Abstract: Methods for transforming Pichia methanolica, and DNA molecules useful in transformation of P. methanolica, are disclosed. P. methanolica cells are exposed, in the presence of DNA molecules, to a pulsed electric field having a field strength of from 2.5 to 4.5 kV/cm and a pulse duration of from 1 to 40 milliseconds, whereby the DNA molecules are introduced into the cells. The DNA molecules may comprise an expression unit that includes a transcription promoter of a P. methanolica gene operably linked to a segment encoding a polypeptide or protein of interest. The DNA molecules may also encode a selectable marker, such as a P. methanolica ADE2 gene. Cells transformed according to the invention may be used in production systems for the preparation of proteins of commercial importance.

Abstract: Enzymatically cross-linked protein gels and methods for preparing them are disclosed. The methods comprise adding a transglutaminase, such as factor XIII, to a composition of a temperature-sensitive gel-forming protein, such as gelatin or collagen, and incubating the composition and transglutaminase under gel-forming conditions. The resulting gels have superior strength and thermal stability, and can be used within a variety of medical and industrial applications.

Abstract: Novel receptor polypeptides, polynucleotides encoding the polypeptides, and related compositions and methods are disclosed. The polypeptides comprise an extracellular ligand-binding domain of a cell-surface receptor that is expressed at high levels in lymphoid tissue, including B-cells and T-cells. The polypeptides may be used within methods for detecting ligands that stimulate the proliferation and/or development of lymphoid and myeloid cells in vitro and in vivo. Ligand-binding receptor polypeptides can also be used to block ligand activity in vitro and in vivo.

Abstract: Methods of removing protein contaminants from a solution comprising thrombopoietin are disclosed. The solution is exposed to hydroxyapatite, whereby protein contaminants are bound to the hydroxyapatite and the thrombopoietin remains substantially unbound, and the unbound thrombopoietin is recovered. The use of hydroxyapatite is advantageously combined with other purification and concentration techniques, including ion-exchange chromatography, ligand affinity chromatography, hydrophobic interaction chromatography, ultrafiltration, and differential precipitation.

Abstract: Isolated DNA molecules comprising a portion of a human hexokinase gene promoter are disclosed. The molecules comprise promoter and insulin-sensitive transcription regulatory elements. These molecules can be used within methods for detecting insulin-like activity in test substances.

Abstract: Methods for preparing Pichia methanolica cells having auxotrophic mutations are disclosed. The methods comprise the steps of (a) exposing P. methanolica cells to mutagenizing conditions, (b) culturing the cells from step (a) in a rich medium to allow mutations to become established and replicated in at least a portion of the cells, (c) culturing the cells from step (b) in a culture medium deficient in assimilable nitrogen to deplete cellular nitrogen stores, (d) culturing the cells from step (c) in a defined culture medium comprising an inorganic nitrogen source and an amount of nystatin sufficient to kill growing P. methanolica cells to select for cells having a deficiency in a nutritional gene; and (e) culturing the selected cells from step (d) in a rich culture medium.

Abstract: Methods are provided for producing protein products in host cells and for selecting transformed cells comprising the step of transforming the host cell with a DNA molecule comprising a gene which complements a deficiency in the host cell. The host cell is s strain having a deficiency in a function necessary for normal cell growth. The gene in the DNA molecule, such as a plasmid, which complements the deficiency serves as a selection marker whereby the growth conditions for selection may comprise a conventional complex medium.

Abstract: Methods for preparing Pichia methanolica cells containing foreign DNA constructs and methods for producing foreign polypeptides in Pichia methanolica cells are disclosed. The cells are transformed with a DNA construct comprising a first DNA segment comprising a transcription promoter of a methanol-inducible P. methanolica gene, a second DNA segment encoding a higher eukaryotic polypeptide, a third DNA segment comprising a P. methanolica gene transcription terminator, and a selectable marker. For production of foreign polypeptides, the cells are cultured under conditions in which the second DNA segment is expressed, and the polypeptide encoded by the second DNA segment is recovered.

Abstract: Methods for obtaining cells that produce a ligand for an orphan receptor and methods for preparing polynucleotide molecules that encode ligands for orphan receptors are disclosed. The methods utilize growth factor-dependent parent cells that are transfected with a DNA construct encoding an orphan receptor. The transfected cells are exposed to mutagenizing conditions, and the mutagenized cells are cultured under conditions in which cell survival is dependent upon autocrine growth factor production. Progeny cells are recovered and screened to identify those that produce a ligand for the orphan receptor. Polynucleotide molecules encoding the ligand can be prepared from the identified cells.

Abstract: Methods are provided for producing protein products in host cells and for selecting transformed cells comprising the step of transforming the host cell with a DNA molecule comprising a gene which complements a deficiency in the host cell. The host cell is a strain having a deficiency in a function necessary for normal cell growth. The gene in the DNA molecule, such as a plasmid, which complements the deficiency serves as a selection marker whereby the growth conditions for selection may comprise a conventional complex medium.

Abstract: The present invention provides an immature dendritic cell line derived from p53 growth suppressor gene deficient animals. The immature dendritic cell line may be induced to become an activated dendritic cell line that will stimulate T-cells to proliferate. The cell line is useful for presentation of antigens involved in autoimmune disease and analysis of peptides that produce a T-cell response.

