1/8. Normal thyroglobulin and thyroperoxidase gene expression in thyroid congenital defective thyroglobulin synthesis.We have studied a member (JBM) of a family MO previously described, with congenital goiter, hypothyroidism, and presence of hyposialylated Tg in the follicular lumen. Other congenital goiters (MA and JNA) with virtual absence of Tg were studied similarly. The presence of apparently normal-sized Tg in JBM tissue was confirmed in the present study by radioimmunoassay, Sephacryl S300 column chromatography, immunoelectrophoresis, and SDS agarose gel electrophoresis. Dot blot hybridization analysis with Tg and TPO probes indicated that mRNA hybridization levels of JBM tissue were similar to control thyroid tissues. Congenital goiter tissues showed relatively lower TSH receptor mRNA content in comparison with normal thyroid tissues. dna was digested with five restriction endonucleases (Taq I, Eco Rv, Pvu II, Pst I, and Eco RI), and the results revealed polymorphisms previously described with the Tg gene. No significant differences in the TPO Pst I pattern were observed in comparison with control samples. We conclude that no major alterations of the Tg and TPO gene expression are detectable and that no significant deletions of these genes are present. The biochemical abnormality in the JBM Tg molecule may be a posttranslational error during the assembly of the protein.- - - - - - - - - - ranking = 1keywords = chromatography (Clic here for more details about this article) |
2/8. Differential levels of thyroid peroxidase and thyroglobulin messenger ribonucleic acids in congenital goiter with defective thyroglobulin synthesis.The biosynthesis of thyroid hormones requires iodide, thyroid peroxidase (TPO), thyroglobulin (Tg) and H2O2. We have studied two sisters with congenital large goiters and hypothyroidism. Perchlorate tests were negative. serum T3 and T4 were decreased, TSH was increased and Tg was within the lower limit of normal. Biochemical and molecular studies were performed on goiter samples obtained after surgery. Tg content in both tissues was negligible. paper chromatography of labeled iodocompounds showed a decrease in T4, and the presence of a pronase/pancreatin-resistant iodoprotein. TPO activity was normal in the tissues. Sephacryl S-300 gel filtration demonstrated labeled iodoalbumin-like protein and the absence of a Tg peak. Salting out studies of soluble protein fraction gave an abnormal pattern. Agarose gel electrophoresis showed the presence of an iodoalbumin-like protein and the absence of Tg in the tissues. This last finding was confirmed by immunoelectrophoresis. The Tg and TPO mRNAs levels were also analyzed. Dot-blot hybridization studies with pM5 (TPO cDNA) and phTgM2 (Tg cDNA) probes showed increased and decreased signals, respectively. The increase in TPO mRNA can be explained as a compensatory mechanism vis a vis an increase in serum TSH caused by decreased serum T3 and T4 due to the impairment in Tg mRNA. The Tg mRNA of both patients was further studied with four different probes covering 5' and 3' regions (phTgM1, phTgB1, phTgB2 and phTgB3). Hybridization was observed with all four probes, thus excluding a dramatic deletion defect. Northern transfer showed a clear signal of hybridization with the phTgB1 probe in the 8-9 Kb range. We may conclude that the biochemical and molecular abnormality of these patients is characterized by a decrease of Tg mRNA and of Tg translation.- - - - - - - - - - ranking = 1keywords = chromatography (Clic here for more details about this article) |
3/8. Lipid-rich cell thyroid adenoma: histopathology with comparative lipid analysis.A second case of the unique lipid-rich cell thyroid adenoma is described complemented by detailed lipid analysis. New observations were made. The cytoplasm of the tumour cells contained scattered, aggregated sudanophil crystals; under polarized light the frozen, unstained sections exhibited numerous birefringent lipid crystals; electron microscopy provided further evidence that the clear cell appearance was due to intracellular lipid droplets with scanty glycogen particles. Comparative lipid analysis by thin layer chromatography and high-pressure liquid chromatography (HPLC) revealed quantitative and qualitative differences in lipid composition of tumour cells when compared with goitre cells from normal thyroid gland and subcutane fat. Qualitative differences in triglyceride composition (by HPLC) between tumour cells and subcutaneous fat indicated that the fat accumulation in the follicle cells was not a result of simple storage, but an expression of altered intracellular lipid metabolism.- - - - - - - - - - ranking = 2keywords = chromatography (Clic here for more details about this article) |
