Synergic actions of polyphenols and cyanogens of peanut seed coat (Arachis hypogaea) on cytological, biochemical and functional changes in thyroid.

In animals, long-term feeding with peanut (Arachis hypogaea) seed coats causes hypertrophy and hyperplasia of the thyroid gland. However, to date there have been no detailed studies. Here, we explored the thyroidal effects of dietary peanut seed coats (PSC) in rats. The PSC has high levels of pro-goitrogenic substances including phenolic and other cyanogenic constituents. The PSC was mixed with a standard diet and fed to rats for 30 and 60 days, respectively. Animals fed with the PSC-supplemented diet showed a significant increase in urinary excretion of thiocyanate and iodine, thyroid enlargement, and hypertrophy and/or hyperplasia of thyroid follicles. In addition, there was inhibition of thyroid peroxidase (TPO) activity, 5’-deiodinase-I (DIO1) activity, and (Na+-K+)-ATPase activity in the experimental groups of rats as compared to controls. Furthermore, the PSC fed animals exhibited decreased serum circulating total T4 and T3 levels, severe in the group treated for longer duration. These data indicate that PSC could be a novel disruptor of thyroid function, due to synergistic actions of phenolic as well as cyanogenic constituents.

Effects of magnesium on cytomorphology and enzyme activities in thyroid of rats.

Till date knowledge regarding the effects of high dietary magnesium on thyroid gland is incomprehensive though certain epidemiological studies reported development of thyroid gland dysfunctions in people with chronic exposure to hard water (especially with high magnesium) despite sufficient iodine consumption. The present study is to explore the effects of chronic high dietary magnesium exposure on thyroid morphology and functional status. Male adult albino Wistar strain rats were treated with graded doses of magnesium sulphate (MgSO4; 0.5, 1.0 and 1.5 g %) for 60 days and changes in different thyroid parameters were investigated. Significantly stimulated thyroid peroxidase and Na+–K+-ATPase and altered idothyronine 5/- deiodinase type I activities, enhanced serum thyroxine (T4) (both total and free), total triiodothyronine (T3) and thyroid stimulating hormone with decreased free T3 levels and T3/T4 ratio (T3:T4) along with enlargement of thyroid with associated histopathological changes were observed in the treated groups. The results clearly confirm that chronic high dietary magnesium exposure causes potential thyroid disruption as reported in earlier epidemiological studies.

Evaluation of the role of ultrasonography in diagnosis of autoimmune thyroiditis in goitrous children.

OBJECTIVES: To evaluate the role of thyroid ultrasound in children with autoimmune thyroiditis diagnosed either on cytopathology or by the presence of thyroid peroxidase antibodies. DESIGN AND SETTINGS: Children presenting for the first time to the Thyroid Clinic at the Institute of Nuclear Medicine and Allied Sciences for the complaint of goiter over a two year period (January 2005-December 2006) were studied. SUBJECTS: 695 school children (244 boys and 451 girls) aged 5-18 year were studied. METHODS: Children were subjected to thyroid ultrasound, cytopathology, thyroid peroxidase antibody and thyroid function tests. RESULTS: Overall, 16% of goitrous children had hypoechogenicity on ultrasound, 15.2% had cytopathological evidence of thyroiditis, 10.6% had positive thyroid peroxidase antibodies and 25.2% had abnormal thyroid function tests. Subjects with hypoechogenicity had higher percentage of thyroiditis on cytopathology (41.4% vs. 10.3%; P<0.01), thyroid peroxidase antibody positivity (30.6% vs. 6.8%; P<0.01) and thyroid dysfunction (46.8% vs. 21.2%; P<0.01) than those with normal echogenicity. CONCLUSION: Thyroid USG has a useful, though limited, role in excluding thyroid disease in children. The sensitivity of echogenicity for the diagnosis of autoimmune thyroiditis in children is less than that reported in adults.

