Human telomerase reverse transcriptase gene expression and the surgical management of suspicious thyroid tumors.

PURPOSE: Patients with a preoperative cytologic diagnosis of a suspicious thyroid nodule present a therapeutic dilemma because surgery differs for benign and malignant lesions. To address this issue, several molecular markers, including human telomerase reverse transcriptase (TERT), have been tested as markers of thyroid cancer. Because most studies select cases falling into well-defined categories to test new markers, they may overestimate their discriminatory power when applied to samples that are difficult to classify. Fine-needle aspirates (FNAs) of the thyroid with indeterminate cytology are an example of such cases. EXPERIMENTAL DESIGN: We examined whether assessing TERT mRNA by reverse transcription-PCR could have improved the surgical management in a cohort of 100 patients undergoing thyroidectomy for indeterminate FNA results. RESULTS: Ninety percent of 48 cancers were TERT positive, as were 35% of 52 benign lesions. When 10 cases with concomitant lymphocytic thyroiditis were excluded, the overall sensitivity of TERT was 91% (95% confidence interval, 80-98%) and specificity was 79% (64-90%). No clinical or tumor variable contributed to the predictive ability of TERT except for tumor size, which added only marginally. Basing the surgical approach on the TERT assay alone would have reduced lobectomies performed for malignant disease from 11 to 4 cases and reduced total thyroidectomies for benign lesions from to 15 to 9, an overall 50% reduction in suboptimal treatment. CONCLUSIONS: The overall performance of preoperative differential diagnosis for thyroid tumors with indeterminate FNA results can be substantially improved by the inclusion of molecular markers such as TERT.

Cdc25A and cdc25B expression in malignant lymphoma of the thyroid: correlation with histological subtypes and cell proliferation.

Cdc25B and cdc25A phosphatases are representative stimulators of cell cycle progression, and recent studies have also indicated their oncogenic roles. In this study, we investigated the expression of these phosphatases in malignant lymphoma of the thyroid by immunohistochemistry. These phosphatases were not expressed in follicular cells in normal follicles, but were heterogeneously or diffusely expressed in the follicles in chronic thyroiditis and malignant lymphoma. In infiltrating lymphocytes in chronic thyroiditis, they were only occasionally expressed. Of the 47 cases of lymphoma, 30 (63.8%) were classified as high group for cdc25B because it was expressed in more than 25% of lymphoma cells. Cdc25B expression level was inversely associated with MIB-1 labeling index (p=0.0008), and aberrant p53 expression (p=0.0077). Furthermore, cases of marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MZBL) were more frequently classified as high group (p=0.0318) than those of diffuse large B-cell lymphoma (DLBL). On the other hand, 22 cases (46.8%) were regarded as high group for cdc25A, but its expression level was not linked to those parameters. These findings suggest that i) cdc25B plays a role in the early phase of thyroid lymphoma possibly including the malignant transformation from chronic thyroiditis, and ii) cdc25A may contribute to the progression of lymphoma.

Caracterization of b-lymphocyte activity in thyroid glands of Graves' disease

Foram analisados por imuno-histoquímica 30 tecidos tireodianos parafinados de pacientes com doença de Graves tratados com drogas antitireoidianas. O método aplicado foi avidina-biotina-peroxidase, utilizando-se anticorpos monoclonais específicos para lonfócitos B (CD20) e macrófagos (CD68) e anticorpos policlonais para caracterizaçäo de imunoglobulinas de classes IgA,M,G cadeias leves Kappa e Lambda. Os resultados foram comparados com sete espécimes de tecido tireoidiano normal. Constatou-se uma positividade de 40 por cento para CD20 e diferenças significativas para IgA (p=0,0065) Kappa (p=0,017) e Lambda (p=0,0016). A presença de imunoglobulinas confirma o envolvimento de elementos humorais, além dos celulares, na patogênese e persistência dos mecanismos auto-imunes na doença de Graves

Cholesterol 7 alpha-hydroxylase activity in hypothyroidism and hyperthyroidism in humans.

