T4 Release Research Articles

7951 The Response of HPT Axis & BAT to Cold Exposure Is Delayed in Female Rats Compared to Males

Abstract Disclosure: A. Gutiérrez-Mata: None. L. Jaimes-Hoy: None. I. Sotelo-Rivera: None. F. Romero-Arteaga: None. R.M. Uribe: None. J.L. Charli: None. P. Joseph-Bravo: None. Cold exposure activates the sympathetic nervous system releasing NE in thermogenic brown adipose tissue (BAT) and in parallel, the Hypothalamus-Pituitary-Thyroid (HPT) axis, increasing synthesis and release of hypothalamic-TRH, pituitary-TSH, and thyroid hormones T4 and T3; T4 in concert with NE induce brown adipose tissue (BAT) thermogenesis through stimulation of deiodinase 2 (Dio2), and uncoupling protein 1 (Ucp1) expression. These changes are detected within the first hour in male rats [J Neuroendo 12:861, 2014] while in females, only in ovariectomized [J Neuroendocrinol 21:439, 2009]. Given the thermogenic effect of ovarian hormones, we hypothesized that females could respond to longer exposures. Female Wistar rats were exposed individually to 5°C for 1 to 5h and compared to male’s acute response (15 min to 4h); controls were moved individually to a nearby room at 22°C for similar times. Rats were euthanized by guillotine, rectal temperature was immediately measured, trunk blood collected, serum separated, and tissues kept at -70°C. Hormone concentrations were quantified in serum by RIA and ELISA, and gene expression in paraventricular nucleus (PVN), anterior pituitary (AP), and BAT by RT-PCR or qPCR. Body temperature of cold-exposed males was higher than isolated controls since the first 30min, Trh expression increased at 1h in the paraventricular nucleus (PVN), TSH release from 15 min up to 2 hours, T4 levels peaked until 4h, and T3 increased after 2h as in [Endocrinology 58:140, 1993]; Trhr1 in AP reached its lowest levels at 2h. In BAT, Dio2 expression peaked at 1h and subsequently decreased. Ucp1 increased from 30 min and remained elevated until 4h, Adrb3, and Ppargc1a expressions were highest at 4h. Cold exposure in females decreased body temperature after 4 hours when Trh PVN expression increased and that of TRH receptor 1 (Trhr1) in AP was lowest; TRH degrading enzyme (Trhde) expression peaked after 2h of cold and subsequently decreased at 4h. TSH in serum increased at 2h and then returned to control levels, T4 slightly increased at 3h, and T3 peaked at 5h. Progesterone serum concentration increased from 2h and peaked at 4h. No changes were found in estradiol serum concentration. In BAT, Dio2 expression increased from 2 to 4h, Ucp1 at 3 and 5h while that of β3 adrenergic receptor (Adrb3) decreased at 2h but rose reaching a peak at 4h. In contrast, expression of Pgc1α -the major transcriptional coactivator of Ucp1- rapidly increased (1h) decreasing afterward. The data suggest that when the ambient temperature decreases, the sympathetic system is activated including stimulation of progesterone, and supports a delayed response of female HPT axis to cold exposure, compared to male’s leading to the proposal that when female body temperature diminishes, the HPT axis and thyroid hormone-induced thermogenesis are activated.Funding: DGAPA IN217422, PAEP, CONAHCYT 790605. Presentation: 6/2/2024

Dual-targeted nano-encapsulation of neonatal porcine islet-like cell clusters with triiodothyronine-loaded bifunctional polymersomes

There is growing evidence that neonatal porcine islet-like cell clusters (NPCCs) isolated from piglets can be used to treat type 1 diabetes in humans. However, graft rejection is a common complication in humans owing to the prevalence of xenoantigens in porcine. Therefore, researchers have investigated various islet encapsulation techniques that could protect against these antigens. To this end, this study presents a robust nano-encapsulation method based on bifunctional polymersomes (PSomes), in which N-hydroxysuccinimide (NHS) and maleimide (Mal) groups conjugated to the PSomes terminal interact with the amine and thiol groups on the surface of NPCCs to induce dual targeting via two covalent bonds. The findings indicate that the ratio of NHS to Mal on PSomes is optimal for dual targeting. Moreover, triiodothyronine (T3) is known to promotes pancreatic islet maturation and differentiation of endocrine cells into beta cells. T3 encapsulated in PSomes is shown to increase the glucose sensitivity of NPCCs and enhance insulin secretion from NPCCs. Furthermore, improvements in the nano-encapsulation efficiency and insulin-secreting capability of NPCCs through dual targeting via dual-Psomes are demonstrated. In conclusion, the proposed nano-encapsulation technique could pave the way for significant advances in islet nano-encapsulation and the imprevement of NPCC immaturity via T3 release.

