Abstract: We have cloned human lymphoid cell lines that are susceptible to hepatitis C virus (HCV) infection, and in which infection with HCV results in the development of cytopathic effects, including cell degeneration, induction of cell syncytia and cell death, as well as in the production of progeny virus. Infection was confirmed by the polymerase chain reaction, indirect immunofluorescence of viral antigens, and detection of the viral RNA-dependent RNA polymerase. Progeny virus released from the infected cells into the medium could be serially passaged using the cell-free supernatant fluid as the inoculum. Also described are uses of the cloned cell lines for both intact cell and cell-free assay systems for the effectiveness of candidate anti-HCV drugs.
Abstract: Computer implemented processes are disclosed for quantifying the severity of condition of patients of a pediatric hospital or pediatric intensive care unit. The measure of severity of condition can be used to calculate a probability of death during hospitalization as well as the expected length of stay for a patient, given information gathered about the patient during the initial hours of hospitalization. This information may then be used to determine availability of beds and to provide objective measures of the quality of care provided by a pediatric hospital or part thereof.
Abstract: A new homogeneous cytosolic binding protein (FKBP-14.6 immunophilin) having a molecular weight of about 14.6 kDa (calculated from its amino acid composition), reversibly binds the immunosuppressive drugs FK-506, rapamycin or chemically related compounds, but not cyclosporine. The FKBP-14.6 protein has a pI of 6.5-7.5, and the (partial) N-terminal amino acid sequence: NH.sub.2 -Lys-Leu-Pro-Tyr-Glu-Leu-Lys-X-Asn-Val-Lys-Ala-Phe-X-X-Lys-Val- (where X is undefined), and partial internal amino acid sequences -Val-Leu-Asp-Thr-Ala-Tyr-Glu-Tyr-Gly-Ala-Glu-Ala-Leu-Glu-, -Glu-Phe-Thr-Pro-Val-Phe-Gln-Ala-X-Phe- and -Ser-Leu-Val-Pro-Leu-Val-Gly-X-Lys- (where X is undefined). This N-terminal amino acid sequence exhibits no homology to FKBP-12 or membrane-associated FKBP-13. THe FKBP-14.6 is isolated from the cytosol of mammalian tissues, preferably calf thymus, and can be used in diagnostic and purification procedures involving FK-506 and rapamycin-type immunosuppressant drugs.
Abstract: A new homogeneous cytosolic binding protein (FKBP-14.6 immunophilin) having a molecular weight of about 14.6 kDa (calculated from its amino acid composition), reversibly binds the immunosuppressive drugs FK-506, rapamycin or chemically related compounds, but not cyclosporine. The FKBP-14.6 protein has a pI of 6.5-7.5, and the (partial) N-terminal amino acid sequence: NH.sub.2 -Lys-Leu-Pro-Tyr-Glu-Leu-Lys-Xaa-Asn-Val-Lys-Ala-Phe-Xaa-Xaa-Lys-Val- (Seq. ID. No: 1) (where Xaa is undefined), and partial internal amino acid sequences -Val-Leu-Asp-Thr-Ala-Tyr-Glu-Tyr-Gly-Ala-Glu-Ala-Leu-Glu- (Seq. ID. No: 2), -Glu-Phe-Thr-Pro-Val-Phe-Gln-Ala-Xaa-Phe- (Seq. ID. No: 3) and -Ser-Leu-Val-Pro-Leu-Val-Gly-Xaa-Lys- (Seq. ID. No. 4) (where Xaa is undefined). This N-terminal amino acid sequence exhibits no homology to FKBP-12 or membrane-associated FKBP-13. The FKBP-14.
Abstract: A competitive protein binding assay for rapamycin and biologically-active metabolites, derivatives and analogues thereof in blood and other biological fluids is disclosed, wherein the binding reagent is a specific rapamycin binding protein either substantially purified from the soluble cytoplasm of target cells of rapamycin action, particularly normal or transformed lymphocytes, freshly collected or in established cell lines, or synthesized by recombinant DNA techniques. Solution phase and solid state assay systems are disclosed.