Abstract: 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.

Abstract: Granules are prepared containing an admixture of protein and salt layered over an inert particle. A preferred amount of salt is about between 63.7 and 84.3% of the total weight of the admixture. Proteins include pharmaceutically important proteins such as hormones, or industrially important proteins such as enzymes including proteases, amylases, lipases and cellulases capable of hydrolyzing substrates such as stains. Inert particles include inorganic salts, sugars, sugar alcohols, small organic molecules such as organic acids or salts, and minerals such as clays or silicates. A binder such as starch or polyethylene oxide may be mixed in with the admixture. A barrier material such as an inorganic salt or organic acid or salt may be in the admixture or coated over the admixture layer. A coating layer of a soluble or water dispersible film-forming polymer may be between the inert particle and admixture layer and/or over the admixture layer.

Abstract: An efficient method for therapeutic treatment of leukemia is provided in which a patient's body fluid during external circulation is brought into direct contact with an adsorbent material capable of specifically and selectively adsorbing leukemic cells in the body fluid. The leukemic cell-adsorbing material is a composite of a lectin protein coupled with a physiologically inert carrier material such as a galactan polysaccharide in the form of beads. The lectin protein may be obtained from a mushroom fungus such as Agrocybe cylindracea or a leguminous seed such as from the jequirity bean plant. The lectin protein and carrier material can be bound by forming chemical linkages between amino groups in the lectin protein and functional groups in the carrier material, and unreacted functional groups of the carrier material may be blocked with an amino acid. A leukemic cell-adsorbing column may be formed by filling the leukemic cell-adsorbing material into a tubular body to form an adsorbent bed.

Abstract: Granules are prepared containing an admixture of protein and starch layered over an inert particle. Proteins include pharmaceutically important proteins such as hormones, or industrially important proteins such as enzymes including proteases, amylases, lipases and cellulases capable of hydrolyzing substrates such as stains. Inert particles include inorganic salts, sugars, sugar alcohols, small organic molecules such as organic acids or salts, and minerals such as clays or silicates. The admixture may also contain sugar such as sucrose. A ratio of corn starch to sugar much greater than 1:1 such as in a range of about 5:1 to about 15:1 is preferred. A coating layer may be between the inert particle and the admixture and/or over the admixture. Methods that may be used in preparing the granules include pan-coating, fluid-bed coating, prilling, disc granulation, spray drying, extrusion, centrifugal extrusion, spheronization, drum granulation and high shear agglomeration.

Abstract: An immobilized microbial consortium is formulated which comprises of a synergistic mixture of all the bacterial strains of Aeromonas hydrophila, Pseudomonas fluorescens, Pseudomonas aeruginosa, Bacillus circulans, Yersinia enterocolitica, Enterobacter cloaca and Bacillus brevis. The formulated microbial consortium is immobilized on a non-biodegradable and economically cheaper support. The said immobilized microbial consortium is used for the biodegradation of synthetic phenol as well as phenol present in petroleum refinery effluent. The results of biodegradation obtained with the microbial consortium immobilized on coconut fiber are compared with those obtained with microbial consortium immobilized on well known support. The coconut fiber used for immobilization proved to be a better support than a well known support such as calcium alginate.

Abstract: A particle resistant to storage of at least one first and at least one second component, wherein

Abstract: Hydrocolloid gel beads containing enzymes immobilized therein are useful for catalyzing organic transformations in non-aqueous media. The enzyme is imbibed into a dehydrated hydrocolloid polymer gel bead. The resulting enzyme-laden bead may be dehydrated, if desired. A preferred hydrocolloid is carrageenan, especially kappa carrageenan. The resulting hydrocolloid gel beads are particularly adapted for catalyzing asymmetric transformations.

Abstract: An artificial pancreas is described herein which comprises one or more viable and physiologically active pancreatic islet cells capable of producing insulin, encapsulated within a semipermeable spheroidal membrane comprising agar gel. Further disclosed are a method for producing agar microbeads, a tissue implantation method and a reseeding method for the artificial pancreas.

