Structural analysis of the thyrotropin receptor in four patients with congenital hypothyroidism due to thyroid hypoplasia.

Sporadic congenital hypothyroidism is most commonly caused by developmental abnormalities of the thyroid gland. More rarely, it is due to defects in gene products involved in the regulation of the hypothalamic-pituitary-thyroid axis or thyroid hormone synthesis. Loss of function mutations in the thyrotropin (TSH) receptor have been shown to result in resistance to biologically active TSH. In complete resistance to TSH, the thyroid gland is hypoplastic and unable to synthesize and secrete sufficient amounts of thyroid hormones. In partial resistance, referred to as euthyroid hyperthyrotropinemia, the size of the gland and the thyroid hormone levels are normal at the expense of an elevated TSH. Four patients with sporadic congenital hypothyroidism and properly located hypoplastic thyroid glands were included in this study. Serum TSH concentrations were 150 mU/L or higher, serum thyroglobulin levels were within normal limits (6.1 to 8.2 ng/mL; normal range: 2.1 to 32 ng/mL), and thyroid autoantibodies were absent. The coding region of the TSHbeta subunit gene, the TSH receptor gene, and exons 8 and 9 of Gsalpha were analyzed by direct sequencing and found to be normal in all patients. One patient was heterozygous for a G to A transition in the TSHbeta gene resulting in a substitution of alanine by threonine at position -7 of the signal peptide. This substitution was also found in her euthyroid father. In addition, Southern analysis of the TSH receptor gene excluded major structural alterations. These findings support previous reports that indicate that TSH resistance is genetically heterogeneous. In addition to mutations in the TSH receptor or the Gsalpha genes, other genetic defects can lead to an identical phenotype. These observations also suggest that TSH receptor mutations might be a relatively rare cause of congenital thyroid hypoplasia.

Autosomal recessive deficiency of combined pituitary hormones (except ACTH) in a consanguineous Brazilian kindred.

Familial hypopituitarism represents a clinically and genetically heterogeneous disorder. In a subset of these families, defects in Pit-I, a transcription factor essential for proper pituitary development have been identified as underlying molecular cause. These patients present extreme short stature, GH, PRL and TSH deficiency but intact ACTH, LH and FSH secretion. The pituitary is usually hypoplastic. In this report we describe a consanguineous family (the parents are first cousins) with thirteen siblings. Of the ten living siblings, four (two males and two females) have panhypopituitarism with severe growth failure. They had evidence of growth hormone, prolactin and gonadotropin deficiencies and developed central hypothyroidism late in life. ACTH secretion was normal. Bone age was retarded and dual-photon bone densitometry indicated severe osteoporosis. Combined provocative tests for pituitary hormones indicated blunted responses for GH, LH, FSH and a modest rise in serum PRL and TSH. A clonidine-test failed to induce pituitary GH response. A corticotropin-releasing factor (CRF) provocative test was conducted after 6 months without the use of prednisone with a normal ACTH response after CRF in the affected sibling. Plasma IGF-I and IGF-BP3 were below normal levels. Serum E2 (females) and serum testosterone (males) levels were very low. MRI evaluation of the pituitary indicated pituitary aplasia in all subjects. The phenotype described in this kindred is different from families reported with Pit-1 mutations. However, it resembles previously published kindreds with similar clinical and biochemical findings. The relative preservation of ACTH suggests a genetic defect early in pituitary gland development.

Autosomal recessive deficiency of combined pituitary hormones (except ACTH) in a consanguineous Brazilian kindred.

Familial hypopituitarism represents a clinically and genetically heterogeneous disorder. In a subset of these families, defects in Pit-I, a transcription factor essential for proper pituitary development have been identified as underlying molecular cause. These patients present extreme short stature, GH, PRL and TSH deficiency but intact ACTH, LH and FSH secretion. The pituitary is usually hypoplastic. In this report we describe a consanguineous family (the parents are first cousins) with thirteen siblings. Of the ten living siblings, four (two males and two females) have panhypopituitarism with severe growth failure. They had evidence of growth hormone, prolactin and gonadotropin deficiencies and developed central hypothyroidism late in life. ACTH secretion was normal. Bone age was retarded and dual-photon bone densitometry indicated severe osteoporosis. Combined provocative tests for pituitary hormones indicated blunted responses for GH, LH, FSH and a modest rise in serum PRL and TSH. A clonidine-test failed to induce pituitary GH response. A corticotropin-releasing factor (CRF) provocative test was conducted after 6 months without the use of prednisone with a normal ACTH response after CRF in the affected sibling. Plasma IGF-I and IGF-BP3 were below normal levels. Serum E2 (females) and serum testosterone (males) levels were very low. MRI evaluation of the pituitary indicated pituitary aplasia in all subjects. The phenotype described in this kindred is different from families reported with Pit-1 mutations. However, it resembles previously published kindreds with similar clinical and biochemical findings. The relative preservation of ACTH suggests a genetic defect early in pituitary gland development.

