Association between residual thyroid carcinoma and diffuse hepatic uptake of 131I following radioiodine ablation in postoperative total thyroidectomy patients.

In patients with differentiated thyroid cancer (DTC) total or near-total thyroidectomy, postoperative 131I ablation, and thyroid suppression therapy are reported to be associated with fewer recurrences than other treatments. Many patients with DTC after total thyroidectomy and radioablation therapy have diffuse hepatic uptake of radioiodine, and its clinical importance is debated. Some investigators report that diffuse liver uptake correlates with uptake in the thyroid bed or the presence of metastatic thyroid cancer somewhere in the body, whereas others note no such correlation. The purpose of this research was to determine the clinical importance of diffuse hepatic uptake of radioiodine after 131I ablative therapy in patients with DTC. We retrospectively reviewed 141 posttherapy scans done in 118 patients with DTC. Patients had had total thyroidectomy and were hypothyroid when serum thyroglobulin (Tg) levels were obtained, and they were treated with 30 to 200 mCi of 131I. Scans were performed 3 to 21 days after radioablation therapy. Information was collected regarding the patients' age and gender, the interval between the ablation therapy and scan, uptake of radioiodine, serum thyroglobulin level, thyroid-stimulating hormone (TSH) level, thyroglobulin antibodies, TNM classification, mortality, and recurrence. Diffuse liver uptake was classified from 0 to 4 depending on hepatic brightness. Radioiodine scans were done to determine whether there was uptake in the thyroid bed or elsewhere. Statistical analyses included analysis of variance and Kaplan-Meier survival analysis. Diffuse hepatic uptake was observed (grades 1-4) in 96.4% of the patients; thus 3.6% had no hepatic uptake. There was no significant association between liver uptake and the uptake in the thyroid bed, the dose of 131I administered for ablation therapy, thyroglobulin levels, age, stage of the disease, presence of local or distant metastases, recurrence, or survival. Diffuse hepatic uptake was therefore not associated with residual normal thyroid or metastases as suggested by some but not all previous investigators.

A comparison of recombinant human thyrotropin and thyroid hormone withdrawal for the detection of thyroid remnant or cancer.

Recombinant human TSH has been developed to facilitate monitoring for thyroid carcinoma recurrence or persistence without the attendant morbidity of hypothyroidism seen after thyroid hormone withdrawal. The objectives of this study were to compare the effect of administered recombinant human TSH with thyroid hormone withdrawal on the results of radioiodine whole body scanning (WBS) and serum thyroglobulin (Tg) levels. Two hundred and twenty-nine adult patients with differentiated thyroid cancer requiring radioiodine WBS were studied. Radioiodine WBS and serum Tg measurements were performed after administration of recombinant human TSH and again after thyroid hormone withdrawal in each patient. Radioiodine whole body scans were concordant between the recombinant TSH-stimulated and thyroid hormone withdrawal phases in 195 of 220 (89%) patients. Of the discordant scans, 8 (4%) had superior scans after recombinant human TSH administration, and 17 (8%) had superior scans after thyroid hormone withdrawal (P = 0.108). Based on a serum Tg level of 2 ng/mL or more, thyroid tissue or cancer was detected during thyroid hormone therapy in 22%, after recombinant human TSH stimulation in 52%, and after thyroid hormone withdrawal in 56% of patients with disease or tissue limited to the thyroid bed and in 80%, 100%, and 100% of patients, respectively, with metastatic disease. A combination of radioiodine WBS and serum Tg after recombinant human TSH stimulation detected thyroid tissue or cancer in 93% of patients with disease or tissue limited to the thyroid bed and 100% of patients with metastatic disease. In conclusion, recombinant human TSH administration is a safe and effective means of stimulating radioiodine uptake and serum Tg levels in patients undergoing evaluation for thyroid cancer persistence and recurrence.

Thyroid hormone export in rat FRTL-5 thyroid cells and mouse NIH-3T3 cells is carrier-mediated, verapamil-sensitive, and stereospecific.

