Fraction Of T4 Research Articles

Analysis of the radiation effect on the activity of thyroid blood transport systems: connection with the features of thyroid status

Thyroid-binding proteins play an important role in regulating thyroid status. However, the question of the radiation effect on the functional state of thyroid blood transport systems remains insufficiently studied. The aim of the work was to assess the indicators of thyroid status and thyroid blood transport in persons exposed to radiation as a result of the Chernobyl accident. The task of the work was also to analyze the results of an oral test for thyroxine pharmacokinetics in patients with thyroid carcinoma who received radioiodine therapy. Screening studies conducted 5−6 years after the Chernobyl accident indicate the radiation effect on the free thyroxine content in the blood and the functional activity of thyroid protein transport in children and adolescents living in the territories of the Khoiniki district of the Gomel region. The dose-dependent effect of an absorbed dose of 131I was also detected in children and adolescents displaced from the 30 km zone of the Chernobyl accident. The results of an oral test for thyroxine pharmacokinetics demonstrate an increase in the free T4 content against the background of a stable total T4 fraction in patients with thyroid carcinoma who received high dose loads as a result of radioiodine therapy. The conducted studies confirm the possibility of the influence of a high dose of 131I on thyroid blood transport systems and on the ratio of free and bound fractions of thyroid hormones in the blood.

Antioxidant and a-amylase Inhibitory Activities and Phytocompounds of Clausena indica Fruits.

Background: Clausena indica fruit is commonly used for food ingredients and traditional medicines in tropical countries, however, information about its biological activities and chemical profiles has been inadequately reported. Methods: In this study, a bio-guided fractionation of antioxidants and α-amylase inhibitors from hexane (MH) and ethyl acetate (ME) extracts of C. indica fruit (pericarp and seed) was carried out. Eleven fractions from MH (D1–D11) and 17 fractions from ME (T1–T17) were obtained from column chromatography over silica gel, which were then examined for anti-radical capacity by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) assays, and pancreatic α-amylase inhibition, a key enzyme linked to type 2 diabetes. Results: Of isolated fractions, the fraction T4 revealed the most potent anti-DPPH activity (IC50 = 0.13 mg/mL), whereas T2 exhibits the strongest ABTS cation scavenging ability (IC50 = 0.31 mg/mL). In the enzymatic assay, the fractions D3 and T4 significantly inhibit the α-amylase reaction with IC50 values of 0.34 and 0.86 mg/mL, respectively. Remarkably, α-amylase suppression of T4 is close to acarbose and over four times stronger than palmitic acid, which are the well-known α-amylase inhibitors (IC50 = 0.07 and 1.52 mg/mL, respectively). The active constituents from fractions were identified by gas chromatography-mass spectrometry (GC-MS). The results show that the fraction D3 contains five major compounds, which are grouped in five classes consisting of fatty acids, phenols, benzodioxoles, alcohols, and sesquiterpenes. Among them, palmitic acid is the most dominant compound (32.64%), followed by 2R-acetoxymethyl-1,3,3-trimethyl-4t-(3-methyl-2-buten-1-yl)-1t-cyclohexanol (16.69%). Whilst, six major compounds belonging to fatty acid and coumarin classes are identified in the fraction T4. The most abundant compound in T4 is dentatin (47.32%), followed by palmitic acid (15.11%). Conclusions: This is the first finding that C. indica fruit can be a promising source for the development of natural antioxidant and antidiabetic agents. Additionally, the outcome reveals that dentatin, a known natural antineoplastic agent, can be feasibly exploited from C. indica fruit.

Characteristics and prognosis of interval cancers after biennial screen-film or full-field digital screening mammography.

We determined the characteristics and prognosis of interval breast cancers (IC) at screen-film (SFM) and full-field digital (FFDM) screening mammography. The study population consisted of 417,746 consecutive screening mammograms (302,699 SFM screens and 115,047 FFDM screens), obtained between 2000 and 2011. During 2-year follow-up, we collected breast imaging reports, surgical reports, and pathology results. A total of 800 ICs had been diagnosed in the screened population, of which 288 detected in the first year (early ICs) and 512 in the second year (late ICs) after a negative screen. 31.3% of early IC's and 19.1% of late IC's, respectively, were visible in retrospect on the latest previous screens, but had been missed during screening (P<0.001). Missed invasive ICs were larger (28.5mm vs. 23.9mm, P=0.003) and showed a higher fraction of T3+cancers (16.9 vs. 8.5%, P=0.02) than true ICs (i.e., not visible at the latest screen). A higher portion of missed than true ICs underwent mastectomy (44.7 vs. 30.8%, P=0.002). We found no differences in mammographic and tumor characteristics for early ICs, detected either after SFM or FFDM. Late ICs following FFDM were more often true ICs than missed ICs (69.0 vs. 57.6%, P=0.03) and more often receptor triple negative (P=0.02), compared to late ICs at SFM. Interval cancer subgroups showed comparable overall survival. Interval cancer subgroups show distinctive mammographic and tumor characteristics but a comparable overall survival.

