text:
|Publication number
|Publication type
|Application number
|Publication date
|Jan 4, 1994
|Filing date
|Mar 5, 1991
|Priority date
|Mar 5, 1991
|Fee status
|Publication number
|Lloyd Wolfinbarger, Scott Bottenfield
|Original Assignee
|Export Citation
|BiBTeX, EndNote, RefMan
|Patent Citations (28), Non-Patent Citations (4), Referenced by (76), Classifications (27), Legal Events (12)
|External Links: USPTO, USPTO Assignment, Espacenet
1. Field of the Invention
The invention concerns an apparatus which is suitable for permeating solid materials with various solutions and is particularly directed to processes for the demineralization of bone tissue. The invention is also directed to an apparatus for performing the processes.
2. Description of Background Information
It is known that implementation of acid demineralized bone (DB) in the form of a powder in extraskeletal sites stimulates new bone formation. Various research groups (Syftestad, 1982; Urist et al., 1967; Urist and Strates, 1970; Urist & Strates, 1971; Urist et al., 1983) have suggested that a noncollagenous protein present in demineralized bone has the ability to induce new bone formation when present within the implanted bone matrix.
Procedures presently utilized to demineralize ground bone fragments involve the use of ethanol to remove lipids and hydrochloric acid to remove the mineral component of bone.
It is also known to treat bones and bone particles to render them biocompatible so that they can be implanted in living animal and human bodies without being rejected. Included among the known methods for treating bone is the delipidization of bone using ethanol or chloroform. It is further known to demineralize bone matrix with an inorganic acid such as hydrochloric acid.
Demineralized, defatted bone can be further treated with antibacterial and antifungal agents to help prevent infection upon implantation into a living organism.
SEYEDIN et al., U.S. Pat. No. 4,434,094, describes the fragmentation of bone into small pieces and subsequent treatment by extraction with ethanol ethylacetate to remove lipids from the bone fragments. Calcium phosphate is next dissolved from the bone matrix by contacting the bone particles with dilute hydrochloric acid for one hour up to several days.
JEFFERIES, U.S. Pat. No. 4,394,370, discloses a similar method of demineralization wherein bones are first cleaned and extracted with absolute ethanol followed by anhydrous ethyl ether. The bones are then pulverized and sieved to a standard particle size. Demineralization is achieved through extraction with 0.5N hydrochloric acid for at least three hours followed by multiple washes in sterile distilled water to remove calcium and acid residues, followed by multiple sequential 60-minute washes in ethanol and anhydrous ether.
Although it is well known to defat and demineralize bone for implantation purposes, known methods of demineralizing and removing lipids have been extremely tedious, labor intensive and slow. Furthermore, an excessive amount of handling and/or exposure of the bone to non-sterile conditions has been necessary during the various phases of processing.
Delipidization and demineralization of bone matrix has been conducted largely through soaking the bone matrix in extraction solutions. Demineralization of bone involves the solubilization of hydroxyapatitic materials using fairly strong acids. As demineralization proceeds, the solubilized hydroxyapatitic materials act to buffer the acidic solution thereby raising the pH. This renders the acid less effective, and demineralization thus takes longer, or greater volumes of acid are required, because the acid solution must be completely changed to restore the pH to an effective level.
One advantage of the present invention is achieved by continually changing the acid solution during the extraction process using the apparatus(es) and processes described herein, so that the solubilized minerals are continually removed and thus solubilization of bone mineral continues with essentially similar effectiveness over time. Thus, bone processing is accomplished in less time using less total volumes of solutions.
Accordingly, it is one object of the present invention to provide an apparatus for the delipidization and demineralization of bones, bone pieces and bone particles that minimizes the necessity of exposing treated bone to non-sterile conditions. Included in this object is the provision of a treatment column which allows freezing and storing of the bone upon removal of the column from the apparatus, without requiring the bone to be removed from the treatment column.
It is another object of the present invention to provide an apparatus and method for the simultaneous delipidization and demineralization of multiple bone samples.
It is a further object of the present invention to provide an apparatus and process for delipidization and demineralization of bone samples which is less tedious and labor intensive than the methods known to the prior art. This object is to be achieved through automation of the processes involved, as well as by simultaneous processing of multiple samples.
Other objects and advantages of the present invention and advantageous features thereof will become apparent as the description proceeds herein.
Included in the description is an apparatus for processing solid matter comprising at least one column for containing the solid matter where each column has an upper end and lower end. Each lower end has an inlet valve attached and each upper end has an outlet valve attached. Each column is adapted to contain solid matter during the extent of processing. The apparatus further includes a driving mechanism for conducting flow of solutions through each column, which is preferably at least one peristaltic pump, for processing the solids therein; column clamps for supporting each column in a vertical position during processing; and solution reservoirs for holding the solutions which are to be conducted through the columns. Each column is adapted to be removed from its supports after processing and each inlet and outlet valve is adapted to be closed to allow for sterile storage of the solid matter located in each column.
Preferred solid materials to be processed in the apparatus include particulate bone and strips of cortical bone. The entire apparatus can be contained within a refrigeration system which is capable of maintaining the apparatus at a processing temperature range of 0� to 38� C.
The invention further includes detecting devices for detecting solutions which exit the outlet side of the column, which are referred to as eluent. The detecting device(s) may detect pH, calcium ion concentration or conductivity. Included among the detecting devices are calcium specific electrodes for detecting calcium ions Detecting devices may also include flow rate detectors which connect each solution reservoir with a solution switching mechanism and the driving mechanism. Flow rate detectors may include optical drop counter devices which connect each solution reservoir with a flow switching device which connects to the driving mechanism.
The present invention further may include a computer controller for automated control of flow through the apparatus; on the other hand, the apparatus may be manually controlled. The detecting devices may provide feedback to the computer control means to enable the computer to control flow and switching of solutions.
Flow control and switching of solutions may be pneumatically driven or electrically driven by solenoid controllers.
The outlet side of each column may include a mechanism for preventing backflow of contaminants, which is preferably an optical drop counter device.
The invention is also directed to a process for extracting undesirable constituents of a solid material, which comprises loading a solid material into at least one column, where each column has an inlet valve and an outlet valve; flowing a solution through each column; detecting a characteristic of a solute which is present in the solution as the solution exits each column; terminating flow of the solution once a desired characteristic of the solute is detected and thereafter flowing a different solution through each column.
The process further encompasses detecting a characteristic of a second solute which is present in the subsequent solution as the solution exits each column; and terminating flow of the solution once a desired characteristic of the second solute is measured.
Furthermore, the process encompasses flowing additional solution through each column and detecting a characteristic of each solute which is present in each solution as each solution exits each column.
The solutions employed in the process include lipophilic solvents, sterile deionized water, buffers and acids.
The lipophilic solvents may include ethanol, chloroform or other lipophilic solvents known in the art. The acids employed include dilute hydrochloric acid and other organic or inorganic acids known and used in the art for demineralization of bone.
Detection methods for the lipophilic solvents include detecting the absorbance of the solute dissolved in the lipophilic solution, and dripping a sample of the lipophilic solution into deionized water to determine whether a precipitate forms. Detection methods for the acidic solvents include measuring the pH of the acidic solution, detecting the presence of calcium ions in the acidic solution by measuring the calcium ion concentration of the solution, and measuring conductivity.
The flow control for the processes may be accomplished by at least one pneumatic control valve or at least one electric solenoid controlled valve.
Each flow control valve may be computer controlled or manually controlled. Any combination of or all of the detecting devices may provide feedback to a computer controller for providing information useful to the computer to control flow or to decide when to switch solutions.
The columns of the apparatus may be readily sealed and removed from the apparatus, with the processed solid material still within the columns, for sterile storage of the processed solid material.
Also included in the invention, is a fine mesh bag which may be preloaded with solid material and then loaded into the column. The fine mesh bag is preferably used for dental bone and other small solid particles in the range of 250μ to 700μ. The bag is preferably formed of polyester but may be made of other materials which are resistent to and nonreactive with the solutions that it will interface with during processing. The fine mesh bag has a porosity which is small enough to retain the smallest solid particles with which it will be loaded, yet large enough to permit the escape of gases which are formed during processing.
