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 Cytokine Production in Monocytes and Macrophages: A Systematic Review of Experimental Trials

<div><p>Background</p><p>Phthalates are a group of endocrine disrupting chemicals suspected to influence the immune system. The aim of this systematic review is to summarise the present knowledge on the influence of phthalates on monocyte and macrophage production and secretion of cytokines, an influence which could affect both pro- and anti-inflammatory abilities of these cells.</p><p>Strategy and Results</p><p>A systematic search was performed in Medline, Embase and Toxline in June 2013, last updated 3rd of August 2014. Criteria used to select studies were described and published beforehand online on Prospero (<a href="http://www.crd.york.ac.uk/NIHR_PROSPERO" target="_blank">http://www.crd.york.ac.uk/NIHR_PROSPERO</a>, registration number CRD42013004236). <i>In vivo</i>, <i>ex vivo</i> and <i>in vitro</i> studies investigating the influence of phthalates on cytokine mRNA expression and cytokine secretion in animals and humans were included. A total of 11 reports, containing 12 studies, were found eligible for inclusion. In these, a total of four different phthalate diesters, six primary metabolites (phthalate monoesters) and seven different cytokines were investigated. Though all studies varied greatly in study design and species sources, four out of five studies that investigated di-2-ethylhexyl phthalate found an increased tumour necrosis factor-α secretion/production from monocytes or macrophages. A summary of cytokine measurements was not possible since few studies were comparable in study design and due to insufficient reporting of raw data for most of the included studies.</p><p>Conclusion</p><p>Results from this review have suggested that at least one phthalate (di-2-ethylhexyl phthalate) has the ability to enhance tumour necrosis factor-α production/secretion from monocytes/macrophages <i>in vitro</i>, but also observed <i>ex vivo</i>. Influence of other phthalates on other cytokines has only been investigated in few studies. Thus, <i>in vitro</i> studies on primary human monocytes/macrophages as well as more <i>in vivo</i> studies are needed to confirm or dispute these findings.</p></div

Primary and secondary outcomes from individual studies.

<p>Footnote: BBzP: Butylbenzyl phthalate, DEHP: Di-2-ethylhexyl phthalate, DiNP: Di-iso-nonyl phthalate, DnBP: Di-n-butyl phthalate, IL: Interleukin, LDH: lactate dehydrogenase, MEHP: Mono-(2-ethylhexyl) phthalate, ND: not done, PI: Propidium Iodide, RAW 264 cell line: mouse leukemic monocyte-macrophage cell line, SD: standard deviation, SEM: standard error of the mean, THP-1 cell line: acute monocytic cell line, TNF: Tumour necrosis factor.</p><p>Primary and secondary outcomes from individual studies.</p

Study characteristics.

<p>Footnote:</p><p>*: macrophage/monocyte content not confirmed, but cells stimulated with LPS.</p><p>†: macrophage content confirmed by flow cytometry.</p><p>BBzP: Butylbenzyl phthalate, DEHP: Di-2-ethylhexyl phthalate, DiNP: Di-iso-nonyl phthalate, DnBP: Di-n-butyl phthalate, IL: interleukin, MBzP: Mono-benzyl phthalate, MEHP: Mono-(2-ethylhexyl) phthalate, MiDP: Mono-iso-decyl phthalate, MiNP: Mono-iso-nonyl phthalate, MnBP: Mono-n-butyl phthalate, MOP: Mono-n-octyl phthalate, NRCT: Non randomised controlled trial, PMA: Phorbol 12- myristate 13-acetate, RAW 264 cell line: mouse leukemic monocyte-macrophage cell line, RCT: Randomised controlled trial, THP-1 cell line: acute monocytic cell line, TNF: tumour necrosis factor.</p><p>Study characteristics.</p

Echocardiographic and clinical findings in patients with Fabry disease during long-term enzyme replacement therapy: a nationwide Danish cohort study

<p><i>Objectives</i>: In patients with Fabry disease (FD), left ventricular hypertrophy and arrhythmias are frequently observed and cardiac involvement is the leading cause of death. Long-term efficacy of enzyme replacement therapy (ERT) on cardiac involvement is unclear. We assessed and compared long-term progression of cardiac involvement according to ERT and non-ERT. <i>Methods</i>: We retrospectively assessed and compared long-term progression of cardiac involvement in adult patients with FD in the nationwide Danish cohort. We followed clinical signs, symptoms and findings by echocardiography, electrocardiography and Holter-monitoring. <i>Results</i>: We included 66 patients; 47 patients (27 women) received ERT (ERT group) and 19 patients (15 women) did not (non-ERT group). The groups were followed for a median of 8 [0–12] years and 6 [0–13] years, respectively. Comparison between ERT and non-ERT receiving patients by left ventricular mass (echocardiographic assessment) and Sokolow-Lyon voltage- and Cornell product criteria (electrocardiographic assessment) revealed no significant differences. In the ERT group, we observed no change in left ventricular mass but a decrease in Sokolow-Lyon voltage- and Cornell product criteria from baseline to follow-up; 30 mm [15–53] vs. 25 mm [3–44], <i>p</i> < 0.005 and 1710 mm·ms [480–3740] vs. 1520 mm·ms [550–5740], <i>p</i> < .05, respectively. There were no changes within the non-ERT group. During follow-up, cardiac symptoms and use of cardiovascular procedures and -medication increased significantly in the ERT group, whereas no differences were observed within the non-ERT group. <i>Discussion</i>: We raise concerns regarding the efficacy and benefit of ERT on cardiac involvement in Fabry disease and stress the need for further research.</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 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