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Methods of treating and forming biocompatible microcapsules that contain living cells are provided, to improve the function of the microcapsules. In particular, methods of treating islet cells or microcapsules containing islet cells are provided. Culture of isolated islet cells prior to encapsulation, culture of encapsulated cells, and cryopreservation of islet cells prior to encapsulation, are described. Methods for harvesting viable islets that incorporates an anti-oxidant in the digestion medium are also disclosed.
1. A microencapsulated islet cell product comprising microcapsules containing isolated living pancreatic islet cells therein, said microcencapsulated islet cells exhibiting a weight gain of not more than 10 percent by weight over a period of one month in physiological saline solution at 37 degrees Celsius and exhibiting at least 150 percent basal insulin secretion in response to 16.7 milliMolar glucose challenge in Krebs-Ringer physiological solution at pH 7.4 after said period of one month.
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11. A microencapsulated islet cell product comprising microcapsules containing isolated living pancreatic islet cells therein, said microcencapsulated islet cells exhibiting a weight gain of not more than 10 percent by weight over a period of one month in physiological saline solution at 37 degrees Celsius and exhibiting at least 150 percent basal insulin secretion in response to 16.7 milliMolar glucose challenge in Krebs-Ringer physiological solution at pH 7.4 after said period of one month;
wherein said microcapsule comprises a polysaccharide gum surrounded by a semipermeable membrane;
and wherein said microcapsule has a diameter of from about 300 μm to about 700 μm.
12. A microencapsulated islet cell product according to
This application is a divisional of U.S. Application Ser. No. 09/453,348 filed Dec. 1, 1999, now U.S. Pat. No. 6,365,385, which is a continuation-in-part of commonly owned, application Ser. No. 09/237,407, filed Mar. 22, 1999, now U.S. Pat No. 6,303,355, the disclosures of which are incorporated by reference herein in their entirety.
The present invention relates to methods of treating isolated pancreatic cells and microencapsulated pancreatic cells, in order to prepare the cells for transplantation. Media containing an antibiotic, an anti-oxidant, an anti-cytokine, or an anti-endotoxin, or combinations thereof, are utilized.
Glycemic control in diabetes has been shown to delay the onset of, and slow the progression of, associated pathological complications. However, achieving adequate glycemic control using insulin therapy can be difficult. One alternative to insulin therapy is the transplantation of functioning pancreatic islet cells to diabetic subjects, to provide biological insulin replacement.
Approximately one percent of the volume of the human pancreas is made up of islets of Langerhans (hereinafter "islets"), which are scattered throughout the exocrine pancreas. Each islet comprises insulin producing beta cells as well as glucagon containing alpha cells, somatostatin secreting delta cells, and pancreatic polypeptide containing cells (PP-cells). The majority of islet cells are insulin-producing beta cells.
However, transplanted or grafted islet cells encounter immunological rejection, which can limit the clinical usefulness of his method. (Brunicardi and Mullen, Pancreas 9:281 (1994); Bartlett et al., Transplant. Proc. 26:737 (1994)). To combat rejection, immunosuppressive drugs may be used, but such immunosuppressive therapy impairs the body's immunological defenses and carries significant side effects and risks in itself. Approaches to containing and protecting transplanted islet cells have been proposed, including the use of extravascular diffusion chambers, intravascular diffusion chambers, intravascular ultrafiltration chambers, macroencapsulation and microencapsulation. The goal of islet transplantation is to achieve normoglycemia in the treated subject for some extended period of time.
Microencapsulation of islet cells has been proposed to reduce or avoid immunological rejection of transplanted islet cells. Lim and Sun, Science 210:908 (1980). The cells are encapsulated in a membrane that is permeable to cell substrates and cell secretions, but essentially impermeable to bacteria, lymphocytes, and large immunological proteins. The method of microencapsulation described by Lim and Sun involves forming gelled alginate droplets around isolated islet cells, and then adding coats of poly-L-lysine and additional alginate. The inner gelled core of the microcapsule is then liquefied by chelation. However, chelation of the core affects the structural support of the capsules and may adversely affect durability. The success of microencapsulated islet cell transplantation in treating diabetes depends on the ability of the microcapsules to provide sufficient amounts of insulin in response to glucose stimulation, over an extended period of time, to achieve adequate glycemic control.
Methods of treating isolated pancreatic cells, or of treating microencapsulated pancreatic cells, to enhance glucose-stimulated insulin production by the microcapsules and to provide durable microcapsules capable of glucose-stimulated insulin production, are therefore desirable.
A first aspect of the present invention is a method of treating isolated living cells, by first culturing the cells in a medium containing at least one of (or a combination of ): an antioxidant, an anti-cytokine, an anti-endotoxin, or an antibiotic. The cells are then microencapsulated in a biocompatible microcapsule that contains a hydrogel core and a semipermeable outer membrane, to provide a microcapsule containing living cells therein.
A further aspect of the present invention is a method of treating isolated living cells, by first cryopreserving the cells in a cryopreservation medium containing at least one of (or a combination of): an antioxidant, an anti-cytokine, an anti-endotoxin, or an antibiotic; then thawing the cells and encapsulating the cells in a biocompatible microcapsule having a hydrogel core and a semipermeable outer membrane.
A further aspect of the present invention is a method of treating biocompatible microcapsules containing living cells, where the microcapsule contains a hydrogel core and a semipermeable outer membrane. The microcapsules are cultured in a medium containing at least one of (or a combination of): an antioxidant, an anti-cytokine, an anti-endotoxin, or an antibiotic.
A further aspect of the present invention is a method of preparing microencapsulated cells by first culturing the cells in a cell culture medium containing at least one of (or a combination of): antioxidants, anti-cytokines, anti-endotoxins, and antibiotics. The cells are then encapsulated in a biocompatible microcapsule having a hydrogel core and a semipermeable outer membrane, where the living cells are present in the core. The microcapsules are then cultured in a medium containing at least one of (or a combination of): an antioxidant, an anti-cytokine, an anti-endotoxin, and an antibiotic.
A further aspect of the present invention is a method of preparing microencapsulated cells that includes a step of incubating the microencapsulated cells with a physiologically acceptable salt such as sodium sulfate or the like in order to produce a more durable, and therefore useful, biocompatible microcapsule.
A further aspect of the present invention is a method of isolating pancreatic islet cells in which an antioxidant is included in the digestion medium that is used to free the islet cells from pancreatic tissue.
A further aspect of the present invention is microencapsulated cell products, which may be produced by a process as described above.
FIG. 1 graphs glucose-stimulated insulin secretion in control islets (n=4). Following a one hour period of preperifusion of unencapsulated islets with Krebs-Ringer Ringer-bicarbonate (KRB) containing 3.3 mM (basal) glucose, basal effluent perifusate was collected over 20 minutes. The glucose concentration in the perifusate was then raised to 16.7 mM and after 30 minutes of perifusion with sample collection, the glucose concentration was reduced to basal during another 20 minutes of perifusion.
FIG. 2 graphs the effect of chelation on the microcapsular core. Using the protocol described for FIG. 1, above, islets enclosed in microcapsules with a liquefied (chelated) core were tested for glucose-stimulated insulin production.
FIG. 3 graphs glucose stimulation of islets in unchelated microcapsules. Using the protocol described for FIG. 1, above, islets enclosed in microcapsules with a gelled (unchelated) core were tested for glucose-stimulated insulin production.
FIG. 4 graphs the effect of 24 hours of culture on gelled (unchelated) microcapsule function. Following microencapsulation of islets without chelation, the microcapsules were cultured in RPMI 1640 medium for 24 hours prior to testing the response of the enclosed islets to glucose stimulation.
FIG. 5 shows the capsule weight in grams over time in days of control microcapsules and microcapsules treated with sodium sulfate to enhance the durability thereof. The lack of weigh gain over time for the treated microcapsules indicates that stability of such microcapsules.
FIG. 6 is a comparison of the percent basal insulin secretion of control and sodium sulfate treated islet capsules. Note that sodium sulfate treated cells exhibited substantially the same responsiveness as the control cells, indicating that the sodium sulfate treatment step used to enhance the durability or stability of the microcapsules was not unduly deleterious to the cells encapsulated therein.
The present inventors have determined that biocompatible microcapsules that contain living cells, such as pancreatic islet cells, benefit from a period of culture prior to use. Such culture enhances the ability of microcapsules containing islet cells to produce insulin in response to glucose stimulation. Culture of cell-containing microcapsules in a medium containing any of, or a combination of, an antibiotic, an anti-oxidant, an anti-cytokine, and an anti-endotoxin, is preferred.
The present inventors have further determined that cryopreservation of isolated cells (such as pancreatic islet cells) prior to microencapsulation does not adversely affect the function of the cells when subsequently encapsulated. Cryopreservation in a medium containing any of (or a combination of) an antibiotic, an anti-oxidant, an anti-cytokine, and an anti-endotoxin is preferred.
The present inventors have further determined that pre-culturing isolated cells prior to microencapsulation is beneficial. Culture of isolated islet cells prior to microencapsulation improves the glucose-stimulated insulin response of the encapsulated islets. Culture in a medium containing any of (or a combination of) an antibiotic, an anti-oxidant, an anti-cytokine, and an anti-endotoxin is preferred.
The above methods of treating cells and microcapsules may be combined, for example, by culturing isolated cells prior to microencapsulation and then culturing the resulting microcapsules.
Isolating Islets
Methods of isolating pancreatic islet cells are known in the art. Field et al., Transplantation 61:1554 (1996); Linetsky et al., Diabetes 46:1120 (1997). Fresh pancreatic tissue can be divided by mincing, teasing, comminution and/or collagenase digestion. The islets are then isolated from contaminating cells and materials by washing, filtering, centrifuging or picking procedures. Methods and apparatus for isolating and purifying islet cells are described in U.S. Pat. Nos. 5,447,863, 5,322,790, 5,273,904, and 4,868,121. The isolated pancreatic cells may optionally be cultured prior to microencapsulation, using any suitable method of culturing islet cells as is known in the art. See e.g., U.S. Pat. No. 5,821,121. Isolated cells may be cultured in a medium under conditions that helps to eliminate antigenic components (Transplant. Proc.14:714-23 (1982)).
