Characterization of triton-solubilized TSH receptors from human thyroid plasma membranes

The TSH receptor from human thyroid plasma membranes has been solubilized in 10 mM Tris/HCl, 50 mM NaCl, pH 7.4 containing 0.5% triton X-100. Binding of [125I]TSH to the soluble receptor showed rapid and reversible kinetics and reached a maximum within 30 min at 37 degrees C, by 1 h at 25 degrees C and by 24 h 4 degrees C. Optimal pH was 7.4. The soluble receptor retained specificity with cross-reactivity only to crude hCG (0.03%). Scatchard plots were curvilinear indicating the presence of at least two binding sites. The high affinity site showed an affinity content of 1.1 X 10(9) M-1 with binding capacity of 1.3 X 10(-10) M/mg protein. TSH-binding inhibitor immunoglobulins from patients with Graves' disease inhibited [125I]TSH binding to the soluble receptor in a dose-dependent manner. NaCl inhibited the TSh binding and this was ascribed to the decrease in the receptor capacity. Trypsin, neuraminidase and phospholipase C treatment of the solubilized receptor had no effect on TSH binding. The apparent molecular weight of the receptor, determined by gel filtration of Sepharose 6B, was approximately 300 000.     

Altered balance of immunoregulatory T lymphocyte subsets in autoimmune thyroid diseases

Three monoclonal antibodies recognizing cell surface antigens of total peripheral (OKT3), helper/inducer (OKT4) and suppressor/cytotoxic (OKT8) T lymphocytes were used by an indirect immunofluorescence technique to enumerate peripheral T lymphocytes in 25 patients with Graves' disease (including 4 euthyroid Graves' patients), 16 patients with Hashimoto's thyroiditis and 22 normal controls. Total lymphocyte count and percentages of overall T and helper/inducer T cells among peripheral lymphocytes in these conditions showed no significant difference from those of the controls. Percentage of suppressor/cytotoxic T cells, however, was decreased in Graves' disease patients with or without hyperthyroidism. The ratio of helper/inducer T cells to suppressor/cytotoxic T cells was increased in Graves' disease population and slightly increased in hypothyroid Hashimoto's thyroiditis patients. The ratio correlated with the mitogenic response of peripheral mononuclear cells to phytohaemagglutinin, but not with the serum levels of thyroid hormones nor with the titres of thyroid autoantibodies. These findings are in accordance with the results of previous functional studies and indicate possible defects in suppressor T lymphocytes in autoimmune thyroid disease.     

<Superscript>18</Superscript>F-FDG PET in the detection of extrahepatic metastases from hepatocellular carcinoma

Background Positron emission tomography (PET) with ¹⁸F-fluoro-2-deoxy-d-glucose (¹⁸F-FDG) is useful in detecting distant metastases from a variety of malignancies. However, its efficiency in detecting distant metastases from hepatocellular carcinoma (HCC) has not been investigated. The aim of this study was to evaluate the usefulness of ¹⁸F-FDG PET for the detection of extrahepatic metastases from HCC.Methods Nineteen patients suspected of having extrahepatic HCC underwent ¹⁸F-FDG PET. Fourteen patients (group A) had extrahepatic lesions, which were detected by conventional studies. In five patients (group B), conventional imaging showed no extra- or intrahepatic lesions, but the tumor marker levels were elevated. The PET results were compared with those obtained by histopathology or by clinical follow-up.Results The detection rate of ¹⁸F-FDG PET was 83% (24 of 29 metastases) for extrahepatic metastases larger than 1cm in greatest diameter and 13% (1 of 8 metastases) for lesions less than or equal to 1cm. PET revealed two bone metastases not depicted by bone scan, and detected the nodal metastasis and intestinal metastases inconclusive on computed tomography. Extrahepatic lesions were resected in 5 patients of group A on the basis of PET findings. In all patients of group B, PET results were true negative for extrahepatic metastases, but HCCs were detected in the liver within 4 months in 4 patients. These were no false-positive lesions in either group.Conclusions This preliminary study suggested that ¹⁸F-FDG PET could provide additional information and contribute to the management of HCC patients suspected of having extrahepatic metastases.     

Value of stress thallium-201 emission tomography for predicting improvement after coronary bypass grafting and assessing graft patency

To examine the value of stress and redistribution thallium-201 single photon emission computed tomography (SPECT) for predicting improvement after coronary artery bypass grafting (CABG) and for assessing graft patency, 17 patients underwent SPECT before and after CABG. Preoperatively, 16 patients (94%) showed reversible ischemia with redistribution, and defect score decreased from 16.4 +/- 7.6 to 8.3 +/- 5.8 (p less than 0.001) on 2 hour delayed images. Stress electrocardiography was positive in 15 cases (88%). The stress defect score was reduced postoperatively (7.8 +/- 6.3) as compared to the preoperative score (16.4 +/- 7.6) (p less than 0.001). Postoperatively, 7 cases (41%) showed reversible ischemia and the defect score decreased from 11.3 +/- 6.5 to 6.1 +/- 4.7 (p less than 0.05), while 4 cases (23.5%) showed positive stress electrocardiograms (ECG). Resting left ventricular (LV) ejection fraction (EF) improved in 6 cases (48.3 +/- 5.4% preoperatively to 56.8 +/- 7.4% postoperatively, p less than 0.05) and showed a reduction in 11 (58.2 +/- 10.5% to 47.5 +/- 9.4%, p less than 0.01). Grafts were occluded in 6 cases, and redistribution was evident on the delayed thallium images in all cases (100%), while stress ECG was positive in only 3 (50%). Thus, stress and redistribution SPECT is valuable for identifying patients who will improve after surgery and assessing graft patency more accurately than stress ECG. Resting LVEF was less useful for assessing improvement in LV function.     

Detection and properties of TSH-binding inhibitor immunoglobulins in patients with Graves' disease and Hashimoto's thyroiditis.

TSH-binding inhibitor immunoglobulins (TBII) have been detected in patients with Graves' disease and Hashimoto's thyroiditis by using the radioreceptor assay of TSH. In untreated Graves' patients, TBII levels correlated well with thyroidal 99mTc uptake at 30 min and the grade of epithelial hyperplasia of thyroid follicles. There were many Graves' patients whose sera contained high TBII levels but no detectable bioassayable thyroid-stimulating activity (LATS), and in these patients, close correlation was observed between serum levels of TBII and bioassayable LATS-protector activity. TBII were detectable in 2 (10%) of 20 patients with Hashimoto's thyroiditis, both of whom were clinically hypothyroid. The serum or IgG fraction from one of them, however, did not contain any significant LATS, LATS-protector, or human thyroid adenylate cyclase-stimulating activity and caused inhibition of adenylate cyclase stimulation by TSH. In that patient, TBII may be acting to block TSH binding to TSH receptors, thus causing TSH unresponsiveness and hypothyroidism.