SAT488 Wolff-Chaikoff’s Revenge: There-and-Back Again Graves Thyrotoxicosis

Abstract Disclosure: A. Akofu: None. S.A. Brietzke: None. N. Sheung: None. A. Uddin: None. N. Akabogu: None. S. Dejhansathit: None. F. marium: None. Grave’s disease is due to autoantibodies activating TSH receptors, driving downstream effects of iodine organification, iodotyrosine coupling, and release of T4 and T3 from intracellular stores. Severe thyrotoxicosis is traditionally treated with high dose thionamides to block thyroid hormone synthesis, iodides to slow iodine transport and block thyroid hormone release (collectively known as the Wolff-Chaikoff effect), and beta-blockers to slow heart rate and relieve tremulousness. We report the case of a health-conscious patient who had taken OTC iodide for years, and whose thyrotoxic condition abruptly worsened with discontinuation of his iodide supplement. A 67-year-old man was found in a delirious state wandering in the woods and was taken to the hospital ED. He had afib with RVR and evidence of high-output CHF. Diffuse goiter twice normal size was obvious on exam, and he was tremulous and diaphoretic. Serum TSH was <0.005 mcU/ml, with FT4 > 7.77 ng/dl and FT3 7.8 pg/ml. Initial treatment was metoprolol 25 mg TID, propylthiouracil (PTU) 300 mg TID, cholestyramine 4g BID and Lugol’s iodine 10 drops TID. Encephalopathy resolved after 2 days and FT4 decreased to 3.72 ng/dl after 8 days. Cholestyramine and Lugol’s iodine were thus discontinued. 2 days later, FT4 further declined to 2.75 ng/dl and he was switched from PTU to methimazole 30 mg twice daily. After 3 days, FT4 had increased to 5.17 ng/dl, and in response methimazole was increased to 40 mg TID. 3 days later FT4 increased further to 6.43 ng/dl, prompting a switch from methimazole back to PTU 300 mg TID. Because of further increase in FT4 and FT3, cholestyramine and Lugol's iodine were resumed. 3 days later, FT4 had declined from > 7.7 to 5.4 ng/dl, and he was discharged with plan to perform thyroidectomy once FT4 and FT3 reached nearly normal levels. Near the end of his hospitalization, he divulged that he had taken iodide drops for “thyroid health” for many years independent of medical advice, but hadn’t taken it for several days during the period of being lost in the woods prior to admission. Possible explanations for resistance of thyrotoxicosis to aggressive prescribed treatment include medication non-adherence, drug malabsorption, hyperaccelerated drug metabolism, anti-drug antibodies and antagonism of thionamide action by excessive intrathyroidal iodine. In this case, we hypothesize that his use of iodide serendipitously attenuated severity of Graves disease, by giving him a prolonged Wolf-Chaikoff effect. Discontinuation of iodide over a matter of days allowed reversal of this phenomenon and prevented control of his hyperthyroid condition until the deliberate restoration of the Wolf-Chaikoff effect with therapeutically prescribed Lugol’s solution. Whenever response to usual hyperthyroidism treatment is ineffective or paradoxical, as in this case patients should be specifically queried regarding use of OTC iodine preparations. Presentation Date: Saturday, June 17, 2023

Function of Pituitary-Thyroid Axis in Copper-Deficient Rats

Thirty male weanling Sprague-Dawley rats were fed either a copper-deficient diet (0.85 mg Cu/kg) or a copper-adequate diet (8 mg Cu/kg). After 7 weeks, the rats were fasted for 12 hours, and injected intravenously with thyrotropin-releasing hormone (TRH, 30 ng/100 g body weight). Six rats from each treatment were killed at 0, 0.5, 1, 2 and 4 hours after TRH injection. Sera obtained at 0, 0.5 and 1 hour, and at 0, 2 and 4 hours were used for the radioimmunoassay of thyroid-stimulating hormone (TSH) and thyroxine (T4), respectively. Reduction in liver copper content confirmed that the rats fed the test diet were copper-deficient. Serum TSH levels appeared to have peaked at 30 minutes but declined to a level higher than basal at 1 hour. No difference in TSH response was observed between the 2 treatments. Serum T4 response to TRH was reduced in the copper-deficient as compared to the adequate rats at all time intervals. After 2 hours a slight elevation was observed in the controls, but marked elevations in T4 were observed in both treatments at 4 hours. This reduction in T4 levels could be due to an impaired T4 synthesis or release in copper-deficient rats.copper deficiency thyroid-stimulating hormone thyroxine

A redox-reactive delivery system via neural stem cell nanoencapsulation enhances white matter regeneration in intracerebral hemorrhage mice.

