Abstract: Peptides derived from three regions of the lectin domain of GMP-140 (P-selectin) and the related selectins, ELAM-1 (E-selectin) and the lymphocyte homing receptor (L-selectin), have been found to inhibit neutrophil adhesion to GMP-140. These and additional peptides have been synthesized, having as their core region portions of the 74-76 amino acid sequence of GMP-140, with residue 1 defined as the N-terminus of the mature protein after the cleavage of the signal peptide. Examples demonstrate the inhibition of the binding of neutrophils to GMP-140 of peptides in concentrations ranging from 30 to 1500 .mu.mol. It has been found that alterations within the core sequence, as well as N-terminal and C-terminal flanking regions, do not result in loss of biological activity. The peptides are useful as diagnostics and, in combination with a suitable pharmaceutical carrier, for clinical applications in the modulation or inhibition of coagulation processes or inflammatory processes.

Abstract: Complementary DNA encoding a 52 kDa form of a protein present in the human Ro/SSA ribonucleoprotein complex has been cloned. A lambda gt11 cDNA library made from human thymocyte mRNA was screened with serum from a SLE patient and two immunoreactive clones were isolated. These clones reacted with other patient sera which had anti-52 kDa Ro/SSA antibodies and with affinity purified anti-52 kDa Ro/SSA antibodies. Moreover, affinity purified antibodies eluted from fusion proteins of the isolated clones reacted only with the 52 kDa protein of lymphocytes in the Western blot. Ro/SSA RNAs were also precipitated with these affinity purified antibodies, further confirming that the clones encode a 52 kDa Ro/SSA antigen. The sequence differs from the previously reported 60 kDa Ro/SSA gene.

Abstract: A method of detecting dysplastic regions within epithelial tissue samples is sensitive enough to detect and distinguish between low grade and high grade dysplastic regions. The method uses probes specific for expression and accumulation of substances within a particular intracellular region from a defect in apical membrane trafficking (trafficking markers) and in the preferred embodiment correlates the trafficking marker levels with the presence of an oncogene such as p53. If low grade dysplasia is present, trafficking markers are detected in a distinctive perinuclear pattern. Previous studies have demonstrated a high correlation of p53 over-expression with high grade dysplasia and adenocarcinoma. Detection of p53 is shown to be mutually exclusive of detection of trafficking markers. Therefore, dual detection for both the trafficking markers and p53 provides an accurate method for more precise grading of biopsies.

Abstract: Compositions and methods using antibodies which are immunoreactive with specific apolipoproteins to determine the concentrations of lipoproteins such as HDL and LDL, and/or apolipoproteins in human blood, serum or plasma sample, are described. Monoclonal antibodies (MAbs) are described that specifically bind to epitopes present in apolipoproteins and lipoproteins, enabling rapid and reliable determinations of levels of specific blood lipoprotein and/or apolipoprotein levels, including Apo B-100, Apo A-I, Apo A-II, Apo C-III, and Apo E, and thereby determination of relative ratios of HDL and LDL and LpaI and LpaII. In a preferred embodiment, the compositions are strips of a solid phase material coated with one or more of the antibodies and are referred to herein as "dipsticks". The dipsticks specifically bind a lipoprotein or apolipoprotein when dipped into a protein sample.

Abstract: Genetically engineered cells are provided which can serve as universal donor cells in such applications as reconstruction of vascular linings or the administration of therapeutic agents. The cells include a coding region which provides protection against complement-based lysis, i.e., hyperacute rejection. In addition, the cell's natural genome is changed so that functional proteins encoded by either the class II or both the class I and the class II major histocompatibility complex genes do not appear on the cell's surface. In this way, attack by T-cells is avoided. Optionally, the cells can include a self-destruction mechanism so that they can be removed from the host when no longer needed.

Abstract: A rapid, sensitive, and quantitative method and assay system using transgenic animal cells and adult or embryonic transgenic animals to rapidly screen compounds for mutagenic or teratogenic activity. Each cell from a cell culture, a differentiated tissue from an animal or an animal embryo contains a target gene and a reporter gene, both including an animal promoter/enhancer, a coding sequence and a transcription termination signal, so that they are capable of expression in animal cells. The product of the target gene is capable of regulating expression of the reporter gene through interaction with a regulatory sequence in the reporter gene. In the preferred embodiment, the target gene lacI (lac repressor) is coupled to the reporter gene lacZ encoding beta-galactosidase, which is easily detectable by cytochemical or histochemical procedures.

Abstract: It has been discovered that cells such as genetically engineered fibroblasts and keratinocytes can be cultured in the cyst fluid of encapsulated tumors. This provides a means for proliferating genetically engineered producer cells within these types of tumors, increasing the number of cells producing viral particles, which then transduce the surrounding tumor cells with the genetic material, in the preferred embodiment, a lethal gene. A number of different tumor types form "cysts", which contain fluid produced by the tumor cells, including brain tumor cells such as gliomas, and many types of breast, and lung tumors. These cyst fluids have been shown to contain elevated levels of certain growth factors, for example, fibroblast growth factor (FGF) and epidermal growth factor (EGF).

