Immunohistochemical localization of thyroid hormone in rat thyroid gland.

Direct and indirect immunofluorescence techniques were used to localize the thyroid hormones triidothyronine (T3) and thyroxine (T4) in adult rat thyroid gland. Optimum dilutions of the antisera were established and four tissue fixatives were investigated for usefulness in this technique. Use of antibodies specific for either T3 or T4 resulted in brilliant fluorescence in the colloid pools and apical cytoplasm of follicular cells. In all cases, the adjacent parathyroid gland was devoid of fluorescence. This report demonstrates that these dipeptide hormones can be localized by using immunofluorescence techniques.     

Quantitative modulation of thyroid iodide peroxidase by thyroid stimulating hormone

The activity of rat thyroid iodide peroxidase fell to 8% of the normal value 48 hours after hypophysectomy. Rats given injections of thyroid stimulating hormone manifested an enzyme activity indistinguishable from that of the sham-operated animals. Cycloheximide prevented the thyroid stimulating hormone-induced restoration of the enzyme activity. The incorporation of ¹⁴C-leucine into the thyroid gland decreased gradually and reached two thirds of the sham-operated group by 48 hours after hypophysectomy. Thyroid stimulating hormone administration prevented this decrease, as observed for iodide peroxidase activity. Thyroidal RNA contents decreased also in hypophysectomized rats, thyroid stimulating hormone treatment prevented the reduction of RNA contents and no significant change was observed in thyroidal DNA contents. These data are consistent with the idea that protein biosynthesis is involved in thyroid stimulating hormone regulation of thyroidal iodide peroxidase and that the life span of the peroxidase is less than 48 hours.     

Inactivation of human thyroid stimulator by human thyroid extract.

To determine whether the thyroid stimulating activity of IGG of patients with Graves' disease is associated with the reaction with a putative human thyroid antigen, the inactivation of the property of IgG to stimulate cAMP generation in human thyroid slices incubated in vitro was studied by pretreating the IgG with human thyroidal particulate fraction. In the preliminary experiment, it was demonstrated that to cause cAMP generation stimulation, on incubation period of 120 min is required to allow the IgG to penetrate the tissue. When human thyroid slices were incubated with normal IgG without or with pretreatment by human thyroid particulate fraction obtained from 100 mg tissue, cAMP content in the slices was 142 +/- 25 or 138 +/- 26 f moles/mg, respectively, indicating that basal thyroidal cAMP levels were not influenced at all by normal IgG even after pretreatment with thyroid particulate fraction. When the slices were incubated with gG of Graves' disease without or with the similar pretreatment, cAMP content was 320 +/- 31 or 140 +/- 25 f moles/mg, respectively, demonstrating an almost complete inhibition of the activity of the IgG to cause cAMP generation stimulation.     

Immunohistochemistry of thyroid hormones as a functional parameter in thyroid pathology

The authors study by means of immunoperoxidase method the pattern of thyroglobulin, triiodothyronine and thyroxine distribution in 58 cases of thyroid disorders: 15 euthyroid goiters, 10 Graves' disease, 7 Hashimoto's thyroiditis, 11 folliculo-papillary carcinomas (6 primary tumors and 5 lymph node metastases), 8 follicular carcinomas, 4 anaplastic carcinomas and 3 medullary carcinomas. Thyroglobulin, triiodothyronine and thyroxine were present in most of the thyroid disorders, excepting anaplastic and medullary carcinomas. Thyroglobulin and thyroxine were localized both in the follicular epithelium and in the colloid, whereas triiodothyronine was present especially in the follicular cells. The thyroid hormones distribution in benign lesions is rather similar. In carcinomas, the pattern of thyroglobulin, triiodothyronine and thyroxine is more heterogeneous, but generally the triiodothyronine distribution is similar to that of thyroglobulin. In some carcinomas, triiodothyronine and thyroxine showed a weak or negative immunostaining. The immunoperoxidase method is a valuable tool in the study of functional disturbances in the thyroid pathology and in the diagnosis of thyroid carcinoma metastases as well. Positive thyroid hormones staining clearly indicates the thyroid origin of metastases.     

