Abstract: The present invention relates to a method of determining the sequence and/or occurrence frequency of a number of variable gene inserts from a gene library, which inserts exhibit a desired specific characteristic, wherein each variable gene insert is flanked (5?) and (3?) by known sequences, the method comprising; selecting the number of inserts by their ability to exhibit the desired specific characteristic, conducting polymerase chain reaction to amplify the selected number of variable gene inserts to produce components of a mixed PCR product, ligating the components of the mixed PCR product to produce a concatenated sequence and sequencing or determining the occurrence of the gene inserts in the concatenated sequence.

Abstract: Translationally competent ribosomes are bound to a solid substrate through a specific attachment site. A spatial array of such immobilized ribosomes may be produced on a planar substrate, microbeads, on fiber optics. One or more components of the ribosome complex are preferably labeled, e.g. with a fluorescent dye. Ribosomal RNAs, including mRNA and tRNA; and/or ribosomal proteins may be labeled at specific positions, and arrays of immobilized ribosomes may comprise a panel of different labels and positions of labels. Fluorescence detection can then be used to monitor conformational dynamics and translation rates.

Abstract: This invention relates to a process by which an extract comprising integral thylakoids is obtained. The resulting extract is a potent dynamic antioxidant useful as a ROS (reactive oxygen species) scavenger. This extract is intended to be used for the treatment or prevention of diseases involving the generation of ROS, such as inflammatory diseases or cancer. This extract also finds a use as a solar screen because of its capacity to capture UV radiations and to dissipate the solar energy into heat.

Abstract: The present invention relates to therapeutic agents useful for the treatment of Severe Acute Respiratory Syndrome (SARS) in humans. In particular, the present invention relates to RNA interference (RNAi) molecules useful for inhibiting the infection and replication of hSARS virus. Preferably, the RNAi molecules target the replicase region of the hSARS virus, or combinations of different sites of hSARS virus genes. The present invention further encompasses methods of using the RNAi molecules for preventing and/or treating SARS. Vaccines and kits comprising therapeutically effective amounts of the RNAi molecules are also encompassed.

Abstract: Provided are a method of producing a culturable cell with no nucleus, showing mitochondrial activity, comprising: performing cell fusion between a nucleus-less cell having mitochondrial DNA and a mitochondrial DNA-less cultured cell derived from a cancer cell; culturing resulting cybrid cells; and recovering floating cells from obtained cultured cells, and a cell obtained by the method.

Abstract: This invention features in vitro methods of determining the rate and extent to which test compounds, such as drug candidates, enter, move within, and traverse lipid-associated barriers. This invention also features method that measure the accumulation, that is the partition coefficient, of test compounds in lipid structures relative to, e.g., an aqueous phase, as well as the rate of accumulation of test compounds in the lipid structures. In addition, the invention features lipid vesicles that contain fluorophores localized within the aqueous interior that can be employed in the methods described herein.

Abstract: A method for removing a gas from a site comprising placing cells having gas vesicles under conditions that induce the cells to float to a surface of an aqueous medium, harvesting the cells from the surface of the medium, lysing the cells, separating the gas vesicles from the lysed cells, crosslinking the gas vesicles with a crosslinking agent, loading a gas with a lowered partial pressure for the compound to be removed into the gas vesicles, and placing the gas vesicles such that the gas compound is removed from the site. Harvesting gas-vesicle-containing cells is achieved by placing the cells under conditions that induce the cells to rise to the surface of an aqueous medium—such as darkness, exponential growth stage, flocculation, or dissolved gas flotation—then collecting the cells from the surface of the medium. Gas vesicles are isolated by lysing the cells and separating the gas vesicles from the lysate. Once the gas vesicles are isolated, they can be modified, such as by crosslinking with glutaraldehyde.

Abstract: Safe and effective live vaccines against Flavobacterium columnare of fish were created through the induction of rifampicin resistance in a native Flavobacterium columnare isolate; these including rifampicin-resistant mutants NRRL B-30303 and B-30304. Single immersion exposure of fish stimulated acquired immunity against virulent F. columnare infection.

