Interferences With Thyroid Function Immunoassays: Clinical Implications and Detection Algorithm.

Automated immunoassays used to evaluate thyroid function are vulnerable to different types of interference that can affect clinical decisions. This review provides a detailed overview of the six main types of interference known to affect measurements of thyroid stimulating hormone (TSH), free thyroxine (T4) and free triiodothyronine (T3): macro-TSH, biotin, antistreptavidin antibodies, anti-ruthenium antibodies, thyroid hormone autoantibodies, and heterophilic antibodies. Because the prevalence of some of these conditions has been reported to approach 1% and the frequency of testing for thyroid dysfunction is important, the scale of the problem might be tremendous. Potential interferences in thyroid function testing should always be suspected whenever clinical or biochemical discrepancies arise. Their identification usually relies on additional laboratory tests, including assay method comparison, dilution procedures, blocking reagents studies, and polyethylene glycol precipitation. Based on the pattern of thyroid function test alterations, to screen for the six aforementioned types of interference, we propose a detection algorithm, which should facilitate their identification in clinical practice. The review also evaluates the clinical impact of thyroid interference on immunoassays. On review of reported data from more than 150 patients, we found that ≥50% of documented thyroid interferences led to misdiagnosis and/or inappropriate management, including prescription of an unnecessary treatment (with adverse effects in some situations), inappropriate suppression or modification of an ongoing treatment, or use of unnecessary complementary tests such as an I123 thyroid scan. Strong interaction between the clinician and the laboratory is necessary to avoid such pitfalls.

Thyroid function alterations attributed to high iodide supplementation in maternal rats and their offspring.

OBJECTIVE: Our aim was to investigate thyroid function alterations attributed to high iodide supplementation in maternal rats and their offspring. METHODS: Depending on their iodide intake, the pregnant rats were randomly divided into three groups: normal iodide intake (NI), 10 times high iodide intake (10 HI) and 100 times high iodide intake (100 HI) groups. Iodine concentration in the urine and maternal milk; iodine content and mitochondrial superoxide production; expression of TRα1, TRß1, NIS and Dio1 in both the thyroid and mammary glands were all measured. The offspring were exposed to different iodide-containing water (NI, 10 HI and 100 HI) from weaning to postnatal day 180 (PN180). Serum thyroid hormone levels were measured in both maternal rats and their offspring. RESULTS: Iodine concentration in the urine and maternal milk, as well as iodine content in the thyroid and mammary glands was significantly increased in both the 10 HI and 100 HI groups (p < .05). In the 100 HI group of maternal rats, low FT3 levels, high FT4, TPOAb and TgAb levels were detected. In addition, an increased mitochondrial superoxide production and decreased expression of TRα1, TRß1, NIS and Dio1 in the thyroid and mammary glands was found (p < .05). A positive staining of CD4+ that co-localized with TRß1 in the infiltrated cells within the thyroid follicles was observed. At PN180 in the offspring, the FT3 and FT4 levels showed a significant decrease, while the levels of serum TSH, TPOAb and TgAb were significantly increased in both 10 HI and 100 HI groups (p < .05). CONCLUSION: In maternal rats, although normal thyroid function can be maintained following 10 HI, thyroiditis can be induced following 100 HI on lactation days 7, 14, and 21. In the offspring at PN180, hypothyroidism complicated with thyroiditis can occur in both the 10 HI and 100 HI groups.

Thyroxine and triiodothyronine content in commercially available thyroid health supplements.

BACKGROUND: As defined by the Dietary Supplement Health and Education Act 1997, such substances as herbs and dietary supplements fall under general Food and Drug Administration supervision but have not been closely regulated to date. We examined the thyroid hormone content in readily available dietary health supplements marketed for "thyroid support." METHODS: Ten commercially available thyroid dietary supplements were purchased. Thyroid supplements were dissolved in 10 mL of acetonitrile and water with 0.1% trifloroacetic acid and analyzed using high-performance liquid chromatography for the presence of both thyroxine (T4) and triiodothyronine (T3) using levothyroxine and liothyronine as a positive controls and standards. RESULTS: The amount of T4 and T3 was measured separately for each supplement sample. Nine out of 10 supplements revealed a detectable amount of T3 (1.3-25.4 µg/tablet) and 5 of 10 contained T4 (5.77-22.9 µg/tablet). Taken at the recommended dose, 5 supplements delivered T3 quantities of greater than 10 µg/day, and 4 delivered T4 quantities ranging from 8.57 to 91.6 µg/day. CONCLUSIONS: The majority of dietary thyroid supplements studied contained clinically relevant amounts of T4 and T3, some of which exceeded common treatment doses for hypothyroidism. These amounts of thyroid hormone, found in easily accessible dietary supplements, potentially expose patients to the risk of alterations in thyroid levels even to the point of developing iatrogenic thyrotoxicosis. The current study results emphasize the importance of patient and provider education regarding the use of dietary supplements and highlight the need for greater regulation of these products, which hold potential danger to public health.