Alterations of serum cholesterol levels are well recognized findings in hypothyroidism and hyperthyroidism. It remains unclear, whether thyroid hormones may affect serum concentrations of cholesterol through changes in the activity of cholesterol 7 alpha-hydroxylase, the rate-limiting enzyme in the catabolic conversion of cholesterol to bile acids. We determined serum concentrations of the bile acid precursor 7 alpha-hydroxy-4-cholesten-3-one, which reflects cholesterol 7 alpha-hydroxylase activity in the liver, in 19 patients with hypothyroidism and in 10 patients with hyperthyroidism before and after treatment, respectively. In patients with hypothyroidism, serum concentrations of cholesterol and LDL-cholesterol decreased by 33% (p < 0.0005) and 39% (p < 0.0005), respectively, after replacement therapy with thyroid hormones. In contrast, serum concentrations of 7 alpha-hydroxy-4-cholesten-3-one (21.7 +/- 15.8 ng/ml vs 24.5 +/- 18.1 ng/ml before treatment, n.s.) as well as serum HDL-cholesterol were unchanged during substitution therapy. In patients with hyperthyroidism, serum concentrations of cholesterol and LDL-cholesterol increased by 27% (p < 0.01) and 39% (p < 0.01) after antithyroid treatment, respectively. Again, serum concentrations of 7 alpha-hydroxy-4-cholesten-3-one did not change significantly during treatment (15.8 +/- 12.6 ng/ml vs 14.7 +/- 8.1 ng/ml before treatment, n.s.). These findings indicate that in humans, thyroid hormones influence serum lipid concentrations by other mechanisms than by affecting the activity of cholesterol 7 alpha-hydroxylase.

Significant correlation of telomerase activity in thyroid papillary carcinomas with cell differentiation, proliferation and extrathyroidal extension.

Telomerase activity was examined by telomeric repeat amplification protocol assay in thyroid disease states, including adenomas and carcinomas, and correlated with clinicopathological features. Of a total of 26 papillary carcinomas, 16 cases (61.5%) were positive, with the poorly differentiated subtype being predominant (P < 0.05). A significantly more shortened terminal restriction fragment length (P < 0.05), higher incidence of extrathyroidal extension (P < 0.001), and more elevated Ki-67 labeling indices (P < 0.002) were also found in telomerase-positive than in telomerase-negative papillary carcinomas. Of four follicular carcinomas, 3 cases (75.0%) were positive. Positive telomerase activity in follicular adenomas (9/23 cases, 39.1%) and lymphocytic thyroiditis (12/22 cases, 54.5%) appeared to be mainly caused by infiltrating lymphocytes. However, three cases of atypical adenoma with relatively increased Ki-67 labeling indices were positive, suggesting a possibility of malignant potential. The good correlations with extrathyroidal invasiveness, Ki-67 labeling indices and poor differentiation of papillary carcinomas, established by multivariate analysis, suggest that this parameter might have potential application in the estimation of tumor progression and prognosis, and in clinical management.

Urinary N-acetyl-beta-D-glucosaminidase (NAG) activity in patients with Graves' disease, subacute thyroiditis, and silent thyroiditis: a longitudinal study.