Indicators of pituitary-thyroid system and lipid metabolism in female representatives of the Buryat ethnos and Europeoids

Characteristic features of indicators of the thyroid status and lipoprotein content were found in blood serum of girls and women representing Buryat ethnos and europeoids. In representatives of Buryat ethnos, as compared to europeoids, a development of adaptive reactions from the side of pituitary-thyroid part of the neuroendocrine regulation system was noted (increased concentrations of free T3 fraction in girls and decreased TTH levels in women). Changes in lipid metabolism parameters lie in lower concen- trations of atherogenic fractions of cholesterol in Buryat girls and increased level of cholesterol-contai- ning blood components in adolescent europeods. At older age period a reverse tendency is observed, cha- racterized by an increased activity of lipid metabolism in representatives of the indigenous nationality.

A role for dynamic contrast-enhanced magnetic resonance imaging in predicting tumour radiation response.

Background:Dynamic contrast-enhanced (DCE) MRI may provide prognostic insights into tumour radiation response. This study examined quantitative DCE MRI parameters in rat tumours, as potential biomarkers of tumour growth delay following single high-dose irradiation.Methods:Dunning R3327-AT1 prostate tumours were evaluated by DCE MRI following intravenous injection of Gd-DTPA. The next day tumours were irradiated (single dose of 30 Gy), while animals breathed air (n=4) or oxygen (n=4); two animals were non-irradiated controls. Growth was followed and tumour volume-quadrupling time (T4) was compared with pre-irradiation DCE assessments.Results:Irradiation caused significant tumour growth delay (T4 ranged from 28 to 48 days for air-breathing rats, and 40 to 75 days for oxygen-breathing rats) compared with the controls (T4=7 to 9 days). A strong correlation was observed between T4 and extravascular-extracellular volume fraction (ve) irrespective of the gas inhaled during irradiation. There was also a correlation between T4 and volume transfer constant (Ktrans) for the air-breathing group alone.Conclusions:The data provide rationale for expanded studies of other tumour sites, types and progressively patients, and are potentially significant, as many patients undergo contrast-enhanced MRI as part of treatment planning.

Hipotiroidismo Secundário à Irradiação Cervical: Avaliação Sistemática da Função Tiroideia no Seguimento

Sometimes Thyroid dysfunction is an underestimated consequence of radiation exposure. The underlying mechanism is not clearly understood, but it is likely to be multifactorial. As so, the specific risk factors associated with the development of secondary radiotherapy hypothyroidism remain undefined. The direct irradiation of the thyroid gland may result more frequently in hypothyroidism. This is an irreversible condition, requiring lifelong treatment and monitoring. To evaluate the incidence of hypothyroidism in patients with head and neck cancer who underwent neck irradiation on an intensive or adjuvant basis and determine whether it is justifiable to integrate the monitoring of parameters for evaluation of thyroid function (TSH, free fraction T3 e free fraction T4 ) in this patient group on the Institutional follow up protocol. This is an observational, retrospective descriptive study, which comprises a group of 376 patients with head and neck cancer undergoing radiotherapy, on adjuvant or intensive basis, between the years 2007 and 2012, at Portuguese Institute of Oncology of Coimbra, E.P.E. 145 patients met all the inclusion criteria. Standardized rating scales for the definition of hypothyroidism have been used - LENT-SOMA scales (Late Effects Normal Tissues Subjective Objective Management Analysis). Grade 1 or higher was considered as a complication hypothyroidism. The analysis of a cohort of 145 patients was carried out. The most frequent tumor site was the larynx (26.9%). Thirty-two patients received adjuvant radiotherapy and 113 were treated with intensive schemes. The most used radiotherapy technique was intensity modulated (IMRT), performed in 86.2% patients. The overall incidence rate of hypothyroidism at 12 months was 18.6%. The diagnosis for hypothyroidism was done between 9 and 12 months after treatment radiotherapy in 51.9% of the patients with this complication. An analysis of the published literature, hypothyroidism is the most common thyroid complication after irradiation and affects a large percentage of patients undergoing cervical radiotherapy with curative intent. It has been shown that the risk of hypothyroidism increases with time after exposure, but the highest incidence appears to be in the first two years after treatment. From our analysis we obtained at 12 months, a rate of incidence of hypothyroidism of 18.6% (16.55% grade 1 and grade 2, 2.1%). This study found an incidence of hypothyroidism corresponding to the lowest values reported in other published studies. The short duration of follow-up considered in this analysis, may justify these results. With these results and their correlation with the available literature, the authors consider justified and recommend the inclusion of the systematic evaluation of thyroid function in the follow-up protocol of these patients.