The present invention is further explained in the description which follows with reference to the drawings, illustrating, by way of non-limiting examples, various embodiments of the invention, with like reference numerals representing similar parts throughout the several views, and wherein:
FIG. 1 is a plan view of a complete bone demineralization apparatus according to the present invention;
FIG. 2 is a schematic diagram of a control system for an apparatus using a plurality of demineralizing columns and solutions adapted to be conducted through the columns;
FIG. 3 is a schematic diagram of a control system for an apparatus using a single demineralizing column, as basically shown in FIG. 1, and solutions adapted to be conducted through the column;
FIG. 4 is an embodiment of the top portion of a column shown in FIG. 1 which is adapted for processing, e.g., dental bone;
FIG. 5 is a bottom portion of a column adapted to engage the top portion shown in FIG. 4;
FIG. 6 shows a longitudinal sectional view of the bottom component of FIG. 5, together with an inlet valve;
FIG. 7 is a cutaway longitudinal sectional view of the column components of FIGS. 4 and 5 in assembled form;
FIG. 8 is another longitudinal sectional view of the processing column of FIGS. 4 and 5 which depicts uniform flow;
FIG. 9 is a view of a column similar to that of FIG. 4, with a top filter which is used to retain solid matter within the processing column;
FIG. 10 is a sectional view of another embodiment of a column similar to that of FIG. 9, and which includes a compacting plunger;
FIG. 11 is a plan view of a fine mesh bag which is used for processing, e.g., dental bone;
FIG. 12 is an exploded plan view of the fine mesh bag of FIG. 11;
FIG. 13 is a plan view which indicates the loading of a fine mesh bag with the aid of a powder funnel;
FIG. 14 is a plan view of a fine mesh bag with its filling spout in the closed position;
FIG. 15 shows the connection of an inlet tubing with the inlet port of a processing column; and
FIG. 16 shows the filling of an alternate embodiment of a column of FIG. 1, which does not require a fine mesh bag.
Referring now to the drawings in detail, FIG. 1 shows a complete apparatus for the delipidization and demineralization of bone samples. Generally cylindrical column 1 is removably secured in a vertical position by column clamp supports 33. Columns clamp supports may, for example, be formed by rubber coating spring steel of the required dimensions. Containers for holding solutions A, B and C each have sterile in-line filters 35a attached upstream of their respective inputs to prevent bacterial and other contamination of the sterile solutions contained therein. The invention is not limited to three containers (e.g., reservoirs) for holding solutions as shown by the embodiment in FIG. 1, but may use more or less than three reservoirs, depending on the number of different solutions that are to be used for the particular process in question. For example, one may wish to further treat the demineralized bone subsequent to defatting and demineralizing with antibacterial solutions, antifungal solutions, vitamins, bone morphogenetic protein or other osteogenic factors, collagens or other proteins, etc.
Furthermore, the apparatus is not limited to use only for demineralization of bone. It can be used advantageously in many other instances where it is desired to permeate a solid/particulate material with one or more solutions. It is particularly useful when it is desired to maintain the solid/particulate material under sterile conditions during processing. Other possible uses of the apparatus include, but are not limited to: crosslinking of collagen-containing implant materials; treatment of natural blood vessels to render them nonantigenic; the treatment of soft tissue implants with, for example, heparin to render them non-thrombogenic. Another set of in-line filters 35b, similar to filters 35a, is attached to the outflow side of reservoirs A, B and C, upstream of the respective outflow valve for each line.
Optionally, flow rate monitors 37, which may for example be optical drop counters, may be connected between the outflow side of reservoirs A, B and C and in-line filters 35b, for providing feedback to controller 38 for flow control. In this embodiment, a 3 to 1 manifold includes switching means for controlling whether solution A, B or C is to pass through to peristaltic pump 36.
The peristaltic pump 36 which is illustrated in FIG. 1 is not necessarily required to drive the flow through the apparatus. Other pumps and driving mechanisms known in the art can be used. Yet another in-line filter 35c is attached in line with the outflow side of pump means 36. From the outflow side of the in-line filter the tubing is next attached to inlet valve apparatus 12 which is attached to the inlet part of the column at the bottom end 7 of the column. Bed support 34 may be placed on top of bottom end 7. Optionally, a manifold (i.e. 1:3, 1:4 or some other ratio) can be connected to the outflow side of pump 36 to enable parallel processing of multiple columns, as shown schematically in FIG. 2.
The tubing that is attached to the outlet valve 2 of the column leads to a waste receptacle where the eluent is disposed of. A flow rate monitor 40 may optionally be connected in line between each column and the waste receptacle to prevent backflow of waste eluent into each column. Furthermore, a monitoring device 39 may be attached in-line to the outflow side of each column, before the waste receptacle and before, the optional flow rate monitor 40. Such monitoring device may include, e.g., photometers, pH monitors, calcium ion specific electrodes and conductivity probes. Monitoring (detecting) device 39 provides feedback to controller 38 (e.g., a CPU) to enable controller 38 to decide when to change the flow from one solution reservoir to the next.
The entire apparatus described above is preferably contained within a refrigeration cabinet 31. A conventional refrigeration system 32 is used which is capable of maintaining the apparatus in the range of 0� to 38� C. All apparatus components are adapted to be removed from the refrigeration cabinet 31. The apparatus can be controlled manually or automatically. As stated above, the flow meters and other monitoring devices 37, 39 and 40 may provide feedback to a computer 38 for automated control of solution flow.
FIG. 2 is a schematic diagram of an embodiment of the present invention for processing multiple samples simultaneously in a plurality of columns. In this embodiment, four solution reservoirs (A, B, C and D) are connected to flow control mechanism 44. Flow control mechanism 44 (e.g., a plurality of flow control valves) may be either pneumatically driven or driven by electric solenoid and may be manually controlled or controlled by computer 38. If the computer option is chosen, the computer should be programmed, with the option to be reprogrammed, to control the time intervals of the control mechanism 44. The peristaltic pump(s) can be set to deliver a flow rate that is independent of the controller input. The flow is passed through e.g., a 4:1 manifold 43, and then through e.g., a 1:4 manifold 41 in this embodiment to connect to two peristaltic pumps 36. Each peristaltic pump is connected to two columns 1 so that four samples may be processed in parallel. Eluent exits the columns at 42, in the fashion described above with respect to the column of FIG. 1.
Flow control mechanism 44 shown in the schematic drawing of FIG. 2, includes flow control monitors which provide feedback to computer controller 38, so that the controller can determine whether to maintain, increase or decrease the detected flow rate(s). Feedback to unit 38 is also provided by monitoring device 39 (not numbered) at the eluent side of the columns 1, so that the controller can determine when each stage of processing is complete. The controller, upon each such determination, shuts off flow from the solution reservoir that is then supplying solution via flow control mechanism 39, and initiates flow from the next solution reservoir in the processing sequence.
Optionally, flow control valves (not shown) may also be located between 1:4 manifold 41 and peristaltic pumps 36, and these valves may be optionally controlled by computer controller 38 or manually controlled. Flow control valves, when positioned into this location, allow differential processing of parallel columns 1. For example, one column may contain smaller particles than the other three and require less demineralization time. In such a case, controller 38 would sense, through feedback from a pH or calcium ion monitor 39, that demineralization is complete in one column before it is complete in the other three columns. The controller would then close the valve near the peristaltic pump which leads to the completely demineralized sample, while hydrochloric acid would continue to be pumped through the remaining three columns. The optional flow control valves would, of course, also permit one or more columns to be completely shut down when less than four samples are to be processed.
FIG. 3 is a schematic diagram of an embodiment of the present invention for processing one sample at a time in column 1, as essentially illustrated in FIG. 1. Three solution reservoirs (A, B, and C) are shown, but more or less reservoirs may be supplied as discussed earlier. Flow control mechanism 44 is similar to that shown in FIG. 2; but in this instance, only three lines are controlled in and out of the mechanism. Flow continues to a 1:3, manifold 45 and a single peristaltic pump 36. An optional flow control valve is not necessary at this location for this embodiment, since flow is either existent or non-existent to the single column in this embodiment; and because this single column can be controlled at device 44.
Similarly to FIG. 2, computer controller 38 receives feedback from the monitor(s) at the eluent side of column 1 and from the flow rate monitor(s) at (or upstream of) the location of control mechanism 44, for control of solution source switching and flow rate, respectively. As in the embodiment of FIG. 2, controller 38 comprises, e.g., a CPU capable of controlling flow in response to the detected flow rate(s) and eluent.
FIG. 4 shows a preferred embodiment of the top component of a column 1 useful in processing, e.g., "dental bone" (i.e. bone particles ranging in size from 250μ to 700μ). This device is not limited, however, to use for the demineralization of dental bone, but also in processing finely ground bone particles, and may also be used in any production process requiring the retention of some desired size range within a permeable bag while solvent solutions are passed over and through the particles to affect a washing or extraction, or deposition of, solute onto the particles.
The top end of column 1 includes a threaded outlet port 4 to which an outlet valve 2 is connected via screw threads. The top end also contains a levelling device 3 which assists in fixing the column in a vertical position to help ensure uniform flow of solvent during processing. Levelling device 3 can, for example, be a well-known bubble type level. When the column is positioned vertically, the bubble in the level centers in the fluid contained in the level.
At the bottom end of column ? , is located a flange 6 which accommodates an "O" ring (not shown) for securing the connection to be made between the top and bottom components of the column. Protruding through flanges 6 are threaded connecting bolts 5, which are preferably formed from teflon or polypropylene.
Preferred materials for constructing the column are glass or polycarbonate, but other construction materials known in the art which are inert to the solutions to be passed through the column may also be used.