Urinary N-acetyl-beta-D-glucosaminidase (NAG) activity was measured longitudinally in 12 patients with Graves' disease, 5 patients with subacute thyroiditis, and 1 patient with silent thyroiditis, and compared with that of 36 normal controls. The patients with Graves' disease and subacute thyroiditis were treated with anti-thyroid drug (methimazole or propylthiouracil) and prednisolone, respectively. On the other hand, no treatment was given to the patient with silent thyroiditis. Since two patients with Graves' disease clearly showed transient deterioration of the thyroid function during the treatment period, data from these two patients were separately investigated. Urinary levels of NAG in the remaining ten patients with Graves' disease before, 1, 3, 6 and 12 months after the treatment were 15.59 +/- 7.93 (SD), 8.96 +/- 6.82, 4.39 +/- 2.33, 3.46 +/- 2.24, and 3.63 +/- 2.38 U/g.creatinine (g.Cr.), respectively. Those obtained before, 1 and 3 months after the treatment were significantly higher than those of the controls (2.85 +/- 1.12 U/g.Cr.). Free thyroid hormone levels became normal or low 3 months after the treatment. The two Graves' patients mentioned above showed a transient increase in urinary NAG with concomitant changes in free thyroid hormone levels. Urinary NAG levels in the patients with subacute thyroiditis before, 2, 4, and 6 weeks after the treatment were 16.56 +/- 10.97, 6.76 +/- 2.79, 3.14 +/- 0.48 and 3.70 +/- 1.44 U/g.Cr., respectively. Those obtained before and 2 weeks after the treatment were significantly higher than those of the controls. Free thyroid hormones were normal 2 weeks after therapy. Urinary NAG in the patient with silent thyroiditis was 9.60 U/g.Cr. on the first visit and gradually decreased.(ABSTRACT TRUNCATED AT 250 WORDS)

Use of inactive beta-D-galactosidase for elimination of interference by anti-beta-D-galactosidase antibodies in immune complex transfer enzyme immunoassay for anti-thyroglobulin IgG in serum using beta-D-galactosidase from Escherichia coli as label.

A novel enzyme immunoassay (immune complex transfer enzyme immunoassay) for anti-thyroglobulin IgG using beta-D-galactosidase from Escherichia coli as label was reported previously. This immunoassay was highly sensitive in demonstrating anti-thyroglobulin IgG not only in all patients with Graves' disease and chronic thyroiditis but also in a large proportion of healthy subjects. However, the detection of anti-thyroglobulin IgG at low levels in some serum samples was difficult, probably due to the presence of anti-beta-D-galactosidase antibodies. In the present study, the use of inactive beta-D-galactosidase was tested for elimination of interference by anti-beta-D-galactosidase antibodies. Preincubation of serum samples with excess of inactive beta-D-galactosidase resulted in sufficiently low backgrounds to detect low levels of anti-thyroglobulin IgG with little effect on the dose-response of anti-thyroglobulin IgG. As a result, it was revealed that anti-thyroglobulin IgG was present in almost all healthy subjects as well as all patients with Graves' disease and chronic thyroiditis.

Serum angiotensin-converting enzyme. Alterations in hyperthyroidism, hypothyroidism, and subacute thyroiditis.

Angiotensin-converting enzyme (ACE) activity was measured in the serum samples of 247 patients with varying thyroid states. In 60 hyperthyroid patients, the mean (+/- SD) ACE level of 29.5 +/- 9.7 units/mL was higher than in all other groups. The serum ACE level was 17.0 +/- 5.1 units/mL in 129 euthyroid subjects and differed from the level of 13.9 +/- 5.1 units/mL observed in 34 hypothyroid patients. Twenty-four patients receiving exogenous thyroid hormone had elevated serum thyroxine values. Their mean serum triiodothyronine level was in the normal range, and the mean ACE level did not differ from the euthyroid mean. The mean serum ACE level fell from 30.8 to 17.4 units/mL in 35 hyperthyroid patients studied before and after therapy. In 12 hypothyroid subjects, the mean serum ACE level rose from 11.6 to 15.8 units/mL after thyroid hormone replacement. In eight of ten patients with transient hyperthyroidism (subacute thyroiditis or painless thyrotoxic thyroiditis), their highest ACE levels were observed in the hyperthyroid or transition phase and fell progressively with the lowest values being recorded during the hypothyroid or early recovery phases. Thus, ACE activity may respond to thyroid hormone, and interpretation of serum ACE levels may require knowledge of the patient's thyroid status. Serum ACE may be useful as a probe for exploring peripheral thyroid hormone action.

Histochemical and biochemical study on adenylate cyclase and 5'-nucleotidase activity in thyroid glands with normal and various thyroid diseases.

