bluuebunny/crossref_metadata_2025_split huggingface dataset:
Antibodies to Thyroid Peroxidase Arise Spontaneously with Age in NOD.H-2h4 Mice and Appear after Thyroglobulin Antibodies
Authors: Chen Chun-Rong , Hamidi Sepehr , Braley-Mullen Helen , Nagayama Yuji , Bresee Catherine , Aliesky Holly A. , Rapoport Basil , McLachlan Sandra M. 
Published: January 2014

Abstract
Hashimoto’s thyroiditis, a common autoimmune disease, is associated with autoantibodies to thyroglobulin (Tg) and thyroid peroxidase (TPO). TPO, unlike abundant and easily purified Tg, is rarely investigated as an autoantigen in animals. We asked whether antibodies (Abs) develop to both TPO and Tg in thyroiditis in mice that is induced (C57BL/6 and DBA/1 strains) or arises spontaneously (NOD.H-2h4). Screening for TPOAbs was performed by flow cytometry using mouse TPO-expressing eukaryotic cells. Sera were also tested for binding to purified mouse Tg and human TPO. The antibody data were compared with the extent of thyroiditis. Immunization with mouse TPO adenovirus broke self-tolerance to this protein in C57BL/6 mice, but thyroiditis was minimal and TgAbs were absent. In DBA/1 mice with extensive granulomatous thyroiditis induced by Tg immunization, TPOAbs were virtually absent despite high levels of TgAbs. In contrast, antibodies to mouse TPO, with minimal cross-reactivity with human TPO, arose spontaneously in older (7–12 months) NOD.H-2h4 mice. Unexpectedly, TgAbs preceded TPOAbs, a time course paralleled in relatives of probands with juvenile Hashimoto’s thyroiditis. These findings demonstrate a novel aspect of murine and human thyroid autoimmunity, namely breaking B cell self-tolerance occurs first for Tg and subsequently for TPO.
Wnt/β-Catenin Pathway Activation in Adrenocortical Adenomas Is Frequently due to Somatic CTNNB1-Activating Mutations, Which Are Associated with Larger and Nonsecreting Tumors: A Study in Cortisol-Secreting and -Nonsecreting Tumors
Authors: Bonnet Stéphane , Gaujoux Sébastien , Launay Pierre , Baudry Camille , Chokri Ilham , Ragazzon Bruno , Libé Rossella , René-Corail Fernande , Audebourg Anne , Vacher-Lavenu Marie-Cécile , Groussin Lionel , Bertagna Xavier , Dousset Bertrand , Bertherat Jérôme , Tissier Frédérique 
Published: January 2014

Background
Abnormal β-catenin immunohistochemistry and mutations of the β-catenin gene (CTNNB1) have been reported in adrenocortical adenomas (ACAs), but the frequencies of these defects and the phenotype of such tumors have not been clearly determined.

Objective
The objective of the study was to describe the Wnt/β-catenin pathway alterations in 100 ACAs and their association with clinicopathological characteristics.

Patients and methods
One hundred consecutive ACAs (excluding Conn’s adenomas) were studied clinically by β-catenin immunohistochemistry and direct sequencing of CTNNB1.

Results
Thirty-five ACAs were nonsecreting adenomas (NSAs), 19 were subclinical cortisol secreting adenomas (SCSAs), and 46 were cortisol secreting adenomas (CSAs). Fifty-one tumors had abnormal cytoplasmic and/or nuclear β-catenin immunohistochemical staining, indicating Wnt/β-catenin pathway alteration. Thirty-six tumors showed CTNNB1 mutations, which all showed abnormal immunohistochemical β-catenin accumulation. Among the 64 nonmutated tumors, only 15 (23%) showed cytoplasmic and/or nuclear β-catenin staining (P &lt; 0.0001). Tumors with CTNNB1 mutations were predominantly nonsecreting (61% NSAs, 22% SCSAs, 16% CSAs) whereas nonmutated tumors were predominantly secreting (20% NSAs, 17% SCSAs, 62% CSAs) (P &lt; 0.0001). Mean tumor size and weight were, respectively, 4.2 cm (±1.2) and 28.4 g (±21.1) for tumors with CTNNB1 mutations vs. 3.4 cm (±0.9) and 18.2 g (±8.1) for nonmutated tumors (P &lt; 0.01).