Preferred dimensions for column when used for processing dental bone, are 7.5 cm for the height of the column and 21.5 cm for the diameter of the column. These dimensions are appropriate for the quantity of bone matrix expected to be obtained from a tissue donor. To accommodate more or less quantity of bone material, the height of the column should be adjusted while leaving the diameter unchanged. The height of the column can be increased to optimally accommodate larger size particles of bone material, but this is not absolutely essential. Especially for smaller bone particles, selection of the appropriate column geometry appears to be important in permitting the escape of CO2 gas formed during processing. The escape of CO2 gas helps prevent the bone particles from compacting since the particles swell as demineralization proceeds, and ensure uniform flow of solutions through and over the bone particles. With larger bone particles/pieces, the dimensions of the column appear to be of minimal importance, requiring only that an attempt be made to reduce the volume of solutions inside the column not occupied by the bone.
FIG. 5 shows a bottom component 7 which is designed to connect with the bottom end of column 1, via bolt holes 9. A threaded inlet port 8 located at the center of the member 7 accommodates an inlet valve mechanism and conducts inflow of solvents into the assembled column.
FIG. 6 is a sectional view depicting the engagement of an inlet valve mechanism 12 with inlet port 8 via threads 13. A preferred valve mechanism is a 1/4" or 1/8" MULTIFIT � valve, constructed of teflon, but any two-way valve mechanism which is both resistant to the solvents to be passed through it and capable of sealing inlet port 8 in a sterile fashion may be used. FIG. 6 also shows a fine mesh screen 11 which is designed to fit over the center portion 10 of annular bottom component 7, and is designed to prevent the smallest solid particles from falling down through the apparatus while facilitating a uniform inflow of solvent. A preferred fine mesh screen for processing dental dust is formed from polyester cloth having about a 225μ pore size. Fine mesh screen 11 is sealed in position by an "O" ring 14, which surrounds downwardly depending edges of the screen. It can thus be relatively easily removed, if it deteriorates or becomes clogged, and can be cleaned and/or replaced by first removing the flexible O-ring.
FIG. 7 is a sectional view of a preferred embodiment of the bottom member 7, and the lower end of column 1, in assembled form. Connecting bolts 5 pass through bolt holes 9. Wing nuts 16 are torqued onto the connecting bolts to enable "O" ring 15 to provide a seal between the top and bottom components.
The center portion of the bottom component 7 is tapered, as shown by the sectional view in FIG. 8, to effect a more uniform flow of solvent 18 through the solid material located within the column. FIG. 8 also shows an "O" ring 17 which provides a sealed connection between the inlet valve mechanism and the inlet tubing.
FIG. 9 illustrates an assembled column, as used in the system of FIG. 1, which has been loaded with bone particles 20. Top filter 19 prevents the outflow of particles and is preferably constructed of glass wool wrapped in cheesecloth. However, any screening material which is resistant to the solvents used in processing and inert to the reactions involved in the column, with a porosity smaller than the smallest particles in the column yet large enough to provide free outflow of solvent, may be used.
It is important that the bone particles occupy 80 to 90% of the column volume so that an optimum spacing between the particles and solvent flow therebetween is maintained. When a "normal" quantity of bone matrix (i.e., 300-400 gm) is to be processed, the column of FIGS. 4-8 is configured so that the sample occupies 80 to 90% of the column volume. As stated earlier, the column height of the preferred embodiment can be altered when the volume of bone to be processed is known prior to use. However, when a bone sample volume is not known in sufficient time to calculate and construct a column with a desirable height, and, therefore, when a column of proper dimensions cannot be predesigned, a plunger 21 may be positioned at the top of the column and slid into the column 1, as shown in FIG. 10 to reduce the effective volume of the column so that the bone particles occupy 80 to 90% of the effective volume of the column. The plunger 21 has an annular post which runs centrally through its length to allow solution to flow through to the outlet of the column.
FIG. 11 shows a fine mesh bag 22 which is preferably used as a container for the processing of dental bone or other bone. The fine mesh bag facilitates retention of the ground bone particles during processing. The fine mesh bag is preferably formed of polyester cloth which is resistant to absolute ethanol and up to 1.0N hydrochloric acid. The bag is formed of three panels as shown in FIG. 12. Circular discs 24 and 25 each have a diameter that is equal to the outside diameter of top component 1. Narrow strip 26 has a width which is approximately 1/2" less than the height of column 1. The bag has a porosity of around 225μ and is thus of a small enough porosity to retain the smallest bone particles in the dental dust, which have a size of about 250μ, yet has a large enough porosity to permit formed gas (i.e., CO2) to escape.
Loading of the fine mesh bag 22 is facilitated by a commercially available powder funnel 27 as shown in FIG. 13. Bone particles are loaded through bag filling spout 23. FIG. 14 shows the fine mesh bag fully loaded with bone particles and ready for insertion into the column, with the filling spout 23 having been folded around the outside of the bag. No closure devices are required to close the filling spout, since the snug fit of the filled bag against the inner walls of the column prevents the spout from opening. The filling spout is shown in the open position by phantom lines 23'.
The fine mesh bag at this stage may be placed into column 1, with bottom component 7 then being attached and bolted into place.
Further advantages obtained through the use of the fine mesh bag during processing are: easy addition and removal of the bag from the column; facilitation of a uniform flow of solvent through the bone particles in the bag; expansion and movement of the mass of bone particles in the bag as CO2 gas is formed. The formation of CO2 gas during demineralization effects a "stirring" of the confined bone particles which results in a more uniform demineralization of the particles. If the bone matrix initially takes up greater than 80-90% of the column volume, there is a danger of compaction of the bone particles when CO2 gas begins to form, thereby restricting demineralization. The fine mesh bag further permits expansion of the bone mass since the bag material is deformable and resilient. The bone particles deform the bag upon expansion and when the bag resiliently returns to its original configuration, a stirring effect occurs. It is important that the outside diameter of the fine mesh bag fits easily and snugly into the inside diameter of the top component of the column device. A snug fit restricts the flow of solvent around the bag, thereby forcing the solvent flow through the bag so that a uniform flow of solvent interfaces with the bone particles therein, and also maintains the spout in a closed position.
FIG. 15 is a sectional view showing the connection of inlet tubing 28 to inlet valve means 12. Threaded cap 29 engages the threads on inlet valve means 12 to enable "O" ring 17 to sterilely seal the connection.
FIG. 16 shows an alternate method of loading a column which is more often used for bone particles of larger dimension than dental bone. Funnel 30 facilitates direct loading of bone particles into column 1, without the use of a fine mesh bag positioned within the column. Columns may also be loaded directly with, e.g., cortical strips of bone for demineralization.
Once the column is filled and the bottom component is connected with the column, the column is assembled into the apparatus as shown in FIG. 1.
Advantageously, once the bone sample has been loaded into column 1 and connected in line with the apparatus of the present invention, it is sterilely sealed and there is no requirement to ever break the sterile seal until such time as the sample is to be freeze-dried and dispensed. More specifically, the column is connected to the inflow and outflow lines of the apparatus and inflow and respective outflow valves 12 and 2 are opened to permit solutions to be selectively flowed through column 1. Both inflow and eluent (outflow) sides of the column ensure sterility of the column with the various in-line filters. The flow control detector 39 and controller 38, as described earlier, permit the switching of different solution flows through the column without ever having to disconnect and reconnect any tubing The advantage of this is that the entire system remains sealed and sterile.
Upon completion of flow of all processing solutions through columns 1, inflow and outflow valves 12 and 2 are closed to sterilely seal the inner contents of each column from the rest of the apparatus. Each column can then be removed from its clamp supports 33 and stored for future use. Optionally, adapters (which can have, e.g., flat support surfaces) may be threaded onto the inflow end of inflow valve 12 and the outflow end of outflow valve 2, to enable secure storage of columns on a vertical (or other) orientation. Thus, there is no requirement to expose the treated sample to a non-sterile environment and the sample can be frozen in its sterile condition within column 1.
Further details of the process of the invention are presented in the examples that follow:
Properly sized ground bone (250μ to 700μ) is placed into the fine mesh bag, as described above, and placed into the column. The column is closed and placed into a clamping device such that it is level.
