Micromethod of human thyrocyte cultures for detection of thyroid-stimulating antibodies and thyrotrophin

1981 ◽  
Vol 97 (3) ◽  
pp. 369-375 ◽  
Author(s):  
G. Stöckle ◽  
R. Wahl ◽  
F.J. Seif
Keyword(s):  
Graves Disease ◽  
Nodular Goitre ◽  
Thyroid Stimulating Antibodies ◽  
Protein Binding Assay ◽  
Cell Dispersion ◽  
Competitive Protein Binding Assay ◽  
Competitive Protein Binding ◽  
Long Acting ◽  
Hyperthyroid Patients ◽  
Tissue Digestion

Abstract. The activation of adenylate cyclase of human thyrocytes in primary cell cultures and the release of cAMP into the medium are used to detect thyrotrophin (TSH) and thyroid-stimulating antibodies (TSAb) in sera of patients with Graves' disease. Tissue digestion and cell dispersion are performed using a neutral protease of Bacillus polymyxa (Dipase II), which harvests more vital thyrocytes than does trypsin. The cells show an enhanced response to stimulation. The efficeincy of the cell preparation and cultivation is increased by using microtest plates instead of culture flasks. By this minimization only 0.6–1.8 × 106 cells are needed to evalute a single sample. Cyclic AMP concentrations are measured directly in the supernatant culture medium by a competitive protein binding assay with charcoal separation. The minimal detectable dose of bTSH is about 10 μU/ml. With increasing doses of bTSH cAMP concentrations rise to a peak at about 50 to 100 mU/ml, beyond which there is a gradual decrease of cAMP indicating negative cooperativity in the activating mechanisms. The 'long-acting thyroid stimulator' effect of TSAb is reflected by a protracted increase of cAMP to its maximal value. All 41 sera of hyperthyroid patients with Graves' disease were TSAb-positive, whereas sera of patients with toxic nodular goitre and of euthyroid controls were TSAb-negative.

GRAVES' DIEASE AND HASHIMOTO'S THYROIDITIS IN MONOZYGOUS TWINS

1973 ◽  
Vol 72 (1) ◽  
pp. 18-24 ◽  
Author(s):  
Bruce S. Chertow ◽  
William J. Fidler ◽  
Bruce L. Fariss
Keyword(s):  
Hashimoto’S Thyroiditis ◽  
Monozygotic Twins ◽  
Hashimoto's Thyroiditis ◽  
Graves Disease ◽  
Thyroglobulin Antibodies ◽  
Protein Binding Assay ◽  
Competitive Protein Binding Assay ◽  
Competitive Protein Binding ◽  
High Concentrations ◽  
Monozygous Twins

ABSTRACT Monozygous twins and their family were studied. One twin had Graves' disease and the other had Hashimoto's thyroiditis. Their mother had Hashimoto's thyroiditis and their maternal grandmother and maternal great aunt had hyperthyroidism. Thyroid biopsies were obtained in each of the twins and showed changes of Graves' disease and Hashimoto's thyroiditis, respectively. High concentrations of anti-thyroid microsomal and anti-thyroglobulin antibodies were present by competitive protein binding assay, but LATS was absent in the twins and their mother. The occurrence of Graves' disease and Hashimoto's thyroiditis in monozygotic twins supports the concept of a common genetic aetiologic factor in the basic pathogenesis of these two diseases; however, the expression of this factor is variable, acquired, and, at least in part, genetically independent.

Adrenal function in wether lambs implanted with trenbolone acetate, ostradiol or trenbolone acetate plus oestradiol

1984 ◽  
Vol 1984 ◽  
pp. 149-149
Author(s):  
M. N. Sillence ◽  
R. G. Rodway ◽  
E. J. Redfern
Keyword(s):  
Analysis Of Variance ◽  
Time Correlation ◽  
Classical Analysis ◽  
Trenbolone Acetate ◽  
Protein Binding Assay ◽  
Treatment Models ◽  
Competitive Protein Binding Assay ◽  
Adrenal Response ◽  
Competitive Protein Binding ◽  
Long Acting

We have previously demonstrated that trenbolone acetate (TBA) treatment of female lambs and rats reduces both plasma glucocorticoid concentrations and the adrenal response to ACTH. It has been suggested that this reduction in the catabolic glucocorticoids may aid the anabolic effects of this agent.Four groups of 6 wether lambs were implanted with TBA (40 mg), TBA (35 mg) plus oestradiol (5 mg) or with a long-acting oestradiol implant (Compudose). Hourly blood samples were taken for 24 h periods just before and 10 days after treatment and Cortisol concentrations measured using a competitive protein binding assay.If the classical analysis of variance procedure is followed ignoring the time correlation within individual animals then the residual sum of squares will be inflated and significant effects can be missed. The correlation can be modelled by extending the analysis of variance to include a Fourier Analysis. For each Fourier cycle considerd the variation can be explained in terms of an overall model, separate treatment models and separate individual models.

RELIABILITY CRITERIA FOR SATURATION ANALYSIS OF STEROIDS BY COMPETITIVE PROTEIN BINDING

1970 ◽  
Vol 65 (1_Suppl) ◽  
pp. S61-S78 ◽  
Author(s):  
Billy D. Reeves ◽  
David W. Calhoun
Keyword(s):  
Protein Binding ◽  
Assay Method ◽  
Statistical Control ◽  
Protein Binding Assay ◽  
Competitive Protein Binding Assay ◽  
System 2 ◽  
Competitive Protein Binding ◽  
Single Method ◽  
Assay Design ◽  
Proper Perspective

ABSTRACT This communication is an attempt to delineate and define reliability criteria for saturation analysis of steroids by competitive protein binding assay. The discussion of these criteria evolved from three major considerations of assay method that help to place the ultimate criterion of accuracy in proper perspective. These major considerations are: 1) the measurement system, 2) the assay design and 3) the calculations and statistical control. Such an approach permits an evaluation, both relative and absolute, for a single method or for multiple methods.

