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As another sports season has ended, it's time to announce the spring Athletes of the Year.
As another sports season has ended, it's time to announce the spring Athletes of the Year. Here are the honorees:Alec Tuohy BASEBALL
Tuohy pitched Gahanna to a win in a Division I district final in 2011 but dealt with a learning curve a year ago while adjusting to being the Lions' top starting pitcher and shortstop.
The senior thrived in both roles this season, having a team-high .454 batting average and going 9-2 on the mound with a 2.37 ERA and 102 strikeouts in 73 2/3 innings. He was named district Player of the Year, second-team all-state and first-team all-OCC-Ohio Division.
Tuohy struck out 14 in a 7-2 win over Upper Arlington in a district semifinal May 20, breaking the program record for most strikeouts in a seven-inning game, and he homered and had six strikeouts in six innings in a 10-1 win over Marysville in a regional semifinal May 30.
The University at Buffalo recruit won 19 games over the last three seasons. This season, the Lions went 22-10 and reached a state semifinal for the first time.
"He just took his game to the next level and just had a great year," Gahanna coach Mike Shade said. "This year in the tournament, he went out and basically just took over."Emma Cornell SOFTBALL
This season, Cornell helped lead Grove City to the second of back-to-back Division I district and OCC-Ohio Division championships.
The junior left-handed pitcher and designated player went 17-4 in the circle with a 1.14 ERA and 130 strikeouts and only 22 walks in 112 2/3 innings. At the plate, she had a .467 batting average with 25 RBI, 28 runs and seven doubles.
"Emma did an outstanding job," Greyhounds coach Ted Williams said. "She kept us in every ballgame we were in. ... She hit the ball well on top of pitching. You can't ask for anymore from a young lady."
Cornell was named first-team all-state and all-district and league Player of the Year, as the Greyhounds finished 27-5 and reached a regional final before losing to eventual state champion North Canton Hoover 4-1 on June 1 at Ohio State.
"I gained a couple mph on my fastball," said Cornell, who said her top speed this season was approximately 63 mph. "I feel like I have progressed a lot this year, mentally and physically, playing ball."Jack Warmolts BOYS LACROSSE
Warmolts, a 6-foot-3, 220-pound senior goalkeeper, averaged eight saves per game as Upper Arlington won its second consecutive Division I state championship, third in four years and 14th overall.
A three-year starter, Warmolts earned first-team all-state, all-Central Region and all-OCC-Buckeye Division honors for the second consecutive year. He also was a first-team All-American this season along with two teammates, midfielder Gus Ackley and defender Jake Nolan, as the Golden Bears finished 20-2 overall and 6-0 in the OCC-Buckeye.
Warmolts, whose only shutout this season came in a 6-0 win over Thomas Worthington on April 13, will play at the U.S. Air Force Academy.
"Jack's a class act, from the standpoint of being a great kid before he's a great ballplayer," UA coach Ted Wolford said. "He's worked so hard all the way through toward having the ability to be a leader, and now all the way through to where he's an All-American. That's something you feel really good about. ... He's a solid, solid kid."Elaina Enich GIRLS LACROSSE
Enich, a senior attacker, scored 87 goals and had six assists this season to help lead New Albany to the Division II state championship.
The University of Cincinnati signee scored a program-record nine goals in a 19-12 win over Olentangy Liberty on April 18 and tied the record in an 18-6 win over Granville on May 16.
Enich had six goals in a 19-17 win over Shaker Heights Hathaway Brown in a state semifinal May 31 and scored four in an 11-10 win over Worthington Kilbourne in the state final June 1.
Enich, who missed the end of her junior season with a torn anterior cruciate ligament, was named first-team All-American as well as first-team all-state, all-South/Central Region and all-OCC-Buckeye Division. She is the first two-time All-American in program history.
"There was never a doubt that she would return to play her senior year and continue to be a dominant player," Eagles coach Dave Ferguson said. "She was regarded as one of the most dangerous offensive players in the state by many coaches."Stu Little BOYS TENNIS
An Upper Arlington senior and University of Pennsylvania recruit, Little placed third in singles in the Division I state tournament and led the Golden Bears to their third Ohio Tennis Coaches Association team championship in four years.
A four-time state qualifier who teamed with 2011 graduate Billy Weldon to win state doubles titles in 2010 and 2011, Little lost to Cuyahoga Falls Walsh Jesuit's Aaron Sandberg 4-6, 6-1, 7-6 (7-2) in a state semifinal June 1 at Ohio State before defeating Gahanna's Canyon Teague 6-3, 6-3 in the third-place match later that day.
Despite his achievements, Little, who lost in a state quarterfinal in singles in 2012, said he will rue the state titles he didn't win.
"That's just the kind of kid Stu is," UA coach Amos Allison said. "Years down the road, he'll look back and still kick himself for a few points he didn't get in his semifinal match. At the same time, he'll realize what he did and what he was a part of. It's one of the most impressive things a kid can do."Desmond Palmer BOYS TRACK & FIELD
Pickerington North might have been the deepest boys team in central Ohio this season, but the Panthers wouldn't have earned their second consecutive Division I state runner-up finish without Palmer.
The senior and University of Pittsburgh signee helped score 40 of North's 47 points in the state meet June 7 and 8 at Ohio State, as the Panthers placed second to Lakewood St. Edward (62) for the second year in a row.
Palmer won state championships in the 110-meter hurdles (14.03 seconds) and 300 hurdles (36.31) and anchored the winning 800 relay (1:26.14) and the first-place 1,600 relay (3:14.73).
On June 20, Palmer, who also won district and regional titles in all four events, was named Gatorade Ohio Boys Track and Field Athlete of the Year. His time in the 300 hurdles at state ranked third in the nation in 2013 at the time the Gatorade honor was announced.
"(State) was one of those meets where Desmond said, 'Jump on my back,' and we got four first-place finishes," North coach Dave Spring said.Aaliyah Barnes GIRLS TRACK & FIELD
Barnes believes she's ready to compete at the collegiate level given her performance in the Division II state meet June 7 and 8 at Ohio State.
The Eastmoor Academy senior qualified for state in three individual events and on a relay and elected to compete in all four events. She finished the meet with two first-place finishes, a second-place finish and a third-place finish, scoring 34 points as the Warriors won the team title.
Barnes has signed with Ohio State.
"Although I didn't get first in everything, I'm so proud of myself that I even (competed in three individual events at state)," she said. "This will get me ready for next year."
At regional May 30 and June 1 at Welcome Stadium in Dayton, Barnes won the 100 meters (12.51 seconds) and 200 (25.36), placed second in the 400 (56.44) and was on the winning 800 relay (1:41.2).
At state, she won the 100 (11.9), placed second in the 400 (55.77) and third in the 200 (24.7) and was on the winning 800 relay (1:40.38).
Barnes finished her prep career with seven state titles.Jarrod Kelso BOYS VOLLEYBALL
Kelso, a 6-foot-6 middle hitter, helped lead Hilliard Darby to its first Division I state championship.
In 26 matches, the senior had 259 kills, 59 blocks and a .657 hitting percentage to lead the Panthers, who finished 24-2 and defeated Cincinnati Elder 25-17, 25-15, 25-27, 25-23 in the state final May 26 at Cuyahoga Falls Walsh Jesuit.
Kelso was named state and East Region Player of the Year in Division I and the OCC-Ohio Division Player of the Year. As a junior, he was second-team all-state and first-team all-region.
"Jarrod Kelso was our best offensive player (in the 2012) season and he had a .415 hitting percentage," Darby coach Bryan Webb said. "He's worked hard on his craft and he (was) at .600 this year, which is almost unheard of. He (was) literally unstoppable, especially in the state tournament."
Kelso has committed to play for Quincy University in Illinois. Quincy is one of eight teams in the Midwestern Intercollegiate Volleyball Association, which also includes Ohio State.
-From staff reports


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Method, instruments, and kit for autologous transplantation
Method for the effective transplantation of chondrocytes/cartilage to an articular joint surface defect is taught, as well as a description of certain instruments and kit for practicing the invention.