Conclusions
Abnormal cytoplasmic and/or nuclear β-catenin immunohistochemical staining occurs in about half of ACAs. This suggests the activation of the Wnt/β-catenin pathway, which could be explained by activating mutations of CTNNB1 in 70% of the cases. CTNNB1 mutations are mainly observed in larger and nonsecreting ACAs, suggesting that the Wnt/β-catenin pathway activation is associated with the development of less differentiated ACAs.
Is the Vitamin D Receptor Found in Muscle?
Authors: Wang Yongji , DeLuca Hector F. 
Published: January 2014

Abstract
The active form of vitamin D, 1α,25-dihydroxyvitamin D3, is critical for the regulation of serum calcium and phosphorus levels that in turn support bone mineralization and neuromuscular activity. It is well known that vitamin D deficiency causes rachitic/osteomalacic myopathy and cardiac disorder and the provision of vitamin D can reverse the symptoms. However, the underlying mechanisms remain unclear. The question of whether the vitamin D receptor is found in muscle has been debated but not settled. We recently studied all available antibodies against the vitamin D receptor and found that most antibodies used detect proteins other than the vitamin D receptor, and therefore, the utility of these antibodies may generate the false-positive results. Using antibodies that do not detect proteins in tissues from vitamin D receptor null mice, we have developed a specific and sensitive immunohistochemical assay. The results from this investigation show that the vitamin D receptor is undetectable in skeletal, cardiac, and smooth muscle, suggesting that the function of vitamin D on muscle is either of an indirect nature or does not involve the known receptor.
Inactivation of Melatonin Receptors by Protein Kinase C in Human Prostate Epithelial Cells
Authors: Gilad Eli , Matzkin Haim , Zisapel Nava 
Published: January 2014

AbstractThe pineal hormone melatonin regulates seasonal reproduction and pubertal development in mammals. We recently found melatonin receptors in the human benign prostate tissue, primarily associated with the microsome-enriched fraction of the epithelial cells. In cultured benign prostate epithelial cells, melatonin, at physiological concentrations, suppressed [3H]thymidine incorporation and cGMP levels. The effects of melatonin were transient, suggesting inactivation of the receptors. In the present study, the possibility of inactivation of the prostate melatonin receptors by protein kinase C (PKC) was explored.Treatment of the microsome-enriched fraction with crude rat brain PKC in the presence of phorbol 12-myristate 13-acetate (TPA) or CaCl2 abolished the specific [125I]melatonin binding. This effect was prevented by the PKC inhibitor bisindolylmaleimide (GF-109203). [125I]Melatonin binding could be reinstated by iodoacetamide treatment.In benign prostate epithelial cells in culture, TPA pretreatment markedly reduced the apparent affinity of [125I]melatonin binding. In addition, TPA ablated the cells responses to melatonin, namely the suppression of [3H]thymidine incorporation and cGMP levels. Pretreatment with GF-109203 prevented the TPA effects on[ 125I]melatonin binding and responses. In addition, GF-109203 slowed down the inactivation of the melatonin-mediated inhibition of [3H]thymidine incorporation.Taken together, these data show that melatonin receptors are desensitized by PKC and imply that the transient response to melatonin may be the outcome of a direct or indirect melatonin-mediated activation of endogenous PKC.
Effect of Endothelin-1 on Corticosteroid Secretion by the Frog Adrenal Gland Is Mediated by an EndothelinA Receptor*
Authors: Cartier Franck , Remy-Jouet Isabelle , Fournier Alain , Vaudry Hubert , Delarue Catherine 
Published: January 2014

AbstractWe have previously reported that endothelin-1 (ET-1) stimulates the in vitro secretion of corticosterone and aldosterone from the adrenal gland of the frog Rana ridibunda. The aim of the present study was to investigate the pharmacological profile of the endothelin receptor subtype involved in the corticotropic effect of ET-1. The mixed ETA/ETB receptor antagonist Ro 47–0203 (10−5m) totally blocked the stimulatory effect of ET-1 (5 × 10−9m) on corticosterone and aldosterone secretion. The action of ET-1 was also inhibited by the selective ETA receptor antagonist BQ-485 (10−7m). In contrast, the selective ETB receptor antagonist IRL 1038 (10−6m) did not affect the response of the frog adrenal gland to ET-1. In addition, the selective ETB receptor agonist IRL 1620 (10−6m) did not mimic the stimulatory effect of ET-1. The high affinity ETC receptor agonist endothelin-3 (ET-3) stimulated corticosteroid secretion, but was 400 times less potent than ET-1. Moreover, the action of ET-3 was also blocked by BQ-485 (10−7m). These data indicate that the stimulatory effects of ET-1 and ET-3 on corticosteroid secretion by the frog adrenal gland are mediated by an ETA receptor subtype.
Maternal Adrenalectomy Eliminates a Surge of Plasma Dehydroepiandrosterone in the Mother and Attenuates the Prenatal Testosterone Surge in the Male Fetus*
Authors: Sinha Partha , Halasz Ildiko , Choi Jennifer F. , McGivern Robert F. , Redei Eva 
Published: January 2014