Abstract: The present invention describes a method for the production of two highly purified enzymes capable of degrading chondroitin sulfate polysaccharides. A multi-step purification method incorporating cell disruption, cation exchange chromatography, affinity chromatography, hydroxylapatite chromatography, high resolution ion exchange chromatography and size exclusion is outlined. A 77,000.+-.5,000 Dalton protein capable of degrading chondroitin sulfates A and C and a 55,000.+-.2,300 Dalton protein capable of degrading dermatan sulfate were isolated. The genes encoding these enzymes, chondroitinase AC and chondroitinase B, respectively, have been cloned and methods for their use are described.

Abstract: The present invention relates to a process for purifying apolipoprotein A (ApoA) or apolipoprotein E (ApoE) from human plasma, by obtaining a fraction of human plasma containing said ApoA or ApoE, prepurifying said fraction in at least one step, binding said ApoA or ApoE to an anion-exchange chromatography gel, and thereafter eluting said ApoA or ApoE from said anion-exchange chromatography gel. The thus produced ApoA or ApoE can be used for the manufacture of a medicament in the treatment of atherosclerosis and cardiovascular diseases, or peripheral atherosclerosis and sepsis as well as in a method for treatment of atherosderosis and cardiovascular diseases, or peripheral atherosclerosis and sepsis when administered in a therapeutically effective amount.

Abstract: A macromolecular delivery method that utilizes a series of peptides with unique and versatile nuclear targeting properties has been developed, where the peptides are derived from the COOH terminal domain (CTD) of the largest subunit of RNA polymerase II and include heptapeptide units similar or identical to the following consensus sequence: Tyrosine--Serine--Proline--Threonine--Serine--Proline--Serine (YSPTSPS).sub.x (SEQ ID NO. 1). When expressed in vivo, the CTD peptides are phosphorylated and they accumulate in discrete compartments within the nucleus. The CTD peptides concentrate indicator molecules in discrete subnuclear compartments where pre-mRNA molecules are synthesized and spliced. The length and composition of the CTD peptides can be manipulated to obtain different intranuclear partitioning properties. The CTD peptides are functional in the nuclei of S. cerevisiae, S. pombe, nematodes, insects, plants, and all vertebrates.

Abstract: The prevent invention relates to an integrated package-holder assay devices for detecting the presence of analyte in a sample. The device serves the dual roles of supporting and protecting an immunochromatographic assay. The device is compatible with any immunochromatographic assay format. The assay can be performed in a single apparatus for use in a laboratory or a field setting. In a specific example, the assay device is a nylon membrane formatted for an immunochromatographic assay for cotinine sealed between transparent adhesive tape and a stiff plastic strip. White tape placed over the plastic strip defined a window for observing the assay results.

Abstract: Methods are provided for separating polyhydroxyalkanoates ("PHAs") from plants, such as transgenic oil crop plants. The methods advantageously permit both the oil and the PHAs to be recovered from the plant biomass. To isolate the PHAs, in one embodiment, a biomass derived from an oil crop plant is pre-processed, for example by grinding, crushing or rolling. The oil then is extracted from the biomass with a first solvent in which the oil is soluble and in which the PHAs are not highly soluble to remove the oil. The biomass then can be extracted with a second solvent in which the PHA is soluble, to separate the PHA from the biomass. Alternatively, the PHA-containing biomass is treated with a chemical or biochemical agent, such as an enzyme, to chemically transform the PHA into a PHA derivative. The PHA derivative then is separated from the mixture using, for example, a physical separation process such as distillation, extraction or chromatography.

Abstract: Water-soluble macromers including at least one hydrolysable linkage formed from carbonate or dioxanone groups, at least one water-soluble polymeric block, and at least one polymerizable group, and methods of preparation and use thereof are described. The macromers are preferably polymerized using free radical initiators under the influence of long wavelength ultraviolet light or visible light excitation. Biodegradation occurs at the linkages within the extension oligomers and results in fragments which are non-toxic and easily removed from the body. The macromers can be used to encapsulate cells, deliver prophylactic, therapeutic or diagnostic agents in a controlled manner, plug leaks in tissue, prevent adhesion formation after surgical procedures, temporarily protect or separate tissue surfaces, and adhere or seal tissues together.