Abstract: The present invention relates to a method for synthesizing optically active cyanohydrin. An immobilized enzyme is used in the invention, in which (S)-hydroxynitrile lyase is immobilized in a carrier comprising a porous inorganic material.

Abstract: This invention provides novel methods for the formation of biocompatible membranes around biological materials using photopolymerization of water soluble molecules. The membranes can be used as a covering to encapsulate biological materials or biomedical devices, as a “glue” to cause more than one biological substance to adhere together, or as carriers for biologically active species.

Abstract: The invention relates to a method of evaluating the immunological status of a subject comprising the steps of 1) determining the content of an antibody in a liquid sample from the subject using an immunoassay, wherein the reaction between the antibody of the sample and a ligand in the form of an antigen, an antibody or a hapten, the ligand being directed to the Fab region of the sample antibody, is carried out in the presence of other constituents of the sample to obtain a measurement 1, 2) determining the content of an antibody in the liquid sample using an immunoassay, wherein the reaction between the antibody of the sample and a ligand in the form of an antigen, an antibody or a hapten, the ligand being directed to the Fab region of the sample antibody, is carried out in the absence of other constituents of the sample to obtain a measurement 2, and 3) interrelating measurements 1 and 2 to express the interference and using the interference as a parameter for evaluating the immunological status of the subje

Abstract: Enzyme granules suitable for incorporating into detergents or cleaners are provided containing an enzyme, a carrier material and a granulation auxiliary containing phosphated starch. The phosphated starch preferably has a mean degree of phosphation ranging from 1.5 to 2.5. Carrier materials include starch, cereal flour, cellulose, alkali metal aluminosilicate, layer silicate and alkali metal salts. Enzymes include proteases, lipases, amylases and cellulases. A preferred carrier material contains water-swellable starch, sucrose, cereal flour and cellulose powder. The granulation auxiliary may contain a co-granulation auxiliary selected from polyethylene glycol having an average molecular weight of from 200 to 6,000, 1,2-propylene glycol and a poly-ethoxylate having a specified formula. Preferred granules have a mean particle size of from 0.3 to 3 mm.

Abstract: Cells in a matrix or in the matrix in a vessel are grown to form a multicellular aggregate. Pressure is exerted on the growing cells by the matrix or the matrix together with the vessel due to growing cellular mass displacing the matrix. A value representing pressure exerted on the cells is calculated and the pressure is modulated to control growth of the multicellular aggregate, or to produce a multicellular aggregate of a pre-selected size or a pre-selected size and shape. Matrices include agarose, alginate and collagen gels, and the pressure exerted on the cells can be non-isotropic. The cells may be tumor cells, or liver, pancreatic, brain, skin, bone or muscle cells, and cell growth can be in vitro or in vivo. When collagen forms the matrix, the matrix may contain glycosaminoglycan.

Abstract: Cells, preferably isolated pancreatic islet cells, are treated with a compound or a combination of compounds selected from an antioxidant, an anti-cytokine, an anti-endotoxin and an antibiotic. The compound or combination of compounds is in a medium for culturing the cells before microencapsulation, in a medium for cryopreserving the cells by freezing followed by thawing and microencapsulation, in a medium for culturing the cells after microencapsulation, or in a medium for culturing the cells before microencapsulation and in a medium for culturing the cells after microencapsulation. Microcapsules containing cells may be incubated with a physiologically acceptable salt to increase durability of the microcapsules. In a preferred method, isolated pancreatic islet cells are cultured for about 12 to about 36 hours in a medium containing an antioxidant as the compound, cryopreserved by freezing in a medium containing the compound or combination of compounds, thawed and microencapsulated.

Abstract: The present invention relates to the field of carbohydrate crosslinked glycoprotein crystals. Advantageously, such crosslinked glycoprotein crystals display stability to harsh environmental conditions, while maintaining the structural and functional integrity of the glycoprotein backbone. According to one embodiment, this invention relates to methods for concentrating proteins that have been modified by carbohydrates and for releasing their activity at controlled rates. This invention also provides methods for producing carbohydrate crosslinked glycoprotein crystals and methods for using them in pharmaceutical formulations, vaccines, immunotherapeutics, personal care compositions, including cosmetics, veterinary pharmaceutical compositions and vaccines, foods, feeds, diagnostics, cleaning agents, including detergents and decontamination formulations.

