These results suggest a substantial contribution of Fas/ FasL interactions to CD4؉ , but not CD8 ؉ , T-cell-mediated destruction of pancreatic ␤-cells. When anti-FasL antibody was administered to NOD mice between 5 and 15 weeks of age, the onset of diabetes was slightly delayed but the incidence was not decreased. However, administration of anti-FasL antibody at 2-4 weeks of age completely prevented insulitis and diabetes. These results suggest that Fas/FasL interactions contribute to CD4 ؉ T-cell-mediated ␤-cell destruction and play an essential role in the initiation of autoimmune NOD diabetes.

Fluoride has been used to prevent caries in the dentition, but the possible underlying mechanisms of cytotoxicity induction by this compound are still unclear. Since fluoride is known as an inhibitor of glycolytic enzymes, we investigated the possible connection between NaF-induced apoptosis and glycolysis in human promyelocytic leukemia HL-60 cells. NaF-induced apoptotic cell death is characterized by caspase activation, internucleosomal DNA fragmentation, loss of mitochondrial membrane potential, and production of apoptotic bodies. Higher activation of caspases-3 and -9, as compared with that of caspase-8, suggested the involvement of an extrinsic pathway. Utilization of glucose was nearly halted by NaF, whereas that of glutamine was rather enhanced. NaF enhanced the expression of Bad protein, but not that of Bcl-2 and Bax proteins, and reduced HIF-1alpha mRNA expression. Analysis of these data suggests a possible link between glycolysis and apoptosis.

OBJECTIVE -To investigate the prevalence of ␤-cell autoantigen-reactive peripheral T-cells in type 1 diabetes, we developed an immunoglobulin-free enzyme-linked immunospot (ELIS-POT) assay and assessed its usefulness for diagnosing this disease. RESEARCH DESIGN AND METHODS-Cellular immune responses to ␤-cell autoantigens were studied both by immunoglobulin-free proliferation assays and ELISPOT assays in 33 patients with type 1 diabetes and 15 patients with type 2 diabetes, compared with 23 healthy control subjects. Autoantibodies against GAD65 and IA-2 were measured by radioimmunoassay.RESULTS -Significant proliferative responses to GAD65 were observed in 10 of 31 (32.3%) type 1 diabetic patients (P Ͻ 0.05), whereas GAD65-reactive ␥-interferon (IFN-␥)-secreting cells were detected in 22 of 33 patients (66.7%) by ELISPOT assay (P Ͻ 0.001). Of patients negative for both GAD65 and IA-2, five of six (83.3%) showed IFN-␥ positivity in ELISPOT and two of five (40.0%) showed significant proliferation against GAD65.CONCLUSIONS -Using a newly developed ELISPOT assay, GAD-reactive T-helper 1 cells in PBMC of type 1 diabetic patients could be identified at a higher frequency than by the proliferation assay. Therefore, the immunoglobulin-free ELISPOT assay is an excellent tool for detecting T-cell reactivity to autoantigens with greater specificity and, in combination with ␤-cell autoantibody determination, will improve the diagnosis of type 1 diabetes. Diabetes Care 25:1390 -1397, 2002S everal studies of type 1 diabetes have suggested that the destruction of pancreatic ␤-cells is caused by inflammatory cellular immune responses mediated by T-helper 1 (Th1) cells, which secrete ␥-interferon (IFN-␥) (1-3). In diabetic animal models, a correlation between disease onset and Th1 cytokine production has been observed, and IFN-␥ in particular plays an important role in ␤-cell destruction (4 -6). On the other hand, autoantibodies against GAD65, insulin, IA-2/ICA512, and other islet cell antigens have been used not only to diagnose but also to predict the onset of type 1 diabetes (7-9). However, differences in age at onset of diabetes as well as racial differences have been reported to influence patients' autoantibody status (10,11). In general, circulating levels of autoantibodies are considerably lower in patients with a long disease duration than in patients at the time of onset or in the prediabetic period (12). According to a Japanese multicenter study, positivity for GAD antibody was found in only 35.4% of 921 Japanese type 1 diabetic patients studied, but among those with newly diagnosed diabetes, 50.3% were GAD antibody positive (13). Japanese type 1 diabetic subjects, therefore, seemed to present with lower antibody levels than Caucasians. In addition, it has been suggested that the presence of GAD antibody can identify a subset of latent autoimmune diabetes in adults (LADA) and predict incipient insulin dependency (14,15). Indeed, the patient's autoantibody status is essential for the diagnosis and prediction of type 1 di...

Thirty percent of the total number of patients discontinued treatment because of side effects or unsatisfactory pain relief. For those continuing treatment, clinical improvements were seen in the initial 6 months in both groups. Fifty percent of the TDB and 43% of TDF groups had significant relief in 3 months, which persisted up to 6 months. Only 11% and 13% of patients, respectively, had sustained relief after 6 months. Twenty percent more patients in the TDB group benefited significantly in symptoms of depression from TDB compared with the TDF group. Interestingly, switching of patches seemed to increase acceptability by preventing adverse effects and tolerance. Confirmation of these effects should be studied in future with a multicenter study and larger sample.