Expressão das iodotironinas desiodases nas neoplasias tireoidianas / Iodothyronine deiodinases expression in thyroid neoplasias

As iodotironinas desiodases formam uma família de selenoenzimas com propriedades catalíticas distintas que ativam ou inativam os hormônios tireoidianos via desiodação do anel fenólico ou tirosínico da molécula do T4. As desiodases tipo I e II (D1 e D2) são as enzimas responsáveis pela geração do T3 e são amplamente expressas na tireóide normal. A transformação neoplásica benigna ou maligna da glândula tireóide está associada a alterações na expressão dessas isoenzimas, sugerindo um possível papel da D1 e da D2 como marcadores de diferenciação celular. Anormalidades na expressão de ambas enzimas e da desiodase tipo III (D3), inativadora do hormônios tireoidianos, são também encontradas em outras neoplasias humanas. Os mecanismos ou implicações do aumento ou diminuição das desiodases na patogênese neoplásica são pouco compreendidas. No entanto, é importante observar que a expressão anormal da D2 pode ser responsável por um quadro de tireotoxicose em pacientes com metástases de carcinoma folicular de tireóide, enquanto que o aumento da D3 em hemangiomas pode causar hipotireoidismo de difícil tratamento.

The iodothyronine deiodinases constitute a family of selenoenzymes that catalyze the removal of iodine from the outer ring or inner ring of the thyroid hormones. The activating enzymes, deiodinases type I (D1) and type II (D2), are highly expressed in normal thyroid gland. Benign or malignant neoplastic transformation of the thyroid cells is associated with changes on the expression of these enzymes, suggesting that D1 or D2 can be markers of cellular differentiation. Abnormalities on the expression of both enzymes and also of the deiodinase type III (D3), that inactivates thyroid hormones, have been found in other human neoplasias. So far, the mechanism or implications of these findings on tumor pathogenesis are not well understood. Nevertheless, its noteworthy that abnormal expression of D2 can cause thyrotoxicosis in patients with metastasis of follicular thyroid carcinoma and that increased D3 expression in large hemangiomas causes severe hypothyroidism.

Hypercholesterolemia and tissue-specific differential mRNA expression of type-1 5'-iodothyronine deiodinase under different selenium status in rats

Type-1 5'-iodothyronine deiodinase (5'-DI) is responsible for conversion of T4 to T3. Selenium (Se) is an integral part of this enzyme. Keeping in view the strong association between atherosclerosis and hypothyroidism, the present study examined the behavior of 5'-DI in liver, aorta and thyroid during hypercholesterolemia following different Se status, i.e., Se deficiency (0.02ppm), adequate (0.2ppm) and excess dose (1ppm) in SD male rats. Animals were fed a control or high-cholesterol diet (2%) for 1 and 2 months. 5'-DI activity and mRNA expression was measured by RIA and RT-PCR respectively. In liver and aorta, 5'-DI expression significantly decreased with the Se-deficient and the high-cholesterol diet. The trend was opposite in thyroid, i.e., mRNA expression increased significantly during selenium deficiency and with a high-cholesterol feeding. But with 1ppm Se supplementation, the 5'-DI expression increased in all the three tissues. The present study indicates that hypercholesterolemia along with selenium deficiency is co-responsible for differential regulation of 5'-DI enzyme in thyroidal vs. extrathyroidal tissues. Distinct regulation of 5'-DI in the thyroid reflects the clinical importance of this selenoprotein during hypercholesterolemia as this enzyme is essential for T3 production, which further has a vital role in the maintenance of lipid metabolism.

Enzimas envolvidas na organificação tireoideana do iodo / Enzymes involved in thyroid iodide organification

A biossíntese dos hormônios da tireóide depende do funcionamento normal de uma série de proteínas que são necessárias tanto para a captação de iodeto através da membrana basolateral dos tireócitos como para sua incorporação à proteína aceptora, a tireoglobulina (Tg), o que ocorre na superfície apical da célula folicular. O co-transportador sódio-iodeto (NIS) é responsável pela captação tireoideana de iodeto, a primeira etapa da biossíntese hormonal tireoideana. No pólo apical dos tireócitos, o iodeto é transportado através da membrana celular pela pendrina (PDS) e subseqüentemente incorporado à Tg, uma proteína de alto peso molecular secretada no lúmen folicular. A oxidação do iodeto e sua organificação parecem ocorrer principalmente na superfície apical da célula folicular, e estas reações são catalisadas pela tireoperoxidase (TPO) na presença de peróxido de hidrogênio. Assim, a organificação tireoideana do iodo depende da atividade TPO, a qual é modulada pelas concentrações de substrato (tireoglobulina e iodeto) e cofator (peróxido de hidrogênio). A enzima responsável pela geração de peróxido de hidrogênio associada à hormonogênese tireoideana é a NADPH oxidase (ThOx), que encontra-se no pólo apical dos tireócitos, é estimulada pela tireotrofina e inibida pelo iodo. Aparentemente, a geração de peróxido de hidrogênio é o passo limitante da biossíntese dos hormônios da tireóide em condições de suficiência de iodo.