Abstract: Disclosed is a method and apparatus for the isolation of DNA or cell nuclei or a mixture thereof from cell samples in a CD device. The method includes treating a suspension of whole cells with a lysis reagent so as to lyse the cytoplasmic membrane and at least some of the nuclear membranes, and introducing the lysate into micro-channels of a microfabricated apparatus in which each of the micro-channels is provided with a barrier disposed in the channel to impede the passage or flow of DNA and cell nuclei while allowing the passage of liquid through the micro-channel so that a mesh comprising DNA is formed in the channel.

Abstract: This invention relates to a method of preparing membrane vesicles from a biological sample, characterised in that it comprises at least one anion exchange and/or gel permeation chromatography treatment of the sample. The invention is used to prepare vesicles of varied origins and types, particularly from antigen presenting cells or tumoral cells. The invention also relates to the vesicles obtained in this way and their uses.

Abstract: There has been invented an apparatus comprising a separation barrier for excluding denser cell materials from less dense cell materials after centrifuging of the cells so that selected materials can be withdrawn from the less dense cell materials without inclusion of the denser cell materials or clogging of sampling equipment with denser cell materials. Cells from which selected material is to be withdrawn are centrifuged, either as cells or cells in media. Once the denser cell materials are isolated in a layer by centrifugal force, an invention screen or seive is submerged in the less dense cell material to a level above the layer of denser cell materials to isolate the denser cell materials from the less dense cell materials, preventing mixing of the denser cell materials back into the less dense cell materials when the cells or the cells in media are no longer being centrifuged and to prevent clogging of sampling equipment with denser cell materials.

Abstract: Disclosed is a method and apparatus for the isolation of DNA or cell nuclei or a mixture thereof from cell samples in a CD device. The method includes treating a suspension of whole cells with a lysis reagent so as to lyse the cytoplasmic membrane and at least some of the nuclear membranes, and introducing the lysate into micro-channels of a microfabricated apparatus in which each of the micro-channels is provided with a barrier disposed in the channel to impede the passage or flow of DNA and cell nuclei each while allowing the passage of liquid through the micro-channel so that a mesh comprising DNA is formed in the channel.

Abstract: Disclosed is a composition that includes a material that is susceptible to degradation and a preserving agent in an amount effective to preserve the material comprising one or more reduced malto-oligosaccharide species. The preserving agent can include a single reduced malto-oligosaccharide species or a plurality of such species. Further disclosed is a method of preserving a material. The method generally includes contacting the material with a preserving agent containing a preserving effective amount of one or more reduced malto-oligosaccharide species. Solutions, powders, glasses, gels, and the like containing the chemically reactive material(s) and a preserving effective amount of one or more reduced malto-oligosaccharide species may be prepared.

Abstract: The present invention relates to the field of glycosylation engineering of proteins. More particularly, the present invention relates to nucleic acid molecules, including fusion constructs, having catalytic activity and the use of same in glycosylation engineering of host cells to generate polypeptides with improved therapeutic properties, including antibodies with increased Fc receptor binding and increased effector function.

Abstract: Fluid planar lipid layer-based membrane-anchored ligand systems with defined ligand valency and methods of use thereof are provided.

Abstract: A method for loading a biological sample comprising loading a biological sample with a solute and dimethylsulfoxide (DMSO) by fluid phase endocytosis to produce an internally loaded biological sample. A dehydrated composition is provided that includes dried biological samples containing dimethylsulfoxide and a solute. A method for increasing the survival of biological samples comprising providing biological samples, loading the biological samples with a carbohydrate and dimethylsulfoxide to produce loaded biological samples, and drying (e.g., air drying or vacuum drying) the loaded biological samples while maintaining a residual water content in the biological samples of at least about 0.01 gram water per gram of dry weight of biological samples to increase survival of the biological samples upon rehydrating.

Abstract: Novel chimeric constructions and methods for their use are provided for expression of exogenous genes in a plant chloroplast. Particularly, expression is achieved by the use of a chloroplast or bacterial 5′ untranslated region in the expression cassette. The expression cassette may be integrated into the chloroplast genome by the use of chloroplast DNA flanking sequences, or may replicate autonomously if provided with a chloroplast origin of replication. Plants and cells containing the transformed chloroplasts are also provided. The constructs may be used with both monocotyledenous and dicotyledenous chloroplasts.

