Involvement of ERK1/2 MAPK in IL-6 and IL-8 release.

<p>(A, B) Cells were treated with UDP alone or co-incubated with ERK1/2 MAPK inhibitor, PD 98059 (50 µM), for 6 h. IL-6 (A) and IL-8 (B) levels were measured using an ELISA. Levels of IL-6 and IL-8 were corrected with vehicle control alone and normalized to UDP-stimulated cytokine release. Each column represents the mean ± S.E. (<i>n</i> = 4–6). Statistically significant inhibitory effects compared with UDP-treated controls are marked with an asterisk (*<i>p</i><0.05, Student’s <i>t</i>-test). (C) Effect of PD 98059 (50 µM) alone on basal IL-6 (16HBE14o- cells) or IL-8 (16HBE14o- cells and primary HBE cells) release. Each column represents the mean ± S.E. (<i>n</i> = 4–6). (D) The 16HBE14o- cells were stimulated with 100 µM UDP for the indicated times. Protein matched lysates were analyzed by western blotting with antibodies against phospho-ERK1/2 and ERK1/2 MAPK. The graph represents fold change in phosphorylation signal over total MAPK, normalized to GAPDH. The phosphoMAPK∶total MAPK ratio of the vehicle control was arbitrarily set to 1. Data shown are representative of six independent experiments (*<i>p</i><0.05 compared with vehicle control, Student’s <i>t</i>-test). (E) Similar western blot experiments were performed to show the inhibitory effect of PD 98059 on UDP-stimulated ERK1/2 MAPK activation in the absence or presence of inhibitor (50 µM) for 30 min. Data shown are representative of five independent experiments (*<i>p</i><0.05 compared with the UDP control, Student’s <i>t</i>-test). (F) Similar to (A) and (B), treating primary HBE cells with PD 98059 inhibited UDP, but not MRS 2693-stimulated IL-8 release (<i>n</i> = 5–6). The cells were stimulated with the P2Y₆ agonists for 6 h.</p

Measurement of UDP release from 16HBE14o- cells and primary HBE cells damaged by poly-L-arginine.

<p>(A) Probe-1 fluorescence emission intensity as a function of UDP concentration. The line represents a linear regression applied to the data, with a <i>R²</i> of 0.9906. (B) 16HBE14o- cells were incubated in HEPES-buffered saline in the presence of poly-L-arginine (10 µM and 30 µM) for 1 h. Levels of secreted UDP were measured using Probe-1 as described. Each column represents the mean ± S.E. (<i>n</i> = 4). Statistically significant effects compared with vehicle control are marked with an asterisk (*<i>p</i><0.05, Student’s <i>t</i>-test). (C) Similar experiments were performed in primary HBE cells (<i>n</i> = 4).</p

Involvement of p38 MAPK in IL-6 and IL-8 release.

<p>(A, B) Cells were treated with UDP alone or co-incubated with p38 MAPK inhibitor, SB 203580 (50 µM), for 6 h. IL-6 (A) and IL-8 (B) levels were measured with an ELISA. Levels of IL-6 and IL-8 were corrected with vehicle control alone and normalized to UDP-stimulated cytokine release. Each column represents the mean ± S.E. (<i>n</i> = 4–6). Statistically significant inhibitory effects compared with UDP-treated control are marked with an asterisk (*<i>p</i><0.05, Student’s <i>t</i>-test). (C) Effect of SB 203580 (50 µM) alone on basal IL-6 (16HBE14o- cells) or IL-8 (16HBE14o- and primary HBE cells) release. Each column represents the mean ± S.E. (<i>n</i> = 4–6). Statistically significant inhibitory effects compared with untreated control are marked with an asterisk (*<i>p</i><0.05, Student’s <i>t</i>-test). (D) The 16HBE14o- cells were stimulated with 100 µM UDP for the indicated times. Protein matched lysates were analyzed by western blotting with antibodies against phospho-p38 MAPK and p38 MAPK. The graph represents fold change in phosphorylation signal over total MAPK, normalized to GAPDH. The phosphoMAPK∶total MAPK ratio of the vehicle control was arbitrarily set to 1. Data shown are representative of 6–7 independent experiments (*<i>p</i><0.05 compared with vehicle control, Student’s <i>t</i>-test). (E) Similar western blot experiments were performed to show the inhibitory effect of SB 203580 on UDP-stimulated p38 MAPK activation in the absence or presence of the inhibitor (50 µM) for 60 min. Data shown are representative of five independent experiments (*<i>p</i><0.05 compared with the UDP control, Student’s <i>t</i>-test). (F) Similar to (A) and (B), primary HBE cells were stimulated with UDP or MRS 2693 for 6 h in the absence or presence of SB 203580 (<i>n</i> = 5–6).</p

UDP stimulated translocation of NF-κB.

<p>The 16HBE14o- cells were stimulated with vehicle control (A, B) and 100 µM UDP (C, D) for 60 min. The nuclei (A, C) were stained with Hoechst 33342, and NF-<i>κ</i>B (B, D) was stained with anti-NF-<i>κ</i>B and Alexa Fluor 488 goat anti-rabbit secondary antibody (magnification 20×). (E) Immunofluorescence of NF-<i>κ</i>B in the nuclear region and cytoplasmic region was quantified using NIH ImageJ software. Nucleus∶cytoplasm ratios of NF-κB staining were calculated. Each column represents the mean ± S.E. (<i>n</i> = 3) (*<i>p</i><0.01 compared with vehicle control, Student’s <i>t</i>-test).</p

Involvement of P2Y₆ receptors in the poly-L-arginine-induced cytokine response.

