Abstract: Novel cell penetrating agents for intracellular delivery of desired cargo, including proteins. Use of cell penetrating agents to deliver cargos to the interior of cells and cellular compartments and organelles is transformative for diagnostic, therapeutic, and research processes.

Abstract: A bearing housing assembly featuring a bearing housing having a bearing housing wall portion with a bearing assembly chamber for receiving a bearing assembly and a shaft to be rotated, an oil sump for receiving and containing oil for lubricating the bearing assembly when the shaft is rotated, an oil path channel formed as an oil path for receiving dirty oil from the bearing assembly chamber for traveling down the oil path, and a filter assembly wall portion forming a filter assembly cavity coupled fluidically between the oil sump and the oil path channel; and a filter assembly arranged in the filter assembly cavity, to couple to the filter assembly wall portion, receive the dirty oil traveling down the oil path, filter the dirty oil and provide filtered oil to the oil sump, so the filtered oil can be recirculated to lubricate the bearing assembly when the shaft is rotated.

Abstract: Novel cell penetrating agents for intracellular delivery of desired cargo, including proteins. Use of cell penetrating agents to deliver cargos to the interior of cells and cellular compartments and organelles is transformative for diagnostic, therapeutic, and research processes.

Abstract: A bearing housing assembly featuring a bearing housing having a bearing housing wall portion with a bearing assembly chamber for receiving a bearing assembly and a shaft to be rotated, an oil sump for receiving and containing oil for lubricating the bearing assembly when the shaft is rotated, an oil path channel formed as an oil path for receiving dirty oil from the bearing assembly chamber for traveling down the oil path, and a filter assembly wall portion forming a filter assembly cavity coupled fluidically between the oil sump and the oil path channel; and a filter assembly arranged in the filter assembly cavity, to couple to the filter assembly wall portion, receive the dirty oil traveling down the oil path, filter the dirty oil and provide filtered oil to the oil sump, so the filtered oil can be recirculated to lubricate the bearing assembly when the shaft is rotated.

Abstract: Novel cell penetrating agents for intracellular delivery of desired cargo, including proteins. Use of cell penetrating agents to deliver cargos to the interior of cells and cellular compartments and organelles is transformative for diagnostic, therapeutic, and research processes.

Abstract: A bearing housing assembly featuring a bearing housing having a bearing housing wall portion with a bearing assembly chamber for receiving a bearing assembly and a shaft to be rotated, an oil sump for receiving and containing oil for lubricating the bearing assembly when the shaft is rotated, an oil path channel formed as an oil path for receiving dirty oil from the bearing assembly chamber for traveling down the oil path, and a filter assembly wall portion forming a filter assembly cavity coupled fluidically between the oil sump and the oil path channel; and a filter assembly arranged in the filter assembly cavity, to couple to the filter assembly wall portion, receive the dirty oil traveling down the oil path, filter the dirty oil and provide filtered oil to the oil sump, so the filtered oil can be recirculated to lubricate the bearing assembly when the shaft is rotated.

Abstract: Cell penetrating peptides (CPPs) are established as a strategy to move cargoes into the interior of eukaryotic cells by engaging import machinery on the cell surface. In most cases the CPP is covalently linked to the cargo; it is common to express cargo proteins with a CPP extension. In some experiments a non-specific interactions (e.g., hydrophobic interactions) have been used to form CPP-cargo complexes, but this has numerous drawbacks. Transport is less efficient and the lack of specificity means that other macromolecules in the medium will also be internalized. This application describes the use of specific CPP labeled adaptor proteins that can be used to move a wide variety of cargoes into the cell interior. The prototype adaptor protein is calmodulin; it is small, stable and easily produced, and binds short (17 amino acid) targets with high affinity in the presence of calcium. A cargo produced with a calmodulin binding tag can be internalized efficiently by CPP tagged calmodulin.

Abstract: Cell penetrating peptides (CPPs) are established as a strategy to move cargoes into the interior of eukaryotic cells by engaging import machinery on the cell surface. In most cases the CPP is covalently linked to the cargo; it is common to express cargo proteins with a CPP extension. In some experiments a non-specific interactions (e.g., hydrophobic interactions) have been used to form CPP-cargo complexes, but this has numerous drawbacks. Transport is less efficient and the lack of specificity means that other macromolecules in the medium will also be internalized. This application describes the use of specific CPP labeled adaptor proteins that can be used to move a wide variety of cargoes into the cell interior. The prototype adaptor protein is calmodulin; it is small, stable and easily produced, and binds short (17 amino acid) targets with high affinity in the presence of calcium. A cargo produced with a calmodulin binding tag can be internalized efficiently by CPP tagged calmodulin.

Abstract: Coupling proteins that make strong protein-protein interactions equipped with cell penetrating peptides (CPPs) provide a convenient and powerful method to perturb cell interiors; there are many potential payloads and a broad palette of selectively membrane permeable probes. In a preferred embodiment, the coupling protein will be calmodulin or a related calcium binding protein. In a preferred embodiment, the CPP will be TAT or another CPP. In a preferred embodiment, the coupling protein will release its payload after targeting to an interior compartment. Cargo proteins can be purified by affinity methods using the same tag that allows binding by the adaptor, enabling an integrated approach with ‘gains in both function and safety. Access to cell interior compartments has potential applications in research, diagnostics, and therapeutics.