Epigenetic Modulation of Gene Expression during Keratinocyte Differentiation

BackgroundEpigenetic modulation of gene expression occurs by various methods, including DNA methylation and histone modification. DNA methylation of specific genes may affect the chromatin structure, preventing access by the transcriptional machinery. Although gene expression is dramatically changed during keratinocyte differentiation, there is no evidence of epigenetic modulation during the process of epidermal stratification.ObjectiveWe investigated whether epigenetic modulation is involved in keratinocyte differentiation-specific gene regulation.MethodsWe used trypsin to produce epidermal fragmentation (named T1-T4) and performed a morphological analysis using hematoxylin-eosin stain and cytokeratin expression based on reverse transcription polymerase chain reaction. We then constructed a DNA methylation microarray.ResultsEach epidermal fragment showed morphological features of the epithelial layer. T1 represented the basal layer, T2 was the spinous layer, T3 was the granular layer, and T4 was the cornified layer. The level of the K14 proliferation marker was increased in the T1 fraction, and the level of K10 differentiation marker was increased in the T2-T4 fractions. Using a methylation microarray with the T1 and T4 fractions, we obtained many hypermethylated and hypomethylated genes from differentiated keratinocytes.ConclusionThe importance of epigenetic modulation in target gene expression during keratinocyte differentiation is identified.

Goiter epidemiology in South Tirol

To ascertain the prevalence of goitre among schoolchildren in South Tyrol (uncontrolled voluntary iodine salt prophylaxis), the size of the thyroid gland was determined by palpation and ultrasound in 1,013 of 1,224 children, aged 6 to 14 years (547 boys, 466 girls), selected by statistically criteria. At the same time, to assess thyroid function, total and free fractions of T3 and T4, and TSH were measured, while iodine supply was determined by measuring urinary iodine. Stage Ia goitre was diagnosed by palpation in 157 children, stage Ib in 18 and stage II in two (goitre prevalence of 1.97% according to WHO criteria). Mean thyroid volume as determined by ultrasound was 4.26 (0.8-23.4) ml. As expected, it increased with age and body surface area: it was 2.55 +/- 0.49 ml in 6-year olds, 3.83 +/- 0.42 ml in 10-year olds and 6.42 +/- 0.69 ml in 14-year olds. Iodine excretion (143.8 [4.1-984.8] micrograms iodine/g creatinine) indicated adequate iodine supply to the tested subjects. South Tyrol is thus, according to WHO criteria, no longer an endemic goitre area. The voluntary use of iodinated cooking salt and changed food habits have probably effected this change in goitre epidemiology.

A Direct Free Thyroxine (T4) Immunoassay with the Characteristics of a Total T4 Immunoassay

Direct free thyroxine (T(4)) measurements have been linked to both T(4)-binding serum protein concentrations and protein-bound T(4) concentrations. Whether this is evidence of a relationship to total T(4) concentrations has not been reported. We compared an analog-based direct free T(4) immunoassay and a total T(4) immunoassay. Each assay was applied to the fractions of serum T(4) obtained by ultrafiltration and equilibrium dialysis. Both were applied to serum-based solutions in which free T(4), T(4)-binding proteins, protein-bound T(4), and total T(4) were systematically varied, held constant, or excluded. Neither the free T(4) assay nor the total T(4) assay detected dialyzable or ultrafilterable serum T(4). Both assays detected and reported the T(4) retained with serum proteins. Both free and total T(4) results were related to the same total T(4) concentrations in the presence and absence of T(4)-binding proteins. Both results were similarly related to total T(4) concentrations when free T(4) was held constant while total T(4) was varied. Both were similarly related to a total T(4) concentration that was held constant while free T(4) progressively replaced protein-bound T(4). These free T(4) results, like total T(4) results, were unresponsive to a 500-fold variation in dialyzable T(4) concentrations. New experiments extend the characterization of a longstanding and incompletely characterized analog-based free T(4) immunoassay. These free T(4) measurements relate to total T(4) concentrations in the same way that total T(4) measurements do.

