Abstract: A lawn assay is described for determining compounds that affect enzyme activity or that bind to target molecules. Compounds to be screened are cleaved, and diffused from solid supports into a colloidal matrix. Enzymatic catalysis or binding to target molecules by the compounds is carried out in the matrix. Active compounds are found by monitoring a photometrically detectable change in a substrate, coenzyme, or cofactor involved in the enzymatic reaction, or in a labeled ligand bound to the target molecule, that produces a zone of activity associated with the compounds.

Abstract: A structurally stable gel bead containing an entrapped enzyme and a method for its manufacture. The enzyme is covalently cross-linked to gelatin in the presence of glutaraldehyde prior to the formation of the gel bead, to prevent leakage of the enzyme. Propylene glycol alginate is then added to the mixture. Once the gel beads are formed, they are then soaked in glutaraldehyde, which imparts structural stability to the gel beads. This method can be used with many types of enzymes, such as proteases, carbohydrases, proteases, ligases, isomerases, oxidoreductases, and specialty enzymes. These and other enzymes can be immobilized in the gel beads and utilized in a number of enzymatic processes. Exogenously added ions are not required to maintain the structural stability of these gel beads.

Abstract: Water soluble macromers are modified by addition of free radical polymerizable groups, such as those containing a carbon-carbon double or triple bond, which can be polymerized under mild conditions to encapsulate tissues, cells, or biologically active materials. The polymeric materials are particularly useful as tissue adhesives, coatings for tissue lumens including blood vessels, coatings for cells such as islets of Langerhans, coatings, plugs, supports or substrates for contact with biological materials such as the body, and as drug delivery devices for biologically active molecules.

Abstract: Methods and compositions are provided for controlling cell distribution within an implantable bioartificial organ by exposing the cells to a treatment that inhibits cell proliferation, promotes cell differentiation, or affects cell attachment to a growth surface within the bioartificial organ. Such treatments include (1) genetically manipulating cells, (2) exposing the cells to a proliferation-inhibiting compound or a differentiation-inducing compound or removing the cells from exposure to a proliferation-stimulating compound or a differentiation-inhibiting compound; exposing the cells to irradiation, and (3) modifying a growth surface of the bioartificial organ with extracellular matrix molecules, molecules affecting cell proliferation or adhesion, or an inert scaffold, or a combination thereof. These treatments may be used in combination.

Abstract: Water soluble macromers are modified by addition of free radical polymerizable groups, such as those containing a carbon-carbon double or triple bond, which can be polymerized under mild conditions to encapsulate tissues, cells, or biologically active materials. The polymeric materials are particularly useful as tissue adhesives, coatings for tissue lumens including blood vessels, coatings for cells such as islets of Langerhans, coatings, plugs, supports or substrates for contact with biological materials such as the body, and as drug delivery devices for biologically active molecules.

Abstract: A bioartificial three-dimensional hydrogel extracellular matrix derivatized with a cell adhesive peptide fragment is provided for use in tissue regeneration or replacement. The choice of adhesive peptide fragment depends on the desired target cell type. Cartilage or tendon can be regenerated by implanting a matrix containing adhesive peptide fragments that favor 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 gel pore radius between 120 nm and 290 nm is preferred. The peptide fragment preferably contains the sequence, ArgGlyAsp or TyrIleGlySerArg or IleLysValAlaVal, and is covalently immobilized to the matrix.

Abstract: The present invention provides methods and a device for producing minimal volume capsules containing viable cells or cellular aggregates. The methods and device use a two-phase aqueous emulsion system to form a dispersion of liquid capsule-forming materials in a continuous liquid phase to which is added a suspension of biological material. Alternatively, the biological material can be added to one or the other of the liquid phases. The composition of this emulsion is adjusted to promote the thermodynamically-driven process of particle engulfment by the dispersed droplets of liquid capsule-forming materials. Subsequently, the droplets engulf the biological material to form a liquid film surrounding the tissue and are converted to solid form, resulting in encapsulation of the biological material in minimum volume capsules.

