Abstract: Described are compositions with tethered growth effector molecules, and methods of using these compositions for growing cells and tissues. Growth effector molecules, including growth factors and extracellular matrix molecules, are flexibly tethered to a solid substrate. The compositions can be used either in vitro or in vivo to grow cells and tissues. By tethering the growth factors, they will not diffuse away from the desired location. By making the attachment flexible, the growth effector molecules can more naturally bind to cell surface receptors. A significant feature of these compositions and methods is that they enhance the biological response to the growth factors. The method also offers other advantages over the traditional methods, in which growth factors are delivered in soluble form: (1) the growth factor is localized to a desired target cell population; (2) significantly less growth factor is needed to exert a biologic response.

Abstract: An electrochemical sensor system and membrane and method thereof for increased accuracy and effective life of electrochemical and enzyme sensors.

Abstract: A method for manufacturing polyhydroxyalkanoate-containing structure, at least a part of a base material surface of the structure being coated with polyhydroxyalkanoate, the method comprises the steps of immobilizing a polyhydroxyalkanoate synthase on the base material surface, synthesizing, on the base material surface, polyhydroxyalkanoate using a 3-hydroxyacyl coenzyme A to become the substrate of the synthase and the synthase and coating at least a part of the base material surface with the synthesized polyhydroxyalkanoate, wherein the synthase contains an amino acid sequence capable of binding to the base material.

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: Encapsulated producer cells which are capable of expressing a molecule which is an inhibitor of CNS tumour growth provide a novel approach to the treatment of tumours, such as brain tumors which are localized within the central nervous system.

Abstract: A multi-layered microcapsule has an inner extracellular matrix and an outer shell. The inner extracellular matrix includes a first inner layer of biopolymer and a second intermediate layer of polymer that provides partial immune-protection and holds the first layer in place. The outer shell can form an exoskeleton to provide mechanical stability. Each of the individual layers can be varied to optimize mechanical stability, cell function, and immuno-protection.

Abstract: The heated support can be manufactured by subjecting an immobilized microorganism support in which activated sludge is immobilized by entrapment to heat treatment. Thus, it is possible to support particular microorganisms at high concentrations in an immobilizing material without pure culturing of the microorganisms.

Abstract: A non-disruptive three-dimensional culture system allows cell growth and proliferation in three dimensions, permitting cell splitting without subjecting cells to disruptive conditions that affect cell structure and functions. An extracellular matrix provides a good environment for culturing or co-culturing anchorage-dependent cells. The cells cultured this manner can be readily used in such applications as cell transplantation, tissue engineering seeding of cells on scaffolds, and other applications that require immediate availability of functioning cells.

Abstract: The present invention provides a novel silk-fiber-based matrix having a wire-rope geometry for use in producing a ligament or tendon, particularly an anterior cruciate ligament, ex vivo for implantation into a recipient in need thereof. The invention further provides the novel silk-fiber-based matrix which is seeded with pluripotent cells that proliferate and differentiate on the matrix to form a ligament or tendon ex vivo. Also disclosed is a bioengineered ligament comprising the silk-fiber-based matrix seeded with pluripotent cells that proliferate and differentiate on the matrix to form the ligament or tendon. A method for producing a ligament or tendon ex vivo comprising the novel silk-fiber-based matrix is also disclosed.

Abstract: A separation procedure for separating a selected desired or undesired population from a biological sample utilizing relatively heavy, dense particles and gravity sedimentation. The particles have one or more reactants bound thereto which are specific to and will bind with the selected population. The particles preferably are mixed with the sample by repeatedly causing the particles to settle through a substantial portion of the sample to bind to the selected population. The particles with the bound selected population then are allowed to preferentially settle in the sample and the supernatant including an enriched population is separated from the particles with the selected population bound thereto. The enriched populations in the biological sample supernatant can be further enriched by multiple removal steps.

