roc Nation diva unveiling her newest creation, an assist from the great Paul McCartney and Kanye West called "FourFiveSeconds," it has us wondering, will it keep her atop the charts? Check out Rih's other No. 1s.
(Photo: Westbury Road Entertainment/Roc Nation)
Songs That Take a Stand Against Domestic Violence07/06/2015
I Get Money: 50 Cent's Most Powerful Moves07/06/2015
R & Beef07/06/2015
Instagram Photos of the Week: Independence Day Edition07/05/2015
#Freedom: A Social Media Playlist07/04/2015
Out and About Wireless Festival 201507/03/2015
Tweets Of The Week: President Obama Keeps OutKast Rocking07/03/2015
4th of July Turn-Up Mix07/02/2015
Rihanna Proves She Can Be Deadly in 'BBHMM' Video07/02/2015
O Canada!: Canadian Born Singers07/01/2015
Creative Ice Cream Flavor Day07/01/2015
Family Reunion Hip Hop Tours06/30/2015
Album Covers Shot by Photographer Jonathan Mannion06/29/2015
The Rundown: Meek Mill, Dreams Worth More Than Money06/29/2015
Athletes Who Have Turned to Music06/28/2015
Out and About: Meek Mill's TIDAL X Event06/27/2015
J. Cole's Greatest Give-Back Moments With Fans06/26/2015
Instagram Photos of the Week: #Omeeka Up Close and Personal06/26/2015
Tweets of the Week: Joey Bada$$ Squashes His Tensions With06/26/2015
Picture Me Rollin': West Coast MCs From LA to the Bay06/26/2015
50 Cent: The Trollfather06/25/2015
Don't Label Me: Artists Beefin' With Their Record Labels06/25/2015
25 Things Music Lost Since Michael Jackson Died06/25/2015
The Rundown: Tyga, The Gold Album: 18th Dynasty06/24/2015
Who's Watching: MCs Who've Invaded Your TV Screen06/24/2015
Tidal Waves06/23/2015
Minister Farrakhan and Hip Hop06/23/2015
Class Warfare: When XXL Freshmen Clash06/23/2015
Gentlemen's Club: When Rappers Cover GQ Magazine06/22/2015
Kanye, Future and More Rock Birthday Bash 2006/22/2015
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Your Essential Father's Day 2015 Playlist06/21/2015
Hip Hop Pops and Their Kids06/20/2015
Use of the Confederate Flag in Hip Hop06/20/2015
Tweets of the Week: Meek Mill Revokes Rachel Dolezal's Pass06/19/2015
Music Stars With Success at Different Labels06/18/2015


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Consumer Watchdog gets action after neighbors raise concerns about leaning and broken utility poles
FPL expediting wire transfers to new poles
BOYNTON BEACH, Fla. - Splintered poles and low hanging wires concern John Lennon and his neighbors in the Sun Valley Neighborhood of Boynton Beach.
Lennon said the wires look terrible.
The tangled web of wires belong to three utilities including Florida Power & Light, Comcast, and AT&T and create a chain reaction of problems when there's an issue like a recent auto accident.
"The pole was hit down here, but it snapped up here because there was way too much weight in the high areas," explained Lennon.
Lennon said he's concerned with what might happen if a hurricane hits.
"With one automobile taking out one pole we were without power for eight hours on all media and power for two hours," Lennon explained.
Service was restored, but bundled and frayed wires remain.
"It's an eyesore for our neighborhood," explained Lennon.
In Delray Beach, low hanging wires are causing problems for delivery trucks.
"Every now and then one blows through here that doesn't realize that line is hanging too low and just snags it," explained Delray Florist owner Tom Strongosky.
It disconnected the flower company's phone lines and those for others in the strip mall.
"Without a phone line we have no access to our internet website. We can't receive orders and we can't process any credit cards either," explained Strongosky.
Service is back on but the pole that snapped and cracked is still leaning.
Strongosky said he called the Consumer Watchdog because he's getting the runaround.
After our calls, AT&T and FPL came out to look at the problem. The broken pole was removed, and FPL fixed the leaning pole after AT&T fixed the slack in its lines. That reduced the stress on the FPL pole.
In Boynton Beach, there's progress too.
"We know customers are concerned about the situation and so are we," explained Bill Orlove of FPL.
