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PubMed1975

Bovine serum thyroxine, prolactin, growth hormone, glucocorticoid, and thyroxine binding globulin response to thyroprotein.

Twenty-one lactating Holstein cows were assigned randomly to groups of seven and received: (1) no treatment; (2) 15 g thyroprotein daily for 5 wk; or (3) 15 g thyroprotein daily for 13 wk. Combined averages for serum thyroxine of cows in thyroprotein groups increased linearly from a baseline of 54 ng/ml to a peak of 135 ng/ml at 6 days after thyroprotein feeding was begun. Serum thyroxine then decreased to approximately 80 ng/ml at 23 days of thyroprotein feeding which was maintained during the remainder of the feeding period. Following thyroprotein withdrawal from the diet serum thyroxine concentration decreased to 24 ng/ml at 6 days. Neither serum prolactin, growth hormone, nor total glucocorticoids were affected by thyroprotein feeding or withdrawal. Average milk production of cows in the control group decreased linearly with time. Milk production of cows fed thyroprotein for 5 wk averaged 2.2 to 3.3 kg/day more than cows in the control group and that of cows fed thyroprotein for 13 wk averaged .95 to 2.5 kg/day more than controls for 60 days, but between 60 and 91 days it was .9 kg/day less than controls.

Animals↗
PubMed1990

Differentiating various abnormalities of thyroxin binding to serum proteins by radioelectrophoresis of thyroxin and immunoassay of binding proteins.

Using the simple method of protein analysis described here, we could identify thyroxin (T4)-binding-protein abnormalities in euthyroid patients with hyperthyroxinemia or hypothyroxinemia. Serum incubated with [125I]thyroxin was analyzed by agarose gel electrophoresis, with bromphenol blue staining of protein. The relative distribution of radioactive T4 was determined for each binding protein--thyroxin-binding globulin (TBG), transthyretin, albumin, and T4-binding immunoglobulin (when present)--and the mass of T4 bound to each was determined. We also used sensitive immunoassays to quantify TBG, transthyretin, and albumin concentrations, then calculated the mass of T4 (as determined by electrophoresis) bound per unit mass of the respective binding protein. When the concentration of binding proteins was altered (e.g., TBG excess or TBG deficiency), the T4 binding/mass ratio for each protein remained within the expected range; but when the functional affinity of a binding protein was altered--as in dysalbuminemic hyperthyroxinemia and in low-T4 nonthyroidal illness--this ratio was abnormal. This procedure can be used to help identify TBG excess, TBG deficiency, dysalbuminemic hyperthyroxinemia, prealbumin-associated hyperthyroxinemia, variant TBG with reduced affinity for T4, euthyroid sickness, and T4-binding autoantibodies.

Autoantibodies↗
PubMed1987

Clinical evaluation of a thyroxine binding globulin assay in calculating a free thyroxine index in normal, thyroid disease, and sick euthyroid patients.

As assay for thyroxine binding globulin (TBG) that measures the thyroxine binding capacity of TBG and other proteins was used to calculate a free thyroxine index (FTI) in 108 consecutive thyroid patients, 2,559 normal patients, and 152 sick euthyroid patients. The TBG assay compared favorably with the triiodothyronine (T3) uptake test in producing a FTI for the clinical evaluation of thyroid patients. In addition, it did not suffer the disadvantages inherent in assays specific for only TBG. In the sick euthyroid population, the TBG assay produced an FTI that was more consistent with the clinical evaluation than did the T3 uptake test.

Humans↗
PubMed1988

Thyroxin binding by human serum albumin after denaturation of the thyroxin-binding globulin in familial dysalbuminemic hyperthyroxinemia.