Abstract: A method of making a silicone rubber having a structure adapted for growth of cells or living tissue, which comprises contacting a silicone rubber precursor with a biologically-acceptable sacrificial filler, curing the resultant mixture and removing the sacrificial filler to form a structured silicone rubber. The sacrificial filler is preferably an inorganic salt that has been ground, and the salt is selected from metal halides, metal carbonates and metal bicarbonates.

Abstract: A new human chitinase having an amino acid sequence as shown in FIG. 1 or FIG. 2. Modified forms of it having a similar chitin-hydrolyzing activity, and antigenic peptides representing one of its epitopes. Recombinant production of the human chitinase by genetically engineered hosts or host cells. Recombinant nucleic acid encoding it, and human chitinase-specific oligonucleotides. Use for therapeutic or prophylactic treatment of humans against infection by chitin-containing pathogens, or for decomposing chitin, e.g. from chitin-based articles. Antibodies binding to the human chitinase. Diagnostic test kits comprising the human chitinase, its antigenic peptides, human chitinase antibodies, recombinant nucleic acid or oligonucleotides.

Abstract: The present invention provides a masking system for selectively applying cells to predetermined regions of a surface. A mask is positioned adjacent to a surface to cover some portions of the surface while allowing other portions of the surface to remain uncovered. Cells then are applied to uncovered portions of the surface and the mask removed. Alternatively, a cell-adhesion promoter is applied to uncovered portions of the surface, and then cells are applied to the surface before or after removal of the mask from the surface. The masking system can be pre-coated, at least on those surfaces which will come into contact with cells, with a cell-adhesion inhibitor to resist absorption of cells and thereby avoid cell damage when the mask is removed (if cells are deposited prior to removal of the mask). A polymeric elastomeric mask that comes into cohesive-conformal contact with a surface to be patterned can be used.

Abstract: The present invention relates to human Akt3 proteins and polypeptides. The invention also relates to isolated nucleic acids encoding human Akt3, to vectors containing them and to their therapeutic uses, in particular for gene therapy. Expression of Akt3 inhibits cell death associated with hypoxia, apoptosis or necrosis.

Abstract: Methods and compositions are described that provide scaffolds for the support of cells. The scaffolds of the present invention have structural uniformity and desirable mechanical properties that make them suitable for a variety of uses, including uses for in vitro tissue regeneration or in vivo tissue or organ replacement. A method is described for controlling three-dimensional structure of the hydrogel/cell constructs under tissue culture environment.

Abstract: Hydrogels containing catalase co-immobilized with an analyte-sensitive enzyme such as glucose oxidase are disclosed. The hydrogels may be pH-sensitive, and preferably are thin and lightly crosslinked. The catalase is present in concentrations ranging generally from 100 units/ml to about 1000 units/ml. These hydrogels have much faster swelling response times as compared to hydrogels without catalase, and are useful in biosensors and analyte-responsive drug delivery devices. The hydrogels also have an increased useful life, due to protection of the immobilize analyte-sensitive enzyme from degradation by hydrogen peroxide.

Abstract: Compositions of matter are described which contain restricted cancer cells. When so restricted, the cells produce an unexpectedly high amount of material which suppresses cancer cell proliferation. The phenomenon crosses cancer type and species lines. Processes for making these compositions, and their use, are also described.

Abstract: The sensitivity of enzyme-based polarographic electrodes to oxygen concentration can be significantly reduced or eliminated by providing an oxygen-reservoir in intimate contact with the oxidative enzyme. This is achieved by making a stabilized emulsion between the enzyme and a compound in which oxygen is extremely soluble. An aqueous glucose oxidase solution is emulsified with a perfluorocarbon liquid, and the resulting emulsion is stabilized by chemically crosslinking the mixture to form a gel. Thin layers of the emulsion are fabricated by spreading a layer of the liquid emulsion before gelation occurs. Additional carrier proteins such as albumin may be added to the enzyme prior to crosslinking to protect enzymatic activity and enhance gel strength. Additional electron transport compounds may be added to further reduce sensitivity to oxygen concentration.

