Effects of electromagnetic fields on bone loss in hyperthyroidism rat model.

Optimal therapeutics for hyperthyroidism-induced osteoporosis are still lacking. As a noninvasive treatment, electromagnetic fields (EMF) have been proven to be effective for treating osteoporosis in non-hyperthyroidism conditions. We herein systematically evaluated the reduced effects of EMF on osteoporosis in a hyperthyroidism rat model. With the use of Helmholtz coils and an EMF stimulator, 15 Hz/1 mT EMF was generated. Forty-eight 5-month-old male Sprague-Dawley rats were randomly divided into four different groups: control, levothyroxine treated (L-T4), EMF exposure + levothyroxine (EMF + L-T4), and EMF exposure without levothyroxine administration (EMF). All rats were treated with L-T4 (100 mg/day) except those in control and EMF groups. After 12 weeks, the results obtained from bone mineral density analyses and bone mechanical measurements showed significant differences between L-T4 and EMF + L-T4 groups. Micro CT and bone histomorphometric analyses indicated that trabecular bone mass and architecture in distal femur and proximal tibia were augmented and restored partially in EMF + L-T4 group. In addition, bone thyroid hormone receptors (THR) expression of hyperthyroidism rats was attenuated in EMF + L-T4 group, compared to control group, which was not observed in L-T4 group. According to these results, we concluded that 15 Hz/1 mT EMF significantly inhibited bone loss and micro architecture deterioration in hyperthyroidism rats, which might occur due to reduced THR expression caused by EMF exposure. Bioelectromagnetics. 38:137-150, 2017. © 2016 Wiley Periodicals, Inc.

Treatment of Hyperthyroidism Reduces Systemic Oxidative Stress, as Measured by Markers of RNA and DNA Damage.

BACKGROUND: Whole-body oxidative stress can be estimated by the urine excretion of oxidized guanosine species, 8-oxo-7,8-dihydroguanosine (8-oxoGuo) and 8-oxo-7,8-dihydro-2'-deoxyguanosine (8-oxodG), derived from RNA and DNA, respectively. These oxidative stress markers are not well explored in thyroid disorders. OBJECTIVE: We aimed to determine whether treatment of hyperthyroid patients affects the levels of these oxidative stress markers. METHODS: Urinary excretion of 8-oxoGuo and 8-oxodG was measured in 51 hyperthyroid patients (toxic nodular goiter [TNG], n = 30; Graves disease [GD], n = 21) before or shortly after initiation of therapy and when stable euthyroidism had been achieved for at least 12 months. RESULTS: Adjusting for age, the baseline urinary excretion of oxidative stress markers correlated positively with plasma thyroxine (8-oxoGuo, P = 0.002; 8-oxodG, P = 0.021) and was significantly higher in GD than in TNG patients (P = 0.001 for both oxidative stress markers). Restoration of euthyroidism significantly affected the excretion of the oxidative stress markers. In TNG, 8-oxoGuo decreased from geometric mean 2.11 nmol/mmol creatinine (95% CI, 1.85-2.39) to 1.91 nmol/mmol (95% CI, 1.67-2.19; P = 0.001), while 8-oxodG decreased from 1.65 nmol/mmol (95% CI, 1.41-1.93) to 1.48 nmol/mmol (95% CI, 1.27-1.74; P = 0.026). In GD, 8-oxoGuo decreased from 2.25 nmol/mmol (95% CI, 1.95-2.59) to 1.79 nmol/mmol (95% CI, 1.63-1.97; P = 0.0003), while 8-oxodG decreased from 2.02 nmol/mmol (95% CI, 1.73-2.38) to 1.54 nmol/mmol (95% CI, 1.31-1.81; P = 0.001). In the euthyroid state, there were no differences between groups. CONCLUSION: Restoration of euthyroidism in patients with hyperthyroidism significantly decreased the systemic oxidative stress load by 10% to 25%. Our findings may help to explain the higher morbidity and mortality linked to hyperthyroid diseases, as shown in observational studies.

Retinol-binding protein in serum and urine of hyperthyroid cats before and after treatment with radioiodine.