Abnormal binding of thyroxin (T4) to serum albumin of subjects with familial dysalbuminemic hyperthyroxinemia (FDH) is generally demonstrated by the T4-loaded charcoal uptake test, with T4 added in excess (0.1 mmol/L) to accentuate T4 binding to albumin in FDH. I describe a binding study involving T4 tracer in which thyroxin-binding globulin is denatured in samples by treatment with mild acid at pH less than 3.0. The tracer is bound to the serum albumin and, to a greater extent, to the FDH albumin, because the binding by thyroxin-binding prealbumin is blocked by barbital buffer. The result of the [125I]T4 binding to the albumin is expressed as a T4 binding index, based on results for pooled sera from patients with normal thyroid function as a reference. The mean index in FDH was 4.08 (SD 0.92, n = 5); in hypoalbuminemia, 0.66 (SD 0.18, n = 8); in normal subjects, 1.00 (SD 0.11, n = 20). This albumin-binding index enables the rapid and unequivocal diagnosis of subjects with FDH, without the addition of unlabeled T4.

Charcoal↗
PubMed1988

One-step chemiluminescent immunoassay of free thyroxin with acridinium-ester-labeled thyroxin evaluated and compared with a two-step radioimmunoassay.

This new one-step chemiluminescent immunoassay of free thyroxin (FT4) involving a thyroxin-immunoglobulin conjugate labeled with acridinium ester (Magic Lite System; Ciba Corning Diagnostics Corp., Medfield, MA) is rapid (one 1-h incubation), requires two calibrators per run, and takes 10 s per sample for the quantification step. Analytical performances were excellent: within- and between-run CVs of less than 10% in the working range, no significant effect of hemolysis, bilirubin, or lipemia, and no significant interaction between the conjugate and the thyroxin-binding proteins. Magic Lite results (y) correlated well with those obtained by the Sclavo (x) two-step radioimmunoassay (Sclavo, Siena, Italy): y = 1.35x + 1.32 (r = 0.94, n = 267, P less than 0.001, Sxy = 6.29). Clinical sensitivities (diagnostic efficiencies) for hypothyroidism and hyperthyroidism were 0.91 and 0.98 for normal interval limits of 12 and 21.5 pmol/L (95% confidence interval). Magic Lite results in situations where patient therapy, treatment, or unusual conditions can result in a lack of correlation between the clinical status and the FT4 values were qualitatively the same as those obtained by the Sclavo assay.

Adult↗
PubMed1983

Improved dialysis method for free thyroxin in serum compared with five commercial radioimmunoassays in nonthyroidal illness and subjects with abnormal concentrations of thyroxin-binding globulin.

We describe a method for free thyroxin (FT4) in serum, based on radioimmunoassay of T4 in serum dialysate, FT4(D). The method is analytically accurate, sensitive, reproducible, and suitable for routine use in the clinical laboratory. We compared results by this method with those obtained with five commercial FT4 assays (Amerlex, GammaCoat, ImmoPhase, LiquiSol, Spiria) and the free T4 index (FT4I). In several of the patients with nonthyroidal illness FT4(D) was above normal and FT4 as measured with the commercial assays was subnormal. In the third trimester of pregnancy all the FT4 methods gave decreased mean values, though the decreases were of various magnitudes: FT4(D) 32%, Amerlex 44%, GammaCoat 40%, Spiria 19%, LiquiSol 16%, and ImmoPhase 13%, respectively. FT4I was significantly higher than normal during late pregnancy. In women taking oral contraceptives the mean FT4I was increased, but there was no significant effect on the results obtained with the various FT4 methods. All subjects with hereditarily high or low serum thyroxin-binding globulin had normal FT4(D) and abnormal FT4I. Amerlex, ImmoPhase, and LiquiSol misclassified some of the subjects with hereditarily abnormal thyroxin-binding globulin.

Adult↗
PubMed1981

Simultaneous determination of free thyroxine and capacity of thyroxine-binding globulin.

A simple method is described for the simultaneous determination of capacity thyroxine-binding of globulin (TBG) and free thyroxine concentration (FT4). The ratio of bound to free T4 (B/F) is first determined for two total-T4 concentrations using a Sephadex G-25 competitive-binding technique. TBG capacity and FT4 can both the calculated assuming a known value of affinity constant of TBG. The method is linear over a twenty-fold serum dilution. FT4 calculated is identical to that calculated using the method of Irvine. TBG capacity is shown to be linearly correlated to TBG concentration as determined by radioimmunoassay and is consistent with a molecular weight of 69,000 Daltons and one T4 binding site per molecule. FT4 is found to correlate with the free thyroxine index in a complicated way, depending on the degree of TBG saturation.