Abstract: Novel cell penetrating agents for intracellular delivery of desired cargo, including proteins. Use of cell penetrating agents to deliver cargos to the interior of cells and cellular compartments and organelles is transformative for diagnostic, therapeutic, and research processes.

Abstract: A bearing housing assembly featuring a bearing housing having a bearing housing wall portion with a bearing assembly chamber for receiving a bearing assembly and a shaft to be rotated, an oil sump for receiving and containing oil for lubricating the bearing assembly when the shaft is rotated, an oil path channel formed as an oil path for receiving dirty oil from the bearing assembly chamber for traveling down the oil path, and a filter assembly wall portion forming a filter assembly cavity coupled fluidically between the oil sump and the oil path channel; and a filter assembly arranged in the filter assembly cavity, to couple to the filter assembly wall portion, receive the dirty oil traveling down the oil path, filter the dirty oil and provide filtered oil to the oil sump, so the filtered oil can be recirculated to lubricate the bearing assembly when the shaft is rotated.

Abstract: Coupling proteins that make strong protein-protein interactions equipped with cell penetrating peptides (CPPs) provide a convenient and powerful method to perturb cell interiors; there are many potential payloads and a broad palette of selectively membrane permeable probes. In a preferred embodiment, the coupling protein will be calmodulin or a related calcium binding protein. In a preferred embodiment, the CPP will be TAT or another CPP. In a preferred embodiment, the coupling protein will release its payload after targeting to an interior compartment. Cargo proteins can be purified by affinity methods using the same tag that allows binding by the adaptor, enabling an integrated approach with ‘gains in both function and safety. Access to cell interior compartments has potential applications in research, diagnostics, and therapeutics.

Abstract: The invention provides improved mutagenic plasmids and a method for making and using them. In one embodiment a cyclic mutagenic nucleic acid is replicated to form interlocking, digestibly separable duplex or simplex rings; and then one or more of the interlocked rings are cleaved selectively by means of a restriction enzyme to liberate an intact duplex or simplex ring. The mutagenic plasmids show substantially improved efficiencies in transforming cellular microorganism that are exposed to them.

Abstract: Cell penetrating peptides (CPPs) are established as a strategy to move cargoes into the interior of eukaryotic cells by engaging import machinery on the cell surface. In most cases the CPP is covalently linked to the cargo; it is common to express cargo proteins with a CPP extension. In some experiments a non-specific interactions (e.g., hydrophobic interactions) have been used to form CPP-cargo complexes, but this has numerous drawbacks. Transport is less efficient and the lack of specificity means that other macromolecules in the medium will also be internalized. This application describes the use of specific CPP labeled adaptor proteins that can be used to move a wide variety of cargoes into the cell interior. The prototype adaptor protein is calmodulin; it is small, stable and easily produced, and binds short (17 amino acid) targets with high affinity in the presence of calcium. A cargo produced with a calmodulin binding tag can be internalized efficiently by CPP tagged calmodulin.

Abstract: Endothelial nitric oxide synthase (eNOS) activity was modulated by contact with an effective amount of a mitogen-activated protein (MAP) kinase, resulting in phosphorylation of S602, T46, and/or S58. Kinetics and stoichiometry are disclosed. The contact strongly reduced nitric oxide (NO) synthesis, and inhibited the cytochrome c reductase activity of eNOS reductase domains. Three sites of phosphorylation were determined that matched the serine-proline (SP) and threonine-proline (TP) motifs of typical MAP kinase phosphorylation sites.

Abstract: The invention provides improved mutagenic plasmids and a method for making and using them. In one embodiment a cyclic mutagenic nucleic acid is replicated to form interlocking, digestibly separable duplex or simplex rings; and then one or more of the interlocked rings are cleaved selectively by means of a restriction enzyme to liberate an intact duplex or simplex ring. The mutagenic plasmids show substantially improved efficiencies in transforming cellular microorganism that are exposed to them.

Abstract: The activity of endothelial nitric oxide synthase (eNOS) was modulated by contact with an effective amount of a mitogen-activated protein (MAP) kinase, resulting in phosphorylation of S602, T46, and/or S58. Kinetics and stoichiometry are disclosed. The contact strongly reduced nitric oxide (NO) synthesis, and also inhibited the cytochrome c reductase activity of eNOS reductase domains. Three sites of phosphorylation were determined that matched the serine-proline (SP) and threonine-proline (TP) motifs of typical MAP kinase phosphorylation sites.

Abstract: Methods for generating chimeric polynucleotides by directed evolution are described. In the methods, splice points of interest are identified within the polynucleotides of a basis set of polynucleotides, preferably through the use of an algorithm that defines the number of splice points and selects the splice points, either by random selection or using information regarding alignment of the polynucleotides. The algorithms can include additional factors, including a definition of a desired distance between splice points, and/or weighing factors to bias selection of splice points. Chimeric polynucleotides are generated using primers (e.g., double primers or non-overlapping primers) and polymerase chain reaction or combinatorial strategies.

Abstract: The present invention relates to a novel protein expression system having an oligonucleotide encoding a small heat shock protein (sHSP) operably linked to a promoter and an oligonucleotide encoding a protein of interest. In one embodiment the expressed sHSP is a truncated &agr;-crystallin polypeptide derived from a wild-type &agr;-crystallin protein, wherein the truncated sHSP lacks an N-terminal sequence present in the wild-type &agr;-crystallin polypeptide. In an additional embodiment, a protein is coexpressed with a sHSP, thereby increasing the level of expression, enhancing folding and increasing the solubility of the protein.