In general, a method of isolating pancreatic islet cells comprises (a) digesting pancreatic tissue with a digestion medium, the digestion medium containing an antioxidant, with the digesting step carried out for a time sufficient to produce free pancreatic islet cells suitable for subsequent harvesting; and then (b) collecting the free pancreatic islet cells to produce isolated pancreatic islet cells. The antioxidant is included in the digestion medium in an amount sufficient to inhibit reoxygenation injury of the isolated pancreatic islet cells (e.g., from about 0.05, 0.1, or 0.5 milliMolar to about 5, 10 or 20 milliMolar or more) The antioxidant is preferably one that is not itself digested in the digestion medium, such as a vitamin or organic chemical antioxidants. Examples of suitable antioxidants include, but are not limited to, vitamin C, vitamin E, vitamin K, lipoic acid, lazaroids, and butylated hydroxyanisole. Once collected, the isolated pancreatic islet cells may be cultured and microencapsulated as further described herein.
A preferred method for isolating viable islet cells, such as porcine islet cells, is as follows. Adult pigs (approximately 25-20 kg) are anesthetized, followed by intra-arterial infusion of ice-cold UW solution and complete pancreatectomy. The pancreatic duct is cannulated for infusion of digestion medium comprising an antioxidant. An exemplary digestion medium is Hanks' balanced salt solution containing collagenase type P (1.5 mg/ml), DNase 1 (10,000 units) and TROLOX® brand water soluble vitamin E antioxidant (1 mM). After 20 minutes incubation on ice, the pancreas is incubated at 37° C. for 20 minutes before being hand-shaken for one minute. Digested tissue is filtered, and islet clusters are separated using OPTIPREP® gradient, washed in Hanks' solution and identified by dithizone staining. This method utilizes an anti-oxidant in the digestion medium and hand-shaking, and does not require special isolation chambers or mechanical disruption of the pancreatic tissue.
Microencaosulation Techniques
Microencapsulation of islet cells generally involves three steps: (a) generating microcapsules enclosing the islet cells (e.g., by forming droplets of cell-containing liquid alginate followed by exposure to a solution of calcium chloride to form a solid gel), (b) coating the resulting gelled spheres with additional outer coatings (e.g., outer coatings comprising polylysine and/or polyornithine) to form a semipermeable membrane; and (c) liquefying the original core gel (e.g., by chelation using a solution of sodium citrate). The three steps are typically separated by washings in normal saline.
A preferred method of microencapsulating pancreatic cells is the alginate-polyamino acid technique. Briefly, islet cells are suspended in sodium alginate in saline, and droplets containing islets are produced. Droplets of cell-containing alginate flow into calcium chloride in saline. The negatively charged alginate droplets bind calcium and form a calcium alginate gel. The microcapsules are washed in saline and incubated with poly-L-lysine or poly-L-ornithine (or combinations thereof); the positively charged poly-l-lysine and/or poly-L-ornithine displaces calcium ions and binds (ionic) negatively charged alginate, producing an outer poly-electrolyte membrane. A final coating of sodium alginate may be added by washing the microcapsules with a solution of sodium alginate, which ionically bonds to the poly-L-lysine and/or poly-L-ornithine layer. See U.S. Pat. No. 4,391,909 to Lim et al (all US patents referenced herein are intended to be incorporated herein in their entirety). This technique produces what has been termed a "single-wall" microcapsule. Preferred microcapsules are essentially round, small, and uniform in size. Wolters et al., J. Appli Biomater. 3:281 (1992).
When desired, the alginate-polylysine microcapsules can then be incubated in sodium citrate to solubilize any calcium alginate that has not reacted with poly-1-lysine, i.e., to solubilize the internal core of sodium alginate containing the islet cells, thus producing a microcapsule with a liquefied cell-containing core portion. See Lim and Sun, Science 210:908 (1980). Such microcapsules are referred to herein as having "chelated", "hollow" or "liquid" cores.
A "double-wall" microcapsule is produced by following the same procedure as for single-wall microcapsules, but prior to any incubation with sodium citrate, the microcapsules are again incubated with poly-1-lysine and sodium alginate.
Alginates are linear polymers of mannuronic and guluronic acid residues. Monovalent cation alginate salts, e.g., Na-alginate, are generally soluble. Divalent cations such as Ca++, Ba++ or Sr++ tend to interact with guluronate, providing crosslinking and forming stable alginate gels. Alginate encapsulation techniques typically take advantage of the gelling of alginate in the presence of divalent cation solutions. Alginate encapsulation of cells generally involves suspending the cells to be encapsulated in a solution of a monovalent cation alginate salt, generating droplets of this solution, and contacting the droplets with a solution of divalent cations. The divalent cations interact with the alginate at the phase transition between the droplet and the divalent cation solution, resulting in the formation of a stable alginate gel matrix being formed. A variation of this technique is reported in U.S. Pat. No. 5,738,876, wherein the cell is suffused with a solution of multivalent ions (e.g., divalent cations) and then suspended in a solution of gelling polymer (e.g., alginate), to provide a coating of the polymer.
Chelation of the alginate (degelling) core solubilizes the internal structural support of the capsule, may adversely affect the durability of the microcapsule, and is a harsh treatment of the encapsulated living cells. Degelling of the core may also cause leaching out of the unbound poly-lysine or solubilized alginate, resulting in a fibrotic reaction to the implanted microcapsule. The effect of core liquidation on glucose-stimulated insulin secretion by the encapsulated cells has been studied. Fritschy et al., Diabetes 40:37 (1991). The present inventors examined the function of islets enclosed in microcapsules that had not been subjected to liquefaction of the core (i.e., 'solid' or non-chelated microcapsules). It was found that culture of solid microcapsules prior to use enhanced the insulin response of the enclosed islets to glucose stimulation.
Alginate/polycation encapsulation procedures are simple and rapid, and represent a promising method for islet encapsulation for clinical treatment of diabetes. Many variations of this basic encapsulation method have been described in patents and the scientific literature. Chang et al., U.S. Pat. No. 5,084,350 discloses microcapsules enclosed in a larger matrix, where the microcapsules are liquefied once the microcapsules are within the larger matrix. Tsang et al., U.S. Pat. No. 4,663,286 discloses encapsulation using an alginate polymer, where the gel layer is cross-linked with a polycationic polymer such as polylysine, and a second layer formed using a second polycationic polymer (such as polyornithine); the second layer can then be coated by alginate. U.S. Pat. No. 5,762,959 to Soon-Shiong et al. discloses a microcapsule having a solid (non-chelated) alginate gel core of a defined ratio of calcium/barium alginates, with polymer material in the core.
U.S. Pat. Nos. 5,801,033 and 5,573,934 to Hubbell et al. describe alginate/polylysine microspheres having a final polymeric coating (e.g., polyethylene glycol (PEG)); Sawhney et al., Biomaterials 13:863 (1991) describe alginatelpolylysine microcapsules incorporating a graft copolymer of poly-1-lysine and polyethylene oxide on the microcapsule surface, to improve biocompatibility; U.S. Pat. No. 5,380,536 describes microcapsules with an outermost layer of water soluble non-ionic polymers such as polyethylene(oxide). U.S. Pat. No. 5,227,298 to Weber et al. describes a method for providing a second alginate gel coating to cells already coated with polylysine alginate; both alginate coatings are stabilized with polylysine. U.S. Pat. No. 5,578,314 to Weber et al. provides a method for microencapsulation using multiple coatings of purified alginate.
U.S. Pat. No. 5,693,514 to Dorian et al. reports the use of a non-fibrogenic alginate, where the outer surface of the alginate coating is reacted with alkaline earth metal cations comprising calcium ions and/or magnesium ions, to form an alkaline earth metal alginate coating. The outer surface of the alginate coating is not reacted with polylysine.
U.S. Pat. No. 5,846,530 to Soon-Shiong describes microcapsules containing cells that have been individually coated with polymerizable alginate, or polymerizable polycations such as polylysine, prior to encapsulation.
The methods of the present invention are intended for use with any microcapsule that contains living cells secreting a desirable biological substance (preferably pancreatic cells and more preferably islet cells), where the microcapsule comprises an inner gel or liquid core containing the cells of interest, or a liquid core containing the cells of interest, bounded by a semi-permeable membrane surrounding the cell-containing core. The inner core is preferably composed of a water-soluble gelling agent; preferably the water-soluble gelling agent comprises plural groups that can be ionized to form anionic or cationic groups. The presence of such groups in the gel allows the surface of the gel bead to be cross-linked to produce a membrane, when exposed to polymers containing multiple functionalities having a charge opposite to that of the gel.
Cells suspended in a gellable medium (such as alginate) may be formed into droplets using any suitable method as is known in the art, including but not limited to emulsification (see e.g., U.S. Pat. No. 4,352,883), extrusion from a needle (see, e.g., U.S. Pat. No. 4,407,957; Nigam et al., Biotechnology Techniques 2:271-276 (1988)), use of a spray nozzle (Plunkett et al., Laboratory Investigation 62:510-517 (1990)), or use of a needle and pulsed electrical electrostatic voltage (see, e.g., U.S. Pat. Nos. 4,789,550; 5,656,468).
The water-soluble gelling agent is preferably a polysaccharide gum, and more preferably a polyanionic polymer. An exemplary water-soluble gelling agent is an alkali metal alginate such as sodium alginate. The gelling agent preferably has free acid functional groups and the semi-permeable membrane is formed by contacting the gel with a polymer having free amino functional groups with cationic charge, to form permanent crosslinks between the free amino acids of the polymer and the acid functional groups. Preferred polymers include polylysine, polyethylenimine, and polyarginine. A particularly preferred microcapsule contains cells immobilized in a core of alginate with a poly-lysine coating; such microcapsules may comprise an additional external alginate layer to form a multi-layer alginate-polylysine-alginate microcapsule. See U.S. Pat. No. 4,391,909 to Lim et al, the contents of which are incorporated by reference herein in their entirety.
When desired, the microcapsules may be treated or incubated with a physiologically acceptable salt such as sodium sulfate or like agents, in order to increase the durability of the microcapsule, while retaining or not unduly damaging the physiological responsiveness of the cells contained in the microcapsules. By "physiologically acceptable salt" is meant a salt that is not unduly deleterious to the physiological responsiveness of the cells encapsulated in the microcapsules. In general, such salts are salts that have an anion that binds calcium ions sufficiently to stabilize the capsule, without substantially damaging the function and/or viability of the cells contained therein. Sulfate salts, such as sodium sulfate and potassium sulfate, are preferred, and sodium sulfate is most preferred. The incubation step is carried out in an aqueous solution containing the physiological salt in an amount effective to stabilize the capsules, without substantially damaging the function and/or viability of the cells contained therein as described above. In general, the salt is included in an amount of from about 0.1 or 1 milliMolar up to about 20 or 100 millimolar, most preferably about 2 to 10 millimolar. The duration of the incubation step is not critical, and may be from about 1 or 10 minutes to about 1 or 2 hours, or more (e.g., over night). The temperature at which the incubation step is carried out is likewise not critical, and is typically from about 4 degrees Celsius up to about 37 degrees Celsius, with room temperature (about 21 degrees Celsius) preferred.