Osteopenia e tratamento com L-tiroxina / Osteopenia and treatment with L-thyroxine

Os hormônios tiróideos (T3 e T4) exercem nítido efeito no osso, como se pode comprovar pela perda de massa óssea que, usualmente, acompanha o hipertiroidismo näo tratado. A terapêutica convencional com L-tiroxina pode se acompanhar, em alguns pacientes, de hipertiroidismo tissular, o que, eventualmente, poderia condicionar maior risco para futura osteoporose. Recomendou-se, por tanto, que a terapêutica cim L-tiroxina fosse mais criteriosa, com reduçäo da dosagem diária. Mais recentemente, contudo, vários e importantes trabalhos científicos neste mesmo assunto indicaram que a terapêutica com L-tiroxina näo afeta substancialmente a massa óssea. A análise cuidadosa do que já foi publicado no assunto conclui que a possível açäo de perda de massa óssea por uso prolongado de L-tiroxina seria difícil de ser ignorada, mas que a magnitude deste efeito, possivelmente, é mínima e de conseqüências clínicas pouco significantes, sendo corrigíveis com maior ingestäo de cálcio e eventual terapêutica estrogênica na populaçäo feminina

[Osteopenia and treatment with L-thyroxine]. / Osteopenia e tratamento com L-tiroxina.

Thyroid hormones exert potent effects in bone as evidenced by bone loss which accompany untreated thyrotoxicosis. Conventional thyroxine replacement therapy may be associated with some degree of biochemical or tissue hyperthyroidism, increasing the risk for late onset osteoporosis. This has led to recommendations for reductions in L-T4 dosages. More recently, however, other reports have indicated that L-T4 therapy does not have adverse effects on bone mass. The overall analysis of both past and recent published reports has indicated that the evidence that conventional thyroxine therapy has adverse effects on bone mass is hard to ignore, although the magnitude of these effects is likely to be relatively small and closely related to estrogen deficiency.

Human thyroid tissue do not express thyroalbumin.

Thyroid tissue total RNAs from multinodular goiter (G2) and from hereditary goiter with defective Tg synthesis (JNA) were hybridized with a 5'albumin cDNA probe (F-47), a 3' albumin cDNA probe (B-44) and a thyroglobulin cDNA probe (phTgM3). JNA refers to tissue obtained from a patient with virtual absence of Tg in thyroid tissue and the presence of increased concentration of an albumin-like labeled protein in the thyroid. No hybridization signal was detected in both G2 and JNA with albumin probes at Northern Blot studies. Those results were confirmed by dot-blot analysis of total RNA where no hybridization signal was detected in G2 and JNA. To confirm that thyroid tissues do not express thyroalbumin total RNA from JNA and normal control thyroid tissue (C) were amplified by PCR using albumin and Tg primers. An expected fragment of 592 bp was observed in a human liver sample with the albumin primers. However JNA and C samples showed absence of an amplification product of the same size. We concluded that thyroid cells do not contain the albumin transcript. Albumin is probably taken up from circulation and iodinated by the thyroid follicular cell with subsequent release of iodoalbumin into the circulation.

Defective organification of iodide causing hereditary goitrous hypothyroidism.

