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Mechanisms of Shiga toxin-mediated signaling and toxicity

AbstractShiga toxin (Stx) is the main virulence factor of enterohemorrhagic Escherichia coli (EHEC). EHEC strains cause gastrointestinal infection and release Stx that can gain access to the circulation. Patients may develop hemolytic uremic syndrome with extensive kidney damage. In the first paper we investigated if blood cell-derived microvesicles released during EHEC infection contain Stx. Toxi

Framework of Last Mile Logistics Research: A Systematic Review of the Literature

Coincident with the rapid growth of omni-channel retailing, growing urbanization, changing consumer behavior, and increasing focus on sustainability, academic interest in the area of last mile logistics has significantly increased. The growth in academic publications has been tremendous, with three out of four articles appearing within the past five years. The influx of research spans multiple dis

Phenotypic antibody discovery and mining of complex antibody libraries

Therapeutic antibodies are the fastest growing class of drugs but suffer from a crowded target-space where many antibodies, against a few targets, are developed in parallel. The aim of this thesis is to find methods to enable discovery of novel antibody-target combinations and thereby increase the target-space. A strategy to achieve this is to use phage display antibody libraries and selection on

High diagnostic accuracy for idiopathic Addison’s disease with a sensitive radiobinding assay for autoantibodies against recombinant human 21-hydroxylase

Autoantibodies against 21-hydroxylase (P450c21) are common in idiopathic autoimmune Addison’s disease. In the present work, we have developed a sensitive radiobinding assay using in vitro translated recombinant human 35S-P450c21. Levels of P450c21 antibodies (P450c21-Ab) were expressed as a relative index (P450c21 index) using a P450c21-Ab positive Addisonian serum and two antibody-negative health

Islet cell antibodies, but not glutamic acid decarboxylase antibodies, are decreased by plasmapheresis in patients with newly diagnosed insulin-dependent diabetes mellitus

The effects of plasmapheresis on islet autoantibody levels, C-peptide (β-cell function), and hemoglobin-A1c (HbA1c, metabolic control) were tested in a prospective blinded study of 18 newly diagnosed insulin-dependent diabetes mellitus (IDDM) patients randomly assigned to receive plasmapheresis (P), carried out as double filtration, or sham (S) treatment at diagnosis and 3 months thereafter. At di

Serum exchange and use of dilutions have improved precision of measurement of islet cell antibodies

In an attempt to improve the diagnostic value of measuring antibodies to islet cell cytoplasmic antigen, coded sera were distributed to 38 laboratories and results were returned for analysis. Comparison between laboratories revealed that results for some individual sera differed by up nine doubling dilutions and even within laboratories duplicate samples could differ by six doubling dilutions. By

[24] Preparation of Stable Radioiodinated Polypeptide Hormones and Proteins Using Polyacrylamide Gel Electrophoresis

This chapter discusses the preparation of stable radioiodinated polypeptide hormones and proteins using polyacrylamide gel electrophoresis. Radioactively labeled polypeptide hormones and proteins are widely used as tracers in radioimmunoassays and receptor studies. The peptide or protein is most easily labeled using iodination with 125I or 131I. The radioactive iodine is substituted in the tyrosin

Plasmapheresis in the initial treatment of insulin-dependent diabetes mellitus in children

Several factors indicate that autoimmune mechanisms may play a part in the aetiology of insulin-dependent diabetes mellitus. At the onset of the disease in 10 children (aged 11-16 years) plasmapheresis was performed four times over one to two weeks. Seventeen age-matched children with the same clinical features served as controls. The C-peptide concentrations at onset were the same in the two grou

Expression of major histocompatibility antigens on pancreatic islet cells

Insulin-independent diabetes mellitus is often accompanied by manifestations of autoimmunity and is frequently associated with certain HLA haplotypes, predominantly DR3 and DR4. Because the major histocompatibility antigens are important determinants of the immune response in various tissues, we have investigated their expression on the pancreatic islet cells. Human, mouse, or rat islets of Langer

Stable lodinated polypeptide hormones prepared by polyacrylamide gel electrophoresis

Iodination of several insulin and proinsulin preparations, human growth hormone and bovine pancreatic polypeptide was performed using H2O2 and lactoperoxidase or chloramine T. The iodination mixtures were fractionated by polyacrylamide gel electrophoresis at pH 9.15 in long gel rods followed by simple elution of the iodinated products from thin gel slices. With this method 125I tracers with long s

Trypan Blue as a marker of plasma membrane permeability in alloxan-treated mouse islet cells

Suspensions of pancreatic islet cells from noninbred ob/ob-mice were incubated with Trypan Blue. Microscope photometry showed that apparently viable cells excluded the dye completely, whereas the nuclei of nonviable cells accumulated Trypan Blue by a saturable process. The nucleus-to-medium dye gradient was more then 30∶1 in media containing 0.1% or less Trypan Blue. The apparent affinity constant