Abstract
Previous work has established a number of sex-related deficits in immune function, behavior, and endocrine responses to stress in the offspring of dams exposed to ethanol. To examine the potential role of maternal glucocorticoids as a mediator of these sexually dimorphic effects in the fetus, we examined the influence of prenatal alcohol exposure in the presence or absence of maternal glucocorticoids on fetal plasma corticosterone (CORT) production. An additional question to be addressed by these studies was whether maternal adrenalectomy could eliminate the known inhibition by ethanol of the prenatal surge of plasma testosterone in male fetuses.
Pregnant dams were adrenalectomized (ADX) or sham-adrenalectomized on gestational day (G) 7 and placed on a liquid diet containing 35% ethanol-derived calories or pair-fed an isocaloric control diet throughout the experiment. On G18, G19, and G21, plasma levels of CORT, testosterone, and dehydroepiandrosterone (DHEA) were measured in male and female fetuses and their mothers. Ethanol administration consistently increased maternal plasma CORT levels but did not significantly alter CORT levels in the fetus. Maternal ADX resulted in compensatory increases in fetal CORT levels that were lower in fetuses of ADX dams on alcohol, suggesting a direct effect of ethanol on fetal pituitary-adrenal activity. There were no significant sex differences in fetal plasma CORT levels in response to any of these manipulations.
A novel surge of maternal plasma DHEA was found on G19 that was absent in plasma from ADX dams. In spite of the absence of a surge on G19, plasma DHEA levels of ADX dams rose from very low levels at G18 to levels on G21 that were significantly higher than in Sham dams. A normal testosterone surge was observed in male fetuses on G18 and G19 from sham-adrenalectomized dams administered the pair-fed diet. However, this surge was greatly attenuated in males administered ethanol and also in male fetuses from ADX dams. These results reveal a direct inhibitory influence of ethanol on fetal CORT secretion as well as on the prenatal testosterone surge in males. Furthermore, these studies demonstrate the presence of a surge of DHEA in the pregnant rat. Overall, these data suggest that there is a critical adrenal factor in the rat that regulates the maternal surge of DHEA on G19 and the prenatal testosterone surge of male fetuses on G18–19.
Effects of Antagonists of Growth Hormone-Releasing Hormone (GHRH) on GH and Insulin-Like Growth Factor I Levels in Transgenic Mice Overexpressing the Human GHRH Gene, an Animal Model of Acromegaly*
Authors: Kovacs Magdolna , Kineman Rhonda D. , Schally Andrew V. , Zarandi Marta , Groot Kate , Frohman Lawrence A. 
Published: January 2014

Abstract
Transgenic mice overexpressing the human GH-releasing hormone (hGHRH) gene, an animal model of acromegaly, were used to investigate the effects of potent GHRH antagonists MZ-4–71 and MZ-5–156 on the excessive GH and insulin-like growth factor I (IGF-I) secretion caused by overproduction of hGHRH. Because metallothionein (MT)-GHRH mice express the hGHRH transgene in various tissues, including the pituitary and hypothalamus, initial experiments focused on the effectiveness of the GHRH antagonists in blocking basal and stimulated GH secretion from pituitary cells in vitro. Both MZ-4–71 and MZ-5–156 suppressed basal release of GH from superfused MT-GHRH pituitary cells, apparently by blocking the action of endogenously produced hGHRH. In addition, these antagonists effectively eliminated the response to stimulatory action of exogenous hGHRH(1–29)NH2 (30 and 100 nm). To ascertain whether MZ-4–71 and MZ-5–156 could antagonize the effect of hGHRH hyperstimulation in vivo, each antagonist was administered to MT-GHRH transgenic mice in a single iv dose of 10–200 μg. Both compounds decreased serum GH levels in transgenic mice by 39–72% at 1 h after injection. The inhibitory effect of 50 μg MZ-5–156 was maintained for 5 h. Twice daily ip administration of 100 μg MZ-5–156 for 3 days suppressed the highly elevated serum GH and IGF-I concentrations in transgenic mice by 56.8% and 39.0%, respectively. This treatment also reduced IGF-I messenger RNA levels in the liver by 21.8% but did not affect the level of GH messenger RNA in the pituitary. Our results demonstrate that GHRH antagonists MZ-4–71 and MZ-5–156 can inhibit elevated GH levels caused by overproduction of hGHRH. The suppression of circulating GH concentrations induced by the antagonists seems to be physiologically relevant, because both IGF-I secretion and synthesis also were reduced. Our findings, showing the suppression of GH and IGF-I secretion with GHRH antagonists, suggest that this class of analogs could be used for the diagnosis and therapy of disorders characterized by excessive GHRH secretion.
Thyroid Hormones Inhibit Type 2 Iodothyronine Deiodinase in the Rat Cerebral Cortex by Both Pre- and Posttranslational Mechanisms*
Authors: Burmeister Lynn A. , Pachucki John , St. Germain Donald L. 
Published: January 2014