We present a survey of the current state of knowledge about the prevalence of the syndrome involved in defective organification of iodide, and the mechanism of iodination and coupling catalyzed by the thyroid peroxidase (TPO) enzyme. A brief summary of the recent developments in molecular cloning of TPO and regulation of TPO gene expression is also included. Methods for purification of the enzyme and details about the assessment of TPO activity in tissue are briefly explained. The classification of defective organification of iodide is primarily based on the site of the biochemical defect, being quantitative (TPO absent) or qualitative (TPO structure, localization or apoenzyme are defectives). The presence of TPO inhibitors is also briefly described. The rare possibility of an absent source of peroxide (H2O2) causing defective iodide organification is discussed. Analysis of the 118 reported cases shows that the biochemical classification covers a spectrum of abnormalities and it is likely that further molecular biology studies will increase this heterogeneity as well as refining it. Genetic studies have suggested linkage between the TPO gene polymorphisms and the iodide organification defect and can be of importance for carrier detection and prenatal diagnosis. Neonatal screening for hypothyroidism is likely to expand the number of cases available for DNA analysis and possibly the molecular diagnosis. The importance of the mutations that would affect the histidine (His) residues in the translated protein was recently documented by the finding of a deletion removing part of exon 9 and thus also deleting a proximal His residue. The resulting TPO enzyme was inactive for iodide organification and coupling reaction. It is hoped that in time we will be able to expand our knowledge of the molecular diagnosis of the inborn errors of iodide organification.

Thyroid peroxidase in endemic goiter tissue.

Thyroid peroxidase (TPO) activity and TPO protein were analyzed in endemic goiter tissue under iodine deficiency and after iodine supplement. TPO was prepared from 9 endemic goiter tissues and 4 normal thyroid tissues by solubilizing enzyme with detergent. Four patients with endemic goiter received iodized oil injection 12 months before surgery. All patients had normal serum thyrotropin (TSH) and thyroid hormone levels before surgery. TPO activity was measured by iodinase assay and guaiacol assay. Endemic goiter TPO showed greater iodination activity than that of normal TPO (p less than 0.01). The guaiacol assay showed greater TPO activity in 6 of the 9 endemic goiter tissues than that of the normal tissue. Iodized oil treatment did not affect TPO activity or TPO proteins when compared with those in untreated endemic goiter tissues. TPO activity in endemic goiter tissue correlated with thyroid T4 5'-deiodinase activity and not with thyroid hormone content in thyroglobulin. Since thyroid T4 5'-deiodinase and TPO are under control of TSH, an increase in TPO activity in the presence of normal serum TSH may be explained by the increased sensitivity of endemic goiter tissue to TSH-one of the possible adaptation mechanisms of endemic goiter.

Dyshormonogenetic goiter: presence of an inhibitor of normal human thyroid peroxidase.

A dialyzable, thermostable inhibitor of normal thyroid peroxidase (TPO), with UV absorption maximum at 250-260 nm, was found in the digitonized, washed particulate fraction of two dyshormonogenetic goiters. No intrinsic TPO iodide-oxidation activity was detectable in either of these goiters, and their TPO iodination activity was below the method sensitivity threshold, even after dialysis. These findings could be explained by an absent or abnormal TPO associated with the synthesis of a TPO-inhibitor, or by the irreversible inhibition of a normal enzyme by the inhibitor.

Deficiency of superoxide dismutase in endemic goiter tissue.

Superoxide dismutase (SOD) activity and its concentration were measured in thyroid tissues obtained from patients with Graves' disease, Hashimoto's thyroiditis, differentiated thyroid cancer, and endemic goiter (before and after iodine supplementation) as well as in normal thyroid tissue (paranodular tissue) from patients with follicular adenomas. SOD activity was measured by pyrogallol assay in ethanol-chloroform extracts of the thyroid homogenates. The SOD concentration in the thyroid extract was measured as immunoreactive SOD by electroimmunoassay. Endemic goiter tissues (n = 10) contained significantly lower SOD activity [mean, 1.9 +/- 1.9 (+/- SD) vs. 7.5 +/- 3.9 ng purified SOD/micrograms DNA; P less than 0.02] and concentration (mean, 0.2 +/- 0.1 vs. 0.8 +/- 0.5 ng SOD/microgram DNA; P less than 0.01) compared with those of normal tissues. No other pathological thyroid tissues had such consistently low SOD levels. Lactate dehydrogenase activity, a marker of cytosolic enzyme, was not lower in endemic goiter tissues than in normal tissues, suggesting that both tissues possessed functioning cells capable of producing cytosolic enzyme. Thyroid tissue from endemic goiter patients previously treated with iodized oil injection also had low SOD activity and concentration. Western blot analysis indicated that SOD protein in the endemic goiter tissue did not differ from that in normal thyroid tissue. We conclude that there is deficiency of cytosolic SOD in endemic goiter tissue. Since the deficiency of cytosolic SOD causes more prolonged exposure to oxygen free radicals, the decrease in SOD might contribute to the degenerative changes frequently found in these tissues.

