sepharose and Acute-Disease

Methylprednisolone (MP) has been shown to reduce acute inflammation resulting from a secondary damage cascade initiated by the primary physical injury to the spinal cord. The current clinical practice for delivering systemic MP is inefficient, and high doses are required, resulting in adverse, undesired, dose-related side effects in patients. Here, we report a novel, minimally invasive, localized drug delivery system for delivering MP to the contused adult rat spinal cord that potentially side-steps the deleterious consequences of systemic cortico-steroid therapy. MP was encapsulated in biodegradable PLGA based nanoparticles (NP), and these nanoparticles were embedded in an agarose hydrogel for localization to the site of contusion injury. To visualize and quantify its spatial distribution within the injured spinal cord, MP was conjugated to Texas-red cadaverine prior to encapsulation in nanoparticles. When delivered via the hydrogel-nanoparticle system, MP entered the injured spinal cord and diffused up to 1.5mm deep and up to 3mm laterally into the injured spinal cord within 2 days. Furthermore, topically delivered MP significantly decreased early inflammation inside the contusion injured spinal cord as evidenced by a significant reduction in the number of ED-1(+) macrophages/activated microglia. This decreased early inflammation was accompanied by a significantly diminished expression of pro-inflammatory proteins including Calpain and iNOS. Additionally, topically delivered MP significantly reduced lesion volume 7 days after contusion injury. The minimally invasive MP delivery system reported in this study has the potential to enhance the effectiveness of MP therapy after contusion injury to the spinal cord and avoid the side effects arising from high dose cortico-steroid therapy.

Topics: Acute Disease; Animals; Anti-Inflammatory Agents; Delayed-Action Preparations; Drug Carriers; Male; Methylprednisolone; Rats; Rats, Sprague-Dawley; Sepharose; Spinal Cord Injuries; Tissue Distribution; Treatment Outcome

Hyperacute organ xenograft rejection can be prevented by removing anti-pig antibodies by extracorporeal absorption prior to transplantation. A novel recombinant absorber of anti-pig antibodies was developed by fusing the cDNA encoding the extracellular part of a mucin-type protein, P-selectin glycoprotein ligand-1, with an antibody Fc fragment cDNA, which upon coexpression with the porcine alpha1,3 galactosyltransferase carried the xenogeneic epitope, Galalpha1,3Gal (Liu J., Qian Y., Holgersson J., Transplantation 1997, 63, 1673-1682). The biochemical characterization of the mucin/Ig and its absorption efficacy compared with that of porcine thyroglobulin and Galalpha1,3Gal-conjugated beads are reported. The carbohydrate portion of the mucin/Ig constituted 43% of its molecular weight and the majority of the Galalpha1,3Gal epitopes were O-linked as assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blotting following N-glycosidase F digestion. Gas chromatography-mass spectrometry of reduced and acetylated saccharides released by alpha-galactosidase treatment revealed that the fusion protein carried approximately 140 mol of terminal, alpha-linked galactose per mole protein. Based on the reduction in pig aortic endothelial cell cytotoxicity, Galalpha1,3Gal-substituted mucin/Igs on agarose beads were, on a carbohydrate molar basis, shown to be approximately 20 times more efficient than agarose-conjugated pig thyroglobulin, and approximately 5000 and 30,000 times more efficient than Galalpha1,3Gal-substituted agarose and macroporous glass beads, respectively. Structural features of the mucin backbone and its carbohydrate core saccharide chains determine the structural context, spatial orientation and spacing of Galalpha1,3Gal epitopes and are likely to explain the superior absorption efficacy of the recombinant mucin-type chimera.

Topics: Absorption; Acute Disease; Animals; Antibodies, Heterophile; Antigens, Heterophile; COS Cells; Disaccharides; Epitopes; Galactose; Glass; Graft Rejection; Immunoglobulin Fc Fragments; Membrane Glycoproteins; Mucins; Oligosaccharides; Recombinant Proteins; Sepharose; Swine; Thyroglobulin; Transplantation, Heterologous

The method for the preparation of a specific proteinase adsorbent (acid-resistant human urinary proteinase inhibitor--UPI--immobilized on sepharose) has been developed. Proteinase adsorption using this adsorbent has proved highly effective in the treatment of acute pancreatitis with plasmasorption. High adsorption capacity of UPI-sepharose from plasma samples of patients with acute nonspecific aortoarteritis has been demonstrated in vitro. Determination of UPI activity in the human urine is an informative test for early tissue damage, as compared to plasma middle molecules assay. The former test may be used as a diagnostic technique in patients with parenchymal kidney injury and increased arterial pressure.

Topics: Acute Disease; Adult; Animals; Aortitis; Arteritis; Dogs; Drug Evaluation; Drug Evaluation, Preclinical; Drug Stability; Female; Hemoperfusion; Humans; Pancreas; Pancreatitis; Peptide Hydrolases; Protease Inhibitors; Sepharose; Takayasu Arteritis

A newly developed peroxidase-linked immunoassay is described which is sensitive enough to quantify herpes simplex antibodies in serum and cerebrospinal fluid diluted to an IgG level of 1 mg/dl. Thus, a comparison of photometric signals allows the direct detection of specific antibodies which have been secreted by activated tissue B lymphocytes into the CSF compartment during the humoral immune phase of herpes simplex encephalitis. The technique utilizes urea-Triton-dissolved virus antigens covalently bound to Sepharose 4B pearls. A highly specific sandwich antibody was purified by immune absorption column chromatography and labelled in its protected state. In the majority of cases the antibody level increased around the 10th day, to reach its maximum a few days after. In some cases however the serum levels gradually rose over a period of several weeks. The antibody levels in the CSF increase uniformly at the same time, irrespective of the general immune response and soared up to higher than serum levels within a few days. Local antibody production may persist for years so that late diagnosis of herpes encephalitis becomes possible with a single side by side test of serum and CSF from the patient.

Topics: Acute Disease; Animals; Antibodies, Viral; Brain; Chronic Disease; Complement Fixation Tests; Encephalitis, Arbovirus; Enzyme-Linked Immunosorbent Assay; Herpes Simplex; Humans; Immunoglobulin G; Meningoencephalitis; Multiple Sclerosis; Rabbits; Sepharose

Chemotaxis, chemokinesis, and spontaneous locomotion of polymorphonuclear leukocytes (PMNs) of yersinia arthritis (YA) patients and healthy subjects with or without HLA-B27 were studied by agarose assay and membrane filter technique. HLA-B27 positive healthy control subjects and yersinia patients showed significantly higher response to chemotactic stimulus than HLA-B27 negative controls. Consequently, the high response was associated with HLA-B27 irrespective of YA. The high responder PMNs may contribute to the more severe inflammatory symptoms in YA patients with HLA-B27.

Topics: Acute Disease; Adolescent; Adult; Arthritis, Infectious; Cell Movement; Chemotaxis, Leukocyte; Female; HLA Antigens; Humans; Kinetics; Male; Middle Aged; Neutrophils; Sepharose; Time Factors; Yersinia Infections; Zymosan