Thyroid hormones in milk and blood of lactating donkeys as affected by stage of lactation and dietary supplementation with trace elements.

The traditional utilization of donkeys (Equus asinus) as dairy animals has recently attracted substantial scientific interest with regard to human nutrition. Donkey milk is well tolerated by infants with cows' milk allergy, useful in the treatment of human immune-related diseases, in the prevention of atherosclerosis, and in-vitro studies showed an anti-proliferative effect. Active 3-3'-5-triiodothyronine (T3) in colostrum and milk could play different physiological roles, systemic and paracrine, for both the mother and the suckling offspring. The aim was to evaluate whether thyroid hormones (TH) concentrations in milk and blood of lactating donkeys change with the advancing lactation and whether they can be affected by dietary supplementation with several trace elements, some of them directly involved with TH synthesis (I), metabolism (Se) and action (Zn). Sixteen lactating jennies were divided into two groups (CTL and TE). Mixed feed for TE was added with Zn, Fe, Cu, Mn, I, Se and Co. Every 2 weeks milk and blood samples were collected at 11·00. Total concentrations of T3 in milk (T3M) and T3 and T4 in plasma (T3P and T4P) were assayed using ELISA kits, validated for the donkey species. T3M was not correlated with TH concentrations in blood, did not change with the stage of lactation, and was significantly higher in TE (4·09 ± 0·07 ng/ml, mean ± SE) than in CTL group (3·89 ± 0·08 ng/ml). T4P (81·8 ± 5·2 ng/ml) and T3P (15·2 ± 1 ng/ml) significantly changed with time, but were not significantly affected by dietary treatment. T3P/T4P ratio was significantly lower in TE group. This study indicates that in donkey milk the concentration of T3, a human-like bioactive compound, can be affected by trace elements intake.

Estradiol-17beta and linseed meal interact to alter visceral organ mass and hormone concentrations from ovariectomized ewes.

To evaluate the estrogenic potential of secoisolariciresinol diglycoside (SDG) found in linseed meal (LSM) on visceral organ mass, IGF-I, and thyroid hormone (T(3) and T(4)) concentrations, 48 multiparous, ovariectomized ewes (54.6 +/- 1.1 kg) were used in a 3 x 4 factorial arrangement. Main effects were length of LSM feeding (0, 1, 7, or 14 d) and length of exposure to estradiol-17beta (E(2)) implant (0, 6, or 24 h prior to tissue collection). Implanting ewes with E(2) for 24h increased liver mass relative to empty body weight (EBW; g/kg EBW) compared with ewes implanted for 0 or 6 h (P or= 0.12) CYP2C or CYP3A mRNA expression or cellularity of the liver. Exogenous E(2) influenced circulating concentrations of IGF-I, T(3), and T(4). The estrogenic or anti-estrogenic potential of LSM is dependent upon the tissue, exposure to E(2), and the duration of LSM feeding. Feeding LSM during gestation, lactation, or during the grow-finish phase warrants further investigation.

Tissue-specific thyroid hormone deprivation and excess in monocarboxylate transporter (mct) 8-deficient mice.

