Platelet and adipocyte thermogenesis in hypothyroid patients: a microcalorimetric study

Direct microcalorimetry was used for measurements of heat production in cell suspensions of platelets and adipocytes, obtained from hypothyroid patients before and after 3 months on full L-thyroxine substitution. Platelet heat production was significantly lower than normal before treatment and increased in all 10 patients studied; the mean value increased from 51.3 +/- 1.6 fW/cell before to 57.1 +/- 1.8 fW/cell after therapy (P less than 0.001). Similarily, adipocyte heat production was initially significantly lower than normal and increased during treatment in all 6 patients investigated. The mean value for heat production per adipocyte was 18.8 +/- 1.7 pW/cell before and 32.4 +/- 2.5 pW/cell after therapy (P less than 0.025), which is still below the level recorded in lean healthy subjects. The adipocyte size did not change significantly. The increase in adipocyte heat production was correlated to the increase in S-triiodothyronine levels (r = 0.84, P less than 0.05). In hypothyroidism, the total metabolic activity seems to be comparatively more reduced in adipocytes than in platelets. A difference may exist between these cells with regard to recovery of normal metabolic activity during treatment for hypothyroidism. Direct microcalorimetry appears to be an adequate method for monitoring net metabolic effects of thyroid hormones in these cells.     

Erythrocyte thermogenesis in hyperthyroid patients: microcalorimetric investigation of sodium/potassium pump and cell metabolism

Erythrocyte thermogenesis was studied by microcalorimetry in 11 patients before and after treatment for hyperthyroidism. Cell heat production rate and intracellular Na+ and K+ levels were measured in plasma suspensions of erythrocytes with and without specific inhibition of Na/K ATPase by ouabain. The ouabain induced change in the heat production rate (the Na/K pump thermal power); the erythrocyte intracellular Na+ content and the ouabain sensitive Na+ transport were used to estimate the Na/K pump function. The mean value for heat production rate was 131 +/- 4 mW/L erythrocytes before treatment, which is significantly higher than in euthyroid subjects. A significant decrease (P less than 0.01) to normal levels was recorded following therapy. This decrease, as determined in samples with ouabain, correlated to changes in serum levels of triiodothyronine, T3, (r = .74, P less than 0.01). The Na/K pump thermal power was 11 +/- 2 mW/L erythrocytes (8 +/- 2% of total heat production rate) before and 9 +/- 2 mW/L erythrocytes (8 +/- 2%) after treatment. These two values were not different from those obtained in euthyroid subjects. The erythrocyte Na+ content decreased from 9.9 +/- 2.1 to 4.9 +/- 0.5 mmol/L erythrocytes (P less than 0.001) following normalization of thyroid function. The decrease in intracellular Na+ concentration correlated to the decrease in serum T3 levels, but only when calculated from the data obtained in samples with ouabain (r = .60, P less than 0.05). The relative increase in intracellular Na+ concentrations following addition of ouabain was significantly lower (P less than 0.05) before than after treatment for hyperthyroidism, 37 +/- 10% and 61 +/- 5%, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)     

Thyroid hormones and thermogenesis: The metabolic cost of food and exercise

To mimic plasma T₃ levels observed in a previous overfeeding study, six lean healthy men received replacement amounts of L-thyroxine (200 μg/d) to block endogenous thyroid hormone production while consuming their habitual diet. After 4 weeks equilibration on T₄, L-triiodothyronine (T₃) was given (45 μg/d) in addition to T₄, to produce mild T₃-thyrotoxicosis, for another 2 weeks. At the end of this period T₃ was discontinued but the subjects continued to receive T₄ for another 2 weeks. Resting metabolic rate, exercise efficiency, and the thermic effect of food were measured using a ventilated hood, open circuit indirect calorimeter at the end of each phase of the experiment. There was a significant increase in the resting metabolic rate of 6% (P<0.01) while the subjects were mildly T₃-thyrotoxic. The increase in energy expenditure however, during exercise on a bicycle ergometer or following a 500 kcal liquid-formula meal remained unaltered in the same situation. Thus, mild T₃-toxicosis does not alter the efficiency of exercise or the thermic effect of food. These results suggest that the increased plasma T₃ levels, observed in overfeeding, could explain corresponding increases in resting metabolic rate but not changes in the efficiency of exercise or the utilization of food.     