BACKGROUND: Retinol-binding protein (RBP) is suggested as a clinically useful marker of renal function in cats. HYPOTHESIS: Serum and urinary RBP concentrations in hyperthyroid (HT) cats differ from those in healthy (H) cats; radioiodine ((131)I) treatment influences serum and urinary RBP concentrations in HT cats. ANIMALS: Ten HT and 8 H cats. METHODS: RBP concentration was evaluated in feline serum and urine samples from a prospective study. RESULTS: There was a significant (P= .003) difference in the urinary RBP/creatinine (uRBP/c) ratios of H (-) and untreated HT (1.4 + or - 1.5 x 10(-2) microg/mg) cats. Serum total thyroxine concentration (1.8 + or - 1.9 microg/dL, 24 weeks) and uRBP/c (0.6 + or - 1.0 x 10(-2) microg/mg, 24 weeks) decreased significantly (P < .001) in HT cats at all time points after treatment with (131)I, and these variables were significantly correlated with one another (r= 0.42, P= .007). Serum RBP concentrations from HT cats (199 + or - 86 microg/L) did not differ significantly (P= .98) from those of H cats (174 + or - 60) and did not change after treatment with (131)I (182 + or - 124 microg/L, P= .80). CONCLUSION AND CLINICAL IMPORTANCE: The presence of urinary RBP in HT cats is a potential marker of tubular dysfunction that is correlated to thyroid status, although it is independent of circulating RBP concentrations. The decreased uRBP/c combined with the absence of changes in serum RBP after treatment suggests that the suspected tubular dysfunction was partly reversible with treatment of (131)I.

Urinary iodide concentration in hyperthyroid cats.

OBJECTIVE: To compare concentrations of urinary iodide (UI) in euthyroid and untreated hyperthyroid cats. ANIMALS: 118 euthyroid and 88 hyperthyroid client-owned cats from 2 nonreferral veterinary practices. PROCEDURES: Iodide concentration was measured in 5 urine samples collected every 3 to 12 months from selected cats, and variability of results between euthyroid cats and hyperthyroid cats prior to the diagnosis of hyperthyroidism was evaluated via 1-way ANOVA, after logarithmic transformation of UI concentrations (logUIs). The UI concentration in hyperthyroid cats was measured at diagnosis and 2 to 6 weeks and 3 to 6 months after treatment for hyperthyroidism. The pretreatment logUI in hyperthyroid cats was compared with that in euthyroid cats, taking into account the effects of renal function on UI concentration. Iodine intake was estimated in euthyroid cats following calculation of the volume of daily urine output, with a fixed value for iodine concentration in feces. RESULTS: The variability of UI concentrations did not differ significantly between hyperthyroid (n = 10) and euthyroid (8) cats. The logUI increased 2 to 6 weeks after initiation of treatment in hyperthyroid cats (n = 80) and was lower in azotemic versus nonazotemic cats. Hyperthyroid cats had a lower logUI than euthyroid cats, and there was no evidence of deficient iodine intake in euthyroid cats. CONCLUSIONS AND CLINICAL RELEVANCE: The logUI was lower in cats with azotemia and with untreated hyperthyroidism, compared with that in euthyroid cats from the same population. Additional studies are needed to determine whether iodine intake plays a role in the development of hyperthyroidism in cats.

Immunoassay of urinary retinol binding protein as a putative renal marker in cats.