Abstract: Decarbamylases are provided capable of producing D-.alpha.-amino acids by hydrolysis of N-carbamyl-D-.alpha.-amino acids. A source of the decarbamylases is recombinant microorganisms produced by gene manipulation methods. Decarbamylases having improved thermostability can be obtained in which amino acids at a thermostability-related site of a natural decarbamylase have been replaced with other amino acids by mutating a DNA fragment encoding the natural decarbamylase. Recombinant DNA is obtained from a vector DNA and a DNA fragment encoding a natural decarbamylase where the nucleic acid sequence encoding an amino acid at a thermostability-related site is replaced with a nucleic acid sequence encoding another amino acid. The recombinant DNA is used to produce transformants that produce thermostable decarbamylases.

Abstract: This invention is related to a process for the enhanced immobilization of a ligand to solid supports to be used for a biochemical detection method. The enhanced immobilization of the ligand was obtained by coating the surface of the solid support with the adhesive polyphenolic protein isolated from mussels. The bound ligand is reacted with a solution containing an antiligand whereby the antiligand becomes bound to the immobilized ligand. After removing the excess or unbound antiligand, the antiligand bound to the immobilized ligand is detected by using an enzyme-linked immunoassay.

Abstract: A method is provided for producing a sterile and pyrogen-free column containing coupled protein for use in removing a predetermined substance from the blood of a human subject. The method abrogates sterilization of the finished protein-containing product by providing sterile and pyrogen-free raw materials at each production step. The method provides a pathogen-free, purified solution of protein which binds to a predetermined substance in human blood such as LDL or immunoglobulin. Typically, the protein is anti-human LDL immunoglobulin or anti-human Ig immunoglobulin. The method also provides a sterile and pyrogen-free column matrix material such as an agarose which is chemically activated, either using CNBr and triethylamine or using 1,1'-carbonyldiimidazole.

Abstract: Substantially dust-free enzyme-containing particles are prepared by spray-coating solid particulate cores with a fermentation broth containing enzymes. Fermentation broths containing from 5% to 100% total enzyme solids and 4%-55% total solids content in the broth may be used. For broths containing 4%-31% total solids, enzyme and coating agents may be spray-coated so as to achieve a weight gain of 1%-24% w/w over the initial weight of the cores. For broths with 31%-55% total solids, a corresponding weight gain of 1%-100% may be achieved. The enzyme coated cores may be further coated with coating agents. The total dry weight added to the cores ranges from 1% to 19% in the absence of coating agents and 1% to 24% in the presence of coating agents when the lower range of total solids content is used. The total dry weight added to the cores ranges from 1% to 100% when the higher range of total solids content is used.

Abstract: A matrix structure containing attached cells such as endocrine cells, fibroblasts, endothelial cells or genitourinary cells is implanted in a patient adjacent tissue having a high surface area and vasculature such as mesentery, omentum or peritoneum tissue. Large volumes of cells can be attached to the matrix and the matrix implanted with minimum trauma and blood loss into a patient to produce a functional organ equivalent. Multiple matrix structures containing cells can be implanted to functionally resemble naturally occurring organs. Implanting multiple matrices between folds of the mesentery is particularly well suited for growth of endocrine structures, including liver, pancreas, and adrenal gland. The matrix structure is preferably formed from a biodegradable artificial polymer. Collagen and non-biodegradable materials can also be used, and the matrix structure can be overlaid with a material that enhances cell attachment.

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: Methods and compositions are provided for controlling cell distribution within a bioartificial organ by exposing the cells to a treatment that inhibits cell proliferation, promotes cell differentiation, or affects cell attachment to a growth surface within the bioartificial organ. Such treatments include (1) genetically manipulating cells, (2) exposing the cells to a proliferation-inhibiting compound or a differentiation-inducing compound or removing the cells from exposure to a proliferation-stimulating compound or a differentiation-inhibiting compound; exposing the cells to irradiation, and (3) modifying a growth surface of the bioartificial organ with extracellular matrix molecules, molecules affecting cell proliferation or adhesion, or an inert scaffold, or a combination thereof. These treatments may be used in combination.