Abstract: A sterilization indicator for testing the effectiveness of a sterilization procedure comprises a source of an enzyme, a sterilant-resistant chemical associated with the enzyme, and a substrate that reacts with the enzyme to form a detectable enzyme-modified product that provides an indication of the failure of the sterilization procedure. The sterilant-resistant chemical may be a polyglycerol alkyl ester, polyglycerol alkyl ether, an ethoxylated polyhydric alcohol ester, or a polyhydric alcohol ether. The indicator may be used to test the effectiveness of a hydrogen peroxide plasma sterilization procedure and may be provided with a non-challenge test pack or a lumen-challenge test pack.

Abstract: Immune recognition of a transplant such as tissue implanted in a host mammal is obscured by encapsulating the transplant in a hydrogel matrix containing gelatin, dextran, at least one nitric oxide inhibitor and polar amino acids. The polar amino acids increase rigidity of the matrix and allow direct injection of the encapsulated transplant into a mammal without further immunosuppression. Preferably, the nitric oxide inhibitor is a combination of L-cysteine and an L-arginine analogue such as aminoguanidine, and the polar amino acids are a combination of L-glutamic acid, L-lysine and L-arginine. The matrix may also contain a superoxide inhibitor such as EDTA. Implanting can be carried out by applying a buffer medium containing a nitric oxide inhibitor to an implant site, implanting the encapsulated transplant, and applying to the implant site a buffer medium which may contain a nitric oxide inhibitor. The buffer medium may also contain a superoxide inhibitor.

Abstract: Methods and reagents are provided for specifically targeting biologically active compounds such as antiviral and antimicrobial drugs, or prodrugs containing the biologically active compound to specific sites such as specific organelles in phagocytic mammalian cells. The biologically active compound or prodrug is linked to a microparticle with a linker that is non-specifically or specifically cleaved inside a phagocytic mammalian cell. Alternatively, the biologically active compound or prodrug is impregnated into a porous microparticle or coated on a nonporous microparticle, and then coated with a coating material that is non-specifically or specifically degraded inside a phagocytic mammalian cell. The prodrug contains the biologically active compound linked to a polar lipid such as ceramide with a specific linker such as a peptide that is specifically cleaved to activate the prodrug in a phagocytic mammalian cell infected with a microorganism.

Abstract: Methods for providing a pathogen-free pig or pig fetus as a donor of tissue, cells and/or organs to a recipient human. Animals are free of zoonotic pathogens. When fetal tissues are used for transplantation, donor animals are free from zoonotic pathogens, pathogens able to cross the placental barrier, and tissue-specific pathogens, e.g., neurotropic pathogens. Tissues, cells and organs from pigs free of the above-listed pathogens are suitable for transplantation into humans, include fetal neuronal cells for treatment of Parkinson's disease and islet cells for treatment of islet insufficiency-related diseases.

Abstract: Substances such as chemical substances and biological substances including animal, vegetable and microbial cells are encapsulated using a process and an apparatus wherein a coil through which alternating current flows causes a magnet to vibrate creating vibrations such as in the range of between 300 to 4000 Hz that are transmitted to an encapsulating fluid containing the substance to form small substantially spherical particles containing the substance. The apparatus includes a pulsation chamber containing a movable wall for receiving the encapsulating fluid containing the substance to be encapsulated. A nozzle is spaced downstream from the pulsation chamber for receiving the encapsulating fluid from the pulsation chamber. A permanent magnet is mounted on the movable wall, and a coil is spaced from the permanent magnet by an air gap and is located proximate to the permanent magnet.