When desired, liquefaction of the core gel may be carried out by any suitable method as is known in the art, such as ion exchange or chelation of calcium ion by sodium citrate or EDTA. Microcapsules useful in the present invention thus have at least one semipermeable surface membrane surrounding a cell-containing core. The surface membrane permits the diffusion of nutrients, biologically active molecules and other selected products through the surface membrane and into the microcapsule core. The surface membrane contains pores of a size that determines the molecular weight cut-off of the membrane. Where the microcapsule contains insulin-secreting cells, the membrane pore size is chosen to allow the passage of insulin from the core to the external environment, but to exclude the entry of host immune response factors.
As used herein, a "poly-amino acid-alginate microsphere" refers to a capsule of less than 2 mm in diameter having an inner core of cell-containing alginate bounded by a semi-permeable membrane formed by alginate and poly-1-lysine. Viable cells encapsulated using an anionic polymer such as alginate to provide a gel layer, where the gel layer is subsequently cross-linked with a polycationic polymer (e.g., an amino acid polymer such as polylysine. See e.g., U.S. Pat. Nos. 4,806,355, 4,689,293 and 4,673,566 to Goosen et al.; U.S. Pat. Nos. 4,409,331, 4,407,957, 4,391,909 and 4,352,883 to Lim et al.; U.S. Pat. Nos. 4,749,620 and 4,744,933 to Rha et al.; and U.S. Pat. No. 5,427,935 to Wang et al. Amino acid polymers that may be used to encapsulate islet cells in alginate include the cationic amino acid polymers of lysine, arginine, and mixtures thereof.
Culture of Isolated Cells
Generally, pancreatic islets are isolated by collagenase digestion of pancreatic tissue. This process involves subjecting the islet cells to a period of hypoxia which is then followed by reoxygenation. Hypoxia-reoxygenation produces an injury that is linked to excessive production of oxygen free radicals which impair the function, and cause the death, of islet cells, particularly those isolated from the pancreas of large mammals such as pigs and humans.
The present inventors have determined that culture of isolated islets or islet cells prior to microencapsulation is beneficial. The islets are cultured according to known cell culture techniques for a period of at least 3 hours, more preferably from 12-36 hours, and more preferably from 18-24 hours, in a culture medium containing an antioxidant compound. More preferably, the culture medium contains any one of, or any combination of, the following: an antioxidant, an anti-cytokine, an anti-endotoxin, and an antibiotic.
Culture of isolated pancreas cells to improve glucose-stimulated insulin secretion may utilize any suitable anti-oxidant as is known in the art. As used herein, an antioxidant is a compound that neutralizes free radicals or prevents or suppresses the formation of free radicals. Particularly preferred are molecules including thiol groups such as reduced glutathione (GSH) or its precursors, glutathione or glutathione analogs, glutathione monoester, and N-acetylcysteine. Other suitable anti-oxidants include superoxide dismutase, catalase, vitamin E, Trolox, lipoic acid, lazaroids, butylated hydroxyanisole (BHA), vitamin K, and the like. Glutathione, for example, may be used in a concentration range of from about 2 to about 10 mM. See, e.g., U.S. Pat. Nos. 5,710,172; 5,696,109; 5,670,545.
Culture of isolated pancreas cells to improve glucose-stimulated insulin secretion may utilize any suitable antibiotic as is known in the art. Suitable antibiotics include penicillins, tetracyclines, cephalosporins, macrolides, β-lactams and aminoglycosides; examples of such suitable antibiotics include streptomycin and amphotericin B.
Culture of isolated pancreas cells to improve glucose-stimulated insulin secretion may utilize any suitable anti-cytokine as is known in the art. Cytokines are proteins secreted by many different cell types, that regulate the intensity and duration of immune responses. Cytokines include various growth factors, colony-stimulating factors, interleukins, lymphokines, monokines and interferons. Anti-cytokines are compounds that prevent or suppress the function of cytokines. Suitable anti-cytokines for use in culturing islet cells include dimethylthiourea (10 mM), citiolone (5 mM), pravastatin sodium (PRAVACHOL®, 20 mg/kg), L-NG-monomethylarginine (L-NMMA, 2 mM), lactoferrin (100 μg/ml), 4-methylprednisolone (20 μg/ml), and the like.
Culture of isolated pancreas cells to improve glucose-stimulated insulin secretion may utilize any suitable anti-endotoxin as is known in the art. Endotoxins are bacterial toxins, complex phospholipid-polysaccharide molecules that form a part of the cell wall of a variety of Gram-negative bacteria. Anti-endotoxins are compounds that destroy or inhibit the activity of endotoxins. Endotoxins are intracellular toxins, and are complex phospholipid-polysaccharide macromolecules that form a part of the cell wall of a variety of Gram-negative bacteria, including enterobacteria, vibrios, brucellae and neisseriae. Suitable anti-endotoxins for use in culturing islet cells include L-NG-Monomethylarginine (L-NMMA, 2 mM), lactoferrin (100 μg/ml), N-acetylcysteine (NAC, 1 mM), adenosine receptor antagonists such as bamiphylline (theophylline) and anti-lipopolysaccharide compounds such as echinomycine (10 nM), and the like.
Cryopreservation of Cells
Mammalian tissue remains viable in vitro only for short periods of time, usually days. Loss of islet cells suitable for transplantation may be avoided by viable cryopreservation and cold storage of the cells. The present inventors determined that microencapsulated islet cells respond poorly to cryopreservation. However, cryopreservation of naked (unencapsulated) islet cells did not adversely affect their later function in microcapsules when the cells were first cryopreserved, then thawed and microencapsulated. Frozen and thawed microencapsulated islets responded poorly to glucose stimulation; in comparison, 'naked' islet cells that were cryopreserved and then thawed retained their ability to respond to glucose stimulation and were suitable for microencapsulation. Islet cells can thus be preserved by cryopreservation, thawed and microencapsulated just prior to use.
Methods of cryopreservation are well known in the art. In general terms, cryopreservation of animal cells involves freezing the cells in a mixture of a growth medium and another liquid that prevents water from forming ice crystals, and then storing the cells at liquid nitrogen temperatures (e.g., from about −80 to about −196° C.).
An aspect of the present invention is the cryopreservation of isolated mammalian cells in a cryopreservation medium containing an antioxidant, followed by microencapsulation of the cells prior to in vivo implantation. A preferred embodiment of the present invention is the cryopreservation of isolated islets or islet cells in a cryopreservation medium containing an antioxidant as described herein, followed by microencapsulation prior to in vivo implantation.
More preferably, the cells are cryopreserved in a medium containing at least one of, or a combination of, the following: an antioxidant, an anti-cytokine, an anti-endotoxin, and an antibiotic (each as described above). Preferably, the cells are cryopreserved in a medium containing at least one each of an antioxidant, an anti-cytokine, an anti-endotoxin, and an antibiotic (each as described above).
Culture of Microspheres.
The present inventors studied the response to glucose of islet-containing microcapsules, and found that culture of such microcapsules prior to use enhanced subsequent glucose-stimulated insulin production. The enclosed islets responded better to a glucose challenge than islets contained in fresh (non-cultured) microcapsules.
Culture of microencapsulated cells is carried out in a manner similar to the culture of isolated cells, as described herein and as generally known in the art. Accordingly, a method of the present invention is the culture of microcapsules (with either solid or liquid cores containing living cells) prior to implantation in vivo, to enhance or improve the ability of the microcapsule to produce a desired cell secretory product. A particularly preferred embodiment is the culture, prior to implantation, of gelled or solid-core alginate-polylysine microcapsules containing pancreatic islets or islet cells. Microcapsules are cultured for a period of at least 3 hours, more preferably from 12 to 36 hours, or 18 to 24 hours, prior to implantation.
Preferably the microcapsules are cultured in a medium containing at least one of, or a combination of, the following: an antibiotic, an antioxidant, an anticytokine, and an antiendotoxin (as described above). More preferably, the microcapsules are cultured in a medium containing at least one each of an antioxidant, an anti-cytokine, an anti-endotoxin, and an antibiotic (each as described above).
As noted above, microencapsulated islet cells are also an aspect of this invention. In general, such microencapsulated islet cells comprise a microcapsule containing living pancreatic islet cells therein, the microcencapsulated islet cells exhibiting a weight gain of not more than 1, 5 or 10 percent by weight over a period of one month in physiological saline solution at 37 degrees Celsius (exhibiting the durability thereof) and exhibiting at least 150, 200 or 250 percent basal insulin secretion in response to 16.7 millimolar glucose challenge in Krebs-Ringer physiological solution at pH 7.4 after said period of one month. Such microencapsulated cells may be produced by the procedures described herein, preferably including the step of incubating them with a physiologically acceptable salt to enhance the durability or stability of the capsule as described above.
Encapsulated islet cells produced according to the present invention may be transplanted into subjects as a treatment for insulin-dependent diabetes; such transplantation may be into the peritoneal cavity of the subject. An amount of encapsulated islet cells to produce sufficient insulin to control glycemia in the subject is provided by any suitable means, including but not limited to surgical implantation and intraperitoneal injection. The International Islet Transplant Registry has recommended transplants of at least 6,000 islets, equivalent to 150 μm in size, per kilogram of recipient body weight, to achieve euglycemia. However, it will be apparent to those skilled in the art that the quantity of microcapsules transplanted depends on the ability of the microcapsules to provide insulin in vivo, in response to glucose stimulation. One skilled in the art will be able to determine suitable transplantation quantities of microcapsules, using techniques as are known in the art.
As used herein, materials that are intended to come into contact with biological fluids or tissues (such as by implantation or transplantation into a subject) are termed "biomaterials". It is desirable that biomaterials induce minimal reactions between the material and the physiological environment. Biomaterials are considered "biocompatible" if, after being placed in the physiological environment, there is minimal inflammatory reaction, no evidence of anaphylactic reaction, and minimal cellular growth on the biomaterial surface. Upon implantation in a host mammal, a biocompatible microcapsule does not elicit a host response sufficient to detrimentally affect the function of the microcapsule; such host responses include formation of fibrotic structures on or around the microcapsule, immunological rejection of the microcapsule, or release of toxic or pyrogenic compounds from the microcapsule into the surrounding host tissue.