<p>Human bronchial epithelial cells (16HBE14o-) were treated with a P2Y₆ receptor antagonist, MRS 2578, alone or with poly-L-arginine for 3 h in the absence or presence of different concentrations of the blocker. Cell-free supernatants were assayed by ELISA for IL-6 (A) and IL-8 (B). Levels of IL-6 and IL-8 were corrected with vehicle control alone and normalized to 3 µM poly-L-arginine. Each column represents the mean ± S.E. (<i>n</i> = 5–7). Statistically significant inhibitory effects compared with poly-L-arginine-treated controls are marked with an asterisk (*<i>p</i><0.05, Student’s <i>t</i>-test). (C-F) Similar experiments were performed by stimulating 16HBE14o- cells with the natural ligand for the P2Y₆ receptor, UDP (10 µM and 100 µM), or a specific P2Y₆ agonist, MRS 2693. Cell supernatants were analyzed for IL-6 (C, E) and IL-8 (D, F). Each column represents the mean ± S.E. (<i>n</i> = 4–8). Statistically significant stimulatory effects compared with vehicle control (normalized to 1) are marked with an asterisk (*<i>p</i><0.05, Student’s <i>t</i>-test). (G, H) UDP or MRS 2693 also stimulated IL-8 release in primary HBE cells (<i>n</i> = 5–7).</p

UDP increased [Ca²⁺]_i, IL-6, and IL-8 mRNA expression.

<p>(A) Concentration-response of UDP upon changes in [Ca²⁺]_i in primary HBE cells. The cells were stimulated with different concentrations of UDP, and the Fura-2 ratio was quantified and plotted against the concentration of UDP (<i>n</i> = 3–7). Each data point represents the mean ± S.E. (B) The effect of 100 µM UDP on [Ca²⁺]_i in 16HBE14o- cells was significantly larger than that of primary HBE cells. (*<i>p</i><0.05, Student’s <i>t</i>-test, <i>n</i> = 8). (C–D) The qRT-PCR analysis of mRNA for IL-6 (C) and IL-8 (D). Epithelia were treated with UDP (100 µM) for 6 h. Relative expression of IL-6 and IL-8 was normalized to GAPDH and is shown as fold change relative to untreated controls. Each column represents the mean ± S.E. (<i>n</i> = 5).</p

Rabbinic Elements in the Verbal System of Maskilic Hebrew Fiction 1857-1881

a Hazard ratio (HR) curve was generated for prosaposin (PSAP) using the TCGA dataset. PSAP expressions between 25 and 75 % quantiles have a HR consistently >1. b Androgen receptor (AR) mRNA does not associate with poor disease-free survival (DFS) in endocrine-treated breast cancer. Kaplan-Meier survival curves were generated to assess the impact of high androgen receptor transcript levels on survival of endocrine-treated patients with breast cancer (n = 661). (PDF 204 kb

Additional file 6: of Prosaposin activates the androgen receptor and potentiates resistance to endocrine treatment in breast cancer

Table: Estrogen receptor (ER) motif results ( p <0.001). A 400-bp-sized window surrounding starting sites of HOXC11 target genes was selected for ER motif searching. The searching process was performed using the FIMO program available in MEME-suite with the p value significant cutoff set at 0.001. (PDF 198 kb

Synthesis of Nitrogen- and Sulfur-Codoped 3D Cubic-Ordered Mesoporous Carbon with Superior Performance in Supercapacitors

In this contribution, nitrogen- and sulfur-codoped 3D cubic-ordered mesoporous carbon (KNOMC) materials with controlled dopant content (10.0–4.6 atom % for nitrogen and 0.94–0.75 atom % for sulfur) are presented, using KIT-6 as the template and pyrrole as the precursor, and its supercapacitive behavior is also investigated. The presented materials exhibit excellent supercapacitive performance by combining electrical double-layer capacitance and pseudocapacitance as well as the enhanced wettability and improved conductivity generated from the incorporation of nitrogen and sulfur into the framework of carbon materials. The specific capacitance of the presented materials reaches 320 F g^–1 at a current density of 1 A g^–1, which is significantly larger than that of the pristine-ordered mesoporous carbon reported in the literature and can even compete with some metal oxides and conducting polymers

Data_Sheet_1_Functional characterization of the Serine acetyltransferase family genes uncovers the diversification and conservation of cysteine biosynthesis in tomato.PDF

Sulfur-containing compounds are essential for plant development and environmental adaptation, and closely related to the flavor and nutrition of the agricultural products. Cysteine, the first organic sulfur-containing molecule generated in plants, is the precursor for most of these active substances. Serine acetyltransferase (SERAT) catalyzes the rate-limiting step of its formation. However, despite their importance, systematic analyses of these enzymes in individual species, especially in economically important crops, are still limited. Here, The SERAT members (SlSERATs, four in total) were identified and characterized in tomato. Phylogenetically, the four SlSERAT proteins were classified into three subgroups with distinct genomic structures and subcellular localizations. On the function, it was interesting to find that SlSERAT3;1, possessed a high ability to catalyze the formation of OAS, even though it contained a long C-terminus. However, it retained the essential C-terminal Ile, which seems to be a characteristic feature of SERAT3 subfamily members in Solanaceae. Besides, SlSERAT1;1 and SlSERAT2;2 also had high activity levels and their catalyzing abilities were significantly improved by the addition of an OAS-(thiol)-lyase protein. At the transcriptional level, the four SlSERAT genes had distinct expression patterns during tomato plant development. Under abiotic stress conditions, the chloroplast-localized SlSERATs were the main responders, and the SlSERATs adopted different strategies to cope with osmotic, ion toxicity and other stresses. Finally, analyses in the loss-of-function and overexpression lines of SlSERAT1;1 suggested that function redundancy existed in the tomato SERAT members, and the tomato SERAT member was ideal target for S-assimilation manipulating in molecular breeding.</p