The colour reaction was stopped after 30 min and optical density

The colour reaction was stopped after 30 min and optical density was measured at 450 nm using an MRX Revelation plate reader from Dynex Technologies (Chantilly,

VA, USA). C-peptide was measured at (NLMDRL) 6 min after stimulation with 1 mg glucagon administered intravenously, as described previously [32]. All results for T cell and C-peptide are summarized as the mean, and measures of variability are reported as standard error (s.e.). Linear regression analysis was used to determine the best-fitted line, and an analysis of covariance was used to compare slopes between groups over the entire study. Metformin order Two-tailed Mann–Whitney U-tests were used to compare results at individual time-points between the treatment

groups. Two-tailed Wilcoxon matched-pairs signed-rank tests were used to compare results between individual time-points within the treatment groups. Demographic data, islet autoantibody and T cell responses to tetanus toxoid from patients treated with rosiglitazone and glyburide are shown in Table 1. No significant differences were observed in age, sex, race, body mass index (BMI), islet autoantibodies, tetanus responses or time since diagnosis between treatment groups at baseline or 36 months (Table 1). Islet-specific T cell responses in both patient groups increased during the first 12 months, becoming selleck increased significantly (P < 0·05) compared to baseline D-malate dehydrogenase at 9 months of treatment for both patient groups (Fig. 1). However, beginning at 15 months, T cell responses to islet proteins in the rosiglitazone-treated patients became suppressed significantly (P < 0·03). In fact, the T cell responses

to islet proteins in the rosiglitazone-treated patients became negative at 15 months (fewer than four blot sections) and remained negative throughout follow-up (Fig. 1). In contrast, the T cell responses to islet proteins in the glyburide patients remained positive throughout the study (Fig. 1). Mean stimulated C-peptide responses for both glyburide- and rosiglitazone-treated patients are shown in Fig. 2. During the first 12 months of follow-up, at the time T cell proliferation increased, the C-peptide in the glyburide-treated patients remained stable, whereas the C-peptide responses in the rosiglitazone-treated patients declined significantly (P < 0·05). However, after 12 months of follow-up, when islet-reactive T cell responses were suppressed in rosiglitazone-treated patients (Fig. 1), the C-peptide responses in the rosiglitazone-treated patients improved. In contrast, the C-peptide in the glyburide patients was observed to continue to decline throughout the study, reaching significance (P < 0·05) from baseline at 36 months (Fig. 2). Comparison of the glucagon-stimulated C-peptide responses for the rosiglitazone- and glyburide-treated patients demonstrated significant differences (P < 0·05) beginning at 27 months (Fig. 2).

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