Intracerebral hemorrhage (ICH) poses a great threat to human health because of its high mortality and morbidity. Neural stem cell (NSC) transplantation is promising for treating white matter injury following ICH to promote functional recovery. However, reactive oxygen species (ROS)-induced NSC apoptosis and uncontrolled differentiation hindered the effectiveness of the therapy. Herein, we developed a single-cell nanogel system by layer-by-layer (LbL) hydrogen bonding of gelatin and tannic acid (TA), which was modified with a boronic ester-based compound linking triiodothyronine (T3). In vitro, NSCs in nanogel were protected from ROS-induced apoptosis, with apoptotic signaling pathways downregulated. This process of ROS elimination by material shell synergistically triggered T3 release to induce NSC differentiation into oligodendrocytes. Furthermore, in animal studies, ICH mice receiving nanogels performed better in behavioral evaluation, neurological scaling, and open field tests. These animals exhibited enhanced differentiation of NSCs into oligodendrocytes and promoted white matter tract regeneration on Day 21 through activation of the αvβ3/PI3K/THRA pathway. Consequently, transplantation of LbL(T3) nanogels largely resolved two obstacles in NSC therapy synergistically: low survival and uncontrolled differentiation, enhancing white matter regeneration and behavioral performance of ICH mice. As expected, nanoencapsulation with synergistic effects would efficiently provide hosts with various biological benefits and minimize the difficulty in material fabrication, inspiring next-generation material design for tackling complicated pathological conditions.

Molecular Cloning and Expression Analysis of Thyrotropin-Releasing Hormone, and Its Possible Role in Gonadal Differentiation in Rice Field eel Monopterus albus.

Simple SummaryThyrotropin-releasing hormone (TRH) is an important upstream regulator in the hypothalamus-pituitary-thyroid (HPT) axis in mammals. In this study, we isolated and characterized trh gene from a protogynous hermaphrodite fish rice field eel Monopterus albus. TRH had no significant effect on serum thyroid hormone levels in rice field eel. However, we found that TRH was involved in the regulation gonadal differentiation-related gene expression and serum sex steroid hormone secretion. Our results indicated that TRH may play a novel role in gonadal differentiation in rice field eel.Rice field eel (Monopterus albus), a protogynous hermaphrodite fish, is a good model for the research of sex determination and gonadal differentiation in teleosts. In this study, we cloned the full-length cDNA sequence of trh, which encoded a predicted protein with 270 amino acids. Trh mainly expressed in the brain, followed by the ovary, testis, muscle and pituitary, and had low levels in other peripheral tissues. During natural sex reversal, trh mRNA expression levels exhibited a significant increase at the late intersexual stage in the hypothalamus. In the gonad, trh mRNA expression levels showed a trend of increase followed by decrease, and only increased significantly at the middle intersexual stage. No matter static incubation or intraperitoneal (IP) injection, TRH had no significant effect on trh and thyroid-stimulating hormone β subunit (tshβ) mRNA expression levels, and serum T3, T4 and TRH release. After static incubation of ovarian fragments by TRH, the expression of gonadal soma derived factor (gsdf) was up-regulated significantly at both the doses of 10 and 100 nM. IP injection of TRH stimulated the expression of gsdf, and inhibited the expression of ovarian aromatase gene (cyp19a1a), accompanied by the increase of serum 11-KT levels. The results indicated that TRH may play a novel role in gonadal differentiation by the regulation of gonadal differentiation-related gene expression and sex steroid hormone secretion in rice field eel.

HPT‐ Axis Activation During Cold Head Out Water Immersion is Not Modified by Moderate Hypoxia