Abstract: This invention provides methods of retentate chromatography for resolving analytes in a sample. The methods involve adsorbing the analytes to a substrate under a plurality of different selectivity conditions, and detecting the analytes retained on the substrate by desorption spectrometry. The methods are useful in biology and medicine, including clinical diagnostics and drug discovery.

Abstract: This invention relates to a bioprocess engineering solution for a product removal process for use in a biofermentation. The invention discloses a process for withdrawing an aliquot of broth from a biofermentation vessel during at least a portion of the biofermentation, removing biocatalyst and water, chromatographically separating biofermentation products from the withdrawn broth using water as an eluent, and returning the remaining components of the broth back to the biofermentation vessel. Process chromatography permits highly selective separation of the target molecule, preventing feedback inhibition of the biofermentation.

Abstract: Compositions of matter are described which contain restricted cancer cells. When so restricted, the cells produce an unexpectedly high amount of material which suppresses cancer cell proliferation. The phenomenon crosses cancer type and species lines. Processes for making these compositions, and their use, are also described.

Abstract: Cell storage and delivery systems and methods for storing and delivering viable cells to a mammal are disclosed. The cell storage and delivery systems include a biodegradable and/or bioabsorbable fibrous matrix physically associated with viable cells to contain and release the cells at a controlled rate. The biodegradable and/or bioabsorbable matrix can be formed by electrospinning fibers of biodegradable and/or bioabsorbable fiberizable material. The methods include methods for storing viable cells and for delivering viable cells to a mammal using the cell storage and delivery system.

Abstract: A method and apparatus for screening an array of test compounds for bioactivity by contacting an array of test compounds with a detector layer capable of detecting bioactivity, and detecting a detector layer response. The detector layer is comprised of physiologically viable cells. The method and apparatus allow a large number of test compounds to be simultaneously assayed in parallel without the need for complex fluidic devices.

Abstract: Granules that include a protein core are described. The protein core includes a protein matrix which includes a protein mixed together with a starch and optionally sugar such as sucrose. The protein matrix can be layered over a seed particle or the protein core can be homogeneous. The protein can be an enzyme or a therapeutic protein. A barrier layer may surround the protein core, and a coating can be applied to the seed particle, the protein matrix and/or the barrier layer.

Abstract: The present invention includes a process for separating cells from a liquid media, a method of fermentation using such a separation, and an apparatus for conducting such a separation or for facilitating a cellular reaction. In broadest terms, the separation process of the present invention is a process for separating cells from a liquid containing cells. The process comprises the steps of: (a) bringing the liquid containing the cells into contact with one or more microbial polysaccharide and a fibrous material so as to adsorb the cells onto the fibrous material; and (b) separating the liquid from the fibrous material so as to remove the cells from the liquid. The process of the present invention may be used to remove cells from any liquid, but such liquids typically will be aqueous solutions, such as growth media, biological fluids, diagnostic samples, aqueous test samples, etc.

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: Compositions of matter are described which contain restricted cancer cells. When so restricted, the cells produce an unexpectedly high amount of material which suppresses cancer cell proliferation. The phenomenon crosses cancer type and species lines. Processes for making these compositions, and their use, are also described.

Abstract: Disclosed are a composition of chemically defined components which support in vitro and in vivo chondrogenesis of mesenchymal stem cells, a method for in vitro and in vivo chondrogenic induction of such stem cells, and a method of forming human chondrocytes in vitro and in vivo from such stem cells.

