Additional file 1: Fiugre S1. of HECTD1 controls the protein level of IQGAP1 to regulate the dynamics of adhesive structures

Generation of Hectd1 mutant animals. (PPTX 142 kb

Additional file 5: Figure S4. of HECTD1 controls the protein level of IQGAP1 to regulate the dynamics of adhesive structures

Loss of HECTD1 leads to mislocalization of α-actinin and paxillin/zyxin. (PPTX 1571 kb)

In Vivo Delivery of Adenoviral Vector Containing Interleukin-17 Receptor A Reduces Cardiac Remodeling and Improves Myocardial Function in Viral Myocarditis Leading to Dilated Cardiomyopathy

<div><p>Th17 cells have been implicated in the pathogenesis of myocarditis. Interleukin (IL)-17A produced by Th17 cells is dispensable for viral myocarditis but essential for the progression to dilated cardiomyopathy (DCM). This study investigated whether the adenoviral transfer of the IL-17 receptor A reduces myocardial remodeling and dysfunction in viral myocarditis leading to DCM. In a mouse model of Coxsackievirus B3 (CVB3)-induced chronic myocarditis, the delivery of the adenovirus-containing IL-17 receptor A (Ad-IL17RA:Fc) reduced IL-17A production and decreased the number of Th17 cells in the spleen and heart, leading to the down-regulation of systemic TNF-α and IL-6 production. Cardiac function improved significantly in the Ad-IL17R:Fc- compared with the Ad-null-treated mice 3 months after the first CVB3 infection. Ad-IL17R:Fc reduced the left ventricle dilation and decreased the mortality in viral myocarditis, leading to DCM (56% in the Ad-IL17R:Fc versus 76% in the Ad-null group). The protective effects of Ad-IL17R-Fc on remodeling correlated with the attenuation of myocardial collagen deposition and the reduction of fibroblasts in CVB3-infected hearts, which was accompanied by the down-regulation of A distintegrin and metalloprotease with thrombospondin type 1 motifs (ADAMTS-1), Matrix metalloproteinase-2(MMP-2), and collagen subtypes I and III in the heart. Moreover, in cultured cardiac fibroblasts, IL-17A induced the expression of ADAMTS-1, MMP-2, and collagen subtypes I and III and increased the proliferation of fibroblasts. We determined that the delivery of IL-17-RA:Fc reduces cardiac remodeling, improves function, and decreases mortality in viral myocarditis leading to DCM, possibly by suppressing fibrosis. Therefore, the adenoviral transfer of the IL-17 receptor A may represent an alternative therapy for chronic viral myocarditis and its progression to DCM.</p></div

Recovery of Ammonium and Phosphate from Wastewater by Wheat Straw-based Amphoteric Adsorbent and Reusing as a Multifunctional Slow-Release Compound Fertilizer

To minimize the negative impact of nitrogen and phosphorus pollution from wastewater and improve fertilizer use efficiency, a novel multifunctional slow-release compound fertilizer was prepared by recovery of NH₄⁺ and H₂PO₄^– from aqueous solutions onto amphoteric straw cellulose (ASC) adsorbent. The effects of adsorbent dosage, solution pH, contact time and ionic strength on adsorption were investigated. The adsorption quickly reached equilibrium. The maximum NH₄⁺ adsorption capacity of ASC was 68.4 mg g^–1 at around pH 7.0, whereas it was 38.6 mg g^–1 for H₂PO₄^– adsorption at pH 5.0. Moreover, the feasibility of reusing the nutrients-laden carrier material as a multifunctional slow-release compound fertilizer was determined. The study demonstrated the product with excellent slow-release and water-retention properties. Thus, it could improve soil moisture content and reduce soil moisture evaporation rate and was economical and environmentally friendly for application in horticulture and agriculture

Ad-IL-17AR:Fc administration reduces IL-17A production in the blood and the heart.