This application is a continuation of U.S. patent application Ser. No. 08/857,090, filed May 15, 1997 (now U.S. Pat. No. 5,989,269), which is a continuation-in-part of U.S. patent application Ser. No. 08/704,891, filed Aug. 30, 1996 (now U.S. Pat. No. 5,759,190), both of which are hereby incorporated by reference in their entirety.FIELD OF THE INVENTION
The instant invention concerns the field of chondrocyte transplantation, bone and cartilage grafting, healing, joint repair and the prevention of arthritic pathologies. In particular methods for the preparation of the graft site, instruments for such preparation and for the autologous transplantation of cells to the prepared graft site.BACKGROUND OF THE INVENTION
More than 500,000 arthroplastic procedures and total joint replacements are performed each year in the United States. Approximately the same number of similar procedures are performed in Europe. Included in these numbers are about 90,000 total-knee replacements and around 50,000 procedures to repair defects in the knee per year in Europe. The number of procedures are essentially the same in the U.S. (In: Praemer A., Furner S., Rice, D. P., Musculoskeletal conditions in the United States, American Academy of Orthopaedic Surgeons, Park Ridge, Ill., 1992, 125). A method for regeneration-treatment of cartilage would be most useful, and could be performed at an earlier stage of joint damage, thus reducing the number of patients needing artificial joint replacement surgery. With such preventative methods of treatment, the number of patients developing osteoarthritis would also decrease.
Techniques used for resurfacing the cartilage structure in joints have mainly attempted to induce the repair of cartilage using subchondral drilling, abrasion and other methods whereby there is excision of diseased cartilage and subchondral bone, leaving vascularized cancellous bone exposed (Insall, J., Clin. Orthop. 1974,101,61; Ficat R. P. et al, Clin Orthop. 1979, 144, 74; Johnson L. L., In: Operative Arthroscopy, McGinty J. B., Ed., Raven Press, New York, 1991, 341).
Coon and Cahn (Science 1966, 153, 1116) described a technique for the cultivation of cartilage synthesizing cells from chick embryo somites. Later Cahn and Lasher (PNAS USA 1967, 58, 1131) used the system for analysis of the involvement of DNA synthesis as a prerequisite for cartilage differentiation. Chondrocytes respond to both EFG and FGF by growth (Gospodarowicz and Mescher, J. Cell Physiology 1977, 93, 117), but ultimately lose their differentiated function (Benya et al., Cell 1978, 15, 1313). Methods for growing chondrocytes were described and are principally being used with minor adjustments by Brittberg, M. et al. (New Engl. J. Med. 1994, 331, 889). Cells grown using these methods were used as autologous transplants into knee joints of patients. Additionally, Kolettas et al. (J. Cell Science 1995, 108, 1991) examined the expression of cartilage-specific molecules such as collagens and proteoglycans under prolonged cell culturing. They found that despite morphological changes during culturing in monolayer cultures (Aulthouse, A. et al., In Vitro Cell Dev. Biol., 1989,25,659; Archer, C. et al., J. Cell Sci. 1990,97,361; Hänselmann, H. et al., J. Cell Sci. 1994,107,17; Bonaventure, J. et al., Exp. Cell Res. 1994,212,97), when compared to suspension cultures grown over agarose gels, alginate beads or as spinner cultures (retaining a round cell morphology) tested by various scientists did not change the chondrocyte—expressed markers such as types II and IX collagens and the large aggregating proteoglycans, aggrecan, versican and link protein did not change (Kolettas, E. et al., J. Cell Science 1995,108,1991).
The articular chondrocytes are specialized mesenchymal derived cells found exclusively in cartilage. Cartilage is an avascular tissue whose physical properties depend on the extracellular matrix produced by the chondrocytes. During endochondral ossification chondrocytes undergo a maturation leading to cellular hypertrophy, characterized by the onset of expression of type X collagen (Upholt, W. B. and Olsen, R. R., In: Cartilage Molecular Aspects (Hall, B & Newman, S, Eds.) CRC Boca Raton 1991, 43; Reichenberger, E. et al., Dev. Biol. 1991, 148, 562; Kirsch, T. et al., Differentiation, 1992, 52, 89; Stephens, M. et al., J. Cell Sci. 1993, 103, 1111).
Excessive degredation of type II collagen in the outer layers or articular surfaces of joints is also caused by osteoarthritis. The collagen network is accordingly weakened and subsequently develops fibrillation whereby matrix substances such as proteoglycans are lost and eventually displaced entirely. Such fibrillation of weakened osteoarthritic cartilage can reach down to the calcified cartilage and into the subchondral bone (Kempson, G. E. et al., Biochim. Biophys. Acta 1976, 428, 741; Roth, V. and Mow, V. C., J. Bone Joint Surgery, 1980, 62A, 1102; Woo, S. L.-Y. et al., in Handbook of Bioengineering (R. Skalak and S. Chien eds.), McGraw-Hill, New York, 1987, pp. 4.1-4.44).
Descriptions of the basic development, histological and microscopic anatomy of bone, cartilage and other such connective tissues can be found for example in Wheater, Burkitt and Daniels, Functional Histology, 2nd Edition, (Churchill Livingstone, London, 1987, Chp. 4). Descriptions of the basic histological anatomy of defects in bone, cartilage and other connective tissue can be found for example in Wheater, Burkitt, Stevens and Lowe, Basic Histopathology (Churchill Livingstone, London, 1985, Chp. 21).
Despite the advances in cultivating chondrocytes, and manipulating bone and cartilage, there has not been great success with the attempts to transplant cartilage or chondrocytes for the repair of damaged articulating surfaces. The teachings of the instant invention provide for effective, and efficient means of promoting the transplantation of cartilage and/or chondrocytes into a defect in an articulating joint or other cartilage covered bone surface, whereby cartilage is regenerated to fix the defect. The instant invention also provides for surgical instruments which are designed prepare the graft site so as to facilitate the efficient integration of grafted material to the graft site.BRIEF SUMMARY OF THE INVENTION
The instant invention provides a method for the effective treatment of articulating joint surface cartilage by the transplantation of chondrocytes in a suitable matrix, to a surface to be treated, with a hemostatic barrier and a cell-free covering-patch comprising; first placing a hemostatic barrier proximal to the surface to be treated, placing chondrocytes in a suitable matrix upon the surface to be treated distal to the hemostatic barrier, covering the surface to be treated with a cell-free covering-patch. A hemostatic barrier, as will be further described below, is a barrier which inhibits the penetration of vascularizing cells and tissue into the grafted material. In particular, the instant method provides for a hemostatic barrier that is a resorbable, semi-permeable material which inhibits or prohibits vascular infiltration through the barrier. In one embodiment the hemostatic barrier contains collagen. Cell-free, is used herein as in the art, and means a material that is substantially free from intact cells which are capable of further cell division, promulgation or biological activty. In a preferred embodiment, a cell-free material is free from all intact nucleated cells. In one embodiment, the instant method encompasses the use of a cell-free covering patch which contains a semi-permeable collagen matrix. In one preferred embodiment of the method, the porous surface of the cell-free covering-patch is directed towards the implant material.
The instant invention further provides for the autologous transplantation of collagen or chondrocytes to a graft site, wherein the graft site has first been prepared by surgical manipulation to better accept the grafted material. In one embodiment, the graft site is sculpted such that the walls of the graft site are contoured in an undulating pattern such that the grafted material, when placed within the graft site and expanded to contact the graft site wall, there will be resistance against removal or expulsion of the entire graft from the graft site. The instant invention further provides for surgical instruments designed to sculpt the graft site as taught by the method of the invention.
The invention further provides for a kit for cartilage and/or chondrocyte transplantation onto the surface of an articular joint wherein said kit comprises a hemostatic barrier, cell-free semi-permeable covering-patch, and organic glue. In a further embodiment, the kit can optionally further provide one or more surgical instruments which can be used to sculpt the graft site in accordance with the methods of the instant invention.BRIEF DESCRIPTION OF THE DRAWINGS
The present invention will be better understood by examining the following figures which illustrate certain properties of the instant invention wherein:
FIG. 1A is a drawing showing a typical articulating end of a bone. Typically, the bone material is covered on the articulating surface with a cartilaginous cap (shown by cross-hatching).
FIG. 1B shows an example of where a defect or injury to the cartilaginous cap occurs (gap in the cross-hatching), and such a defect can be treated directly, enlarged slightly, or sculpted to accept the grafted material by surgical procedures prior to treatment.
FIG. 1C shows how the hemostatic barrier (solid black, numbered 1) is placed within the defect in the cartilage cap to inhibit or prevent vascularization into regenerating cartilage, from the underlying bone. The chondrocytes to be implanted into the defect cavity are then layered on top of the hemostatic barrier.
FIG. 2 is a drawing showing the treated defect (gap in cross-hatched area) in the cartilaginous cap (cross-hatched area) covered by a cell-free semi-permeable material (solid black, numbered 2) which is used to form a cap/patch or bandage over the defect site. This cap is fixed in place, either sutured to the edge of the cavity into healthy cartilage, or otherwise attached. This cap is covering the defective area of the joint into which the cultured chondrocytes/cartilage transplant has been placed.