One utility pole was removed, and there are plans to move the second pole soon.
"When we put in a new pole and transfer the lines to the new pole we need to wait for the other utilities to put their equipment onto our pole and then we can remove the old pole," explained Orlove.
FPL is working to speed up the process.
"We are trying to expedite with our partner utilities to make this go smoother than it has in the past," explained Orlove. "We recently spoke to Comcast and entered into an agreement with them that we will be able to transfer some of their lines onto our poles."
FPL is working to make the transfer of wires to new poles smoother, because it's in the middle of a big project to replace 3,000 old poles with stronger, more hurricane resistant poles. The utility owns 117,000 poles in Palm Beach County and said less than 4 percent of them are double poles.
The utility points out that other utilities also use double poles.
A/C donation after Watchdog exposes ripoff
A Port St. Lucie man will sleep in a cool house for the first time in a year thanks to the generosity of two companies.
State closes Blue Front a 2nd time for rodents
Roaches closed the doors of two restaurants, but when they reopened they didn't want to talk about the pest problem. Plus a rat problem…
Don't let tire trouble delay July Fourth travel
When was the last time you checked your tires? The Consumer Watchdog shows you why you may want to check your tires before you hit the road…


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36 year old woman | Sarasota, FL, USA | Seeking men 29-45 within 50 miles
Excited to meet someone normal! interested in meeting a nice, down to earth man I can talk to with no baggage!! is this you?
I'm a beautiful down to earth girl who enjoys being active, not taking myself or others too serious. I would love to spend time with someone who is loving, kind and most importantly funny!! I'm not interested in playing head games. I say what I mean and mean what I say. I'm extremely honest, and value that quality in anyone I spend time with. A great sense of humor is my favorite quality in a partner. I'm a very social person who loves meeting new people. I'm most passionate about friends, family, and physical activity. I would love to share my time with a quality person, who also likes to exercise, enjoy life. and not take themselves too seriously. Most importantly though, I'm looking for a nice person. Someone who is KIND. Someone who has the capacity to love fully, and be loved that very same way. Serious applicants only please. * P. S. If you have a bunch of stupid tat's and a pit bull no thanks:) just being honest.
Book club , Coffee and conversation , Dining out , Exploring new areas , Nightclubs/Dancing , Watching sports , Wine tasting
Cycling, Golf, Other types of exercise, Weights / Machines, Yoga
I exercise 3-4 times per week.
I have Dogs
England, up north
Good movies, great pizza, reading, current events, dancing
exercise, watch movies, spending time with friend's and family. Doing something unexpected!
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|5'5" (165cms)
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my mother is from england
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A Love Story In 22 Pictures
A picture is worth 1000 words. Keep tissues handy. (Photographs from Tim Dodd Photography.)
All photos copyright Tim Dodd photography. Used with permission.
26. More: The 50 Most Romantic Photographs Of All Time
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- Police in Athens used pepper spray on protesters two days ahead of Greece's major bailout vote.
- A medical helicopter has crashed in Colorado. Three people were injured, according to reports.
- Serena Williams survived an upset at Wimbledon and defeated Heather Watson. She'll play her older sister Venus in the next round of the tournament ?


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Dallas' Mahinmi says Kobe is NBA's most arrogant player
Jul 6, 2011, 10:37 AM EDT
The Lakers are down are down, and people are lining up to kick them. After three straight trips to the finals and two titles, the Los Angeles was unceremoniously swept out of the playoffs in the second round by Dallas. Then the Lakers lost their coach, who was a bigger star than most of the players. There are questions about if they can bounce back.
That's when Dallas' backup center Ian Mahinmi decided to kick Kobe Bryant while he was down.
Mahinmi called Kobe the league's most arrogant player in an interview with French NBA site BasketUSA (via the Express-News Spurs Blog and Hoopshype).
"Kobe is super arrogant but everybody loves him," Mahinmi said. "To me, Kobe is more arrogant (than LeBron James)."
Kobe lives for that kind of fuel, we'll see if he takes it out Mavericks when they play next season (and we'll see if in the quest to prove himself he takes the Lakers out of their offensive flow with his domination of the ball). Most players don't play well angry, Kobe is one of the exceptions. Next season could be interesting out west.