The presence of low molecular weight retinol binding protein (RBP) in urine reflects tubular damage. Therefore, RBP has been used as a renal marker in humans and dogs. Using an anti-human RBP antibody (Ab), this study first demonstrates feline urinary RBP by Western blot analysis and then evaluates its potential as a renal marker in cats by enzyme-linked immunosorbent assay (ELISA). Urine was taken by cystocentesis, centrifuged and stored at -80 degrees C until analysis. Urinary RBP levels were compared in clinically healthy cats (H), chronic renal failure patients (CRF) and cats with hyperthyroidism (HT). The detection of a band at the same position as the human RBP standard with Western blot analysis, indicated that RBP was present in the urine of CRF and HT patients but minimally present in H cats. The data obtained with ELISA were in accordance with these observations. RBP levels were expressed as RBP:creatinine (RBP:c) ratios following normalisation with urinary creatinine. The functional assay sensitivity was 1.37 microg/l RBP. Parallelism between the trend lines of the human RBP standard curve and the curves obtained from sequentially diluted urine samples indicated that feline RBP was recovered. The mean intra-assay coefficient of variance was 7% and the standardised agreement index revealed satisfactory day-to-day repeatability. The RBP:c ratio in all H cats (n=10) was below the assay sensitivity. The groups of CRF and HT patients had increased mean RBP:c ratios of 1.6+/-0.5x10(-2) microg/mg (mean+/-SEM, n=10) and 1.4+/-0.4x10(-2) microg/mg (n=13), respectively. Both groups showed a large variation in the relative RBP concentrations of individual cats. In conclusion, RBP is demonstrated for the first time in urine from most CRF and HT patients and the validated ELISA allows its evaluation as a putative renal marker in cats.

Increased urinary excretion of acidic mucopolysaccharides in exophthalmos.

The excretion of mucopolysaccharides normally found in urine (chondroitin, chondroitin sulfates A and C, keratosulfate, and heparitin sulfate) is increased approximately twofold in patients with progresive exophthalmos. A threefold elevation of total serum mucopolysaccharides is also found. These increases are unrelated to thyroid function.

Population living in the Red Sea State of Sudan may need urgent intervention to correct the excess dietary iodine intake.

BACKGROUND: Both inadequate and high intakes of iodine are associated with thyroid disease and associated abnormalities. Consumption of foods deficient in iodine induces hypothyroidism. Conversely, excessive intake of the nutrient precipitates hyperthyroidism. Iodine deficiency causes impairment of thyroid hormonogenesis resulting in goiter (struma), cretinism which is associated with increased prenatal and infant mortality, deafness, motor disabilities and mental retardation due to damage during fetal and neonatal brain development. We have assessed the iodine status of school children from the locality of Port Sudan, Red Sea State of Eastern Sudan. The primary sources of iodine of the children are mainly iodized salt and rations supplied by local donors and various aid agencies operating in the Sudan. METHODS: Male and female children (n=141), aged 6 to 12 years (median age 9.8 years), were selected for the survey using a multistage random sampling technique, between May 22 and August 25, 2006. All the children were assessed for urinary iodine and visible goiter. In addition, the iodine content of twenty salt samples was determined using the lodometric titration method and spot test kits. The components of other foods that are routinely consumed by the children and households were noted using a questionnaire form. FINDINGS: Urinary iodine concentration exceeded 300 microg/l and 1000 microg/l in 65% and 9.9% of the children, respectively. The highest urinary iodine level was 1470 microg/l. The prevalence of visible goiter was 17%. All the salt samples collected from the schools had more than 150mg potassium iodate per kg of salt. CONCLUSIONS: The results of this pilot survey reveal that excessive intake of iodine in children exists in Port Sudan. Inappropriate and unregulated local fortification of salt and lack of monitoring of the imported and donated salt is the primary reason for the excessive intake. There is an urgent need for a regulatory mechanism during the process of iodine fortification and at the point of entry of imported and donated iodized salt as well as the mode of delivery in order to avoid hyperthyroidism and associated disorders. In addition, independent professionals should critically evaluate the health impact of excessive consumption of the nutrient.

Definition of reference ranges for free T4, TSH, and thyroglobulin levels in healthy subjects of the Jaén Health District. / Definición de los rangos de referencia de T4 libre, TSH y tiroglobulina en sujetos sanos del Distrito Sanitario de Jaén.