PurposeThe Bajau, an indigenous population renowned for breath‐hold diving, have comparatively large spleen volumes. This adaptation is purportedly indicative of a greater capacity to store red blood cells, which may aid operations in the aquatic environment. Spleen size in the Bajau is associated with the PDE10A genotype that encodes an enzyme that catalyzes the release of thyroxine (T4) from the thyroid gland. T4 is readily released during cold stress as the terminus to activation of the hypothalamus‐pituitary‐thyroid (HPT) ‐axis to increase metabolic rate. T4, and downstream counterpart triiodothyronine (T3), can also stimulate erythropoiesis by increasing erythropoietin production, which can be further promoted with hypoxia. Thus, exposure to cold water may augment erythropoiesis as hypoxia develops during breath‐hold diving via activation of the HPT‐axis. However, interactions between cold exposure and hypoxia on HPT‐axis activation have not been directly explored. Mountaineering expedition studies indirectly suggest that T4 release is elevated by hypoxia. Thus, this study tested the hypothesis that during cold head out water immersion (HOWI) acute hypoxia augments circulating thyroid hormone concentrations compared to normoxia.MethodsIn a randomized crossover single blind design, 12 healthy adults (27 ± 2 y, 2 women) completed one hour of cold (22.0 ± 0.1⁰C) HOWI breathing either normobaric normoxia (FiO2 = 0.21 ± 00, NORM) or normobaric hypoxia (FiO2 = 0.13 ± 00, HYP). Free (f) and total (t) T3 and T4, and thyroid stimulating hormone (TSH) were measured in venous blood samples obtained before (baseline), during (15‐, 30‐, and 60‐min), and 15 min following immersion (post‐). Arterial oxyhemoglobin saturation (SpO2) and rectal temperature were measured continuously. Data are presented as mean ± SD.ResultsSpO2 was lower in HYP (90 ± 3%) compared to NORM (98 ± 1%, p <.001). Rectal temperature did not differ between HYP and NORM (p > .711) but decreased over time in both trials (HYP ‐ baseline: 37.2 ± 0.4⁰C, post‐: 36.4 ± 0.5⁰C, p<.001, NORM ‐ baseline: 37.2 ± 0.4⁰C, post‐: 36.3 ± 0.5⁰C, p <.001). From baseline to post‐HOWI, fT3 (HYP: +0.3 ± 0.4 pg/mL, NORM: +0.4 ± 0.6 pg/mL, p <.001), fT4 (HYP: +1.6 ± 1.7 pg/mL, NORM: +0.7 ± 1.3 pg/mL, p <.001), tT3 (HYP: +0.3 ± 0.5 ng/mL, NORM: +0.2 ± 0.4 ng/mL, p= .053), and tT4 (HYP: +0.6 ± 0.4 µg%, NORM: +0.4 ± 0.5 µg%, p < .001) increased, but the magnitude did not differ between inspirates (p > .165). Relative to baseline, tT4 at 15 min (HYP: ‐0.2 ± 0.3 µg%, NORM: ‐0.3 ± 0.3 µg%, p = .025), and TSH at 30 min (HYP: ‐0.2 ± 0.2 µIU/mL, NORM: ‐0.2 ± 0.3 µIU/mL, p = .029) and 60 min (HYP: ‐0.2 ± 0.2 µIU/mL, NORM: ‐0.3 ± 0.4 µIU/mL, p = .008) of HOWI were lower, but did not differ between inspirates (p = .921).ConclusionsIncreases in circulating thyroid hormone concentrations during one hour of cold HOWI are not modified by normobaric hypoxia. Thus, moderate hypoxia does not likely affect activation of the HPT‐axis during short duration cold exposure. Further investigation is warranted to determine the possibility of circulating thyroid hormones to augment the erythropoietic response to acute hypoxia during cold exposure.

Screening for drugs potentially interfering with MCT8-mediated T3 transport in vitro identifies dexamethasone and some commonly used drugs as inhibitors of MCT8 activity.

Monocarboxylate transporter 8 (MCT8) is the first thyroid hormone transporter that has been linked to a human disease. Besides genetic alterations other factors might impair MCT8 activity. This study aimed at investigating whether some common drugs having a structural similarity with TH and/or whose treatment is associated with thyroid function test abnormalities, or which behave as antagonists of TH action can inhibit MCT8-mediated T3 transport. [125I]T3 uptake and efflux were measured in COS-7 cells transiently transfected with hMCT8 before and after exposure to increasing concentrations of hydrocortisone, dexamethasone, prednisone, prednisolone, amiodarone, desethylamiodarone, dronedarone, buspirone, carbamazepine, valproic acid, and L-carnitine. The mode of inhibition was also determined. Dexamethasone significantly inhibited T3 uptake at 10μM; hydrocortisone reduced T3 uptake only at high concentrations, i.e. at 500 and 1000μM; prednisone and prednisolone were devoid of inhibitory potential. Amiodarone caused a reduction of T3 uptake by MCT8 only at the highest concentrations used (44% at 50μM and 68% at 100μM), and this effect was weaker than that produced by desethylamiodarone and dronedarone; buspirone resulted a potent inhibitor, reducing T3 uptake at 0.1-10μM. L-Carnitine inhibited T3 uptake only at 500mM and 1M. Kinetic experiments revealed a noncompetitive mode of inhibition for all compounds. All drugs inhibiting T3 uptake did not affect T3 release. This study shows a novel effect of some common drugs, which is inhibition of T3 transport mediated by MCT8. Specifically, dexamethasone, buspirone, desethylamiodarone, and dronedarone behave as potent inhibitors of MCT8.

T4-mediated rescue of aortic malformations in hypothyroid rats indicates maternal thyroid status can affect great vessel development