Binding, Competitive↗
PubMed1995

Indirect estimation of thyroid hormone-binding proteins to calculate free thyroxine index: comparison of nonisotopic methods that use labeled thyroxine ("T-uptake")

There are many alternative ways of estimating free thyroxine (T4) when thyrotropin screening results are abnormal. In addition to free T4 immunoassays, the menu of most automated immunoassay instruments includes a nonisotopic version of the original triiodothyronine (T3)-uptake assay called "T-uptake." We evaluated the ability of five such assays (Access, ES-300, IMx, Magnum Opus, and Stratus) to accurately estimate the free thyroxine index (FTI) in euthyroid, hyperthyroid, and hypothyroid patients with abnormal concentrations of thyroid hormone-binding proteins, and in patients with nonthyroidal illness. For comparison, we calculated a similar FTI, using either T3-uptake or direct measurement of thyroxine-binding globulin (TBG). Euthyroid reference ranges were comparable. Of euthyroid patients with increased TBG, 12-32% and 5-20% had increased or suppressed FTI, respectively, depending on the T-uptake method used. Except for IMx, 6-35% of hypothyroid patients with increased TBG had inappropriately increased FTI. Patients with nonthyroidal illness had comparable results regardless of the method used, and T-uptake methods were variably affected by known inhibitors of thyroid hormone binding. The most reliable T-uptake method appeared to be the IMx, which, despite claims that it measures all thyroid hormone-binding proteins, correlated best with TBG concentrations.

Aspirin↗
PubMed1975

The one-step approach to thyroid function tests. The diagnostic thyroxine ratio (DTR)/serum thyroxine (T4) level combination.

A one-step approach to thyroid function tests is reported, using Tetralute kits (Ames). Two parameters are obtained simultaneously--namely, the serum thyroxine (T4) level and the diagnostic thyroxine ratio (DTR), which correlates well with the free thyroxine index (FTI) as determined by two separate tests-the T4 level and the triiodothyronine resin uptake ((T3U)) estimations. Data presented from patients with a variety of thyroid states prove that the DTR, particularly in combination with the T4 level, is adequate for the assessment of thyroid dysfunction in almost all circumstances.

Adolescent↗
PubMed2004

[Bioequivalence of L-thyroxine tablets and a liquid L-thyroxine solution in the treatment of hypothyroid patients].

BACKGROUND AND PURPOSE: The therapeutic equivalence of L-thyroxine (T(4)) tablets and a new liquid T(4) solution was investigated in patients with hypothyroidism. PATIENTS AND METHODS: The study was unblinded, multicentric and prospective: 136 patients (with autoimmune thyroiditis, after thyroid surgery and/or after therapy with radioiodine), who had been treated with T(4) tablets, were switched to a treatment with a liquid T(4) solution (containing 5 microg T(4) per drop) in the same dosage. Thyrotropin (TSH) was measured after 1, 2, 3, and 6 months; the dose of the T4 solution was adapted to obtain a midnormal TSH level. RESULTS: The T(4) dose remained constant during the 6 months. Dose adaptations were necessary in 19-23% of patients at the different timepoints, with dose reductions and increases, respectively, becoming equally frequently necessary. The mean TSH level was 0.79 mU/l at the beginning and 0.81 mU/l after 6 months; similarly, the mean levels of free triiodothyronine (fT(3)) and free thyroxine (fT(4)) were equivalent. The liquid T(4) solution was well tolerated: adverse events were noted in three patients (1.8%); one serious adverse event (thyroid carcinoma) showed no correlation to the treatment. CONCLUSION: T(4) tablets and liquid T(4) solution have been proven to be therapeutically equivalent; the use of liquid T(4) solutions will have advantages in the treatment of hypothyroid infants or elderly patients with disturbed swallowing, patients who need a differentiated titration of the T(4) dose, as well as patients with an allergy against inactive ingredients of L-thyroxine tablets.

Adolescent↗
PubMed1982

Synthesis of thyroid hormone metabolites by photolysis of thyroxine and thyroxine analogs in the near UV.

