The effect of vitamin A supplementation on thyroid function in premenopausal women.

OBJECTIVE: Vitamin A and its retinoid derivates play an important role in regulation of normal growth and development. Vitamin A has been shown to regulate thyroid hormone metabolism and inhibit thyroid-stimulating hormone (TSH) secretion via down regulation of TSH-ß gene expression; however, the effect of vitamin A on thyroid function in obese individuals who are at higher risk of subclinical hypothyroidism is still unclear. In the present study we investigate the impact of vitamin A supplementation on thyroid function in obese women. METHOD: A 4-month randomized, double blind controlled trial was conducted among 84 healthy women aged 17-50 years old: 56 were obese (body mass index [BMI] 30-35 kg/m(2)) and 28 were nonobese (BMI 18.5-24.9 kg/m(2)). Obese women were randomly allocated to receive either vitamin A (25,000 IU/d retinyl palmitate) or placebo. Nonobese women received vitamin A. At baseline and 4 months after intervention, serum concentrations of TSH, total thyroxine (T4), total triiodothyronine (T3), retinol-binding protein (RBP), and transthyretin (TTR) were measured. RESULTS: Baseline concentrations of thyroid hormones, RBP and TTR were not significantly different between groups. Vitamin A caused a significant reduction in serum TSH concentrations in obese (p = 0.004) and nonobese (p = 0.001) groups. Serum T3 concentrations also increased in both obese and nonobese vitamin A-treated groups (p < 0.001). Serum T4 decreased in all 3 groups after treatment. The results showed a significant reduction in serum RBP in the obese group after vitamin A supplementation (p = 0.007), but no significant change was seen in serum TTR. CONCLUSIONS: Serum TSH concentrations in vitamin A-treated subjects were significantly reduced; therefore, vitamin A supplementation might reduce the risk of subclinical hypothyroidism in premenopausal women.

Increasing plasma thyroxine levels during late embryogenesis and hatching in the chicken are not caused by an increased sensitivity of the thyrotropes to hypothalamic stimulation.

The hatching process in the chicken is accompanied by dramatic changes in plasma thyroid hormones. The cause of these changes, though crucial for hatching and the onset of endothermy, is not known. One hypothesis is that the pituitary gland becomes more sensitive to hypothalamic stimulation during this period. We have tested whether the responsiveness of the thyrotropes to hypothalamic stimuli changes throughout the last week of embryonic development and hatching by studying the mRNA expression of receptors involved in the control of the secretory activity of this cell type. We used a real-time PCR set-up to quantify whole pituitary mRNA expression of the beta subunit of thyrotrophin (TSH-beta), type 1 thyrotrophin-releasing hormone receptor (TRH-R1), corticotrophin-releasing hormone receptors (CRH-R1 and CRH-R2) and somatostatin subtype receptor 2 (SSTR2) on every day of the last week of embryonic development, including the day of hatch and the first day of posthatch life. The thyrotrope-specific expression was investigated by a combination of in situ hybridization with immunohistochemistry at selected ages. Although TSH-beta mRNA levels increased towards day 19 of incubation (E19), the expression of CRH-R2 and TRH-R1 mRNA by the thyrotropes tended to decrease during this period, suggesting a lower sensitivity of the thyrotropes to the stimulatory factors CRH and TRH. CRH-R1, which is not involved in the control of TSH secretion, increased steadily throughout the period tested. The expression of SSTR2 mRNA by the thyrotropes was low during embryonic development and increased just before hatching. We have concluded that the sensitivity of the pituitary thyrotropes to hypothalamic stimulation decreases throughout the last week of embryonic development, so that the higher expression of TSH-beta mRNA around E16-E19, and hence the increasing plasma thyroxine level, is unlikely to be the result of an increased stimulation by either TRH or CRH.

Ontogeny of pituitary thyrotrophs and regulation by endogenous thyroid hormone feedback in the chick embryo.