The term "encapsulate" as used herein refers to the containment of a cell or cells within a capsule delineated by a physical barrier (i.e., a barrier that reduces or controls the permeability of the capsule). The term "microcapsule" or "microsphere" as used herein refers to a structure containing a core of biological substance (such as cells) in an aqueous medium, surrounded by a semi-permeable membrane, and having a diameter of no more than 2 mm. Preferably, microspheres are from about 3μm to about 2 mm in diameter. More preferably, microcapsules range from about 50μm to about 1,000 μm in diameter, or from about 300 μm to about 500 μm in diameter. Depending on the method of microencapsulation used, it will be apparent that the microcapsule wall or membrane may also contain some cells therein.
As used herein, culture refers to the maintenance or growth of cells on or in a suitable nutrient medium, after removal of the cells from the body. Suitable nutrient culture media are readily available commercially, and will be apparent to those skilled in art given the cell type to be cultured.
The term "cells" as used herein refers to cells in various forms, including but not limited to cells retained in tissue, cell clusters (such as pancreatic islets or portions thereof), and individually isolated cells.
Male Sprague-Dawley rats weighing 200-300 g were obtained (Charles River). Collagenase (Type P) was obtained from Boehringer Mannheirn Co. (Indianapolis, Ind.). Highly purified bovine serum albumin (BSA, fraction V) free of fatty acids and insulin-like activity and all chemicals for buffer and encapsulation Solution were obtained from Sigma Chemical Co. (St. Louis, Mo.). Monoiodinated 125I-insulin was obtained from New England Nuclear (Boston, Mass.).
Islet Isolation
Islets were isolated by collagenase digestion: (Lacy et al., Diabetes 16:35 (1967)). Rats were anesthetized with intraperitoneal pentobarbital, laparotomy was performed, and the pancreatic duct was cannulated. The pancreas was distended by infusing 15 ml Hank's Balanced Salt Solution (HBSS). Pancreatectomy was performed, and the pancreas was digested in collagenase solution (1.5 mg/ml) at 37° C. for 25 minutes. The digested tissue was filtered through a mesh and then washed several times in chilled HBSS and placed under a dissecting microscope. At least 400 islets/experiment were purified by being handpicked and placed in chilled HBSS until microencapsulation.
Islets were suspended in 1.4% sodium alginate and placed in a droplet generator adapted from that of Walters et al., J. Appl. Biomater. 3:281 (1992). Droplets generated from islets suspended in the alginate solution were collected in a funnel containing 1.1% CaCl2, where they gelled. The resulting microcapsules were washed with normal saline (NS), incubated with 0.05% poly-L-lysine, washed again with NS, incubated with alginate, and washed a final time with NS. These islet-containing microcapsules were then split into three groups. Group II islets were set aside for immediate perifusion. Group III islets were also set aside without chelation, for 24 hours of culture in RPMI 1640 at 37° C. prior to perifusion. In Group I islets, the alginate core of the microcapsule was liquefied by incubation with a chilled 55 mM sodium citrate solution for seven minutes, to chelate the central alginate-Ca2+ core.
Unencapsulated and microencapsulated islets were perifused by a method previously described (Garfinkel et al., J. Surg. Res. 52:328 (1992)). In each case, 30 islets were handpicked under a stereomicroscope and placed (10/chamber) in a plastic flow-through perifusion chamber (Millipore Corp.). Islets were perifused for 1 hour at 37° C., at the rate of 1 ml/min, with a modified Krebs-Ringer-bicarbonate (KRB) solution (Opara et al., Endocrinology 130:657 (1992)), containing 1% BSA and a basal 3.3 mM (60 mg/dl) glucose, maintained at pH 7.4 by continuous gassing with 95% O2/5%CO2, in an unsealed reservoir. All perifusion experiments were conducted with triplicate chambers housing islets from the same preparation, with each chamber receiving microencapsulated islets prepared under one of the three conditions described above.
After the 1-hour perifusion, basal effluent samples were periodically collected on ice for 20 minutes before islets were then perifused with the KRB solution containing 16.7 mM glucose (300 mg/dl) for 30 minutes, during which samples were again collected at intervals. Following the 16.7 mM glucose stimulation, a washout perifusion with the 3.3 mM glucose buffer was performed for 20 minutes with sample collection. Solutions were changed using a stopcock and all effluent perifusate samples obtained throughout the study were stored frozen at −20° C. until radioimmunoassay for insulin (Herbert et al., J. Clin. Endocrinol. Metab. 25:1375 (1965)).
Data Analysis
Results are expressed as mean±SEM. Results were plotted as rates of insulin secretion (pg insulin/10 islets/minute) versus time. Peak rates of insulin secretion during the first stimulatory phase were compared with basal rates within the same group, using a Student's t test. For comparisons among groups, an analysis of variance (ANOVA) computer program was used and depending on the outcome of ANOVA, the Bonferroni correction was made. In all cases, significance was defined as P<0.05.
Control Islets
Unencapsulated islets were first perifused with the high 16.7 mM glucose to determine the normal isolated islet response to stimulation with this concentration of glucose. The results are presented in FIG. 1 (representing four experiments) with a mean basal rate of insulin secretion of 2650±40 pg/10 islets/minute.
When the glucose concentration in the perifusate was raised to 16.7 mM, insulin response of the islets was biphasic, with a mean rate of 5120±310 pg/10 islets/minute (P<0.002 versus basal), which subsequently returned to basal upon switching back to basal perifusion (FIG. 1).
Experimental Islets Groups I, II, and III
Group I islets (liquefied core
microcapsules) responded similarly as compared to control islets with a biphasic response to the stimulatory concentration of glucose (FIG. 2). In six experiments, insulin secretion increased from a basal rate of 3005±647 pg/10 islets/minute to a stimulated rate of 5164±1428 pg/10 islets/minute (P<0.05), during the first phase secretion in response to stimulation with 16.7 mM glucose. As noted in control unencapsulated islets (FIG. 1), in Group I islets, the rate of insulin secretion returned to basal after withdrawal of the stimulatory 16.7 mM glucose (FIG. 2).
In contrast, Group II islets (gelled core microcapsules, n=6) had a lower basal rate of insulin secretion (1740±219 pg/10 islets/minutes, P<0.05 vs. Group I basal). These islets did not significantly respond to 16.7 mM glucose stimulation (FIG. 3).
Group III islets (FIG. 4), which represent cultured gelled core microcapsules, also had a lower basal rate of insulin secretion (1375±161 pg/10 islets/minutes) than control islets, as well as those in Groups I and II (P<0.05). However, in contrast to the freshly perifused gelled core microcapsules (Group II), the islets in the cultured gelled-core microcapsules (Group III) had a small but significant increase in their rate of insulin secretion in response to 16.7 mM glucose stimulation (P<0.05, n=6) as shown in FIG. 4, and in Table 1 which summarizes the data from the control and the three experimental groups.
The function of cryopreserved, microencapsulated islet cells, and the stability of the capsule, was studied.
Islets were isolated from Sprague-Dawley rats by collagenase digestion of pancreatic tissue, and microencapsulated in polylysine-alginate using a syringe-air-droplet generator. Aliquots of fresh unencapsulated (group 1) and encapsulated (group 2) islets were taken for determination of glucose stimulation of insulin secretion (GSIS). The remaining portions of unencapsulated (group 3) and encapsulated (group 4) were then frozen for 1-3 days at −80° C. using standard techniques. After thawing, GSIS was also tested in these two groups of islets, using a perifusion model.
GSIS in groups 1, 2 and 3 were comparable, averaging a 2-fold stimulation (p<0.05) within 10 minutes of raising the glucose concentration from 3.3 mM (60 mg/dL) to 16.7 mM (300 mg/dL). In group 4, a mean ±SEM of 14.5±0.3% of the capsules were broken after thawing and glucose-stimulated insulin secretion was not observed.
These studies indicate that isolated islets should be cryopreserved unencapsulated and then microencapsulated just prior to use in transplantation.
The present inventors further studied the effects of culture on microencapsulated islet function. The response to glucose of freshly prepared encapsulated islets (non-cultured microcapsules) was compared to that of cultured encapsulated islets, using a validated in vitro perifision system.
Isolated rat islets were purified by handpicking under a stereomicroscope, and then microencapsulated in alginate-polylysine-alginate using a syringe-air-droplet generator, prior to chelation of the inner alginate core. The encapsulated islets were either tested immediately (control) or cultured for 24 hours prior to perifusion with different concentrations of glucose.
In control (non-cultured) islet microcapsules, the mean±SEM rate of insulin secretion increased from a basal level of 815±5 to 1668±75 pg/6 islets/minute (p<0.001, n=5) within ten minutes of raising the glucose concentration from 60 mg/dL (basal) to 16.7 mM (300 mg/dL).
In cultured islet microcapsules, the mean±SEM rate of insulin secretion increased from a basal of 928±46 to 2433±200 pg/6 islets/minute (p<0.001, n=7).
The degree of stimulation in the cultured microcapsules (262±20% of basal, p<0.05) was greater than that in control (non-cultured) microcapsules (205±9% of basal). These studies indicate that primary culture of encapsulated islet cells enhances their function.
In an anesthetized pig, the pancreas is perifused with University of Wisconsin Solution (DuPont) and pancreatectomy without uncination is performed. The pancreatic ducts are distended with HBSS containing 1 mM Trolox (an antioxidant), 1.5 mg/ml collagenase and 10,000 units DNAse 1. The distended pancreas is placed on ice and digested for 30 minutes, then the pancreas with digestion medium is incubated at 37° C. for twenty minutes prior to shaking by hand for one minute. Digested tissue is then filtered through mesh, then washed in RPMI 1640 with low speed centrifugation. The islet cells are separated on a Ficoll gradient with a COBE automatic cell separator. The isolated naked islet cells are then incubated overnight (18-24 hours) in RPMI 1640 culture media containing at least one each of an antibiotic, antioxidant, anticytokine, and antiendotoxin. A representative medium contains 5 cc of a mixture of streptomycin/penicillin (per 100 ml culture medium), 10 mM dimethylthiourea, 5 mM citiolone, 2 mM L-NMMA, and 10 mM GSH
The naked (unencapsulated) islet cells are then cryopreserved (frozen at a minimum temperature of −80° C.) in an antioxidant/cryopreservation cocktail, consisting of medium 199, 2M dimethylsulfoxide, 10% pig serum, 0.7 mM L-glutamine, and 25 mM HEPES.