Utilizing a peristaltic pump, solvents are pumped into the column through the inlet port and outwardly from the outlet port. The valves placed at each of these ports are designed to permit sealing of the system prior to and after processing to ensure that the bone materials will remain sterile. Bone particle processing proceeds as described:
1) Ethanol (95% Ethanol, 5% water [deionized]; sterile) is pumped rapidly into the column until full. At this point, the bone particles may be left to "soak" in the ethanol for a prescribed time interval, with changes in ethanolic solution (i.e., adding fresh solution to replace the original volume of ethanol), until ethanol extractable lipids are extracted. Alternatively, ethanolic solution may be continuously pumped through the bone, at a flow rate of approximately one (1) ml per minute. At this flow rate, approximately 84 mg of ethanol soluble lipid should be extractable per 10 gms of bone matrix. This value will vary from donor tissue to donor tissue. The volume of ethanol required should approximate 90.0�5.49 mls/10 gms of bone, although this value may also vary with donor tissues. Elution of ethanol soluble lipids may be monitored by measuring absorbance of eluent solution at 210 nm. As an alternative, less sensitive assay eluent may be dropped dropwise into deionized water in a small test tube. If lipids are present in the eluent ethanol, a cloudy precipitate will appear in the upper part of the aqueous volume as the dissolved lipids come out of solution
Sterile deionized water is pumped into the column to displace the ethanolic solution. Alternatively, the ethanolic solution may be replaced with acid solution such that demineralization commences immediately. Problems to watch for at this point include, but are not limited to, the potential for formation of lipid micelles if ethanol soluble lipids are still present to an appreciable extent in the ethanolic solution. Such lipid micelles may absorb onto the bone particles. The effects of such absorption on activity of bone product (i.e., osteoinduction) are unknown at present. Demineralization of bone particles is accomplished by pumping 0.5N HCl through the bone particles at a flow rate of approximately one (1) ml per minute. Data developed by the inventor indicates that minerals (assayed as CaCl2) should be extracted at approximately 0.042 gms/ml acid and that extraction should be "complete" (i.e. less than 5% by wt. calcium) with use of approximately 160 mls acid per 10 gms of bone. The values are projections based on the use of larger bone particles in a less favorable column geometry and thus, although values will vary from donor tissue to donor tissue, it is expected that with the smaller bone particles in dental bone, minerals will be extracted more efficiently utilizing less volume of acid. "Completion" of demineralization may be monitored utilizing a calcium-specific electrode, measuring calcium ion concentrations in the eluent, or by monitoring pH of the eluent. A potential problem during processing is the formation of gas bubbles as the acid contacts the bone particles. It is suggested that this gas is CO2 evolving as the acid reacts with calcium carbonate in the bone. If acid is pumped too rapidly, gas formation can be excessive and should the outlet port become clogged, pressure can build up in the device and cause leaks around the "O" ring between the column and its member. The formation of gas can be beneficial, in that it "stirs" the bone particles as it rises upwards through the bone particles, escaping through the outlet port.
______________________________________Column Information:______________________________________Height 30 cmDiameter 1.4 cmVolume 52 mlWeight of bone 14.532 gHeight of bone 13.5 cmat commencement of experimentHeight of Cheesecloth 8 cmat commencement of experiment______________________________________
A column of bone particles were filled with absolute ethanol and allowed to soak for 16 hours. A volume of 143.8 ml. of ethanol was then flowed through the column at a flow rate of 1.2 ml./minute. The delipidization phase was then terminated and a total volume of 200 ml., of sterile distilled water was flowed through the column at a rate of 6.8 ml./minute.
The demineralization phase began by running 228 ml. of 0.5N HCl through the column at a rate of 0.95 ml/minute. The eluent pH at the end of this run was 0.26. Another volume of 200 ml of sterile distilled water was then run through the column at a rate of 0.19 ml/minute.
Five pieces of bone, which were still somewhat rigid, were removed from the column and lyophilized. The weight of the bone before lyophilization was 7.336 g. The weight of the bone after lyophilization was 5.793 g.
The remaining pieces of bone in the column received 1800 ml more of 0.5N HCl at a flow rate of 0.43 ml/minute. The eluent pH at the end of this run was 0.28. A volume of 200 ml of sterile distilled water was then run through the column at a flow rate of 6.8 ml/minute.
The remaining pieces of bone were removed. These pieces, being pliable, were lyophilized. The weight before lyophilization was 5.266 g, and after lyophilization the weight of these bone samples was 1.922 g.
Cortico/Cancelleous bone fragments were ground to a heterogeneous size. Approximately 90 gms of bone matrix was packed into a column (2.5 cm I.D. by 60 cm in height). The height of the bone matrix was approximately 40 cm. A small "ball" of glass wool wrapped in cheese cloth was placed on top of the bone matrix and a column plunger was placed into the column and run down to slightly compress the ball of glass wool/cheesecloth. The column was sealed and absolute ethanol pumped through the matrix at a flow rate of 3.7 ml/minute. Lipid in the eluent was monitored by dropping ethanolic eluent into water and observing for a cloudy precipitate. In this instance a more sensitive form of this assay was used, collecting 6.8 ml fractions and adding 6.8 ml of water, watching for a cloudy precipitate as an indication of the presence of lipid in the eluent. Complete removal of ethanolic extractable lipids required 2,477 ml (and included an overnight "soak" of the bone matrix in ethanol). A total of 390 mls of distilled water was used, at a pump rate of 3.7 mls/minute, to remove the ethanol (390 mls represents the approximate volume of the column, i.e. one column volume of water) was used to elute the ethanol. With a flow rate of 3.7 ml/minute, this "washing" required approximately 100 minutes (1 hr., 40 min). Then, the solvent is switched to 0.5N HCl, at a flow rate of 3.7 ml/minute, and the acid is pumped through the bone to affect demineralization. By monitoring pH, demineralization was essentially completed by 300 ml (Note: calcium ion concentration in the eluent acid always paralleled and followed the pH profile. At a flow rate of 3.7 ml/minute, demineralization thus required approximately 82 minutes. Buffer was not run over the bone, as it was preferred to run distilled water (1,580 ml). The pH was returned to approximately 6.2. The bone matrix was then freeze-dried for 2 days and weighed. The weight of the freeze-dried bone was 40.33 gms.
The experimental protocol for the demineralization process of Example 3 is as follows:
1) Grind bone matrix as per standard protocol for producing cortico/cancellous bone matrix. Expect bone matrix to approximate 90 to 100 gms as a "donor lot". This bone mass may be exceeded by 10 to 20% and still utilize the column indicated below. All procedures were conducted at room temperature.
2) Screw the bottom plate onto a 60 cm�2.5 cm (I.D.) glass column and pour the bone matrix into the column. Shake gently to "compact" the bone matrix. Do not compact tightly.
3) Place a small ball of glass-wool wrapped in cheese cloth on top of the bone matrix and insert the column plunger until it slightly compacts the ball. Do not compress the bone matrix. Seal the column endplates and open valves after attaching tubing.
4) Place the column, with bone matrix, into the column holding device, bottom plate down and attach the tubing to the solvent distribution apparatus. Place outlet tubing into waste container. All solvents will be run bottom to top of bone matrix.
5) Run 95% ethanol into the column, until it begins to flow from the outlet line (approximately 100 to 200 mls, depending on bone matrix and corresponds to a "column volume" as used below). Let stand at room temperature for 30 minutes.
6) Run an additional volume of 95% ethanol into the column and let stand for 30 minutes. Repeat this to a minimum of one additional time (3 to 4 times total), i.e. a total volume of ethanol of 500 ml for every 100 gms of bone matrix and a total extraction time of approximately 2 hrs. Note: To monitor for lipid extraction, add approximately 5 ml of ethanol eluent into a test tube and add an equal volume of water. A cloudy precipitate indicates that lipids are still present in the eluent ethanol. Lipids will most probably be present.
7) Run 0.5N HCl onto the column at a flow rate of approximately 4 ml/minute and monitor the pH of the eluent, beginning after approximately 200 ml of acid. When pH has fallen to approximately pH 1.0 to 2.0, run acid for an additional 100 ml of acid.
8) Run 3 column volumes (a column volume equals that volume of ethanol that is required to fill the column initially) of distilled water over the bone matrix. This flow rate may be that which is maximally possible based on the tubing diameter utilized.
9) Run 3 column volumes of 0.02M potassium phosphate buffer (pH 7.4) onto the column, at maximal flow rate. Check the pH of eluent solution near the end of the buffer run. It should be approximately 7.0 to 7.4).
10) Run 6 column volumes of distilled/deionized water onto the column, maximum flow rate possible. If you wish to check pH near the end of this run, add 2 ml of 1M KCl/100 ml of solution and check pH using a calibrated pH meter. It should be approximately pH 6.
11) Close column valves (both ends) and remove column from the column holding device and invert the column. Disconnect tubing from solution distributing device and open valve on column bottom and then valve on column top (this means original top and bottom, not present top and bottom) and allow solution to drain from the column into a waste container. Care should be taken at this point not to introduce microbial contamination, and optionally an in-line air filter may be added to the "air inlet" line. To effect this draining, the column may be reclamped in the column holding device "up side down" and left to drain.
12) Remove bone matrix from column after removing end plates and/or plunger. Dry.
Thus, a preferred process includes the loading of bone particles or pieces into at least one column, either directly or by first loading the bone into a fine mesh bag and then placing the fine mesh bag into the column. Each bottom plate is then sealed and secured to each column, and each column(s) is secured in a vertical position with the column clamps.
Inlet and outlet tubing is connected to the input and output valves respectively. The flow of solutions is then commenced through the column(s). Ethanol is generally the first solution that is passed through the column to delipidize the bone. Sterile distilled and/or deionized water may optionally be run through the column after the ethanol run, to wash the bone, or an acidic solution, preferably HCl may be run immediately following the ethanol run to demineralize the bone. Optional solutions may be passed through the column(s) after the demineralization run is complete, to include, sterile distilled and/or deionized water; solutions of vitamins, antibacterial agents, antifungal agents, bone morphogenetic protein and the like.