Photolysis of thyroxine and its analogs in the near UV permitted synthesis in good yield of picogram to gram quantities of thyroid hormone metabolites. Preparation of the same metabolites by classical chemical synthesis requires multistep procedures. Specifically labeled metabolites of high specific activity (e.g., those carrying the label in the nonphenolic ring) were obtained by photolysis of appropriately labeled thyroxine or 3',3',5'-triiodothyronine (reverse triiodothyronine). Some of these labeled metabolites, which are required for metabolic studies (3-iodothyronine and 3,3'-diiodothyronine, labeled in the nonphenolic ring), had not previously been obtained by other methods. Irradiation of thyroxine and reverse triiodothyronine in 150 mM methanolic ammonium hydroxide with greater than 340-nm light caused removal of one iodine atom from the phenolic ring with formation of 3,5,3'-triiodothyronine and 3,3'-diiodothyronine, respectively. Irradiation with higher-energy light (greater than 300 nm) led to stepwise removal of additional iodine atoms. Those in the phenolic ring were removed preferentially, so that 3,5-diiodothyronine and 3-iodothyronine, respectively, were formed. The iodine atoms in the nonphenolic ring were lost more slowly. Tetraiodothyroacetic acid followed a similar photodeiodination pattern. Photolysis with light in the near UV is a simple method for the synthesis of thyroid hormone metabolites.

Diiodothyronines↗
PubMed1995

An enzymeimmunoassay for total thyroxine using avidin-biotin separation system and thyroxine-peroxidase conjugate.

An avidin-biotin enzymeimmunoassay for total thyroxine in serum is described. Avidin was adsorbed to biotinylated bovine serum albumin coated tubes prepared with glutaraldehyde as coupling agent. In the enzymeimmunoassay, affinity purified biotinylated anti-thyroxine IgG, sample or standards, and thyroxine-horseradish peroxidase were simultaneously added to the avidin coated tubes. The bound enzymatic activity was then measured with o-phenylenediamine and H2O2. Results showed that the assay has good precision (within-assay CV% less than 10% and between assay 11.7% in hypo- and 6.9% in hyperthyroid range), good assay range (0-800 nmol/L), good sensitivity (4 nmol/L), and can be performed in 2.5 hours. The results obtained correlate well (r = 0.93) with those of an RIA.

Animals↗
PubMed2003

Thyroxine treatment modified in infertile women according to thyroxine-releasing hormone testing: 5 year follow-up of 283 women referred after exclusion of absolute causes of infertility.

BACKGROUND: Mild hypothyroidism may contribute to disturbed reproductive function. We hypothesized that frequent thyroxine-releasing hormone (TRH) testing to fine-tune thyroxine (T4) therapy instituted upon every TRH-induced thyroid-stimulating hormone (TSH) rise above the mean of a healthy population (i.e. 15 mIU/l) would improve fecundity compared with historical data. METHODS: In a cohort of 283 infertile women followed over 5 years, we assessed (i) pregnancy, abortion and delivery rates, (ii) thyroid function over time in women who conceived compared with those who did not, and (iii) various thyroid parameters with respect to fertility. RESULTS: Overall conception rate of 37% was higher (P < 0.05) than previously reported and independent of thyroid function prior to T4 therapy, thyroxine dose or elevated thyroid autoantibodies. Never achieving basal TSH <2.5 IU/l or TRH-stimulated TSH <20 mIU/l with T4 therapy resulted in lower conception rates (P < 0.05). Median time to conception was 6 months, but 18 months in women who declined TRH testing (P < 0.02). Overall abortion rate was 9%. Only first trimester miscarriages occurred. CONCLUSIONS: Based on the presented protocol, high pregnancy and parturition rates were observed. Whether this is due to early T4 therapy remains to be determined. Abortions appeared to be associated with higher TSH but not with elevated thyroid antibodies.

Abortion, Spontaneous↗
PubMed1981

Specific anti-thyroxine antisera induced by thyroxine sensitized liposomes.