Serum free thyroid hormones in different degrees of hypothyroidism and in euthyroid autoimmune thyroiditis

Serum total and free T4 and T3, thyroxine-binding globulin (TBG) and TSH, basal and 20, 30 and 60 min after TRH (200 micrograms, iv), were evaluated in 125 hypothyroid patients (38 with severe, 23 with mild, and 64 with subclinical hypothyroidism), in 35 euthyroid subjects with autoimmune thyroiditis, and in 51 healthy controls. T4/TBG and T3/TBG ratios were also calculated. A significant decrease in all indices of thyroid function except for T3 occurred simultaneously with a significant increase in basal and TRH-stimulated TSH levels from healthy subjects to subclinical hypothyroids, from subclinical to mild and from mild to severe hypothyroids; euthyroid patients with autoimmune thyroiditis did not differ from healthy subjects. All severe hypothyroid patients had low T4 as well as free T4 (FT4), free T3 (FT3), T4/TBG and T3/TBG ratios, but among mild and subclinical hypothyroids direct determination of FT4 and FT3 proved to be a better index of thyroid function than determination of T4 and T3 even after correction for TBG levels. FT4 was the most commonly abnormal index (19 of 23 subjects with mild and 14 of 64 with subclinical disease). Regression analysis showed that FT4, T4/TBG ratio, T4, and FT3 had a significant inverse correlation with TSH in hypothyroid patients. Discriminant analysis showed that among the thyroid parameters, FT4 is the variable which discriminates best between control subjects and the 3 groups of hypothyroid patients. These data extend previous reports and in a large series of patients confirm the biological meaning and the clinical value of direct measurement of serum free thyroid hormones in hypothyroidism.(ABSTRACT TRUNCATED AT 250 WORDS)     

Cell metabolic activity in acromegaly: a microcalorimetric study of lymphocyte metabolism

A discrepancy between the clinical impression of disease activity and basal serum levels of growth hormone is often seen in patients with acromegaly. A slightly better relation has been found to serum levels of IGF-I, but a technique for evaluation of cell metabolic activity in this disease is still missing. For this purpose we used microcalorimetry to determine heat production rate in lymphocytes from 15 patients with acromegaly. The mean heat production rate was 2.90 +/- 0.15 pW/cell, significantly higher than in 13 healthy subjects, 2.31 +/- 0.12 pW/cell (p less than 0.01). Heat production rates did not correlate significantly with basal growth hormone levels, but increased, in a statistically significant manner (p less than 0.001), in parallel with the score index used to evaluate the clinical activity of the disease. Using the technique of microcalorimetry we could thus demonstrate an increased metabolic activity at a cellular level in patients with acromegaly, a finding that is in accordance with the view that an increased cell metabolic activity is a component of the disease process in acromegaly.     

Thyroid dysfunction and serum lipids: a community-based study

OBJECTIVE: It is uncertain whether subclinical hypothyroidism (SCH) is associated with hypercholesterolaemia, particularly in subjects with SCH and serum TSH < or = 10 mU/l. Design, PATIENTS AND MEASUREMENTS: Cross-sectional study of 2108 participants in a 1981 community health survey in Busselton, Western Australia. Serum total cholesterol and triglycerides were measured in all subjects and high density lipoprotein cholesterol (HDL-C) measured (and low density lipoprotein cholesterol (LDL-C) calculated) in a subgroup of 631 subjects at the time of the survey. In 2001, TSH and free T4 concentrations were measured on archived sera stored at -70 degrees C. Serum lipid concentrations in subjects with thyroid dysfunction and euthyroid subjects were compared using linear regression models. RESULTS: In the group as a whole, serum total cholesterol was higher in subjects with SCH (N = 119) than in euthyroid subjects (N = 1906) (mean +/- SD 6.3 +/- 1.3 mmol/l vs. 5.8 +/- 1.2 mmol/l, P < 0.001 unadjusted, P = 0.061 adjusted for age, age(2) and sex). Serum total cholesterol was similarly elevated in subjects with SCH and TSH < or = 10 mU/l (N = 89) (6.3 +/- 1.3 mmol/l, P < 0.001 unadjusted, P = 0.055 adjusted for age, age(2) and sex). In the subgroup analysis, LDL-C was higher in subjects with SCH (N = 30) than in euthyroid subjects (N = 580) (4.1 +/- 1.2 mmol/l vs. 3.5 +/- 1.0 mmol/l, P < 0.01 unadjusted, P = 0.024 adjusted for age, age(2) and sex). LDL-C was significantly increased in subjects with SCH and TSH < or = 10 mU/l (N = 23) (4.3 +/- 1.3 mmol/l, P < 0.001 unadjusted, P = 0.002 adjusted for age, age(2) and sex). CONCLUSION: SCH is associated with increased serum LDL-C concentrations, which is significant after adjustment for age, age(2) and sex.     