Domain Requirement for the Membrane Trafficking and Targeting of Syntaxin 1A

Syntaxin plays a key role in intracellular membrane fusion in eukaryotic cells. The function of syntaxin relies on its proper trafficking to and targeting at the target membrane. The mechanisms underlying the trafficking and targeting of syntaxin to its physiological sites remain poorly understood. Here we have analyzed the trafficking of syntaxin 1A in INS-1 and CHO cells. We have identified the transmembrane domain together with several flanking positive-charged amino acids as the minimal domain required for the membrane delivery. Interestingly, we found that SNARE motif-exposed syntaxin 1A mutants were retained in endoplasmic reticulum (ER) and failed to transport to the cell surface in the absence of SNAP-25, suggesting that the exposure of the SNARE motif causes ER retention and complexation with SNAP-25 helps the ER escape. Finally, our data propose two key roles for the H(abc) domain: to protect nonspecific interaction by masking the SNARE motif and to participate in the clustering of syntaxin 1A to the fusion sites in the plasma membrane.

Effects of deracoxib and aspirin on serum concentrations of thyroxine, 3,5,3′-triiodothyronine, free thyroxine, and thyroid-stimulating hormone in healthy dogs

To evaluate the effects of deracoxib and aspirin on serum concentrations of thyroxine (T4), 3,5,3'-triiodothyronine (T3), free thyroxine (fT4), and thyroid-stimulating hormone (TSH) in healthy dogs. 24 dogs. Dogs were allocated to 1 of 3 groups of 8 dogs each. Dogs received the vehicle used for deracoxib tablets (PO, q 8 h; placebo), aspirin (23 to 25 mg/kg, PO, q 8 h), or deracoxib (1.25 to 1.8 mg/kg, PO, q 24 h) and placebo (PO, q 8 h) for 28 days. Measurement of serum concentrations of T4, T3, fT4, and TSH were performed 7 days before treatment (day -7), on days 14 and 28 of treatment, and 14 days after treatment was discontinued. Plasma total protein, albumin, and globulin concentrations were measured on days -7 and 28. Mean serum T4, fT4, and T3 concentrations decreased significantly from baseline on days 14 and 28 of treatment in dogs receiving aspirin, compared with those receiving placebo. Mean plasma total protein, albumin, and globulin concentrations on day 28 decreased significantly in dogs receiving aspirin, compared with those receiving placebo. Fourteen days after administration of aspirin was stopped, differences in hormone concentrations were no longer significant. Differences in serum TSH or the free fraction of T4 were not detected at any time. No significant difference in any of the analytes was detected at any time in dogs treated with deracoxib. Aspirin had substantial suppressive effects on thyroid hormone concentrations in dogs. Treatment with high dosages of aspirin, but not deracoxib, should be discontinued prior to evaluation of thyroid function.

Hypothyroidism.

Hypothyroidism.

Influence of Adsorption and Deproteination on Potential Free Thyroxine Reference Methods

There is a need for consensus concerning reference methods to be used for calibration of commercial free-thyroxine (FT(4)) assays. Three different potential reference techniques were investigated for adsorption of T(4) to membrane materials, including any in vitro solid surfaces to which T(4) might adsorb, and for efficient separation of the T(4)-binding proteins from the free hormone fraction. We compared ultrafiltration with different commercial ultrafiltration units, ultrafiltration by dialysis tubing, equilibrium dialysis, and ultracentrifugation. We measured the adsorption to membranes and materials with L-[(125)I]T(4). Separation efficiency was determined by measuring the T(4)-binding protein albumin in the ultrafiltrate and the dialysate as well as in the supernatant from the ultracentrifugation with a double-antibody sandwich ELISA technique. We found a constant relationship between the amount of T(4) adsorbed to the dialysis or ultrafiltration membranes/materials and the initial T(4) concentration in HEPES buffer (protein-free medium). T(4) was considerably less adsorbed from serum than from HEPES buffer (P <0.001). Serum T(4) was less adsorbed upon ultracentrifugation than during dialysis and ultrafiltration (P <0.001). It was difficult to completely separate FT(4) from the binding proteins by ultrafiltration and ultracentrifugation. Separation by ultrafiltration depended on the material used. No investigated separation technique provides technically and theoretically correct separation of the free fraction of T(4) from the protein-bound fraction. Equilibrium dialysis seems to be the least compromised.