Export of L-T3 out of the cell is one factor governing the cellular T3 content and response. We previously observed in liver-derived cells that T3 export was inhibited by verapamil, suggesting that it is due to either ATP-binding cassette/multidrug resistance (MDR1/mdr1b) or multidrug resistance-related (MRP1/mrp1) proteins. To test this hypothesis we measured T3 export in FRTL-5, NIH-3T3, and rat hepatoma (HTC) cells that varied in expression of these proteins. FRTL-5 and NIH-3T3 cells were found to contain a T3 efflux mechanism that is verapamil inhibitable, saturable, and stereospecific. By contrast, T3 efflux in HTC cells was slow and unaffected by verapamil. Neither FRTL-5 nor NIH-3T3 cells express mdrlb, but all three cell types express mrpl, as assessed by immunoblotting. Overexpression of MDR1 in NIH-3T3 cells did not enhance verapamil-inhibitable T3 efflux. Photoaffinity labeling of FRTL-5 and NIH-3T3 cells with [125I]L-T3 revealed a labeled 90- to 100-kDa protein that was not present in HTC cells. Verapamil and excess nonradioactive L-T3, but not D-T3, inhibited labeling of this protein. The lack of correlation between T3 efflux and MDR1 and mrpl expression and the finding of a photoaffinity-labeled putative transport protein smaller than MDR1 or mrp1 protein (approximately 170 kDa) suggest that a novel protein is involved in the transport of T3 out of cells.

Evidence for an ATP-dependent bile acid transport protein other than the canalicular liver ecto-ATPase in rats.

BACKGROUND & AIMS: Canalicular secretion is rate limiting in overall blood-to-bile transport of bile acids. Studies using transfected cells have implicated the canalicular ecto-adenosine triphosphatase (ecto-ATPase) in adenosine triphosphate (ATP)-dependent bile acid transport. However, the structural features of this ecto-ATPase are not those anticipated for an in-to-out ATP-dependent transporter. The aim of this study was to explore the possible existence of an ATP-dependent bile acid transport mechanism distinct from ecto-ATPase. METHODS: Bile acid transport activity and ecto-ATPase expression were analyzed in primary rat hepatocytes, rat hepatoma HTC cells, and specially adapted HTC (HTC-R) cells using plasma membrane vesicles and Northern blot, slot blot, ribonuclease protection assay, and Western blot analyses. RESULTS: Plasma membranes isolated from HTC-R cells exhibited ATP-dependent taurocholate transport, which was many-fold greater than that in HTC cells. Hepatocytes showed the highest transport rates. Protein and RNA analyses showed very low expression of ecto-ATPase in HTC and HTC-R cells compared with hepatocytes. There was no difference between the two cell types at both the RNA and protein level. CONCLUSIONS: These findings show the presence in HTC-R cells and, apparently in hepatocytes, of one or more proteins other than the ecto-ATPase that mediate ATP-dependent transport of bile acids.

Iodine metabolism and thyroid physiology: current concepts.

Iodine plays a central role in thyroid physiology, being both a major constituent of thyroid hormones (THS) and a regulator of thyroid gland function. This review concerns those aspects of thyroid physiology in which significant advances have been made in recent years. We have known for decades that the thyroid gland concentrates iodine (I-) against an electrochemical gradient by a carrier-mediated mechanism driven by ATP. A similar I- uptake mechanism is found in other organs, including salivary glands, stomach, choroid plexus, and mammary glands, but only in the thyroid does TSH regulate the process. This past year saw a major advance with the cloning of the thyroid I- transporter. This development opens the way to an elucidation of the regulation of I- transport in the normal gland and in thyroid neoplasms that lack this property ("cold" nodules). All of the subsequent steps in TH biosynthesis, from oxidation and organification of iodide to the secretion of T4 and T3 into the circulation, are stimulated by TSH and inhibited by excess iodine. Recently, some of the regulatory mechanisms have been clarified. The function of the major TH-binding proteins in plasma is to maintain an equilibrium between extracellular and cellular hormone pools. Transthyretin, the principal T4-binding protein in cerebrospinal fluid, may play a similar role in the central nervous system. Although it generally is agreed that cellular uptake of TH is a function of the unbound (free) form of the hormone, there is evidence that certain TH-binding plasma proteins (i.e., apolipoproteins) may serve specific transport functions. The intracellular concentration of T3, the active TH, is determined by the rates of cellular uptake of T4 and T3, the rates of metabolic transformation, including conversion of T4 to T3, and the rate of T3 efflux. The latter has been assumed to be a passive process. However, recent studies by our group in San Francisco have shown that T3 is transported out of cells by a specific, saturable, verapamil-inhibitable mechanism. This T3 efflux system is widespread among cells from many tissues, and, at least in liver, modulates intracellular and nuclear concentration of the hormone and thereby influences TH action.