- Chris Ross - Jul 6, 2011 at 7:05 PM
I don't think what Ian Mahinmi is saying is necessarily false. I mean, Kobe is known around the league as one of the most arrogant guys and all Mahinmi did was make his statement public. I'm not sure how relevant it is that he is a 3rd string center. He has the ability to voice his opinion as he is an NBA player so why not just answer the question. No one can answer a question honestly these days. I'm not sure what Ian Mahinmi's getting from the media is fair at this point.
- damp1015 - Jul 6, 2011 at 7:35 PM
It makes me wonder if he would have voiced his opinion if he was not a champion? Hmm? Balls get ballsy when you feel big.
- downtowndanny - Jul 6, 2011 at 10:02 PM
- phillysports266 - Jul 7, 2011 at 12:00 AM
Kobe is the type of arrogant that says, "I refuse to lose to this inferior player", so was Jordan and so are all great players. LeBron is the type of arrogant that says "OMG, I don't get it, I'm soooo much better and I'm still losing?"
- jizzojames - Jul 7, 2011 at 12:42 AM
When I saw the headline of this post I was ready to comment, but seriously.. Who is Ian Mahinmi?!! I respect your opinion but who are you?
- illyistic1 - Jul 7, 2011 at 1:51 PM
Will this goober even get a ring? How would he know how arrogant kobe is when he never gets on the court? Im just saying!
- duhgrate1 - Jul 8, 2011 at 10:38 PM
First and Last time Mahinmi and Kobe are ever mentioned in the same sentence.
- jr10cross - Jul 20, 2011 at 3:48 PM
you have to be somewhat arrogant to put up 63 points in 3 quarters in a game and then 2 weeks later throw up 81 because people rip you about missing your chance at greatness. Kobe's attitude is what makes him what he is, an assassin. I love it. I love to see a guy feel he can defeat anyone in his path and make the game of basketball a war of his own mind and totally delete the idea of thinking that the opponent in front of him is even relevant. It's like being a super hero on the court.
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1973 AMC Javelin w/56,900 miles, said to be actual- Due to age is being sold exempt- 360 Cubic inch V-8 Motor, Automatic Transmission, Power... More
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'71 Gremlin X, Beautifully restored ultra rare Gremlin X, Freshly rebuilt 360cid AMC V-8@300 hp, Close ratio 4 speed transmission and Hurst Shifter, Factory... More


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Pope declares St. Anne Melkite Catholic Church a cathedral
A Melkite Greek Catholic church will be elevated to the title of cathedral in Los Angeles, making it a co-cathedral with the Eparchial Cathedral of the Annunciation in Boston. The Eparchial Bishop of Newton, Nicholas J. Samra, will celebrate the solemn inauguration of the soon-to-be-cathedral, St. Anne Melkite Greek Catholic Church, on Aug. 1 at 4 p.m.
St. Cyril of Jerusalem Parish hosts successful casino night
On April 25, St. Cyril of Jerusalem Parish hosted a casino night, complete with dealers and games of blackjack, roulette and craps. The proceeds of the event went to St. Cyril of Jerusalem School.
Louisville senior honored for service
Louisville High School senior Nadia Gonzalez Barranca is being honored for her long hours of service as an altar server, confirmation leader and eucharistic minister. She is one of the few recipients of the Christian Service Award due to her almost 500 hours of community service during her senior year.
Providence Holy Cross Medical Center launches app
This month, Providence Holy Cross Medical Center has launched a new app to help potential patients quickly access efficient medical care.
Altar server commissioning
Twenty men and women were commissioned to the ministry of Altar Server during the 9 a.m. Mass at St. Bernard's Parish on March 8.
Sacred Heart gala to raise scholarship funds
Sacred Heart High School will host its first gala, "United to Empower," on April 25 at 5 p.m. The evening will raise funds for the Comet Scholarship Fund, a tuition assistance program for students at the all-girls college prep school.
Student artists featured in gallery exhibit
The LA City Hall Bridge Gallery is showcasing the artwork of four Louisville High School female students as part of the exhibit, "Expressions of Strength: An Art Exhibit Celebrating the Strength and Vitality of Los Angeles Youth."
Dr. Bill Creasy at St. John Eudes
St. John Eudes Parish will host a retreat entitled "The Passion of the Christ," given by biblical scholar and former UCLA professor Dr. Bill Creasy on March 28 from 9:30 a.m. to 4:30 p.m.
