Isolation of thyroid cells obtained by fine-needle aspiration biopsy.

Usually thyroid cells isolated from tissue obtained by surgery or thyroid cell lines are used to investigate the pathogenesis of autoimmune thyroid diseases. Isolation and cultivation of thyrocytes from fine-needle aspiration biopsy (FNAB) has not yet been published. The aim of this study was to isolate and cultivate thyrocytes from samples of FNAB. FNAB samples were obtained from nine adults and nine children with Hashimoto's thyroiditis (HT). The aspiration material was filtered resulting in small samples of tissue on the surface of the filter membrane. These tissue fragments were digested by collagenase I and dispase II. The yielding cells were cultivated for 3 weeks in Ham's F12 Kaighn's Modification medium in presence of 1 mU/mL bovine thyrotropin (TSH), 10 microg/mL human insulin, 6 microg/mL transferrin, and 10(-8) M hydrocortisone. Finally, isolated thyroid cells were characterized by determination of gene expression of thyrotropin receptor (TSHR), thyroperoxidase (TPO), and thyroglobulin (Tg) using a nested reverse transcriptase-polymerase chain reaction (RT-PCR). Thyroid cells obtained by FNAB can be maintained over a time period of approximately 3 weeks. Depending on the sample size a final number of 1000-14,000 cells was gained per FNAB. In addition, all cells isolated by the described method expressed TPO mRNA. TSHR mRNA was found in 4 samples, whereas 15 samples were Tg mRNA-positive. There were no differences with respect to the expression TSHR and TPO mRNA between samples from adults and children. The isolation and cultivation of thyroid cells obtained by FNAB has been established. In contrast to surgical specimen, this technique provides an easy access to thyrocytes derived from individual patients allowing repeated sampling to investigate the time progression of the chronic disease or the effect of treatment over time.

Effect of 1-year treatment with nitrendipine versus enalapril on urinary albumin and alpha 1-microglobulin excretion in microalbuminuric patients with type 1 diabetes mellitus. A randomized, single-blind comparative study.

In order to compare the long-term effects of nitrendipine (CAS 39562-70-4) and enalapril (CAS 75847-73-3) on variables of glomerular and tubular function in type 1 diabetes mellitus, a single-blind, randomised comparative 1-year study was carried out in microalbuminuric patients (6 women, 14 men, age, 30-58 years, duration of diabetes, 3-41 years, HbA1 sigma 5.5-10%). 10 patients were treated with 20 mg/d nitrendipine, 10 patients were treated with 10 mg/d enalapril. On the average, urinary albumin excretion was decreased by 38 +/- 4% by nitrendipine (p < 0.01 vs. before treatment) and by 21 +/- 8% by enalapril (p < 0.05 vs. before treatment). The excretion of alpha 1-microglobulin decreased by 35 +/- 10% and by 39 +/- 9%, respectively (p < 0.05 vs. before treatment). Creatine clearance rose by 20 +/- 30% during nitrendipine treatment (p < 0.05 vs. before treatment) but was unchanged during enalapril treatment. Total kidney volume decreased by 23 +/- 4% (p < 0.01) and by 14 +/- 6% (p < 0.05), respectively. Blood pressure fell by 8 +/- 1% systolic and by 13 +/- 1% (diastolic) in nitrendipine-treated patients (both p < 0.01) and by 10 +/- 1% and 13 +/- 1% in enalapril-treated patients (both p < 0.01). Thus, nitrendipine long-term treatment of microalbuminuric type 1 diabetic appeared to be as effective as the treatment with enalapril in preventing or postponing the progression of diabetic nephropathy.