Abstract: A core material such as animal tissue or cells is contained within a semipermeable vessel which may be a microcapsule, hollow fiber or plastic membrane having a semipermeable wall by a method that prevents the core material from incorporation into the wall of the vessel. This is accomplished by suspending the core material in a solution of polysaccharide gum such as an alkali metal alginate in an amount between about 0.2% and about 0.5%, removing and washing the core material to remove all but a thin layer of polysaccharide gum, and gelling the polysaccharide gum with multivalent cations or other means to form a pretreated core material.

Abstract: A fungal inoculum for bioaugmentation of soils contaminated with hazardous compounds or spawn for use in the edible mushroom industry is disclosed. A mechanically pelleted substrate that contains both structural and nutritive components forms the core of the fungal inoculum. The pelleted substrate core is coated with a hydrophilic material in which fungal propagules are dispersed. The biological potential of the fungal inocula can be enhanced by formulating the material composition of fungal inocula to meet the specific requirements of a particular fungus.

Abstract: The disclosed invention is a device for adhering cells in a specific and predetermined position. The device comprises a plate defining a surface and a plurality of cytophilic islands that adhere cells which are isolated by cytophobic regions to which cells do not adhere and further is contiguous with the cytophilic islands. The islands or the regions or both may be formed of a self-assembled monolayer (SAM). Further, the cytophobic regions are wide enough such that less than 10 percent of the cells adhered to the cytophilic islands are allowed to form bridges across the cytophobic regions and contact each other. The device is used in a method for culturing cells on a surface or in a medium and also for performing cytometry. Furthermore, the device is used in immobilization of cells at a surface and for controlling the shape of a cell.

Abstract: Fibers of a biocompatible, biodegradable or non-biodegradable, synthetic polymer are provided, and are formed into a three-dimensional scaffold. The fibers of the scaffold may have a branched configuration extending outwardly from a central stem. The fibers provide sufficient surface area to permit attachment to the scaffold in vitro of an amount of cells effective to produce functional vascularized organ tissue in vivo. Fibers of the scaffold are spaced apart such that the maximum distance over which diffusion of nutrients and gases must occur through a mass of cells attached to the fibers is between 200 and 300 microns. The diffusion provides free exchange of nutrients, gases and waste to and from cells proliferating throughout the scaffold in an amount effective to maintain cell viability throughout the scaffold in the absence of vascularization.

Abstract: A cell-scaffold composition is prepared in vitro for implanting to produce functional organ tissue in vivo. The scaffold is three-dimensional and is composed of hollow or solid fibers of a biocompatible, synthetic polymer which is biodegradable or non-biodegradable. The fibers of the scaffold may have a branched configuration extending outwardly from a central stem. Fibers of the scaffold are spaced apart such that the maximum distance over which diffusion of nutrients and gases must occur through a mass of cells attached to the fibers is between 200 and 300 microns. The diffusion provides free exchange of nutrients, gases and waste to and from cells proliferating throughout the scaffold in an amount effective to maintain cell viability throughout the scaffold in the absence of vascularization.

Abstract: A method is provided for immobilization of whole microbial cells containing enzymes in Ca-alginate capsules. Whole microbial cells are mixed with a CaCl.sub.2 solution, a small amount of xanthan gum is added, the resultant mixture is added drop-wise while stirring to a Na-alginate solution containing a small amount of surfactant which is preferably polyoxyethylenesorbitan monolaurate to obtain the capsules containing the cells. The capsules may be washed with distilled water, hardened by in a CaCl.sub.2 solution and incubated in a growth medium. The microbial cells are bacteria or fungi cells and may be recombinant cells.