BACKGROUND AND OBJECTIVES: The treatment guidelines for thyroid dysfunction recommend defining reference ranges for thyroid hormones in each area through assessment of local population data considering the iodine nutritional status. The aim of this study was to define the reference ranges of free thyroxine (FT4), TSH, and thyroglobulin levels in a general population from Jaen, an area of southern Spain with an adequate iodine nutritional status, and whether they were associated with urinary iodine levels. PATIENTS AND METHODS: A cross-sectional study was conducted in 1,003 subjects of the general population of the Jaen Health District. Levels of urinary iodine, FT4, TSH, thyroglobulin, and thyroid peroxidase (TPO) antibodies were measured according to age and sex. RESULTS: Median and mean urinary iodine levels were 110.59µg/L and 130.11µg/L respectively. Median TSH level was 1.83µIU/mL (p2.5=0.56µIU/mL, p97.5=4.66µIU/mL). Median FT4 level was 0.84ng/dL (p2.5=0.62ng/dL, p97.5=1.18ng/dL). TPO antibodies were detected in 5.7% of subjects. There was no correlation between urinary iodine levels and FT4, TSH or TPO antibodies. Subjects with positive TPO antibodies had higher TSH levels (3.34µIU/L versus 2.14µIU/mL, P=.001; odds ratio=2.42). CONCLUSIONS: Urinary iodine levels in Jaen are optimal according to World Health Organization standards. Reference ranges of FT4, TSH, and thyroglobulin do not differ from those reported in the literature and are no associated to urinary iodine levels. The prevalence of positive TPO antibodies was similar to that reported in other Spanish areas.

Urinary iodine concentration of pregnant women and female adolescents as an indicator of excessive iodine intake in Sri Lanka.

BACKGROUND: Mild deficiencies and excesses of iodine have deleterious effects in both females and males. The iodine status of the population after implementation of the universal salt iodization program in Sri Lanka is not known. OBJECTIVE: This cross-sectional study was carried out to assess the iodine status of pregnant women and female adolescents, with urinary iodine concentration used as the measure of outcome. METHODS: The participants were 100 women in the first trimester of pregnancy and 99 female adolescents in Kuliyapitiya, Kurunegala District, North-Western Province, Sri Lanka. The urinary iodine concentration was measured in a casual urine sample from each subject. The iodate contents of salt samples collected from households of the adolescents participating in the study were also measured. RESULTS: The median urinary iodine concentration of 185.0 microg/L and the prevalence of values under 50 microg/L of only 1% among the pregnant women indicate adequate iodine intake and optimal iodine nutrition. The median urinary iodine concentration (213.1 microg/L) among female adolescents indicates a more than adequate iodine intake and a risk of iodine-induced hyperthyroidism. Approximately 8% and 4% of the adolescents and pregnant women, respectively, had urinary iodine concentrations in the range of mild iodine deficiency (51 to 100 microg/L). More than half of the adolescents (56%) and 39% of the pregnant women had urinary iodine concentrations higher than optimal. The median iodine content in salt samples was 12.7 ppm. Only 20.2% of the samples were adequately iodized, and 10.1% of the samples had very high iodine levels. CONCLUSIONS: Female adolescents and pregnant women had no iodine deficiency, but a considerable proportion of them, especially female adolescents, were at risk for iodine-induced hyperthyroidism. There is thus a need for proper monitoring of the salt iodization program to achieve acceptable iodine status.

[Prevalence of autoimmunological disorders of the thyroid in children in the Bialystok population]. / Ocena wysteipowania zaburzen autoimmunologicznych tarczycy u dzieci w populacji bialegostoku.