TESTOSTERONE AND DIHYDROTESTOSTERONE CONCENTRATIONS IN THE FLUID MILIEU OF SPERMATOZOA IN THE REPRODUCTIVE TRACT OF THE BULL

1973 ◽  
Vol 74 (1) ◽  
pp. 186-200 ◽  
Author(s):  
Venkataseshu K. Ganjam ◽  
Rupert P. Amann
Keyword(s):  
Mass Spectrometry ◽  
Seminal Plasma ◽  
Reproductive Tract ◽  
Binding Assay ◽  
Rete Testis ◽  
Protein Binding Assay ◽  
Highly Sensitive ◽  
Competitive Protein Binding Assay ◽  
Competitive Protein Binding ◽  
Isolated Compound

ABSTRACT Total 17β-hydroxyandrogen concentrations were determined using a competitive protein binding assay, for bovine reproductive fluids. Rete testis fluid and cauda epididymal plasma were separated from the spermcontaining fluids obtained through cannulae from conscious bulls. Al-through the concentration of total 17β-hydroxyandrogens in rete testis fluid was similar (P > 0.05) to that in cauda epididymal plasma (25 and 19 ng/ml), both fluids contained higher (P < 0.01) androgen concentrations than seminal plasma, accessory sex gland fluid or serum from peripheral blood (3–5 ng/ml). However, since the amount of cauda epididymal plasma recovered was much less than for rete testis fluid (0.25 vs 35 ml/day), cauda epididymal plasma contained less than 1 % of the total 17β-hydroxyandrogens which entered the epididymis in rete testis fluid (5 vs 883 ng/day). Testosterone and/or dihydrotestosterone were isolated from the reproductive fluids by Sephadex LH-20 chromatography and quantified by a simple, specific and highly sensitive microassay. Dihydrotestosterone was found only in cauda epididymal plasma (14 ng/ml); identification of the isolated compound was confirmed by mass spectrometry. Dihydrotestosterone accounted for 52% of the 17β-hydroxyandrogens in cauda epididymal plasma while 23 % was testosterone. Testosterone represented 70 % of the 17β-hydroxyandrogens in rete testis fluid and 91 % of those in blood serum. Physiological implications of this shift in androgen balance are discussed.

A Simplified competitive protein-binding assay for 25-hydroxycalciferol

1976 ◽  
Vol 68 (2) ◽  
pp. 99-105 ◽  
Author(s):  
B. Garcia-Pascual ◽  
A. Peytremann ◽  
B. Courvoisier ◽  
D.E.M. Lawson
Keyword(s):  
Protein Binding ◽  
Binding Assay ◽  
Protein Binding Assay ◽  
Competitive Protein Binding Assay ◽  
Competitive Protein Binding

NSILA AND FOETAL GROWTH

1979 ◽  
Vol 90 (3) ◽  
pp. 534-543 ◽  
Author(s):  
U. E. Heinrich ◽  
D. S. Schalch ◽  
M. H. Jawadi ◽  
C. J. Johnson
Keyword(s):  
Gestational Age ◽  
Serum Growth Hormone ◽  
Premature Newborns ◽  
Foetal Growth ◽  
Protein Binding Assay ◽  
Competitive Protein Binding Assay ◽  
Term Newborns ◽  
Competitive Protein Binding ◽  
Cortisol Levels ◽  
Serum Gh

ABSTRACT Employing a sensitive competitive protein binding assay for NSILA (non-suppressible insulin-like activity), circulating levels of this somatomedin (SM) have been measured throughout pregnancy, at parturition, and in foetal and newborn sera. Acid-dissociable serum NSILA (mean ± sem) in 57 women was significantly higher during pregnancy (1106 ± 46 μU/ml), than in 11 adult non-pregnant control subjects (844 ± 22 μU/ml), but not correlated with week of gestation or with serum growth hormone (GH) or cortisol levels. At parturition, the NSILA concentration in 28 cord sera (598 ± 38 μU/ml) was significantly less than in the corresponding maternal sera (1039 ± 63 μU/ml). The NSILA levels in 23 premature newborns (370 ± 20 μU/ml) and 8 smallfor-gestational-age newborns (310 ± 46 μU/ml) were significantly less than in 33 term newborns (494 ± 18 μU/ml). Serum NSILA in 56 term and premature newborns exhibited a significant positive correlation both with gestational age and birth weight but not with serum GH or cortisol levels. These data suggest that the maternal-foetal growth-promoting system is a highly complex one in which NSILA levels both in maternal and foetal circulations appear to be under multifactorial control.

A Competitive Protein-Binding Assay for 25-Hydroxycholecalciferol

1975 ◽  
Vol 17 (2) ◽  
pp. 57-57
Author(s):  
Yoshiki Seino ◽  
Tsunesuke Shimotsuji ◽  
Shintaro Okada ◽  
Teisuke Hiejima ◽  
Chiiko Ikehara ◽  
...  
Keyword(s):  
Protein Binding ◽  
Binding Assay ◽  
Protein Binding Assay ◽  
Competitive Protein Binding Assay ◽  
Competitive Protein Binding ◽  
25 Hydroxycholecalciferol
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