Phenotypic analyses and concanavalin-A-induced suppressor cell dysfunction of intrathyroidal lymphocytes from patients with Graves' disease

The expression of phenotypic markers and Concanavalin-A-induced suppressor activity was compared among mononuclear cells isolated from thyroid glands and peripheral blood of thionamide-treated patients with hyperthyroid Graves' disease and peripheral blood from normal subjects. Intrathyroidal lymphocytes were obtained by two different methods (TG-1 and TG-2 cells), gradient centrifugation of supernatants of minced thyroid tissue and overnight culture of thyroid debris after mechanical disaggregation and enzymatic digestion, respectively. The percentages of CD3+ cells (all mature T cells) among peripheral blood and TG-1 and TG-2 cells from Graves' patients were similar, but the percentages of B1+ cells (pan B cells) among the TG-1 and TG-2 cells were markedly increased compared to that in peripheral blood. The percentages of CD4+ cells among the TG-1 and TG-2 cells were significantly less than that in peripheral blood. The percentages of CD4+2H4+ cells among CD4+ cells in TG-1 and TG-2 cells also were significantly less than that in peripheral blood. The percentage of CD4+4B4+ cells among CD4+ cells in thyroid glands was markedly higher than that in peripheral blood. The percentages of CD8+ cells and CD8+CD11b- cells (cytotoxic T cells) in thyroid glands were significantly higher than those in peripheral blood from Graves' patients and peripheral blood from normal subjects. The CD8+CD11b+ cells were subdivided into two subpopulations on the basis of CD8 antigen density. The percentage of dull CD8+CD11b+ cells (natural killer cells) among TG-2 cells was lower than that in peripheral blood, but there was no significant difference in bright CD8+CD11b+ cells (suppressor-effector T cells) between thyroid glands and peripheral blood. The percent suppression induced by Concanavalin-A in both TG-1 and TG-2 cells was significantly decreased compared with that in peripheral blood. These results suggest that impairment of suppressor cell activity and an increased number of B cells exist in thyroid glands of patients with Graves' disease compared to those in peripheral blood. It, thus, appears likely that both B cell hyperactivity and suppressor T cell dysfunction may induce excess production of autoantibodies in the thyroid glands of such patients.     

Serum free thyroid hormones in T3-toxicosis: a study of 35 patients

Few data are available on serum free thyroid hormone concentration in patients with T3-toxicosis. In this study total and free T4 and T3 and TBG were evaluated in 35 subjects with T3-toxicosis, including 12 with untreated Graves' disease, 5 Graves' patients on methimazole treatment, 13 with autonomous adenoma, 3 with multinodular goiter, and 2 with subacute thyroiditis. T4, T3, free T4 (FT4), free T3 (FT3) as well as calculated T4/TBG and T3/TBG ratios were significantly higher in patients than in 37 healthy controls. Serum FT4 levels above the normal range were found in 19 subjects with T3-toxicosis (9 with untreated and one with methimazole-treated Graves' disease, 6 with autonomous adenoma, 1 with multinodular goiter and 2 with subacute thyroiditis). These data, together with the few previous reports, indicate that high FT4 levels are present in about half of the patients with so called T3-toxicosis, and that this occurs more frequently in diffuse than nodular goiter. It is suggested that the term T3-toxicosis be used only for those subjects with normal total and free T4.     

Effects of thyroid hormones on cardiac structure: a tissue characterization study in patients with thyroid disorders before and after treatment.