Abstract: A tissue graft composition comprising liver basement membrane is described. The graft composition can be implanted to replace or induce the repair of damaged or diseased tissues.

Abstract: A method for loading a biological sample comprising loading a biological sample with a solute by fluid phase endocytosis to produce an internally loaded biological sample. Within the biological sample a first matter (e.g., a vesicle) having the solute fuses with a second matter (e.g., a lysosome) to produce a fused matter containing the solute. Loading of the biological sample includes transferring the solute from the fused matter into cytoplasm within the biological sample.

Abstract: Methods and apparatus for recovering dental pulp from dentition of a donor are disclosed, wherein the pulp from within extracted teeth utilizing such methods and apparatus is harvested, while preserving a sterile environment and avoiding trauma and infection, and stem cells, dendritic cells, and other cells isolating from the pulp, and the various cells propagated and expanded for subsequent use in repair or regeneration of tissues of the body, for therapeutic treatments, and other medical purposes.

Abstract: A synthetic, nuclear-encoded ND4 gene was linked to a mitochondrial targeting sequence and a FLAG epitope tag. This fusion construct was inserted into a rAAV vector. The ND4 fusion protein was expressed and imported into the mitochondria of cells harboring a mitochondrial DNA mutation (G11778A), where it restored cellular respiration.

Abstract: The invention relates to a nucleic acid that encodes a plant transporter of the inner mitochondrial membrane, in particular a member of the mitochondrial carrier family (MCF), and uses of this nucleic acid. The invention furthermore relates to a fragment of the nucleic acid, a construct containing the nucleic acid or a fragment thereof, and a host cell that contains the nucleic acid, the fragment, or the construct. The present invention furthermore relates to a method for producing a transgenic plant by using the nucleic acid, a fragment thereof, or a construct containing the nucleic acid or a fragment thereof, as well as a method for modulating the transport properties of the inner mitochondrial membrane of a plant, a plant part, a plant cell and/or seeds.

Abstract: Provided are a method of producing a culturable cell with no nucleus, showing mitochondrial activity, comprising: performing cell fusion between a nucleus-less cell having mitochondrial DNA and a mitochondrial DNA-less cultured cell derived from a cancer cell; culturing resulting cybrid cells; and recovering floating cells from obtained cultured cells, and a cell obtained by the method.

Abstract: The invention of the present application provides an isolated and purified human mitochondrial protein comprising the amino acid sequence of SEQ ID NO: 2 or SEQ ID NO: 4, which is a novel human protein promoting aggregation and fusion of mitochondria. The present invention also provides a polynucleotide encoding such a mitochondrial protein, antibody against such a mitochondrial protein, and a proteoliposome composed of such a mitochondrial protein and lipid. Mitochondrial proteins are useful for clarifying causes of mitochondrial diseases as well as for developing preventive and therapeutic methods thereof. Furthermore, antibodies and probes derived from genes encoding such proteins are potentially useful materials for diagnosis of condition of mitochondria in particular diseases. Furthermore, proteoliposomes provide measures for specific transfer of foreign genes or drugs targeted toward mitochondria.

Abstract: A cell in which the activity of a protein relating to transport of an intracellular sugar nucleotide, GDP-fucose, to the Golgi body is more decreased or deleted than its parent cell; a process for producing an antibody composition using the cell; a transgenic non-human animal or plant or the progenies thereof, in which genome is modified so as to have a decreased or deleted activity of a protein relating to transport of an intracellular sugar nucleotide, GDP-fucose, to the Golgi body; a process for producing an antibody composition from the animal or plant; and a medicament comprising the antibody composition.

Abstract: Mitochondrial targets for drug screening assays and for therapeutic intervention in the treatment of diseases associated with altered mitochondrial function are provided. Complete amino acid sequences [SEQ ID NOS:1-3025] of polypeptides that comprise the human heart mitochondrial proteome are provided, using fractionated proteins derived from highly purified mitochondrial preparations, to identify previously unrecognized mitochondrial molecular components.

