Effects of the glucocorticoid clobetasol propionate and its mixture with cortisol and different class steroids in adult female zebrafish.

Ecotoxicological effects of glucocorticoids and steroid mixtures in the environment are not sufficiently known. Here we investigate effects of 11-14 days exposure of female zebrafish to the glucocorticoid clobetasol propionate (Clo), cortisol (Cs), their mixture and mixtures with five different class steroids (Clo + triamcinolone + estradiol + androstenedione + progesterone) in liver, brain and gonads. Cs showed little activity, while Clo reduced the condition factor at 0.57 and 6.35 µg/L. Clo induced differential expression of genes in the liver at 0.07-6.35 µg/L, which were related to circadian rhythm (per1, nr1d2), glucose metabolism (g6pca, pepck1), immune system response (fkbp 5, socs3, gilz), nuclear steroid receptors (pgr and pxr), steroidogeneses and steroid metabolism (hsd11b2, cyp2k22). Clo caused strong transcriptional down-regulation of vtg. Similar upregulations occurred in the brain for pepck1, fkbp5, socs3, gilz, hsd11b2, and nr1d2a, while cyp19b was down-regulated. Effects of Clo + Cs mixtures were similar to Clo alone. Transcriptional alterations were different in mixtures of five steroids with no alteration of vtg in the liver due to counteraction of Clo and estradiol. Induction of fkbp5 (brain) and sult2st3 (liver) and downregulation of cyp19a (gonads) occurred at 1 µg/L. Histological effects of the five steroids mixture in gonads were characterized by a decrease of mature oocytes. Our data indicate that effects of steroids of different classes sum up to an overall joint effect driven by the most potent steroid Clo.

Candida albicans-Induced NETosis Is Independent of Peptidylarginine Deiminase 4.

Neutrophils are the most abundant innate immune cells and the first line of defense against many pathogenic microbes, including the human fungal pathogen Candida albicans. Among the neutrophils' arsenal of effector functions, neutrophil extracellular traps (NETs) are thought to be of particular importance for trapping and killing the large fungal filaments by means of their web-like structures that consist of chromatin fibers decorated with proteolytic enzymes and host defense proteins. Peptidylarginine deiminase 4 (PAD4)-mediated citrullination of histones in activated neutrophils correlates with chromatin decondensation and extrusion and is widely accepted to act as an integral process of NET induction (NETosis). However, the requirement of PAD4-mediated histone citrullination for NET release during C. albicans infection remains unclear. In this study, we show that although PAD4-dependent neutrophil histone citrullination is readily induced by C. albicans, PAD4 is dispensable for NETosis in response to the fungus and other common NET-inducing stimuli. Moreover, PAD4 is not required for antifungal immunity during mucosal and systemic C. albicans infection. Our results demonstrate that PAD4 is dispensable for C. albicans-induced NETosis, and they highlight the limitations of using histone citrullination as a marker for NETs and PAD4-/- mice as a model of NET-deficiency.