Solution flow begins at the solution reservoirs and proceeds through optional flow meters, through in-line filters, then through a flow control/switching mechanism which may be manually or computer controlled. The flow control/switching mechanism may be pneumatically driven or driven by electric solenoid. Flow then proceeds to at least one pumping mechanism which provides the driving force for the flow throughout the entire apparatus. The pumping mechanism pumps the solution through an in-line filter through each inlet port and into each column. The solution interfaces the solid bone particles/pieces as it flows through the column(s), exits the outlet port(s), passes through optional detecting devices, optional flow rate monitor and into a waste receptacle.
The flow rate monitors and detecting mechanisms provide feedback to the computer controller. The computer controller uses the feedback to maintain desired rates of flow of solutions, and to determine when each particular phase of processing is complete Upon completion of a phase of processing, the computer signals the flow control/switching mechanism to shut off flow from the current solution reservoir and to open flow from the next successive solution reservoir. The computer controller is programmed to continually monitor solution flow and to switch solution sources at the proper times until processing is complete.
Upon completion of the delipidization/demineralization process, including any washing steps or other treatments mentioned earlier, the switching mechanism is directed to shut off all flow of solutions through the column(s). The inlet and outlet valves are then closed on the columns, and after draining the column(s) of any remnant solution, the bone samples may be directly stored in the column(s) in a sterile condition.
Although the invention has been described with reference to particular means, materials and embodiments, it is to be understood that the invention is not limited to the particulars disclosed and extends to all equivalents within the scope of the claims.
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|U.S. Classification
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|International Classification
|Cooperative Classification
|European Classification
|Mar 5, 1991
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text:
The Maumelle Monitor reported that I spoke in favor of an indoor pool at the last City Council meeting. Evidently I did not make myself clear, so let me be clear now. I am NOT in favor of Maumelle taking on the up-front and continuing cost of an indoor pool at this time.
I did say that I understand that a therapy pool would benefit seniors and that competitive swimmers would like an indoor pool for year-round training. However, the real question isn't whether or not folks would like an indoor pool. Clearly, many would. The real question before the Council is "Can Maumelle afford one?" I think not. It has been asserted that the various charges for activities at an indoor pool would definitely cover its operational costs. But that assertion has not been accompanied by credible proof. Other Arkansas towns have had to raise taxes to cover the costs of an indoor pool. Some communities that have built a pool have had to drain and close it due to a lack of funding.
Just like families, cities have to budget for known needs, and they have to have enough financial reserves to cover unanticipated expenses. Maumelle also needs to have enough money to pursue important steps in our Strategic Plan for the future. To do otherwise would be to waste the investment already made in that project and to short-change the better future that Maumelle could have.
Would I like an indoor pool? Sure. I'd also like a personal chef. But the sad fact is that neither the City nor I can finance those "wants."
Terry Jones

subset: commoncrawl


text:
Atheists and Agnostics Gather Here... (Part 2)
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Stazon Splitcoast
Join Date: Dec 2006
Location: my heart remains in France
Posts: 21,989
Originally Posted by
Duh, guess that would help, LOL:
Losing Your Religion: Analytic Thinking Can Undermine Belief: Scientific American
Oh, and here's another article that gives a few more details of the original journal article:
If You Answer This Math Problem Correctly, You May Be an Atheist
Did you scroll down to read the comments??? Hilarious!!! I love the comment that says "math is stupid, just like religion". (I like math)
I got both answers wrong, despite liking math. As the (current) first comment says, "I have failed as an atheist". LOL
Anyway, I was once a believer and now am not, not because of analytical thought or the internet. Sometimes, I think I still DO believe that God exists, because of the rage that simply arises in me against God (I think if I TRULY did not believe that there was a god, I wouldn't have this rage - I mean, I NEVER rage against Santa Claus). It's just that while I believe that (the Protestant) God exists in those moments of rage, I disagree with Him vehemently and viscerally and think it is unconscionable that He allowed SO much ugliness into the Bible that allows some (okay, in my personal opinion, it is a LOT) of His followers to behave in such extremely awful, cruel, and mean ways. I guess the turning point was decades ago when a (straight) couple asked me WHY it was so wrong for gays to marry. I gave the pat religious answer, but hated myself for it. I felt horror at what I had said. I guess that was more the final straw than anything. Years and years of having given pat religious answers and feeling like a idiot parroting hypocrite and the resulting self-hatred and self-disgust crashed down on me and I finally "came out" as a "non-believer".
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text:
June Summers ft. Sean Kingston - Grinding
While June Summers recently stepped into the Booth with compilation mixtape Hookman for Hire, it's been a solid year-and-a-half since he last brought us a solo feature. As Steve Perry would say, though, "Do-o-n't stop, belie-e-evin!" Far from having fallen off, the NYC repper is, was and ever shall be "Grinding on and on and on and on…" – and he's back with a Booth-exclusive world premiere to prove it. If you didn't catch my hint, this new single is an homage to Journey's Don't Stop Believin', with (who else?) "J.R." Rotem flipping the '81 arena-rock hit's iconic keyboard chords into a triumphant backdrop for lyrical reflections on June Summers' epic hustle. In bars that are self-congratulatory but not self-satisfied, the singer informs listeners that his indefatigable work ethic is an inborn trait: "[Can't] take the hustle out of me, I got it biologically, my father handed it down to me, so I guess I got it honestly" Topped off with the anthemic hook vocals of Caribbean-pop heavyweight Sean Kingston, this cut could hardly be a grander re-entrance. If you're left wanting more, keep it locked for release details on Summers' forthcoming Almost Famous LP.


text:
Advice for breeders
Introduction to Breeding
Please consult a vet before you consider breeding a bitch, the information provided is no way a substitute for veterinary care and advice. Breeding is not something to be taking lightly, it is a big responsibility and something that should be taken very seriously, if you would like to become a breeder it is strongly advised that you seek advise from you bitches breeder. There is not much money to be made in breeding so never breed to try and make money – breeding should only be to improve the breed.
Why do you want to breed?
If you want to breed because you want to make money, then please reconsider breeding. Breeding is expensive and you are no likely to make any money from it. You must consider vet bills, food, equipment and advertising.
Do you have the room?
Puppies may be cute but they take up a lot of space and can be very messy! You will need a large run and whelping box and large run large enough for up to 12 puppies. Bedding will need to be washed once or twice a day and a secure outside area is important so puppies can run around and play.
Do you have the time?
Your bitch will need you there in the days leading up to her labour and you will need to be present and on hand for the whelping which can take 24 hours!
It must also be considered that your bitch needs a special diet and frequent walks, later on when the puppies are older you will also need to feed, walk and take care of them all. Taking care of up to 12 puppies for 7 weeks requires a lot of time and dedication.
Do you have the knowledge?
Do you know what to do should anything go wrong? If the answer is no then you need to research and educate yourself before breeding. Breeding is a serious responsibility and not something to be taken lightly – the more knowledge you have the better change you bitch and puppies have of remaining fit and healthy. The puppies potential new owners will also look to you as a source of information and its your responsibility to be at hand to answer any questions they may have.
Are you aware of your legal responsibilities?
The Breeding and Sale of Dogs Act 1999 states that if you are going to embark on a career breeding dogs you must obtain a license is the bitch gives birth of 5 or more liters in any 12 month period.
During an inspection you must adhere to the following areas:
Record keeping including
- identifying marks (tattoo/microchip, kennel club registration)
- disease and infection control
- matings
- whelpings
- oestrus
- numbers of puppies
- sales records
Housekeeping including:
- accommodation
- construction
- cleanliness and disinfection
- bedding
- exercise areas
- kitchen facilities
- first aid provision – human and animal
- food and water – storage and provision
- isolation facilities
- emergency procedures and fire prevention
Preparing the Bitch
No bitch should ever be breed before the 2, they are not physically mature yet. In the first 2 years of the dogs life they will be preparing for breeding so ensure you provide good quality nutrition, health care and training to ensure they are a good mother and produce a healthy litter. Another reason you must ensure your dog is over 2 years of old before breeding is health checks, you will need to ensure various health checks are carried out my a veterinary professional before you determine whether your dog is suitable for breeding. Please consult your vet for more information on the specific tests required.
Choosing a Stud Dog
Choosing the right sire for your litter is very important, and expert advice should be sought. The breeder of your bitch would be ideal for explaining what type of dog would be ideal and explaining this process to you.
Paperwork Required
Ensure you record the following information on file for each bitch and litter you produced:
Heat Record
- Name of bitch
- Litter Number (way to differentiate between litters at your kennel)
- Date of onset
- Interval
- Smear date and results
- Progesterone Test date and results
- Breeding dates and comments on breeding
- Palpitation dates and results
- Ultrasound date and results
- X-ray date and results
- Notes on pregnancy
- Track weight gain weekly
- Track temperature from day 58-65, 3 times daily
- Date and time whelping began
- Date and time whelping ended
- Notes on whelping
Litter Record
- Breed
- Registered name and KC number of dam
- Registered name and KC number of sire
- Sire's owner's name
- Date mated
- Date litter whelped
- Number of male puppies born
- Number of female puppies born
- KC Litter Number
- Sex, Colour/Markings, Puppy ID number, Date Sold, Date Died, Name and address of person to whom sold, Dates when following paperwork was supplied: registration application or certificate and bill of sale; name and KC number of puppy.