Up-to-date observations in the regions where effective food iodine supplementation was introduced confirm a significant decrease in goiter incidence. However, in some regions the incidence of goiter remains unchanged or even the increase in the frequency of autoimmunological diseases of the thyroid like, Grave's-Basedov disease and chronic/subacute lymphocytic thyroiditis is observed. The purpose of the study was to evaluate the prevalence of autoimmunological diseases of the thyroid in children aged 6-13 years in the Polish population after 5 years of obligatory iodine supplementation program. The study included 480 school children from 4 elementary schools chosen randomly in the city of Bialystok and 120 patients at the same age treated due to goiter with KI and/or thyroxin for minimum 12 months in the Regional Specialist Outpatient Clinic of Endocrinology. All children underwent physical examination with palpation of goiter and USG of the thyroid. Iodine concentration was assessed in the morning urine by the catalytic method of Sandell-Költhoff. The concentrations ofTSH and antibodies against TSH, thyroid-peroxidase (TPO) and thyroglobulin (Tg) were also determined. The mean concentrations of anti-Tg antibodies were statistically significantly higher in children with goiter in comparison with children with the thyroid gland within the norm (155.8 IU/ml vs. 24.4 IU/ml, p = 0.045). In children with goiter the incidence of'positive' titre of anti-Tg antibodies was 4-fold as high as in children without goiter (33.3% vs. 8.7%, p = 0.0147). High incidence of antibodies against thyroglobulin in the population of children with goiter and a positive correlation between the titre of anti-Tg antibodies and the size of the thyroid in this group suggest a significant role of autoimmunological disorders in the pathogenesis of goiter in the studied population.

Serum and urinary cystatin C in cats with feline immunodeficiency virus infection and cats with hyperthyroidism.

OBJECTIVES: The objective of this study was to investigate serum cystatin C (sCysC) and urinary cystatin C (uCysC) in cats with hyperthyroidism and cats with feline immunodeficiency virus (FIV). METHODS: Thirty cats with FIV, 26 hyperthyroid cats and 28 healthy cats were included. sCysC and uCysC:creatinine (uCysC/uCr) ratio were measured with a human particle-enhanced nephelometric immunoassay, previously validated for feline CysC measurement. Routine renal variables (serum creatinine [sCr], urine specific gravity, urinary protein:creatinine ratio [UPC]) were also measured in the three groups. RESULTS: Cats with hyperthyroidism had significantly higher sCysC and higher uCysC/uCr ratio, lower sCr and a higher UPC than healthy cats. Cats with FIV infection did not show a significantly higher sCysC concentration but had a significantly higher sCr and UPC than healthy cats. uCysC could be detected in only four of them. CONCLUSIONS AND RELEVANCE: This study demonstrated that sCysC is increased in cats with hyperthyroidism, in contrast with sCr, but not in cats with FIV. Many hyperthyroid cats, but only four cats with FIV, had an elevated uCysC/uCr ratio. Further studies may reveal if uCysC might be a valuable marker for tubular dysfunction in cats.

[Urinary cotinine in patients with hyperthyroidism]. / Kotynina w moczu u pacjentów z nadczynnoscia tarczycy.

The aim of our study was an analysis of cotynine, the main nicotine metabolite in the urine among hyperthyroid patients. The study group included 39 females and 4 males. The mean age was 35.59+/-14.22 yrs. (range: 18-73 yrs.; median: 32 yrs) among hyperthyroid patients suffering from: Graves-Basedow disease (GB), Graves' Ophtalmopathy (GO) and toxic nodular goiter (TNG). To evaluate the nicotine smoking intensity and ETS Environmental Tobacco Smoke, the urine analysis of cotynine level were performed. According to the statistical analysis using the Mann-Whitney test, the statistically significant difference between the level of cotynine among smokers suffering from GO and Graves-Basedow disease was revealed (p = 0.03). Similar results were obtained among the GO and TNG (p=0.02) using t-Student test with Welsch correction. To compare, there was no stastistically significant difference between the GB and TNG series (p=0.4). In the group of smoking patients with GO we found out incresed level of urine cotinine than in smoking patients with GB and TNG. We didn't found differences between GB and TNG in depends on an urine analysis of cotynine level.

[Disturbances of thyroid function in adult population of the city of Cracow followed up for ten years observation]. / Zaburzenia funkcji tarczycy w populacji doroslych mieszkanców Krakowa w okresie 10-letniej obserwacji.