Abstract: Cells, preferably isolated pancreatic islet cells, are treated with a compound or a combination of compounds selected from an antioxidant, an anti-cytokine, an anti-endotoxin and an antibiotic. The compound or combination of compounds is in a medium for culturing the cells before microencapsulation, in a medium for cryopreserving the cells by freezing followed by thawing and microencapsulation, in a medium for culturing the cells after microencapsulation, or in a medium for culturing the cells before microencapsulation and in a medium for culturing the cells after microencapsulation. In a preferred method, isolated pancreatic islet cells are cultured for about 12 to about 36 hours in a medium containing an antioxidant as the compound, cryopreserved by freezing in a medium containing the compound or combination of compounds, thawed and microencapsulated. The microcapsule contains a hydrogel core such gelled alginate and a semipermeable outer membrane such as formed with polylysine.

Abstract: A device is prepared having cells or tissue attached to a non-degradable filamentous matrix surrounded by a semi-permeable membrane. The matrix is preferably formed of a plurality of monofilaments twisted into a yarn or woven into a mesh, and can be in the form of a cylinder. When implanting the device, the semi-permeable membrane is preferably immunolsolatory, and the cells or tissue may produce a biologically active molecule to provide therapy. To enhance cell or tissue adhesion, the matrix is coated with extracellular matrix molecules or treated to provide a surface charge. The device can be made by inserting the matrix into a capsule formed of the semi-permeable membrane, distributing the cells or tissue on the matrix through an opening of the capsule, and sealing the opening of the capsule.

Abstract: A material for suppressing proliferation of cancer cells is produced by entrapping cancer cells in a selectively-permeable structure such as a bead, and culturing the entrapped cells in a culture medium. Entrapment restricts growth of the cancel cells during culturing and causes the cells to produce in the culture medium a material having a molecular weight of at least about 30 kd as measured by filtration that suppresses proliferation of cancer cells. The material is separated from the culture medium by filtering the medium through a filter that separates material having a molecular weight of at least about 30 kd from material having a molecular weight of less than 30 kd. The structure that entraps the cells may contain 10,000 to 500,000 cells. The material or the structure containing the entrapped cells that produce the material can be administered to a subject to suppress cancer cell proliferation.

Abstract: A succinimide polymer is produced by thermally polymerizing an ammonium salt of aspartic acid in the presence of an acid catalyst such as a boric acid catalyst. Another amino acid may be added for copolymerizing with the ammonium salt of aspartic acid. Ammonia liberated during the production of a succinimide polymer can be collected in a fumaric acid suspension, an acidic fumaric acid solution, a maleic acid solution or an acidic maleic acid solution, and the resultant liquid reacted with an enzyme which may be immobilized to produce L-aspartic acid. An aspartic acid polymer is produced by hydrolyzing the succinimide polymer with a basic substance.

Abstract: A system for nitrate removal from aquariums, both fresh water and marine aquariums, by means of permeable polymeric beads which contain a combination of fermentative and denitrifying bacteria and a carbon source. Preferred beads are beads made from sodium alginate or chitosan. The bacteria, in the presence of the carbon source, are able to reduce nitrate to nitrogen gas. Bacteria which are not harmful to fish are used. The porous beads used in the process are novel and part of the invention.

Abstract: Activated support materials are provided containing oxirane or azlactone groups as substituents in linear polymers as activated groups. A base support containing hydroxyl groups is suspended in a solution containing cerium (IV) ions and a monomer containing an oxirane or azlactone group, and grafting polymerization is carrier out to produce a polymer containing oxirane or azlactone groups covalently bonded to the base support. Azlactone groups can be bonded to the base support via a thioether bond by using a base support containing thiol groups. The activated support materials can be used to prepare affinity supports containing an affinity ligand that is thiophilic or possesses a metal chelating group, or to prepare immobilized enzymes. The ligand can be iminodiacetic acid, or can be obtained by reacting an oxirane group of the support material with NaHS, and reacting the resultant product with divinylsulfone followed by reacting with mercaptoethanol.

