scholarly journals Improved assay method for activity of thyroid peroxidase-catalysed coupling of iodotyrosine residues of thyroglobulin utilizing h.p.l.c. for analysis of iodothyronines

1989 ◽  
Vol 262 (1) ◽  
pp. 209-214 ◽  
Author(s):  
T Ohmori ◽  
O Tarutani ◽  
T Hosoya
Keyword(s):  
Ethyl Acetate ◽  
Peroxidase Activity ◽  
Sodium Cholate ◽  
Human Thyroid ◽  
Assay Method ◽  
Thyroid Peroxidase ◽  
Graves Disease ◽  
Thyroid Glands ◽  
Hydrolysis Of

The coupling of iodotyrosine residues of thyroglobulin (Tg) catalysed by thyroid peroxidase (TPO) has scarcely been studied with respect to the TPO of abnormal human thyroid glands. The present paper proposes a rapid and convenient assay method applicable for determining the coupling activity of a sample of less than 500 mg from each patient's thyroid. The main characteristics of the method are as follows: (i) mitochondrial/microsomal fractions of thyroid glands were treated with sodium cholate plus trypsin, and the supernatants obtained by ultracentrifugation were directly used for the assay of coupling and peroxidase activity of TPO; (ii) the formation of iodotyrosine residues catalysed by TPO was performed by using chemically iodinated Graves'-disease Tg containing 41 iodine atoms per molecule and with a high iodotyrosine and a low iodothyronine content; (iii) newly synthesized iodothyronine residues (thyroxine, 3,5,3′-tri-iodothyronine, and 3,3′,5′-tri-iodothyronine) were analysed by h.p.l.c. after hydrolysis of Tg with proteinases and extraction of iodothyronines with ethyl acetate.

scholarly journals A rapid assay of human thyroid peroxidase activity

2020 ◽  
Vol 62 ◽  
pp. 104662 ◽  
Author(s):  
Hongyan Dong ◽  
Marlena Godlewska ◽  
Michael G. Wade
Keyword(s):  
Peroxidase Activity ◽  
Human Thyroid ◽  
Thyroid Peroxidase ◽  
Rapid Assay ◽  
Human Thyroid Peroxidase

Thyroid peroxidase and thyroglobulin expression in normal human thyroid glands

1998 ◽  
Vol 9 (4) ◽  
pp. 333-338 ◽  
Author(s):  
Marcus A. Lima ◽  
Valéria A. Gontijo ◽  
Fernando C. L. Schmitt
Keyword(s):  
Human Thyroid ◽  
Thyroid Peroxidase ◽  
Thyroid Glands ◽  
Normal Human

Intrathyroidal cytokine gene expression profiles in autoimmune thyroiditis

1994 ◽  
Vol 141 (2) ◽  
pp. 309-315 ◽  
Author(s):  
R Paschke ◽  
F Schuppert ◽  
M Taton ◽  
T Velu
Keyword(s):  
Autoimmune Thyroiditis ◽  
Hashimoto’S Thyroiditis ◽  
Thyroid Tissue ◽  
Human Thyroid ◽  
Hashimoto's Thyroiditis ◽  
Graves Disease ◽  
Thyroid Glands ◽  
Normal Human ◽  
Ifn Γ ◽  
Cytokine Mrnas

Abstract Cytokines are thought to mediate the initiation and perpetuation of autoimmune thyroiditis. However, this concept is mainly based on in vitro findings and to date only interleukin (IL)-6 and interferon-γ (IFN-γ) have been detected in Graves' disease in vivo. The cytokine pattern produced by T-helper (Th) cells has important regulatory effects on the nature of the immune response. We therefore determined these cytokine mRNAs in Graves' disease and Hashimoto's thyroiditis. RNA was extracted by cesium chloride gradient centrifugation from the thyroid tissue of 12 patients undergoing thyroid resection for Graves' disease and from two patients being treated for Hashimoto's thyroiditis. Two patients with parathyroid adenomas and one patient with a goiter were used as controls. RNA was also extracted from normal human thyroid epithelial cells in primary culture. The cDNAs were prepared by reverse transcription and amplified for IL-2, -4, -5, -6 and -10 and IFN-γ by polymerase chain reaction. All the cytokine mRNAs were detected in the Hashimoto's thyroid glands in large quantities. Six of the 12 Graves' disease thyroid glands showed, when compared with controls, an increased accumulation of transcripts for: IFN-γ, IL-2, -4 and -10 or IL-2, -4 and IFN-γ or IL-2 and IFN-γ or IFN-γ alone, each in one case or IL-2 alone in two cases. These cytokine profiles were not representative of a Th1 or Th2 phenotype. Increased amounts of cytokine mRNA in thyroid glands from Graves' disease patients were mostly associated with high microsomal antibody titres and/or prominent intrathyroidal lymphocytic infiltration. IL-6 and/or IL-10 mRNAs were detectable in all Graves' disease thyroid glands and in control thyroid tissue. IL-10 mRNA was not detectable in normal human thyroid epithelial cells in primary culture. Graves' disease and Hashimoto's thyroiditis clearly differ with respect to the number of positive intrathyroidal cytokine mRNAs and their levels. The different cytokine patterns in Graves' disease and in Hashimoto's thyroiditis could reflect the clinical spectrum of autoimmune thyroiditis which is characterized by thyroid tissue destruction and/or thyroid autoantibody production. These data suggest that the course of autoimmune thyroiditis is regulated by the interplay of several cytokines. Journal of Endocrinology (1994) 141, 309–315

