Деякі проблеми використання тимчасово зайнятих земель

<div><p>Glucocorticoid induced-leucine zipper (GILZ) has been shown to be induced in cells by different stimuli such as glucocorticoids, IL-10 or deprivation of IL-2. GILZ has anti-inflammatory properties and may be involved in signalling modulating apoptosis. Herein we demonstrate that wildtype <em>Yersinia enterocolitica</em> which carry the pYV plasmid upregulated GILZ mRNA levels and protein expression in epithelial cells. Infection of HeLa cells with different <em>Yersinia</em> mutant strains revealed that the protease activity of YopT, which cleaves the membrane-bound form of Rho GTPases was sufficient to induce GILZ expression. Similarly, <em>Clostridium difficile</em> toxin B, another bacterial inhibitor of Rho GTPases induced GILZ expression. YopT and toxin B both increased transcriptional activity of the GILZ promoter in HeLa cells. GILZ expression could not be linked to the inactivation of an individual Rho GTPase by these toxins. However, forced expression of RhoA and RhoB decreased basal <em>GILZ</em> promoter activity. Furthermore, MAPK activation proved necessary for profound GILZ induction by toxin B. Promoter studies and gel shift analyses defined binding of upstream stimulatory factor (USF) 1 and 2 to a canonical c-Myc binding site (E-box) in the <em>GILZ</em> promoter as a crucial step of its trans-activation. In addition we could show that USF-1 and USF-2 are essential for basal as well as toxin B induced GILZ expression. These findings define a novel way of <em>GILZ</em> promoter trans-activation mediated by bacterial toxins and differentiate it from those mediated by dexamethasone or deprivation of IL-2.</p> </div

Rifampicin-induced virulence determinants increase Candida albicans biofilm formation: XTT reduction

<p>Raw data table of absorption of reduced XTT dye after 2h incubation on SC5314 biofilm retained on a 96-well plate (background substraction has been performed).</p

Rifampicin-induced virulence determinants increase Candida albicans biofilm formation: germ tube length

<p>Experiments 1-4 of germ tube length: untreated and rifampicin treated.</p

Rifampicin-induced virulence determinants increase Candida albicans biofilm formation: hydrocarbon extraction

<p>Experiments 1-4 of hydrocarbon extraction: untreated/no extraction, untreated/xylene extraction 1 and 2, rifampicin/no extraction, and rifampicin/xylene extraction 1 and 2.</p

Reduced cell wall chitin effects late phase cytokine response to <i>C. albicans</i>.

<p>CHIT-1-activity of (A) hPMNs and (B) hMφ's incubated with live <i>C. albicans</i> yeast cells, MOI = 0.4. Values represent means ± SEM, n = 4, *p<0.05, ***p<0.001, ****p<0.0001. (C) Heat-treated yeast cells from <i>C. albicans</i> wild type and <i>chs3Δ</i> mutant were stained for total chitin content with Calcofluor White (CFW) and surface presented chitin with wheat germ agglutinin (WGA) and (D) mean fluorescence intensity (MFI) was analysed. Values are presented as mean ± SEM, n = 30, ****p<0.0001. (E) TEM analysis of heat-treated yeast cells from <i>C. albicans</i> wild type and <i>chs3Δ</i> mutant. Images shown are representative for all analysed yeast cells. (F and G) hPBMCs were incubated with heat-treated <i>C. albicans</i> wild type yeast cells or <i>C. albicans chs3Δ</i>, MOI = 0.4 in the presence or absence of the chitinase inhibitor Bisdionin C. TNF and IL-10 secretion was monitored for a period of 7 days. Values represent means ± SEM, n = 4, *p<0.05, **p<0.01, ****p<0.0001.</p

Chitin induced IL-10 secretion requires mannose receptor interaction but not uptake.

<p>(A) Chitin was incubated with liposomal transfection reagent DOTAP for 15 min at room-temperature before used to stimulate mBMMφs from wild type, TLR2, TLR9, NOD2, MR and dectin-1-deficient mice for 24 h or (A and B) mBMMφs were treated with CytD for 1 h prior chitin stimulation. Values represent mean ± SEM, n = 4, *p<0.05, **p<0.01.</p

Chitin co-localisation with NOD2 and TLR9 depends on MR.

<p>Fluorescence microscopy of mBMMφs from wild type, TLR2-, NOD2-, MR- and dectin-1-deficient mice stimulated with chitin for 1 h. Nuclei are stained with DAPI (blue) and chitin was detected using a chitin-binding reporter construct (ChBD-HuCκ, green). (A) NOD2 protein was detected with anti-mouse NOD2 antibody (red) (B) TLR9 protein was detected with anti-mouse TLR9 antibody (red) and (C) TLR2 protein was detected with anti-mouse TLR2 antibody (red). White arrows indicate co-localisation of chitin with NOD2 and/or TLR9 (yellow). Chitin did not co-localise with TLR2 in all tested cells (C) and no chitin co-localisation with NOD2 or TLR9 was detectable in MR-deficient macrophages (A and B). Images (A to C) are representative of two independent experiments, scale bars = 15 µm.</p

Chitin dampens LPS induced inflammation <i>in vivo</i>.

<p>C57BL/6 mice were injected intraperitoneal with saline, chitin, LPS or chitin and LPS in combination. Infiltrating immune cells and cytokine production were analysed after 4 h (A and D), 24 h (B and E) and 4 days (C and F). Data are presented as mean values ± SEM, n = 5 mice per group, *p<0.05, **p<0.01.</p

Chitin purity and size.

<p>(A) HPLC analysis of TFAA hydrolysed <i>C. albicans</i>-chitin (<i>left</i>) compared to commercial crab shell chitin (<i>right</i>). <i>GlcN</i> Glucosamine, <i>Glc</i> Glucose, <i>Man</i> Mannan. (B) Chitin particle size determined by flow cytometry. (C) Chitin size distribution in percentage of analysed chitin extractions, presented as mean ± SEM, n = 6.</p

Acetylation degree of purified chitin.

<p>Acetylation degree of purified chitin.</p