Pexacerfont is a corticotrophin-releasing factor subtype 1 receptor (CRF-1) antagonist developed for potential treatment of anxiety and stress-related disorders. In male rats, pexacerfont caused hepatic enzyme induction leading to increased thyroxine (T4) clearance. When administered to pregnant rats on gestation day 6 to 15, pexacerfont at 300 mg/kg/day (30× mean AUC in humans at 100 mg/day) produced similar effects on thyroid homeostasis with serum T4 and thyroid-stimulating hormone levels that were 0.3–0.5× and 3.3–3.7× of controls, respectively. At this dose, fetuses of pexacerfont-treated dams presented findings associated with maternal hypothyroidism including growth retardation and increased skeletal alterations. Additionally, there were unexpected great vessel malformations that were mostly derived from the 4th pharyngeal arch artery in 5 (4.3%) fetuses from 3 (15.8%) litters. The etiology was unclear whether the vascular malformations were related to insufficient thyroid hormones or another mechanism. To better understand this relationship, pregnant rats were implanted with a subcutaneous L-thyroxine pellet designed to provide a sustained release of T4 throughout organogenesis in rat embryos (GD 6 to 15; the dosing period of pexacerfont). T4 supplementation produced a near euthyroid state in pexacerfont-treated dams and completely prevented the fetal vascular malformations. These results suggest maternal T4 levels during organogenesis may have a role in great vessel morphogenesis associated with patterning and/or regression of pharyngeal arch arteries. Although previous clinical reports have speculated a potential relationship between thyroid hormone homeostasis and early cardiovascular development, this is the first report to experimentally demonstrate this relationship in great vessel morphogenesis.

Bioinspired Nanofiber Scaffold for Differentiating Bone Marrow-Derived Neural Stem Cells to Oligodendrocyte-Like Cells: Design, Fabrication, and Characterization.

BackgroundResearchers are trying to study the mechanism of neural stem cells (NSCs) differentiation to oligodendrocyte-like cells (OLCs) as well as to enhance the selective differentiation of NSCs to oligodendrocytes. However, the limitation in nerve tissue accessibility to isolate the NSCs as well as their differentiation toward oligodendrocytes is still challenging.PurposeIn the present study, a hybrid polycaprolactone (PCL)-gelatin nanofiber scaffold mimicking the native extracellular matrix and axon morphology to direct the differentiation of bone marrow-derived NSCs to OLCs was introduced.Materials and MethodsIn order to achieve a sustained release of T3, this factor was encapsulated within chitosan nanoparticles and chitosan-loaded T3 was incorporated within PCL nanofibers. Polyaniline graphene (PAG) nanocomposite was incorporated within gelatin nanofibers to endow the scaffold with conductive properties, which resemble the conductive behavior of axons. Biodegradation, water contact angle measurements, and scanning electron microscopy (SEM) observations as well as conductivity tests were used to evaluate the properties of the prepared scaffold. The concentration of PAG and T3-loaded chitosan NPs in nanofibers were optimized by examining the proliferation of cultured bone marrow-derived mesenchymal stem cells (BMSCs) on the scaffolds. The differentiation of BMSCs-derived NSCs cultured on the fabricated scaffolds into OLCs was analyzed by evaluating the expression of oligodendrocyte markers using immunofluorescence (ICC), RT-PCR and flowcytometric assays.ResultsIncorporating 2% PAG proved to have superior cell support and proliferation while guaranteeing electrical conductivity of 10.8 × 10−5 S/cm. Moreover, the scaffold containing 2% of T3-loaded chitosan NPs was considered to be the most biocompatible samples. Result of ICC, RT-PCR and flow cytometry showed high expression of O4, Olig2, platelet-derived growth factor receptor-alpha (PDGFR-α), O1, myelin/oligodendrocyte glycoprotein (MOG) and myelin basic protein (MBP) high expressed but low expression of glial fibrillary acidic protein (GFAP). ConclusionConsidering surface topography, biocompatibility, electrical conductivity and gene expression, the hybrid PCL/gelatin scaffold with the controlled release of T3 may be considered as a promising candidate to be used as an in vitro model to study patient-derived oligodendrocytes by isolating patient’s BMSCs in pathological conditions such as diseases or injuries. Moreover, the resulted oligodendrocytes can be used as a desirable source for transplanting in patients.

Thyroid-related hormones as potential markers of hypoxia/ischemia

This study aimed to investigate the usefulness of the thyroid-related hormones as markers of acute systemic hypoxia/ischemia to identify deaths caused by asphyxiation due to neck compression in human autopsy cases. The following deaths from pathophysiological conditions were examined: mechanical asphyxia and acute/subacute blunt head injury; acute/subacute non-head blunt injury; sharp instrument injury as the hemorrhagic shock condition; drowning as alveolar injury; burn; and death due to cardiac dysfunction. Blood samples were collected from the left and right cardiac chambers and iliac veins, and serum triiodothyronine (T3), thyroxine (T4), thyroglobulin (Tg), and thyroid-stimulating hormone (TSH) levels were measured using electrochemiluminescence immunoassays. Two types of thyroid cell lines were used to confirm independent thyroid function under the condition of hypoxia (3% O2). The human thyroid carcinoma cell line (HOTHC) cell line derived from human anaplastic thyroid carcinoma and the UD-PTC (sample of the second resection papillary thyroid carcinoma) cell line derived from human thyroid papillary adenoma, which forms Tg retention follicles, were used to examine the secretion levels of T3, T4, and Tg hormones. The results showed a strong correlation between T3 and T4 levels in all blood sampling sites, while the TSH and Tg levels were not correlated with the other markers. Serum T3 and T4 levels were higher in cases of mechanical asphyxia and acute/subacute blunt head injury, representing hypoxic and ischemic conditions of the brain as compared to those in other causes of death. In the thyroid gland cell line, T4, T3, and Tg levels were stimulated after exposure to hypoxia for 10–30 min. These findings suggest that systemic advanced hypoxia/ischemia may cause a rapid and TSH-independent release of T3 and T4 thyroid hormones in autopsy cases. These findings demonstrate that increased thyroid-related hormone (T3 and T4) levels in the pathophysiological field may indicate systemic hypoxia/ischemia.