Abstract: A method for grafting a cell in the brain of a mammalian subject is accomplished by attaching the cell to a support matrix so that the cell attaches to the matrix surface, and implanting the support matrix with the attached cell into the brain. A syringe containing viable cells that are attached to a matrix surface may be used to transplant the cells into the brain or spinal cord of a mammalian subject. Preferred support matrices are glass or plastic microbeads, either solid or porous, having a diameter from about 90 to about 125 .mu.m. The method employs cells of different types, preferably cells of neural or paraneural origin, such as adrenal chromaffin cells. Also useful are cell lines grown in vitro. Cells not of neural or paraneural origin, such as fibroblasts, may also be used following genetic alteration to express a desired neural product such as a neurotransmitter or a neuronal growth factor.

Abstract: Microspheres, of controllable shape and size, encapsulating active ingredients, are made by the internally controlled gelation of an emulsion including a water-soluble polysaccharide, a salt of a di- or trivalent metal cation, a polymerization inhibitor, water, a water-immiscible solvent (as a non-aqueous phase), and the active ingredient. The components of the aqueous phase, containing the water-soluble polysaccharide, polymerization inhibitor, di- or trivalent metal salt, active substance, and water, are blended together. This aqueous phase is then gradually mixed with the oil phase and agitated to form an emulsion. After sufficient time for solidification, the emulsion is broken and the resulting microspheres are collected. The active ingredient may be various substances, including live microorganisms.

Abstract: Methods and artificial matrices for the growth and implantation of cartilaginous structures and surfaces and bone are disclosed. In the preferred embodiments, chondrocytes are grown on biodegradable, biocompatible fibrous polymeric matrices. Optionally, the cells are proliferated in vitro until an adequate cell volume and density has developed for the cells to survive and proliferate in vivo. One advantage of the matrices is that they can be cast or molded into a desired shape, on an individual basis, so that the final product closely resembles a patient's own ear or nose. Alternatively, flexible matrices can be used which can be manipulated at the time of implantation, as in a joint, followed by remodeling through cell growth and proliferation in vivo. The cultured cells can also be maintained on the matrix in a nutrient media for production of bioactive molecules such as angiogenesis inhibiting factor.

Abstract: A biodegradable particulate vector for transporting biologically active molecules is prepared containing a nucleus for containing a biologically active molecule, a first layer of fatty acid compounds covalently bonded to the nucleus and a second layer of phospholipids hydrophobically bonded to the first layer. The nucleus is between 10 nm and 10 .mu.m in size and is formed of a cross-linked polysaccharide or oligosaccharide matrix onto which ionic ligands are uniformly grafted. The cross-linked polysaccharide may be dextran, cellulose or starch cross-linked with epichlorohydrin. The ligand may be an acidic compound selected from succinic acid, phosphoric acid, citric acid, glycine, alanine, glutamic acid and aspartic acid, or a basic compound such as choline, hydroxycholine, 2-(dimethylamino)ethanol or 2-(dimethylamino)ethylamine fastened onto the matrix via the acidic compound. The polysaccharide or oligosaccharide may be co-cross-linked with a protein such as keratin/collagen or elastase.

Abstract: Compositions containing two species of indolyl-3-alkane alpha-hydroxylase (INDH) are isolated from Pseudomonas XA. An INDH1 composition contains protein subunits having molecular weights of 75,000, 34,500 and 32,500 daltons. An INDH2 composition contains protein subunits having molecular weights of 60,000, 44,000 and 42,000 daltons. Molecular weights are determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The compositions have a Specific INDH Activity of at least 10 international Units of INDH activity per milligram of protein, and contain less than 1 nanogram of endotoxin per International Unit of Specific INDH Activity. The INDH compositions may be immobilized on an insoluble matrix such as silica beads to provide at least 2.5 international Units of INDH activity per gram of Immobilized INDH composition. The INDH compositions are isolated by lysing Pseudomonas XA cells at a temperature of no more than 15.degree. C.