Abstract: A device that includes a living cell or tissue and an agent that inhibits the ability of a host molecule to damage the cell or tissue. The device can be constructed in various forms including an implantable device, a composite microreactor and a double composite microreactor. The composite microreactor includes an internal particle that includes a living cell or tissue, an internal particle matrix that includes the living cell or tissue and an internal semipermeable coating enclosing the internal particle matrix, a gel super matrix in which the internal particle is embedded, and an agent that inhibits the ability of a host molecule to damage the cell or tissue. The double composite microreactor includes an internal particle, a particle that includes a particle matrix in which the internal particle is embedded, a super matrix in which the particle is embedded, and an agent that inhibits the ability of a host molecule to damage the living cell or tissue.

Abstract: Process for the preparation of a &bgr;-lactam antibiotic in which a &bgr;-lactam nucleus is subjected to an enzymatic acylation reaction with the aid of an acylation agent at a molar ratio of acylation agent/&bgr;-lactam nucleus of less than 2.5, with the acylation agent and/or the &bgr;-lactam nucleus being supersaturated in the reaction mixture during at least part of the acylation reaction. In the process, a concentrated slurry or solution, for instance, of the &bgr;-lactam nucleus and/or the acylation agent with a different pH or a higher temperature than the pH or temperature at which the acylation reaction is carried out is added to the reaction mixture during the acylation reaction. Both the &bgr;-lactam nucleus and the acylation agent may be supersaturated in the reaction mixture.

Abstract: Gel immobilized enzymes are prepared for use in carrying out reactions in hydrophobic solvents. The gel is formed from a gelatinizer such as gelatin or a polysaccharide such as agarose, agar, pectin, sodium alginate or carrageenan. The gel contains a ratio of amount of gelatinizer to amount of water such that the gel is capable of being mechanically divisible into dimensionally substantially stable fragments at a temperature which may reach a lower limit of the gelation temperature range of the gelatinizer. A preferred gel contains 30 to 50% gelatin, and is prepared by forming a mixture of water, enzyme and water-soluble gelatin, heating the mixture to dissolve the gelatin to form a solution, and cooling the solution until it forms a gel.

Abstract: Enzymes are immobilized for use in non-aqueous enzymatic reactions by dehydrating hydrocolloid gel beads having an average particle size of 5 to 150 microns and a network structure capable of swelling in aqueous media, immersing or suspending the dehyrated gel beads in an aqueous solution of enzyme where the beads swell and imbibe the enzyme solution, optionally dehydrating the resultant enzyme-containing gel beads and recovering the gel beads containing the enzyme. In a specific method, the hydrocolloid is carrageenan such a kappa carragennan, the enzyme is subtilisin Carlsberg, the aqueous enzyme solution contains 0.05% to 40 wt % enzyme and the amount of enzyme imbibed is 0.05 to 0.5 grams of enzyme per gram of dehydrated gel beads.

Abstract: Lipase is immobilized on surfaces to facilitate oil removal from the surfaces and to alter wettability of the surfaces. The lipase is isolatable from a Pseudomonas organism such as Pseudomonas putida ATCC 53552 or from an organism expressing a coding region found in or cloned from the Pseudomonas. A particularly preferred lipase has a molecular weight of about 30 to 35 kd and is resolvable as a single band by SDS gel electrophoresis. Lipase sorbed on fabric forms a fabric-lipase complex for oil stain removal. The lipase may be sorbed on fabric before or after an oil stain, and the lipase is active to hydrolyze an oil stain on dry fabric or fabric in laundering solutions. The sorbed lipase has enhanced stability to denaturation by surfactants and to heat deactivation, is resistant to removal from fabric during laundering, retains substantial activity after drying fabric at an elevated temperature, and retains activity during fabric storage or wear.

Abstract: The present invention relates to the field of carbohydrate crosslinked glycoprotein crystals. Advantageously, such crosslinked glycoprotein crystals display stability to harsh environmental conditions, while maintaining the structural and functional integrity of the glycoprotein backbone. According to one embodiment, this invention relates to methods for concentrating proteins that have been modified by carbohydrates and for releasing their activity at controlled rates. This invention also provides methods for producing carbohydrate crosslinked glycoprotein crystals and methods for using them in pharmaceutical formulations, vaccines, immunotherapeutics, personal care compositions, including cosmetics, veterinary pharmaceutical compositions and vaccines, foods, feeds, diagnostics, cleaning agents, including detergents and decontamination formulations.

