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Background:Both expression of the thyrotropin receptor (TSHR) and the production of hyaluronan (HA) by orbital fibroblasts (OF) have been proposed to be implicated in the pathogenesis of Graves' ophthalmopathy (GO). HA is synthesized by three types of HA synthase. We hypothesized that TSHR activation by recombinant human TSH (rhTSH) and TSHR-stimulating Graves' disease immunoglobulins (GD-IgGs) via induced cyclic adenosine monophosphate (cAMP) signaling increases HA synthesis in differentiated OF from GO patients.Methods:Cultured human OF, obtained during decompression surgery from 17 patients with severe GO, were stimulated in vitroto differentiate into adipocytes. Differentiation was evaluated by phase-contrast microscopy. The differentiated OF were stimulated by rhTSH or by TSHR-stimulating GD-IgG. We measured cAMP using a biochemical assay, HA synthase mRNA expression by quantitative polymerase chain reaction, and HA in the supernatant by enzyme-linked immunosorbent assay.Results:All differentiated OF cultures expressed higher levels of TSHRmRNA than nondifferentiated OF cultures. Stimulation by rhTSH induced a marked cAMP response in 11 of 12 differentiated OF cultures, but no measurable HA response in all but one differentiated OF cultures. By contrast, stimulation by GD-IgG induced a moderate cAMP response in a number of differentiated OF cultures, but a marked HA response in the majority of differentiated OF cultures.Conclusion:Stimulation of differentiated OF by GD-IgG, but not by rhTSH, induces HA synthesis in the majority of patients, suggesting that in most patients TSHR-mediated cAMP signaling does not play a pivotal role in GD-IgG-induced HA synthesis in differentiated OF cultures. [ABSTRACT FROM AUTHOR]

Background:Orbital fibroblasts are involved in the pathogenesis of Graves' ophthalmopathy (GO) by producing hyaluronan (HA), synthesized by three types of hyaluronan synthases (HAS1, HAS2, and HAS3). Thyrotropin receptors (TSHR) expressed in orbital fibroblasts activate the cyclic adenosine monophosphate (cAMP) pathway. Only sparse data are available at present supporting a role for TSHR activation in the regulation of HA in GO orbital fibroblasts. We hypothesize that TSHR activation, via cAMP signaling, results in induction of HAS1–3mRNA expression and HA production by nondifferentiated GO orbital fibroblasts.Methods:Cultured nondifferentiated orbital fibroblasts obtained during orbital decompression surgery from 15 GO patients were stimulated with recombinant human TSH (rhTSH), TSHR-stimulating Graves' disease immunoglobulin G (GD-IgG) or forskolin (FSK), or interleukin-1β (IL-1β).Results:FSK significantly stimulated cAMP production, HAS1and HAS3mRNA expression, and HA secretion in orbital fibroblasts. IL-1β slightly induced cAMP production, but induced HASmRNA expression of all three isoforms and HA secretion. In contrast, the effects of rhTSH and GD-IgG on cAMP were modest and absent, respectively, and on HASmRNA and HA synthesis were completely absent.Conclusions:The strong increase in cAMP synthesis by FSK in nondifferentiated GO orbital fibroblasts results in increased HA synthesis, but TSHR activation by rhTSH or GD-IgG does not result in altered HA synthesis. Our results do not support a predominant role for GD-IgGs in the accumulation of orbital glycosaminoglycans; cytokines like IL-1β seem largely responsible for excessive glycosaminoglycan production by nondifferentiated orbital fibroblasts in early immunopathogenesis of GO. [ABSTRACT FROM AUTHOR]

Background:We have shown substantial expression of type 3 deiodinase (D3, a major enzyme involved in the inactivation of thyroid hormone) in infiltrating leukocytes in several models of inflammation. Recently, thyroid hormone has been shown to improve remyelination in experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis. As induction of D3 may play an important role in decreasing local bioavailability of thyroid hormone at inflammation sites, we hypothesized that D3 is induced in spinal cord inflammatory lesions in EAE.Methods:The aim of the study was to evaluate D3 expression in spinal cord inflammatory lesions of EAE Dark Agouti rats and to investigate D3 induction in activated monocytes.Results:Here, we show marked expression of D3 by granulocytes and macrophages in spinal cord inflammatory lesions of EAE rats. We further confirm induction of D3 expression in vitroin monocytes that were activated toward proinflammatory or immunomodulatory phenotypes.Conclusions:We observed increased D3 expression both in spinal cord inflammatory lesions during EAE and in activated monocytes. Although increased D3 expression theoretically results in decreased triiodothyronine availability, it is unknown at present whether reduced local triiodothyronine concentrations are involved in impaired remyelination as observed during EAE. [ABSTRACT FROM AUTHOR]