Nuclear imaging in the management of thyroid carcinoma.

The avidity of differentiated thyroid carcinoma for iodine is the basis for the use of radioiodine (131I) both for the detection and the treatment of recurrent thyroid cancer in patients following initial surgical treatment (thyroidectomy). Because recurrence of this type of cancer may be delayed for many years, long-term follow-up is needed. Nowadays such surveillance involves clinical assessment, monitoring of serum thyroglubulin, and, when indicated, whole-body imaging with 131I. Sensitivity of 131I imaging depends on proper preparation of the patient and careful attention to imaging technique. Interpretation of images requires knowledge of physiologic sites of radioiodine concentration and causes of artifacts. Because of the potential suppressive effect of the radiation from a diagnostic administration of 131I on the uptake of a subsequent therapeutic administration (so-called stunning of thyroid tissue) many centers limit the amount given for scanning to 2-3 mCi (74-111 MBq). Several tumor-seeking radioisotopic agents other than radioiodine have shown promise for improving the detection of metastases, and some of these agents offer a useful adjunct to 131I in the management of selected patients, particularly in those with suspected metastatic disease and negative 131I scans.

Thyroid hormone export regulates cellular hormone content and response.

Actions of thyroid hormones (THs) are determined by intracellular free hormone concentration. Here we report that enhanced TH extrusion via a saturable, cold-sensitive mechanism lowers intracellular TH and causes TH resistance in hepatoma cells. Since these cells overexpress multidrug resistance P-glycoproteins and TH extrusion and resistance are blunted by verapamil, P-glycoproteins may mediate this resistance. Verapamil-inhibitable TH efflux was also found in primary hepatocytes, cardiocytes, and fibroblasts. These findings demonstrate that TH extrusion can modulate TH availability and action in mammalian cells.

Indices of thyroid function and weight loss in human immunodeficiency virus infection and the acquired immunodeficiency syndrome.

Abnormalities of thyroid hormone levels have been reported in the acquired immunodeficiency syndrome (AIDS), but there has been debate as to whether they are appropriate for the clinical status of the patients. Inappropriate maintenance of circulating 3,3',5-triiodothyronine (T3) levels could contribute to weight loss. Although many patients with AIDS have a history of wasting, recent data indicate that prolonged periods of stable weight occur in AIDS and that short-term weight loss is present in a subset of patients with anorexia, many of whom have active secondary infection (AIDS-SI). Therefore we analyzed thyroid hormone levels in a cohort of subjects that have been characterized in terms of recent weight loss and caloric intake. Asymptomatic patients with human immunodeficiency virus infection (HIV+) had short-term stable weights, normal caloric intake, and normal serum T3 levels. In AIDS, average short-term weight was stable, caloric intake was normal, and T3 levels were decreased by 19%. In AIDS-SI, both short-term weight loss and anorexia were significant, and this group showed a 45% decrease in T3 levels. The free T3 (FT3) index was decreased by 30% in AIDS and by 50% in AIDS-SI. Free thyroxine (FT4) levels were decreased while thyroxine-binding globulin (TBG) capacity was increased in HIV+ and AIDS; TBG sialylation was unchanged. Thyrotropin (TSH) levels were slightly increased in AIDS, although levels remained within the normal range. 3,3',5'-triiodothyronine (rT3) levels were decreased in HIV+, AIDS, and AIDS-SI. Thus asymptomatic patients with HIV infection whose weight is stable maintain normal T3 levels.(ABSTRACT TRUNCATED AT 250 WORDS)

Hepatic uptake of 3,5,3'-triiodothyronine: electrochemical driving forces.

