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1

Astereological analysis of FRTL-5 cells as an effect of thyrotropin and irradiation

100%
Stiblar-Martincic D., Cor A., Pajer Z.
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issue 3-4
173-177
EN
FRTL-5 cell line is a cloned epithelial follicular cell line from Fischer rat thyroids. This cell line expresses many of the thyroid differentiated markers in vitro. Their growth and function depend on thyrotropin (TSH) as the main regulatory hormone. In this astereological analysis, the authors concentrate on FRTL-5 nuclei as the most vulnerable part of the cell. Using morphometrical variables, they wished to discover the morphologically identifiable sign of transformation of FRTL-5 cells after irradiation and to study the effect of different TSH concentrations. FRTL-5 cells were grown in a medium of 4 different concentrations of TSH (0, 0.1, 1, 10 mU/ml) and irradiated with 0 Gy, 2 Gy, and 4 Gy. The results showed that the nuclear-cytoplasmic ratio decreases after irradiation with doses of 4 Gy or if TSH was included in the medium. The nuclear maximum diameter of FRTL-5 cells increased with higher concentrations of TSH more obviously after irradiation with 4 Gy than with 2 Gy. On the basis of astereological analysis, it was concluded that different concentrations of TSH and irradiation exert an effect especially upon FRTL-5 cell nuclei. The possible transformation of FRTL-5 cells after culturing in TSH medium and after irradiation could be confirmed by injection into an animal of the Fischer strain.
2

A mouse monoclonal antibody to the TSHR

100%
Stankowiak-Kulpa H., Sanders J., Depraetere H., Jeffreys J., Evans M., Richards T., Furmaniak J., Smith B. R.
Archivum Immunologiae et Therapiae Experimentalis
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2005
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vol. 53
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issue 4
345-351
EN
Mouse monoclonal antibodies (mAbs) with the ability to inhibit thyrotropin (TSH) binding to the TSH receptor (TSHR) are useful tools to study TSH-TSHR interaction. The 3C3 mAb we produced was found to inhibit binding of TSH to human (h)TSHR but not to porcine (p)TSHR. Purified 3C3 immunoglobulin G (IgG) and its antibody-binding framgnet were prepared using standard methods and their ability to inhibit TSH binding to hTSHR or pTSHR was analyzed using a coated tube assay. The TSHR epitope reactive with 3C3 IgG was determined using Western blotting, ELISA based on peptides corresponding to the TSHR sequence, and the SPOT synthesis technique. RNA was isolated from 3C3 hybridoma cells and the mAb variable (V) region genes were sequenced and analyzed. 3C3 mAb had a 1x108 l/mol binding affinity to the hTSHR as assessed by Scatchard analysis. 3C3 reacted with the hTSHR region between amino acids (aa) 212-230, and two aa differences were found between the corresponding regions in the hTSHR and pTSHR. The light chain (LC) genes of 3C3 were derived from the Vk21 germ line (97.6% homology) and Jk2 genes. The heavy chain (HC) genes were from the V130 germ line (94.6% homology) combined with a D gene (not identified) and JH3 gene. The replacement/silent mutation ratios of 6.0 and 6.5 for the LC and the HC V regions, respectively, indicated that 3C3 underwent antigen-driven maturation. Mouse mAbs of this type should be useful in studying the interactions between the TSHR, TSH, and mAbs in more detail.
3

Adrenal steroids act as inhibitory modulators of thyrofollicular cell morphology and proliferation in neonatal chicks (Gallus domesticus)

88%
Ganguli S., Bandyopadhyay R., Chakraborty S.
Folia Biologica
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2001
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vol. 49
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issue 3-4
259-263
EN
In the present study the effects of adrenal corticoids, both natural and synthetic, namely cortisol and dexamethasone respectively, was observed on the thyroid gland cell morphology and proliferation in neonatal male chicks (Gallus domesticus). Cortisol was injected at a dose of 4 mg/100 g body weight and dexamethasone at a dose of 1 mg/100 g b.w. subcutaneously daily for fifteen consecutive days. The control birds were similarly injected with normal saline at a daily dose of 0.2 ml per bird for the same time period. The results indicated that both cortisol and dexamethasone caused a significant decrease in thyrofollicular cell height. On the contrary, a significant increase in the ratio of the follicular diameter to the number of nuclei per follicle i.e. D/N value was observed in both cortisol and dexamethasone treated chicks. It was also observed that both cortisol and dexamethasone induced suppression of mitotic activity, as evidenced from a significant decrease in mitotic percentage compared with the control chicks. The present authors? studies thus indicate that adrenal corticoids act as inhibitory modulators of thyroid follicular activity as regards karyomorphology and cell proliferation.
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