Additional Litter Information
- Time each puppy was born
- Ribbon colour or other identifying mark
- Colour of puppy
- Sex
- Weight at birth
- Length at birth
- A description of any problems
- Whelping date
- Sire and Dam
- Time whelping started and ended
- Notes on whelping
Puppy Record
- Ribbon colour
- Call Name
- Registered Name
- Sex
- Colour
- KC Litter No.
- KC Registration No.
- Date of Birth
- Sire and Dam
- Weight at Birth and when sold
- Vaccinations Given (Date and Type)
- Owner (include address and telephone numbers)
- Date sold
- Conditions of sale
- Price
- Notes on Development and Temperament
- On the back of this form, track the weight of the puppies daily until they are three weeks old and then weekly thereafter.
Litter Registration Application
Its very important to register you new litter with KC (The Kennel Club), this application should be completed as soon as the puppies are whelped so you can deliver the correct paperwork to the puppy buyers.
Puppy Registration Forms
Once your litter has been registered you will receive a registration form so the puppy buyers can register their puppies with the KC.


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Miami Heat guard Dwyane Wade and NBA commissioner David Stern had a contentious moment during Friday's NBA labor meeting.
Stern made a comment and pointed at Wade, who took offense to the move and felt like Stern was belittling him. Wade responded by yelling his rebuttal at Stern, ESPN.com learned from a source close to the situation.
Miami Heat guard Dwyane Wade had an exchange of terse words with NBA commissioner David Stern.
"You're not pointing your finger at me," Wade said, sources told ESPN. "I'm not your child."
Moments after the blow-up, Roger Goodell was spotted eaves-dropping and witness heard him mutter:
"He's losing his mind and I'm reaping all the benefits."


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Utah head coach Kyle Whittingham was reluctant to say who might have separated themselves from their teammates at Friday's scrimmage. (Story here.)
He wants to look at the tape. He wants to get it right.
But that's no fun, is it?
Utah head coach Kyle Whittingham was reluctant to say who might have separated themselves from their teammates at Friday's scrimmage. (Story here.)
He wants to look at the tape. He wants to get it right.
But that's no fun, is it?
With the Jeremiah Poutasi-sized caveat that this is the opinion of one unathletic sports reporter at The Tribune, here are a few guys who may have moved up the board with their play Friday and in recent practices.
Redshirt freshman quarterback Conner Manning
Last year, coaches weighed pulling Manning's redshirt and said, at least initially, that they felt junior Adam Schulz gave them the best chance to win. Is that still the case? If you judged from Friday's performances alone, no. Manning not only connected on a higher percentage of his passes 9-of-16 compared to 1-of-7 but among his incompletions were near-misses that might have been touchdowns.
Sophomore running back Marcus Sanders-Williams (see notes below) and junior Bubba Poole
It might seem odd to say that Poole's stock could rise when he's the No. 1 guy, but some of the shine has come off emerging threats Devontae Booker (junior) and Troy McCormick (redshirt freshman). They haven't fared poorly, either, but the consistent Poole outdid them Friday with 10 carries for 45 yards and three catches for 31 yards and a touchdown.
Offensive lineman ???
This is quite the puzzle. It's a safe bet that senior Junior Salt, a nonparticpant this spring, will start at guard in the fall. It's a safe bet that Jeremiah Poutasi will start SOMEWHERE in the fall (he was at left tackle Friday after playing left guard and right tackle earlier this spring). It seems a less-safe-but-still-fairly-good bet that sophomore Isaac Asiata will be at right tackle or guard and that junior Siaosi Aiono will start at center. So that leaves about 10 guys vying for the fifth position but very little insight into what that position may be. Your guess is as good as mine.
Senior wide receiver Andre Lewis (see notes below) and sophomore wide receiver Dominique Hatfield
He only had two catches for 23 yards, but the 5-foot-10, 170-pound Hatfield caught a Conner Manning pass and drove his legs forward with defenders in tow like a Radio Flyer. Hatfield has sticky hands and elite speed and has regularly shone among the brightest even with stiff competition from sophomores Brian Allen and Delshawn McClellon.
Sophomore left end Pita Taumoepenu
He's undersized at 6-foot-1, 225 pounds, but it often seems like Taumoepenu is the one with the unfair advantage. Defensive coordinator Kalani Sitake said after Friday's scrimmage that Taumoepenu's pass rush is "explosive," and it's just a matter of rounding out his game before making him an every-down player.
Sophomore stud linebacker Uaea Masina
Since Jacoby Hale tore his ACL with Jason Whittingham also missing time this spring the former Brighton standout has drawn praise from coaches for making the most of his opportunity to play frequently with the No. 1 defense. At 217, it's possible he could stand to tack on a few pounds, but he's got time to do it.
Junior free safety Charles Henderson
All signs point to Brian Blechen, Eric Rowe and Tevin Carter splitting up the safety spots in the fall. Still, Henderson has been the source of some this spring's hardest hits and he has a nose for the ball that hasn't gone ignored. Sitake said Henderson was one of his best players Friday.
OFFENSIVE HIGHLIGHT • No. 86 had everybody fumbling around for their rosters Friday. Sophomore tight end Ken Hampel may be buried deep in the tight end rotation, but he had two of the day's best plays. First the 250-pounder caught a Brandon Cox pass on a post route and rumbled forward, dragging defenders down the field with him, and then he ripped a 50/50 Jason Thompson toss out of the hands of freshman corner Jerry Padilla.
DEFENSIVE HIGHLIGHT • There weren't a ton. But the nod goes to converted as of Monday defensive end Greg Reese for the senior's recovery of a Cox fumble
STANDOUTS • Wilson and Taumoepenu are the obvious picks, but here are some runners-up: Sophomore cornerback Justin Thomas obliterated McCormick as the latter attempted to catch a swing pass. On offense, 222-pound sophomore running back Marcus Sanders-Williams plowed through defenders and then showcased his 4.43 speed on a 39-yard run.
KEEP AN EYE ON • Sophomore cornerback Hipolito Corporan played safety Friday, a move that Whittingham said was made "due to a lack of healthy bodies" at the position. Whittingham said it's not necessarily permanent, but he has the size (6'1, 178 pounds) for the position and will spend the final six practices of spring playing centerfield.
SCARY MOMENT • Senior receiver Andre Lewis has been highlighted in a few of these practice reports now, but Friday he was the recipient of a hard, low tackle from senior corner Wykie Freeman that made him writhe in pain as 300 or so people at Rice-Eccles Stadium went silent. Luckily for the Utes, Lewis was eventually able to stand up and walk off the field on his own power.
Matthew Piper
Twitter: @matthew_piper


text:
I've never tried Powerline kits, but I am interested in trying them out.. I'm really curious actually as to how well they work, I've though about buying some just to test it out, but just never have… Anyway, brite-view just came out with a new Powerline kit called the LinkE which is supposedly the world's smallest ethernet powerline adapters. It's a two part kit consisting of one box that gets plugged into your wall outlet near your modem which then gets plugged into it, the second part is a four port adapter that gets plugged in elsewhere in your house which then you can plug devices into and have much faster speed than you would over wireless. Don't know, has anyone out there tried Powerline? What do you think of it?
ZINNET® had brite-View(TM) CinemaTube customers in mind with today's launch of the "LinkE," a HomePlug AV-compliant powerline communication kit containing two Ethernet adapters. CinemaTube owners may assume that when their TV and CinemaTube aren't near a modem or router for a wired network connection, the CinemaTube can't stream Blu-ray quality movies over the WiFi network. This is not necessarily so.
With the "LinkE" powerline communication kit, consumers can take full advantage of their high-speed internet at up to 200Mbps. To use the "LinkE," first plug the world's smallest one-port Ethernet bridge (BV-210) into a wall socket and connect it to a modem with an Ethernet cable. Then plug the kit's four-port switch (BV-254) into a wall socket in another room, even on another floor where users want to network multiple devices such as the CinemaTube, Tivo, XBOX and even a Skype VoIP phone, using existing electrical wiring. The "LinkE" kit eliminates any WiFi dead zones in the house, and fits into the wall sockets perfectly without blocking other outlets.
Simple, fast and secure, powerline technology is not as widespread as it deserves to be because it is considered expensive and a Wi-Fi router is considered a more economical way of sharing the network with the whole house.
According to Robert Lo, CEO of ZINNET® though, "A WiFi network can be fragile. Without proper setup or protection, nicer neighbors can steal your Internet access while the evil neighbors hack into your computers."
brite-View(TM) is now offering the "LinkE" kit at a promotional price of $89.99, and the "CinemaTube plus LinkE" bundle for $174.99 on http://www.brite-view.com/. LinkE is a secure and cost-effective digital home solution that allows you to stream high-definition entertainment and enjoy online gaming without latency.
About ZINNET®, Inc.
