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The World Health Organization international collaborative study for islet cell antibodies

Aims/hypothesis. Islet cell autoantibodies are a specific marker for Type 1 (insulin-dependent) diabetes mellitus. Standardisation of islet cell antibodies and the uniform reporting in International units is critical to research and the development of assays for islet cell autoantibodies as diagnostics. Methods. The suitability of a candidate serum to serve as the international standard for islet

Evaluation of a novel radioimmunoassay using 125I-labelled human recombinant GAD65 for the determination of glutamic acid decarboxylase (GAD65) autoantibodies

Autoantibodies to the islet cell 65-kilodalton isoform of glutamic acid decarboxylase (GAD65) are present in most patients with type 1 diabetes mellitus years before the clinical manifestation of the disease. GAD65 autoantibodies are also present in a subset of patients with type 2 diabetes who frequently become insulin dependent. In the present study, we evaluated a new, commercially available ra

Site-directed mutagenesis of K396R of the 65 kDa glutamic acid decarboxylase active site obliterates enzyme activity but not antibody binding

The role of K396 in the enzymatic catalysis and the antigenicity of the 65 kDa isoform of glutamate decarboxylase (GAD65) was analyzed using the K396R GAD65 mutant. GAD65 is a major autoantigen in Type 1 diabetes and autoantibodies directed to GAD65 are widely used markers for this disease. We found that (1) recombinant human GAD65 is fully enzymatically active; (2) the K396R mutation abolished GA

Activated human epitope-specific T cells identified by class II tetramers reside within a CD4high, proliferating subset

Antigen-specific T cells acquire a distinctive phenotype during activation, with characteristic acquisition of surface markers and patterns of gene expression. Early after antigen stimulation, CD4+ T lymphocytes increase their surface density of the CD4 marker, a trait which has been used to identify antigen-activated cells. The recent development of MHC tetramer technologies has greatly improved

Treatment outcome for IDDM patients in relation to glutamic acid decarboxylase autoantibodies and serum IgG to periodontal pathogens

Background: Patients with insulin-dependent diabetes mellitus (IDDM) have elevated risk for periodontitis (PD) relative to subjects without diabetes. Whether refractory PD in IDDM patients is related to autoimmunity as indicated by serum glutamic acid decarboxylase autoantibody GAD Ab levels or to host bacterial immunity as reflected by serum antibody titers to periodontal pathogens is unknown. Ai

Preservation of enzyme activity and antigenicity after mutagenesis of the membrane anchoring domain of GAD65

The smaller isoform of glutamic acid decarboxylase, GAD65, is an important autoantigen implicated in the pathogenesis of type 1 diabetes whereas the larger isoform, GAD67 appears to play no major role. The primary difference between the two isoforms resides in the N-terminal part of the molecule including the GAD65 membrane-anchoring domain. The aim of this study was to generate mutants of the mem

Low-dose streptozotocin induces sustained hyperglycemia in Macaca nemestrina

The potential for using macaques to create a nonhuman diabetic model was investigated. The significant objectives were to determine a) prognosis of STZ induced permanent beta cell destruction in nonhuman primates, and b) the potential to use STZ treated animals in a model of autoimmune diabetes by following adoptively transferred lymphocytes into MHC identical macaques. Beta cell impairment was ac

Bag-limits på torsk i Öresund

Fritidsfisket är en viktig källa till rekreation samtidigt som företag inom fisketurism bidrar till utvecklingen av de kustnära regionerna. Sedan januari 2017 regleras fritidsfisket på torsk i västra Östersjön av en bag-limit. Det innebär att det är tillåtet att ta hem högst fem torskar per dag (tre under lekperioden, februari-mars). I det berörda området är många fisketurismföretag så kallade tur

Low serum conditions for in vitro generation of human macrophages with macrophage colony stimulating factor

Animal serum is often used to generate human macrophages in vitro. Since fetal calf serum (FCS) may complicate antigen uptake, processing and presentation on HLA molecules, we tested the ability of M-CSF to generate macrophages at low fetal calf serum conditions. Peripheral blood monocytes from 12 individuals were cultured 1-4 days with 0-100 ng/ml macrophage colony stimulating factor (M-CSF) at e

Glucose-regulated insulin expression in diabetic rats

Retroviral vectors encoding glucose-responsive promoters driving furin expression may provide an amplified, glucose-regulated secretion of insulin. We constructed LhI*TFSN virus to encode a glucose-regulatable transforming growth factor α promoter controlling furin expression with a viral LTR promoter driving constitutive expression of furin-cleavable human proinsulin. Autologous BB rat vascular s

Gene and Cell-Replacement Therapy in the Treatment of Type 1 Diabetes : How High Must the Standards Be Set?

Recent advances in molecular and cell biology may allow for the development of novel strategies for the treatment and cure of type 1 diabetes. In particular, it is now possible to envisage restoration of insulin secretion by gene or cell-replacement therapy. The β-cell is, however, remarkably sophisticated, and many of the features of this highly differentiated secretory cell will have to be faith

Autoimmune type 1 diabetes : Resolved and unresolved issues

Based on the presence of autoantibodies and a strong HLA linkage, type 1 diabetes is now classified as a chronic autoimmune disease. Many issues, however, remain unresolved. Although autoantibodies to GAD65, IA-2, and insulin are clearly markers for this disease, it is not known whether they contribute to pathogenesis or are simply the response to an existing underlying destructive process. Based

Prediction of diabetes with body mass index, oral glucose tolerance test and islet cell autoantibodies in a regional population

Objective. Our aim was to test the hypothesis that a combination of markers for Type 1 diabetes (glutamate decarboxylase and IA-2 autoantibodies) and for Type 2 diabetes [oral glucose tolerance test (OGTT) and body mass index (BMI)], would predict clinical diabetes in a regional population. Design. A population-based follow-up cohort study. Setting. Participants visited the primary health care cen

A novel monoclonal antibody specific for the N-terminal end of GAD65

We describe a novel monoclonal antibody raised towards the N-terminus of the 65 kDa isoform of glutamate decarboxylase (GAD65). This N-GAD65 mAb is highly specific for GAD65 and can be used in a wide range of applications such as Western blot analysis, immunoprecipitation and immunocytochemistry. Full-length cDNAs coding for N-GAD65 mAb heavy and light chains were cloned and characterized by nucle

Induction of islet allotolerance in nonhuman primates

Recent clinical trials have pioneered the successful use of a nonsteroidal immunosuppressive regimen and established a basis for application in a routine clinical setting. In this study, a single islet transplant was not sufficient to regulate blood glucose levels, and a second transplant became necessary. A similar observation was made in our macaque islet transplant study, where animals after th

Proportional–Integral Controller Design for Combustion-Timing Feedback, from n-Heptane to iso-Octane in Compression-Ignition Engines

The problem of designing robust and noise-insensitive PI controllers for pressure-sensor based combustion-timing control was studied through simulation. Different primary reference fuels (PRF) and operating conditions were studied. The simulations were done using a physics-based, control-oriented model with an empirical ignition-delay correlation. It was found that the controllable region, in-betwThe problem of designing robust and noise-insensitive PI controllers for pressure-sensor based combustion-timing control was studied through simulation. Different primary reference fuels (PRF) and operating conditions were studied. The simulations were done using a physics-based, control-oriented model with an empirical ignition-delay correlation. It was found that the controllable region, in-betw