Abstract: A polyurethane polymer includes at least one buffer selected to adjust pH to a pH within a desired range. The buffer compound is immobilized within the polymer, and the polymer has a buffer capacity in excess of 3 micromoles of acid or base per gram polymer. A polymer includes at least one enzyme that is selected to catalyze a reaction of a substance. The enzyme is immobilized within the polymer. The polymer also includes at least one buffer selected to adjust the pH in the vicinity of the enzyme to a pH within a desired range. The buffer compound is also immobilized within the polymer. The polymer preferably has enzyme activity and a buffer capacity in excess of 3 micromoles acid or base per gram polymer. In one embodiment, each of the enzyme(s) and the buffer(s) are covalently bonded to the polymer.

Abstract: A method of increasing loading of active enzyme immobilized in a polyurethane polymer including the steps of: synthesizing the polyurethane polymer in a reaction mixture containing water and enzyme; and including a sufficient amount of a surfactant in the reaction mixture to increase enzyme activity at an enzyme loading.

Abstract: The present invention provides a method of biological treatment for purifying an environmental pollutant utilizing microorganisms, wherein the pollutant is efficiently concentrated to a high concentration but exerts substantially no effect on the biological activity of the microorganisms used to biodegrade the pollutant, so that the pollutant are purified (assimilated or degraded) effectively. The present invention also provides a microbial treatment agent useful for the purification treatment of an environmental pollutant. Specifically stated, the method for purification treatment of the present invention can be carried out by making an environmental pollutant and microorganisms to coexist with each other as incorporated in a microorganism-produced polymer. The microbial treatment agent of the present invention comprises microorganisms incorporated in a microorganism-produced polymer.

Abstract: Porous films are provided which include a blend of a film forming polymer and a non-film forming material, the film having a network of pores or channels throughout the film. The porous polymer films are formed between 0° and 80° C. retain porosity at elevated temperatures and are non-friable. A process for preparing porous polymer films and their applications are disclosed.

Abstract: A protein system is described in which a protein is bound within a matrix material that has pores that are sized to achieve excellent properties such as: activity, protein density, and stability. In a preferred embodiment, the pore sizes range from 50 to 400 Å. One protein that has demonstrated surprisingly good results in this system is OPH. This protein is known to degrade organophosphorus compounds such as are found in chemical weapons and pesticides. Novel methods of forming the protein system and methods of making OPH are also described.

Abstract: A process for detecting an analyte which process comprises (a) contacting a sample suspected of containing said analyte with a containment means comprising a barrier which separates signal generating reagents from said sample, in the presence of an element which interacts specifically with said analyte, under conditions whereby interaction between the analyte and the said element results in activation of the signal generating reagents within the containment means on the side of the barrier opposite to the sample, and (b) detecting any signal generated and retained within the containment means from the sample side of the barrier. The process of the invention provides for sensitive detection of very small numbers of analyte materials using measurement techniques which include counting methods such as flow cytometry.

Abstract: The present invention provides fibrin microbeads that are biologically active and comprise extensively cross-linked fibrin(ogen), and a method for preparing the fibrin microbeads. The present invention also provides a composition comprising cells bound to the fibrin microbeads, and methods for culturing and separating cells using the fibrin microbeads of the present invention. Finally, the present invention provides methods for transplanting cells and engineering tissue using the fibrin microbeads of the present invention.

Abstract: A biodegradable, biocompatible porous matrix as a scaffold for tissue engineering cartilage is formed of a copolymer of a polyalkylene glycol and an aromatic polyester such as a polyethylene glycol/polybutylene terephtalate copolymer. A ceramic coating such as a calcium phosphate coating may be provided on the scaffold by soaking the scaffold in a solution containing calcium and phosphate ions. A composite scaffold which may be used as an implant is preferably a two-layer system may be formed having an outer surface of a layer of the porous matrix formed of the copolymer, and an outer surface of a layer of a ceramic material. The composite scaffold may be prepared by casting the copolymer on top of the ceramic material in a mould. Cells are preferably seeded on the scaffold prior to implanting, and the scaffold may contain bioactive agents that are released on degradation of the scaffold in vivo.