Towards the eradication of iodine-deficiency disorders in Brazil through a salt iodination programme.

Iodine-deficiency disorders have been a serious public health problem in Brazil because of the failure of a salt iodination programme established in 1953. The reasons for this failure were logistical, e.g., potassium iodide was not supplied to all salt-producers, iodination of salt was largely erratic, and part of the population at risk used only non-refined salt, which was not iodinated. In 1978 a task force was therefore formed to implement measures to eliminate iodine-deficiency disorders from the country. For this purpose, potassium iodate was distributed, free-of-charge, to all salt mills and an iodate dosing spray was supplied without cost to small salt producers. Also, regional laboratories for determining iodine in salt were set up, inspectors made regular visits to the salt mills, and samples of salt from commerce and from the producers were analysed. More than 90% of the samples contained 10-30 mg iodine per kg. In three typical areas of the country with endemic goitre the urinary excretion of iodine increased from an average of less than 40 mug iodine to 125 +/- 38 mug iodine per g creatinine. In conclusion, the salt iodination programme was a complete success and could serve as a model for other countries with a high prevalence of iodine-deficiency disorders.

Thyroid growth immunoglobulins in large multinodular endemic goiters: effect of iodized oil.

Iodized oil (IO) was administered to 10 goitrous patients recently emigrated to São Paulo (SP) from iodine deficiency areas and to 42 goitrous patients from 2 Brazilian chronic iodine deficiency regions, Loreto and Luziania (L). Thyroid growth-promoting immunoglobulin G (IgG) thyroid-stimulating antibody, serum thyroglobulin (Tg), TSH, and thyroid hormones were measured before and 1 yr after IO administration. In all patients there was a remarkable reduction of gland mass associated with a significant decrease (P less than 0.01) in both basal serum Tg and peak Tg levels after bovine TSH administration. The mean percent Tg increase after bovine TSH treatment was reduced to 82% above basal levels compared with 224% before IO. Mean serum TSH levels, elevated only in the L group [7.3 +/- 11 (+/- SD) microU/ml] decreased to the normal range after IO (2.5 +/- 2.1 microU/ml). Serum T3 and T4 concentrations did not change greatly. Tests for microsomal antibodies were negative before and after IO. IgG concentrates of serum obtained before and after IO were tested for their ability to stimulate incorporation of [3H]thymidine into DNA or to increase intracellular generation of cAMP in FRTL-5 cells. Thymidine incorporation activity was found in 8 of 10 patients from SP [316 +/- 37% (+/- SEM); range, 140-480%] and 25 of 42 patients in the L group (mean, 206 +/- 14; range, 120-500%) before IO. Stimulation of thymidine incorporation reflected true growth-promoting activity, as confirmed by experiments measuring cell number, was not accounted for by TSH in the preparation, and reflected IgG action because it was abolished by absorption with antihuman IgG. IgG from only 1 patient in group SP and 4 patients in group L stimulated intracellular production of cAMP in FRTL-5 cells. All patients except 1 in both groups had no IgG stimulation (less than 120%) of growth-promoting activity 1 yr after IO treatment. There was a significant positive correlation between thyroid growth-promoting activity and serum Tg concentrations (r = 0.58; P less than 0.001), but no significant correlation was found with other parameters (TSH, T4, and T3). We conclude that growth-promoting IgGs lacking ability to stimulate cAMP production may play a role in the large multinodular goiters due to chronic iodine deficiency.

Persistent pituitary resistance to thyroid hormone in congenital versus later-onset hypothyroidism.