Atividade colinesterásica em tireóide de ratos: resposta a uma sobrecarga de iodo / Cholinesterase activity in thyroid of rats: response the a iodine overload

A utilização do iodo antes de cirurgias de tireóide visa a redução do fluxo sanguíneo para a glândula e melhora da hemostasia. Para tentar elucidar o mecanismo pelo qual a vasoconstricção se instala, foi determinada a atividade colinesterásica em tireóides de ratos Wistar machos (240-250g de peso corporal), submetidos a uma sobrecarga oral de iodo pela oferta de solução aquosa de iodato de potássio (KIO 3 3µg/ml) como água de beber, por 3 e 7 dias; o grupo controle recebeu água durante o mesmo período. Ao final do tratamento os animais foram sacrificados e suas tireóides retiradas, limpas e pesadas; o rim direito e o lobo direito do fígado foram usados como controles da atividade colinesterásica. Foram preparados homogeneizados em tampão fosfato (pH=8,0), contendo 0,32M de sacarose, na concentração de 40mg de tecido/ml. Os ratos que receberam KIO 3 por 7 dias tiveram diminuição no peso da tireóide quando comparados aos controles (p<0,05), o mesmo não acontecendo aos 3 dias de tratamento. Aos 3 dias de tratamento, a atividade colinesterásica tireóidea foi de 5,38ñ0,36 nmol/min.mg para o grupo controle (n=12) e de 5,43ñ0,98 nmol/min.mg para o grupo tratado (n=11) (NS); valores bastante similares foram também encontrados com o tratamento de 7 dias (controles: 5,42ñ0,27 nmol/min.mg [n=8] e tratados: 5,63ñ0,88 nmol/min.mg [n=8]; NS). Nossos resultados sugerem que, a despeito da vasoconstricção, não há alteração da atividade colinesterásica na tireóide em resposta a sobrecarga oral de iodo.

Sistema enzimático gerador de peróxido de hidrogênio: NADPH oxidase na tireóide humana / Enzimatic system generator of hydrogen peroxide: NADPH oxidase in human thyroid

No presente estudo avaliamos a atividade geradora de peróxido de hidrogênio (H 2 O 2 ) em frações particuladas de tireóides suínas e humanas. Inicialmente, analisamos as propriedades bioquímicas da NADPH-oxidase - enzima geradora de H 2 O 2 - localizada na membrana apical da célula tireóidea suína. Nossos resultados demonstram que a atividade geradora de H 2 O 2 na tireóide suína ocorre, principalmente, na fração de membrana apical tireóidea (P 3.000g). Entretanto, no P 3.000g a enzima NADPH-oxidase é apenas parcialmente cálcio-dependente, ao contrário do que acontece em frações purificadas de membrana tireóidea suína, nas quais a enzima é completamente dependente de cálcio, conforme estudos anteriores. Nossos resultados confirmam os previamente descritos para a NADPH oxidase tireóidea suína. Em tecidos tireóideos humanos, a geração de H 2 O 2 ocorreu, tanto na fração microsomal (P 100.000g) quanto na fração de membrana apical (P 3.000g). Nossos dados revelam ainda que a NADPH-oxidase humana é completamente cálcio-dependente, ativada por altas concentrações de fosfato e parece ser tão ativa na glândula humana quanto na suína. Além disto, a enzima humana é dependente de adenina-flavina-dinucleotídeo (FAD) no meio de reação, ou seja, parece ser uma flavoproteína, assim como a proteína suína.

Effect of ceramide on apoptosis and phospholipase D activity in FRTL-5 thyroid cells