Abstract: Slowly polymerizing polysaccharide hydrogels have been demonstrated to be useful as a means of delivering large numbers of isolated cells via injection. The gels promote engraftment and provide three dimensional templates for new cell growth. The resulting tissue is similar in composition and histology to naturally occurring tissue. This method can be used for a variety of reconstructive procedures, including custom molding of cell implants to reconstruct three dimensional tissue defects, as well as implantation of tissues generally.

Abstract: An enzymatic catalyst in the granular form which is non-dispersible in water comprising an inner core provided with a coating which coating comprises an enzymatic material characterised in that the coating has a thickness of from 30 to 400 microns and the ratio of the size of the core to the thickness of the coating is from 2 to 20. A method for the preparation of the catalyst and the use of the catalyst in the conversion of 2-hydroxy-4-methylthio butyronitrile to ammonium 2-hydroxy-4-methylthio butanoate is also claimed.

Abstract: The present invention provides a drug delivery device comprising encapsulated cells that contain multiple copies of an expression cassette. The drug delivery device of the present invention is useful to supply an animal, including a human, with a therapeutically desirable molecule, including physiological activities lacking in disease conditions or antagonist against conditions in an animal, especially a human.

Abstract: The invention provides a sewage purification assist toilet paper that carries pineapple enzyme and at least one powdery additive selected from slag, porous ore, and activated carbon. The pineapple enzyme and additive decompose salinity and chlorides contained in toilet sewage which otherwise would inhibit the growth of microorganisms used in the biological treatment of sewage taking place not only in individual septic tanks but also in public sewage systems.

Abstract: This invention provides a process for the production of molded particles for microbial cell-immobilization which is characterized in that: an aqueous liquid composition which comprises: (a) an ethylenically unsaturated group-containing urethane resin which is obtained by making a tertiary amino group-containing diol react with a polyisocyanate compound, and by further introducing an ethylenically unsaturated group; (b) a polymerization initiator; and (c) water-soluble macromolecular polysaccharides; is gelatinized, in an aqueous medium which contains metal ion, to be particulate gel which is then subjected to photopolymerization and/or thermopolymerization by which to cause the crosslinking reaction of ethylenically unsaturated group in said particulate gel.

Abstract: A cell delivery device is prepared comprising a controllable degradable gel phase, meshed within a polymer substrate for use in tissue-engineering. The gel phase comprises a degradable, natural or synthetic polymer, and includes a suspension of living cells. The polymer substrate comprises a biocompatible, degradable polymer, and may be synthetic or natural. Degradation of the gel phase may be controlled by enzyme activity or adjustment of gel phase physical properties. In one embodiment, the gel phase contains an enzyme and/or enzyme inhibitor to control degradation of the gel phase. The device is useful in tissue replacement and repair, and more particularly, in the repair of cartilage tissue.

Abstract: A biodegradable, biocompatible porous matrix as a scaffold for tissue engineering cartilage is formed of a copolymer of a polyalkylene glycol and an aromatic polyester such as a polyethylene glycol/polybutylene terephtalate copolymer. A ceramic coating such as a calcium phosphate coating may be provided on the scaffold by soaking the scaffold in a solution containing calcium and phosphate ions. A composite scaffold which is preferably a two-layer system may be formed having an outer surface of a layer of the porous matrix formed of the copolymer, and an outer surface of a layer of a ceramic material. The composite scaffold may be prepared by casting the copolymer on top of the ceramic material in a mould. Cells are preferably seeded on the scaffold prior to implanting, and the scaffold may contain bioactive agents that are released on degradation of the scaffold in vivo.

Abstract: Immobilized garlic alliinase wherein the alliinase is chemically, physically or biologically immobilized, is useful in a method for continuous production of allicin. The method comprises adding a solution of alliin as substrate to a column containing the immobilized garlic alliinase and collecting pure allicin in the effluent. The pure allicin is intended for use as food additive or for the preparation of pharmaceutical compositions for the treatment of viral, bacterial, fungal and parasitic infections, high levels of cholesterol and blood lipids, high blood pressure and thrombosis.