Experimental evidence suggests an involvement of thyroid hormones in myocardial nonmyocyte component growth. We evaluated the possible role of thyroid hormones in myocardial remodeling by ultrasonic tissue characterization (videodensitometry) in 8 hyperthyroid patients, in 10 hypothyroid patients, and in 2 patients with thyroid hormone resistance syndrome (RTH), before, 60, and 120 days after treatment (T0, T60, T120), and in 10 age-matched euthyroids. According to a previously described procedure, the derived collagen volume fraction (dCVF%, an echocardiographic index estimating the collagen content) was predicted from the pixel-level frequency distribution width (broadband, Bb) of the selected echocardiographic images. Thyrotropin (TSH), free thyroxine (FT4), and free triiodothyronine (FT3) were assessed by immunometric method. QT interval dispersion (QTd) on basal electrocardiogram was measured as a marker of dyshomogeneous ventricular repolarization. At T0, Bb and dCVF% were normal in hyperthyroid and euthyroid patients, and slightly increased in RTH patients, whereas significantly higher values were found in hypothyroids. At T60, a significant reduction in Bb was observed in hypothyroids, with nearly normal dCVF% values. This trend was confirmed at T120 with complete normalization of echoreflectivity. No echoreflectivity changes were observed in hyperthyroid and RTH patients during treatment. QTd was significantly increased in hypothyroids at T0, while no significant differences were found among groups at T60 and T120. Because the different videodeonsitometric myocardial properties observed in hypothyroid versus hyperthyroid patients correspond to an increase of dCVF%, this study suggests that thyroid hormones exert an inhibitory effect on myocardial collagen synthesis in humans.     

Intermediary metabolism in a lizard, Calotes versicolor: role of thyroid hormones

In Calotes versicolor, thyroidectomy did not alter the blood glucose level, lactate dehydrogenase (LDH liver and heart), acid phosphatase (Ac.Pase liver and kidney), and alkaline phosphatase (Alk.Pase liver and kidney) activities; significantly decreased the activities of glucose-6-phosphatase (G-6-Pase liver and kidney), glutamic oxaloacetic transaminase (GOT liver and heart), glutamic pyruvic transaminase (GPT liver), and urea concentration (liver and kidney); and increased liver cholesterol when compared to sham-operated controls. Administration of L-thyroxine (L-T4) or triiodo-L-thyronine (L-T3) to thyroidectomized lizards significantly stimulated the activities of G-6-Pase, Ac.Pase, GOT and GPT, concentration of glucose and urea, and decreased the cholesterol level. While the activities of all the enzymes studied and cholesterol level remain unchanged, glucose and urea levels decreased and increased, respectively, in thyroidectomized animals treated with actinomycin D. Chloramphenicol treatment did not affect any of the parameters studied. Simultaneous injections of actinomycin D or chloramphenicol with L-T4 prevented the hormone-stimulated activities of Ac.Pase, GOT, and GPT while the activities of LDH, G-6-Pase, Alk.Pase, glucose, urea, and cholesterol levels remain unchanged.     

Erythrocyte heat production in human obesity: microcalorimetric investigation of sodium-potassium pump and cell metabolism

Changes in overall cellular metabolism, induced by specific inhibition of the Na-K-pump, were determined in erythrocytes from 33 normal and 25 obese subjects. Cellular metabolism was determined by measurement of heat production rates in erythrocytes suspended in plasma with and without the cardioactive glycoside, ouabain. Specific inhibition with ouabain induced the same decrease of the heat production rate in the two groups (14 +/- 5 mW/L for normal subjects and 13 +/- 5 mW/L for obese subjects). In neither group was there a correlation between the ouabain-inhibitable rate of metabolism and body weight. The present study results do not give support to the suggestion that a defect in the Na-K-pump activity would exist in the erythrocyte of human obese subjects and could not therefore be of importance in the pathogenesis of the disease.     

Thyroid hormones and atrial natriuretic hormone secretion: study in hyper- and hypothyroid patients.

Plasma atrial natriuretic hormone (ANH) values were evaluated in 28 hyperthyroid patients and in 11 hypothyroid patients and compared with 20 healthy subjects. In hyperthyroid patients plasma ANH basal levels were significantly (p less than 0.01) higher (14.2 +/- 1.6 pmol/l) than in controls (7.8 +/- 0.4 pmol/l) and in hypothyroid patients (6.4 +/- 0.3 pmol/l). No significant differences were found between controls and hypothyroid patients. The propranolol-induced decrease in heart rate in hyperthyroid patients did not significantly affect the plasma ANH values. Conversely, after the methimazole-induced euthyroidism a return within the normal range of ANH values was observed. The thyroxine replacement in hypothyroid patients determined a small but significant (p less than 0.05) increase in plasma ANH values. Observed data suggest that in humans thyroid hormones may influence plasma ANH concentrations independently of their effect on heart rate.     