The transition from the fed to the fasted resting state is characterized by, among other things, changes in lipid metabolism and peripheral insulin resistance. Acylcarnitines have been suggested to play a role in insulin resistance, as well as other long-chain fatty acid metabolites. Plasma levels of long-chain acylcarnitines increase during fasting, but this is unknown for muscle long-chain acylcarnitines. In the present study we investigated whether muscle long-chain acylcarnitines increase during fasting and we investigated their relationship with glucose/fat oxidation and insulin sensitivity in lean healthy humans. After 14 h and 62 h of fasting, glucose fluxes, substrate oxidation, and plasma and muscle acylcarnitines were measured before and during a hyperinsulinaemic–euglycaemic clamp. Hyperinsulinaemia decreased long-chain muscle acylcarnitines after 14 h of fasting, but not after 62 h of fasting. In both the basal state and during the clamp, glucose oxidation was lower and fatty acid oxidation was higher after 62 h compared with 14 h of fasting. Absolute changes in glucose and fatty acid oxidation in the basal compared with hyperinsulinaemic state were not different. Muscle long-chain acylcarnitines did not correlate with glucose oxidation, fatty acid oxidation or insulin-mediated peripheral glucose uptake. After 62 h of fasting, the suppression of muscle long-chain acylcarnitines by insulin was attenuated compared with 14 h of fasting. Muscle long-chain acylcarnitines do not unconditionally reflect fatty acid oxidation. The higher fatty acid oxidation during hyperinsulinaemia after 62 h compared with 14 h of fasting, although the absolute decrease in fatty acid oxidation was not different, suggests a different set point. [ABSTRACT FROM AUTHOR]

Background:Macrophages and polymorphonuclear cells (PMNs) play an important role in the first line of defense against bacteria by infiltrating the infected organ in order to clear the harmful pathogen. Our earlier studies showed that granulocytes express type 3 deiodinase (D3) when activated during a turpentine-induced abscess. We hypothesized that D3 expression by granulocytes may also occur during bacterial infection.Methods:In order to test this hypothesis, we used the following experimental infection models: peritonitis induced by Escherichia coliand acute pneumonia induced by Streptococcus pneumoniae.Results:E. coli–induced peritonitis was characterized by infiltration in the liver by inflammatory cells with abundant immunocytochemical D3 expression while no staining was present in hepatocytes of infected or control mice. Acute pneumonia induced by S. pneumoniaeresulted in inflamed lungs characterized by numerous infiltrating granulocytes expressing D3 while no D3 staining was present in lung sections without an infiltrate. Serum thyroid hormones were negatively correlated to bacterial outgrowth in both lung and spleen, and thus to the severity of illness.Conclusion:Infiltrating granulocytes during acute bacterial infection express D3. Our work supports the hypothesis that D3 plays an important role during chemical and bacterial inflammation. Whether the resulting decreased local bioavailability of thyroid hormones or rather the increased local availability of iodide is an important element of the innate immune response remains to be studied. [ABSTRACT FROM AUTHOR]

Fasting induces profound changes in the hypothalamuspituitarythyroid (HPT) axis. The alterations observed in humans and rodents are similar in many ways, although they may be more pronounced and more acute in rodents. The molecular mechanisms underlying the resetting of HPT axis regulation in the framework of caloric deprivation are still incompletely understood. Fascinating studies in rats and mice have shown a dramatic downregulation of thyrotropin-releasing hormone (TRH) gene expression in hypophysiotropic paraventricular nucleus (PVN) neurons during fasting. Direct and indirect effects of decreased serum leptin, as well as effects of increased local triiodothyronine (T3) concentrations, in the hypothalamus during food deprivation contribute to the decreased activity of TRH neurons in the PVN. However, the relative contributions of these complex determinants remain to be defined in more detail. Pituitary thyroid-stimulating hormone (TSH) mRNA expression decreases during fasting, and this may be relatively independent of leptin and/or TRH, since leptin administration in this setting does not fully restore pituitary TSH expression, while it does restore TRH expression in the PVN. There may be a role for pituitary peptides, such as neuromedin B, in altered TSH gene expression during fasting. The observed decrease in serum thyroid hormone concentrations results to some extent from diminished thyroidal secretion of thyroid hormones, especially in rodents. Decreased thyroxine (T4) and T3 contribute to the downregulation of T3-responsive genes such as liver D1. The overall result of these complex HPT axis changes in various tissues during fasting is downregulation of the HPT axis, which is assumed to represent an energy-saving mechanism, instrumental in times of food shortage. [ABSTRACT FROM AUTHOR]

Objective:The THRA gene encodes two isoforms of the thyroid hormone receptor (TR), TRα1 and TRα2. The ratio of these splice variants could have a marked influence on T3–regulated gene expression, especially during illness. Design:We studied the expression of the isoforms TRβ1, TRα1, and TRα2 and 5′-deiodinase in postmortem liver biopsies of 58 patients who were critically ill and died in the intensive care unit (ICU). All mRNA levels were determined using real-time PCR. Main outcome:All ratios of the biopsies were higher than those found in three normal liver biopsies due to an increased TRα1 level. The TRα1TRα2 ratio increased with age and severity of illness following the equation: TRα1TRα2 ratio − 1.854 (0.0323 × age) (0.0431 × Therapeutic Intervention Scoring System score) indicating that 28% of the changed TRα1TRα2 ratio can be predicted by these clinical variables. There was no effect of randomization to intensive insulin therapy or glucocorticoid or thyroid hormone treatment on the TRα1TRα2 ratio or TRβ1. Furthermore, no relation was seen between the expression levels of the 5′-deiodinase mRNA and TR isoforms or the triiodothyronine (T3) levels. Conclusion:It appears that in critically ill patients the ratio of TRα1TRα2 expression increases with age and severity of illness, possibly indicating a mechanism to enhance sensitivity to T3in the oldest and sickest patients. [ABSTRACT FROM AUTHOR]