Abstract: The invention relates to a method of detection, a sensor and a test-kit which find application in immunological detection (e.g., immunoassay). The invention provides, inter alia, a method of detection, suitable for use in immunological detection of an entity, which method includes the use of a secondary species (as defined in the specification), the use of a first detectable species, and the use of a second detectable species. The method may include, for example, the use of a primary species, a secondary species, a first detectable species and a second detectable species. The primary species may be, for example, an antibody or a ligand. The secondary species may be, for example, an auxiliary species such as an auxiliary binder or an auxiliary ligand, or a species which has a part which is an auxiliary function. The entity to be detected may be an analyte species as such or may be an entity which carries or includes analytes species.

Abstract: A cell-polymeric solution is injected into an animal where the polymer crosslinks to form a polymeric hydrogel containing dispersed cells and the cells form new tissue in the animal. The polymer is biodegradable and is a natural polymer such as alginate or a synthetic polymer. The cells are chondrocytes, osteoblasts, muscle cells, fibroblasts or cells acting primarily to synthesize, secret or metabolize materials. Crosslinking of the polymer results from using cations or anions, altering the pH or changing the temperature. A polyion such as polyethyleneimine or polylysine can be added before injection to stabilize the polymeric hydrogel. A kit for tissue formation is provided by combining the cell-polymeric solution with a means for injecting the solution into an animal.

Abstract: An implantable artificial endocrine pancreas comprising a reactive body of soft, plastic, biocompatible, porous hydratable material supporting a multiplicity of endocrine pancreatic islets in isolated spaced relationship from each other; and, a microporous barrier membrane enveloping and supporting the body, in spaced relationship from the pancreatic islets therein and through which molecules having a molecular weight greater than 60,000 Daltons cannot move.

Abstract: A biomass which can be used directly as panification ferment without previous separation of the culture medium and the biomass and without addition of industrial yeast is prepared by cultivating at least one strain of yeast in a culture medium specified in the description. Preferably, the said ferment comprises as yeast the strain Saccharomyces cerevisiae steineri DSM 9211.

Abstract: A biomass constituted of yeast and of lactic acid bacteria, which can be used directly as panification ferment without previous separation of the culture medium and the biomass, is prepared by cocultivating at least one strain of yeast and at least one strain of lactic acid bacteria in a mixed and/or sequential culture, in a culture medium specified in the description. Preferably, the said ferment comprises as yeast the strain Saccharomyces cerevisiae steineri DSM 9211 and as lactic acid bacteria one or several strains of Lactobacillus brevis DSM 9209, Lactobacillus plantarum DSM 9208, Leuconostoc mesenteroides DSM 9207 and/or Pediococcus pentosaceus DSM 9210.

Abstract: A transplant with a core of a viable, physiologically active, cell(s) and a non-fibrogenic coating of alkaline earth metal alginate having a high mannuronate to guluronate molar ratio and free from fibrogenic amounts of fucose, sulfate, phloroglucinol and protein moieties. The coating has a permeability sufficiently low and a thickness sufficiently large to protect the tissue cells from host immunological agents after transplantation, the coating also being sufficiently permeable and thin to permit the diffusion of cell sufficient nutrients and cell products through the coating required for cell viability. The alginate coating can be reacted with polylysine to form a polylysine-alginate complex on the outer surface thereof. The complex can then be reacted with polyaspartic acid to provide a physiologically acceptable negative surface charge.

Abstract: An electrostatic process is used for coating a biological material with a uniform, continuous polymer layer by discharging a suspension of the biological material in a gelable coating polymer solution in a continuous stream through an orifice into an electrostatic field. The stream is attenuated to form droplets by maintaining an electrostatic voltage between the needle and the gelling solution which is sufficient to maintain an attraction of at least 1.times.10.sup.-6 newtons on the stream of liquid leaving the needle, and the droplets are collected in a gelling solution. A preferred product is pancreatic islets having a continuous, smooth coating of high polymannuronate non-fibrogenic alginate having a thickness less than 200 .mu.m such as about 20-200 .mu.m. The alginate preferably contains less than 1 wt. % fucose, less than 0.5 wt. % sulfate and less than 0.01 wt. % phloroglucinol, is free of fibrinoogenic concentration of protein, and has a mannuronate to guluronate ratio of from 1.2 to 6.