Abstract: Blood cholesterol levels are correlated with production of amyloid .beta. protein (A.beta.), and are predictors of populations at risk of developing AD. Methods for lowering blood cholesterol levels can be used to decrease production of A.beta., thereby decreasing the risk of developing AD. The same methods and compositions can also be used for treating individuals diagnosed with AD. Methods include administration of compounds which increase uptake of cholesterol by the liver, such as the administration of HMG CoA reductase inhibitors, administration of compounds which block endogenous cholesterol production, such as administration of HMG CoA reductase inhibitors, administration of compositions which prevent uptake of dietary cholesterol, and administration of combinations of any of these which are effective to lower blood cholesterol levels. Methods have also been developed to predict populations at risk, based on the role of cholesterol in production of A.beta..

Abstract: Biodegradable polymeric compositions are provided, wherein biodegradable polyphosphazenes are combined with at least one other polymer, either in the form of a blend, a semi-interpenetrating network (semi-IPN), or an interpenetrating network IPN. The side groups and composition of the polyphosphazenes are used to determine the properties of the compositions, for example, the rate and extent of degradation, and mechanical properties. These are useful in biomedical applications, including controlled drug delivery and tissue regeneration, and environmental applications. In the most preferred embodiment, as demonstrated by the examples, the polyphosphazenes contain hydrophobic side groups, such as p-methylphenoxy and other aromatic groups, and groups which impart hydrolytic instability, such as amino acid alkyl esters, and degrade by surface erosion. A preferred example is ethyl glycinato-substituted polyphosphazene (PPHOS) with p-methylphenoxy as co-substituent.

Abstract: A method for enhancing and/or increasing the efficiency of repair of tissues, primarily bone or cartilage, using genetically engineered cells has been developed. In the preferred embodiment, mesenchymal stem cells are isolated from periosteum tissue, and transfected with the gene encoding a growth factor for the particular cell type to be repaired. For example, for repair of bone, a gene (or genes) encoding bone morphogenic protein is transfected into periosteal cells. The transfected periosteal cells then express the bone morphogenic protein in culture to promote bone repair as a function of the expressed bone morphogenic protein. Cells can be transfected using any appropriate means, including viral vectors, as shown by the example, chemical transfectants, or physico-mechanical methods such as electroporation and direct diffusion of DNA.

Abstract: Drug delivery to the pulmonary system has been achieved by encapsulation of the drug to be delivered in microparticles having a size range between 0.5 and ten microns, preferably in the range of two to five microns, formed of a material releasing drug at a pH of greater than 6.4. In a preferred embodiment, the drug delivery system is based on the formation of diketopiperazine microparticles which are stable at a pH of 6.4 or less and unstable at pH of greater than 6.4, or which are stable at both acidic and basic pH, but which are unstable at pH between about 6.4 and 8. Other types of materials can also be used, including biodegradable natural and synthetic polymers, such as proteins, polymers of mixed amino acids (proteinoids), alginate, and poly(hydroxy acids). In another embodiment, the microparticles have been modified to effect targeting to specific cell types and to effect release only after reaching the targeted cells.

Abstract: An exogenous stimulus is applied to tissues or cells which are at risk in a subsequent surgical procedure or other intervention which induces a response by the cells that minimizes reaction to the subsequent procedure. Stimuli can be chemical, physiological or physical. Examples include those stimuli known to induce expression of stress response proteins or heat shock proteins, especially heat shock protein 70 (hsp 70) and hsp 90, for example, exposure to heat or dilute hydrogen peroxide, or direct administration of exogenous heat shock proteins, or those stimuli which act to inhibit or reduce heat shock protein expression, for example, treatment with flavonoids.

Abstract: A method for determining the probability of an episode of prolonged apnea in an infant less than or equal to 6 months of age has been developed. The method includes conducting a physiological feeding study wherein the subject is bottle fed while monitoring respiration, heart rate, O.sub.2 saturation, and sucking pressure, conducting a sleep study wherein the subject is allowed to sleep naturally while monitoring respiration, heart rate, O.sub.2 saturation, and brain activity, and analyzing the data to make a prediction of the probability of a prolonged episode of sleep apnea. The method can further include correlating the results of the physiological feeding study and the sleep study with age, epidemiologic characteristics, and birth age. In the preferred embodiment, the sleep study is conducted in an environmentally controlled test chamber at 90.degree..+-.2.degree. F., and both respiratory effort and air flow are measured.

Abstract: Hydrogels of polymerized and crosslinked macromers comprising hydrophilic oligomers having biodegradable monomeric or oligomeric extensions, which biodegradable extensions are terminated on free ends with end cap monomers or oligomers capable of polymerization and cross linking are described. The hydrophilic core itself may be degradable, thus combining the core and extension functions. Macromers are polymerized using free radical initiators under the influence of long wavelength ultraviolet light, visible light excitation or thermal energy. Biodegradation occurs at the linkages within the extension oligomers and results in fragments which are non-toxic and easily removed from the body. Preferred applications for the hydrogels include prevention of adhesion formation after surgical procedures, controlled release of drugs and other bioactive species, temporary protection or separation of tissue surfaces, adhering of sealing tissues together, and preventing the attachment of cells to tissue surfaces.