Mutations of the X-linked thyroid hormone (TH) transporter (monocarboxylate transporter, MCT8) produce in humans unusual abnormalities of thyroid function characterized by high serum T3 and low T4 and rT3. The mechanism of these changes remains obscure and raises questions regarding the regulation of intracellular availability and metabolism of TH. To study the pathophysiology of MCT8 deficiency, we generated Mct8 knockout mice. Male mice deficient in Mct8 (Mct8(-/y)) replicate the thyroid abnormalities observed in affected men. TH deprivation and replacement with L-T3 showed that suppression of TSH required higher serum levels T3 in Mct8(-/y) than wild-type (WT) littermates, indicating hypothalamus and/or thyrotroph resistance to T3. Furthermore, T4 is required to maintain the high serum T3 level because the latter was not different between the two genotypes during administration of T3. Mct8(-/y) mice have 2.3-fold higher T3 content in liver associated with 6.1- and 3.1-fold increase in deiodinase 1 mRNA and enzymatic activity, respectively. The relative T3 excess in liver of Mct8(-/y) mice produced a decrease in serum cholesterol (79 +/- 18 vs. 137 +/- 38 mg/dl in WT) and an increase in alkaline phosphatase (107 +/- 23 vs. 58 +/- 3 U/liter in WT) levels. In contrast, T3 content in cerebrum was 1.8-fold lower in Mct8(-/y) mice, associated with a 1.6- and 10.6-fold increase in D2 mRNA and enzymatic activity, respectively, as previously observed in TH-deprived WT mice. We conclude that cell-specific differences in intracellular TH content due to differences in contribution of the various TH transporters are responsible for the unusual clinical presentation of this defect, in contrast to TH deficiency.

Thyroid hormones (TH) and 5'-monodeiodinase (5'-MD) activity in goat's milk from the early, mid- and late lactation period.

The physiological significance of thyroid hormones (TH) present in colostrum and milk is still under consideration. The present study was aimed at determining milk thyroxine (T4) and triiodothyronine (T3) levels in three lactation phases (early, mid- and late) of the goat, and to measure activity of the milk 5'-deiodinase (5'-MD) enzyme responsible for the intramammary conversion of prohormone T4 to its metabolically highly active form T3. Thirty-two milk goats (Polish White breed) fed a standard diet were used for milk sampling. The highest TH levels in mammary secretion were recorded during the first 2-3 days post partum. Then the hormone levels decreased, and by about Day 7 fluctuated around the overall mean for the early-lactation phase (Days 1 to 24 of lactation), recording 0.134+/-0.059 microg T4 and 150.8+/-2.80 ng T3 in 100 ml of the milk. Such T4 concentrations appeared to be comparable to those in the rabbit and human, whereas the concentration of T3 was higher than in the cow, pig and mare's milk. Milk 5'-MD activity was higher (P < 0.01) during early and late lactation, compared to the mid-lactation phase. It coincided with low T4 and high T3 milk levels during early lactation, and with high milk T4 and low T3 concentrations during late lactation. The quantity of T4 and T3 available to newborn kids in milk suggests that TH ingested with the colostrum may have a physiological role during the early postnatal life of suckling goats.

The quantity of thyroid hormone in human milk is too low to influence plasma thyroid hormone levels in the very preterm infant.

BACKGROUND: Thyroid hormone is crucial for brain development during foetal and neonatal life. In very preterm infants, transient low levels of plasma T4 and T3 are commonly found, a phenomenon referred to as transient hypothyroxinaemia of prematurity. We investigated whether breast milk is a substantial resource of thyroid hormone for very preterm neonates and can alleviate transient hypothyroxinaemia. Both the influence of breast feeding on plasma thyroid hormone levels and the thyroid hormone concentration in preterm human milk were studied. METHODS: Two groups were formed from the placebo group of a randomized thyroxine supplementation trial in infants born at < 30 weeks' gestational age on the basis of the mean breast milk intake during the third, fourth and fifth weeks of life. One group received more than 50% breast milk (mean breast milk intake 84%, n = 32) and the other group less than 25% breast milk (mean breast milk intake 3.3%, n = 25). Plasma thyroid hormone concentrations were compared between the two groups. Breast milk was collected from mothers of infants participating in the same trial and the thyroxine concentration in breast milk was measured with RIA after extraction. RESULTS: No significant differences were found between both groups in plasma concentrations of T4, free T4, T3, TSH, rT3 and thyroxine-binding globulin (TBG), which were measured once a week. Thyroxine concentration in breast milk ranged between 0.17 microg/l and 1.83 microg/l (mean 0.83, SD 0.3 microg/l) resulting in a maximum T4 supply of 0.3 microg/kg via ingested breast milk. In formula milk, the T4 concentration was equally low. Protease treatment did not influence the measured T4 concentrations. CONCLUSIONS: No differences in plasma thyroid hormone between breast milk-fed and formula-fed infants were found. The amount of T4 present in human milk and formula milk is too low to alter the hypothyroxinaemic state of preterm infants.