FIG. 3A is a diagram illustrating the differential response to compression and shearing forces by harder and softer cartilage with subsequent zone of demarcation.
FIG. 3B illustrates the graft site, after the defect has been sculpted to have undulating walls.
FIG. 3C illustrates the sculpted graft site with the hemostatic barrier (1), transplanted material (3), and cell-free covering-patch (2) in place within the articular surface cartilage (4).
FIG. 4A illustrates one embodiment of the surgical device of the instant invention showing cutting teeth (5) and protruding placement pin (6).
FIG. 4B illustrates a second embodiment of the surgical device of the instant invention.
FIG. 5 is a diagram illustrating the modified differential response to compression and shearing forces by harder cartilage and softer cartilage after sculpting the graft site.
FIG. 6A is an MRI image of a pig knee showing cartilage defect in left (medial) condyle.
FIG. 6B is an MRI image of the same pig knee three months after treatment.DETAILED DESCRIPTION OF THE INVENTION
This invention concerns the use of certain products that inhibit the formation of vascular tissue, for instance such as capillary loops projecting into the cartilage being established, during the process of autologous transplantation of chondrocytes into defects in the cartilage. The formation of vascular tissue from the underlying bone will tend to project into the new cartilage to be formed leading to appearance of cells other than the mesenchymal specialized chondrocytes desired.
The contaminating cells introduced by the vascularization may give rise to encroachment and over-growth into the cartilage to be formed by the implanted chondrocytes. One of the types of commercial products which can be used in this invention is Surgicel® (Ethicon Ltd., UK) which is absorbable after a period of 7-14 days. The use of this material in the method of the instant invention is contrary to the normal use of a hemostatic device, such as Surgicel® as it is described in the package insert from Ethicon Ltd.
Surprisingly, we have found that in a situation where you wish to inhibit re-vascularization into cartilage, a hemostatic material material will act like a gel-like artificial coagulate. If red blood cells should be present within the full-thickness defect of articular cartilage that is capped by such a hemostatic barrier, these blood cells will be chemically changed to hematin, and thus rendered unable to induce vascular growth. Thus a hemostatic product used as a re-vascularization inhibitory barrier with or without fibrin adhesives, such as for example the Surgicel®, is effective for the envisioned method as taught by the instant invention. Another part of this invention is the use of a cell-free component, that is used as a patch covering the defective area of the joint into which the cultured chondrocytes/cartilage are being transplanted, using autologous chondrocytes for the transplantation. The method of the invention also contemplates the use of suitable allogenic chondrocytes or xenogenic chondrocytes for the repair of a cartilage defect.
Thus the instant invention teaches methods for effective repair or treatment of cartilage defects in articular joint bone surfaces which comprises administering an agent or device to block vascular invasion into the cartilage site to be repaired, and also providing for a cell-free barrier which will isolate the repair site and keep transplanted cells in place. Thus the instant invention also provides for a kit comprising a hemostatic barrier component for insertion into the site to be repaired, such that there is effective inhibition of vascularization into the site to be repaired; and once the chondrocytes to be transplanted are placed into the site to be repaired, a cell-free semi-permeable barrier is capped over the repair site such that the transplanted chondrocytes are held in place, but are still able to gain access to nutrients.
Certain aspects of the invention have been exemplified using an in vitro system to study the behavior of the chondrocytes when in contact with a certain product or a combination of certain products that inhibit the formation of vascular tissue. This in vitro testing predicts the ability of certain tested materials to inhibit vascularization, as will occur in vivo where capillary loops project into the cartilage being established during the process of autologous transplantation of chondrocytes into defects in the cartilage.
Suitable hemostatic products will be characterized by having the ability to inhibit the growth, or invasion of vascular tissue, osteocytes, fibroblasts etc. into the developing cartilage. A suitable hemostatic material will achieve the goal of the method of the instant invention in that vascular and cellular invasion into developing cartilage should be prevented in order to optimize the formation of cartilage and achieve repair of the full-thickness of any defects in the articular cartilage. Ideally, the hemostatic barrier will be stable for an extended period of time sufficient to allow for full cartilage repair, and then be able to be resorbed or otherwise broken down over time. One material identified as suitable is called Surgicel® W1912 (an absorbable hemostat containing oxidized regenerated sterile cellulose; Lot GG3DH, Ethicon Ltd. UK). Another example of a suitable material is BioGide® (a commercially available type I collagen matrix pad; Geistlich Söhne, Switzerland).
Suitable organic glue material can be found commercially, such as for example Tisseel® or Tissucol® (fibrin based adhesive; Immuno AG, Austria), Adhesive Protein (Cat. #A-2707, Sigma Chemical, USA), and Dow Corning Medical Adhesive B (Cat. #895-3, Dow Corning, USA).
The surgical instruments contemplated by the instant invention can be manufactured from metal and/or plastic suitable for making single-use disposable, or multi-use reusable surgical instruments. The cutting instrument may contain cutting teeth that are fully circular or flat, or anything in between. As cartilage is a relatively soft material it may be advantageous to manufacture hardened plastic cutting edges which will be able to sculpt cartilage without being able to damage bone. Such cutting instruments can be manufactured to incorporate openings for administration of fluid, suction removal of cutting debris and fluid, and fiber optic threads for illumination and visualization of the defect site.
Certain aspects of the instant invention may be better understood as illustrated by the following examples, which are meant by way of illustration and not limitation.EXAMPLE 1
In order for the Surgicel® to be used according to the invention for preventing development of blood vessels into autologous implanted cartilage or chondrocytes, Surgicel® was first treated with a fixative, such as glutaric aldehyde. Briefly, Surgicel® was treated with 0.6% glutaric aldehyde for 1 minute, followed by several washings to eliminate glutaric aldehyde residues that may otherwise be toxic to tissue. Alternatively, the Surgicel® was treated with the fibrin adhesive called Tisseel® prior to treatment with glutaric aldehyde as described in Example 2. It was found that the Surgicel® fixated for instance with a fixative such as glutaric aldehyde, washed with sterile physiological saline (0.9%) and stored in refrigerator, does not dissolve for 1 to 2 months. Generally, Surgical is resorbed in a period between 7 and 14 days. This time would be too short, because a longer time is needed in preventing the development of blood vessels or vascularization as such from the bone structure into the implanted cartilage before the implanted chondrocytes have grown into a solid cartilage layer getting its nutrition requirements from the neighboring cartilage. In other words sufficient inhibition of the vascularization is needed for a longer time such as for instance one month. Therefore, the product should not be absorbed significantly prior to that time. On the other hand resorption is needed eventually. Hence, the organic material used as an inhibiting barrier shall have these capabilities, and it ahs been found that the Surgicel® treated in this manner provides that function.EXAMPLE 2
The Surgicel® was also coated with an organic glue, in this example the glue used was Tisseel® but others can also be used. This product, together with the Surgicel® produces a useable barrier for the particular purpose of the invention. Any other hemostat or vascular inhibiting barrier could be used. The Tisseel® was mixed as described below. The Surgicel® was then coated with Tisseel® by spraying the Surgicel® material on both sides until soaked. The Tisseel® (fibrin glue) was then allowed to solidify at room temperature. Immediately prior to completed solidification, the coated Surgicel® was then placed in 0.6% glutaric aldehyde for 1 minute and then washed with sterile physiological (0.9%) saline. The pH was then adjusted by PBS and/or with NaOH until pH was stable at 7.2 to 7.4. Afterwards the thus treated Surgicel® was then washed in tissue culture medium such as minimum essential medium/F12 with 15 mM Hepes buffer.
As mentioned in this example we have used Tisseel® as the fibrin adhesive to coat the Surgicel®. Furthermore the fibrin adhesive or glue may also be applied directly on the bottom of the lesion towards the bone, on which the Surgicel® is glued. The in vitro system used, in lieu of in vivo testing, consisted of a NUNCLON™ Delta 6-well sterile disposable plate for cell research work (NUNC, InterMed, Roskilde, Denmark). Each well measures approximately 4 cm in diameter.
In the invention the fibrin adhesive can be any adhesive which together with the fibrin component will produce a glue that can be tolerated in humans (Ihara, N, et al., Burns Incl. Therm. Inj., 1984, 10, 396). The invention also anticipates any other glue component that can be used in lieu of the fibrin adhesive. In this invention we used Tisseel® or Tissucol® (Immuno AG, Vienna, Austria). The Tisseel® kit consists of the following components:
Tisseel®, a lyophilized, virus-inactivated Sealer, containing clottable protein, thereof: fibrinogen, Plasma fibronectin (CIG) and Factor XIII, and Plasminogen.