ZINNET®, Inc. is a subsidiary of Zinwell® Corporation, a publicly listed company in Taiwan.
Headquartered in Los Angeles, CA with the Internet Division located in San Jose, CA, ZINNET® designs and manufactures consumer electronics that turn your living room into a digital home, including HD multi-media player with download features, and wireless HD transmission solutions. ZINNET® provides rich internet-based multi-media user experiences for the twenty-first century. ZINNET's products are backed up by nearly 30 years of industry experience and the ground breaking innovation ability that flourished from it.
Web Site: http://www.brite-view.com/


text: Smell the Coffee: Almost isn't good enough By Karin Fuller August 10, 2013 Story Share CHARLESTON, W.Va. -- The only surprising part about last week's "Jeopardy!" drama was that it didn't happen sooner.For those who might have missed it, "Jeopardy!" host Alex Trebek was taken to task over controversy that erupted after a 12-year-old contestant on a special children's edition of the popular game show misspelled his answer during Final Jeopardy.The question was to name the document, which took effect in 1863, that Abraham Lincoln called "a fit and necessary war measure."The answer was the Emancipation Proclamation, but Thomas Hurley III, an eighth-grader from Newtown, Conn., spelled his response "emanciptation" and his answer was ruled incorrect.Even if Hurley had spelled the word correctly, he was still too far behind the leader to have won, so while his mistake ended up costing him $2,000, it was not what prevented him from being the champion. Still, that didn't stop the fury from being unleashed, with many claiming the boy had been cheated, including Hurley's parents.Many of those complaining said the spelling error wasn't that bad, that he was fairly close, that you could tell he knew the correct answer, so that should've been enough.In a country where every child gets a trophy, where everyone's a winner all the time, regardless of effort or skill, I suppose it shouldn't be surprising that we've reached a point where we're wanting to also reward "close enough."Some said Hurley clearly knew the correct answer, but not how to spell it, so it should've been accepted. But for the show to accept anything less than correct would mean that from that point forward, a barely recognizable or incomplete answer would have to be considered, with judges having to guess at the contestant's intent or depth of knowledge. Instead, the show's producers drew a line in the sand."If 'Jeopardy!' were to give credit for an incorrect response (however minor), the show would effectively penalize the other players," the producers of "Jeopardy!" wrote. "We love presenting young people as contestants on our show and make every effort to be fair and consistent in their treatment."I admire the show for not backing down. We aren't doing children a favor by making everything gentle and even. A child would be better prepared for a life that is seldom fair if they're taught from an early age how to handle -- with grace -- the hand that they're dealt.Feeling disappointed or even embarrassed by a loss is completely normal, but it's how the child is taught to deal with what happened that matters. Rather than wallow and complain, Hurley should've been advised to stand up straight, give himself a pat on the back for having been clever enough to qualify for the show, and then maybe been shown how to make a self-deprecating joke or two that would show his strength of character, and move on.If we continue treating everyone the same, regardless of effort or skill, we're going to have a seriously hot mess on our hands. We're going to be dealing with generations raised to believe they're entitled to much that they simply are not. An almost-right answer is not the same as the right one. A ball downed on the one-yard line is not a touchdown. Almost getting the job doesn't get one a paycheck. That's life.Some will probably disagree, claiming there's no harm in giving trophies or pretty certificates or more of "Jeopardy!'s" money to all, but what they're failing to see is that by rewarding everyone, the prize is devalued. How is one child's self-esteem more important than the other child's hard work and achievement? Rewarding the child who was wrong takes away from the one who was right.We need to stop looking at it as raising children. We're raising adults. If we want them to become functioning members of society, they need to learn how to win and how to lose. They need to be able to take criticism, cope with arbitrary decisions and handle setbacks with class. And they need to see that people who work hard to achieve are rewarded more than those who do not.Reach Karin Fuller via email at firstname.lastname@example.com. Story Share Story Share Trending Now 1 Deer in 8 WV counties found dead from EHD 2 Plans for all-ages venue rise in old bakery building 3 Body found in Kanawha River, near South Charleston 4 Judge gives water crisis settlement preliminary approval 5 Capito quiet on newest ACA repeal proposal; Manchin opposes Hall past: Fascinating history in Wood County Previous Story Tie one on: Scarves are an elegant accessory Next Story


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4th Annual Clemson Easter Bunny Run 5k & Half Marathon
Adventure Geek Productions
Saturday, April 7, 2012 from 8:00 AM to 12:30 PM (EDT)
Event Details
The 4th Annual Clemson Easter Bunny Run 5k and Half Marathon
Saturday April 7, 2012
South Carolina Botanical Gardens
Clemson University
150 Discovery Ln
Clemson SC 29634
Brief Description:
Come join us in an exciting morning of fun and fitness. Chase the crazy carrot with your speed enhancing bunny ears as you run the half marathon or 5k. A Crazy Carrot will lead you on a scenic course through Clemson and the surrounding area. If you find an Easter egg along the way, be sure to pick it up for a surprise at the finish. An Easter egg hunt will also take place on the house lawn at the Botanical Gardens for all who wish to participate.
Expect new, fun, and exciting prizes! All half marathon finishers will receive a metal medal for their efforts and participation.
Those running the half marathon will receive the full Clemson running experience. Our course starts off in the South Carolina Botanical Gardens, circles around the Universities campus, adjacent neighborhoods and dikes giving you an excellent opportunity to see our beautiful Lake Hartwell.
The 5k is a rolling course includes the South Carolina Botanical Gardens and tours through part of campus.
*The Half Marathon course features mixed terrain and is NOT wheelchair accessible.
Know thy course! The courses plotted out on MapMyRun.com provide awesome resources like a 3D Fly Over, turn by turn directions, and course elevation information.
Half Marathon Course: http://www.mapmyrun.com/routes/view/24674072
5k Course: http://www.mapmyrun.com/routes/view/24672770
All turns on the course will be marked with signs and colored duck tap arrows on the ground.
The Half Marathon Course will be marked with BRIGHT GREEN duct tape arrows.
The 5k Course will be marked with BRIGHT PINK duct tape arrows.
All major intersection will have police support. Both races will have a police lead and follow up vehicle.
The 5k will have a water/Gatorade/first aid stop half way through and at the finish. The 5k is run, jog, walk and stroller friendly but please leave the pooch at home.
The Half Marathon will have a water/Gatorade/first aid stop approximately every 2 miles along the course in areas with lower vehicle traffic. The half marathon event is a race with a cut off time. Race participants need to be past the mile 7 marker by 9:45am (1:45:00 into the race = 14:52min/mile pace) to continue on with the course.
Map of Half Marathon - Always study and know your race course!
Map of 5k
Course Map
Early Registration Half Marathon - $45
Early Registration 5k - $18
Pre-Registration Half Marathon - $55
Pre-Registration 5k - $20
Race day registration fees:
Half Marathon- $70
5k - $25
We will be accepting Donations via Check or Cash. Please make checks payable to Clemson Triathlon Club. All proceeds will help send the active members of the Triathlon Club to Collegiate Nationals in Alabama. Go Tigers!
Race Day Registration:
Race day registration will be available from 6:00-7:45am.
Race Day Registration Fees:
Half Marathon - $70
5k - $25
Participants who register on race day may not receive a t-shirt due to limited supply.
We have the ability to accept credit cards at the race site now!
6:00am - 7:30am Registration/Check-in
8:00am - Half Marathon Start
8:10am - 5k Start
9:30am - Easter Egg Hunt
9:45am - 5k Awards
11:00am - Half Marathon Awards
Pre-Race Course, Safety and Venue Announcements will be made at 6:45, 7:15 and 7:45.
You do not need to attend all three sessions but you need to attend at least one.
Sponsors and Volunteers:
For information on becoming a sponsor or volunteer please click here.
Current Sponsors:
Tiger Town Graphics
SC Botanical Gardens
University Baptist Church
Adventure Geek Productions
2011 - Overall Half Marathon Results
2011 - Age Group Half Marathon Results
2011 - Overall 5k Results
2011 - Age Group 5k Results
2011 Race Photos can be found at www.backprint.com/themurraygroup
2009 Pictures on Picasa.
Volunteer & Vendor Information:
Please contact Jefferson or Kristen (information below) to sign-up. Please arrive by 6:00am and plan to stay until 1:00pm.
Contact Information & Volunteers Opportunities:
Race Directors: Jefferson Nicholson, 864-247-6735, email@example.com
Kristen Thomas, 803-487-1052, firstname.lastname@example.com
Top 3 Male & Female
Top 3 Masters Male & Female
Age Group Top 3 Male & Female:
14 & under
Additional Information:
Race Proceeds will benefit the Clemson Triathlon Club
Refund Policy:
50% of your entry fee will be refunded until March 7, 2012 or until the race has reached capacity, whichever occurs first. No refunds will be issued for any reason after March 7, 2012 or after the race is full. Event entries are not transferable between persons, events or years. NO EXCEPTIONS will be made to this policy. By registering for this event you agree to these refund terms.
