Abstract: Methods and compositions for detecting molecular interactions, particularly protein-protein interactions, using at least two inactive, weakly-complementing ?-lactamase fragments are provided. The invention allows detection of such interactions in eukaryotic and mammalian cells, in situ or in vitro. Detection of molecular interactions in mammalian cells is not limited to the nuclear compartment, but can be accomplished in the cytoplasm, cell surface, organelles, or between these entities. Methods provided utilize novel compositions comprising fusion proteins between molecules of interest and inactive, weakly-complementing ?-lactamase fragments. Association of the molecules of interest brings the corresponding complementary ?-lactamase fragments into close enough proximity for complementation to occur and ?-lactamase activity to be observed. The invention is useful in the study of protein-protein interactions, functional genomics, agonist and antagonist screening and drug discovery.

Abstract: Methods and compositions are provided for detecting molecular translocations, particularly protein translocations within and between sub-cellular compartments, using at least two components that exhibit a localization-dependent difference in complementation activity. In particular, alpha-complementing ?-galactosidase fragments are provided. These ?-galactosidase reporter fragments display significantly enhanced enzymatic activity when one fragment is localized in a membrane. Methods for carrying out no-wash ELISA assays based on the reporter component system are also provided.

Abstract: Provided is a sequenced fluid control device that includes a pneumatic driver module, a fluid cartridge having at least one fluid chamber and at least one waste chamber, and at least one sample delivery element disposed to sealably contact the fluid cartridge forming a main chamber containing a sample, where the fluid cartridge is also disposed to sealably contact the pneumatic driver module. The pneumatic driver module injects gas into or withdraws gas from the fluid cartridge, where the gas directly contacts at least one fluid causing at least one fluid to flow through channels disposed to connect at least one fluid chamber to the sample delivery element and disposed to connect the sample delivery element to the at least one waste chamber or waste channel, where a sample on the sample delivery element is exposed to the at least one fluid.

Abstract: Methods and compositions for detecting the sub-cellular localization of a molecule are provided. Aspects of the invention include detecting translocation of a cell-surface receptor to a sub-cellular compartment, e.g., the endosome, using a reduced affinity enzyme complementation reporter system. Also provided are systems and kits for use in practicing embodiments of the methods.

Abstract: The embodiments of the present disclosure encompass methods for non-invasive in vivo bioluminescence imaging that allow the dynamics of stem cell behavior to be followed in a manner not possible using conventional retrospective static histological analyses. By imaging luciferase-generated bioluminescence activity emanating from isolated stem cells, for example, real time quantitative and kinetic analyses can show that donor-derived muscle stem cells may proliferate and engraft rapidly after injection until homeostasis is reached. In addition, the response of the stem cells to injury and participation in the regenerative response can be monitored over time. Other aspects of the disclosure encompasses methods for determining the suitability of a stem cell for tissue replacement, methods for repairing muscle injury, and methods for isolating muscle stem cells from a tissue sample.

Abstract: Embodiments of the present disclosure encompass microfabrication methods (“reactive microcontact printing of soft matter”) for hydrated soft polymer materials and surfaces for culture platforms suitable for the culturing of isolated single primary mammalian cells in an environment approximating the natural niches of the cells. Such culture platforms may comprise arrays of microwells, or other microscopically textured features, in which individual features can comprise desired proteins or mixtures of proteins. The microfabrication methods of the disclosure allow spatial control of surface biochemistry and topography at the micrometer scale on these hydrated soft gels. The hydrogels and methods of manufacture and use of the disclosure allow the isolation of a single stem cell and the characterizing of its interaction with cytokines and morphogens, especially with regard to modulation of the proliferative capacity of the stem cell when implanted in a recipient host.

Abstract: The disclosure encompasses transparent polymer membranes suitable for use in laser microdissection methods, where the membranes allow the subsequent analysis of the microdissected cell or cells with light of a wavelength that can traverse the membrane with little if any absorbance by the membrane. The transfer capture membranes of the disclosure comprise a polymeric composition having the characteristic of being cut by a laser light at a wavelength less than 360 nm, while being substantially optically transparent at a wavelength greater than about 360 nm.

Abstract: Sequential reporter enzyme luminescence (SRL) methods are provided. In the subject methods, the activity of a reporter enzyme is evaluated using a secondary reporter system that employs a product of a reporter enzyme mediated reaction as a luminescent substrate, e.g., luciferase substrate. Also provided are kits and other compositions that find use in practicing the subject methods.