Protective effect of a polyherbal bioactive fraction in propylthiouracil-induced thyroid toxicity in ratsby modulation of the hypothalamic–pituitary–thyroid and hypothalamic–pituitary–adrenal axes

Hypothyroidism is the most frequent consequence of the interaction of a large variety of drugs, environmental pollutants and industrial chemicals with the thyroid gland. It is associated with diminished endocrine function which may lead to hyperlipidemia, diabetes, Alzheimer’s disease, weight gain, and other metabolic disorders. The present study evaluates the pro-thyroid activity of a bioactive fraction from a polyherbal teabag in rats with hypothyroidism induced by propylthiouracil. The teabag was formulated to stimulate synthesis and/or release of T4 and affectthe conversion of T4 to T3. Phytoconstituents of the polyherbal teabag are potent antioxidants that may be responsible for the pro-thyroid activity. The tea-extract (1000 mg) was found to contain 1076 μg of gallic acid and 1131 μg of rutin from HPTLC analysis. Rats received propylthiouracil (8 mg/kg) for the first 15days followed by the polyherbal tea-extract (500, 1000 and 1500 mg/kg), the standard drug levothyroxine (0.1 mg/kg), aerobic exercise, and a combination of tea-extract (1000 mg/kg) and aerobic exercise daily along with propylthiouracil for the next 30 days. Finally, rats received their respective treatments alone without propylthiouracil for 15 more days. Lipid profile and levels of glucose, insulin, T3, T4, TSH, cortisol, homocysteine, creatinine, uric acid, malondialdehyde, glucose-6 phosphatase, and endogenous antioxidants were determined. All treatments attenuated significantly the propylthiouracil-elevated TSH, homocysteine, creatinine, uric acid, glucose-6-phosphatase, insulin, and malondialdehyde levels, and restored favorably the propylthiouracil-altered lipid profile, T3, T4, and endogenous antioxidant levels. The polyherbal tea-extract (1000 and 1500 mg/kg) treatment and thecombination treatment of tea-extract (1000 mg/kg) with aerobic exercise displayed significant restoration of the suboptimalthyroid function. This may be due to a favorablemodulation ofthe hypothalamic-pituitary–thyroid and the hypothalamic–pituitary–adrenal axes.

Polychlorinated biphenyl toxicity in the thyroid gland of wild ungulates: an in vitro model

Polychlorinated biphenyls (PCBs) are carcinogens causing endocrine disruption. While production of PCBs is now banned, wildlife exposure still occurs due to environmental contamination. We investigated thyroid toxicity in wild ungulates using three-dimensional primary thyrocyte cultures exposed to PCB 138 for 24, 48, and 72 h at concentrations ranging within 0–3000 ng/ml. Thyrocyte viability ranged within 78.71–118.34%, 98.14–104.45%, and 84.16–106.70% in fallow deer-, mouflon-, and roe deer-derived cells, respectively. Viability decreased significantly in fallow deer (P = 0.012) and roe deer (P = 0.002) thyrocytes exposed for 48 h at 30 ng/ml. While cytotoxicity ranged within 2.36–16.37%, 3.19–9.85%, and 2.76–11.21% in fallow deer, mouflon, and roe deer, respectively, only roe deer displayed significantly higher cytotoxicity at a 3 ng/ml exposure (P < 0.05) and lower cytotoxicity at 30 ng/ml (P < 0.01). Exposure to 30 ng/ml for 24 and 48 h induced reactive oxygen species in fallow deer. Iodide uptake at 30 ng/ml exposure increased after 24 h in fallow and roe deer, but showed a significant drop after 48 and 72 h in fallow deer, mouflon, and roe deer. Thyroxine T4 release at 30 ng/ml exposure decreased significantly after 48 and 72 h; 24, 48 and 72 h; and 48 h in fallow deer, mouflon, and roe deer, respectively. Our findings indicate time- and species-dependent effects of PCB on performance and thyrocyte function. Use of cell culture models reduces the number of experimental specimens, increases test species welfare and replaces whole organisms with specific target cells.