AbstractThe type 2 5′-deiodinase (D2) appears to play an important role in maintaining the intracerebral T3 content relatively constant during changes in thyroidal state. Previous studies have demonstrated that the regulation of this enzyme by thyroid hormone and its analogs occurs at a posttranslational level. The availability of the rat D2 complementary DNA now allows an assessment of whether pretranslational regulation of this enzyme also occurs in the cerebral cortex. In rats rendered hypothyroid by the addition of methimazole to the drinking water, D2 messenger RNA (mRNA) is increased 70% (P = 0.03). Treatment with l-T3 (50 μg/100 g BW) for 4 days results in an 80% decrease in D2 mRNA compared with that in euthyroid controls (P &lt; 0.001). Administration of lower doses of l-T3 (0.25–3 μg/100 g BW·day) is associated with a dose-dependent decrease in cortical D2 mRNA, but little or no change in D2 activity. The decrease in D2 mRNA in response to T3 treatment can be demonstrated within 4 h. Treatment of hypothyroid rats for 2 weeks with graded doses of l-T4 (0.1–1.5 μg/100 g BW·day) results in a significant decrease in cortical D2 activity, but not mRNA. The association between D2 activity and D2 mRNA in euthyroid, hypothyroid, and hormone-treated rats across a full range of thyroidal states suggests that l-T4 treatment is associated with greater changes in cortical D2 activity (via posttranslational effects) than mRNA, whereas l-T3 treatment has a greater effect on decreasing D2 mRNA (i.e. pretranslational effects). In conclusion, these studies demonstrate both pre- and posttranslational regulation of cortical D2 expression. The relative contribution of each mechanism depends on the ambient thyroid hormone concentration.
Two Frog Melanotrope Cell Subpopulations Exhibiting Distinct Biochemical and Physiological Patterns in Basal Conditions and under Thyrotropin-Releasing Hormone Stimulation*
Authors: Gonzalez de Aguilar Jose Luis , Malagon Maria M. , Vazquez-Martinez Rafael M. , Lihrmann Isabelle , Tonon Marie-Christine , Vaudry Hubert , Gracia-Navarro Francisco 
Published: January 2014

AbstractCell heterogeneity designates the phenomenon by which a particular cell type is composed of morphologically and physiologically distinct cell subpopulations. We have previously isolated two subsets of melanotrope cells in the intermediate lobe of the frog pituitary by means of a separation procedure based on a Percoll density gradient. High density (HD) melanotrope cells were found to exhibit a more granulated cytoplasm and a lower secretory rate than low density (LD) cells. In the present study, we have investigated the biochemical and functional characteristics of each melanotrope cell subpopulation by using various approaches, including chromatographic analysis for the measurement of the proportion of acetylated αMSH, microfluorimetric measurement of the cytosolic free calcium concentration ([Ca2+]i), and in situ hybridization for quantification of POMC messenger RNA (mRNA). Under basal conditions, LD melanotrope cells showed higher secretory activity, acetylation rate, [Ca2+]i, and POMC mRNA content compared to HD cells. Incubation of the cells with 100 nm TRH for 2 h induced a more pronounced activation ofα MSH secretion, [Ca2+]i mobilization, and POMC mRNA accumulation in LD than in HD melanotrope cells. Conversely, TRH increased the rate of acetylation of αMSH in HD cells, but did not affect acetylation in LD cells. Taken together, these results demonstrate that the frog intermediate lobe is composed of two subsets of endocrine cells with distinct biochemical and functional characteristics. The coexistence of two cell subpopulations in the frog pars intermedia is consistent with the idea of a cell secretory cycle, in which each melanotrope subset represents a specific state of cellular activity.
Reconstitution of Calcium-Regulated Parathyroid Hormone Secretion from Streptolysin-O-Permeabilized Parathyroid Cells by Guanosine 5′-O-(Thio)Triphosphate*
Authors: Matovcik Lisa M. , Rhee Steven S. , Schaefer Jean F. , Kinder Barbara K. 
Published: January 2014

