Clustering and characterization of the differential expression of genes.

<p>(A) SE38 group vs. PBS group. S95H_NS, S50H_NS and S96_H_NS belong to SE38 group; S32H_NS, S32H_2_NS and S71H_NS belong to PBS group. DE genes that showed clear functional annotation at 4 dpi between the SE38 and PBS groups were selected for cluster analysis as described in methods. At 4 dpi, a set of 262 genes were upregulated and the remaining 132 genes were downregulated. (B) G4T10 group vs. PBS group. S98H_NS, SR_1_H_NS and SR_2_H_NS belong to G4T10 group; S32H_NS, S32H_2_NS and S71H_NS belong to PBS group. Each row represents a separate gene and each column represents a separate piglet. Red indicates the increased gene expression levels; green denotes the decreased levels compared with normal samples.</p

Representative histopathological photomicrograph of heart lesions in pigs infected with <i>E</i>. <i>rhusiopathiae</i> strain SE38, G4T10 and control.

<p>(A) Heart of a SE38-infected pig with endocarditis and neutrophil infiltration. (B) Thrombogenesis, myocardial necrosis, and inflammatory cell infiltration. (C) Heart of a G4T10-infected pig at 200 and 50μm. (D) PBS control at 200 and 50 μm.</p

DE genes analysis base on KEGG in SE38 and PBS group.

<p>DE genes analysis base on KEGG in SE38 and PBS group.</p

Transcription analysis of the responses of porcine heart to <i>Erysipelothrix rhusiopathiae</i>

<div><p><i>Erysipelothrix rhusiopathiae</i> (<i>E</i>. <i>rhusiopathiae</i>) is the causative agent of swine erysipelas. This microbe has caused great economic losses in China and in other countries. In this study, high-throughput cDNA microarray assays were employed to evaluate the host responses of porcine heart to <i>E</i>. <i>rhusiopathiae</i> and to gain additional insights into its pathogenesis. A total of 394 DE transcripts were detected in the active virulent <i>E</i>. <i>rhusiopathiae</i> infection group compared with the PBS group at 4 days post-infection. Moreover, 262 transcripts were upregulated and 132 transcripts were downregulated. Differentially expressed genes were involved in many vital functional classes, including inflammatory and immune responses, signal transduction, apoptosis, transport, protein phosphorylation and dephosphorylation, metabolic processes, chemotaxis, cell adhesion, and innate immune responses. Pathway analysis demonstrated that the most significant pathways were Chemokine signaling pathway, NF-kappa B signaling pathway, TLR pathway, CAMs, systemic lupus erythematosus, chemokine signaling pathway, Cytokine–cytokine receptor interaction, PI3K-Akt signaling pathway, Phagosome, HTLV-I infection, Measles, Rheumatoid arthritis and natural-killer-cell-mediated cytotoxicity. The reliability of our microarray data was verified by performing quantitative real-time PCR. This study is the first to document the response of piglet heart to <i>E</i>. <i>rhusiopathiae</i> infection. The observed gene expression profile could help screen potential host agents that can reduce the prevalence of <i>E</i>. <i>rhusiopathiae</i>. The profile might also provide insights into the underlying pathological changes that occur in pigs infected with <i>E</i>. <i>rhusiopathiae</i>.</p></div

STRING analysis of the relationship between DE genes.

<p>(A) the DE genes in piglets infected ER were analyzed using STRING database. (B) the network of DE genes related to TLR4.</p

Results of culture and PCR analysis for three pigs challenged with <i>E</i>. <i>rhusiopathiae</i> presented as the number of positive pigs/total pig samples.

<p>Results of culture and PCR analysis for three pigs challenged with <i>E</i>. <i>rhusiopathiae</i> presented as the number of positive pigs/total pig samples.</p

Immobilization of Antimicrobial Peptide IG-25 onto Fluoropolymers via Fluorous Interactions and Click Chemistry

We report a practical method for biofunctionalization of fluoropolymers based on noncovalent, fluorous interactions and click chemistry that allows incorporation of biomolecules under physiological solutions. We demonstrate the method by immobilization of an antimicrobial peptide (AMP) on fluorous thin films and fluorosilicone contact lens. The fluorous surfaces were dip-coated with fluorous-tagged oligo­(ethylene) chain terminated with a reactive group, such as an alkynyl group. This simple step generates a “clickable” surface. The noncovalent fluorous interaction was strong enough to allow subsequent covalent attachment of IG-25, a truncated version of the most extensively studied human AMP LL-37. The attachment was through copper-catalyzed click reaction between the alkynyl group on the surface and the azido-OEG tag at the N-terminus of IG-25. In comparison to surfaces presenting IG-25 randomly bound via carbodiimide chemistry, the surfaces presenting IG-25 tethering to the surface at the <i>N</i>-terminus via click chemistry displayed higher antibacterial activities against an ocular pathogen <i>Pseudomonas aeruginosa</i> (strain PA-O1)

Gold Nanoshell-Decorated Silicone Surfaces for the Near-Infrared (NIR) Photothermal Destruction of the Pathogenic Bacterium E. faecalis

Catheter-related infections (CRIs) are associated with the formation of pathogenic biofilms on the surfaces of silicone catheters, which are ubiquitous in medicine. These biofilms provide protection against antimicrobial agents and facilitate the development of bacterial resistance to antibiotics. The application of photothermal agents on catheter surfaces is an innovative approach to overcoming biofilm-generated CRIs. Gold nanoshells (AuNSs) represent a promising photothermal tool, because they can be used to generate heat upon exposure to near-infrared (NIR) radiation, are biologically inert at physiological temperatures, and can be engineered for the photothermal ablation of cells and tissue. In this study, AuNSs functionalized with carboxylate-terminated organosulfur ligands were attached to model catheter surfaces and tested for their effectiveness at killing adhered Enterococcus faecalis (E. faecalis) bacteria. The morphology of the AuNSs was characterized by scanning electron microscopy (SEM) and transmission electron microscopy (TEM), while the elemental composition was characterized by energy-dispersive X-ray spectroscopy (EDX) and X-ray photoelectron spectroscopy (XPS). Furthermore, optical and photothermal properties were acquired by ultraviolet–visible (UV-vis) spectroscopy and thermographic imaging with an infrared camera, respectively. Bacterial survival studies on AuNS-modified surfaces irradiated with and without NIR light were evaluated using a colony-formation assay. These studies demonstrated that AuNS-modified surfaces, when illuminated with NIR light, can effectively kill E. faecalis on silicone surfaces

“Click” Immobilization of a VEGF-Mimetic Peptide on Decellularized Endothelial Extracellular Matrix to Enhance Angiogenesis

We show that coating of decellularized extracellular matrix (DC-ECM) on substrate surfaces is an efficient way to generate a platform mimicking the native ECM environment. Moreover, the DC-ECM can be modified with a peptide (QK) mimicking vascular endothelial growth factor without apparently compromising its integrity. The modification was achieved through metabolic incorporation of a “clickable” handle to DC-ECM followed by rapid attachment of the QK peptide with an azido tag using copper-catalyzed click reaction. The attachment of the QK peptide on to DC-ECM in this way further enhanced the angiogenic responses (formation of branched tubular networks) of endothelial cells