Aprotinin Solution (bovine)
Thrombin 4 (bovine)
Thrombin 500 (bovine)
Calcium Chloride solution
The Tisseel® kit contains a DUPLOJECT® Application System. The fibrin adhesive or the two-component sealant using Tisseel® Kit is combined in the following manner according to the Immuno AG product insert sheet:EXAMPLE 3
Chondrocytes were grown in minimal essential culture medium containing HAM F12 and 15 mM Hepes buffer and 5 to 7.5% autologous serum in a CO2 incubator at 37° C. and handled in a Class 100 laboratory at Verigen Europe A/S, Symbion Science Park, Copenhagen, Denmark. Other compositions of culture medium may be used for culturing the chondrocytes. The cells were trypsinized using trypsin EDTA for 5 to 10 minutes and counted using Trypan Blue viability staining in a Bürker-Türk chamber. The cell count was adjusted to 7.5×105 cells per ml. One NUNCLON™ plate was uncovered in the Class 100 laboratory.
The Surgicel® hemostatic barrier was cut to a suitable size fitting into the bottom of the well in the NUNCLON™ tissue culture tray. In this case a circle, of a size of approximately 4 cm (but could be of any possible size) and placed under aseptic conditions on the bottom in well in a NUNCLON™ Delta 6-well sterile disposable plate for cell research work (NUNC, InterMed, Roskilde, Denmark). The hemostatic barrier to be placed on the bottom of the well was pre-treated as described in Example 1. This treatment delays the absorption of the Surgicel significantly. This hemostatic barrier was then washed several times in distilled water and subsequently several times until non-reacted glutaraldehyde was washed out. A small amount of the cell culture medium containing serum was applied to be absorbed into the hemostatic barrier and at the same time keeping the hemostatic barrier wet at the bottom of the well.
Approximately 106 cells in 1 ml culture medium were placed directly on top of the hemostatic barrier, dispersed over the surface of the hemostatic barrier pre-treated with 0.4% glutaraldehyde as described above. The plate was then incubated in a CO2 incubator at 37° C. for 60 minutes. An amount of 2 to 5 ml of tissue culture medium containing 5 to 7.5% serum was carefully added to the well containing the cells avoiding splashing the cells by holding the pipette tip tangential to the side of the well when expelling the medium. It appeared that the pH of the medium was too low (pH ˜6.8). The pH was then adjusted to 7.4 to 7.5. The next day some chondrocytes had started to grow on the hemostatic barrier, arranged in clusters. Some of the cells had died due to the low pH exposure prior to the adjustment of the pH. The plate was incubated for 3 to 7 days with medium change at day 3.
At the end of the incubation period the medium was decanted and cold refrigerated 2.5% glutaraldehyde containing 0.1M sodium salt of dimethylarsinic acid, (also called sodium cacodylate, pH is adjusted with HCl to 7.4), was added as fixative for preparation of the cell and supporter (hemostatic barrier) for later preparation for electron microscopy.EXAMPLE 4
Chondrocytes were grown in minimal essential culture medium containing HAM F12 and 15 mM Hepes buffer and 5 to 7.5% autologous serum in a CO2 incubator at 37° C. and handled in a Class 100 laboratory at Verigen Europe A/S, Symbion Science Park, Copenhagen, Denmark. Other compositions of culture medium may be used for culturing the chondrocytes. The cells were trypsinized using trypsin EDTA for 5 to 10 minutes and counted using Trypan Blue viability staining in a Bürker-Türk chamber. The cell count was adjusted to 7.5×105 cells per ml. One NUNCLON™ plate was uncovered in the Class 100 laboratory.
The Surgicel® (for use as the hemostatic barrier) was treated with 0.6% glutaric aldehyde for one minute as described in Example 1, and washed with 0.9% sterile sodium chloride solution or, preferably, with a buffer such as a PBS buffer or the culture medium such as MEM/F12, because pH after the glutaric aldehyde treatment is 6.8 and should preferably be 7.0 to 7.5. The Tisseel® was applied on both side of the Surgicel® using the DUPLOJECT® system, thus coating both sides of the Surgicel®, the patch intended to be used, with fibrin adhesive. The glue is left to dry under aseptic condition for at least 3 to 5 minutes. The "coated" hemostatic barrier was placed on the bottom of the well in a NUNCLON™ Delta 6-well sterile disposable plate for cell research work. A small amount of tissue culture medium containing serum was applied to be absorbed into the hemostatic barrier. Approximately 106 cells in 1 ml tissue culture medium containing serum was placed directly on top of the Hemostat, dispersed over the surface of the hemostatic barrier. The plate was then incubated in a CO2 incubator at 37° C. for 60 minutes. An amount of 2 to 5 ml of tissue culture medium containing 5 to 7.5% serum was carefully added to the well containing the cells avoiding splashing the cells by holding the pipette tip tangential to the side of the well when expelling the medium. After 3 to 6 days, microscopic examination showed that the cells were adhering to and growing into the Surgicel® in a satisfactory way suggesting that Surgicel® did not show toxicity to the chondrocytes and that the chondrocytes grew in a satisfactory manner into the Surgicel®.
The plate was incubated for 3 to 7 days with medium change at day 3. At the end of the incubation period the medium was decanted and cold refrigerated 2.5% glutaraldehyde containing 0.1M sodium salt of dimethylarsinic acid, also called sodium cacodylate, pH is adjusted with HCl to 7.4, was added as fixative for preparation of the cell and supporter (hemostatic barrier) for later preparation for electron microscopy.EXAMPLE 5
Chondrocytes were grown in minimal essential culture medium containing HAM F12 and 15 mM Hepes buffer and 5 to 7.5% autologous serum in a CO2 incubator at 37° C. and handled in a Class 100 laboratory at Verigen Europe A/S, Symbion Science Park, Copenhagen, Denmark. The cells were trypsinized using trypsin EDTA for 5 to 10 minutes and counted using Trypan Blue viability staining in a Bürker-Türk chamber. The cell count was adjusted to 7.5×105 to 2×106 cells per ml. One NUNCLON™ plate was uncovered in the Class 100 laboratory.
It has been found that the Bio-Gide® can be used as a resorbable bilayer membrane which will be used as the patch or bandage covering the defective area of the joint into which the cultured chondrocytes are being transplanted as well as the hemostatic barrier. The Bio-Gide® is a pure collagen membrane obtained by standardized, controlled manufacturing processes (by E. D. Geistlich Söhne AG, CH-6110 Wolhusen). The collagen is extracted from veterinary certified pigs and is carefully purified to avoid antigenic reactions, and sterilized in double blisters by &ggr;-irradiation. The bilayer membrane has a porous surface and a dense surface. The membrane is made of collagen type I and type III without further cross-linking or chemical treatment. The collagen is resorbed within 24 weeks. The membrane retains its structural integrity even when wet and it can be fixed by sutures or nails. The membrane may also be "glued" using fibrin adhesive such as Tisseel® to the neighboring cartilage or tissue either instead of sutures or together with sutures.
The Bio-Gide® was un-covered in a class 100 laboratory and placed under aseptic conditions on the bottom of the wells in a NUNCLON™ Delta 6-well sterile disposable plate for cell research work,—either with the porous surface of the bilayer membrane facing up or with the dense surface facing up. Approximately 106 cells in 1 ml tissue culture medium containing serum was placed directly on top of the Bio-Gide®, dispersed either over the porous or the dense surface of the Bio-Gide®. The plate was then incubated in a CO2 incubator at 37° C. for 60 minutes. An amount of 2 to 5 ml of tissue culture medium containing 5 to 7.5% serum was carefully added to the well containing the cells avoiding splashing the cells by holding the pipette tip tangential to the side of the well when expelling the medium.
On day 2 after the chondrocytes were placed in the well containing the Bio-Gide® the cells were examined in a Nikon Inverted microscope. It was noticed that some chondrocytes had adhered to the edge of the Bio-Gide. It was of course not possible to be able to look through the Bio-Gide® itself using this microscope.
The plate was incubated for 3 to 7 days with medium change at day 3. At the end of the incubation period the medium was decanted and cold refrigerated 2.5% glutaraldehyde containing 0.1M sodium salt of dimethylarsinic acid, also called sodium cacodylate, pH is adjusted with HCl to 7.4, was added as fixative for preparation of the cell and the Bio-Gide® supporter with the cells either cultured on the porous surface or the dense surface. The Bio-Gide® patches were then sent for electron microscopy at Department of Pathology, Herlev Hospital, Denmark.