Stimulation of iodine organification in porcine thyroid cells by thyroid stimulators

Several Graves' sera were simultaneously assessed in a bioassay based on the ability of porcine thyroid cells to organify 125I and in a radioreceptor assay for TSH receptor binding activity. Both assay systems were sensitive to 1 mcU/ml (final concentration) of unlabelled bovine TSH. Six Graves' sera were studied in detail over a wide (0-1.0 mcl sera) dose response range in repeat determinations. Two sera exhibited parallel binding and stimulating. However, two sera revealed significant inhibition of 125I-TSH binding prior to the demonstration of stimulation and the other two sera showed stimulatory capabilities before significant binding was evident. IgG was prepared from one serum by ammonium sulphate precipitation and chromatography on Sepharose 6B and then subjected to preparative isoelectric focusing. The isoelectric distribution of the two activities were found to be identical with major peaks of activity at pl=9.5 and pl=8.5. In summary: 1) each Graves' sera exhibits different dose-response curves with respect to binding and stimulation, 2) at certain concentrations of sera, only binding or stimulation were evident, 3) neither assay was consistently more sensitive for the presence of Graves' immunoglobulins, 4) for one Graves' sera, binding and stimulation could not be separated by isoelectric focusing. These studies would suggest each Graves' immunoglobulin has inherently different characteristics in its interaction with the TSH receptor.     

Measurement of thyroid hormone in the rat thyroid gland by radioimmunoassay

The present paper describes (i) a hydrolysis technique with Pronase and leucine aminopeptidase using one rat thyroid gland, resulting in maximum release of thyroid hormones and minimum deiodination, and (ii) a simple and rapid procedure for thyroid hormone radioimmunoassays in thyroid hydrolysates using commercial kits intended for serum thyroid hormone determinations. The procedure is used to determine T4, T3, and rT3 concentrations and hormonal molar ratios in a thyroid gland from a male Wistar rat. The reliability of the method is also studied.     

Prenatal programming of adult thyroid function by alcohol and thyroid hormones

Increasing evidence associates environmental challenges early in life with permanent alterations of physiological functions in adulthood. These changes in fetal environment can trigger physiological adaptations by the fetus, called fetal programming, which may be beneficial before birth but permanently influence the physiology of the organism. In this study, we investigated the potential connection between alcohol-induced decreased maternal thyroid function and the hypothalamic-pituitary-thyroid (HPT) function of adult rat offspring. Plasma 3,5,3'-triiodothyronine (T(3)), thyroxine (T(4)), and thyroid-stimulating hormone (TSH) levels were decreased in alcohol-consuming (E) dams on gestational day 21 compared with ad libitum- (C) and pair-fed (PF) controls. No significant differences were found in HPT function in young offspring (3 wk of age) between diet groups. However, adult fetal alcohol-exposed (FAE) offspring had significantly decreased levels of T(3) along with elevated TSH compared with control offspring. T(4) administration to the mother did not normalize the hypothyroid state of the adult FAE offspring. Interestingly, administration of T(4) to control pregnant dams decreased plasma T(3) of the adult female offspring only, whereas T(4) together with maternal alcohol consumption or pair-feeding led to decreased TSH and T(4) in the adult female offspring. Our results suggest that ethanol consumption and T(4) administration alter maternal HPT function, leading to prenatally programmed permanent alterations in the thyroid function of the adult offspring.     

Timing of repeat thyroid fine-needle aspiration in the management of thyroid nodules

Objective: The Bethesda System for reporting thyroid fine-needle aspirations (FNAs) recommends repeat aspirations be done at least 3 months following the initial aspiration to prevent false-positive misinterpretations due to reactive/reparative changes. Because limited data exists, we reviewed our own data to ascertain the recommendation. Study Design: We retrospectively reviewed thyroid FNAs accessioned in the Cytology Laboratory at Beth Israel Deaconess Medical Center in Boston, Mass., USA, from January 2006 to December 2008. We identified patients with repeat aspirates of a single lesion and those with thyroidectomy specimens. Cytologic and surgical diagnoses along with time between FNAs were recorded. Results: No statistically significant difference was detected in the distribution among nondiagnostic, suboptimal, and adequate categories with respect to the timing of repeat FNA (p = 0.25). Forty-six percent of the lesions that were nondiagnostic/suboptimal initially yielded the same results on repeat. Twenty-eight percent of initially adequate specimens were nondiagnostic/suboptimal on repeat (p = 0.04). Nine percent of those with an initial atypical diagnosis had the same diagnosis on repeat, while 2.6% of those who did not have an initial atypical diagnosis had an atypical diagnosis on repeat (p = 0.29). False positives between early (16%) and late (12%) repeats were not statistically significant (p = 0.74). Conclusion: The diagnostic yield and accuracy of repeat FNA is independent of the time interval between procedures but may be related to the original FNA diagnosis.