Increased thyroid hormone production is essential for hatching of the chick and for the increased metabolism necessary for posthatch endothermic life. However, little is known about the ontogeny and distribution of pituitary thyrotrophs during this period or whether pituitary thyroid-stimulating hormone (TSH) production is regulated by endogenous thyroid hormones during chick embryonic development. This study assessed the abundance and location of pituitary thyrotrophs and the regulation of TSH(beta) peptide and mRNA levels by endogenous thyroid hormones prior to hatching. TSH(beta)-containing cells were first detected on embryonic day (e) 11, and the thyrotroph population increased to maximum levels on e17 and e19 and then decreased prior to hatching (d1). Thyrotroph distribution within the cephalic lobe of the anterior pituitary was determined on e19 by whole-mount immunocytochemistry for TSH(beta) peptide and by whole-mount in situ hybridization for TSH(beta) mRNA. Thyrotroph distribution within the cephalic lobe was heterogeneous among embryos, but most commonly extended from the ventral medial region to the dorsal lateral regions, along the boundary of the cephalic and caudal lobes. Inhibition of endogenous thyroid hormone production with methimazole (MMI) decreased plasma thyroxine (T4) levels and increased pituitary TSH(beta) mRNA levels on e19 and d1. However, control pituitaries contained significantly more TSH(beta) peptide than MMI-treated pituitaries on e17 and e19, suggesting higher TSH secretion into the blood in MMI-treated groups. We conclude that thyrotroph abundance and TSH production increase prior to hatching, that thyrotrophs are localized heterogeneously within the cephalic lobe of the anterior pituitary at that time, and that TSH gene expression and secretion are under negative feedback regulation from thyroid hormones during this critical period of development.

Cushing's disease after surgical resection and radiation therapy for nonfunctioning pituitary adenoma.

OBJECTIVE: To describe a patient with an aggressive nonfunctioning pituitary adenoma in whom Cushing's disease developed after two resections of tumor and radiation therapy. METHODS: We present a case report, with serial laboratory and immunohistochemical data, and summarize information about similar patients described in the medical literature. RESULTS: A 48-year-old woman had irregular menstrual periods, decreased peripheral vision, headaches, and weight gain. Laboratory and radiologic investigation revealed a large nonfunctioning pituitary adenoma. Transsphenoidal subtotal resection of the tumor improved her vision. Results of immunohistochemical studies were positive for b-follicle-stimulating hormone and b-luteinizing hormone. She had radiation therapy 1 year postoperatively for rapidly enlarging residual tumor. Bifrontal craniotomy was done 3 months later because of worsening vision. The pituitary adenoma from the second surgical procedure stained negatively for all pituitary hormones. Postoperatively, she received tapering doses of prednisone for 4 months. Two months after the last dose of prednisone, she had signs and symptoms of hypercortisolism. Inferior petrosal sinus venous sampling studies for plasma corticotropin confirmed the presence of Cushing's disease. She did not tolerate medical therapy. Bilateral adrenalectomy led to remission of hypercortisolism. CONCLUSION: Nonfunctioning pituitary tumors often come to clinical attention when they are large and cause symptoms associated with hypopituitarism or invasion of parasellar structures. In contrast, functioning pituitary tumors may have few compressive symptoms if they manifest with complaints attributable to excessive pituitary hormones.

Two novel mutations of the TSH-beta subunit gene underlying congenital central hypothyroidism undetectable in neonatal TSH screening.

CONTEXT: Patients with TSH-beta subunit defects and congenital hypothyroidism are missed by TSH-based neonatal screening. OBJECTIVE: Our objective was to report the molecular consequences of a novel splice-junction mutation and a novel missense mutation in the TSH-beta subunit gene found in two patients with congenital central hypothyroidism and conventional treatment-resistant anemia. RESULTS: Patient 1 had a homozygous G to A nucleotide change at the 5' donor splice site of exon/intron 2. This resulted in a silent change at codon 34 of the mature protein. In vitro splicing assays showed that the mutant minigene dramatically affected pre-mRNA processing, causing exon 2 to be completely skipped. The putative product from a new out-of-frame translational start point in exon 3 is expected to yield a nonsense 25-amino-acid peptide. In patient 2, sequence analysis revealed a compound heterozygosis for the already reported 313delT (C105Vfs114X) mutation and for a second novel mutation in exon 3, substituting G for A at cDNA nucleotide position 323, resulting in a C88Y change. This cysteine residue is conserved among all dimeric pituitary and placental glycoprotein hormone-beta subunits. Data from in silico analysis confirmed that the C88Y mutation would affect subunit conformation. Indeed, two different bioinformatics approaches, PolyPhen and SIFT analysis, predicted C88Y to be a damaging substitution. CONCLUSIONS: In isolated TSH deficiency, the exact molecular diagnosis is mandatory for diagnosis of isolated pituitary deficiency, delineation of prognosis, and genetic counseling. Moreover, diagnosis of central hypothyroidism should be considered in the face of severe infant anemia of uncertain etiology.

Expression and hypophysiotropic actions of corticotropin-releasing factor in Xenopus laevis.