We used the multiple indicator dilution technique to assess the electrochemical forces driving uptake of 3,5,3'-triiodo-L-thyronine (T3) across the basolateral plasma membrane in the single-pass perfused rat liver. With the use of 4 g/dl albumin solutions, the influx and efflux clearances were 0.020 +/- 0.005 and 0.0049 +/- 0.0017 (SE) ml.s-1.g liver-1, respectively, indicating that the total T3 concentration at equilibrium should be about four times greater in cytoplasm than in plasma. However, when the influx and efflux clearances were divided by the unbound (free) T3 concentration in the perfusate and cytosol, they were not different (3.76 +/- 0.26 vs. 4.30 +/- 0.38 ml.s-1.g liver-1), indicating that the uptake process does not generate a gradient of unbound T3 across the plasma membrane. To further test whether T3 uptake is driven by the electrical potential difference across the plasma membrane, liver cells were depolarized by isosmotic replacement of perfusate chloride with gluconate. There was no effect on uptake or efflux. To test whether uptake is coupled to influx of sodium, perfusate sodium was replaced with choline. Although there was a modest decline in both the influx and efflux clearances, there was no change in their ratio, as would be expected for sodium-coupled active transport. These results indicate that uptake of T3 across the basolateral hepatocyte membrane occurs by passive diffusion. We found no evidence to support concentrative, active transport by either electrogenic or sodium-coupled mechanisms.

Deiodination and deconjugation of the glucuronide conjugates of the thyroid hormones by rat liver and brain microsomes.

Radioiodinated thyroxine (T4) glucuronide (T4G) and triiodothyronine (T3) glucuronide (T3G), paired with T4 or T3, were incubated at 37 degrees C for 2 hours in the presence of dithiothreitol and microsomes that had been prepared from euthyroid rat liver or hypothyroid rat brain tissues, as sources of type I and type II iodothyronine 5'-deiodinases, respectively. Incubations with boiled microsomes served as controls. The incubated supernatant was analyzed by high-pressure liquid chromatography (HPLC) for content of T4, T4G, T3, T3G, and combined T2 and T2G. The deiodination of T4G resulted from incubation with both liver and brain microsomes, but was somewhat less active than the deiodination of simultaneously incubated T4. All batches of microsomes studied also caused deconjugation of both T4G and T3G. The data are compatible with the hypothesis that T4G can serve as an alternate pathway for conversion of T4 to T3 in these tissues.

Thyroxine (T4) transport and distribution in rats treated with EMD 21388, a synthetic flavonoid that displaces T4 from transthyretin.

To test whether plasma transthyretin (TTR) might play a specific direct role in the transfer of T4 from the plasma to tissues, in vivo kinetic studies were performed in control rats and in rats treated with EMD 21388, a synthetic flavonoid that displaces T4 from TTR. The plasma disappearance curves of simultaneously injected [125I]T4 and [131I]albumin were analyzed to determine the rate constant for the transfer of T4 from the extracellular compartment to the rapidly exchangeable intracellular compartment (KE) and the steady state distribution ratio of T4 between the rapidly exchangeable intracellular compartment and the extracellular compartment (Imax/Emin). When rats were injected ip with EMD 21388 (2 mumol/100 g BW), the free T4 fraction in serum increased approximately 8-fold. This was due to displacement of T4 from TTR, as assessed by electrophoresis of serum proteins in the presence of [125I]T4. Concomitantly, both KE and Imax/Emin increased 6-fold in the treated rats. These results fail to confirm a major specific role for TTR in the transfer of T4 from the plasma to tissues. Instead, they are consistent with both the free hormone transport hypothesis and the free hormone hypothesis in this setting.

Inability to detect an inhibitor of thyroxine-serum protein binding in sera from patients with nonthyroid illness.