Influence of thyroid hormones on gluconeogenesis from glycerol in rat hepatocytes: a dose-response study

The role of L-3,3'-5 triiodothyronine (T3), in a pathophysiological range, on gluconeogenesis from low concentration of glycerol (2 mmol/L), was investigated in isolated liver cells from 24-hour fasted rats either thyroidectomized, normal, or treated by a T3 dose ranging from 1, 5, or 10 micrograms/d/100 g body weight (BW) during 3 days to 50 micrograms during 7 days. Gluconeogenesis from glycerol was decreased by 63% in hypothyroid rats and increased by 35% in severely hyperthyroid rats. However, in cells from mild hyperthyroid rats no increase of gluconeogenesis was observed. Nevertheless, in mild hyperthyroidism, alpha-glycerophosphate (G3P) was significantly decreased and gluconeogenesis from glycerol was not inhibited by the addition of ethanol (10 mmol/L), both of which have a drastic effect in cells from thyroidectomized rats. The decrease of gluconeogenesis observed in cells from thyroidectomized rats was reversed by the addition of pyruvate (10 mmol/L). Thus, when the cells were in a "reduced state" (addition of ethanol) the differences between the group were magnified, and when the cells were in an "oxidized state" (addition of pyruvate) the differences were suppressed. These findings suggest that alteration of the capacity of reducing equivalents transfer from the cytoplasmic compartment to the mitochondria is the main mechanism by which mild hyperthyroidism can stimulate gluconeogenesis.     

Thyroid hormone inhibition of intermediary metabolism in dog thyroid slices stimulated by different agonists.

The role of thyroid hormones in a short loop feedback in the thyroid is controversial. This process was studied in dog thyroid slices stimulated by TSH, carbachol and phorbol esters. Incubation of thyroid slices with T3 and T4 for 1 hour inhibited the subsequent stimulation of glucose oxidation induced by carbachol and phorbol esters but not by TSH. T3 also inhibited the stimulation of 32P incorporation into phospholipids stimulated by these two agonists. Glucose oxidation stimulated by TSH, carbachol and 12-0-tetradecanoyl-phorbol-13-acetate (TPA) was inhibited by rT3 and the inhibition was not reversed by methimazole, which did abolish the inhibition induced by iodide, MIT and DIT. TSH stimulation of cAMP was not blocked by T3 or T4 but was by rT3 and MIT- and DIT. The mechanism of such inhibition appears to be complex, possibly involving formation of iodide from rT3, MIT and DIT but also dependent on the intact iodothyronine. Moreover, our data suggest that T3 and T4 exert their inhibition on the thyroid through the phospholipids cascade and this mechanism is probably independent on the release of iodide from these iodocompounds.     

Subclinical thyroid dysfunction and blood pressure: a community-based study

OBJECTIVE: Overt hypothyroidism and hyperthyroidism are associated with hypertension, but it is uncertain whether the same is true of subclinical hypothyroidism and hyperthyroidism. DESIGN, SUBJECTS AND MEASUREMENTS: Cross-sectional study of 2033 participants (aged 17-89 years) in the Busselton Thyroid Study who did not have a history of thyroid disease. Systolic blood pressure (SBP), diastolic blood pressure (DBP) and the prevalence of hypertension (defined as SBP >or=140 mmHg, DBP >or=90 mmHg or on treatment for hypertension) in subjects with thyroid dysfunction and euthyroid subjects were compared using linear regression models. Subjects with treated hypertension (N = 299) were excluded from analyses of SBP and DBP but included in analyses of hypertension prevalence. RESULTS: Mean SBP, DBP and the prevalence of hypertension did not differ significantly between subjects with subclinical hypothyroidism (N = 105) and euthyroid subjects (N = 1859), nor did they differ between subjects with serum TSH concentrations in the upper reference range (2.0-4.0 mU/l; N = 418) and those with TSH concentrations in the lower reference range (0.4-2.0 mU/l; N = 1441). The prevalence of hypertension was higher in subjects with subclinical hyperthyroidism than euthyroid subjects (prevalence odds ratio 2.8, 95% confidence interval 1.3-6.0 adjusted for age, age(2) and sex), but this was based on a small number of subjects with subclinical hyperthyroidism (N = 35). CONCLUSIONS: Subclinical hypothyroidism is not associated with hypertension. The observed association between subclinical hyperthyroidism and hypertension requires confirmation in a larger sample.