The electron microscopy showed that the chondrocytes cultured on the dense surface of the Bio-Gide® did not grow into the collagen structure of the Bio-Gide®, whereas the cells cultured on the porous surface did indeed grow into the collagen structure and furthermore, showed presence of proteoglycans and no signs of fibroblast structures. This result shows that when the collagen patch, as for instance a Bio-Gide® patch is sewn as a patch covering a cartilage defect the porous surface shall be facing down towards the defect in which the cultured chondrocytes are to be injected. They will then be able to penetrate the collagen and produce a smooth cartilage surface in line with the intact surface, and in this area a smooth layer of proteoglycans will be built up. Whereas, if the dense surface of the collagen is facing down into the defect the chondrocytes to be implanted will not integrate with the collagen, and the cells will not produce the same smooth surface as described above.EXAMPLE 6
Chondrocytes were grown in minimal essential culture medium containing HAM F12 and 15 mM Hepes buffer and 5 to 7.5% autologous serum in a CO2 incubator at 37° C. and handled in a Class 100 laboratory at Verigen Europe A/S, Symbion Science Park, Copenhagen, Denmark. The cells were trypsinized using trypsin EDTA for 5 to 10 minutes and counted using Trypan Blue viability staining in a Bürker-Türk chamber. The cell count was adjusted to 7.5×105 to 2×106 cells per ml. One NUNCLON™ plate was uncovered in the Class 100 laboratory.
The Bio-Gide® used as a resorbable bilayer membrane may also be used together with an organic glue such as Tisseel® with additional, significantly higher content of Aprotinin than normally found in Tisseel®, as described in the product insert. By increasing the content of Aprotinin to about 25,000 KIU/ml, the resorption of the material will be delayed by weeks instead of the normal span of days.
To test this feature in vitro, the Tisseel® is applied to the bottom of the well of the NUNCLON™ plate, and allowed to solidify incompletely. A collagen patch such as a Bio-Gide® is then applied over the Tisseel® and glued to the bottom of the well. This combination of Bio-Gide® and Tisseel® is designed to be a hemostatic barrier that will inhibit or prevent development or infiltration of blood vessels into the chondrocyte transplantation area. This hybrid collagen patch can now be used for both as a hemostatic barrier at the bottom of the lesion (most proximal to the surface to be repaired) but also as a support for cartilage formation because the distal surface can be the porous side of the collagen patch and thus encourage infiltration of chondrocytes and cartilage matrix. Thus this hybrid collagen patch can also be used to cover the top of the implant with the collagen porous surface directed down towards the implanted chondrocytes and the barrier forming the top. The hybrid collagen patch, with elevated Aprotinin component may also be used without any organic glue such as Tisseel® and placed within the defect directly, adhering by natural forces. Thus the collagen patch can be used both as the hemostatic barrier, and the cell-free covering of the repair/transplant site, with the porous surfaces of the patches oriented towards the transplanted chondrocytes/cartilage. Another variant would use a collagen patch which consists of type II collagen (ie. from Geistlich Sohne AG, CH-6110 Wolhusen).
Thus the instant invention provides for a hybrid collagen patch where said patch is a collagen matrix with elevated levels of aprotinin component, preferably about 25,000 KIU/ml, in association with an organic matrix glue, where the collagen component is similar to the Bio-Gide® resorbable bilayer material or Type II collagen, and the organic glue is similar to the Tisseel® material. In another embodiment, the hybrid collagen patch does not use any organic glue to adhere to the site of the repair.EXAMPLE 7
Because of the weakened structure of osteoarthritic cartilage, adherence of cultured autologous chondrocytes transplanted to a graft site in defective cartilage may be inhibited, thus creating a marginal zone (zone of demarcation) between the newly implanted cartilage/chondrocytes and the surrounding established cartilage. This marginal zone will be most pronounced if the graft site is prepared for the graft by creating straight, smooth walls cut in a linear fashion. The shearing and compression forces across such a marginal zone (as illustrated in FIG. 3A) will exert great force to dislodge the graft when the graft site is cut in a linear fashion. This marginal zone, and differential movement of materials along this zone will inhibit confluent healing between the grafted material and the surrounding material. In many cases the graft material is softer than the surrounding material, however, in some instances of osteoarthritis disease, the surrounding cartilage may in fact be softer than the implanted chondrocytes/cartilage.
Therefore, in order to solve this problem, the method of the invention teaches the use of surgical instruments to sculpt the walls of the graft site such that the walls are non-linear, and thus provide for undulated surfaces. It is also possible to shape the graft site such that the diameter of the site proximal to the bone surface is of a greater dimension then the opening distal to the bone, and at the surface of the cartilage. However, the preferred embodiment describes the sculpting of the walls of the graft site in an fashion similar to a threaded opening for receiving a bolt or screw (as illustrated in FIG. 3B), thus providing mechanical resistance to the compression and or ejection of the grafted material from the graft site which can be described as "male" and "female" threading.
The surgical instruments contemplated by the instant invention can be manufactured from metal and/or plastic suitable for making single-use disposable, or multi-use reusable surgical instruments. As cartilage is a relatively soft material it may be advantageous to manufacture hardened plastic cutting edges which will be able to sculpt cartilage without being able to damage bone. Such cutting instruments can be manufactured to incorporate openings for administration of fluid, suction removal of cutting debris and fluid, and fiber optic threads for illumination and visualization of the defect site. In certain embodiments of the instrument, the base of the instrument may have protruding point or pin-like structure which will assist in guiding and placing the instrument in the graft site. Of course such a pin would be designed to minimized damage to the underlying bone.
While the cutting surface of the instrument may be single toothed, or multi-toothed, or describe a screw-like pattern such as that in a metal tap used to generate threaded holes in metal parts, the characteristic required of the cutting instrument is that the resulting sculpted sides of the graft site is undulated, and non-linear. For example, in certain embodiments, the cutting edge of the instrument can be shaped similar to that shown in FIG. 4A, or as in FIG. 4B. The cutting edge maybe flat, or circular in that it wraps around the diameter of the cutting instrument. Many other shapes can be designed to accomplish the purpose of the method of the invention to create an interface which provides for mechanical resistance to differential reaction to compression and shearing forces on the transplanted material and the surrounding material.EXAMPLE 8
A four month old mixed Yorkshire breed pig was subjected to general anesthesia and placed on its back. The pig was washed and draped in a surgical suite at Harrington Arthritis Research Center, Phoenix, Ariz. The entire surgical procedure was performed aseptically. The left hind-leg and adjacent abdomen and inguinal area was cleaned with iodine. The knee joint was localized, and the patella localized. A medial incision was performed approximately 3 cm from the posterior part of the patella and the several subcutis, muscle layers and ligaments was cut approximately in order to get access to the medial femoral condyle. Using a circular cutter a lesion was prepared in the white cartilage on the medial part of the medial condyle, leaving a 0.5 to 1 cm margin to the edge of the cartilage covering the posterior-medial part of the condyle (left condyle, FIG. 6A). The 0.5 to 1 cm defect was placed in a caudal weight bearing part of the medial condyle. The entire surgical procedure was done without tourniquet on the left femur. The different layers and skin was sutured appropriately.
On day 3 the animal was again brought to the surgical suite and positioned as above on the operating table and given general anesthesia. The left hind leg, abdomen and inguinal region was ionized as described above. Sutures were cut and the area opened. It was noticed that a moderate hematoma was present in the knee joint. The blood clot was removed and the defect inspected. There was a blood clot in the defect which was removed. A sterile surgical instrument designed with a male thread cutting edge, with a size corresponding to, or slightly bigger than the circumference of the lesion was carefully screwed down into the defect. A BioGide® pad was cut to a size equal to the bottom of the defect. The first glue used, called Adhesive Protein (A-2707, Sigma Chemical, USA) was applied on the dense side of the trimmed hemostatic barrier pad, and the pad was placed dense side down into the bottom of the lesion, using it as a barrier as described above. It was found that this glue did not seem to dry very fast. The slight bleeding from the bottom of the defect stopped immediately. A second BioGide® was cut somewhat bigger in circumference than the lesion and was placed with dense side up (thus the porous side down towards the graft) as described above.
This non-cellular covering-pad was then sutured over the cavity, leaving one edge open, where the chondrocyte to be explanted could be injected. The surrounding part of the edge of the pad was covered with the second glue, Dow Corning Medical Adhesive B (Cat. #895-3, Dow Corning, USA). This second glue dried much faster and more efficiently than the first glue. It was found that during this particular procedure, the first glue had not dried sufficiently to hold the hemostatic barrier in place when suturing of the cap was attempted. Thus the main barrier formed on the proximal surface of the graft site was by the glue itself.