Abstract: The present invention provides methods for introducing functional peptides into organelles. Additionally, the present invention provides a method for correcting a phenotypic deficiency in a mammal that results from a mutation in the mammal's mitochondrial DNA (mtDNA). The present invention further provides a method for treating a mitochondrial disorder in a subject in need of treatment therefor. Also provided is an expression vector that is useful for introducing a functional peptide encoded by an mtDNA sequence into a mitochondrion. The present invention also provides eukaryotic cells transformed by expression vectors that are useful for introducing functional peptides into organelles. Finally, the present invention provides a pharmaceutical composition comprising a non-nuclear nucleic acid sequence encoding a peptide for introduction into an organelle, a nucleic acid sequence encoding an organelle-targeting signal, and a pharmaceutically-acceptable carrier.

Abstract: A biocompatible biological component is provided comprising a membrane-mimetic surface film covering a substrate. Suitable substrates include hydrated substrates, e.g. hydrogels which may contain drugs for delivery to a patient through the membrane-mimetic film, or may be made up of cells, such as islet cells, for transplantation. The surface may present exposed bioactive molecules or moieties for binding to target molecules in vivo, for modulating host response when implanted into a patient (e.g. the surface may be antithrombogenic or antiinflammatory) and the surface may have pores of selected sizes to facilitate transport of substances therethrough. An optional hydrophilic cushion or spacer between the substrate and the membrane-mimetic surface allows transmembrane proteins to extend from the surface through the hydrophilic cushion, mimicking the structure of naturally-occurring cells.

Abstract: A soluble derivative of a soluble polypeptide, which comprises two or more heterologous membrane binding elements with low membrane affinity covalently associated with the polypeptide, the elements being soluble in aqueous solution, and the elements being capable of interacting, independently and with thermodynamic additivity, with components of cellular or artificial membranes exposed to extracellular fluids, characterized in that the membrane binding elements target lipid raft components of the membrane and bind to the lipid rafts to localize the polypeptide at the lipid rafts.

Abstract: The object of the invention is a novel sensitization process for antigen-presenting cells, novel means for implementing the process and novel membrane vesicles possessing immunogenic potency. The invention relates in particular to texosomes (vesicles derived from tumor cells) and dexosomes (vesicles derived from dendritic cells loaded or not with antigens), and their use for the vectorization and presentation of antigens in vitro or in vivo as well as in methods or compositions for the treatment of cancers and infectious, parasitic or autoimmune diseases in particular.

Abstract: The present invention is directed to a tubular nanostructure for providing a stable nanometer-sized pore across a lipid bilayer membrane having a hydrophobic core region between two hydrophilic surface regions comprising a tubular body having a hydrophobic region flanked by hydrophilic regions, a method for inserting a tubular nanostructure into a lipid bilayer membrane, and a method for providing a stable pore in a lipid bilayer membrane.

Abstract: Methods are provided for generating mobile, membrane-associated biomolecules in a supported membrane, by the process of sequestering and collecting mobile and immobile populations of biomolecules. Populations of mobile biomolecules, but not those that are immobile, can be moved to a region through a variety of mechanisms, including passive diffusion or induced drift, generating a pure, mobile population of biomolecules. In some embodiments of the invention, complex substrates are utilized, e.g. substrates containing switchable barriers to lipid diffusion, including fluidic channels; removal of regions of lipid bilayer by blotting, and the like.

Abstract: The invention concerns membrane fractions of cells containing a recombinant MGDG synthase and enriched with 1,2-sn-diacylglycerol, their preparation method, their use for screening molecules inducing MGDG synthase activity and a method for screening molecules inducing MGDG synthase activity using said membrane fractions.

Abstract: Targeted therapeutics that localize to a specific subcellular compartment such as the lysosome are provided. The targeted therapeutics include a therapeutic agent and a targeting moiety that binds a receptor on an exterior surface of the cell, permitting proper subcellular localization of the targeted therapeutic upon internalization of the receptor. Nucleic acids, cells, and methods relating to the practice of the invention are also provided.