Sera from 111 patients hospitalized on acute-care wards (including 32 in the intensive care unit) were examined for the possible presence of inhibitors of thyroxine (T4)-serum protein binding in an assay employing equilibrium dialysis. In 38 of these sera, the unbound (free) T4 fraction was 50% or more higher than the free T4 fraction in a pool of normal sera. From the free T4 fraction in each of the 111 serum samples and the free T4 fraction in the pool of normal sera, the predicted free T4 fractions in mixtures (1:1) of each of these sera with the normal pool were calculated (assuming the absence of binding inhibitors) from the appropriate mass action equations. It was reasoned that a free T4 fraction in any mixture that exceeded this predicted value would indicate the possible presence of a binding inhibitor. (The normal pool was selected for having a low serum triglyceride concentration, to minimize in vitro generation of free fatty acids.) However, for the 111 serum samples studied, the free T4 fraction in the mixture exceeded the upper 95% confidence limit of this predicted value in only one case, and then just barely. Thus, evidence for an inhibitor of T4-serum protein binding in sera from patients with nonthyroid illness could not be found. Twenty-eight of the serum samples were also examined in a similar assay that employed ultrafiltration of undiluted serum instead of equilibrium dialysis. Evidence for an inhibitor of T4-serum protein binding similarly could not be found. Because part of the reason for postulating the existence of such a binding inhibitor has been the performance of the triiodothyronine (T3) resin uptake test in patients with nonthyroid illness, an alternative explanation for this phenomenon was sought. When thyroid hormone-binding globulin (TBG) was desialylated by treatment with neuraminidase, its avidity for T4 was markedly decreased, but its avidity for T3 was unchanged. Thus, if desialylated TBG circulates in patients with nonthyroid illness as previously reported, it could explain not only the low serum T4 concentrations despite near normal immunoreactive TBG concentrations, but also the poor performance of the T3 resin uptake test (where T4 binding capacity is overestimated) in these patients.

The effects of nonthyroid disease and drugs on thyroid function tests.

Serious nonthyroid illness and caloric deprivation, which so often accompany systemic illness, have diverse and still incompletely understood effects on thyroid hormone economy. We have discussed the pathophysiologic basis for the most common pattern of alterations in routine thyroid function tests: a decreased serum T3 concentration; normal or, in critically ill patients, a low total serum T4 level; and a normal free T4 concentration. Another, less frequent pattern (high total and free T4 with a normal serum T3) can be encountered transiently in the acutely ill medical or psychiatric patient. With the recent advent of sensitive assays for TSH and better methods for serum free T4, it is now possible to define more quickly and accurately the thyroid-metabolic status of most of these sick patients; the vast majority are euthyroid. Certain drugs confound the picture. The most important of these include dopamine and high-dose glucocorticoids, both of which suppress TSH secretion from the pituitary and may actually cause a state of central hypothyroidism. Other drugs have multiple effects on thyroid hormone indices (e.g., amiodarone). Knowledge of all of the ways in which systemic illness, starvation, and certain drugs may influence thyroid function tests is crucial in assessing the thyroid status of patients with serious nonthyroid disease.

Iodide-induced hypothyroidism in patients after thyroid resection.

The purpose of this investigation was to determine whether an intrinsic defect in thyroid hormone production is required for the development of iodide-induced hypothyroidism or does it also develop in TSH-stimulated normal thyroid tissue. To answer this question, we studied the response to iodine administration (180 mg iodide daily for 3-4 months) in eight euthyroid patients who had had partial thyroidectomies 2 months to 10 years previously for benign thyroid nodules, and in three euthyroid control subjects. In all 11 euthyroid patients, basal serum TSH concentrations increased during iodide administration. In six of the eight patients who had previous thyroid operations and in two of the three control patients, basal serum TSH concentrations increased into the abnormal range (greater than 6 U ml-1). Increased serum TSH concentrations were noted as early as 1 week after potassium iodide had been started and the increased levels persisted during the period of iodide administration. Although basal values for serum TSH concentration were initially within the normal range, those patients with highest basal serum TSH values developed the greatest increase in TSH in response to potassium iodine. Among the eight patients treated by partial thyroidectomy, serum T4 concentrations decreased in five, serum T3 concentration decreased in three and all five developed mild symptoms of hypothyroidism while receiving iodide. Serum T4 concentrations also decreased slightly in two of the three control patients. Serum total iodine levels increased from 7.0 +/- 0.5 to 315.7 +/- 108.6 g dl-1 (mean-+/- standard error) during potassium iodide administration, but there was no correlation between the level of serum iodide concentration achieved and inhibition of thyroid function.(ABSTRACT TRUNCATED AT 250 WORDS)

Cyclic adenosine 3',5'-monophosphate and glucose stimulate thyroxine 5'-deiodinase type II in cultured mouse neuroblastoma cells.