Abstract: A supported polyionic hydrogel containing biologically active material is epared by impregnating into a porous support material a solution of anionic polysaccharide, a solution of cationic polysaccharide and a biologically active material. The anionic polysaccharide and the cationic polysaccharide react with each other to form a hydrogel containing the biologically active material. The hydrogel may be dried such as by lyophilization. The biologically active material can be in either polysaccharide solution or in another solution impregnated into the porous support material. A paper material or a textile material may be used as the porous support material. Preferably, the anionic polysaccharide is xanthan, dicarboxystarch or dicarboxycellulose and the cationic polysaccharide is chitosan. Especially preferred is a hydrogel formed from xanthan and chitosan.

Abstract: The present invention provides an immobilization medium which can immobilize bioreagents to support materials and which dries to a water resistant layer or film. The immobilization medium comprises (i) a liquid or fluid binding reagent and (ii) complexes of a bioreagent immobilized to a solid phase which are evenly dispersed within the binding reagent. The suspension can further include supplemental ingredients evenly dispersed throughout the medium which can provide the medium with electrochemical properties, enhance the stability of the immobilized bioreagent and/or improve the medium's capability of drying to a substantially water resistant or insoluble layer. The immobilization medium provided by the instant invention is in the form of a homogeneous liquid suspension.

Abstract: Elongated seamless capsules containing biological material are prepared by a method in which a coagulant, which includes a cell suspension or other biological material, and a polymeric casting solution are extruded through a common extrusion port having at least two concentric bores, such that the coagulant is extruded through an inner bore and the polymeric casting solution is extruded through an outer bore. Extrusion of the coagulant is initiated subsequent to initiating delivery of the casting solution to form a capsule having a curved and smooth leading edge shape. Delivery of the coagulant is then shut off, and extrusion of the casting solution is terminated either immediately or after some predetermined time. This procedure can be modified to form in the capsule a coaxial rod that is connected to one end but not the other end of the capsule.

Abstract: Biological agents such as secretory cells are encapsulated in a hydrophilic gel made of agarose or collagen-agarose and gelatin sponge-agarose combinations. In a preferred embodiment, semi-solid beads are formed from a suspension containing collagen, agarose and secretory cells such as pancreatic islets, the collagen is polymerized to form solid, agarose-collagen beads and the solid beads are coated with agarose. Coating is preferably by rolling the solid beads in about 5-10% agarose, contacting the rolled beads with mineral oil and washing oil from the beads. Beads containing secretory cells can be transplanted into a mammal to treat a condition caused by impaired secretory cell function.

Abstract: A coating apparatus includes a rotary cup for forming beads and projecting them radially outwardly, and one or more collection basins surrounding the bead forming cup. The cup is adjustably rotatable about its central axis, and the collection basins are independently rotatable and positioned to collect the beads projected from the cup. The coating apparatus further includes an elevation adjustment system for axially adjusting the alignment of the cup with respect to the selected collection basins. The rotational speeds of the cup and the collection basins are selected so as to minimize the impact of the beads against a gelling solution in the collection basins. In use, a supply mixture is introduced into a mixing chamber of the cup. As the cup spins, the coated particles are propelled upwardly by the centrifugal force from the mixing chamber along the inner surface of the cup, and are projected radially outwardly, as beads, into the gelling solution in one of the selected basins.