Influence of imipramine on the hypothalamic/pituitary/thyroid axis of the rat

The effects of the tricyclic antidepressant drug imipramine at different levels of the hypothalamic/pituitary/thyroid axis were investigated in the rat. Intraperitoneal (IP) treatment for 14 days with imipramine at 10 mg/kg, but not 2 mg/kg, reduced serum total thyroxine (T4) and triiodothyronine (T3). A similar decrease in serum total T4 was observed in thyroidectomized T4-treated rats, suggesting that imipramine treatment enhances T4 clearance instead of reducing T4 secretion. There were no parallel decreases in serum free T4 and T3 concentrations, due to the simultaneous increase in the free fractions of both T4 and T3 following imipramine treatment. In vitro experiments using equilibrium dialysis indicated that neither imipramine nor its metabolite desipramine directly influenced the binding of T4 or T3 to their transport proteins following addition to normal serum, suggesting an indirect effect of imipramine or desipramine on free hormone concentrations in vivo. Concentrations of T4 and T3 in the brain, liver, and heart were unaffected by imipramine treatment, suggesting that the drug did not affect cellular uptake and metabolism of T4 and T3. Serum concentrations of thyrotropin (TSH) were unaffected by imipramine pretreatment at either dose level, compatible with the fact that serum free T4 and T3 concentrations were not reduced. Moreover, there was no difference in thyrotrope responsiveness to stimulation by TSH-releasing hormone (TRH) and to inhibition by T4 and T3 in rat anterior pituitary cells cultured ex vivo for 18 hours from control and imipramine-treated rats. Furthermore, in vitro exposure of cultured rat anterior pituitary cells to imipramine and desipramine indicated that both agents decreased TSH secretion only at concentrations greater than 10−6 mol/L. These concentrations of imipramine and desipramine in the culture medium would exceed the free concentrations of these drugs see in vivo therapeutically. In addition, no direct effects of 10−6 mol/L imipramine or desipramine on the TSH response to TRH or to T3 were observed in vitro in cultured pituitary cells. A potential indirect effect of imipramine or desipramine on TSH secretion via altered hypothalamic control of thyrotropes does not seem likely, due to the lack of effect of imipramine treatment on serum TSH concentrations in imipramine-treated rats. In conclusion, imipramine treatment reduces serum total T4 and T3 in the rat, with enhanced clearance being the most likely explanation for the effect on T4. There was no evidence for altered tissue T4 or T3 concentrations or for altered thyrotrope function. The enhanced T4 clearance may explain the reduction in total T4 reported for imipramine-treated depressed patients. However, the effects of imipramine treatment on the transport of thyroid hormones in plasma need to be examined in more detail in patients, since interspecies differences in the nature of the transport proteins preclude extrapolation of the present results from the rat.

Possible role of corticosterone in the down-regulation of the hypothalamo-hypophysial-thyroid axis in streptozotocin-induced diabetes mellitus in rats.