Thyroid peroxidase expression in diseased human thyroid glands

1999 ◽  
Vol 10 (3) ◽  
pp. 223-228 ◽  
Author(s):  
Marcus A. Lima ◽  
Valéria A. Gontijo ◽  
Marlene C. Santos ◽  
Fernando C. L. Schmitt
Keyword(s):  
Human Thyroid ◽  
Thyroid Peroxidase ◽  
Thyroid Glands

scholarly journals Increased Expression of the Na+/I− Symporter in Cultured Human Thyroid Cells Exposed to Thyrotropin and in Graves’ Thyroid Tissue*

1997 ◽  
Vol 82 (10) ◽  
pp. 3331-3336 ◽  
Author(s):  
Tsukasa Saito ◽  
Toyoshi Endo ◽  
Akio Kawaguchi ◽  
Masato Ikeda ◽  
Minoru Nakazato ◽  
...  
Keyword(s):  
Thyroid Tissue ◽  
Human Thyroid ◽  
Thyroid Peroxidase ◽  
Graves Disease ◽  
Intracellular Camp ◽  
Normal Thyroid Tissue ◽  
Thyroid Cells ◽  
Normal Thyroid ◽  
Hormone Synthesis ◽  
Messenger Ribonucleic Acid

Abstract The Na+/I− symporter (NIS) is important in hormone synthesis in the thyroid gland. NIS activity, as reflected by I− uptake, was increased by TSH (1 mU/mL) or forskolin (10μ mol/L) in primary cultured human thyroid cells. Northern blot analysis revealed that incubation of these cells with TSH or forskolin for 24 h increased the abundance of NIS messenger ribonucleic acid (mRNA) 2.3- and 2.5-fold, respectively. Immunoblot analysis revealed 2.7- and 2.4-fold increases, respectively, in the amount of NIS protein after 48 h, suggesting that elevated levels of intracellular cAMP induced the expression of NIS in human thyrocytes. We then studied the levels of NIS mRNA and protein in Graves’ thyroid tissue and found that the amount of NIS mRNA in thyroid tissue from individuals with Graves’ disease (n = 5) was 3.8 times that in normal thyroid tissue (n = 5). The abundance of NIS mRNA was significantly correlated with that of thyroid peroxidase or thyroglobulin mRNAs, but not with that of TSH receptor mRNA, in the Graves’ and normal thyroid tissue specimens. The amount of NIS protein was also increased 3.1-fold in Graves’ thyroid tissue compared with that in normal thyroid tissue. The increased expression of NIS may thus contribute to the development of Graves’ disease.

An Improved Assay Method for Thyroid Peroxidase Applicable for a Few Milligrams of Abnormal Human Thyroid Tissues1

1985 ◽  
Vol 98 (3) ◽  
pp. 637-647 ◽  
Author(s):  
Toichiro HOSOYA ◽  
Ikuro SATO ◽  
Yoshio HIYAMA ◽  
Hirokatsu YOSHIMURA ◽  
Hiroo NIIMI ◽  
...  
Keyword(s):  
Human Thyroid ◽  
Assay Method ◽  
Thyroid Peroxidase

Microparticle-enhanced nephelometric immunoassay of anti-thyroid peroxidase autoantibodies in thyroid disorders

1994 ◽  
Vol 40 (3) ◽  
pp. 442-447 ◽  
Author(s):  
A A Harchali ◽  
P Montagne ◽  
J Ruf ◽  
M L Cuillière ◽  
M C Bene ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Hashimoto Thyroiditis ◽  
Human Thyroid ◽  
Thyroid Peroxidase ◽  
Graves Disease ◽  
Thyroid Disorders ◽  
Autoimmune Thyroid Diseases ◽  
Autoimmune Thyroid ◽  
Human Thyroid Peroxidase ◽  
Antigenic Domains

Abstract Crude thyroid peroxidase extracted from human thyroid microsomes was covalently bound onto polyacrylic and polyfunctional copolymerized microparticles. We observed agglutination of the thyroid peroxidase-microparticle conjugate with 13 monoclonal antibodies (mAbs) specific for epitopes on four different antigenic domains of human thyroid peroxidase (TPO; EC 1.11.1.7), after addition of anti-mouse immunoglobulins. We quantified agglutination by measuring with a specially designed nephelometer the light scattered by the conjugates. This allowed us to develop a microparticle-enhanced nephelometric immunoassay for human anti-TPO autoantibodies (aAbs) with defined epitopic specificity, based on the ability of aAbs to inhibit mAb-induced agglutination. Applied to patients with autoimmune thyroid diseases, this assay confirmed the polyclonality of anti-TPO aAbs and their preferential reactivity toward epitopes located on the A and B antigenic domains of the TPO molecule. The same specificities seem to be present in patients with Hashimoto thyroiditis or Graves disease.

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