A CASE STUDY ON HYPOTHYROIDISM IN CHHATTISGARH

Hypothyroidism in Chhattisgarh (India) is almost the second most common disease in middle aged and old aged females. First may be Diabetes / Hypertension / Arthritis / Osteoporosis. Hypothyroidism is caused due to under functioning Thyroid gland may be due to primary or secondary causes. The Etiology is still a mystery & is unknown but having strong relation with genetics as well as the environmental pollution, food habits & autoimmunity. The Thyroid gland regulates the metabolism of the body through the release of T3, T4 hormones & have a negative feedback of TSH hormone. A case study was conducted on 70 patients from “Sai Kripa Clinic” Main Road Katora Talab Raipur , Chattisgarh, India.
 Key words: Hypothyroidism, Autoimmune Disease, Obesity, Arthritis.

Endocrine disruption as an adverse effect of non-endocrine targeting pharmaceuticals.

Endocrine disruptors have gained widespread attention owing to their severe adverse health impacts. These produce enormous burden of disease and are associated with high economic cost especially in developed countries. Environmental pollutants causing endocrine disruption include pesticides, industrial wastes, packaging materials, food constituents, plastics, and cosmetic products. Likewise, pharmaceutical drugs have the endocrine disrupting potential through a wide array of mechanisms. Antipsychotic, antiepileptic, antihypertensive, antiviral, antidiabetic, and anticancer drugs are among the foremost non-hormonal endocrine disruptors. Several drugs affect thyroid hormone synthesis via interaction with iodine uptake to the release of T3 and T4 by thyrocytes. Prolonged use of some drugs increase susceptibility to diabetes mellitus either by direct destruction of β cells or enhanced insulin resistance. Other drugs may cause serious developmental defects in male or female reproductive system. Appropriate understanding of the mechanisms of endocrine disruption associated with non-hormonal drugs will guide future drug development and help us prevent and cure endocrine-related toxicity of pharmaceuticals. Therefore, this review focuses on endocrine disruption by pharmaceutical drugs as their side effect.

Metal Coordinated Poly-Zinc-Liothyronine Provides Stable Circulating Triiodothyronine Levels in Hypothyroid Rats.

Liothyronine (LT3) has limited short-term clinical applications, all of which aim at suppressing thyrotropin (TSH) secretion. A more controversial application is chronic administration along with levothyroxine in the treatment of hypothyroidism. Long-term treatment with LT3 is complicated by its unique pharmacokinetics that result in a substantial triiodothyronine (T3) peak in the blood three to four hours after oral dosing. This is a significant problem, given that T3 levels in the blood are normally stable, varying by <10% throughout the day. A metal coordinated form of LT3 (Zn[T3][H2O])n, known as poly-zinc-liothyronine (PZL), was synthesized and loaded into coated gelatin capsules for delivery to the duodenum where sustained release of T3 from PZL occurs. Male Wistar rats were made hypothyroid by feeding on a low iodine diet and water containing 0.05% methimazole for five to six weeks. Rats were given a capsule containing 24 μg/kg PZL or equimolar amounts of LT3. Blood samples were obtained multiple times from the tail vein during the first 16 hours, and processed for T3 and TSH serum levels. Some animals were treated daily for eight days, and blood samples were collected daily. Rats given LT3 exhibited the expected serum T3 peak (about fivefold baseline) at 3.5 hours, followed by a rapid decline, with serum levels almost returning to baseline values by 16 hours. In contrast, serum T3 in PZL-treated rats exhibited about a 30% lower T3 peak at nine hours. Furthermore, the plateau time, that is, the time-span during which the serum T3 concentration is at least half of T3 peak, increased from 4.9 to 7.7 hours in LT3- versus PZL-treated rats, respectively. Serum TSH dropped in both groups, but PZL-treated rats exhibited a more gradual decrease, which was delayed by about four hours compared to LT3-treated rats. Chronic treatment with either LT3 or PZL restored growth, lowered serum cholesterol, and stimulated hepatic expression of the Dio1 mRNA and other T3-dependent markers in the central nervous system. Capsules of PZL given orally restore T3-dependent biological effects while exhibiting a reduced and delayed serum T3 peak after dosing, thus providing a longer period of relatively stable serum T3 levels compared to capsules of LT3.

Sex Differences in Brain Thyroid Hormone Levels during Early Post-Hatching Development in Zebra Finch (Taeniopygia guttata).

Thyroid hormones are closely linked to the hatching process in precocial birds. Previously, we showed that thyroid hormones in brain had a strong impact on filial imprinting, an early learning behavior in newly hatched chicks; brain 3,5,3′-triiodothyronine (T3) peaks around hatching and imprinting training induces additional T3 release, thus, extending the sensitive period for imprinting and enabling subsequent other learning. On the other hand, blood thyroid hormone levels have been reported to increase gradually after hatching in altricial species, but it remains unknown how the brain thyroid hormone levels change during post-hatching development of altricial birds. Here, we determined the changes in serum and brain thyroid hormone levels of a passerine songbird species, the zebra finch using radioimmunoassay. In the serum, we found a gradual increase in thyroid hormone levels during post-hatching development, as well as differences between male and female finches. In the brain, there was clear surge in the hormone levels during development in males and females coinciding with the time of fledging, but the onset of the surge of thyroxine (T4) in males preceded that of females, whereas the onset of the surge of T3 in males succeeded that of females. These findings provide a basis for understanding the functions of thyroid hormones during early development and learning in altricial birds.