Using a 1 ml syringe and a 16 gauge needle, the chondrocyte cell suspension (about 0.6 ml) was drawn up into the barrel of the syringe. A 23 gauge short needle was switched for the 16 gauge needle, and the cell suspension was injected under the sutured covering-patch into the graft site (about 10×106 cells). The open edge of the cap was then glued prior to removal of the needle, and the needle carefully withdrawn. No leakage of cells was seen. The wound was sutured and as above, no tourniquet was used, no bleeding was observed. The final skin layers were sutured. No protrusion of the skin occurred after suturing, which indicates that there was no hematoma. Postoperative recovery was uneventful.
As expected, the grafted chondrocytes produced cartilage matrix sufficient to repair the defect made in the articular cartilage surface of the knee joint of the test pig. FIG. 6A is an MRI image of a pig knee showing the cartilage defect created in the knee (left condyle, the medial condyle), and FIG. 6B is an MRI image of the same pig knee three months after treatment showing repair of the defect.EXAMPLE 9
A kit comprising the components useful for practicing the method of the invention, will allow for the convenient practice of the method of the invention in a surgical setting. In a preferred embodiment, a kit of the invention will provide sterile components suitable for easy use in the surgical environment, and will provide a suitable hemostatic barrier, suitable covering patch, and if needed organic glue. A kit of the invention may also provide sterile, cell-free matrix material suitable for supporting autologous chondrocytes that are to be implanted into an articular joint surface defect. In one embodiment, a kit of the invention contains a Surgicel® hemostatic barrier and a Bio-Gide® covering patch with suitable coating of Tisseel® organic glue, where the Surgicel® and Bio-Gide® have been treated according to the teachings of the invention to increase the time till resorption. In instances where Tisseel® is pre-coated, in one embodiment the Tisseel® is supplemented with additional aprotinin to increase time till resorption.
In another preferred embodiment, the hemostatic barrier and covering-patch are both a semi-permeable collagen matrix which is treated to extend the time till resorption of the material. It is also possible to provide Tisseel® glue in enhanced form as a separate component to be applied as needed because of the inherent variability and unique circumstances every repair/transplantation procedure will encounter.
A further embodiment of the kit will include a surgical instrument as described in Example 7 above.
It will be appreciated by persons skilled in the art that numerous variations and/or modifications may be made to the invention shown in the specific embodiments without departing form the spirit and scope of the invention as broadly described.
1. A method for the effective treatment of articulating joint surface cartilage by the transplantation of chondrocytes in a suitable matrix, to a surface to be treated, the method comprising the steps:
- (a) sculpting the surface to be treated;
- (b) placing chondrocytes in a suitable matrix upon the surface to be treated; and
- (c) covering the surface to be treated with a covering-patch.
2. The method according to claim 1, wherein the matrix is cell-free.
3. The method according to claim 1, wherein the covering-patch is a semi-permeable collagen matrix with a porous surface.
4. The method according to claim 3, wherein the porous surface of the covering-patch is directed toward the surface to be treated.
5. The method according to claim 1, wherein the covering-patch is a semi-permeable collagen matrix which is substantially free of intact cells.
6. The method according to claim 1, wherein the covering-patch is resorbable.
7. The method according to claim 1, wherein the covering-patch is partially attached to the surface to be treated prior to placing of the chondrocytes in a suitable matrix on the surface to be treated in said step (b).
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Type: Grant
Filed: May 26, 1999
Date of Patent: Sep 4, 2001
Assignee: Verigen Transplantation Services Internt'l (Leverkusen)
Inventors: Henrik Vibe-Hansen (Lyngby), Charlotte Lundsgaard (Clampenborg), Kurt B. Osther (Scottsdale, AZ), Abmed Idouraine (Chandler, AZ)
Primary Examiner: Cary E. O'Connor
Attorney, Agent or Law Firm: Ratner & Prestia
Application Number: 09/320,246
International Classification: A61B/1708;

subset: commoncrawl


text:
Don't Go North on the Blue Bridge! Avoid Cable Bridge If You Can
Andrew Kirk[/caption]Northbound lanes on the Blue Bridge are closed this morning for repairs and will be through early Nov. 5. Traffic is being directed onto Columbia Drive and then the Cable Bridge. That means in the mornings Columbia Drive and Gum Street are PACKED with cars and semi-trucks trying to get to Pasco. If you can avoid it, AVOID IT! People in west Kennewick and all Richland residents are advised to take 182 to Pasco. Here are more pics from this morning:
Andrew Kirk[/caption]
Andrew Kirk[/caption]


text:
'Star Wars Rebels' Poster and Official Plot March Out: Beware the Evil Empire!
'Star Wars Rebels' will likely keep its cards close to the chest, even through tomorrow's New York Comic-Con panel, but that won't stop the new mid-trilogy animated drama from recruiting early. A new propaganda poster and the official plot description for the 'Star Wars' TV series has emerged to follow this past week's teaser, but for the dark side or the light? Join the rebels and fight the Empire with our latest look at the series!
Per Entertainment Weekly, the new 'Star Wars Rebels' poster borrows a bit from old Soviet propaganda posters to highlight Empire recruitment in a stylized depiction of stormtroopers. The poster also includes a bit of Aurabesh writing, which if the comments are to be believed, ominously spell out, "Be A Part of Tomorrow - Today."
According to the official 'Star Wars Rebels' plot from Lucasfilm:
'Star Wars Rebels' continues the epic tradition of the legendary 'Star Wars' saga with all-new exciting, action-packed adventures. It is a dark time in the galaxy, as the evil Galactic Empire tightens its grip of power from world to world. As the series begins, Imperial forces have occupied a remote planet, ruling with an iron fist and ruining the lives of its people. But there are a select few who are brave enough to stand up against the endless Stormtroopers and TIE fighters of the Empire: the clever and motley crew of the starship "Ghost." Together, this ragtag group will face threatening new villains, have thrilling adventures, and become heroes.
We've already seen concept art of the mentioned Millennium Falcon-esque "Ghost," which will act as a central location, but no voice cast has been made official. Though, we recently heard Freddie Prinze Jr., 'The Butler' star David Oyelowo, Vanessa Marshall, Taylor Gray and Steven Jay Blum were on the short list.
Picking up where the now-canceled 'Star Wars: The Clone Wars' left off, 'Star Wars Rebels' will feature on the creative side screenwriter and producer Simon Kinberg ('X-Men: First Class,' 'Elysium'), who's writing the first episode, as well as 'Clone Wars' supervising director Dave Filoni and 'Young Justice' producer Greg Weisman.
We'll be on hand this weekend to learn more about 'Star Wars Rebels' at NYCC. In the meantime, check out the new poster below, and tell us what you want to see from the new 'Star Wars' in the comments!


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I've been on a miso kick. It all started when a friend gave me a container of very special, well-aged, Korean miso. It was familiar and comfortable, like sitting in your father's TV-watching chair as an adult. Not that I hadn't been using miso because I had been using it, but primarily as a marinade for fish and meat, and occasionally as a dressing on green beans or asparagus.
The first time I had miso was probably the way almost everyone did, as miso soup. I was 20 and at Ottawa's only Japanese restaurant with my Japanese boss. I remember we had sashimi, and there was miso soup to start. Tiny cubes of tofu bobbed in the bowl and sliced green onion floated on the surface. I was fascinated by the way the more solid parts of the miso formed a "cloud," smack in the centre of the bowl. I thought it was so beautiful, and the taste was otherworldly, like nothing I had experienced before. An excited voice in my head kept repeating "Sashimi! Miso soup! Sashimi! Miso soup!" I knew I was being indoctrinated into something exciting, and it was something that would become a lifelong passion: Japanese food.
A year later, I found myself submerged in the macrobiotic food movement. Based upon the principles of yin and yang, the balancing point was organic, short-grain brown rice. On either side of that balancing point were yin foods, yang foods, and foods to be avoided. The avoids were all members of the nightshade family — tomatoes, potatoes, eggplant — and not to be out-and-out avoided, yet barely consumed, animal protein. Local, in-season foods were good, imported, out-of-season foods were not good. Fruit was yin, and sugar was poison. Toothpaste was burnt eggplant mixed with sea salt, very yang. There were cold pressed oils, and root vegetables. And there was miso, umeboshi plums, tofu and tamari. The miso aspect I found very fascinating, mostly due to a book that came out that year called The Book of Miso. The authors, Bill Shurtleff and Akiko Aoyagi, had built a definitive tome on miso that has not been matched since. Everything from the history, the manufacturing, and on to making your own miso was in there. To be frank, the recipes were a little odd, as if the authors were so eager to get the word out that they had to cram miso into almost everything, though there were a number of traditional recipes too. The one recipe I glommed on to was miso mixed with tahini into a pate-like consistency. I relished it spread on rice cakes as a snack. The variety of miso that we have today was not available to me then. All I could find was a dark-brown type from a local natural foods store. It didn't matter because it was tasty and I knew from reading the book that miso was good for me.