The adenylate cyclase and 5'-nucleotidase activity was measured biochemically in the thyroid glands from patients with various thyroid diseases in comparison with normal thyroid. The basal adenylate cyclase activity in normal thyroid was 159.3 p-moles cAMP/min./g tissue. The activity was elevated to 230% of basal with 20 mM NaF and 190% of basal with 100 mU/ml TSH. These values in chronic thyroiditis and Graves' disease were not significantly different from the values of normal thyroid. In adenomatous goiter, adenoma and carcinoma, the basal adenylate cyclase activity was significantly higher than that of normal thyroid. Parallel to the biochemical determination of both enzyme activities, the distribution of histochemically demonstrable adenylate cyclase and 5'-nucleotidase activity was described in the follicular cells with normal and various thyroid diseases. The reaction product of adenylate cyclase and 5'-nucleotidase activity was restricted to the plasma membrane of the follicular cells. However, the distribution and intensity of the adenylate cyclase reaction varied in each thyroid disease, except for the absence of reaction product in the basal plasma membrane. The lack of demonstrable adenylate cyclase activity in the basal plasma membrane suggests the possibility that the basal plasma membrane may not play an important role of TSH-reception.

Elevated serum angiotensin-converting enzyme in hyperthyroidism.

Serum angiotensin-converting enzyme was elevated in patients with hyperthyroidism (72 +/- 31 nmol/minute/ml, n = 12, p less than 0.001) but not in patients with hypothyroidism (38 +/- 3, n = 3) or thyroiditis (26, n = 1), and was positively correlated in 23 patients with serum thyroxine concentration (r = 0.60, p less than 0.01) and triiodothyronine resin uptake (r = 0.56, p less than 0.01). Triiodothyronine failed to enhance the synthesis of angiotensin-converting enzyme in rabbit alveolar macrophages or in human monocytes in culture, suggesting that the increased serum enzyme is a consequence of an effect other than increased angiotensin-converting enzyme synthesis. Hyperthyroidism should be considered in the evaluation of serum angiotensin-converting enzyme for the diagnosis and management of sarcoidosis.

Significance and function of multinucleate giant cells in de Quervain's thyroiditis.

Thyroid gland resections from 12 patients suffering from de Quervain's thyroiditis were investigated by light-, electron- and immunofluorescence-microscopy as well as enzymehistochemically . Several granulomas were serially sectioned. The giant cells of de Quervain's thyroiditis arise from the fusion of mononuclear precursors. They usually have ordered nuclei similar to the Langhans type. Histochemically they show the reactions of macrophages. Most enzyme activity is demonstrable in the 'working poles' of the giant cells abutting the residual thyroid colloid. We assume that as a result of polarization and orientation they are able to degrade the colloid in a direct manner. Sometimes they fuse to form syncytia which totally enclose the colloid.

Na-K-dependent ATPase in red cells and thyroid status.

The Relationship between ouabain-sensitive ATPase (Na-K ATPase) activity in erythrocytes and the thyroid status was studied in 36 patients with Graves' disease and 58 patients receiving L-thyroxine (T4) replacement therapy. Forty normal children served as control. Total ATPase activity in 4 untreated hypothyroid patients was significantly reduced (11.0 +/- 4.6 vs 17.3 +/- 4.1 micrograms-P/h/mg-protein, P less than 0.01), and Na-K ATPase was undetectable, both of which were normalized after 4 weeks of L-T4 therapy. Na-K ATPase in hyperthyroid patients was also decreased (0.9 +/- 0.8 vs. 4.0 +/- 2.7, P less than 0.01), but was gradually normalized after 3 months of euthyroid state. Clinically euthyroid children treated with L-T4 were divided into 2 groups with regard to Na-K ATPase activity, normal and low. Analysis of the possible factors producing this difference revealed that, in primary hypothyroidism, the factor appeared to be the endogenous T4 level, while in patients with dwarfism, the secretory capacity of TSH or TSH-releasing hormone (TRH) was contributory. Thus Na-K ATPase activity in red cells remains within the normal range after L-T4 replacement in the presence of a severe degree of primary hypothyroidism or in association with secondary or tertiary hypothyroidism. Other factors such as the L-T4 dose, duration of the therapy, serum T4 and T3 concentrations, were not significantly different in the two groups. These results indicate that (1) Na-K ATPase in red cells is decreased in hyper- or hypothyroid state, (2) restoration of normal activity requires 1-3 months of euthyroid period, and (3) it is a sensitive index of peripheral thyroid status over the preceding few months.