<p>(<i>A</i>) Serum IL-6 concentrations at post-infection 7 days, 2mons and 3mons in PBS-, Ad-IL-17AR:Fc- and Ad:null-treated mice (n = 5) were measured using ELISA, and the data were analyzed by a 2-factor ANOVA. ^##<i>p</i><0.01, ^#<i>p</i><0.05 vs 7 Days group, **<i>p</i><0.01, *<i>p</i><0.05 vs control group, (<i>B</i>) Serum TNF-α concentrations at post-infection 7 days, 2mons and 3mons in PBS-, AdIL-17R:Fc- and Ad:null-treated mice (n = 5) were measured using ELISA, and the data were analyzed by a 2-factor ANOVA. ^#<i>p</i><0.05 vs 7 Days group, *<i>p</i><0.05 vs control group. The data represent the mean ± SEM for 6 mice per group. (<i>C</i>)The IL-17A protein extracted from the left ventricle of the mice in the three groups at post-infection day 90, was subjected to western blotting and was probed with the indicated antibodies (n = 3). (D)The IL-17A protein level is expressed as a ratio of the GAPDH level in the same sample; the data are expressed as the mean± SEM. *<i>P</i><0.05 vs the control group. (<i>E</i>) The IL-17RA protein extracted from the left ventricle of the mice in the three groups at post-infection day 90, was subjected to western blotting and was probed with the indicated antibodies (n = 3). (<i>F</i>) The IL-17RA protein level is expressed as a ratio of the GAPDH level in the same sample; the data are expressed as the mean± SEM.</p

The levels of IL-6 and TNF-α in the supernatants of cardiac fibroblast cultures treated with IL-17.

<p>The IL-17 (10 ng/ml)-treated group produced higher levels of IL-6 and TNF-α cytokines than the control- and IL-17 (5 ng/ml)-treated groups. Moreover, IL-6 in the IL-17 (5 ng/ml)-treated group was highly expressed in the supernatants of the heart homogenates (ELISA). The values in the IL-17-treated CFs were normalized to the values in the control cells (n = 5 per group). The data represent the mean ± SEM. **<i>P</i><0.01 vs control group, ^##<i>P</i><0.01 vs control group.</p

Degradation of CFs related to the MMPs/TIMPs expression following neutralization of IL-17.

<p>(<i>A</i>) Representative western blots showing the expression of the cardiac ADAMTS-1 protein in each group. (<i>B</i>) Representative western blots showing the expression of the cardiac TIMP-1 protein in each group. (<i>C</i>) Representative western blots showing the expression of the cardiac MMP-2 protein in each group. The expression of the cardiac ADAMTS-1 and MMP-2/TIMP-1 proteins in each group is expressed as a ratio of the GAPDH expression for the same sample. On day 90, 3 mice per group were analyzed. *<i>P</i><0.05 vs control group.</p

The number of Th17 cells in the spleen and cardiac tissue is decreased by Ad-IL-17AR:Fc.

<p>(<i>A</i>) Representative flow cytometry images of Th17 (CD4⁺ IL-17⁺) cells from each group gated on CD4⁺ T cells. Numbers in the upper right quadrants and lower right quadrants indicate the separate percentages of Th17 cells and CD4⁺ T cells, respectively. PE = phycoerythrin; FITC = fluorescein isothiocyanate. (<i>B</i>) The percentage of Th17 (CD4⁺ IL-17⁺) cells in each group was analyzed using CellQuest software. (<i>C</i>) The levels of IL-6 and TNF-α in the supernatants of heart homogenates detected using ELISA. The data represent the mean ± SEM for 6 mice per group. **<i>P</i><0.01 vs control group, *<i>p</i><0.05 vs control group.</p

IL-17-stimulated MMPs/TIMPs protein expression, collagen deposition and proliferation in CFs.

<p>(<i>A</i>) Representative western blot for ADAMTS-1 protein levels in CFs treated with different doses (0, 10, and 50 ng/ml) of IL-17 for 24 h, and at 0, 24 and 48 h. (<i>B</i>) IL-17-mediated expression of ADAMTS-1 and MMP-2/TIMP-1 in the CFs treated with IL-17 (10 ng/ml) for 24 h, using TGF-β (10 ng/ml) as a control. (<i>C</i>) RT-PCR in CFs showing the IL-17-induced expression of COI1α2 and COI3α1 mRNAs. (<i>D</i>) The proliferation of CFs following the administration of different doses of IL-17 using cell counting kit-8 (CCK-8). (<i>E</i>) Western blot showing the expression of DDR2 at the different concentrations of IL-17. (<i>F</i>) Immunofluorescence in the cultured CFs showing the expression of the DDR2 protein in the 10 ng/ml IL-17, 5 ng/ml IL-17 and control groups (magnification ×200). The values in the IL-17-treated CFs were normalized to the values in the control cells (n = 5 per group). The data represent the mean ± SEM. *<i>P</i><0.05 vs control group, **<i>P</i><0.01 vs control group.</p