The frozen islets are then subjected to rapid thawing at 37° C., placed in Kelco's alginate (1000 islets/ml). Kelco's alginate contains 1.4% sodium alginate, 25 mM HEPES, 118 mM NaCl, 2.5 mM MgCl2, at pH=7.4. The alginate is then placed in a syringe and, using a droplet generator, droplets are produced that fall into a funnel filled with calcium solution (1.1% CaCl2, 10 mM HEPES, pH=7.4) to gel the droplets.
The gelled droplets are then washed in saline, incubated in 0.05% Poly-L-Lysine for six minutes, saline washed, incubated in sodium alginate (0.06%) for four minutes, saline washed twice, incubated in 50 mM sodium citrate for seven minutes, and saline washed twice to produce chelated microencapsulated islet cells of 400-700 μM in diameter.
The chelated microencapsulated islet cells are cultured for 3-24 hours in a cocktail containing five ingredients: 10 Mm dimethylurea, 5 mM citiolone, 2 mM N-monomethyl-L-arginine; 10 mM GSH; 5 mM L-glutamine.
Animal subjects (baboons) are rendered diabetic by partial (70%) pancreatectomy and intravenous administration of streptozotocin (65 mg/kg), and are treated with 20 mg/ml 4-methylprednisone (antiendotoxin). Microcapsules are injected intraperitoneally; from about 5,000 to about 15,000 encapsulated islets/kg body weight.
After transplantation, the animals' blood glucose levels are monitored daily.
Alginate is an anionic polysaccharide composed of variable homopolymeric regions of mannuronic acid and guluronic acid interspaced with regions of alternating blocks (Smidsrod et al., Trends in Biotechnol. 8: 71-78 (1990)). Cross-linking with divalent cations such as Ba++ and Ca++ occurs between the carboxyl groups of the guluronic and mannuronic acid chains. However, some of the carboxyl groups remain free even after the cross-linking procedure. This situation allows for unbound positively charged divalent cations to attach to the remaining free negatively charged carboxyl groups within the alginate bead matrix. Also, negatively charged molecules from the HEPES buffer used in the cross-linking step may be entrapped in the alginate beads during the process of cross-linking and the molecules may also bind divalent cations. Therefore, such events produce an undesirable excess of divalent cations being associated with the alginate matrix. The cations are likely undesirable in that they attract water molecules and therefore cause the formation of colloid osmotic pressure. The effect will likely result in weakness, swelling, and possible rupture in such microcapsules.
To prevent the possibility of this phenomenon of alginate microcapsule weakness, swelling, and rupture, the alginate beads are washed with sodium sulphate, to induce a chemical reaction designed to eliminate the divalent cation problem, as described above. It was discovered that the incubation of chelated and saline-washed islet microcapsules, as described in previous examples above, in 6 mM sodium sulfate for 30 minutes at room temperature, followed by washing in normal saline (so that excess divalent cations are bound by the sulphate anion), and then culturing at 37 degrees Celsius, results in a significant enhancement of durability (see FIG. 5). Islets contained in the sulfate treated, and more durable microcapsules, are as functionally viable, with regard to their physiological profiles, as those in microcapsules not treated with the sulfate wash step (see FIG. 6).
The foregoing is illustrative of the present invention, and is not to be construed as limiting thereof. The invention is defined by the following claims, with equivalents of the claims to be included therein.
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Illustration: Street gangs, cool guys, greedy pigs and jazz – step into the world of Gaurab Thakali
Stumbling across Gaurab's work is like when someone brings their cool, good looking cousin to a party full of people you see all the time; the whole room goes "Well, hello-o!" Gaurab is still studying (yes, still studying) illustration at Camberwell University. His naive, unique-paletted work is inspired by the lost heroes of jazz, street fights, and the kind of safe but scrappy, inner-city atmosphere prevalent in Hey, Arnold!. Gaurab's portfolio is one of the most unique I've seen in ages, so considering he hasn't even graduated yet, we're keeping a very firm eye on him indeed.


text:
Ethan Klingsberg
Meredek Street high in the hills overlooking Budapest,
construction of attractive homes for the first wave of post-Communist
nouveau riche is busily under way. One old socialist-style
block home stands out on the street: the heavily fortified
No. 38. Currently, the speaker of Hungary's Parliament lives
there. But in the late 1950s this was home to Noel Field,
one of the Soviet bloc's prized asylum-seekers from the United
States. Nobody on Meredek Street claims to remember Field.
But detailed memories of Noel Field's life exist in Budapest
in decades-old secret-police files at the Historical
Archive of the Hungarian Interior Ministry. Field was a Communist
sympathizer and perhaps a minor Soviet agent with a troubled
career on both sides of the Iron Curtain branded a
Soviet spy by Whittaker Chambers before the House Un-American
Activities Committee in the United States, imprisoned as an
American spy in Hungary. An op-ed in the October 15 New
York Times and an article in Commentary this past
April claimed that those Hungarian files on Field contain
something explosive: "unimpeachable" evidence that
will "seal the case against Alger Hiss."
both the Times and Commentary pieces, author
Sam Tanenhaus relies upon the unpublished findings of a 34-year-old
Hungarian historian, Maria Schmidt, who in the course of research
in the secret-police archives uncovered statements by Field
that seemed directly to implicate Hiss in spying.
presented her findings which she had already discussed
on Hungarian radio in New York on October 11, at a
seminar sponsored by New York University's Institute for the
Humanities. After decades of trials, books and investigative
reporting, is there finally definitive proof that Alger Hiss
worked for the Soviet Union in the 1930s, when he was a U.S.
government employee?
after Commentary's "Hiss: Guilty as Charged"
article appeared, The Nation contacted me in Budapest,
where I had recently directed an international conference
on freedom of information and the uses of Communist-era archives.
Was there any way to review Maria Schmidt's sources? Through
the Budapest conference I had met Gabor Baczoni, director
of the Interior Ministry archive. After a twelve-week application
process graciously facilitated by Baczoni, I became the only
foreign researcher to review Noel Field's 2,500-page dossier,
and in particular those documents relating to Hiss. In the
words of Baczoni, I "saw everything in the Interior Ministry's
Noel Field dossier that Maria Schmidt saw." What I saw
turns out to provide a case study in why this new game of
using uncritical readings of Communist secret-police files
to make definitive historical pronouncements is misguided
and does not serve the interests of truth.
Strange Life of Noel Field
Harvard graduate, Field enthusiastically dedicated his life
until 1947 to public service in the West, yet he ended up
living out his days in silence and depression in an oppressive
Soviet satellite. A friend of Communists during World War
II relief efforts, he would eventually fall victim to Stalinist
interrogators, who extracted false and distorted statements
from him about his Communist Party contacts for use as the
basis for hundreds of purges and executions throughout the
Soviet bloc.
was a social acquaintance of Alger Hiss in Washington, D.C.,
during 1935-36. Field worked for the State Department at the
time, while Hiss worked first for the Agriculture Department
and then moved over to the Solicitor General's office. Field
left State in 1936 and headed off for Europe to work for the
League of Nations; from 1941 to 1947 he served in Europe with
the Unitarian Service Committee. After 1947 he traveled around
Eastern Europe for two years as an unsuccessful freelance
writer or "student of popular democracy," depending
on whom he was talking to. On August 27, 1948, Whittaker Chambers
linked Noel Field to his own accusations against Alger Hiss,
claiming that Hiss had tried to "draw Field in"
to Hiss's alleged Communist espionage cell, only to discover
that Field "was already a Communist working in another
apparatus." As it turned out, HUAC would not be the only
source of trouble for Field. In 1949 he was arrested by Communist
police in Prague on charges that he was an "American
master-spy" attempting to undermine the Soviet Union.
He spent the next five years in Hungarian prisons, where he
was tortured and eventually went on a hunger strike. On his
release he requested asylum in Hungary and settled in Budapest.
He died in 1970; his ashes now rest in a special crematorium
depository for Communist heroes in Budapest.
Hungarian Interior Ministry's Noel Field dossier spans the
period 1945-65 and includes Field's personal papers prior
to and following his imprisonment; records of his interrogations
by the secret police; and records of his voluminous communications
with both Hungarian and Soviet authorities during his imprisonment.
If I had cut out all the references to Alger Hiss from the
thousands of pages in the Field dossier and then pasted them
together, they would constitute about four pages. It is these
papers that Maria Schmidt and her American chronicler, Sam
Tanenhaus, claim "seal the case" against Hiss. According
to Schmidt, the damning evidence consists of: (1) correspondence
between Field and Hiss; (2) expressions of anxiety by Field
concerning Whittaker Chambers's testimony; (3) Field's "musings
on the coincidence" that he and Hiss were released from
prison at the same time; and (4) direct statements by Field
to his Hungarian interrogators that Hiss was a fellow Communist
as well as a Soviet agent who competed with another Communist
cell to recruit Field for espionage.
us consider each category of evidence in turn. The first three
can be dealt with quickly.
Anxiety and Musings
first exchange of letters between Field and Hiss to be found
in the Hungarian archives dates from the fall of 1948, the
time of Whittaker Chambers' HUAC testimony. Hiss wrote on
October 14, 1948, to inform Field, living in Europe and without
access to American newspapers, that Field was "irresponsibly
smeared" in Chambers' testimony. On its face Hiss's letter
appears to be simply from one former colleague to another.
(I recently asked Hiss about this old letter. "It was
a warning to a friend," he says. "I was probably
also counting on Noel to help counter Chambers' testimony.")
November 2, 1948, Field replied:
as Chambers's recorded fabrications concerning me are, in
the main, based on his alleged conversations with you, I take
it there is no point in my dignifying them with any public
denial and that your libel action will automatically dispose
of them....I need hardly tell you how angered and outraged
I was over the irresponsible allegations made against you.
Your testimony fully harmonizes with the memory I had of you
during our all-too-brief acquaintance in Washington. While
my views, as I recall, were somewhat to the left of yours,
I always particularly admired you as an embodiment of the
best Oliver Wendell Holmes tradition and as a man of unusual
integrity in both his private and his official life.