UNLABELLED: The aim of the study was to define disturbances of thyroid function in adult population of the city of Cracow followed up for ten years. The analysis included the results obtained from 891 individuals, 571 females and 320 males aged 18-78 years examined prior to and after implementation of the obligatory model of iodine prophylaxis (years 1989-1990 and 1998-1999). After the exclusion of patients diagnosed earlier as having hyperthyroidism or hypothyroidism, the mean TSH level in the years 1998-1999 was significantly higher as compared to data obtained between 1989 and 1990 (1.44 microj/ml vs. 1.30 microj/ml) in the examined population. Our 10 years observation revealed an insignificant increase in frequency of hyperthyroidism only among females (1.6% vs. 0.9%) which did not indicate a clear, endemic in character increase at the population level. In the present investigations, hypothyroidism was demonstrated to occur more frequently as compared to the 1989-90 study (2.1% vs. 1.4% among females and 0.3% vs. 0% in males). Both differences were not statistically significant. As seen from the present results, over the investigated 10-year period, in the Cracow population, there occurred a clear, statistically significant (p<0.001) increase of the percentage of individuals with an elevated TPO antibody titter (3.8% vs. 11.8%). In our investigation no correlation was observed between anti-TPO antibodies and ioduria levels. CONCLUSIONS: The results reveal no statistically significant increase in incidence of hyperthyroidism and hypothyroidism after implementation of the obligatory model of iodine prophylaxis. The future studies are needed to clarify the mechanisms involved in increase of anti-TPO autoantibodies and verify its possible temporary nature.

Longitudinal changes of bone density and bone resorption in hyperthyroid girls during treatment.

Low bone mineral density (BMD) and increased bone turnover are common features of untreated hyperthyroidism in adult patients. The effect of treatment on BMD is still controversial. BMD and bone metabolism in hyperthyroid children have not been thoroughly investigated. In the present study, we measured spinal and whole body BMD by dual-energy X-ray absorptiometry in a group of 13 girls (aged 5.0-14.9 years) at diagnosis of hyperthyroidism. The bone resorption rate was assessed by urine measurement of N-terminal telopeptide of type I collagen (NTX). Hyperthyroid patients have been studied longitudinally during treatment. BMD values and NTX urine concentrations have been also determined in 155 healthy Caucasian girls (aged 2.4-24.2 years). Spinal and whole body bone density measurements were significantly lower compared with healthy controls in untreated hyperthyroid girls, after correction for differences in age and anthropometric measurements (p

Urinary cyclic nucleotide levels in patients with hyper- and hypothyroidism.

Urinary cyclic AMP, cyclic GMP and creatinine excretion were measured in 38 patients with hyperthyroidism, 32 patients with hypothyroidism and in 57 normal subjects. The excretion of both cyclic nucleotides was significantly increased in hyperthyroid females, but not in hyperthyroid males. The cyclic nucleotides/creatinine ratios, however, were uniformly elevated in both male and female hyperthyroid subjects and this was due, in part, to decreased creatinine excretion. Cyclic AMP excretion was significantly decreased in the hypothyroid subjects, but the cyclic AMP/creatinine ratios were not significantly different from normal. There were no significant alterations in cyclic GMP and cyclic GMP/creatinine ratios in the male hypothyroid patients, but ratios in the female patients were slightly greater than in the normals. These results demonstrate that hyper- and hypothyroidism may be associated with appreciable alterations in urinary cyclic nucleotide levels and that there may be sex-related differences in the patterns of urinary excretion of these nucleotides. The cyclic nucleotide levels herein described in patients with hyper- and hypothyroidism are qualitatively and, in some instances, quantitatively similar to those found in patients with hyper- and hypoparathyroidism, respectively.

Urinary excretion of hydroxylysyl glycosides and thyroid function.

Urinary excretion of hydroxylysyl glycosides, two specific collagen metabolites, was measured in 18 patients with hyperthyroidism and 4 patients with hypothyroidism. As in the case of hydroxyproline, values were high in thyrotoxicosis and low in hypothyroidism. The glucosyl-galactosyl-hydroxylysine/galactosyl-hydroxylysine urinary ratio which indicates the bone or skin origin of degraded collagen was found to be unchanged in hyperthyroidism, except in two cases complicated with hypercalcemia where it was very low. This finding provides a further argument in favour of the bone origin of hypercalcemia in thyrotoxicosis.