Abstract: Natural product metabolites produced by fungi are synthesized using fungal spores that have been immobilized onto/into a support. Supports that can be used include loofah sponge, synthetic sponge, powdered cellulose paper, wood shavings, calcium alginate gel beads, agar gel beads, channeled aluminum beads, polypropylene beads and glass beads. Immobilizing the fungal spores provides accelerated production of the natural product metabolite in a standard bioreactor. Fungi that can be used include Penicillium cyclopium, Penicillium chrysogenum, Penicillium citrinum, Trichoderma viride, Aspergillus terreus and Monascus ruber. In a preferred embodiment, spores of Penicillium cyclopium NRRL 6233 immobilized with loofah sponge are cultured in a nutrient media for at least 4 days to produce compactin which is a hypocholesteremic agent.

Abstract: This invention provides novel methods for the formation of biocompatible membranes around biological materials using photopolymerization of water soluble molecules. The membranes can be used as a covering to encapsulate biological materials or biomedical devices, as a “glue” to cause more than one biological substance to adhere together, or as carriers for biologically active species. Several methods for forming these membranes are provided. Each of these methods utilizes a polymerization system containing water-soluble macromers, species which are at once polymers and macromolecules capable of further polymerization. The macromers are polymerized using a photoinitiator (such as a dye), optionally a cocatalyst, optionally an accelerator, and radiation in the form of visible or long wavelength UV light. The reaction occurs either by suspension polymerization or by interfacial polymerization.

Abstract: Immobilization of SH group-containing compounds on a solvent-insoluble support is carried out in the presence of an antioxidant to prevent oxidation of SH groups to S—S bonds. This improves immobilization efficiency and suppresses deterioration of inherent characteristics of the SH group-containing compound. Antioxidants include sodium pyrosulfite (sodium disulfite), sodium sulfite, sodium hydrogensulfite, sodium hydrosulfite and L-ascorbic acid. SH group-containing compounds include cysteine, peptides or proteins containing cysteine and thiol compounds such as ethanethiol, aminoethanethiol, benzylthiol and thiophenol. Preferably, the SH group-containing compound has a molecular weight not more than 3×104. The support may be activated by a functional group such as glycidyl, imidocarbonato, tosyl, tresyl, carboxyl, amino, azido or hydroxyl. The support can be inorganic such as glass beads or organic such as a synthetic polymer or a polysaccharide.

Abstract: A method of determining the presence of chronic volume dependent hypertension is provided wherein a determination is made as to whether there has been a substantial reduction in phosphorylation of the blood-derived protein or renal proximal brush border membrane protein and if such reduction exists concluding that chronic volume dependent hypertension exists in a patient. The method may advantageously be practiced by employing blood serum or blood plasma as the body specimen containing the protein in determining whether a patient has chronic volume dependent hypertension, a cellular component of the blood, such as a blood-derived protein coming from the plasma membrane of lymphocytes. The method may include subsequent therapeutic patient treatment. Related diagnostic apparatus is also provided.

Abstract: Microparticles such as propagules of eukaryotic biocontrol agents are encapsulated in cellular-scale polymer capsules that have a diameter similar to normal eukaryotic cells in a range of about 10 &mgr;m to about 400 &mgr;m. The microparticles are encapsulated by adding a hydrophobic dispersion medium such as a mixture of chloroform and hexane or a mixture of corn oil and n-hexadecane having a specific gravity of about 1 and containing an emulsifier such as lecithin to an aqueous suspension of the microparticles and a polymer matrix precursor such as alginate, agitating vigorously to form a stable emulsion of microscopic globules containing a microparticle, and adding the emulsion to an aqueous solution containing a polymerizing agent such as calcium chloride to polymerize and precipitate the globules to form microparticles encapsulated in polymer matrix capsules that may be of a teardrop shape having a length of 40-200% longer than the diameter.