Abstract: A fibrous polymer implant loaded with one or more bioactive agents is prepared using a wet spinning technique. Preferably, an aqueous solution of bioactive agent is added to a solution of amphiphilic block copolymer containing hydrophilic blocks such as polyalkylene glycol and hydrophobic blocks such as an aromatic ester dissolved in a first solvent immiscible with water to form an emulsion. The emulsion is injected through a nozzle into a second solvent miscible with the first solvent in which the copolymer is essentially insoluble to form a solid copolymer fiber loaded with the bioactive agent. The fiber is shaped into an implant. Water content of the aqueous solution of bioactive agent affects rate of release of the bioactive agent in vivo. Bioactive agents include peptides, oligopeptides, polypeptides and proteins. The implant may be used as a carrier for controlled drug release or as a scaffold for tissue engineering.

Abstract: Biomacromolecules encapsulated or immobilized within mesoporous or nanoporous materials via surfactant-templated sol-gel reactions and methods for producing these encapsulated 5 or immobilized biomacromolecules are provided.

Abstract: A temperature-stable droplet is provided containing a temperature-stable hydrocolloid membrane. The hydrocolloid membrane encapsulates a liquid that contains at least one enzyme, a cell, a biological agent, a pharmaceutical agent, an immunological agent, or mixtures thereof, and at least one of a locust bean gum, a natural thickening agent, a guar, polyvinylpyrrolidone, Konjac mannan, methylcellulose, hydroxymethylcellulose, calcium gluconate, glucomannan, or mixtures thereof. Preferably, the hydrocolloid membrane comprises at least one of methoxy pectin, Konjac mannan, sodium alginate, or mixtures thereof, and at least one of a locust bean gum, methylcellulose, hydroxymethylcellulose, glucomannan, or mixtures thereof. The hydrocolloid membrane encapsulating the liquid is a thickness capable of holding the liquid without bursting through a temperature range of about −20° C. to about 90° C.

Abstract: A method for preserving immobilized or unimmobilized microbial cells having nitrilase activity and for stabilizing the nitrilase activity of unimmobilized or immobilized microbial cells has been developed. Aqueous suspensions containing at least 100 mM bicarbonate, carbonate, or carbamate salts limit microbial contamination of the stored enzyme catalyst, as well as stabilize the desired nitrilase activity of the unimmobilized or immobilized cells. Microorganisms which are characterized by an nitrilase activity and are stabilized and preserved by this method include Acidovorax facilis 72-PF-15 (ATCC 55747), Acidovorax facilis 72-PF-17 (ATCC 55745), Acidovorax facilis 72W (ATCC 55746), and transformed microbial cells having nitrilase activity, the host cells transformed with Acidovorax facilis 72W nitrilase activity. Especially preferred is an embodiment using ammonium carbamate as the inorganic salt.

Abstract: Methods for detecting allelic variants of the myostatin (growth and differentiation factor-8) gene are provided. Specifically provided are methods of identifying subjects having or having a predisposition for increased muscle mass as compared to subjects having wild-type myostatin. Increased muscle mass is particularly desirable for identification of animals used to produce food products, including bovine, porcine, ovine, avian and piscine species.

Abstract: A method and associated apparatus for use in a microbiological assay procedure wherein a plurality of diffusion disks are placed on a nutrient medium on a plate, each of the disks carrying a respective antibiotic agent and each having an identification code identifying the respective antibiotic agent, the disks being positioned on the plate in a preselected relative arrangement. The method comprises optically scanning the disks on the plate, consequently generating a digitally encoded image including digitized representations of the identification codes, and electronically processing the digitally encoded image to determine an angle of rotation of the plate relative to a pre-established reference frame or coordinate system, wherein the electronic processing of the digitally encoded image includes detecting a region of the plate occupied by a unique subset of the disks.