Author makes a visit to Flintridge Sacred Heart
Award-winning author Jan Ellison returned to her former preparatory school, Flintridge Sacred Heart Academy on Feb. 4 to discuss her writing methods, experience and inspirations with students and staff.
Pastoral councils unite for interregional gathering
Pastoral council members from across the Los Angeles Archdiocese's five regions gathered at Providence High School in Burbank Feb. 21 for the Eighth Annual Interregional Gathering.
The meeting featured addresses by Archbishop José H. Gomez, Bishop Gerald Wilkerson of the San Fernando region, and Father Edward Benioff, director of the Office of New Evangelization.
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Sing a Song of Ireland
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The following information pertains to supervisor training. This covers reasonable suspicion, a situation in which a supervisor may feel it is necessary in order for safety to test an employee for drugs or alcohol in their system while on the job. 1,876 more words
Tags » Employee Testing
When an employee quits – what's left behind?
I've been reading blogs and listening to talk shows. It surprised me that there were so many of them busy discussing quitting one's job.
Quitting a current job is a necessary step to new adventures and personal achievement. 355 more words
How to tell if they DO have a clue.
It's unfortunate that this picture makes us smile. Humor is often a recognition of some outness we can reject, so there must be some truth in the original idea. 223 more words
Test Us!
Sometimes one just has to blatantly spread the word; well, advertise of a sort. I just had to announce we've made changes! Our new name is Accurate U-MAN Hiring. 106 more words


text:
Catskill Craftsmen, Inc. Butcher Block Table
Special Offer:
12 Issues of Sunset Magazine
Included with purchase ($20 value).
Catskill Craftsmen, Inc. Butcher Block Table
Weights & Dimensions
|Product Type: Prep Table; Butcher Block
|Base Material: Wood
|Counter Material: Butcher block
|Solid Wood Construction: Yes
|Style: Contemporary
|Exterior Shelves: No
|Drawers Included: No
|Cabinets Included: No
|Towel Rack: No
|Pot Rack: No
|Spice Rack: No
|Cutting Board: Yes
|Foldable: Yes
|Drop Leaf: Yes
|Trash Bin Compartment: No
|Stools Included: No
|Wheels: No
|Wine Storage: No
|Stemware Storage: No
|Finished Back: Yes
|Weight Capacity: 300 Pounds
|Commercial Use: Yes
|Eco-Friendly: Yes
|Product Care: Oil regularly with mineral oil
|Country of Manufacture: United States
|Assembly Required: Yes
|Additional Parts Required: No
|Product Warranty: Replacement parts available for up to 5 years for the duration of manufacturing.
About the ManufacturerCatskill Craftsmen is the nation's leading manufacturer of domestic hardwood ready-to-assemble kitchen carts, islands, and work centers. Catskill also manufactures gourmet butcher block chopping blocks, cutting boards, hardwood cabinets and furniture, plus book carts and racks. Located in Stamford, New York, Catskill Craftsmen has been producing quality wooden products for over 55 years.
Catskill Craftsmen is located in Stamford, New York, a small rural village nestled in the northern sections of the Catskill Mountains, at the headwaters of both the Schoharie and Delaware rivers.
Catskill Craftsmen began operations in 1948, and has developed over the years, becoming the nation's leading manufacturer of RTA (ready to assemble) hardwood kitchen islands, carts, and work-centers. The company is privately held, and takes great pride in its products and services.
Sixty dedicated employees work hard to insure that our products are delivered rapidly and meet your satisfaction. Catskill's manufactured items are made from naturally self-sustaining, non-endangered North American hardwoods, primarily birch and hard rock maple. Lumber is purchased from area sawmills, then dried, manufactured, and packaged on site. All sawdust, shavings and waste materials generated during the manufacturing process is converted into wood pellet fuel, and used to heat homes. Catskill is proud to say, "our operations generate no wood waste".
Catskill is also a leading manufacturer of exclusive, private label kitchen workcenters for the trade. Their products are described as "high quality at fair prices".
Call us if you have questions about Catskill products. We're here to help!
Catskill Craftsmen, Inc. Butcher Block Table
Customer Reviews
|5 star
|4 star
|3 star
|2 star
|1 star
iDEAL !!!