Nutrient modulation increases mouse neuroblastoma (NB) T4-5'-deiodinase II (T4-5'-D II) activity. Carbohydrates are more potent than either amino acids or glycerol as nutrient sources. Glucose rapidly (2 to 4 hours) enhances NB enzyme activity and the response is dependent on new protein synthesis. The present study was performed to further characterize this glucose effect and explore its relationship to the cyclic adenosine monophosphate (cAMP) system in these cells. NB T4-5'-D II activity reached a maximum level (sixfold) in response to glucose (10 mmol/L) at 16 hours and thereafter remained constant up to 22 hours before reverting back to basal level between 24 and 30 hours. This pattern of response allowed the performance of detailed studies on maximum glucose activated NB T4-5'-D II under transient equilibrium conditions during the 16- to 22-hour period. Addition of dibutyryl cAMP (dbcAMP) (1 mmol/L) at this stage significantly increased enzyme activity (twofold at 2 hours and fourfold at 4 and 6 hours) compared with glucose alone. There was an additive response to dbcAMP under these maximum glucose-activated conditions. Nonactivated NB T4-5'-D II showed a twofold response to dbcAMP (1 mmol/L) at 4 hours in a glucose-free medium. Under these conditions, glucose (10 mmol/L) also increased enzyme activity twofold. Combined studies with dbcAMP and glucose increased enzyme activity fourfold at 4 hours. Subsequent studies were performed with forskolin (10 mumol/L) and cholera toxin (1 nmol/L), modulators of endogenous cAMP.(ABSTRACT TRUNCATED AT 250 WORDS)

Carbohydrate reactivation of thyroxine 5'-deiodinase (type II) in cultured mouse neuroblastoma cells is dependent upon new protein synthesis.

The T3 concentration in brain predominantly reflects local production from T4 rather than T3 uptake from the circulating pool. We recently demonstrated that rat brain T3 content is increased by glucose feeding compared to chow feeding. One possible mechanism for this effect is an increase in brain T4 5'-deiodinase (5'-D) activity. Our recent preliminary studies of neuroblastoma (NB) cells demonstrate that renewal of RPMI-1640 medium stimulates T4 5'-D type II (NB T4 5'-D II) activity in these cells. The present studies were performed to determine the mechanism of this response. Studies were performed on NB cells supported in thyroid hormone-depleted (deficient) medium. This approach increased NB T4 5'-DII activity 4-fold compared to that in thyroid hormone-replete medium. Medium renewal further stimulated enzyme activity (7- to 9-fold; maximum at 6 h) in each group. The difference between the hypothyroid group and control was sustained over a 24-h period. Subsequent studies demonstrated that glucose (11 mM) was the specific medium ingredient mediating the medium renewal response. A progressive increase in NB T4 5'-DII activity was noted over 8 h during RPMI-1640 salt plus glucose (11 mM) incubation. This was equivalent to the effect of complete medium containing glucose (11 mM). Coincubation with insulin (10(-7)-10(-9) M) did not modify the enzyme response to glucose. In addition, fructose (10 mM) had a similar effect on enzyme activity. Glycerol and essential and nonessential amino acids also modestly increased NB T4 5'-DII activity compared to that in the control group (P less than 0.01). Actinomycin-D (1 microM), cycloheximide (100 microM), and puromycin (100 microM) significantly (P less than 0.001) decreased the glucose effect on T4 5'-DII by 5-, 9-, and 17-fold, respectively, after 6 h of incubation. In addition, puromycin (10-200 microM) inhibited both NB T4 5'-DII activity and [3H]amino acid incorporation during incubation in glucose. There was a significant correlation between these parameters (r = 0.8; P less than 0.001). The enzyme activity decay curves in the glucose-activated and control groups subsequent to puromycin (100 microM) addition at 8 h were parallel. The fractional turnover rate was 13%/h in the controls and 11%/h in the glucose groups. The calculated enzyme production rate was significantly higher (P less than 0.005) in the glucose group compared to that in the control group (17.4 vs. 6.8 fmol/mg protein.h).(ABSTRACT TRUNCATED AT 400 WORDS)

Uptake of cortisol by the perfused rat liver: validity of the free hormone hypothesis applied to cortisol.