We investigated the effects of diabetes mellitus on the hypothalamo-hypophysial-thyroid axis in male (R x U) F1 and R-Amsterdam rats, which were found to respond to streptozotocin (STZ)-induced diabetes mellitus with no or marked increases, respectively, in plasma corticosterone. Males received STZ (65 mg/kg i.v.) or vehicle, and were killed 1, 2 or 3 weeks later. At all times studied, STZ-induced diabetes mellitus resulted in reduced plasma TSH, thyroxine (T4) and 3,5,3'-tri-iodothyronine (T3). Since the dialyzable T4 fraction increased after STZ, probably as a result of decreased T4-binding prealbumin, plasma free T4 was not altered during diabetes. In contrast, both free T3 and its dialyzable fraction decreased during diabetes, which was associated with an increase in T4-binding globulin. Hepatic activity of type I deiodinase decreased and T4 UDP-glucuronyltransferase increased after STZ treatment. Thus, the lowered plasma T3 during diabetes may be due to decreased hepatic T4 to T3 conversion. Median eminence content of TRH increased after STZ, suggesting that hypothalamic TRH release is reduced during diabetes and that this is not caused by impaired synthesis or axonal transport of TRH to the median eminence. Hypothalamic proTRH mRNA did not change in diabetic (R x U) F1 rats during the period of observation, but was lower in R-Amsterdam rats 3 weeks after STZ. Similarly, pituitary TSH and TSH beta mRNA had decreased in R-Amsterdam rats by 1 week after STZ treatment, but did not change in (R x U) F1 rats. The difference between the responses in diabetic R-Amsterdam and (R x U) F1 rats may be explained on the basis of plasma corticosterone levels which increased in R-Amsterdam rats only. Hypothalamic TRH content was not affected by diabetes mellitus, but the hypothalami of diabetic rats released less TRH in vitro than those of control rats. Moreover, insulin had a positive effect on TRH release in vitro. In conclusion, the reduced hypothalamic TRH release during diabetes is probably not caused by decreases in TRH synthesis or transport to the median eminence, but seems to be due to impaired TRH release from the median eminence which may be related to the lack of insulin. Inhibition of proTRH and TSH beta gene expression in diabetic R-Amsterdam rats is not a primary event but appears to be secondary to enhanced adrenal activity in these animals during diabetes.

Steady-state regulation of whole-body thyroid hormone pool sizes and interconversion rates in hypothyroid and moderately T3-stimulated rats.

Steady-state regulation of whole-body T3 and T4 distribution and metabolism were directly evaluated and compared in hypothyroid, euthyroid, and in euthyroid rats moderately T3-stimulated by continuous infusion of 0.15 microg/day L-T3 per 100 g BW, thereby supplementing euthyroid T3 sources by two thirds. Our goal was to develop deeper insights into the hierarchy, quantitative adequacy, and sensitivity of this regulatory system, in response to these hormone production challenges in constant steady state. We used a novel whole-body steady-state experiment design model and data analysis approach, which entails nonexclusive whole-body homogenate extracts and blood collected after 7-day infusions of tracer T3 (T3*) or T4*, quantitatively analyzed chromatographically for T3*, T4* and metabolite* concentrations. Hormone regulation implications across the 3 groups were assessed by comparing (per 100g BW) total body T4 to T3 and T3 from T4 conversion rates (CR(4-3) and CR(3-4)), total body pool sizes (Qtot) and distribution volumes (V(D)), total body production (PR), or plasma appearance rates (PAR), plasma clearance rates (PCR), and elimination rates (k). In the hypothyroid rats, absolute production of T3 from T4 was only a fourth of that in euthyroids: CR(3-4) = 1.55 vs. 6.77 ng/h, but the percent (efficiency) of whole-body T4 converted to T3 was more than double that in euthyroids: %CR(4-3) = 45.4% vs. 21.0%, reflecting an effective doubling of type I and/or type II 5'-deiodinase activity on a whole-body basis in response to severe curtailment of thyroidal production. Whole-body T3 pools and T3 production and clearance rates were all about 2 to 3 times lower in hypo- than in euthyroids: minimum Qtot = 36.8 vs. 100 ng, V(D3) = 148 vs. 236 ml, PAR3 = 3.44 vs. 9.09 ng/h, PCR3 = 13.8 vs. 21.3 ml/h; and nearly all T4 pool size, production, clearance and elimination rates also were very substantially reduced: PCR4 = 0.540 vs. 0.941 ml/h, PR4 = 4.11 vs. 38.3 ng/h, Qtot4 = 128 vs. 702 ng, k4 = 0.0322 vs. 0.0530 h(-1). In moderately T3-stimulated rats, presumed central feedback effects of the added T3 on T4 production and total body pool size also were quite pronounced: PR4 = 21.4 ng/h and Qtot4 = 346 ng were reduced to about half that in euthyroids, but T4 elimination indices were virtually unchanged, and T3 production and elimination were minimally affected. Thus, overall, stabilizing negative feedback regulation of TH functioning at different hierarchical levels is quite bidirectionally sensitive. We found very tight (inhibitory) control over thyroidal T4 secretion, possibly also T3 secretion, and probably also absolute T3 production from T4, in response to moderate (+68%) supplements in T3 production; and the efficiency of total body T3 production from available T4 was amplified substantially in the severe primary hypothyroid state, although not nearly enough to compensate for the malady. Finally, the blood to total body pool fractions (Qb/Qtot) of both T3 and T4, but not the plasma or blood hormone levels, remained remarkably constant in response to these oppositely directed hormone production challenges, suggesting this ratio as an actively regulated, homeostatically-maintained entity.