Now, with my interest in fermented foods, curing meats and vegetables, Korean cuisine and a new love for naturally occurring umami, I'm back to exploring miso again. Tucked into the basement fridge is a tray of tofu misozuke, tofu blanketed in miso that in a few months will be transformed into a creamy "cheese." A slab of pork belly curing in miso will be ready in a few days and will be eaten with kimchee. There's a little miso-tofu mayonnaise, a faux mayo, but good nonetheless. And for experimentation purposes, there are five different types of miso for me to play with.
I may even try my hand at making miso one day, though I generally like to leave that type of thing to the experts. And is there a container of the miso tahini spread from years past? You betcha!
Miso Sablefish with Apple Slaw
Miso marinated sablefish has become quite popular in recent years and with good reason. For marinating time, one day is good, two is better, and three is fantastic.
In the top of a double boiler combine the miso, sugar, sake and mirin. Cook for 30 mins, stirring frequently, until the mixture is shiny and thick. Remove from the heat and let cool. Evenly coat the sablefish with the miso mixture and place into a container that fits the sablefish snugly, or into a Ziploc bag. Cover and refrigerate overnight, or up to three days.
Combine the mayonnaise with the mustard, miso and lemon peel. Cover and refrigerate until needed.
When you are ready to serve the fish, combine the apple, cabbage, green onion, pine nuts and sesame seeds. Add the mayonnaise. Mix well and refrigerate.
Heat the oven to 425°F (220°C). Remove the sablefish from the marinade, letting most of it drip off the fish. Place on a rimmed, foil covered baking sheet. Bake for 15 minutes. Remove the pan from the oven and look at the fillets. You'll notice the bones protruding. Try to pull one out. If it comes out easily, the fish is done. If not, return to the oven for 5 minutes longer. In either case, return to the oven and broil 8-inches (20 cm) from the heat until the fish browns deeply around the edges. Serve immediately with the slaw.
Makes 4 servings
Miso Roasted Cauliflower
Four ingredients with great results. The same mixture can be used for roasting Brussels sprouts, carrots, parsnips, and rutabaga.
Heat the oven to 425°F (220°C). Trim the cauliflower and cut it into 8 wedges. Stir the miso to loosen it up, stir in the mayonnaise and mustard until smooth. Coat all the surfaces of the cauliflower with the mixture — it's OK if it's patchy — and place on a rimmed, foil covered baking sheet. Bake for 10 minutes then turn the wedges over and bake for 10-15 minutes longer until the cauliflower is edged with brown. Serve warm or at room temperature.
Makes 4-6 servings
Avocado and Tomato Salad with Miso Carrot Dressing
The tomato and avocados are merely a starting point for using this delicious dressing. It goes with sturdy vegetables and greens such as cucumbers, beets, Brussels sprouts, asparagus or kale, and can be used as a dip or sauce. You'll find a myriad of ways to use it.
Place the miso, vinegar, oils and water into a blender. Buzz until smooth. Add the remaining ingredients and buzz again until smooth. The dressing is best made a few hours in advance to let the flavours mingle and emerge. Keeps for 1 week, covered and refrigerated
Arrange the avocados and tomatoes on a platter or individual plates. Drizzle with the dressing and sprinkle with the shallots. You may have more dressing than you need, depending on your taste. Serve immediately.
Makes 4-6 servings
Miso Caramel
If you like salted caramel, you will love this. The miso adds a great depth of flavour to the sauce. Use it on ice cream, pound cake, sliced apples or bananas, or eat it straight from a spoon.
Before starting this recipe, find a sieve that will fit snugly into the pot that you are using to make the caramel. When you pour the cream into the caramelized sugar, it will splutter furiously and the sieve will prevent the hot liquid from splashing everywhere. Wear oven mitts and stand back from the pot when pouring the cream.
Combine the sugar and water in a heavy, medium pot. Bring mixture to a boil over medium high heat without stirring. Use a wet pastry brush to wash down any sugar crystals that form on the sides of the pan. Swirl the pan occasionally to combine the caramelizing sugar. When the sugar becomes a deep golden brown and wisps of smoke just start to form, quickly remove the pot from the heat, place the sieve over the pot and pour the whipping cream through the sieve. When the spluttering stops, return the pot to the heat and stir to dissolve any masses of caramelized sugar. Remove from the heat and let cool. Whisk in the miso and strain through a sieve. Store covered and refrigerated for up to 2 weeks.
To return it to a warm, pourable sauce, reheat gently over low heat, adding a bit of water if the caramel is too thick.
Makes approximately 1 1/2 cups (375 mL)


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Fifth Harmony Demand Respect in New Video 'BO$$'by Mandi Salerno | PopCrush
A new era of Fifth Harmony is being ushered in with their video for 'BO$$.' Are you ready, Harmonizers?
Normani, Camila, Lauren, Dinah and Ally are serving up serious attitude in the video, dressed in sexy white jackets and shorties as they demand, "Yo ass betta show me some respect." Who can argue with that?
In each scene — sashaying down a catwalk, provocatively dancing over metal chairs, arm wrestling beefy dudes — they own the situation and prove that these aren't the same girls we saw as recently as their video for 'Me & My Girls.' Gone is the cutesy vibe, replaced with sexy confidence.
The product placement is sometimes a little distracting in the video, but the girls strutting their stuff to the catchy beat will draw you back in immediately. The snaps and heavy percussion are so powerful you'll feel them pounding in your pulse as 5H win you over with their hot moves and dynamic voices. There's no doubt who the bosses are in this video!
Check out Fifth Harmony's 'BO$$' up above.


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Just guessing, but this is probably exactly how Michael Phelps acted during that South Carolina frat party. (Examines fingers) ... "Is this real life?"
This kid was returning from the dentist, where, it appears, he was given a generous helping of happy gas. Remember that feeling, my young friend; lest your future be filled with restroom stall door collisions.
Greatest part: How he just conks out at the end. I wonder if they drove Phelps home in a car seat?
Little Boy Is Faded After The Dentist [Phil Knows Best]


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Amazon Music tracked the top country songs in each state based on share of streams for one month this summer—and there are plenty of interesting takeaways.
Some states have a lot of pride in their native sons:
- The top song in Illinois is "Illinois" by Illinois native Brett Eldredge.
- North Carolina's top song is "Carolina Can" by N.C native Chase Rice.
- The king is Texas is George Strait's "I Got a Car."
- Arizona likes native son Dierks Bentley's "I'll Be the Moon."
- Alabamians prefer Alabama's "Dixieland Delight."
- Kentucky's top song is native son Chris Stapleton's "Nobody to Blame"
Some states like to drink:
- Florida's top song is "It's Five O'Clock Somewhere" by Alan Jackson and Jimmy Buffet.
- Georgia's top song is "A Little Dive Bar in Dahlonega" by Ashley McBryde.
- Idaho's top song is "Drinking Class" by Lee Brice.
- Nebraska's top song is "Drink in My Hand" by Eric Church.
- New Mexico's top song is "Sangria" by Blake Shelton.
- Wisconsin's top song is "Dozen Roses and a Six Pack" by Cole Swindell.
Some artists are popular in multiple states:
- Jason Aldean had the top song in Colorado, Delaware and Indiana.
- Carrie Underwood had the top song in Maine, New Jersey and Utah.
- Keith Urban had the top song in California, New Jersey and Oregon.
- Miranda Lambert had the top song in Alaska, Maine and Oklahoma.
Check out the full list below, courtesy of Amazon Music.
photo by Tammie Arroyo, AFF-USA.com


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If this is true, we have a LOT of questions.
(Thanks to OriginalEnigma)
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If this is true, we have a LOT of questions.
(Thanks to OriginalEnigma)
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Someone please tell me where this idiot is so I can SMACK him silly???
News you can use!
If this is true, then a lot us have doing doing something wrong for a long time.
Notice how they blamed the woman's lack of enjoyment on the woman. It probably says more than a little about their technique and the size of their Walters.
So, the reason women don't enjoy sex is because they have no vagina? Or is it possible they don't like having a bag over their head while having sex and kept in a closet the rest of the time?