Inhaling Benzene, Toluene, Nitrogen Dioxide, and Sulfur Dioxide, Disrupts Thyroid Function in Captive American Kestrels (Falco sparverius).

Research investigating the effects of air contaminants on biota has been limited to date. Captive adult female American kestrels (Falco sparverius) were exposed to a mixture of benzene (0.6 ppm), toluene (1 ppm), nitrogen dioxide (NO2; 2 ppm) and sulfur dioxide (SO2; 5.6 ppm), in a whole-body inhalation chamber. Thyroid axis responses to meet metabolic demands were examined through thyroid histology, plasma thyroxine (T4), and triiodothyronine (T3), and hepatic outer ring deiodination (T4-ORD). Plasma free (F) T3 and T4 were measured at baseline, and at 9 days and 18 days of exposure, whereas total (T) T3 and TT4, thyroid histology and hepatic T4-ORD were determined at the final 18 day exposure. Inhalation of these contaminants significantly suppressed plasma FT4 and TT4, and depleted follicular colloid and increased epithelial cell height at 18 days, and significantly altered the temporal pattern of plasma FT4. Significant histological changes in the follicular colloid:epithelial cell height ratio indicated sustained T4 production and release by the thyroid glands. There was no effect on plasma FT3, TT3, or hepatic T4-ORD. We hypothesize that contaminant-related activation of the hypothalamus-pituitary-thyroid axis in the kestrels increased elimination of plasma T4 through Phase II enzymes. Further research is required to test this hypothesis in wild birds.

A model for chronic, intrahypothalamic thyroid hormone administration in rats.

In addition to the direct effects of thyroid hormone (TH) on peripheral organs, recent work showed metabolic effects of TH on the liver and brown adipose tissue via neural pathways originating in the hypothalamic paraventricular and ventromedial nucleus (PVN and VMH). So far, these experiments focused on short-term administration of TH. The aim of this study is to develop a technique for chronic and nucleus-specific intrahypothalamic administration of the biologically active TH tri-iodothyronine (T3). We used beeswax pellets loaded with an amount of T3 based on in vitro experiments showing stable T3 release (∼5 nmol l(-1)) for 32 days. Upon stereotactic bilateral implantation, T3 concentrations were increased 90-fold in the PVN region and 50-fold in the VMH region after placing T3-containing pellets in the rat PVN or VMH for 28 days respectively. Increased local T3 concentrations were reflected by selectively increased mRNA expression of the T3-responsive genes Dio3 and Hr in the PVN or in the VMH. After placement of T3-containing pellets in the PVN, Tshb mRNA was significantly decreased in the pituitary, without altered Trh mRNA in the PVN region. Plasma T3 and T4 concentrations decreased without altered plasma TSH. We observed no changes in pituitary Tshb mRNA, plasma TSH, or plasma TH in rats after placement of T3-containing pellets in the VMH. We developed a method to selectively and chronically deliver T3 to specific hypothalamic nuclei. This will enable future studies on the chronic effects of intrahypothalamic T3 on energy metabolism via the PVN or VMH.

Thyroid gland in vitro activity and reactivity to TRH in newborn lambs

The aim of the study was to determine the thyroid gland tissue concentration and activity on T4 and T3 in vitro release to the medium without (control) or with TRH (10μg/ml) for 90min of incubation in lambs just after birth and after the first 6h of postnatal life.The results obtained show that T4 concentration in the blood plasma in newborn lambs as well after 6h of life is higher than in thyroid gland tissue. In the case of T3 the concentration in blood plasma of lambs just after birth is lower than after 6h of postnatal life. Inversely, the higher level of that hormone concentration in blood plasma just after birth is higher than in the thyroid gland tissue. The results clearly indicate the inhibitory TRH effect on in vitro T4 especially, and in lesser degree T3 release from the thyroid gland tissue of lambs just after birth. Stimulation of the TRH thyroid gland tissue from lambs after 6h of life caused an increase in T4 release, especially after 60min of the experiment. Similarly, a significant increase of T3 release was found only after 60min of incubation. The transient promoting TRH effect on the amount of iodothyronine release from the thyroid gland in older lambs is probably connected with this gland's function not occurring before the designated time of postnatal adaptation. The described in vitro experiment visibly shows that changing thyroid gland activity in newborn lambs is not only conditioned by the entire HPT activity, but by the immanent thyroid gland attributes as well.