Specific anti-thyroxine rabbit antisera were generated from complete Freund's adjuvant with liposomes consisting of sphingomyelin, cholesterol, dicetylphosphate and 5-N-thyroxine-2,4-dinitrophenyl-phosphatidylethanolamine (T4-Dnp-PE). Spin membrane immunoassay technique was used to measure the sensitivity and specificity of these antisera. Addition of 10 to 70 ng of L-thyroxine produced significant inhibition of immune lysis. Addition of L-3,3',5-triiodothyronine (T3) up to 800 ng showed no cross reaction.

Amines↗
PubMed1987

[Cytoplasmic thyroxine-binding protein: the role of RNA in the interactions between thyroxine receptor and cell nuclei].

The presence of 3H-orotic acid in the cytoplasmic receptor fraction isolated in our laboratory, the sensitivity of this fraction to treatment with RNAases accompanied by a shift of the absorption peak of the receptor preparation towards the long-wave region as well as the use of the absorption filter technique point to the existence of ternary thyroxine-thyroxine-binding protein-RNA complex in the cytoplasm. It was found that the cytoplasmic hormone-receptor complex of thyroxine is a genetically active form which can interact with the nuclei and modify the activity of chromatin. The role of RNA in these interactions is discussed.

Animals↗
PubMed1986

Effects of thyrotropin-releasing hormone on serum concentrations of thyroxine and triiodothyronine in healthy, thyroidectomized, thyroxine-treated, and propylthiouracil-treated dogs.

Effects of thyrotropin-releasing hormone (TRH) on serum concentrations of thyroid hormones were studied in 36 mixed-bred dogs. Dogs were randomly assigned to 7 groups. Significant increases (P less than 0.05) of serum thyroxine (T4) values occurred as early as 2 hours and reached a peak at 6 to 8 hours after IV injection of 300 to 1,100 micrograms of TRH. Thyroxine concentrations in response to a TRH dose greater than 500 micrograms were similar to those observed with the 300-micrograms dose. Transient coughing, vomiting, salivation, and defecation after large doses (900 and 1,100 micrograms) were observed. Mean serum T4 concentration decreased from 2.1 micrograms/dl to 0.9 micrograms/dl within 1 day of thyroidectomy. Clinical signs of hypothyroidism, including lethargy, dry coats, and diffuse alopecia, were present in 2 dogs at a month after surgical operation. Thyroxine concentrations were detectable for greater than 2 months. Injection (IV) of 700 micrograms of TRH 6 weeks after surgical operation had no effect on serum concentration of T4 in thyroidectomized dogs. In 5 T4-treated dogs, TRH (700 micrograms, IV) significantly increased the serum T4 value, indicating that pituitary thyrotropes were responsive to TRH, in spite of daily medication of 0.8 mg of T4. Four dogs were treated orally with 200 mg of propylthiouracil/day for 5 weeks. Intravenous injection of 700 micrograms of TRH in propylthiouracil-treated dogs had no effect on the serum T4 concentration, indicating that TRH had no effect on serum T4 values in these dogs during the experimental period. These results indicate that TRH can replace bovine thyrotropin for the canine thyroid function test.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1985

Estimation of thyroxin, triiodothyronine, thyrotropin, free thyroxin, and triiodothyronine uptake by use of magnetic-particle solid phases.

We evaluated solid-phase radioassays involving small, uniform magnetic particles (MAGIC, Corning Medical), obtaining data on total serum thyroxin (T4), triiodothyronine (T3), thyrotropin (TSH), free thyroxin (FT4), and T3 uptake for a total of 301 serum samples from euthyroid patients; patients receiving replacement thyroxin; patients receiving estrogen, or who were pregnant; hyperthyroid, hypothyroid, and nonthyroidally ill patients; and patients receiving salicylates, phenytoin, or heparin. We found each procedure to be technically simple and precise (between-assay CVs for T4 2.8-8.2%, T3 9-7.4%, TSH 2.9-4.5%, FT4 3.6-11%, T3 uptake 2.2-3.7%). The magnetic separations are rapid (2 to 5 min), reproducible, and complete. Each MAGIC assay produced clinically appropriate results. These assay systems offer the convenience associated with noncentrifugation assays and excellent analytical performance.

Binding Sites↗