4/8. Coexistence of autoantibody to human thyrotropin (TSH) and autoanti-idiotypic antibody to antihuman TSH antibody in a case with simple goiter.A 70-yr-old woman with simple goiter showed normal serum levels of T4, T3, free T4, TSH receptor antibody (TRAb) and increased TBG. Discrepancy in serum hTSH level was observed by different assay methods. Coexistence of both autoantibodies for hTSH and for anti-hTSH antibody were demonstrated by the reaction of the patient's antibody with both 125I-hTSH and 125I-anti-hTSH (monoclonal antibody; mAb). These two autoantibodies belong to the polyclonal immunoglobulin g (IgG). The autoantibody for hTSH recognized only beta-subunit of hTSH. Neither stimulating type of TRAb in Graves' disease nor blocking type of TRAb in primary hypothyroidism interfered with the binding of the patient's antibody to 125I-hTSH or 125I-anti-hTSH. Anti-idiotypic antibody (anti-ID antibody) for anti-hTSH antibody was purified by anti-hTSH antibody affinity chromatography. The binding reaction of 125I-anti-hTSH (mAb) with this anti-ID antibody could be inhibited by the unlabeled hTSH. This anti-ID antibody might represent the internal image of the nonbiological active site of TSH molecule, because of absence of thyroid stimulating activity. goiter in this patient may have occurred by the unbound TSH with IgG (free TSH) and the bound TSH with IgG, because TSH levels in both the whole serum and the IgG free serum (the unbound TSH with IgG) were decreased significantly by T4 treatment. Coexistence of these antibodies may participate in the autoimmune mechanism of an idiotype-anti-idiotype network.- - - - - - - - - - ranking = 1keywords = chromatography (Clic here for more details about this article) |
5/8. Normal level of thyroglobulin messenger ribonucleic acid in a human congenital goiter with thyroglobulin deficiency.Two siblings with congenital goiter were investigated from clinical, biochemical, and molecular biology standpoints. The association of clinical and biological hypothyroidism with undetectable levels of serum thyroglobulin (Tg) and the presence of iodohistidines in the urine suggested the diagnosis of defective Tg gene expression. This conclusion was confirmed by analysis of proteins present in goiter extracts. Only minute amounts of Tg-related material was detected by RIA (0.28 and 0.17 mg/g tissue compared to 80-100 mg/g in normal thyroid tissue), by sepharose 6B chromatography, and by sucrose density gradient centrifugation. Surprisingly, the goiters contained normal amounts of Tg mRNA. The size of the mRNA and the sequence organization of its first five exons also were normal. We conclude that no gross alteration of structure or transcription of the Tg gene was present in these patients. The results are compatible with a lesion affecting the mRNA sequence (point mutation, splicing error etc.), leading to defective translation or abnormal routing of the translation product through the membrane system of the cell. This latter hypothesis is supported by the extreme distension of the goiter endoplasmic reticulum found on electron microscopy.- - - - - - - - - - ranking = 1keywords = chromatography (Clic here for more details about this article) |
6/8. Hereditary congenital goitre with thyroglobulin deficiency causing hypothyroidism.The thyroid glands of two hypothyroid goitrous siblings aged 13 and 14 and of a 21-year-old hypothyroid goitrous female were examined. In all three patients a very high thyroid uptake of iodide was observed in the presence of a negative perchlorate discharge test. An abnormally high serum protein bound iodine (12.9-20.0 micrograms/dl) and low serum T4 concentration suggested the presence of increased serum levels of iodoalbumin. Surprisingly, serum T3 levels were normal or low normal (80-220 ng/dl) in several determinations. Basal serum TSH was elevated and an exaggerated TSH response was observed after TRH. serum thyroglobulin was undetectable in one patient, low normal in