Hemodynamic effects of thyroidectomy in sedentary horses.

OBJECTIVE: To investigate hemodynamic effects of thyroidectomy in horses at rest. ANIMALS: 6 healthy aged Quarter Horse mares. PROCEDURE: Horses were monitored for 5 months before and 4 weeks after thyroidectomy and for an additional 4 weeks after administration of thyroid hormone supplement (2.5 microg of thyroxine/kg of body weight, PO, q 12 h, and 0.6 microg of triiodothyronine/kg, PO, q 12 h). Responses to thyroid-stimulating hormone (TSH) were measured before and 4 weeks after thyroidectomy. Other variables monitored daily were resting rectal temperature (T), heart rate (HR), respiratory rate (RR), and body weight (BW). Monthly cardiac output (Q), blood volume (BV), plasma volume (PV), standard electrocardiographic measures, systolic and right ventricular blood pressure, and HR responses were determined after IV administration of isoproterenol and phenylephrine. Variables were analyzed by use of repeated-measures ANOVA. RESULTS: Complete thyroidectomy was confirmed by minimal response to TSH 4 weeks after surgery. Resting HR, RR, T, Q, and beta-adrenergic responsiveness to isoproterenol decreased significantly after thyroidectomy. Resting T, Q, and beta-adrenergic responsiveness increased after administration of supplement and was not significantly different from euthyroid values. Blood volume and PV increased significantly after thyroidectomy but did not return to euthyroid values despite administration of supplement. Response to phenylephrine was minimally different between treatments. CONCLUSIONS AND CLINICAL RELEVANCE: Thyroidectomy in horses caused decreased resting HR, RR, T, Q, and isoproterenol responsiveness and increased BV, PV, PQ interval, and QT interval corrected for HR. Some of these surgically induced changes appeared to be partially reversed by administration of thyroid hormone supplement.

New successful treatment with disinfectant for atopic dermatitis.

For the treatment of atopic dermatitis, a variety of therapies are used including folk medicine. At present, there is no single treatment which is effective to cure the symptoms of atopic dermatitis completely in all patients. We are drawing attention to the high isolation rate of Staphylococcus aureus when starting disinfectant treatment combined with topical steroid therapies for the purpose of killing S. aureus. As a result, we examined many patients in whom almost a complete remission was obtained even after short periods of therapy, though it had been difficult to obtain improvement by conventional treatments. In many patients, IgE values and reagin antibody titer decrease dramatically soon after starting treatment. As a disinfectant, 10% povidone-iodine solution was used. We investigated also the effect of iodine contained in the povidone-iodine solution on the thyroid gland.

The effects of modifying proficiency testing materials on thyroid function test results. A College of American Pathologists Ligand Assay Survey Study.

OBJECTIVE: To gain insight on the matrix effects, and possible clinical implications, resulting from diluting and concentrating proficiency testing survey material used for the measurement of thyroid function tests. DESIGN: To the standard set of five proficiency survey samples, three supplementary "Wildcard" samples were added. These additional samples were manufactured by overfilling and underfilling vials prior to lyophilization so as to vary the thyroxine-binding protein concentrations. Survey participants measured thyroxine, free thyroxine, and the triiodothyronine uptake and related tests on the Wildcard samples. In addition, free thyroxine indices were calculated. SETTING: The first mailing of the 1995 College of American Pathologists (CAP) Ligand Assay--Series 1 Survey. MAIN OUTCOME MEASURES: Results obtained from the regular set of survey samples and the Wildcard set were compared to values expected by the laws of conservation of matter and mass action. PARTICIPANTS: The approximately 2000 participants of the first mailing of the 1995 CAP Ligand Assay--Series 1 Survey. RESULTS: Numerous assays systems did not give the predicted results, including all of the single-step radioimmunoassays for free thyroxine and over three quarters of free thyroxine index determinations. CONCLUSIONS: Varying the dilution of proficiency survey material produced results that were not predicted by the laws of conservation of matter and of mass action. Although these observations may have been the result of matrix effects, one cannot rule out the possibility that certain thyroid assays may not work in clinical situations having abnormal thyroxine-binding protein concentrations.