Abstract: A process and apparatus are provided for making solid Particles from an ionically cross-linkable material by cross-linking drops of the material with a cross-linking agent in the form of a falling stream. In one embodiment, a stream of the cross-linking agent flows down the inner walls of an enclosure and drops of the material are directed to the stream of cross-linking agent. In another embodiment, a stream of the cross-linking agent is free-falling by gravity in a cascade without contacting any surface and drops of the material are directed to the stream of cross-linking agent. Solid particles are separated from the cross-linking agent at about the bottom of the enclosure or at about the bottom of the cascade. The drops of cross-linkable material are directed at the stream of cross-linking agent preferably at an angle of incidence of less than 90.degree. such as between 5.degree. and 45.degree. and most preferably between 15.degree. and 30.degree.. Particles having a size of 10 .mu.

Abstract: A polysaccharide gel enclosing microorganisms is soaked in a solution of a high concentration such as at least 500 g/l of hydrophilic substance and the gel is at least partially dehydrated to provide improved viability of the microorganisms after storage and rehydration of the gel. The dehydration may be carried out in a fluidized bed or by lyophilization. The gel may be in the form of beads or fibers having a double layer structure formed by an internal layer or core of gel containing the microorganisms and an external layer or envelope of gel essentially devoid of the microorganisms. The hydrophilic substance can be a low molecular weight polyol such as glycerol or a sugar such as sucrose, glucose or fructose. The microorganisms in the gel are preferably yeasts and after rehydration the yeast-containing gel is used in the secondary fermentation of wine to produce sparkling wine or champagne.

Abstract: A polysaccharide gel enclosing microorganisms is soaked in a solution of a high concentration such as at least 500 g/l of hydrophilic substance and the gel is at least partially dehydrated to provide improved viability of the microorganisms after storage and rehydration of the gel. The dehydration may be carried out in a fluidized bed or by lyophilization. The gel may be in the form of beads or fibers having a double layer structure formed by an internal layer or core of gel containing the microorganisms and an external layer or envelope of gel essentially devoid of the microorganisms. The hydrophilic substance can be a low molecular weight polyol such as glycerol or a sugar such as sucrose, glucose or frutose. The microorganisms in the gel are preferably yeasts and after rehydration the yeast-containing gel is used in the secondary fermentation of wine to produce sparkling wine or champagne.

Abstract: A biocompatible microcapsule containing living cells encapsulated in a membrane is disclosed. The membrane is a complex formed by the cohesion of two polymer layers. An inner layer comprises a substrate biopolymer and an outer layer comprises a synthetic polyelectrolyte having an electrolytic charge opposite that of the substrate biopolymer. Droplets of a solution of substrate biopolymer containing a suspension of living cells can be added to a solution comprising the synthetic polyelectrolyte to form the encapsulates. The membrane is formed by the cohesion of the oppositely-charge polymer layers to form a complex of substrate biopolymer and synthetic polyelectrolyte. Preferably, the inner layer contains a cationic biopolymer, such as collagen modified to have a pKI of 9, or an anionic biopolymer such as esterified or modified hyaluronic acid.

Abstract: A solid support for immobilization of microorganism cells is made of a synthetic polymer such as a polyolefin, in admixture with an organic polymeric plant material such as corn fibers, oat hulls, starch, and cellulose. Preferably, the synthetic polymer is in an amount of about 50-95% wt-% and the plant material is in an amount of about 5-50 wt-%. Preferred polyolefins are polyethylene and polypropylene. The plant material may be a mixture including a plant material that functions as a nutrient to enhance growth of the microorganism on the support. The support may be produced by combining the synthetic polymer and plant material to form a composite, dough-like thermoplastic composition. The composition may be prepared in an extrusion mixer and co-extruded as an extrudate to form a shaped article.

Abstract: A multilayer analysis element for determination of total cholesterol is prepared having a light-transmissive water-impermeable support, at least one hydrophilic polymer layer on said support and a spreading layer on said hydrophilic polymer layer(s), and containing in one or more of the layers:(a) at least one enzyme having cholesterol esterase activity,(b) cholesterol oxidase,(c) peroxidase,(d) a color reagent composition, which in the presence of hydrogen peroxide and the peroxidase, produces a color change(e) at least one bile acid compound selected from the group consisting of bile acids, bile acid derivatives and salts of said acids and derivatives, and(f) an alkyl phenoxy polyethoxy ethanol containing in the alkyl group 1 to 20, preferably 7 to 10 carbon atoms, and a polyoxyethylene chain composed of at least 16, preferably 30 to 60 oxyethylene units.