Gillous from Ellenton, FL – Verified Buyer
This combination prep area/dining table for two is just an ideal piece of furniture. When making pies or baking cakes, one always needs extra counter space, this item fits the bill !! It's unfinished as advertised. I treated mine with tung oil, which polishes out to a beautiful shine !! It's easy to put together. I would recommend this item to a friend !!! Also when not needed, it neatly fold up for easy out of the way storage !!!!
A quality product.
Althea from Blackwood, NJ – Verified Buyer
The fold away table is exactly as described online. It is sturdy and of high quality workmanship. The only issue I had was that some of the washers are tough to get in place while assembling. Otherwise it was easy to put together.
Very Nice, Good Quality
Christian from Accord, NY – Verified Buyer
Got this for our family room to use instead of a couple of flimsy folding tables... And we're very happy with it, serves its purpose and suits our needs just fine.
Handy, sturdy, practical.
HEIKKI from WIMBERLEY, TX – Verified Buyer
Perfect to our small kitchen. Also works for a dining table for grandchildren. Pros: Good value, sturdy, handy. Cons: Surface was not finished with coating. I did the coating and now it is perfect.
Great product, matches description.
Andrea from High Falls, NY – Verified Buyer
This product is exactly as described. The butcher block is sturdy and beautiful. A nice way to share a romantic dinner with your honey.
Butcher Block Table
Colleen from STAMFORD, CT – Verified Buyer
Nice solid table. Great for putting food from oven on for small buffet. Very solid& assembly was easy.
High quality and functional
Rebecca from Seattle, WA – Verified Buyer
This table was easy to assemble and is made of a high quality of wood. The table is great for a tiny apartment since it can be collapsed and tucked in a corner
small space must-have
Mary Ann from Cape Elizabeth, ME – Verified Buyer
Good looking and fits in very small space. Sturdy when assembled but finish isn't as smooth as could be. Wood is very appealing nevertheless.
matches the description
Arleen. from Rotterdam Junction, NY – Verified Buyer
We love our butcher block table. It was not very difficult to assemble. It has worked out well for our space.
Super Table
Gloria from Warrenville, IL – Verified Buyer
This little table is perfect for my mall apartment. It's made of solid real wood and is great quality.
OUR SKU # KL1054Butcher Block Table is also sometimes listed under the following manufacturer numbers: 1622


text:
- Remember, fireworks are dangerous
- Wallace asks citizens to fight cuts
- Dispute over state payroll rolls on
- Why fight over free trade confounds partisan divide
- Still no state budget
- Crime control is not the responsibility of landlords
- Fly over to the Poplar Grove Wings and Wheels Museum benefit
- Local leaders warn of budget deadlock's impact
- SHUTDOWN: Illinois preps for the worst
- TRRT Online Edition | July 1-7
Blood drive Dec. 17 for 3-year-old cancer patient
Staff Report
Family and friends are hosting a blood drive in support of 3-year-old cancer patient Christopher Rydeberg, from 2-6 p.m., Saturday, Dec. 17, at the Verdi Club, 782 N. Madison St.
In fact, since each blood donation is separated into components, you can actually help save three lives with one donation! One in three people will need blood in their lifetime. You never know when it might be you or someone you love who will need blood.
For more information or to schedule an appointment to donate, please call (815) 968-8119.
You should be eligible to donate blood if you are at least 17 years old (16 years old with parental consent), weigh at least 110 pounds and are in general good health.
The Rock River Valley Blood Center serves as a centralized, community blood center and is the only provider of blood products and services to Edgerton Hospital and Health Services, Beloit Health System, FHN, Mercy Harvard Hospital, OSF Saint Anthony Medical Center, Rochelle Community Hospital, Rockford Health System, SwedishAmerican Health System and SwedishAmerican Medical Center-Belvidere. For more information or to make an appointment to donate blood, call toll free 877-RRVBC-99 or find RRVBC on the web at www.rrvbc.org.
From the Dec. 14-20, 2011, issue


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I used to think I wanted to be an astronaut or a father.
Nov 14, 2012 @ 1:02pm
Getting mildly turned on by the idea of Paul Ryan not having confidence anymore.
'Tis almost the season when an old person in your life gives you the gift of "texting gloves."