I trust you will receive full satisfaction in your libel
I trust you will receive full satisfaction in your libel
after Field's release from prison in 1954, he wrote in response
to a letter from Hiss's attorney sent six years earlier, after
Field was first jailed. Hiss's attorney had requested Field's
"personal opinion" on "being mentioned"
in the testimony of witness Hede Massing during Hiss's perjury
prosecution. Massing had claimed Hiss tried to recruit Field
for a Communist cell. In his reply Field explained that he
had not answered earlier due to his imprisonment; then he
dismisses Hede Massing's testimony as "nothing but a
lie. . . I got to know Alger Hiss as a liberal without communist
attachments, but with a commitment to peace and the promotion
of understanding with whose wife we [Field and his wife] had
spent some time as Quakers in the past." Field goes on
to regret that his own "ill fame" in the United
States, arising from his decision to request asylum in a Communist
country, probably reduces the usefulness of his declaration.
of this seems to be much of a smoking gun. On its face, this
correspondence presumes Hiss's innocence. In the Times,
Tanenhaus propounded the theory that Field's post-release
letters must have been part of a Hungarian campaign to protect
Alger Hiss. While there can be no doubt that the Hungarian
secret police screened all of Field's letters, the secret-police
commentary on Fields' post-release correspondence in the dossier
fails to substantiate Tanenhaus's theory. In particular, the
secret police report on Field's 1955 letter to Hiss simply
According to our documents, Alger Hiss was arrested
by the American authorities. First he was accused of Communist
activity, then of espionage for the Soviet Union and he
was imprisoned. He was discharged from prison roughly when
the Fields were released too. He lives in the States now.
According to our documents, Alger Hiss was arrested
by the American authorities. First he was accused of Communist
activity, then of espionage for the Soviet Union and he
was imprisoned. He was discharged from prison roughly when
the Fields were released too. He lives in the States now.
the officials overseeing Field knew very little about Alger
air of innocence similarly characterizes Field's recorded
anxiety over Chambers's HUAC testimony. A week after his 1948
letter to Hiss, Field sent a note to former State Department
colleague Laurence Duggan lamenting the effect that Chambers'
HUAC testimony might have on his writing career. "My
first reaction was to explode with as audible a yell as I
could produce from these distant lands....I am only too aware
of the fact that my publishing aims whether in periodicals
or book form have hardly been advanced by the type
of publicity my name has gotten." Field's expressions
of fear never concede guilt and may have just been well-founded
concerns for his career, reputation and good friend.
Tanenhaus's two articles make much of a Hungarian secret police
record suggesting that Field noted a parallel between his
release from the Hungarian prison and Hiss's release in the
United States. When I read the actual document, however, I
discovered that this was yet another instance in which a document
that sounds incriminating turns out to be potentially insignificant.
On November 18, 1954, a Hungarian agent noted that Field commented
on the "wonderful coincidence" that he and his "old
friend Alger Hiss" had been released at about the same
time. Both the Hungarian agent and Field refrain from making
any judgment whatsoever about the veracity of Hiss's alleged
connection with Communism.
Prison Statements
Field dossier's most sensational references to Alger Hiss
appear in Field's prison statements. While Hede Massing was
testifying in 1949 at the Hiss trial that Field was a Soviet
agent, the Hungarian secret police had taken Field into custody
and were interrogating him about his alleged efforts to undermine
the Soviet Union on behalf of American intelligence. During
five years in prison he would produce over 1,000 pages of
prison statements in the form of petitions for release, "autobiographies"
and answers to interrogations.
reports by the Hungarian secret police on Field's statements
and one Hungarian translation of Field's "autobiography,"
all dated during the last two years of his confinement, convey
the same tale of relations with Hiss:
[Field and his wife] made friends with Alger Hiss
an official of the "New Deal" brought about by
Roosevelt and his wife. After a couple of meetings
we mutually realized we were Communists. Around the summer
of 1935 Alger Hiss tried to induce me to do service for
the Soviets. I was indiscreet enough to tell him he had
come too late. Naturally I didn't say a word about the Massings.
[Field and his wife] made friends with Alger Hiss
an official of the "New Deal" brought about by
Roosevelt and his wife. After a couple of meetings
we mutually realized we were Communists. Around the summer
of 1935 Alger Hiss tried to induce me to do service for
the Soviets. I was indiscreet enough to tell him he had
come too late. Naturally I didn't say a word about the Massings.
the same statements Field says Hede Massing was the Soviet
agent to whom Field turned over State Department documents
in the 1930s. The statements are consistent with Chambers'
and Massing's testimony. In two other prison "autobiographies"
Field refers to Hiss only as a colleague who knew that Field
"was a Communist." But in those statements, Field
goes on to note that Hiss, while aware that Field was a Communist,
was a strong supporter of Field at the State Department and
even tried to help him obtain a job as a State Department
adviser in the Philippines in 1940.
this would appear to be damaging, indeed devastating, new
evidence. It would make Field's anxiety over HUAC and those
letters to Hiss and Duggan far more suspicious. Perhaps Field
really did possess information that he feared HUAC could use
to "hurt my friends, especially Alger Hiss," as
he put it in one of those prison "autobiographies."
Perhaps the letters to Hiss and Field were communications
about strategy for keeping their cover. Indeed, the dossier
records a prison statement by Field that he briefly visited
Hiss in 1939 in America, where they agreed that if either's
cover was ever blown, he would communicate to the other indirectly.
Closer Look: "Lies as Truth"
a closer examination of the textual and historical evidence
of this "unimpeachable testimony" reveals grounds
for considerable skepticism about the "information"
in these prison statements. First, they are what police call
"raw investigative files." The files contain no
evidence that a Hungarian or other Soviet-bloc secret-police
officer ever attempted to confirm any of Field's statements
about Hiss.
more important, evidence suggests that Field's statements
were made under coercive circumstances and conditions of considerable
psychological distress.
a March 18, 1954 letter to the Communist Party's Central Committee
in Moscow requesting review of his case, Field attests that
he has been subject to "influential terrorist pressure
(from being beat up to being crippled til the starvation cure)"
[sic]. The letter goes on to explain that he is "physically
a coward" and his mistreatment has caused him to "confess
more and more lies as truth" until "finally I do
not only utter and write down the most horrible lies but partially
even believe them."
by individuals familiar with the Field case support this account
of brutality and coercion. "No Excuses," a
memoir published in Hungary in 1991 by Vladimir Farkas about
his own career as a Communist official and that of his father,
who oversaw Hungary's secret police and Stalinist show trials,
asserts that the Hungarians tortured Field horribly during
his interrogations.
person close to Field a survivor of Polish prisons
in the same period visited Noel and his wife four times
in Budapest after Noel's release. Although they avoided direct
discussions of their prison experiences, this individual recalls,
"Peripheral remarks that Noel [made] suggested that he
had had a pretty tough going. My feeling was that he was extensively
mistreated." Field's foster daughter, Erica Wallach,
when telling me about her own imprisonment in East Germany
and Russia, noted, "In prison I kept thinking, How
is Noel going to handle this kind of mistreatment?' He was
so sensitive and soft." Baczoni, speaking in his capacity
as a researcher familiar with interrogation records from this
era rather than in his capacity as director of the Interior
Ministry Historical Archive, confirms that political prisoners
at the time were routinely subject to "denial of food,
prolonged solitary confinement, denial of medicine, threats
to family members, beatings and psychological abuse."
legitimate grounds exist for concluding that the references
to Alger Hiss are among those statements coerced by the Hungarians.
All the prison references to Hiss have a seed prior to Field's
imprisonment. The recruitment story comes from Chambers's
HUAC testimony described in the aforementioned 1948 Hiss letter
received by Field. Moreover, Alger Hiss told me that he actually
did meet with Noel around 1939 in Washington, D.C. ("my
sole recollection is of seeing an old friend who had been
away for some time and doing good work"), and that he
did try to help Noel obtain a job in the Philippines in 1940.
Why would Field draw on this material to relate to his captors
stories about his relationship with Hiss, Soviet agent and/or
supporter of Communism, without regard for whether the stories
were necessarily true?
first explanation is straightforward. Anybody held in solitary
confinement for three years on charges of being an American
spy would try to think up any remotely believable stories
to relate to his accusers about how he was in fact connected
with supporters of Communism. The survivor of the Polish prison
told me how during internment he would go back to his cell
and "try to think up any believable Communist contact...
and then blow that up" in an effort to convince his captors
that he was a friend of Communism rather than an American
spy. Erica Wallach, imprisoned after she went to the Soviet
bloc looking for her foster father, says the same thing.
second explanation requires an understanding of the historical
context of the Fields' imprisonment. Field arrived in Budapest
during Hungary's "reign of terror," which Hungarian-American
historian Charles Gati recently concluded "turned out
to be much harsher than it was in the neighboring 'people's
democracies' of East-Central Europe and even the Soviet Union
itself." From 1948 to 1953, the Hungarian Communist Party
executed thousands, imprisoned tens of thousands and purged
approximately 200,000 from its own local ranks.
explanations for the purge focus on Field himself." In
1974 British investigative journalist Stewart Steven argued
in "Operation Splinter Factor" that a chief
source of the Stalinist purges in East-Central Europe was
a Western intelligence scheme masterminded by Allen Dulles
and centered on none other than Noel Field. Steven claims
to have uncovered information revealing that Dulles fed information
to double agent Jozef Swiatlo, a leader of Polish intelligence,
that was used to unleash a wave of paranoia throughout the
Soviet intelligence apparatus that the United States had infiltrated
Communist organizations throughout East-Central Europe via
"superspy" Noel Field. In fact, Field had the requisite
contacts with both the East and West necessary for contriving
such a tale. The alleged purpose of the Dulles scheme
Operation Splinter Factor was to drive the Communists
toward harsher tactics that would result in stunting the liberal-leaning
and nationalist elements in the East-Central European Soviet
satellites, thereby preventing Communism from attaining grass-roots
credibility there. According to Steven, Operation Splinter
Factor led to a decree from Lieut. Gen. Fedor Belkin, head
of the Southeast European Division of the Soviet Interior
Ministry, that there was a conspiracy, centered in Hungary,
of nationalists who, although Communist, were disloyal to
the Soviet Union. This type of suspicion in turn inspired
Hungary's paranoid terror campaign against Field and those
he may have known.