Abstract: The invention provides compositions and methods for the production of achromosomal and anucleate cells useful for applications such as diagnostic and therapeutic uses, as well as research tools and agents for drug discovery.

Abstract: The invention provides chimeric cannulae polypeptides and methods for making and using them. In one aspect, the invention provides compositions and methods for the identification, separation or synthesis of proteins or ligands. In one aspect, the invention provides compositions and methods for making and using nanotubules. In one aspect, the invention provides compositions and methods for the selection and purification of chiral compositions from racemic mixtures.

Abstract: The invention provides a method for selecting a population of non-cellular physical entities. The method involves applying energy to one or more non-cellular physical entities having selected parameter signatures, each physical entity located at specific coordinates in a domain and contained within a population of physical entities, thereby altering a property of the one or more physical entities, wherein the alteration renders the one or more physical entities separable from other members of the population of physical entities. The invention also provides a methods for preparing a population of uniquely tagged non-cellular physical entities, and methods for preparing a population of uniquely tagged probes. The invention further provides a method for simultaneously detecting a plurality of analytes using the uniquely tagged probes.

Abstract: The present invention relates to improved diagnostic methods for early detection of a risk for developing type 2 diabetes mellitus in humans, and screening assays for therapeutic agents useful in the treatment of type 2 diabetes mellitus, by comparing the levels of one or more indicators of altered mitochondrial function. Indicators of altered mitochondrial function include enzymes such as mitochondrial enzymes and ATP biosynthesis factors. Other indicators of altered mitochondrial function include mitochondrial mass, mitochondrial number and mitochondrial DNA content, cellular responses to elevated intracellular calcium and to apoptogens, and free radical production. Methods of treating, and of stratifying, human patients as such methods relate to disclosed indicators of altered mitchondrial function are also provided.

Abstract: The present invention provides methods for producing high resolution crystals of ribosomes and ribosomal subunits as well as the crystals produced by such methods. The x-ray diffraction patterns of the crystals provided by the present invention are of sufficiently high resolution for determining the three-dimensional structure of ribosomes and ribosomal subunits, for identifying ligand binding sites on ribosomes and ribosomal subunits, and for molecular modeling of ligands which interact with ribosomes and ribosomal subunits. The present invention provides methods for identifying ribosome-related ligands and methods for designing ligands with specific ribosome-binding properties. Thus, the methods of the present invention may be used to produce ligands which are designed to kill or inhibit any specific target organism(s).

Abstract: The invention relates generally to methods of modulating cell migration. Included in the methods of the invention are methods of identifying the state of a pseudopodium in cell migration and methods of inducing extension or retraction of a pseudopodia from a cell. The invention also relates to methods of screening for and identifying an agent effective in inducing extension or retraction of a pseudopodium and therefore affecting cell migration. Agents that can modulate cell migration are useful in treatment of conditions in which cell migration plays a role. Such conditions can include wound healing, angiogenesis, and metastasis of a disease from one location to another. Additionally, the invention provides methods of biochemically separating the pseudopodium of a cell from the remainder of the cell body and methods of determining the proteins present in the pseudopodium and cell body. The invention also includes a pseudopodium isolated by the methods of the invention.

Abstract: Natural and modified neurotoxins and isolated neurotoxin compositions are described. The neurotoxins may include one or more structural modifications, wherein the structural modification(s) alters the biological persistence, such as the biological half-life and/or a biological activity of the modified neurotoxin relative to an identical neurotoxin without the structural modification(s). In one embodiment, methods of making the modified neurotoxin include using recombinant techniques. In another embodiment, methods of using the modified neurotoxin to treat conditions include treating various disorders, neuromuscular aliments and pain.

Abstract: Translationally competent ribosomes are bound to a solid substrate through a specific attachment site. A spatial array of such immobilized ribosomes may be produced on a planar substrate, microbeads, on fiber optics; and the like. One or more components of the ribosome complex are preferably labeled, e.g. with a fluorescent dye. Ribosomal RNAs, including mRNA and tRNA; and/or ribosomal proteins may be labeled at specific positions, and arrays of immobilized ribosomes may comprise a panel of different labels and positions of labels. Fluorescence detection can then be used to monitor conformational dynamics and translation rates.

