Abstract: The present disclosure provides a method for culturing cells in exogenous lactic acid. Certain aspects of the present disclosure include the production of recombinant proteins, such as antibodies and fragments thereof. Certain aspects of the present disclosure also relate to methods of controlling lactic acid production, pH stability and osmolality in cell culture.

Abstract: Novel promoters which are derived from P. pastoris pastoris which are inducible or repressible under specific growth conditions are provided. These promoters are useful for regulating the expression of a desired structural gene, e.g., a mammalian polypeptide. Particularly preferred is the use of these novel promoters to regulate gene expression in polyploidal yeast such as diploidal P. pastoris produced by mating or spheroplast fusion.

Abstract: A preferred embodiment of the present invention provides a method and system for building cost-efficient biocompatible hydrogel constructs using stereolithography. Hydrogel constructs may be used in, for example, multi-lumen nerve regeneration conduits and other tissue engineering scaffolds with embedded channel architecture that facilitate tissue regeneration through possible incorporation of precisely located bioactive agents, cells, and other desired inert and/or active chemical agents and devices. Another preferred embodiment of the present invention provides a method of fabricating a hydrogel construct comprising: solidifying a first solution into a first construct layer with a first energy dosage using stereolithography, the first solution comprising: a first polymer; and a first photoinitiator, wherein the first polymer and first photoinitiator are of a first concentration.

Abstract: The present invention provides a novel method to isolate and expand pure progenitor/stem cells from a primary tissue explant, which produces a population enriched in multipotent functional progenitor/stem cells free of contaminating fibroblasts and other cell types. Cardiac progenitor/stem cells isolated by this method maintain their self-renewal and clonogenic character in vitro and differentiate into normal cells in myocardium, including cardiomyocytes, endothelial cells, and smooth muscle cells, after transplantation into ischemic hearts. The present invention also includes substantially pure populations of multipotent progenitor/stem cells, e.g., cardiac progenitor/stem cells, and their use to treat and prevent diseases and injuries, including those resulting from myocardial infarction.

Abstract: The present invention provides a novel method to isolate and expand pure progenitor/stem cells from a primary tissue explant, which produces a population enriched in multipotent functional progenitor/stem cells free of contaminating fibroblasts and other cell types. Cardiac progenitor/stem cells isolated by this method maintain their self-renewal and clonogenic character in vitro and differentiate into normal cells in myocardium, including cardiomyocytes, endothelial cells, and smooth muscle cells, after transplantation into ischemic hearts. The present invention also includes substantially pure populations of multipotent progenitor/stem cells, e.g., cardiac progenitor/stem cells, and their use to treat and prevent diseases and injuries, including those resulting from myocardial infarction.

Abstract: The present disclosure provides a method for culturing cells in exogenous lactic acid. Certain aspects of the present disclosure include the production of recombinant proteins, such as antibodies and fragments thereof. Certain aspects of the present disclosure also relate to methods of controlling lactic acid production, pH stability and osmolality in cell culture.

Abstract: Methods are provided for the synthesis and secretion of recombinant proteins preferably large mammalian proteins or hetero-multimeric proteins at high levels and for prolonged time in polyploid, preferably diploid yeast. These methods use various mating competent yeast, including Pichia. In a preferred embodiment, a first expression vector is transformed into a first haploid cell; and a second expression vector is transformed into a second haploid cell. The transformed haploid cells, each individually synthesizing a non-identical polypeptide, are identified and then genetically crossed or fused. The resulting diploid strains are utilized to produce and secrete fully assembled and biologically functional hetero-multimeric protein.

Abstract: A preferred embodiment of the present invention provides a method and system for building cost-efficient biocompatible hydrogel constructs using stereolithography. Hydrogel constructs may be used in, for example, multi-lumen nerve regeneration conduits and other tissue engineering scaffolds with embedded channel architecture that facilitate tissue regeneration through possible incorporation of precisely located bioactive agents, cells, and other desired inert and/or active chemical agents and devices. Another preferred embodiment of the present invention provides a method of fabricating a hydrogel construct comprising: solidifying a first solution into a first construct layer with a first energy dosage using stereolithography, the first solution comprising: a first polymer; and a first photoinitiator, wherein the first polymer and first photoinitiator are of a first concentration.

Abstract: Oscillating angularly rotating a container containing a material may cause the material to be separate. Denser or heavier material may unexpectedly tend to collected relatively close to the axis of rotation, while less dense or light material may tend to collect relatively away from the axis of rotation. Oscillation along an arcuate path provides high lysing efficiency. Alternatively, a micromotor may drive an impeller removably received in a container. Lysing may be implemented in batch mode, flow-through stop or semi-batch mode, or flow-through continuous mode. Lysing particulate material may exceed material to be lysed or lysed material and/or air may be essentially eliminated from a chamber to increase lysing efficiency.