Selenium deficiency alters thyroid hormone metabolism in guinea pigs.

In guinea pigs, activity of glutathione peroxidase in most organs is markedly lower than in organs of other rodents despite comparable dietary intakes and tissue levels of selenium. To determine if metabolism of selenium with respect to other selenoproteins also differs in guinea pigs, we measured the effects of selenium intake on thyroid hormone metabolism. Weanling male Hartley Albino guinea pigs were fed a selenium-deficient Torula yeast-based diet, or the same diet supplemented with 0.5 mg selenium/kg diet as sodium selenate for 72 d. Growth was impaired in guinea pigs fed the unsupplemented diet. Activity of glutathione peroxidase was higher in tissues and plasma of supplemented guinea pigs than in selenium-deficient animals. However, it was still far lower than reported values for other rodent species. In selenium deficiency, activity of type 1 5'-iodothyronine deiodinase was 60% less in liver and 45% less in kidney. Concentration of thyroxine was 68% lower in kidney of selenium-deficient animals, and levels of 3,3',5-triiodothyronine in kidney and plasma were 44 and 31% lower, respectively. Thus, with the exception of thyroxine concentrations, thyroid hormone metabolism responds to selenium deficiency in guinea pigs as it does in rats, although the magnitude of that response is not as great.

Modulation of fetal and placental metabolic pathways in response to maternal thyroid and glucocorticoid hormone excess.

Triiodothyronine (T3) treatment of pregnant rats for 6 days, 10 micrograms/100 g, resulted in a pronounced induction of enzymes related to gluconeogenesis and lipogenesis and of mitochondrial FAD-glycerophosphate dehydrogenase in the maternal liver, as previously observed in male rats. There was virtually no change in the activity of these enzymes in the placenta. However, there was a distinct induction of these enzymes in the fetal liver, even if increments in fetal serum and liver T3 were much smaller than on the maternal side. This indicates that changes in hepatic enzyme activities are a more sensitive index of fetal hyperthyroidism than T3 levels. The increased lipogenic capacity was expressed by greater incorporation of a tritium tracer into fatty acids. Administration of triamcinolone, 2 mg/100 g, for the last 5 days of gestation resulted in marked induction of maternal hepatic enzymes of lipogenesis, gluconeogenesis and of aspartate aminotransferase (ASAT), known to occur in male rats, as well as in a metabolic pattern of insulin resistance. The response of placental enzymes was limited to a small elevation in ASAT and phosphoenolpyruvate carboxykinase (PEPCK) activity. In the fetal liver there was no stimulation of lipogenic enzymes, but a marked induction of PEPCK and ASAT. The changes in the lipogenic capacity were confirmed by tritium incorporation into serum and liver fatty acids. These results demonstrate the marked sensitivity of specific fetal enzyme systems to the maternal iatrogenic hyperthyroidism or hypercorticism. The limited alterations in placental enzyme activities are in accord with the concept that placental metabolic stability fulfils a protective function toward the fetus.

[L-thyroxine and triiodo-L-thyronine in the cerebrospinal fluid and hypothalamo-hypophyseal axis of hyper- or hypothyroid rats]. / L-tiroxina y triiodo-L-tironina en líquido cefalorraquídeo y eje hipotálamo hipofisario de ratas, hiper- e hipotiroideas.

L-thyroxine and triiodo-L-thyronine concentrations in cerebrospinal fluid (CSF), hypothalamus and pituitary gland are measured in male albino-Wistar rats under several experimental thyroid disfunction : including hyperthyroidism induced by L-T3 and L-T4 treatments and surgical hypothyroidism. Radioimmunoassay is carried out by Nejad's method modified in this work. The pattern of thyroid hormone concentrations in CSF is similar to that in serum, but the values obtained are lower. Thyroid hormone concentrations in adenohypophysis as opposed to hypothalamus or cerebral cortex, show an inverse change to functional thyroid status.

[Thyroid biological evaluation: critical study]. / Le bilan biologique thyroïdien: étude critique.