I've often raised them up to mythical status... (esp. when I was in my early yrs..)
um, eye think its a spoof.
ha, ha.
Uh, sure, Abdul Abdul, you're right of course.
"Prince Abdul Abdul Abdul" ri-i-i-i-ght.
Oh, excuse me. Shouldn't have had that explosive Phillie Cheesedog at the ballpark.
Siouxie, I think they're ALREADY silly. These are the guys with the pointed hats that wear dresses, after all. I'd say smack 'em stupid, but they've gone beyond that, as well.
A spoof?
Does that mean the story on the second page about Palin being a pit-bull/human hybrid is false also?
Well then, whose monologue have I been listening to?
I knew she was faking it.
It's not a real news story.
*snork* @ Cat R !!
**was going to make a snide remark about an echo, but I like you, Cat !**
This is obviously a spoof. However, some parts of that country have had a tradition of female genital mutilation, which excises the clitoris. Those women probably don't get much pleasure out of sex.
Someone has their turban wrapped a bit too tight.
Laugh. Out. Loud. Although, I am sure women over there do have a psychological aversion to sex for blatantly obvious reasons to anyone who does not live in that cuntry.
Okay, that site is hilarious!
*tosses a screaming "o" up to Margaritaville*
So not only do women fake it, so does the spoof-writer.
Maybe they are just using their own partners for the study.
Looks like South Africa's version of The Onion. Time to go throw some impala steaks on the brai.
My husband is going to be upset.
Unlike the Peoples'News or The New York Times, to get on the mailing list you MUST state your race. Could that be an S.A. thing? And there is no check-off for Human.
My error. Not S.A. but Za. (Zambia, Zaire, Zimbabwae? Help me out folks. Or not.)
This is obviously a spoof, but our Saudi "friends" wouldn't laugh, but then again, when DO they laugh?
"Clitoris was a common name for girl-children in the mid-Victorian era," said Prof. Saud, adding that it sounded quaint and "faintly reminiscent of flowers."
My grandmother's family had some pretty weird names, but Clitoris was not one of them...
Sadly, I now know my wife has GOT to be Muslim....
Abdul Abdul Abdul, also known as Abdul the Third.
I hope I'm not the only one who realized this is a fake news website? Their lead story (http://www.hayibo.com) is about godzilla.
People are spreading this around the web as "real news," though, which is troubling...
Yes, Cassa, you're the only one who gets the joke. Are you saying the Godzilla isn't real either?? Waking up tommorrow is going to be a relief for my kids. I've used Godzilla as a threat for years.
Shell, I LOVED that everyone just ignored the obvious and went with it. Sometimes you just gotta go for the humor and enjoy !
DELORIS!!!!!!! Kinda flowery, dontchya think?
BWAAHAHAHAHAHA!!!!! Omigod! BWAHHAHAHAHAHA!!! *tearing eyes* You gotta be kiddi...BWAHAHAHAA!!
**takes umbrage**
Ahem. My sainted Mother's name is Dolores !
Just kidding, Layzee - "Dolores" in spanish means "pain", sorta. She always goes by her nickname, Lola ...
The .za code stands for Zuid Afrika, which is Afrikaans for South Africa
The comments to this entry are closed.-


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I'm new, n' stuff! ^_^
**Ocy looks around.**
Dun worry about the emptyness. I've seen it before, in a different forum. I was one of the first posters, and guess what? That forum now has an occult following of over 2000 people..
Anyhow, Umm... I'm Ocy ^_^ Full name is Ocyrrhoe Trazere(RP name). Yep.
Ocy sets up residence, sittin' on top of a table.
[Edit] Silly, evil UBB..[/edit]
I believe in Jesus Tape, Savior of all things broken!
"I lay in be last night, staring up at the stars.. And I wondered to myself, "WHERE THE HELL IS THE CEILING!?!?"
[This message has been edited by -Ocy Trazere- (edited 12-19-2001).]


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Dental Care And Total Health
[You And Your Health]
Practicing good oral care helps you maintain healthy teeth and gums.
It also helps prevent stains from smoking, coffee, and soda. A strong set of teeth helps you chew food, and maintains the structure of your face, including your smile. Regular dental care is associated with good overall health. For example, caring for your teeth and gums can help prevent heart infections.
Diabetics, who are twice as likely to develop gum disease, may have a harder time regulating their blood sugar levels if bacteria from gum disease enter their bloodstream. Also, according to the American Academy of Periodontology, pregnant women with gum disease are seven times more likely to have a low-birth weight baby.
By following the tips below, you can enjoy a lifetime of healthy teeth, avoid painful dental treatments, and keep your body healthy.
• Adults and children should visit the dentist at least twice a year for routine cleaning and exams. Children should begin dental visits at 12 months of age or six months after their first tooth comes in – whichever comes first.
• Brush your teeth at least twice a day with toothpaste containing fluoride.
• Brush your teeth in a circular motion for at least two minutes, making sure to reach teeth in the back.
• Be sure to brush your gums and tongue.
• Change your toothbrush every three months.
• Floss daily.
• Eat a healthy diet rich in vegetables, fruit, whole grains, and dairy products. Limit or avoid sugary foods. But if you do eat sugar, brush your teeth right afterwards, if possible.
• Quit smoking – Smokers have a higher chance of developing gum disease and other health conditions, such as cancer and heart disease.
If you need a dentist and are enrolled in Affinity's dental plan, please visit Aetna's web site at aetna.com for a list of providers in your area. If you are not enrolled with Aetna, you may visit the American Dental Association's (ADA) site at http://www.ada.org/public/disclaimer.asp to find an ADA-affiliated dentist.
Dr. Citerman works at Affinity Health Plan, an independent, not-for-profit organization offering quality health care coverage to underserved New Yorkers. For more information visit www.affinityplan.org or call (866) 247-5678.
For questions about dental or general health, e-mail Dr. Citerman at email@example.com.
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I think you have "Filtered HTML" turned on here. If you switch to "Full HTML," it will probably...


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Bucs slide to 0-5 to start 2012 campaign
DES MOINES, Iowa. (Feb. 9, 2012) – The No. 74 ETSU men's tennis team continued its early-season struggles, falling to No. 73 Drake 6-1 on Thursday inside the Roger Knapp Tennis Center.
"It's so frustrating to have lost our way and belief to win," Director of ETSU Tennis Yaser Zaatini said. "We play well in spurts but can't pull out a complete match, and I take full responsibility for our incapacity to get it together.
"We're lacking fire, desire, motivation, and we are scared to lay in on the line. I know these men have a lot to give, and I know these Bucs are winners and I'm convinced that we'll wake up before too long."
ETSU (0-5) began the afternoon by falling in all three doubles matches. Junior Juan Ramirez (Asuncion, Paraguay) and sophomore Jorge Varon (Valencia Venezuela) were the first to fall victim to Drake (5-1), losing 8-2 to Jean Erasmus and Robin Goodman. Senior Sander GIlle (Kuringen, Belgium) and junior Jesus Bandres (Caracas, Venezuela), along with senior Robin Akser (Ornskoldsvik, Sweden) and senior Jeremy Bonnevalle (Keerbergen, Belgium) soon followed suit, with each pairing falling 8-5 to give the Bulldogs the doubles sweep and a 1-0 match lead.
Bonnevalle and Ramirez found no better luck, each falling in straight sets with Goodman defeating Bonnevalle 6-3, 6-4 at No. 5 singles, and James McKie defeating Ramirez 6.2, 6-3 at the No. 3 spot.
With a 3-0 lead in the match, Drake needed just one more point to clinch its fifth win of the season, but was halted when Akser put the Bucs on the scoreboard by taking out Alen Salibasic 6-4, 4-6, 6-2. The win improved Akser to 5-5 overall, 1-1 in dual play, and 1-0 at No. 4 singles.
The comeback hopes didn't last too long for ETSU, as Erasmus came back from a set down to defeat Bandres 6-7 (5-7), 6-4, 6-2 at No. 2 singles. The win locked up the match for Drake, and gave the Bulldogs their first win over a ranked team – while also being ranked – since 2008.
The remaining two matches saw Gille fall to No. 51 Anis Ghorbel in a highly competitive 6-7 (4-7), 6-4, 7-6 (7-5) match, and Varon drop a 4-6, 6-1, 6-1 decision to Ben Mullis.
The Bucs move on to play Denver on Saturday in Lincoln, Neb. at 2 p.m.
For more information on ETSU tennis, please visit ETSUBucs.com and click on the tennis links.


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