Abstract: A composition comprising a novel Ca2+-activated, [ATP]i-sensitive nonspecific cation (NCCa-ATP) channel is described. The channel is found in mammalian neural cells and exhibits a different sensitivity to block by various adenine nucleotides, and is activated by submicromolar [Ca]i. The NCCa-ATP channel is activated under conditions of ATP depletion, which causes severe cell depolarization, followed by cell swelling. The NCCa-ATP channel is regulated by a sulfonylurea receptor and is inhibited by sulfonylurea compounds glibenclamide and tolbutamide. Methods employing compositions comprising the NCCa-ATP channel to screen for compounds that block the channel and the use of such antagonists as therapeutics in preventing brain swelling and damage are described. In addition, methods employing compositions comprising the Kir2.3 channel to screen for compounds that open the channel and the use of such antagonists as therapeutics in preventing brain swelling and damage are described.

Abstract: A method of generating a plant tolerant of environmental stress conditions, the method comprising the step of expressing within the plant exogenous polynucleotides encoding enzymes capable of catalyzing proline production, wherein the exogenous polynucleotides are expressed in a manner so as to allow accumulation of the enzymes within a subcellular organelle of the plant. The invention further describes novel polynucleotides and nucleic acid constructs useful in implementing the methods and a transgenic plant resultant therefrom.

Abstract: The present invention is directed toward antibodies specific for particular intracellular membrane components and the method of using these antibodies. In one aspect of the invention, injecting isolated membrane components into mice may generate specific antibodies to particular membrane components. The injection may include a membrane component by itself or in combination with another membrane component using an immunosuppression method. In one aspect of the invention, these antibodies are used as a novel method and system for drug delivery targeted to particular intracellular membrane components. Another aspect of the invention involves the use of these antibodies for identifying particular intracellular membrane components important for cell functions.

Abstract: The invention provides a novel vehicle for vaccination, in particular peptide vaccination. The new vehicle has been termed an exosome. Exosomes are vesicles derived from MHC class II enriched compartments in antigen presenting cells. The exosomes possess MHC II and/or MHC I molecules at their surface and possibly peptides derived from processed antigens in said MHC's. Thus the exosome is a perfect vaccination vehicle in that it presents the peptide in a natural setting. The peptides present in the exosome in the MHC molecule may be processed by the antigen presenting cell from which the exosome is derived. Empty MHC molecules on exosomes may also be loaded with peptides afterwards.

Abstract: The present invention provides a genomic DNA of Buchnera. That is, this invention provides genes derived from Buchnera sp., comprising DNA of the following (a) or (b); (a) a DNA selected from a group consisting of a DNA having a nucleotide sequence ranging from a start point to an end point as shown in Table 1 in a nucleotide sequence represented by SEQ ID NO:1, or a DNA complementary thereto, and (b) a DNA hybridizing to the DNA of (a) under stringent conditions and encoding a protein having a function same as that of the product expressed by the DNA.

Abstract: This invention relates to a process by which an extract comprising integral thylakoids is obtained. The resulting extract is a potent dynamic antioxidant useful as a ROS (reactive oxygen species) scavenger. This extract is intended to be used for the treatment or prevention of diseases involving the generation of ROS, such as inflammatory diseases or cancer. This extract also finds a use as a solar screen because of its capacity to capture UV radiations and to dissipate the solar energy into heat.

Abstract: The present invention provides chimeric polypeptides and nucleic acids encoding the chimeric polypeptides and methods for using the same to detect and measure protease activity. It further provides in vivo and in vitro methods for identifying modulators of protease activity, e.g., by high throughput assays.

Abstract: The present invention is directed to methods for producing virus-free vesicles containing functional membrane proteins, and uses thereof. Preferably the virus-free vesicles contain a type II transmembrane protein which is a member of the TNF superfamily. The vesicles maintain the protein in its native conformation, thus eliminating the problems related to the solubility of these proteins when produced without presence of membrane component.