Lewis's 1965 book, "Red Pawn," cites a different
origin, but also identifies an important position for Noel
Field in the horrors in Hungary and the rest of East-Central
Europe. She hypothesizes that Stalin personally identified
Field as somebody with a portfolio of contacts with the dreaded
set of cosmopolitan Communists who had fought in the Spanish
Civil War, and therefore the dictator decided to use the myth
of Noel Field, "American master-spy," as a central
means to fulfill his desire to purge such figures from the
parties in the Soviet satellites.
the interrogations of Noel Field and his kin were used to
compile lists of hundreds of tainted Communists throughout
East-Central Europe, popularly known as "Fieldists,"
most of whom were either purged or executed. The tactics used
to create cases against the Fieldists appear to have been
similar to those techniques portrayed in Koestler's "Darkness
at Noon:" The interrogators would force the witness
to confess to a plausible detail about a contact and then
that detail would be distorted to create grounds for purging
the contact.
why would Field talk about his contacts with Alger Hiss? The
orders in the Field dossier indicate that the officials wanted
as many details as possible from Noel. Baczoni agrees: "The
general rule was, 'The more names mentioned the better.' This
rule applied in the Field case for sure." Laszlo Varga,
a part-time history professor who in 1991 became the first
freely elected director of the Budapest City Archive, explains,
"The secret police officers did not know what information
they wanted. The key was just to get as many names and details
of contacts as possible... to spin tales. Names meant power."
Professor Vera Pecsi, who recently wrote a documentary about
one of the many show trials in which the prosecutor created
a case for treason in part from alleged contacts with Noel
Field, agrees that the interrogators pressured Field to contrive
as many plausible stories about contacts with Communists as
Wallach, when discussing her own prison experience, told me
that she would intentionally create false stories about Communist
contacts in the West to appease her interrogators' demands
because she knew that those individuals were safe from the
Stalinist terror campaigns for which she was being used as
a source. This may be another reason that Noel Field had no
qualms about casting Hiss as a potential "Fieldist."
the Hungarian officials apparently cared little about whether
the details of contacts were truthful. Varga recalls a 1950
interrogation file in which a Hungarian poet names William
Shakespeare as one of his Communist contacts in the West.
Baczoni read a file in which a fearful witness offers an unbelievable
3,500 contacts. The secret police recorded such "information"
with no indication of skepticism.
Sealed Case?
conclusions about Alger Hiss may one draw from a critical
examination of the Noel Field dossier? All the documents written
under noncoercive circumstances such as the Field-Hiss
correspondence assume Hiss's innocence on their face,
while the apparently incriminating statements were made under
questionable, indeed brutal, circumstances.
most controversial archival documents about Hiss may have
yet to surface. A Russian source close to the K.G.B. archivists
told me that the K.G.B. does indeed have files on Hiss. But
these files, even if released, may only lead into another
labyrinth. Recent East-Central European experience reveals
the consistent inability of newly available Communist era
archives to "seal" history. In the former Czechoslovakia,
courts reviewing the "lustrated" (the Czech term
for those named in secret police files as collaborators) have
ended up reversing 90 percent of the cases because of the
ambiguity of the archival evidence. Similarly, Hungarian Prime
Minister Jozsef Antall's government over the past two years
has strategically leaked cryptic passages from archives to
slander rebels within Antall's ruling coalition. Thus far,
the Antall government's defamation-by-disclosure efforts have
largely backfired once the subjects have had the opportunity
to convey the full context of the misleading archival passages.
attempts to frame his articles as a response to this trend.
He criticizes last year's successful initiative by longtime
Hiss supporter John Lowenthal (on behalf of The Nation Institute's
Cold War Archives project) to have Col. Gen. Dmitry Volkogonov,
chairman of Russia's military intelligence archives, issue
a widely publicized declaration that the Soviet archives show
that the espionage allegations against Hiss are "completely
groundless" [see The Nation, "In Re Alger
Hiss:" November 16, 1992]. I disagree with Lowenthal's
strategy. But now Tanenhaus, ironically, employs the same
to Communist-era archives has provoked file fever on the left
and the right. The interpreters of newly available material
should remember that the secret-police interrogations were
"fantastic situations" and the convoluted mind games
that went on in those prison cells can only be understood
by "fantastic imaginations," as Arseny Roginsky,
a former Russian dissident and prisoner who now heads the
Memorial Society and served as an expert on archives to the
Russian Constitutional Court and Parliament, told me this
summer. Statements made in such a setting are neither "unimpeachable"
nor even "testimony," as those terms are used in
our legal tradition.
I left the Hungarian Interior Ministry Archive for the last
time, director Baczoni asked if I had found what I was looking
for. After I shrugged my shoulders, he chuckled. "Oh,
come on! Everyone leaves these archives with fantastic theories!"
Disclosures from secret-police archives, like those about
Hiss, provide good material for tabloids and talk shows, but
rarely "seal the case."
Klingsberg, an attorney, is the former executive director
of the Soros Foundation's Institute for Constitutionalism
and Legislative Policy. Research for this article was supported
by The Nation Institute's Cold War Archives project.


text:
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Hayden Lourd told us in his first solo scene that he loves dark haired, dark skinned guys and we immediately thought of Levi Karter. Physically, they make the perfect couple. Hayden's tall with blond hair and blue eyes, while Levi is a little shorter with brown hair and brown eyes. But it wasn't until we saw them hang out in person that we realized how adorable they actually are!
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- Be the first to Review
Your rating for Hollywood Walk of Fame
Expert Review of Hollywood Walk of Fame
Active and Adventure Review of Hollywood Walk of Fame
Active and AdventureExpert Tips & Helpful Hints
- Your favorite stars are identified by one of five icons in a bronze star: a theatrical mask, a radio microphone, a record, a motion-picture camera, and a television.
- The Walk of Fame spreads over five acres, lining both sides of Hollywood Boulevard from Gower to La Brea, and both sides of Vine Street, from Yucca to Sunset.
- The official groundbreaking of the Walk of Fame was in 1960, and since then more than 2,000 stars have been immortalized.
Family Review of Hollywood Walk of Fame
FamilyExpert Tips & Helpful Hints
- For kids, the best part of the Walk of Fame may be the characters near Grauman's Chinese Theatre. You could run into Superman, SpongeBob, and Captain Jack Sparrow all on the same stretch of sidewalk.
- There are about 2,400 stars—if you're looking for a specific person, you might want to map out his or her star in advance (or you may have a long walk ahead). Or take your iPhone: There's an app that will give you each star's location, information about the star, and more.
- New stars are born each year. Find out if one of your favorite actors or musicians (Russell Crowe, Adam Sandler, and Cameron Diaz are on the 2010 list) is due to be honored—maybe you'll catch him or her in person. The Hollywood Chamber of Commerce announces each ceremony a week in advance (www.hollywoodchamber.net).
7018 Hollywood Blvd.
Hollywood, California 90028
7018 Hollywood Blvd.
Hollywood, California 90028
Visitor Information
Telephone Number: (323) 469-8311
Website: http://www.hollywoodchamber.net
The details, dates, and prices mentioned here were accurate at the time of publication.
Review by Away Copyright © 2010 Away.com all rights reserved.
- Hollywood, CA Hotel Prices Made Easy
Orbitz.com - Just Orbitz and go!
- Hotel Deals in Hollywood, CA
Expedia.com - Don't just travel. Travel right!
- Book Now with Cheap Tickets and Save on Hollywood, CA Hotels!
CheapTickets.com - Just click. You're there.
- Find the best rates for hotels in Hollywood, CA
Kayak.com - Life's a trip.


text:
When: Wednesday, March 27, 2013 3:00 PM - 4:30 PM
Where: Dorchester Hall Basement Lounge
Event Type(s): Diversity,Meeting,Recreation
International Coffee Hour is a weekly opportunity for international students and scholars visiting the University of Maryland to come and meet the faculty, staff and students of the University. Rather than being exclusively for international visitors, it is a casual social gathering for a global interaction. Each week a different campus organization will sponsor the International Coffee Hour and provide free drinks and snacks for you to enjoy. See the schedule below for the list of sponsors.
Please join us anytime on Wednesdays between 3 and 4:30pm!
Website: http://www.international.umd.edu/ies/7490
For more information, contact:
Felixa Wingen
Office of International Services
+1 301 405 3385


text:
Quantity Available:
Since 1938, Cleveland-based Kichler Lighting has created exceptional lighting in a variety of styles, finishes, colors and designs. With a diverse collection of indoor and outdoor lighting in classic and contemporary styles, Kichler Lighting always focuses on making home lighting that is both beautiful and functional.
The Wilton Flushmount is available with the following:
- Antique Barley glass shade
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- Two 60 Watt 120 Volt Medium Base Incandescent lamps (not included).
- Fixture: Height 7", Diameter 20.5"
Manufacturer IDs: view
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text:
Tunes and training— published: 03-12-2014
Do you exercise to music? If you do, you might have noticed that you run faster when a fast-paced, upbeat tune comes on.
Exercise scientists noticed the relationship between exercise and music some time ago. For those of us who aren't trained athletes, music can have a huge effect on our performance and mood during exercise. Without realizing it, people push themselves harder during exercise when listening to fast-tempo music, which increases heart rate as well as speed, endurance, and in some cases the rate of perceived exertion. Exercisers also feel an improved sense of well-being when working out to music.
So why is it you prefer certain songs when you're exercising? One explanation suggests that a part of your brain tries to match the movement of your body to the beat of the music. In fact, scientists have found that when you listen to music with about 125–140 beats per minute, both your heartbeat and your movements synchronize to work at the most energy-efficient, optimal level for exercise. In essence, music works with your brain to coordinate your bodily functions and optimize your workout.
The best workout songs seem to share certain characteristics:
- 125-140 beats per minute during exercise; slower for warm-ups, cool-downs, and for some endurance-type exercises
- A motivational or upbeat message
- Familiar tunes or a preferred style of music
- A tempo that matches the rhythm of your exercise
Ask your buddies about their workout playlists too. They might have something totally different to offer—a new beat to stay fit with.
For more tips on how to optimize your workout, explore HPRC's Physical Fitness domain.


text:
Photos and Review by Oyster.com Investigators
On the outskirts of the Gothic Quarter, near the water in La Ribera
La Ribera is similar to the Gothic Quarter, with its gorgeous, centuries-old architecture and narrow streets. Stunning stone and wood buildings tower over skinny cobblestone streets lined with designer boutiques, tiny restaurants, and hole-in-the-wall cafes.
Stagnated for years, La Ribera (and El Born, its "neighborhood within a neighborhood") is now one of the hot spots in town, where locals mingle with fascinated tourists in bars and taverns housed in 18th-century buildings. El Mercat del Born, a beautiful glass and iron market built in the late 19th century, has some of the most incredible ruins in all of Barcelona, and 14th century Santa Maria del Mar Basilica is not to be missed.
The Pullman Barcelona Skipper is near the water, within walking distance of the beach and some sights. However, many of the top Barcelona sights are a 25 to 30 minute walk from the hotel, but the metro is close by for quicker access.