Abstract: A method for preparing liquid-core microcapsules for cell cultures, using a hardening solution containing CaCl.sub.2 and polyethyleneimine to harden gel-core beads before coating them with polylysine solution.

Abstract: A method for multiple layer coating of biological tissue and cells for transplantation. The cell or tissue transplants are coated with multiple coatings of purified alginate. The method includes applying the first coat of sodium alginate gelled with divalent cations followed by optional treatment with strontium, barium or other divalent cation, resuspending the single coated droplets in sodium alginate and forming the halo layer around the first coating via exchange or diffusion of divalent cations from the single coating to the surrounding soluble alginate, removing the excess coating and gelling the remaining thin layer of soluble alginate with divalent cations. The coated transplants have distinct structure where biological tissue or cell core is covered with the first alginate coat, which is surrounded by an intermediate halo layer which is covered by the outer coating.

Abstract: In accordance with the present invention, there are provided methods to render cells non-adhesive and/or non-immunogenic with respect to macromolecules typically encountered in culture media or in physiological media. The invention method comprises contacting cells with an effective amount of a composition comprising a polycationic species having water-soluble polymer chains grafted thereon.

Abstract: An ionically gellable material is gelled with a metal cation and the metal cation content of the gel is reduced to provide the gel with binding sites not occupied by the metal cation so the gel can be used to bind and remove metal cations from solution. In a preferred embodiment, a calcium alginate or calcium pectate gel in the form of beads is prepared, the calcium ion content of the gel is reduced to between 0.01 mg/g and 1.5 mg/g of moist gel by contacting the gel with an aqueous solution of acid such as lactic or tartaric acid having a pH of 1 to 3.5. The gel can be produced containing a microorganism such as yeast used for fermentation so metal ions can be removed while fermenting with the microorganism. In the bottle fermentation of wine to produce champagne, the gel containing yeast is added to the wine in the bottle. During fermentation, calcium and potassium ions are bound by the gel to reduce the precipitation of calcium tartrate and/or potassium bitartrate.

Abstract: A reagent semi-sphere is disclosed comprising at least one biological reagent and a glass forming filler material in a concentration sufficient to facilitate formation of a glassy, porous composition, wherein the reagent semi-sphere is room temperature stable, water soluble, and has a T.sub.g above room temperature.

Abstract: A chemical specie is immobilized in a three dimensional, crosslinked matrix by bringing together in covalent bonding proximity a desired chemical specie and a polymeric coupling compound such as a photoderivatized polymer having at least two latent photochemical reactive groups per molecule, each latent reactive group being capable when activated of covalently bonding to another coupling compound molecule or to the chemical specie. The chemical specie may be a protein, carbohydrate, nucleic acid or lipid, and desirably is free of latent reactive groups that are activated upon activation of the latent reactive groups of the coupling compound. The latent reactive groups are simultaneously activated to cause formation via covalent bonding of a three-dimensional molecular network in which molecules of the chemical specie are covalently bonded to molecules of the coupling compound, and molecules of the coupling compound are covalently bonded to each other.

Abstract: This invention relates to a method of directing a therapeutic agent to selected cells, wherein a complex is formed between a polysaccharide capable of interacting with a cell receptor and a therapeutic agent. The resulting complex is administered to a subject, and permitted to be internalized into the selected cells through a process known as receptor mediated endocytosis (RME). The polysaccharide may be, for example, arabinogalactan, gum arabic, mannan or hydrolysis products thereof; the therapeutic agent may be, for example, an antiviral agent, a nucleic acid, hormone, steroid, antibody, chemoprotective or radioprotective agent. The cell receptor may be for example, the asialoglycoprotein receptor or the mannose receptor.