The mechanism by which cortisol in plasma enters hepatic cells was investigated using the isolated perfused rat liver. To determine whether hepatic uptake of cortisol from serum can be accounted for entirely by the pool of unbound (free) cortisol, we compared observed uptake rates with the equilibrium-free fraction of cortisol in serum and the rates of dissociation of cortisol from its serum binding proteins (determined using a rapid filtration assay based on transfer of [3H] cortisol to dextran-coated charcoal). More than 95% of the cortisol in both human and rat serum dissociated spontaneously from its binding proteins within 5 sec at 37 C. The fractional unidirectional hepatic uptakes of cortisol from pooled human serum and pooled rat serum were 59.4 +/- 5.4% and 59.5 +/- 1.0% (mean +/- SE), respectively, at the physiological flow rate of 1 ml/min.g liver. The corresponding free cortisol fractions in these sera were 4.53 +/- 0.15% and 8.16 +/- 0.23%, respectively. The fractional unidirectional hepatic uptake of cortisol from protein-free buffer averaged 99.9% (n = 5) at a flow rate of 3 ml/min.g liver. By calculating the appropriate rate constants and applying the Kety-Renkin-Crone equation to the above data, it can be shown that all of the cortisol taken up from serum by the perfused rat liver can be accounted for by the pool of free cortisol, which turns over very rapidly. The physiological significance of this finding is discussed in terms of a general mathematical model of hormone transport that delineates the conditions under which the free hormone hypothesis is and is not valid.

Thyroxine transport and distribution in Nagase analbuminemic rats.

The postulate that thyroxine (T4) in plasma enters tissues by protein-mediated transport or enhanced dissociation from plasma-binding proteins leads to the conclusion that almost all T4 uptake by tissues in the rat occurs via the pool of albumin-bound T4 (Pardridge, W. M., B. N. Premachandra, and G. Fierer. 1985. Am. J. Physiol. 248:G545-G550). To directly test this postulate, and to test more generally whether albumin might play a special role in T4 transport in the rat, we performed in vivo kinetics studies in six Nagase analbuminemic rats and in six control rats, all of whom had similar serum T4 concentrations and percent free T4 values. Evaluation of the plasma disappearance curves of simultaneously injected 125I-T4 and 131I-albumin indicated that the flux of T4 from the extracellular compartment into the rapidly exchangeable intracellular compartment was similar in the analbuminemic rats (51 +/- 21 ng/min, mean +/- SD) and in the control rats (54 +/- 15 ng/min), as was the size of the rapidly exchangeable intracellular pool of T4 (1.13 +/- 0.53 vs. 1.22 +/- 0.36 micrograms). This latter finding was confirmed by direct analysis of tissue samples (liver, kidney, and brain). We also performed in vitro kinetics studies using the isolated perfused rat liver. The single-pass fractional extraction by normal rat liver of T4 in pooled analbuminemic rat serum was indistinguishable from that of T4 in pooled control rat serum (10.9 +/- 3.3%, n = 3, vs. 11.4 +/- 3.4%). When greater than 98% of the albumin was removed from normal rat serum by chromatography with Affi-Gel blue, the single-pass fractional extraction of T4 (measured by a bolus injection method) did not change (16.3 +/- 2.1%, n = 5, vs. 15.2 +/- 2.5%). These data provide the first valid experimental test of the enhanced dissociation hypothesis and indicate that there is no special, substantive role for albumin in T4 transport in the rat.

The acute effects of human growth hormone administration on thyroid function in normal men.

GH replacement therapy may lead to alterations in serum TSH and/or thyroid hormone values in GH-deficient patients, but there is no consensus on the explanation for these changes. We examined the effect of GH administration (0.125 mg, sc, daily for 4 days) on thyroid function in 20 normal men. Serum T4 levels decreased by 8%, and serum free T4 index values decreased by 5%. In contrast, serum T3 levels increased by 21%; serum rT3 did not change. These changes were accompanied by a 54% decrease in the mean serum TSH level. While it is not possible to draw conclusions about hormone production and disposal rates from changes in serum levels, these data are most consistent with enhanced extrathyroidal (including intrapituitary) conversion of T4 to T3 and a compensatory decrease in TSH secretion.