Ultrastructural findings in Hashimoto's thyroiditis and focal lymphocytic thyroiditis with reference to giant cell formation.

Ultrastructural findings in two cases of Hashimoto's disease and two cases of focal lymphocytic thyroiditis are reported. Stimulated thyrocytes, oncocytes and degenerating thyrocytes were observed in all cases. Multinucleated thyrocytes and epithelial pseudogiant cells were identified in Hashimoto's disease only. Infiltrating lymphocytes, plasma cells, monocytes and macrophages were present in all cases. The ultrastructure of germinal centres was similar to that seen in lymphatic organs. Giant cells of both intra- and extrafollicular localization were seen in Hashimoto's disease. Most of the giant cells were macrophage-derived. Two different ways of giant cell formation were identified: besides the familiar dissolution of plasma membranes of adjacent macrophages, another mechanism of fusion was observed. At sites of contact, peculiar membrane structures were developed and disintegration of plasma membranes occurred in parts adjacent to these structures. These are not identical to desmosomes and are different from Langerhans' granules. They probably represent special organelles for the initiation of cellular fusion.

Catecholamine-thyroid hormone interactions: II. Thyroid hormone and platelet MAO activity in patients with thyroid disorders.

Platelet monoamine oxidase (MAO) activity and serum thyroxine indices were determined in 62 children and adolescents currently undergoing medical treatment for various thyroid disorders. The platelet MAO activity of these patients was similar to that of control and contrast groups previously reported, and there were no differences when patients were grouped according to specific thyroid disorders. Estimated free thyroxine and total thyroxine levels were generally in the upper normal or slightly elevated range and were not significantly related to MAO activity.

[Diagnostic value of the determination of serum hexokinase activity in thyroid cancer and autoimmune thyroiditis]. / Diagnosticheskoe znachenie opredeleniia aktivnosti geksokinazy v s vorotke krovi u bol'nykh rakom shchitovidnoi zhelezy i autoimmunnym tireoiditom.

The blood serum hexokinase (HK) level was studied in 122 patients suffering from thyroid diseases. Relatively low level of HK activity (3.96 +/- 0.78 ME) was seen in the blood of patients with thyroid malignant tumor and its rise was observed in all the patients with non-tumor injuries of this organ: in nodular euthyroid goitre 1.38 +/- 0.25 ME; in diffuse-toxic goitre 0.83 +/- 0.67 ME. The blood serum HK activity (3.43 +/- 0.73 ME) of patients with autoimmune thyroiditis did not statistically differ from that of the patients suffering from thyroid cancer.

[The significance of acid phosphatase activity in the cytologic diagnosis of thyroid malignancy (author's transl)]. / Uber die Bedeutung der sauren Phosphatase für die Zytodiagnostik der Schilddrüsenmalignome.

The activity of acid phosphatase in thyroid cells obtained by aspiration biopsy with thin needle was studied. The results were compared with those, found by a biochemical method in the operation material of the same cases. It has been proved that there is a significant difference between malignant tumours with cytochemically mostly positive acid phosphatase reaction and colloid adenomas with mostly negative reaction. The results obtained by biochemical method showed significant quantitative differences between malignant and benign tumours. The pattern of acid phosphatase subunits (isoenzymes) in the sera of patients with thyroid malignancies were pathologic and similar to those found in mammary cancers. It can be supposed that the behavior of acid phosphatase in thyreocytes depends on same metabolic properties, perhaps on some process characteristic for malignancy, e.g. growth activity. Even this finding of cytochemically positive acid phosphatase reaction cannot be awaited to be quite specific, it has proved as a useful marker signalizing the possibility of malignancy.