another and in the normal range for the third one. Except for the patient with undetectable Tg the two other subjects slightly increased the serum Tg levels after a bovine TSH injection. plasma chromatography after a tracer dose of 125I disclosed only minute amounts of T3 T4 and MIT DIT. Studies performed in the homogenized thyroid tissues indicated that these goitrous glands had pronounced decrease of immunoreactive thyroglobulin. The total amount of Tg-like proteins (RIA) in the thyroid soluble protein extract was only 16-122 micrograms/g (normal: 50-70 mg/g of tissue). Ultracentrifugal studies were unable to demonstrate the presence of mature (18-20S) thyroglobulin. Only one peak (3.6-4.1S) was obtained in the pooled soluble proteins supernatants. hydrolysis of the homogenates indicated, by subsequent column chromatography, very low relative concentrations of iodotyrosines and iodothyronines and that a relatively large amount of iodide remained associated with subcellular proteins and undigested. The predominant histological pattern was of the intermediary differentiated adenoma type, microfollicular or fetal, with several atypical features and capsular invasion which may suggest malignant change. We conclude that a defective Tg export from the cell to the lumen or an anomaly in the structural gene leading to inadequate translation of Tg mRNA finally results in deficient storage of normal, mature Tg in the colloid with subsequent goitrous hypothyroidism.- - - - - - - - - - ranking = 2keywords = chromatography (Clic here for more details about this article) |
7/8. Familial and sporadic thyroglobulin deficiency with goitre and hypothyroidism.The thyroid proteins and iodocompounds were analysed in the thyroid tissue of 4 patients with nontoxic goitre. Subtotal thyroidectomy was performed for tracheal compression. The thyroid components were labelled with a trace amount of 125I before operation. One patient had congenital goitre and hypothyroidism with cretinoid features. Three other patients belonged to the same family. Two had congenital goitre, one of them with subclinical and biochemical hypothyroidism. There was a range of thyroglobulin (TG) deficiency ranging from virtual absence of TG in the most affected patient to 17% of normal in the least affected one. There seemed to be an inverse relationship between TG content and clinical signs. Also with decreasing TG more iodocompounds were found in the 3-8 S region on gradient centrifugation. In the most affected patient all the radioactivity was in the 3-8 S region, in the least affected one it was all found in the 19 S and 27 S regions. The other patients had an intermediate pattern. The 3-8 S fraction contained albumin, IgG and some material which reacted like TG on immunoelectrophoresis except for the least affected patient. The iodine content was normal whereas the iodination of TG was low-normal or low. Iodotyrosines and iodothyronines were found in all glands analysed on column chromatography but only in the most heavily affected patient did the ratio iodotyrosines/iodothyronines seem to be elevated. Evidently TG was not necessary for hormone formation in this gland but the efficiency of the matrix seemed not to suffice for normal hormone production.- - - - - - - - - - ranking = 1keywords = chromatography (Clic here for more details about this article) |
8/8. Increased peroxidase activity in Pendred's syndrome with hypothyroidism.An 8-year-old boy with goiter and bilateral nerve deafness had a 46% discharge of radioiodine after thiocyanate administration. He was clinically euthyroid. Although the serum total T4 was low (2.4 mug/100 ml) and TSH was significantly high (181 muU/ml), the serum total T3 was normal (152 ng/100ml). It was considered that the increased release of TSH by the feedback mechanism in response to the low T4 resulted in a quite normal level of serum T3. The thyroid gland demonstrated a low stable iodine content, an increase in MIT/DIT ration and a decrease in iodothyronine. The thyroglobulin behaved normally in Sephadex G-200 chromatography and immunoreaction. Thyroid tissue exhibited increased peroxidase activity as measured by I3 formation. Increased peroxidase activity may be related to the observed increase in serum level of TSH.- - - - - - - - - - ranking = 1keywords = chromatography (Clic here for more details about this article) |