Members of the corticotropin-releasing factor (CRF) family of peptides play pivotal roles in the regulation of neuroendocrine, autonomic, and behavioral responses to physical and emotional stress. In amphibian tadpoles, CRF-like peptides stimulate both thyroid and interrenal (adrenal) hormone secretion, and can thereby modulate the rate of metamorphosis. To better understand the regulation of expression and actions of CRF in amphibians we developed a homologous radioimmunoassay (RIA) for Xenopus laevis CRF (xCRF). We validated this RIA and tissue extraction procedure for the measurement of brain CRF content in tadpoles and juveniles. We show that the CRF-binding protein, which is highly expressed in X. laevis brain, is largely removed by acid extraction and does not interfere in the RIA. We analyzed CRF peptide content in five microdissected brain regions in prometamorphic tadpoles and juveniles. CRF was detected throughout the brain, consistent with its role as both a hypophysiotropin and a neurotransmitter/neuromodulator. CRF content was highest in the region of the preoptic area (POa) and increased in all brain regions after metamorphosis. Exposure to 4h of handling/shaking stress resulted in increased CRF peptide content in the POa in juvenile frogs. Injections of xCRF into prometamorphic tadpoles increased whole body corticosterone and thyroxine content, thus supporting findings in other anuran species that this peptide functions as both a corticotropin- and a thyrotropin (TSH)-releasing factor. Furthermore, treatment of cultured tadpole pituitaries with xCRF (100nM for 24h) resulted in increased medium content, but decreased pituitary content of TSHbeta-immunoreactivity. Our results support the view that CRF functions as a stress neuropeptide in X. laevis as in other vertebrates. Furthermore, we provide evidence for a dual hypophysiotropic action of CRF on the thyroid and interrenal axes in X. laevis as has been shown previously in other amphibian species.

Utilidad de la determinación de la subunidad alfa tras estímulo con TRH como indicador de persistencia y/o recurrencia tumoral en gonadotropinomas / Utility of alpha subunit determination alter thyrotropin-releasing hormone stimulation as an indicator of gonadotropinoma persistente and/or recurrence

Objetivo: Los gonadotropinomas son tumores originados en las células gonadotropas de la hipófisis anterior causales de la síntesis y la secreción de gonadotrofinas (folitropina [FSH] y lutropina [LH]). La mayoría de estos tumores tienen una producción alterada de gonadotropinas y de sus subunidades (folitropina beta, subunidad alfa y, con menos frecuencia, lutropina beta). Los gonadotropinomas pueden presentar una respuesta de la subunidad alfa de las gonadotropinas al estímulo con protirrelina (TRH) que podría diferenciar estos tumores de los no funcionantes. De igual forma, esta prueba podría ser de utilidad tras la cirugía para poderdiscernir los posibles restos tumorales respecto a los cambios posquirúrgicos. Sujetos y método: Se estudió a 24 pacientes intervenidos de macroadenoma hipofisario, de los que 14 fueron diagnosticados de gonadotropinoma en el estudio histológico. Se les practicó la prueba de lasubunidad alfa tras la administración de TRH antes y después de la cirugía.Resultados: En el estudio prequirúrgico el 50% de los gonadotropinomas tuvieron una respuesta positiva a dicha prueba y en el posquirúrgico otro 50%. El 83% de los pacientes con gonadotropinoma presentaban signos de recidiva/persistencia tumoral y/o cambios en laresonancia magnética (RM) de control posquirúrgico; el 83% de estos pacientes (41,6% del total) tuvo una respuesta positiva de la subunidad alfa tras el estímulo con TRH. En el grupo de macroadenomas no gonadotropinomas sólo un 33% tuvo respuesta positiva antes de la cirugía y otro 33%, después. En la RM practicada después de la cirugía, todos mostraban signos radiológicos compatibles con cambios inflamatorios posquirúrgicos o signos de persistencia y/o recidivatumoral. Conclusiones: Dicha prueba podría ser de ayuda en el diagnóstico diferencial de los gonadotropinomas, así como en el seguimiento y la valoración posquirúrgica de estos tumores (AU)

Objective: Gonadotropinomas are adenomas of the gonadotropic cells of the anterior pituitary. These cells produce and secrete gonadotropins (follicle-stimulating hormone and luteinizing hormone). Most of these tumors show altered production of gonadotropins and their subunits (the - FSH, and, less frequently, -LH subunits). The thyrotropin-releasing hormone (TRH) stimulation test could differentiate these tumors from nonfunctioning tumors. Equally, this test could be able to distinguish between postsurgical changes and tumoral remnants after surgery. Subjects and method: We studied 24 patients with pituitary macroadenoma, 14 of who had a histological diagnosis of gonadotroph adenoma. The TRH stimulation test was performed before and after surgery. Results: Both before and after surgery, a positive result to the TRH test was obtained in 50% of gonadotropinomas. Magnetic resonance imaging (MRI)performed after surgery revealed that 83% of the patients with gonadotropinoma had signs of tumoral persistence or recurrence and/or postsurgical changes. Of these patients, 83% (41.6% of the total) showed positive (..) (AU)