The pituitary and peripheral responses to L-T4 and L-T3 therapy were studied in 12 patients with congenital goitrous hypothyroidism, in 10 patients with an ectopic thyroid and onset of hypothyroidism at 3-8 years of age, and in 6 patients with adult-onset hypothyroidism, after they had had their chronic thyroid hormone replacement therapy discontinued for 30 days. They were first treated with increasing L-T4 (0.1, 0.2 and 0.4 mg daily) followed by L-T3 (0.05 and 0.2 mg daily) after stopping thyroid medication for another month. Ten normal subjects were treated identically. In normal individuals the peak TSH, alpha, and TSH-beta response to TRH was significantly decreased with 0.1 mg L-T4 or 0.05 mg L-T3 daily and was suppressed with 0.2 and 0.4 mg L-T4 or 0.2 mg L-T3 daily; serum cholesterol and triglyceride decreased significantly with 0.2 or 0.4 mg L-T4 or 0.2 mg L-T3 daily; testosterone-estradiol binding globulin (TeBG) increased significantly at the same doses. In congenitally hypothyroid patients receiving 0.2 mg L-T4 daily, the mean peak TSH after TRH was 24 +/- 17 microU/ml, whereas in patients with an ectopic thyroid or adult-onset hypothyroidism the peak TSH was significantly less at 5.9 +/- 8.8 and 5.5 +/- 5.7 microU/ml, respectively. Only at the highest doses of L-T4 (0.4 mg/day) or L-T3 (0.2 mg/day) was the TSH response to TRH suppressed in the congenitally hypothyroid group. The alpha and TSH-beta subunit levels followed those of TSH.(ABSTRACT TRUNCATED AT 250 WORDS)

Serum thyroglobulin (Tg) stimulation with bovine TSH: a useful test for diagnosis of congenital goitrous hypothyroidism due to defective Tg synthesis.

Sixteen patients with congenital goitre were submitted to a bovine TSH stimulation test (bTSH), and serum thyroid hormones (T3, T4), TSH and thyroglobulin (Tg) were measured before and after bTSH injection. In 9 patients with an organification defect basal levels of Tg were normal (19.4 +/- 3.1 micrograms/l) rising after bTSH to a mean level +/- SEM of 37.3 +/- 4.1 micrograms/l. Six patients with defective Tg synthesis or release had a mean basal level of serum Tg of 8.7 +/- 1.9 microgram/l (mean +/- SEM) and failed to raise serum Tg concentrations after bTSH (mean +/- SEM: 10.4 +/- 2.1 micrograms/l). In both groups a modest although significant (P less than 0.05) change in serum thyroid hormones after bTSH was noted. We conclude that the bTSH test may be used to distinguish the group with defective Tg synthesis or release from other types of congenital goitre.

Congenital goitrous hypothyroidism: discordant systolic time intervals, pituitary and peripheral responses to high daily doses of T4 or T3 therapy.

Left ventricular performance was studied by a noninvasive technique through the measurement of the systolic time intervals (total eletromechanical systole, left ventricular ejection (LVET) time, preejection period (PEP) and PEP/LVET ratio (Systolic Quotient) in 8 young adults with congenital goitrous hypothyroidism. All subjects showed lengthening of PEP, shortening of LVET and an increased PEP/LVET ratio associated with low serum T3 and T4, an exaggerated TSH response to TRH, high levels of serum cholesterol, triglycerides and carotene. They were treated with increasing L-T4 at monthly intervals (100, 200 and 400 micrograms daily), followed by L-T3 (50 and 200 micrograms daily) after stopping medication for another month. Systolic time intervals and the systolic quotient promptly reversed to the normal range with physiologic L-T4 (100 micrograms) or L-T3 (50 micrograms) replacement, but the TSH peak response to TRH was still present and exaggerated. Further reductions of the systolic quotient occurred with 200 micrograms L-T4, but not with supraphysiological doses (400 micrograms L-T4 or 200 micrograms L-T3) of thyroid hormones. The highest dose of L-T3 (200 micrograms/day) induced a significantly lower mean systolic quotient than 400 micrograms L-T4 daily, while 5 patients still had a significant TSH response to TRH. This was interpreted as discordant pituitary and cardiac response to L-T3 and L-T4 therapy. Serum cholesterol and triglycerides were considered as very sensitive index of thyroid hormone peripheral action. These had a significant positive correlation with changes in the left ventricular performance. Serum carotene, although decreasing significantly with L-T4 or L-T3 treatment, had no significant correlation with the systolic quotient.(ABSTRACT TRUNCATED AT 250 WORDS)