Abstract: The present invention relates to an orthogonally-protected compound of the formula: ##STR1## wherein PG.sub.1 is a first protecting group that is unreactive to a chemistry used at another position within the compound, and that is capable of selective removal without affecting PG.sub.2 or linkage to a solid support; PG.sub.2 is a second protecting group that is unreactive to a chemistry used at another position within the compound, and that is capable of selective removal without affecting PG.sub.1 or linkage to a solid support; Y is CH.sub.2 COOH, CH.sub.2 SO.sub.2 OH, CH.sub.2 PO.sub.2 ROH, CH.sub.2 Ph--COOH, CH.sub.2 Ph--SO.sub.2 OH, or CH.sub.2 Ph--PO.sub.2 ROH; R is H or a substituted or unsubstituted alkyl, cycloalkyl, aryl, arylalkyl, heteroaryl, or heteroarylalkyl; and n is 1 or 2. PG.sub.1 is preferably a carbamate group, a trityl group, or a trifluoroacetyl group; and PG.sub.2 is preferably an ester.

Abstract: DNA constructs useful in the production of thrombopoietin are disclosed. In general, the DNA constructs comprise a first DNA segment encoding a fusion of an amino-terminal secretory peptide joined to a thrombopoietin polypeptide and one or more additional DNA segments that provide for the transcription of the first segment. The secretory peptide is a native mammalian t-PA secretory peptide or may be modified to enhance proteolytic cleavage of the fusion. Also disclosed are cultured eukaryotic cells containing these DNA constructs and methods for producing thrombopoietin polypeptides through the use of the DNA constructs and cultured eukaryotic cells.

Abstract: Materials and methods for producing fibrinogen in transgenic non-human mammals are disclosed. DNA segments encoding A.alpha., B.beta. and .gamma. chains of fibrinogen are introduced into the germ line of a non-human mammal, and the mammal or its female progeny produces milk containing fibrinogen expressed from the introduced DNA segments. Non-human mammalian embryos and transgenic non-human mammals carrying DNA segments encoding heterologous fibrinogen polypeptide chains are also disclosed.

Abstract: Isolated polynucleotide molecules encoding mammalian phospholipid transfer proteins (PLTP) and phospholipid transfer protein polypeptides are disclosed. The DNA molecules are transformed or transfected into host cells and the cells cultured to produce recombinant PLTP and PLTP polypeptides. PLTP and PLTP polypeptides may be combined with a pharmaceutically acceptable vehicle and administered to patients to regulate phospholipid transfer activity and thereby obtain a more favorable lipoprotein profile in the blood. The proteins and polypeptides may also be used within methods to measure phospholipid transfer activity or identify inhibitors of phospholipid transfer activity.

Abstract: Methods for inhibiting intimal hyperplasia in the vasculature of mammals, including primates, are disclosed. The methods comprise administering to the mammal an anti-PDGF receptor antibody, such as an anti-PDGF-alpha receptor antibody or an anti-PDGF-beta receptor antibody. The methods are useful in reducing intimal hyperplasia due to, for example, vascular injuries resulting from angioplasty, endarterectomy, reduction atherectomy or anastomosis of a vascular graft. The anti-PDGF receptor antibodies may optionally be administered coordinately with heparin, whereby the coordinately administered antibody and heparin are combinatorially effective in inhibiting intimal hyperplasia.

Abstract: Methods for inhibiting intimal hyperplasia in the vasculature of mammals, including primates, are disclosed. The methods comprise administering to the mammal an effective amount of Brefeldin A or a derivative of Brefeldin A. The methods are useful in reducing intimal hyperplasia due to, for example, vascular injuries resulting from angioplasty, endarterectomy, reduction atherectomy or anastomosis of a vascular graft. The non-peptide PDGF antagonists Brefeldin A and its derivatives may optionally be administered coordinately with heparin, whereby the coordinately administered of non-peptide PDGF antagonist and heparin are combinatorially effective in inhibiting intimal hyperplasia.

Abstract: Isolated polynucleotide molecules encoding mammalian phospholipid transfer proteins (PLTP) and phospholipid transfer protein polypeptides are disclosed. The DNA molecules are transformed or transfected into host cells and the cells cultured to produce recombinant PLTP and PLTP polypeptides. PLTP and PLTP polypeptides may be combined with a pharmaceutically acceptable vehicle and administered to patients to regulate phospholipid transfer activity and thereby obtain a more favorable lipoprotein profile in the blood. The proteins and polypeptides may also be used within methods to measure phospholipid transfer activity or identify inhibitors of phospholipid transfer activity.

Abstract: Blood loss in a patient undergoing surgery, particularly thoracic or abdominal surgery, is reduced by administration of factor XIII. The factor XIII may be administered in combination with aprotinin.

Abstract: Methods and pharmaceutical compositions for use in inhibiting calmodulin activity in a patient are disclosed. 2,3-diaryl-1-benzopyrans and their pharmaceutically acceptable salts are formulated into medicaments, including oral and topical medicaments, which are administered to a patient in need of treatment of a non-estrogen dependent condition.