Effects of long-term food reduction on the hypothalamus-pituitary-thyroid axis in male and female rats.

The reduced thyroid activity during short-term starvation is associated with a lowered hypothalamic synthesis and secretion of TRH. However, little is known about the cause of the reduced thyroid function during prolonged malnutrition. We have therefore studied the effects of food reduction to one-third of normal (FR33) on the hypothalamus-pituitary-thyroid axis of male and female Wistar rats. After 3 weeks body weights of FR33 rats were almost 50% lower than those of controls. In both sexes, FR33 caused marked increases in serum corticosterone, and decreases in serum TSH, thyroxine (T4), free T4, tri-iodothyronine (T3) and free T3. While the free T3 fraction (FFT3) in serum decreased, the free T4 fraction (FFT4) tended to increase. Electrophoretic analysis indicated that decreased FFT3 was correlated with an increased thyroxine-binding globulin, while the increase in FFT4 seemed due to a decreased thyroxine-binding prealbumin binding capacity. Total RNA and proTRH mRNA in the hypothalamus were not affected by FR33. Median eminence and posterior pituitary TRH content tended to increase in FR33 rats, suggesting that hypothalamic TRH release is reduced in FR33 rats. Anterior pituitary TSH content was decreased by FR33 in both sexes, but pituitary TSH beta mRNA and TRH receptor status were not affected except for increased pituitary TSH beta mRNA in female FR33 rats. Although FR33 had no effect on pituitary weight, pituitary RNA and membrane protein content in FR33 rats were 50-70% lower than values in controls. In conclusion, prolonged food reduction suppresses the pituitary-thyroid axis in rats. In contrast to short-term food deprivation, the mechanism whereby serum TSH is suppressed does not appear to involve decreases in proTRH gene expression, but may include effects on pituitary mRNA translation. Our results further support the hypothesis that TSH release may be lowered by increased corticosterone secretion, although the mechanism of this effect may differ between acute starvation and prolonged food reduction.

Effect of illness not associated with the thyroid gland on serum total and free thyroxine concentrations in cats

Objective To determine circulating concentrations and fate of total and free thyroxine (T4) in cats with various illnesses not associated with the thyroid glands (non-thyroidal illnesses). Design Prospective study. Animals 98 cats with nonthyroidal illness and 50 clinically normal cats. Procedure Serum total T4 concentrations were measured by radioimmunoassay, and serum free T4 concentrations were measured by direct equilibrium dialysis. Free T4 fraction was calculated from these 2 values. Results Serum total T4 concentrations were significantly (P &lt; 0.001) lower in sick cats (mean ± SD, 17.18 ± 8.14 nmol/L), compared with healthy cats (mean ± SD, 26.00 ± 7.62 nmol/L). Serum total T4 concentrations were inversely correlated with mortality. Differences in serum free T4 concentrations in sick cats (mean ± SD, 27.70 ± 13.53 pmol/L), compared with healthy cats (mean ± SD, 24.79 ± 8.33 pmol/L), were not significant. A few sick cats had serum free T4 concentrations greater than the reference range. Calculated free T4 fraction was significantly (P &lt; 0.001) greater in sick cats (mean ± SD, 0.24 ± 0.30%), compared with healthy cats (mean ± SD, 0.10 ± 0.06%). Clinical Implications Euthyroidism is maintained in sick cats, despite low serum total T4 concentrations. Measurement of serum total T4 concentrations is a valuable prognostic indicator. Serum free T4 concentrations should be used cautiously as a sole diagnostic criterion for confirmation of hyperthyroidism. (J Am Vet Med Assoc 1996;208:2004-2008)

Normal Serum Free Thyroid Hormone Concentrations in Patients Treated With Phenytoin or Carbamazepine