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text: On Mon, Nov 24, 2008 at 2:13 PM, Santiago <santi@...> wrote: > Hello > >> > Well, I think, my functions are reentrant, but I don't know yet, how I >> > can prevent fuse from creating threads (only for certain time). >> > When I change the underlaying filesystem, I'd like to block fuse, until >> > the new filesystem is mounted. >> > When I got this, I think my stuff will be thread safe. >> >> So, you have to use semaphores. You will not block FUSE, but will >> block your callbacks. > > Sorry for insisting. May be, I don't really understand the environment. > > Blocking a single callback seems not to be the problem. > I think, in parts it's already blocked, cause I do user interaction, but my > worry is, that fuse could create another thread for a quite different action. > > I would like to prevent that, so I need a semaphore, that blocks all my > callbacks at the same time. Better would be, not blocking all, but some > selected callbacks together. > Is there a way in fuse to realize that, or how can I solve that? You have to do it *in* your code. If you just want to block, not communication between the threads, you could do something like it: #include <pthread.h> pthread_mutex_t mylock = PTHREAD_MUTEX_INITIALIZER; pthread_mutex_lock(&mylock); // block the others // do something exclusively pthread_mutex_unlock(&mylock); All the conclicting code must be surrounded by the lock/unlock calls. Beware to provide an unlock to each lock call. A common error is to fail/forget to unlock a mutex and then, everything just stops waiting forever. If you have groups of callbacks where you have to block all the group when a callback belonging to this group reaches the critical code, and no matter if a member of a different group reaches the critical code, then you may declare one different mutex per group. Best Regards! -- Luis Otavio de Colla Furquim Não alimente os pingos Don't feed the tribbles http://www.furquim.org/chironfs/


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Top Definition
moldy wind is a description of farting, just like cut the cheese and other stuff
there is a moldy wind in here
i blew some moldy wind
i blew some moldy wind
by Trevor April 07, 2004
Type your email address below to get our free Urban Word of the Day every morning!
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VIEWS: 1 PAGES: 2 CATEGORY: Business POSTED ON: 3/21/2012 Public Domain
learning the region regarding revenue , promoting , money management and several various other
Tips For Building A Lifestyle Teaching Business While a lot of new starting coaches will not have the required organization knowledge to efficiently commence , run as well as deal with an enterprise there are a few percentage of those that made lots of money for some time of energy. Take into consideration coaches for example Anthony Robbins, christopher Howard, Steven covey, john Dyer, and so on. If you feel with regards to exactly why they may be consequently successful , you will find a standard twine. They are all experts as well as audio system. To efficiently build a instruction organization you must very first create the habit of smoking to create as well as discuss that you can with your particular gang of curiosity. Here are a couple tips to think about any time building your small business : 1. All living instruction training describe the importance picking a instruction area of interest. This is probably the most crucial areas where you will spend a substantial amount time to consider. The best place to start out is to look at your present knowledge , background training. Precisely what guides can you commonly go through for fun , precisely what subjects are of great interest. Picking a area of interest can be a crucial part of accomplishment. nOt just picking a area of interest is very important nevertheless picking a area of interest which has a increasing market place. Many people choose niche categories that will hardly any people are seeking. Sure there is certainly several worth because , when you're first starting out it is best to follow a successful market place , as well as afterwards branch out. 2. Build a referral network , go to organization referral get togethers , organization satisfy as well as greets , or perhaps phone your neighborhood slot provided trade for them to hook up a person with all the appropriate office to join the actual meetups 3. Keep as well as deal with a detailed customer relationship management technique. There are numerous customer relationship management technique pertaining to coaches that are ready out of the box to just plug-in the actual customers. Some of them incorporate expensive characteristics for example the ability to track advancement , supply the customer assignments ; they can even become built-in straight into your web site so as to create prospects. A person's ability to promote is very strongly in connection with a person's ability to create prospects as well as followup. The best way to figure out how to promote is to have the opportunity to master the actual craft. 4. Keep track of the accounting. Keep a very depth firewood of one's records receivable, as well as records payables. Possess contracts together with customers with regards to to pay daily activities and charges. Keeping finances tightly monitored as well as manipulated is extremely important to the accomplishment of one's organization. Think about a software like QuickBooks or perhaps likewise. 5. In case you are residing in america make sure to check out the actual small business administration place of work "small business administration " they may be any government launched firm specializing in only 1 point : enable you to be successful. They provide numerous totally free learning the region regarding revenue , promoting , money management and several various other services business coach
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