Abstract: Methods and compositions for detecting molecular interactions, particularly protein-protein interactions, are provided. The invention allows detection of such interactions in living cells or in vitro. Detection of molecular interactions in living cells is not limited to the nuclear compartment, but can be accomplished in the cytoplasm, cell surface, organelles, or between these entities. In one embodiment, the method utilizes novel compositions comprising fusion proteins between the molecules of interest and two or more inactive, weakly-complementing ?-galactosidase mutants. Association between the molecules of interest brings the complementing ?-galactosidase mutants into proximity so that complementation occurs and active ?-galactosidase is produced. The active ?-galactosidase may be detected by methods well-known in the art. Among the uses of the invention are the study of protein-protein interactions, functional genomics, agonist and antagonist screening and drug discovery.

Abstract: The invention provides, among other things, novel methods of treating neurological disorders which result in the loss of neurons (neuronal deficiencies). Bone marrow-derived cells are administered to individuals suffering from neuronal deficiencies. Administration of bone marrow-derived cells results in formation of bone marrow derived neurons, whether formed de novo or as a result of fusion with an existing neuron, thereby replacing or repairing lost or damaged neurons. The methods of the invention may also be used for memory augmentation in memory impaired individuals.

Abstract: Methods and compositions for detecting molecular interactions, particularly protein-protein interactions, using at least two inactive, weakly-complementing &bgr;-lactamase fragments are provided. The invention allows detection of such interactions in eukaryotic and mammalian cells, in situ or in vitro. Detection of molecular interactions in mammalian cells is not limited to the nuclear compartment, but can be accomplished in the cytoplasm, cell surface, organelles, or between these entities. Methods provided utilize novel compositions comprising fusion proteins between molecules of interest and inactive, weakly-complementing &bgr;-lactamase fragments. Association of the molecules of interest brings the corresponding complementary &bgr;-lactamase fragments into close enough proximity for complementation to occur and &bgr;-lactamase activity to be observed. The invention is useful in the study of protein-protein interactions, functional genomics, agonist and antagonist screening and drug discovery.

Abstract: The invention provides novel methods of treating neurological disorders which result in the loss of neurons (neuronal deficiencies). Bone marrow-derived cells are administered to individuals suffering from neuronal deficiencies. Administration of bone marrow-derived cells results in formation of new neurons in the nervous system, thereby replacing lost neurons. The methods of the invention may also be used for memory augmentation in memory impaired individuals.

Abstract: Methods and compositions for detecting molecular interactions, particularly protein-protein interactions, are provided. The invention allows detection of such interactions in living cells or in vitro. Detection of molecular interactions in living cells is not limited to the nuclear compartment, but can be accomplished in the cytoplasm, cell surface, organelles, or between these entities. In one embodiment, the method utilizes novel compositions comprising fusion proteins between the molecules of interest and two or more inactive, weakly-complementing &bgr;-galactosidase mutants. Association between the molecules of interest brings the complementing &bgr;-galactosidase mutants into proximity so that complementation occurs and active &bgr;-galactosidase is produced. The active &bgr;-galactosidase may be detected by methods well-known in the art. Among the uses of the invention are the study of protein-protein interactions, functional genomics, agonist and antagonist screening and drug discovery.

Abstract: Methods and compositions for detecting molecular interactions, particularly protein-protein interactions, are provided. The invention allows detection of such interactions in living cells or in vitro. Detection of molecular interactions in living cells is not limited to the nuclear compartment, but can be accomplished in the cytoplasm, cell surface, organelles, or between these entities. In one embodiment, the method utilizes novel compositions comprising fusion proteins between the molecules of interest and two or more inactive, weakly-complementing &bgr;-galactosidase mutants. Association between the molecules of interest brings the complementing &bgr;-galactosidase mutants into proximity so that complementation occurs and active &bgr;-galactosidase is produced. The active &bgr;-galactosidase may be detected by methods well-known in the art. Among the uses of the invention are the study of protein-protein interactions, functional genomics, agonist and antagonist screening and drug discovery.

Abstract: Myoblasts are produced, conveniently in low or serum-free medium, for use in introduction into a mammalian host, particularly a human host, for treatment of diseases of muscle tissue or acting as carriers for genetic capabilities, particularly correcting a genetic defect or for production of a soluble protein, which may serve in a therapy for the mammalian host. Myoblasts introduced into tissue are able to migrate to sites distal from the site of injection, expanding the area of their effect.