Ceramide, a product of sphingomyelin hydrolysis, is now recognized as an intracellular lipid messenger, which mediates the effects of extracellular agents on cellular growth, differentiation and apoptosis. Recently, ceramide has been implicated in the regulation of phospholipase D (PLD). In this study, we examined the effects of ceramide on the activity and mRNA level of PLD during apoptotic process in FRTL-5 thyroid cells. C2-ceramide (N-acetyl sphingosine) induced apoptosis in FRTL-5 thyroid cells. Fluorescent staining showed that ceramide induced the typical features of apoptosis including condensed or fragmented nuclei. DNA fragmentation was also observed by agarose gel electrophoresis. Flow cytometric cell cycle analysis showed more clearly that ceramide induced apoptotic cell death in FRTL-5 thyroid cells. The treatment of FRTL-5 thyroid cells with thyroid-stimulating hormone (TSH) resulted in an increased PLD activity in a dose- and time-dependent manner. However, the TSH-induced increase in PLD activity was down-regulated within 2 h after ceramide treatment. Furthermore, the levels of PLD mRNA were found to be decreased throughout apoptotic process as inferred by reverse transcription-polymerase chain reaction. However, the decreases in PLD mRNA levels were not correlated with those in PLD activities after ceramide treatment. Taken together, these data suggest that ceramide inhibits the PLD activity in an early apoptotic phase and down-regulation of the levels of PLD mRNA may be implicated in apoptotic process in FRTL-5 thyroid cells.

Effects of estradiol benzoate on 5'-iodothyronine deiodinase activities in female rat anterior pituitary gland, liver and thyroid gland

There is little information on the possible effects of estrogen on the activity of 5'-deiodinase (5'-ID), an enzyme responsible for the generation of T3, the biologically active thyroid hormone. In the present study, anterior pituitary sonicates or hepatic and thyroid microsomes from ovariectomized (OVX) rats treated or not with estradiol benzoate (EB, 0.7 or 14 mug/100 g body weight, sc, for 10 days) were assayed for type I 5'-ID (5'-ID-I) and type II 5'-ID (5'-ID-II, only in pituitary) activities. The 5'-ID activity was evaluated by the release of (125)I from deiodinated (125)I rT3, using specific assay conditions for type I or type II. Serum TSH and free T3 and free T4 were measured by radioimmunoassay. OVX alone induced a reduction in pituitary 5'-ID-I (control = 723.7 + 67.9 vs OVX = 413.9 + 26.9; P<0.05), while the EB-treated OVX group showed activity similar to that of the normal group. Thyroid 5'-ID-I showed the same pattern of changes, but these changes were not statistically significant. Pituitary and hepatic 5'-ID-II did not show major alterations. The treatment with the higher EB dose (14 mug), contrary to the results obtained with the lower dose, had no effect on the reduced pituitary 5'-ID-I of OVX rats. However, it induced an imporatnt increment of 5'-ID-I in the thyroid gland (0.8 times higher than that of the normal group: control = 131.9 + 23.7 vs OVX + EB 14 mug = 248.0 + 31.2; P<0.05), which is associated with increased serum TSH (0.6-fold vs OVX, P<0.05) but normal serum free T3 and free T4. The data suggest that estrogen is a physiological stimulator of anterior pituitary 5'-ID-I and a potent stimulator of the thyroid enzyme when employed at high doses.

Thyroid peroxidase activity in human nodular goiters

1. Thyroid peroxidase (TPO, iodide-oxidation) activity was evaluated in nodular and paranodular tissue samples from 27 patients with nodular goiter (19 "cold" and 8 "hot" nodules), and compared to 11 diffuse toxic goiter and 9 normal thyroid tissue samples. 2. In terms of U/g digitonin solubilized protein, TPO activity was increased in hot nodules (p<0.05), although not as much as in diffuse toxic goiters (p<0.01> 3. The mean TPO activity of tissues paranodular to a cold nodule was not different from that of normal thyroids. 4. Both the highest and the lowest TPO activities were found in cold nodules, but their mean value did not differ from those of their paranodular tissues or normal thyroids. 5. Inter-tissue variability was significantly increased (p<0.01) in cold nodules and in tissues paranoudular to a hot nodule. 6. These data show that heterogeneity both within and among tissues contributes to the wide range of TPO activity detected in nodular goiters

Isolation and characterization of monoamine oxidase from hyperfunctioning human thyroid.

Human thyroid monoamine oxidase from hyperfunctioning thyroids was isolated and purified by separating the mitochondria by differential centrifugation followed by ultrasonication. The suspension containing the active enzyme from the DEAE-column, on passing through a Sephadex G-200 column gave three peaks corresponding to molecular weights of approximately 220,000, 23,000 and 4000 of which the component with molecular weight 4000 showed enzyme activity. The blood of the patients was analyzed for plasma MAO, RBC cholinesterase, plasma histaminase and plasma catecholamines. Histology and histochemistry of the thyroid tissues were also done. The data are examined from the point of view of the prevalent idea of the possible existence of MAO in multiple forms.