Laboratory tests to assess thyroid gland function have changed considerably over the past few years, with the development of techniques allowing for the direct routine determination of unbound thyroid hormones (tri and tetra-iodothyronine), the "ultra-sensitive" assay of serum concentrations of TSH (thyroid-stimulating hormone) and finally, the radioimmunological assay of anti TSH-receptor antibodies. In our study, which excluded patients having an excess of iodine or who had serious disease of an organ other than the thyroid, we assessed the impact of these parameters respectively on various disease categories. An innovation has been made in that anti TSH-receptor antibody assay is now possible in everyday practice while up to the present this was only possible in the hospital setting and with a limited number of cases: this titer is important as a classification parameter in diagnosing Grave's disease and has prognostic importance to monitor treatment of patients suffering from this disorder. The "ultrasensitive" version of measuring thyrotropic hormone requires a new strategy: TSH assessment becomes the first-line diagnosis test to evaluate thyroid function because it differentiates from control subjects, as well as patients with hyper ou hypothyroidism who are not receiving therapy. In other cases, dosing of the free hormones T3 and T4 remains a vital supplementary test.

[Role of breast feeding in providing the newborn infant with thyroid hormones]. / Rol' estestvennogo vskarmlivaniia v obespechenii organizma novorozhdennogo gormonami shchitovidnoi zhelezy.

Radioimmunoassay was conducted to study the content of triiodothyronine and thyroxine in the human milk and in the blood serum of mature and premature newborns. It was established that with the growth of the lactation period the content of triiodothyronine and thyroxine rose; it was, respectively, 0.5 +/- 0.12 and 100.3 +/- 8.4 nmol/l in colostrum, 9.2 +/- 0.7 and 271.3 +/- 14.1 nmol/l in transient milk, 11.1 +/- 1.07 and 405.0 +/- 21.4 nmol/l in fully formed milk. Fully formed milk contains three times more triiodothyronine and thyroxine than the blood serum of newborns. The mature newborns have a sufficiently formed thyroid function. The level of thyroid hormones in the blood serum of premature newborns is decreased in proportion to the stage of their prematurity. They receive the deficient hormones through the human milk. The authors recommend that the nutrition of premature newborns receiving mixed and artificial feeding should be supplemented with thyroid hormones.

Effects of maternal iodine deficiency on the L-thyroxine and 3,5,3'-triiodo-L-thyronine contents of rat embryonic tissues before and after onset of fetal thyroid function.

Female rats were placed on a low iodine diet (LID) or LID supplemented with KI. They were mated 3-6 months later. Maternal and embryonic tissues were obtained both before the onset of fetal thyroid function, at 11 and 17 days of pregnancy, and at 21 days of gestation. T4 and T3 concentrations were measured by RIA. T4 concentrations were very low in the plasma, liver, and lung of LID dams and in all embryonic samples obtained from such mothers, namely 11-day-old embryotrophoblasts, 17-day-old placentas and embryos, 21-day-old placentas, embryos, plasma, liver, lung, and carcass (whole embryos minus the trachea, thyroid, blood, liver, and brain). T3 was low in 17-day-old placentas and embryos and in all fetal tissues obtained at 21 days of gestation from LID dams. These results show that when iodine deficiency is severe enough to result in very low maternal plasma T4 levels, embryonic tissues are deficient in T4 and T3 both before and after the onset of fetal thyroid function. This finding might be relevant to the etiopathology of human iodine deficiency disorders.

Tri-iodothyronine and thyroxine in human milk.

The concentration of tri-iodothyronine (T3) and thyroxine (T4) in human milk was determined by radioimmunoassay (RIA). The analysis of T3 was performed on unextracted milk and on ethanol extracts of defatted milk. Analysis of unextracted milk was complicated by artifacts. Reliable and reproducible results were achieved only with the milk extracts. In 10 colostral milk samples the mean T3 levels +/- SD were 0.80 +/- 0.52 nmol/l before feeding (early milk) and 0.93 +/- 0.62 after feeding (hind milk). The T3 concentration in colostrum did not change significantly during the feeding to the infant. In 12 mature milk samples collected between infant feedings, the mean T3 +/- SD was 1.19 +/- 0.42 nmol/l. T4 was not detected in any of the samples analysed (detection limit 3 nmol/l).