When & Where
South Carolina Botanical Gardens
150 Discovery Ln
Saturday, April 7, 2012 from 8:00 AM to 12:30 PM (EDT)
Add to my calendar


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A Serious and Funny Man
When I started Steve Martin's autobiography, Born Standing Up, I didn't expect to think it might be a good book for discussion. But when I finished it, getting misty on the Metro bus, I began to think otherwise.
Funnyman, comic, writer, playwright, and actor Steve Martin is as much a topic for discussion as any other subject. Born Standing Up finds Martin reliving his hungry days, 18 years of honing and perfecting his unique take on new comedy. From his upbringing in California in the 1950s, his fraught relationship with his father, and his life on the road, Martin shows us just how serious comedy can be and how much his rise cost him.
Years ago, I had seen his play, Picasso at the Lapin Agile, performed in a small Vermont theater house. At the time, I had no clue that the "wild and crazy guy" I knew from Saturday Night Live had writing talent up his sleeve.
Then novels like Shopgirl and The Pleasure of My Company started coming out.
But what really caused me first to take a second look at Steve Martin was an essay that he wrote about his father's death for The New Yorker. That article remains one of the most moving, emotional pieces about a father-son relationship that I had ever read, made more striking for the effort and release it obviously must have provided for the writer. Parts of this essay ended up incorporated into his memoir, but the essay is still more powerful to me.
It wasn't until I saw Jerry Seinfeld's 2002 documentary, Comedian, that I realized how much work and thinking is involved in comedy. I don't know why it took me so long to realize that is like any other art.
But Steve Martin grew up in a different time, and his brand of comedy took years to create and perfect. Martin also talks about how and why he left stand-up and got into filmmaking, and after following him on his arduous journey, you understand why.
Born Standing Up would be a wonderful book for discussion. It could be paired with one of Martin's novels or plays, or could be enhanced by listening to one of his comedy albums. You could discuss and watch Shopgirl, in which Martin also acts. The possibilities are limitless. And if your group chooses to do so, let me know how it goes.


text:
The Miami Heat front office is thanking its lucky stars that LeBron James and the rest of the team have great reflexes.
While reveling in his second of back-to-back championships on top of an open double-decker bus, James very nearly went face-first into a concrete overpass.
The glass-enclosed overpass had a ton of fans elated to catch a glimpse of their superstar, but none that warned him about the low clearance.
MORE: Parade pictures | Heat celebrate title with city of Miami
A bunch of really tall guys standing on top of a really tall bus on a parade route that has bridges with 14-foot clearances.
Did no one have the foresight to think this plan through entirely?
(First found on FTW)


text:
View all
Crossbows Recalled-Injury Hazard
Excalibur Crossbows Inc. has recently recalled about 1,000 crossbows due to an injury hazard.
About the Crossbow Recall
According to the Consumer Product Safety Commission (CPSC), Excalibur has received one report of the crossbow firing unexpectedly.
The crossbow could pose as an injury hazard to consumers. CPSC recommends that consumers stop using the recalled crossbows and immediately call Excalibur for a free inspection and repair.
Consumers can also call Excalibur for shipping instructions to Excalibur for a free repair and free shipping.
Description of Recalled Crossbows
- Crossbows being recalled are the Excalibur Matrix Mega 405 crossbows. On the left side of the stock there is a printed label that says "Matrix 405".
- The crossbow measures 36 inches long and was sold in a mossy oak tree stand camouflage pattern. It is sold individually or in addition to four-arrow quiver and bracket, four arrows, and four 150-grain field points and rope-cocking aid.
- Crossbows were sold at sporting goods nationwide from February 2014 through March 2014 for $1000 for the crossbow and $1400 when sold as a package with crossbow and additional equipment.


text:
DrivingTest.ca is an online driving test practice site that serves all provinces and territories in Canada. Advance through the practice tests at your own pace on any computer, tablet or mobile phone with internet access. While these tests are not the same as the actual ones you will be taking, they can give you a good idea of what to expect. It is all FREE!
Question 1
School vehicles turning left ahead
School crosswalk sign
Caution: school bus crossing
Regulatory sign
Question 2
Construction zone ahead: slow down
Bridge or Viaduct ahead
Bump or uneven pavement on the road ahead: slow down
Construction zone
Question 3
Road turns right, then left
Pass other traffic on the right
Winding road ahead
Keep to the right of the traffic island
Question 4
Do not turn left at the intersection
Hidden intersection ahead
Right turn only at the intersection
Traffic island ahead
Question 5
Pedestrian control sign
School zone: watch for children playing
Destination board
Provincial park notification
Question 6
Railway crossing ahead
Bump 120 meters ahead
Traffic lights ahead: slow down
Stop 150 meters ahead
Question 7
Pavement is slippery when wet: slow down and drive with caution
Bumpy road ahead
Winding road ahead
Danger: hunting area ahead
Question 8
Hospital zone ahead: yield to oncoming traffic
Pedestrian crosswalk ahead: yield to pedestrians
4-way intersection ahead: through traffic may proceed
Railway crossing ahead: be alert for trains
Question 9
Road work ahead
Flagman on duty ahead
Pedestrian control sign
School area ahead
Question 10
Direction sign for children
School zone
Children playing in residential area
Playground area sign
Question 11
Pedestrian crossing: be prepared to stop and yield right-of-way to pedestrians.
Pedestrians or vehicles may not enter the roadway
Railroad crossing ahead
Deer crossing ahead
Question 12
Route for large trucks
Permissive sign
Traffic circle ahead
No trucks
Question 13
Stop sign ahead
Slow-moving vehicle (travelling less than 40 km/h)
Dead-end street ahead
Yield right-of-way to vehicle ahead
Question 14
Stop only if other vehicles are approaching
Drivers must come to a complete stop
Bump 150 meters ahead
Stop 150 meters ahead
Question 15
Road slippery when wet
Narrow road ahead
Winding road ahead
Hidden intersection ahead
Question 16
Keep left
Do not enter this road
Do not pass
Keep right
Question 17
Speed limit changes ahead
End of 50 km per hour zone
Rural school zone ahead
Maximum speed 50 km/h from this sign to the next
Question 18
Crosswalk ahead
Sharp turn guides
Exit to the right
Winding road ahead
Question 19
Merge Ahead
Two lanes ahead: both lanes travel in the same direction
Road turns to the left
Road turns to the right
Question 20
Steep hill ahead: be prepared to use a lower gear if necessary
Road closed ahead due to fallen rocks
Watch for fallen rock: be prepared to avoid rocks that may have fallen on the road
Construction zone ahead
Question 21
Divided highway ends
Road under construction
Narrow bridge ahead
Divided roadway ahead
Question 22
Yield: give the right-of-way
Railway crossing ahead
Oncoming traffic must stop
School zone ahead
Question 23
Right turn lane
No parking from arrows to corner
Left turn lane
Danger ahead: veer to the right
Question 24
Winding road ahead
Keep right
Sharp turn or bend in the road in the direction of the arrow
Danger: road ends
Question 25
Steep hill ahead
Hidden intersection ahead
Right lane ends ahead
Pedestrian crossing ahead
Question 26
U-turn prohibited
Road slippery when wet
Left turn prohibited
Right turn prohibited
Question 27
Local traffic only
Traffic circle ahead
Prohibited sign
Entry to roadway prohibited
Question 28
One-way street ahead
Hidden intersection ahead
Pavement/Road narrows ahead
Divided highway ahead
Question 29
Pavement ends 12 feet ahead
Safety check ahead
Bumpy road ahead
Steep hill ahead: be prepared to use a lower gear if necessary
Question 30
No stopping in the area between the signs
Approaching vehicles must stop, then may proceed with caution
No entry into intersection
There are no stop signs in the direction of the arrows: proceed with caution
It is recommended that you take a practice test before you take the actual written test. Please note, however, that this is not the actual test, and you will not receive your license if you pass this sample practice test. If you're getting your driver's licence, you've come to the right place.
DRIVINGTEST.CA based solely on the idea of providing a simpler and more user friendly approach to the world of driving. Our goal is not only to teach people how to safely operate a motor vehicle, but also to give the best possible online training for becoming smart & skillful drivers in this highly motorized age.
This online practice test, with multiple choice questions, is intended to help you prepare for the PEI Class 7 (Stage 1) learner driver's test that is required before you can get your driver's license. This free practice written driving test is a lesson that is essential for getting your PEI driver's license at your local licensing office. Please note, however, that this is not the actual test, and you will not receive your license if you pass this practice test. It is all FREE!
Sometimes all it takes to succeed at something is good coaching. This applies to athletes as much as it applies to future drivers. The time spent preparing for the Driver's License written exam is the most crucial to take advantage of properly. With a little help, a perfect mark on the exam is possible, even if it takes more than one try.


text: I have spent most of my life in martial arts. The past 15 years I have placed my focus on "Self Defense". I have studied, trained in, and taught KRAV MAGA, the Israeli Self Defense System used by the IDF and Israeli Law Enforcement. Having said all of that, it's my expert opinion that one of the best tools to defend the lives of you and your loved ones is a GUN!