To address the paradox that phenytoin- and carbamazepine-treated patients have decreased serum free thyroxine (T4) and triiodothyronine (T3) concentrations but appear clinically euthyroid and have normal serum thyroid-stimulating hormone (TSH) concentrations. In vitro studies comparing measurements of total and free T4 and T3 by ultrafiltration assay (undiluted serum) and a commercial free T4 estimate kit in control serum samples or serum samples containing added therapeutic levels of phenytoin or carbamazepine. These measurements were made in serum samples diluted 1:5 with either identical serum or phospate buffer, pH 7.4, and in serum samples from patients with seizure disorders who were treated with phenytoin or carbamazepine. A 650-bed teaching hospital. Selected patients (n=19) who were in good health except for seizure disorder, with stable anticonvulsant drug levels in the upper half of the therapeutic range, and were not taking any other drugs that could affect thyroid parameters. Serum concentrations of free T4 and free T3 in patients taking phenytoin or carbamazepine vs normal controls. Addition of phenytoin or carbamazepine to normal human serum in vitro resulted in a significant increase in free T4 fraction and free T4 (P<.001). In patients taking phenytoin or carbamazepine, serum total T4 decreased significantly to 60% and 74%, respectively, of the control serum concentration (P<.001 for both phenytoin and carbamazepine); free T4 fraction (by ultrafiltration assay) increased 65% and 44%, respectively (P<.001 for phenytoin, P<.01 for carbamazepine); and free T4 remained unchanged. Free T4 concentration measured by a commercial kit (1:5 serum dilution) was significantly lower than the control concentration in both phenytoin- and carbamazepine-treated patients. Serum free T3 and serum TSH were also normal in phenytoin- and carbamazepine-treated patients. Therapeutic levels of phenytoin and carbamazepine displace T4 and T3 from serum binding proteins. When added to serum, the drugs effect an increase in free hormone fractions and free T4 and T3. In drug-treated patients, increased free T4 and T3 fractions offset the significant decrease in serum T4 and T3, resulting in normal free T4 and free T3 concentrations. Since currently available clinical tests will continue to show decreased free T4 concentrations in patients taking phenytoin or carbamazepine, clinicians should rely on serum TSH measurements to confirm the euthyroid status of these patients.

Uptake of triiodothyroacetic acid and its effect on thyrotropin secretion in cultured anterior pituitary cells

The uptake of [125I]triiodothyroacetic acid ([125I]Triac) in anterior pituitary cells was investigated and compared with that of [125I]T3. Furthermore, the effects of Triac, T3, and T4 on TSH release were compared. Cells isolated from adult male Wistar rats were cultured for 3 days in medium with 10% fetal calf serum. Uptake was measured at 37 C with [125I]Triac (100,000 cpm; 120 pM) or [125I]T3 (50,000 cpm; 50 pM) in medium with 0.5% BSA. In this medium, the ratio of the free fractions of Triac, T3, and T4 was 1:8:1. Exposure of cells to 100 nM TRH for 2 h stimulated TSH release by 80-110% (P < 0.001). Comparing total hormone levels (1 nM to 1 microM), Triac and T3 were equally effective in reducing this response, and both were 10-fold more effective than T4. The time course (15 min to 4 h) of [125I]Triac uptake was similar to that of [125I]T3, showing equilibrium after 1 h. Unlabeled Triac (1 microM) reduced the uptake of [125I]Triac and [125I]T3 at all time intervals. Expressed per pM free hormone, the cellular and nuclear uptake of [125I]Triac were twice those of [125I]T3. The 15-min uptake of [125I]Triac was reduced by incubation with 10 nM unlabeled Triac (35%; P < 0.001). Maximum inhibition (56%; P < 0.001) was found with 10 microM Triac. A similar effect was seen with 10 microM T3, T4, or 3,3',5,5'-tetraiodothyroacetic acid. Preincubation (30 min) and incubation (15 min) with 10 microM oligomycin reduced the cellular ATP content by 51% (P < 0.001), [125I]T3 uptake by 77% (P < 0.001), and [125I]Triac uptake by only 25% (P < 0.001). The temperature dependence of [125I]Triac and [125I]T3 uptake was the same. Preincubation and incubation with 10 microM monensin (reduces the Na+ gradient) or 10 microM monodansylcadaverine (inhibits receptor-mediated endocytosis) reduced 15-min [125I] Triac uptake by 15% (P < 0.005) and 19% (P < 0.005), respectively. The data show that 1) Triac, on the basis of the free hormone concentration, is more potent than T3 or T4 in suppressing TSH secretion; and 2) the rapid uptake of [125I]Triac by the anterior pituitary occurs by a carrier-mediated mechanism that is only partially dependent on ATP or the Na+ gradient.