Urinary excretion of free and conjugated 3',5'-diiodothyronine and 3,3'-diiodothyronine.

RIAs for the estimation of 3',5'-diiodothyronine (3',5'-T2) and 3,3'-diiodothyronine (3,3'-T2) in human urine have been established. The urinary excretion of both glucuronide and sulfate conjugates of T2 and of T4, T3, and rT3 were estimated by means of enzymatic deconjugation. In healthy controls, the mean excretion (picomoles per 24 h) of free T4 was 1820, that of free T3 was 813, that of free rT3 was 77, that of free 3',5'-T2 was 13, and that of free 3,3'-T2 was 674. The total excretion of free and conjugated T4 was 2941, that of T3 was 1283, that of rT3 was 791, that of 3',5'-T2 was 709, and that of 3,3'-T2 was 2688. Significant amounts of sulfated T4 and T3 could not be demonstrated, amounts of sulfated T4 and T3 could not be demonstrated, whereas the excretion of sulfated rT3 was higher than that of glucuronidated rT3 (P less than 0.001). In contrast, glucuronidated and sulfated 3',5'-T2 as well as glucuronidated and sulfated 3,3'-T2 were found in the urine in equal amounts. In hyperthyroidism, the excretions of free and glucuronidated iodothyronines were increased, whereas the increase of the excretions of sulfated iodothyronines were less pronounced, only reaching statistical significance for 3,3'-T2 (P less than 0.02). In hypothyroidism, the excretions of both free, glucuronidated and sulfated iodothyronines were reduced. Significant amounts of sulfated T4 and T3 could not be demonstrated in urine from hyperthyroid or hypothyroid patients. Our data demonstrate that the amounts of free iodothyronines excreted in the urine vary considerably, suggesting active renal handling. The amounts of urinary glucuronidated and sulfated conjugates of the different iodothyronines studied vary considerably and are affected by thyroid function.

A radioimmunoassay for measurement of thyronine and its acetic acid analog in urine.

A sensitive and specific RIA has been developed to measure thyronine (To) in urine. The RIA used an anti-To antibody obtained from a rabbit immunized with a L-To-human serum albumin conjugate and [3H]To as the radioligand. The acetic acid analog of To (ToAc), that is the diphenyl structure with an acetic acid side-chain, cross-reacted strongly with the antibody. Relative to To, it cross-reacted 160% in phosphate-buffered saline, pH 7.4, and 100% in 0.075 mol/L barbital buffer, pH 8.6, containing sodium salicylate (final concentration, 8 mg/mL). The latter conditions were employed for the RIA, and the results reported thus reflect the presence of To and/or ToAc. 3-Monoiodothyronine, 3'-monoiodothyronine, 3',5'-diiodothyronine, and 3,5-diiodothyronine cross-reacted with the anti-To antibody 1.9%, 1.7%, 0.3%, and 0.2%, respectively; the cross-reactivity of other To derivatives and tyrosine and its derivatives was less than 0.05%. Urinary To and/or ToAc excretion in 12 normal subjects averaged 16 +/- 2 (+/- SE) micrograms/day (59 +/- 9 nmol/day) or 14 +/- 2 micrograms/g creatinine (5.9 +/- 0.6 nmol/mmol creatinine). Treatment of urine from normal subjects with beta-glucuronidase or sulfatase did not significantly alter the To content. Column and thin layer chromatographic studies revealed that 83% and 61%, respectively (range, 37-100%), of urinary To immunoreactivity was attributable to ToAc. The mean daily excretion of To in 20 patients with nonthyroidal illness [NTI; 22 +/- 4 micrograms/day (82 +/- 17 nmol/day)] was similar to that in normal subjects, but was elevated when expressed as nanomoles per mmol creatinine (20 +/- 2; P less than 0.001), because creatinine excretion was reduced in the NTI patients. The mean daily urinary To excretion in 13 patients with hyperthyroidism due to Graves' disease was slightly elevated [29 +/- 6 micrograms/day (108 +/- 21 nmol/day); P less than 0.1], but was clearly elevated when expressed as nanomoles per mmol creatinine (37 +/- 8; P less than 0.001), again because creatinine excretion was reduced in these patients. The mean urinary To excretion was subnormal in 13 patients with hypothyroidism and was significantly (P less than 0.005) less than that in the NTI patients regardless of the manner in which the results were expressed. Analysis of pronase hydrolysates of thyroid glands obtained at autopsy from euthyroid patients suggested that the To content of the thyroid approximates only 1.2% that of T4, supporting the thesis that prior iodination of tyrosine is critical for the coupling process in the thyroid.(ABSTRACT TRUNCATED AT 400 WORDS)