AbstractIntracellular Ca2+ levels determine the amount of PTH secretion from parathyroid cells. Dissociated calf parathyroid cells were permeabilized with streptolysin-O (SLO) to provide an in vitro model system to examine Ca2+-dependent regulation of hormone secretion. PTH release from these cells was energy dependent and increased by cytosolic cofactors. Guanosine 5′-O-(thio)triphosphate (GTPγS) increased PTH secretion from SLO-permeabilized cells in a dose-dependent manner from 0.1–100 μm. In the absence of GTPγS there was no relationship between the ambient Ca2+ concentration and the rate of PTH secretion. However, in the presence of GTPγS, intracellular Ca2+ inhibited PTH secretion with an EC50 of approximately 0.1 μm, corresponding to physiological intracellular Ca2+ levels. Thus, the addition of GTPγS to SLO-permeabilized parathyroid cells reconstituted the inverse relationship between extracellular Ca2+ and PTH secretion that is observed in vivo and in intact cells. The data indicate that this effect is mediated at least in part by heterotrimeric guanosine triphosphatases. In addition, calcium/calmodulin-dependent protein kinase II appears to mediate low Ca2+-dependent PTH secretion from these cells.
Basal and Growth Hormone-Induced Hepatic Messenger Ribonucleic Acid Expression of Insulin-Like Growth Factor-I (IGF-I) and IGF-Binding Protein-3 Is Independent of Hyperinsulinemia and Increased Energy Status in the Genetically Obese Zucker Rat*
Authors: Melián E. , Velasco B. , Barrios R. , Sanchez-Franco F. 
Published: January 2014

Abstract
Genetically obese Zucker rats, like obese humans, have normal or elevated circulating insulin-like growth factor-I (IGF-I) levels in the presence of low GH secretion. Hyperinsulinemia, increased energy status, or other nutritional factors associated with obesity could be responsible for these findings directly by increasing hepatic IGF-I production at the transcriptional or posttranscriptional level. Alternatively, circulating IGF-I could be modulated indirectly by affecting its binding proteins. To further elucidate this point, we quantitated hepatic IGF-I, IGF binding protein-3 (IGFBP-3), and GH receptor messenger RNAs (mRNAs) expression in obese Zucker rats under different serum GH and insulin conditions using lean rats as controls. Eleven-week-old male rats were studied basally (intact) or after hypophysectomy (hx) at 9 weeks. In each condition, animals were killed before or 6 h after one dose of recombinant human GH (1.5 μg/g body weight ip). At this time, in addition to the mRNA expression of the above-mentioned genes, body weight, glycemia, insulinemia, serum GH (rat and human), and serum IGF-I levels were determined. Obese Zucker rats were significantly heavier than controls in all the conditions studied and did not show differences in glycemia. Severely hyperinsulinemic intact obese rats (146.9 ± 14 vs. 46.3 ± 3 μU/ml, P &lt; 0.001) showed compared with intact lean rats significantly lower serum GH (2.39 ± 0.9 vs. 4.98 ± 0.68 ng/ml, P &lt; 0.01), decreased hepatic IGF-I mRNA and IGFBP-3 mRNA accumulation (IGF-Ia: 79 ± 5.9% vs. 100 ± 0.9%, P &lt; 0.05; IGF-Ib: 67 ± 5.5% vs. 100.1 ± 1.9%,P &lt; 0.001; IGFBP-3: 54.7 ± 2.75% vs. 100.5 ± 1.55%, P &lt; 0.001), and similar circulating IGF-I levels (1439 ± 182 vs. 1516 ± 121 ng/ml). Under comparable serum GH levels in GH-treated intact, hx, and GH-treated hx animals, hyperinsulinemia and/or increased body weight present in obese rats were not associated with increased hepatic IGF-I and IGFBP-3 mRNA amount. No differences in GH receptor/GH-binding protein mRNAs were found in any experimental condition. These results suggest that in vivo the imbalance of the serum GH/IGF-I axis present in obesity is primarily due to events distal to the hepatic IGF-I and IGFBP-3 mRNAs expression, which is tightly correlated to GH levels.