The influence of ghrelin on the thyroid-stimulating hormone (TSH)-induced increase in thyroglobulin (Tg) and cAMP production.

<p>The influence of ghrelin on the TSH-induced increase in Tg and cAMP production at three different concentrations of TSH (0.1 IU/L, 0.5 IU/L and 1 IU/L). The basal levels, i.e. the values in the absence of TSH, were subtracted, before the groups were compared. Grey = vehicle, pattern = ghrelin (100 nM). Means (+SEM). *P < 0.05 compared to the control (vehicle). <b>A)</b> Ghrelin inhibited the TSH-induced increase in Tg production measured by enzyme-linked immunosorbent assay (ELISA) in primary cultures of human thyroid cells for the TSH concentration of 0.1 IU/L. n = 8 (0.1 IU/L) and n = 6 (0.5 and 1 IU/L) in triplets. Two patient samples were excluded due to lack of basal TSH-induced Tg production. <b>B)</b> No influence of ghrelin on the TSH-induced increase in cAMP production at three different concentrations of TSH (0.1 IU/L, 0.5 IU/L and 1 IU/L) measured by a competitive protein binding method in primary cultures of human thyroid cells. n = 8 (0.1 IU/L and 1 IU/L) and n = 6 (0.5 IU/L) in triplets.</p

Primer sequences for RT-qPCR.

<p>Primer sequences for RT-qPCR.</p

The influence of ghrelin on the thyroid-stimulating hormone (TSH)-induced (0.1 IU/L) mRNA expression of four thyroid components.

<p>The expression of the TSH receptor (TSH-R), thyroperoxidase (TPO), thyroglobulin (Tg) and sodium iodide symporter (NIS) measured by real-time quantitative polymerase chain reaction (RT-qPCR) in a primary culture of human thyroid cells in presence and absence of ghrelin. Indicated as fold change of mRNA expression compared to basal level (dashed line). IL-6 was used as a negative control. Grey = vehicle, pattern = ghrelin (100 nM). Means (±SEM), n = 6. *P < 0.05 compared to the control (vehicle). Two patients were excluded due to unknown sample material.</p

Ghrelin receptor (GhrR) mRNA expression level.

<p>GhrR mRNA expression level in relation to the reference gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA expression level in human brain, thyroid tissue and cell cultures measured by real-time quantitative polymerase chain reaction (RT-qPCR). n = 2.</p

Influence of Phthalates on <i>in vitro</i> Innate and Adaptive Immune Responses

<div><p>Phthalates are a group of endocrine disrupting chemicals, suspected to influence the immune system. The aim of this study was to investigate the influence of phthalates on cytokine secretion from human peripheral blood mononuclear cells. <i>Escherichia coli</i> lipopolysaccharide and phytohemagglutinin-P were used for stimulation of monocytes/macrophages and T cells, respectively. Cells were exposed for 20 to 22 hours to either di-ethyl, di-n-butyl or mono-n-butyl phthalate at two different concentrations. Both diesters were metabolised to their respective monoester and influenced cytokine secretion from both monocytes/macrophages and T cells in a similar pattern: the secretion of interleukin (IL)-6, IL-10 and the chemokine CXCL8 by monocytes/macrophages was enhanced, while tumour necrosis factor (TNF)-α secretion by monocytes/macrophages was impaired, as was the secretion of IL-2 and IL-4, TNF-α and interferon-γ by T cells. The investigated phthalate monoester also influenced cytokine secretion from monocytes/macrophages similar to that of the diesters. In T cells, however, the effect of the monoester was different compared to the diesters. The influence of the phthalates on the cytokine secretion did not seem to be a result of cell death. Thus, results indicate that both human innate and adaptive immunity is influenced <i>in vitro</i> by phthalates, and that phthalates therefore may affect cell differentiation and regenerative and inflammatory processes <i>in vivo</i>.</p></div

Influence of phthalates on T-cell responses.

<p>MNC cultures were stimulated with phytohemagglutinin-P and exposed to di-ethyl phthalate (DEP), di-n-butyl phthalate (DnBP) or mono-n-butyl phthalate (MnBP), at two different concentrations, for 20–22 hours. The resulting production of IL-2, IL-4, IL-6, IL-10, TNF-α and IFN-γ are shown as ratios to the respective ethanol control. The red dashed line indicates the level of the ethanol controls (ratio = 1). * = p<0.05 compared to ethanol control, # = p<0.05 compared to low phthalate exposure (0.1 μM).</p

Influence of phthalates on the cytokine response of innate immune cells.

<p>MNC cultures were stimulated with 100 pg/ml E. coli LPS and exposed to di-ethyl phthalate (DEP), di-n-butyl phthalate (DnBP), or mono-n-butyl phthalate (MnBP), at two different concentrations, for 20–22 hours. The resulting production of IL-1β, IL-6, CXCL8, IL-10 and TNF-α are shown as ratio to the respective ethanol control. The red dashed line indicates the level of the ethanol controls (ratio = 1). * = p<0.05 compared to ethanol control, # = p<0.05 compared to low phthalate exposure (0.1 μM).</p