Abstract: The present invention provides a novel method to isolate and expand pure progenitor/stem cells from a primary tissue explant, which produces a population enriched in multipotent functional progenitor/stem cells free of contaminating fibroblasts and other cell types. Cardiac progenitor/stem cells isolated by this method maintain their self-renewal and clonogenic character in vitro and differentiate into normal cells in myocardium, including cardiomyocytes, endothelial cells, and smooth muscle cells, after transplantation into ischemic hearts. The present invention also includes substantially pure populations of multipotent progenitor/stem cells, e.g., cardiac progenitor/stem cells, and their use to treat and prevent diseases and injuries, including those resulting from myocardial infarction.

Abstract: Novel promoters which are derived from P. pastoris pastoris which are inducible or repressible under specific growth conditions are provided. These promoters are useful for regulating the expression of a desired structural gene, e.g., a mammalian polypeptide. Particularly preferred is the use of these novel promoters to regulate gene expression in polyploidal yeast such as diploidal P. pastoris produced by mating or spheroplast fusion.

Abstract: A symbology or mapping between multi-dimensional topographical parameters and characters allows multi-dimensional topographical problems to be turned into one-dimensional sequence problems, allowing fast, computationally efficient and robust sequence analysis methodologies to be employed. Substitution matrices allow the scoring of matches or hits. Such techniques may be applied to proteins, polymers or other multi-dimensional structures.

Abstract: An active matrix microfluidic platform employs thin film transistor active (“TFT”) matrix liquid crystal display technology to manipulate small samples of fluid for chemical, biochemical, or biological assays without moving parts, for example, using a two-dimensional matrix array of drive electrodes. The active matrix microfluidic platform may employ existing active matrix addressing schemes and/or commercial “off-the-shelf” animation software to program assay protocols. A feedback subsystem may determine an actual location of a fluid in the microfluidic structure, and provides location information to for display, for example, on an active matrix display, and/or to control movement of one or more fluid bodies in the microfluidic structure.

Abstract: Methods are provided for the synthesis and secretion of recombinant proteins preferably large mammalian proteins or hetero-multimeric proteins at high levels and for prolonged time in polyploid, preferably diploid yeast. These methods use various mating competent yeast, including Pichia. In a preferred embodiment, a first expression vector is transformed into a first haploid cell; and a second expression vector is transformed into a second haploid cell. The transformed haploid cells, each individually synthesizing a non-identical polypeptide, are identified and then genetically crossed or fused. The resulting diploid strains are utilized to produce and secrete fully assembled and biologically functional hetero-multimeric protein.

Abstract: The present invention provides methods and kits for amplifying target nucleic acids (including whole genomes) using nicking agents. In certain aspects, the amplification does not require the use of any external oligonucleotide primers that are capable of annealing to a portion of the target nucleic acid. This invention is useful in many areas such as genetic disease diagnoses, forensic analyses and palcoarcheological studies.

Abstract: Compositions and methods for identifying agents that inhibit protease activity are provided. In particular, polynucleotides, recombinant expression vectors, and host cells are provided that may be used in a bacterial cell-based assay for identifying agents that are inhibitors of protease activity, such as inhibitors of HIV protease activity. The bacterial cells express a precursor of a protease and encode a reporter polypeptide that contains a protease recognition sequence, which can be cleaved by the mature, catalytically active protease such that the reporter activity of the reporter polypeptide is decreased or eliminated.

Abstract: The present invention provides methods and compositions for nucleic acid methylation analysis using nicking agents.

Abstract: An microfluidic platform employs a two-dimensional matrix array of drive electrodes and at least one ground line on a bottom substrate, eliminating the need for a top plate or cover, to allow easy access to the active surface of the microfluidic platform. The open microfluidic platform may, for example, allow the depositing of samples via an array of pipettes or other automated deliver systems, and/or the use of standard video equipment to focus on the active surface to track positions of fluid bodies. A user may move fluid bodies and perform operations in real time and/or create animation files for later execution using a pointing device and a display device such as a monitor.

Abstract: The invention provides a method for identifying a nucleotide at a defined position in a target nucleic acid using restriction endonucleases and fluorescence polarization. The invention further provides compounds, compositions, and kits related to the method.

Abstract: The present invention provides methods and compositions for gene expression analyses using nicking agents.