Epinephrine-induced alterations in urinary cyclic AMP in hyper- and hypothyroidism.

Although ratios of urinary cyclic AMP (cAMP) to creatinine were found in this study to be elevated in hyperthyroidism, as previously reported, this elevation appears to result primarily from a decrease in the rate of urinary creatinine excretion associated with the hyperthyroid state and not to be due to an increase in the urinary cAMP production rate. Indeed, there was no significant alteration observed in the urinary cAMP excretion found in 15 hyper-, 12 eu-, and 5 hypothyroid subjects. However, a slight, but significant increase in the 24-hour urinary cAMP excretion was noted in ambulating hyperthyroid subjects (8.5 +/- 2.4 muMol/day; normal 5.2 +/- 1.6 muMol/day; P less than .05). In contrast, the effect of the infusion of 0.05 mug/kg/min of epinephrine over a 2-hour period, resulted in a significantly greater rise in urinary cAMP excretion in hyperthyroid patients (0.83 +/- 0.07 muMol/h) compared to euthyroid subjects (0.53 +/- 0.4 muMol/h; P less than .005). Furthermore, hypothyroid subjects had no significant rise in urinary cAMP excretion after epinephrine infusion (P less than .001). Cardiovascular end-organ response to the epinephrine infusion was also greater in the hyperthyroid subjects and virtually absent in the hypothyroid group. These results suggest that there may be a significant alteration in the cAMP generating systems in states of thyroid hormone excess or insufficiency, and that provocative stimuli, such as epinephrine, may have its end-organ response modified by thyroid hormone effects on adenylate cyclase-cyclic AMP generating systems.

Urinary immunoprecipitation method for estimation of thyroxine to triiodothyronine conversion in altered thyroid states.

A new method is described for the estimation of T4 to T3 conversion in man and is applied to the study of hyperthyroid and hypothyroid clinical states. The method employs simultaneous iv injection of [125I]T4 and [131I]T3 with isolation of the labeled T3 tracers in 4- to 8-day pooled urine samples by a combination of solvent extraction, desalting, and immunoprecipitation procedures. Using [131I]T3 as a recovery standard, the T4 to T3 conversion ratio was found to be 0.470 +/- 0.011 in euthyroid subjects. This confirmed our earlier findings of 0.482 +/- 0.014 using a paper chromatographic method and nonsimultaneous isotope administration. The conversion ratio was increased in hypothyroidism to 0.535 +/- 0.011 (P less than 0.02) and decreased in hyperthyroidism to 0.415 +/- 0.009 (P less than 0.01). These changes parallel the fraction of the radioiodine collected in the urine for both T4 and T3; normal values are 77 +/- 4% for T4 and 76 +/- 4% for T3, values in hypothyroidism are 79 +/- 1% for T4 and 79 +/- 3% for T3, and values in hyperthyroidism are 58 +/- 3% for T4 and 58 +/- 5% for T3 (P less than 0.01). These findings indicate that 1) urinary T4 to T3 conversion values are highly reproducible in euthyroid as well as hyperthyroid and hypothyroid states; 2) the reduction in T4 to T3 conversion in hyperthyroidism probably reflects increased T4 disposal by nondeiodinative pathways and possibly the reverse in hypothyroid states; and 3) since urinary T4 to T3 conversion values in euthyroid subjects exceeded all reported conversion values in blood, there may be an alternate pathway of T3 production and disposal which is not reflected in the blood T3 production rate.