The Pullman Skipper is a contemporary hotel near Barcelona's beaches. Rooms are on the large side and have contemporary decor, floor-to-ceiling windows, and in some, terraces. Bathrooms are a standout feature, with large vanities, separate walk-in showers and soaking tubs, and towel warmers. The range of hotel features is impressive: two outdoor pools, including one on the rooftop, a small spa, and a variety of dining options. The lobby is airy and inviting and the rooftop bar has great views. While renting towels for the beach can be a bit of a nuisance, this hotel is near the metro for easy access to Barcelona's sights, and 24-hour room service is on offer.
Like Us on Facebook And Have the Best Vacation Ever


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Privacy and Cookies
Cessna is predicting a bright future for its business in Europe, despite the prevailing economic uncertainty on the continent.
Speaking this morning, Cessna president and chief executive Scott Ernest said: "We are seeing high levels of interest in our aircraft throughout Europe and we expect this trend to continue.
"The significant advantages that stem from business aviation are important and appealing factors in the European market. We expect the European Union to remain our single largest market outside the US for the foreseeable future."
Ernest said that alongside established markets such as the UK and Germany, it is seeing growth in eastern Europe for its jet models and "increasing demand" for its Pratt & Whitney Canada PT-6A-powered Grand Caravan in CIS countries.
It predicts its two in-development models - the midsize Citation Latitude and the light Citation M2 - will also find favour with European customers. Both models are attracting "encouraging levels of interest", the company says.
The $14.9 million Latitude is benefiting from a range extension announced in February, taking maximum range to 2,300nm (4,260km), it adds. It is scheduled to enter service in 2015.
Its sister aircraft, the Williams International FJ44-1AP-21-powered M2, made its first prototype flight, lasting 90min, in March this year. Since then, the prototype has flown 35 times and acquired more than 70h of flying time. Cessna also just completed the first flight of its second conforming prototype.
US Federal Aviation Administration certification is expected in the first half of 2013, followed by deliveries beginning in the second half of 2013.
Meanwhile, Cessna will open a second European Citation service centre in Valencia by the fourth quarter of 2012. The Valencia site offers 5,800m² (62,400ft²) of hangarage and 3,500m² of office and administration space.
Cessna is also positioning a mobile service unit in Stuttgart, Germany, with a small support office.
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All Stories
Newton Poll - 07/10/2010
"To the person who is complaining about why no one does anything for the western side of Newton County, why should we? It's mostly full of starter homes that don't pay enough in taxes to cover the services they use. It's something we can blame on the commissioners that wanted and want affordable housing here. That side of the county is DeKalb County the 3rd, Rockdale County is Jr. Thank goodness when Aaron Varner was chairman they made it mandatory for 2-acre lots in east Newton, which will hopefully keep the east side of Newton desirable by forcing the developers to build higher-priced homes. If you look at where all of the crime is occurring most of it is in west Newton. People may move here for a better life with intentions of being good citizens of the community, but it's your children, not the parents. No one owes them anything so they shouldn't act like it."
Teachers as students
Teachers from Newton and Rockdale counties attended a four-day professional development class this past week at the Rockdale Career Academy in Conyers.
Chamber holds final candidate forum
COVINGTON -- The final forum of the primary election season took place Thursday night.
Schools get CRCT results
COVINGTON -- School-level scores from the spring CRCT released Thursday afternoon show that many grade levels and schools performed at or below the state averages in several subject areas, while some performed above state averages.
20 from city choose early retirement
COVINGTON -- The city of Covington will eliminate up to 14 positions through a voluntary early retirement incentive package.
BOC OKs reduced-cost lawn contract
COVINGTON -- County commissioners approved a proposal by Durden's Lawn Maintenance Tuesday night to reduce its contract for a savings of more than $12,000.
School board candidate charged with simple battery
COVINGTON -- A candidate seeking election to the Newton County Board of Education is awaiting a legal decision on misdemeanor charges of simple battery stemming from a December incident at Liberty Middle School.


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Gates' gifts spur health efforts
focus on | global health
What do the French Family Science Center, DukeEngage and University HIV/AIDS research centers have in common?
Melinda Gates and global health.
Melinda French Gates, Trinity '86, Fuqua '87 and co-chair of the Bill and Melinda Gates Foundation, has retained an active influence on her alma mater's global health priorities since she left her position as a member of Duke's Board of Trustees in 2003.
Gates Foundation spokesperson Marie Groark said about 50 percent of the foundation's funds are allocated for reducing health inequities throughout the world, while the rest of the funding goes toward civil engagement and education programs.
Recent financial gifts from the Gates Foundation to Duke for the FFSC, DukeEngage and the Financial Aid Initiative resulted from conversations with the benefactors, President Richard Brodhead said.
"I communicate regularly with Melinda Gates and have met with her during visits to the Pacific Northwest. She is interested in Duke's priorities and, as with [former] President [Nan] Keohane, she has shared her thoughts on those priorities with me," Brodhead said.
Such priorities include achieving the goals set forth in the University's strategic plan, "Making a Difference," and furthering advances in global health initiatives.
The Gates Foundation and strategic priorities
Duke's new strategic plan "Making a Difference," approved in October 2006, aims to invest $1.3 billion above normal budgets throughout the next five to eight years in students, faculty, programming and facilities.
The top goals of the plan include attracting and retaining outstanding faculty, deepening undergraduate and graduate students' engagement in education, improving the campus with new and improved facilities, strengthening the arts and recommitting to diversity and access to education.
And the newly opened FFSC-which supports undergraduate laboratory education in biology and chemistry and faculty research in biology, chemistry and physics-will help achieve goals of the strategic plan, said John Simon, vice provost for academic affairs and George B. Geller professor of chemistry.
"The facility has been central to attracting outstanding faculty to Duke. One example of this is [Professor] Warren S. Warren in chemistry," Simon wrote in an e-mail. "The education laboratories in [the] Gross Chemistry [Building] were significantly out of date. The undergraduate labs in FFSC enable us to put forth the type of curricular offerings consistent with the aspirations in 'Making a Difference.'"
After it was originally designated the French Science Center in 2003, the Board of Trustees voted in 2006 to change the name in order to "acknowledge the ongoing support of the Bill and Melinda Gates Foundation and express to the Gates Family [the Board's] gratitude and appreciation."
The Gates Foundation donated $30 million for the building and $5 million for student life initiatives in 2002.
"Melinda French Gates is a wise and visionary leader at her alma mater. We're grateful not only for the resources provided by the Bill and Melinda Gates Foundation, but also for Melinda's personal leadership in helping us shape and implement university priorities," Keohane said when announcing the foundation's gift.
Groark said the Gates Foundation's scholarship donations, such as those for DukeEngage and the Financial Aid Initiative, span all three of the foundation's priority areas-global health, civic engagement and education.
"[The Gates Foundation is] building the next generation of leaders to gain an interest in and expertise in helping to solve these problems," Groark said.
Global health support at Duke
Funding for the Center for HIV/AIDS Vaccine Immunology falls specifically within the global health sector of the Gates Foundation's priorities, Groark said.
In 2003, CHAVI Director Dr. Barton Haynes and several other global health professionals, including Rick Klausner, then research director of the Gates Foundation, called for a new effort to speed the progress of a preventive AIDS vaccine.
"What followed was a process whereby the Gates Foundation made a partnership with the [National Institutes of Health] and they went together to the [international forum] G8 leaders and got their blessing for the effort," Haynes said.
The federal government committed $300 million, and the Gates Foundation put in $287 million for the effort. The U.S. grant was designated for the creation of CHAVI at Duke, and Haynes was named its principal investigator following a peer-reviewed competition.
At the same time, the Gates Foundation held a peer-reviewed grant competition for its $287 million, which went to support 16 HIV vaccine research centers worldwide as part of the foundation's Global HIV Vaccine Enterprise.
Duke received two of the centers-one for Haynes and one for professor of experimental surgery David Montefiori, co-director of the laboratory for immune measurements.
After a difficult application process, Haynes received the Gates Foundation grant in August 2006, he said.
"My Gates grant is to focus on figuring out how to induce the right kind of antibody to make a vaccine," Haynes said. "David Montefiori's center is a central laboratory to test everyone's vaccines that are developed."
In order to receive a grant from the Gates Foundation, an organization must submit a proposal, Groark said. Then the strategy of the proposal, as well as the leadership of the organization, is evaluated by the Gates Foundation.
Haynes said CHAVI plans to apply for funding for the clinical trials of the new vaccines within three years.
"[The next step is to] perform studies of our vaccine candidates in monkeys to determine if they make protective antibodies," he said.
The Global Health Institute, a partner of CHAVI, opened at Duke in November 2006.
Although the institute has not applied for funding from the Gates Foundation, GHI Director Dr. Michael Merson has a personal grant from the Gates Foundation that he brought with him to Duke from his former position at Yale University.
"[The Gates Foundation] tends to give support to projects in developing countries," Merson said. "They have a number of priority areas, such as HIV."
With his grant, Merson works with a project that aims to empower female sex workers in Andhra Pradesh, a southern state in India heavily impacted by HIV/AIDS.
"Sex workers are afraid to ask their partners to wear condoms because they are afraid they won't want to have sex with them," Merson said. "[The Gates-funded project] educates them to give them the knowledge and wisdom to insist their partners use a condom."


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Round Brush - Holbein, Number Three, circa 1993 Reg. No: SH 931262
- Summary:
- Alternative Name(s): Paint Brush
Number Three Round Brush made by Holbein, circa 1993. Used by artist Yoka Van Den Brink in Newport, Victoria, to paint items using Hindeloopen techniques. Hindeloopen craft techniques date back to the 16th century, to the port of Hindeloopen, in Friesland in the North of Holland where sailors developed a decorative tradition of handpainting domestic items to add colour to the dark houses through winter. Motifs include flowers, scrolls, buds, birds and sometimes bible scenes and landscapes.
- Description:
- Fine wooden paint brush with metal neck, fine bristles and covered in plastic protective sheath. The wooden handle is black in colour.
|18.50 cm (Height), 0.50 cm (Length)
More information
|Tagged with:
|cultural identity, cultural traditions, crafts, handcrafts, commerce, artistic practices, dutch communities, dutch immigration, hindeloopen art
|Themes this item is part of:
|Immigration & Artistic Practice Collection
|Primary Classification:
|Secondary Classification:
|Ethnicity - Creative Practice
|Tertiary Classification:
|On brush: 3 SERIES 655-GR HOLBEIN Japan
|Holbein, Japan, circa 1993
|Mrs Yoka Van Den Brink, Newport, Victoria, Australia, circa 1993
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