Abstract: The invention relates to water-soluble polymeric thiosulfates, to a method for their preparation by polymer-analogous addition of tetrathionate to unsaturated polymers and to their application in surface coating.

Abstract: Oxalate-degrading enzymes and bacteria were encapsulated for both enteric and intraperitoneal administration. We have shown that via alginate microencapsulation of Oxalobacter formigenes, enzymatic activity was retained for several months. A new process was developed which strengthened the alginate microcapsules and their tolerance to citrate treatment. Much smaller (30-50 &mgr;m) alginate microcapsules were made for injection as means of implantation. For oral administration, multi-encapsulated microspheres of cellulose acetate phthalate in poly-2-vinylpyridine (pKa=3.5) were prepared to protect the enzymes from gastric juices.

Abstract: A material for suppressing proliferation of cancer cells is produced by entrapping cancer cells in a selectively-permeable structure such as a bead, and culturing the entrapped cells in a culture medium. Entrapment restricts growth of the cancel cells during culturing and causes the cells to produce in the culture medium a material having a molecular weight of at least about 30 kd that suppresses proliferation of cancer cells. The material is separated from the culture medium by filtering the medium through a filter that separates material having a molecular weight of at least about 30 kd from material having a molecular weight of less than 30 kd. The structure that entraps the cells may contain 10,000 to 500,000 cells.

Abstract: A size modified fibrinolytic enzyme, wherein the size of the enzyme is modified by covalent attachment of at least one large organic molecule to the enzyme.

Abstract: A method of producing a three dimensional section of polymerized liquid. The method includes providing a molding apparatus which includes a mold having at least one side, at least one piece of porous material adjacent to the one side of the mold and at least one support layer adjacent to the piece of porous material. The molding apparatus is clamped together and a liquid is added between the frame and the porous material. The filled molding apparatus is placed in a bath of polymerizing agent until the liquid polymerizes to form a three dimensional section.

Abstract: A method of producing a dehydrated hydrogel comprises dispensing fibers into an aqueous solution of a hydrogel precursor material incorporating a plasticiser, the fibers incorporating cations which are capable of cross-linking said precursor material to form a hydrogel, and freeze drying the mixture thus produced to provide a dehydrated hydrogel which incorporates said fibers, the dehydrated hydrogel being cross-linked by said cations.

Abstract: Microbial spores having increased sensitivity to sterilants are provided. An additive such as a dipeptide, oligosaccharide, and/or polyhydroxyalcohols are added to the spores wherein the additive is bound to sterilant-sensitive sites in the spores. The additive increases sensitivity of the spores to a sterilant. More than one additive can be utilized to alter the sensitivity of the spores to a sterilant. Biological indicators comprising the microbial spores and a solid support are also disclosed and those spores having a dipeptide specifically bound to sterilant-sensitive sites in the spores have an altered sensitivity to a sterilant. Furthermore, a method is disclosed for altering the sensitivity of microbial spores to a sterilant comprising drying the spores at a temperature between 35° C. and 55° C. in a liquid composition having an amount of the additive therein.

Abstract: Isolated chondrocytes are propagated in the presence of a biological gel such as a fibrin gel to generate a cartilage matrix that firmly bonds together two adjacent cartilage pieces. A bonding composition containing the isolated chondrocytes mixed with the biological gel is applied to a surface of one (or both) of the cartilage pieces, and the surface is contacted with the other cartilage piece. In a different order of steps, the two cartilage pieces are held in apposition, and gaps at the interface are filled with the bonding composition. In another method, either or both of the cartilage pieces are first incubated with the isolated chondrocytes, the biological gel is then applied, and the cartilage pieces are held together. Alternatively, after incubating with isolated chondrocytes, the biological gel can be applied to fill gaps at the interface between cartilage pieces held in apposition.

Abstract: A compound having a polysaccharide binding domain such as contained by a cellulose and essentially lacking in polysaccharidase activity is purified from other ingredients in a mixture using an affinity partition system. A mixture containing the compound is contacted with a system containing as a first phase an aqueous solution of oligosaccharide polymer such as cellulose and as a second phase a solution of a polymer such as a poly(ethylene glycol)-poly(propylene glycol) copolymer. The compound petitions into the first phase and binds to the oligosaccharide polymer, preferably with a Ka of 103 to 107, to form a complex. The complex is collected, and the compound is dissociated from the oligosaccharide polymer. The compound may be formed of a non-peptide chemical moiety or a peptide moiety linked to a polypeptide having the polysaccharide binding domain.

Abstract: An amphiphilic enzyme is immobilized by preparing an emulsion containing a continuous hydrophobic phase and a dispersed aqueous phase containing the enzyme and a carrier for the enzyme, and removing water from the dispersed phase until this phase turns into solid enzyme coated particles. The enzyme is preferably a lipase, and the immobilized lipase can be used for reactions catalyzed by lipase such as interesterification of mono-, di- or triglycerides, de-acidification of a triglyceride oil, or removal of phospholipids from a triglyceride oil when the lipase is a phospholipase. The aqueous phase may contain a fermentation liquid, an edible triglyceride oil may be the hydrophobic phase, and carriers include sugars, starch, dextran, water soluble cellulose derivatives and fermentation residues. A substance to be processed such as triglycerides, diglycerides, monoglycerides, glycerol, phospholipids or fatty acids may be in the hydrophobic phase.

Abstract: A bioartificial extracellular matrix for use in tissue regeneration or replacement is provided by derivatizing a three-dimensional hydrogel matrix with a cell adhesive extracellular matrix protein or cell adhesive peptide fragment of the protein. Preferably, derivatizing is by covalent immobilization of a cell adhesive peptide fragment having the amino acid sequence, ArgGlyAsp, TyrIleGlySerArg or IleLysValAlaVal. Cartilage or tendon can be regenerated by implanting a matrix containing an adhesive peptide fragment that favors chondrocyte invasion. The matrix can be pre-seeded with cells, and tissue can be reconstituted in vitro and then implanted. A cell-seeded matrix can be encapsulated in a semi-permeable membrane to form a bioartificial organ. An agarose hydrogel matrix having an agarose concentration of 0.5-1.25% (w/v) and an average pore radius between 120 nm and 290 nm is preferred.

Abstract: An adsorbent for use in direct hemoperfusion to adsorb and remove harmful substances from blood is prepared by immobilizing a sulfated polysaccharide and/or its salt on a water-insoluble carrier. Preferably, the sulfated polysaccharide has a limiting viscosity of 0.005 to 0.5 dl/g and a sulfur content of 5 to 22% by weight, and is immobilized on the carrier in an amount of 0.02 to 200 mg per ml of carrier. Carrier particles can have an average particle size of 30 to 5000 .mu.m, and preferably 120 to 800 .mu.m. The sulfated polysaccharide inhibits adhesion of hemocytes and exhibits an anticoagulation property to extend blood coagulation time, and has additional functions of adsorbing releasing factors released from hemocytes, adsorbing lipoproteins, and enabling reduction of carrier particle size to about 30 .mu.m. Immobilizing a ligand on the carrier with the sulfated polysaccharide makes it possible to adsorb and remove specific substances in blood that bind to the ligand.

Abstract: Microorganisms or enzymes immobilized in beads are prepared using a combination of calcium alginate, polyethylene glycol (PEG) and polyethylene imide (PEI). An aqueous solution containing 1-14% by weight each of calcium alginate, PEG and PEI is combined with a concentrated solution of a microorganism or an enzyme to form a mixture. The mixture is combined with an aqueous solution of 4-8% w/v CaCl.sub.2 to form spherical beads that are allowed to remain in the solution for 3-4 hours. Thereafter, the beads are rinsed with water for 2-3 minutes, transferred to water in a mixer, and stirred with a magnet for 6-9 hours. The resultant beads containing the microorganism or enzyme can be used for removing inorganic nitrogen and organic carbon in waste water, or in processes for making biochemical products.