Journal:

Article Title: Improved outcome in shigellosis associated with butyrate induction of an endogenous peptide antibiotic

doi: 10.1073/pnas.0602888103

Figure Lengend Snippet: Immunohistochemical staining of CAP-18 in paraffin-embedded sections of rabbit colon. All sections were counterstained with hematoxylin. (A) Immunoreactive signals for CAP-18 (brown) in healthy rabbits were almost exclusively located in the SE. (B) In Shigella-infected rabbits, surface and crypt epithelia were almost devoid of CAP-18 staining (arrowheads); abundant CAP-18-expressing inflammatory cells (arrows) were seen in the LP. (C) Reappearance of CAP-18 staining in the SE and disappearance of CAP-18-expressing cells from the LP in infected rabbits treated with butyrate.

Article Snippet: Polyclonal antiserum against CAP-18 was made in chicken (Innovagen), and enriched IgY was purified by affinity chromatography, where the CAP-18 peptide was crosslinked to the gel matrix (Innovagen).

Techniques: Immunohistochemical staining, Staining, Infection, Expressing

Journal:

Article Title: Improved outcome in shigellosis associated with butyrate induction of an endogenous peptide antibiotic

doi: 10.1073/pnas.0602888103

Figure Lengend Snippet: The relative expression of CAP-18 transcript in colonic biopsies from healthy, Shigella-infected, and butyrate-treated rabbits was measured by quantitative real-time PCR. Each bar represents data from one rabbit. For each rabbit, the first bar represents proximal colon, and the second bar represents distal colon. The last bar represents distal colon from only one rabbit. Under each bar, histological grades of inflammation are expressed as mild, 1; moderate, 2; severe, 3; and normal, 0 histology.

Article Snippet: Polyclonal antiserum against CAP-18 was made in chicken (Innovagen), and enriched IgY was purified by affinity chromatography, where the CAP-18 peptide was crosslinked to the gel matrix (Innovagen).

Techniques: Expressing, Infection, Real-time Polymerase Chain Reaction

Journal:

Article Title: Improved outcome in shigellosis associated with butyrate induction of an endogenous peptide antibiotic

doi: 10.1073/pnas.0602888103

Figure Lengend Snippet: Western blot analysis of CAP-18 in stool extracts from healthy, Shigella-infected and butyrate-treated rabbits with the CAP-18-specific polyclonal antisera. Lanes 1 and 2 show a marker and the synthetic CAP-18 peptide, respectively. A single band corresponding to the pro-CAP-18 protein appears in the stool of healthy (lanes 3 and 4) and Shigella-infected (lane 5) rabbits 1 day postinfection. Two immunoreactive bands are visualized at day 2 postinfection (lane 6), showing presence of both pro-CAP-18 and active CAP-18 peptides in stool. Butyrate treatment resulted in release of the active CAP-18 peptide in stool (lanes 7–10); day-1 and day-2 stool specimens (lanes 7 and 8) also contained the pro-CAP-18 protein.

Article Snippet: Polyclonal antiserum against CAP-18 was made in chicken (Innovagen), and enriched IgY was purified by affinity chromatography, where the CAP-18 peptide was crosslinked to the gel matrix (Innovagen).

Techniques: Western Blot, Infection, Marker

Journal:

Article Title: Improved outcome in shigellosis associated with butyrate induction of an endogenous peptide antibiotic

doi: 10.1073/pnas.0602888103

Figure Lengend Snippet: Inhibitory effects of various concentration of CAP-18 in presence (dotted line) or absence (solid line) of 40 mM butyrate (4.4 mg/ml) on the growth of Shigella bacterial colonies.

Article Snippet: Polyclonal antiserum against CAP-18 was made in chicken (Innovagen), and enriched IgY was purified by affinity chromatography, where the CAP-18 peptide was crosslinked to the gel matrix (Innovagen).

Techniques: Concentration Assay

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Treatment with Entinostat Heals Experimental Cholera by Affecting Physical and Chemical Barrier Functions of Intestinal Epithelia

doi: 10.1128/AAC.02570-16

Figure Lengend Snippet: CAP-18 peptide/protein expression in the ileal mucosae of healthy rabbits, Vibrio cholerae-infected rabbits, and rabbits that were infected and treated with entinostat. Mucosal sections of ileum from healthy rabbits (n = 5), V. cholerae-infected rabbits (n = 5), and infected rabbits treated with 0.5 mg entinostat twice a day for 2 days (n = 5) or with 1 mg entinostat once a day for 2 days (n = 5) were stained with a CAP-18-specific antibody. (A) Representative photomicrographs. CAP-18-positive staining is brown; cell nuclei are stained blue with hematoxylin. (Top left) Prominent expression of CAP-18 (thick arrows) in the SE of healthy ileal mucosae. (Top right) Downregulation of CAP-18 expression in the SE after V. cholerae infection. Partial erosion of SE (arrowheads) was also noted. (Bottom) Recovery of epithelial erosion and restoration of CAP-18 expression (arrows) in the SE after treatment with 0.5 mg entinostat twice a day for 2 days (left) or 1 mg entinostat once a day for 2 days (right). Bars, 50 μm (top and bottom left); 100 μm (bottom right). (B) Semiquantitative expression of CAP-18 in SE, given as ACIA scores (see Materials and Methods). Data are means ± SEM. Differences in outcomes between different groups were estimated by mixed-model ANOVA. Differences are significant at a P value of <0.05.

Article Snippet: The affinity-purified polyclonal chicken antiserum against CAP-18 (Innovagen) recognizes both the proform and the mature CAP-18 peptide.

Techniques: Expressing, Infection, Staining

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Treatment with Entinostat Heals Experimental Cholera by Affecting Physical and Chemical Barrier Functions of Intestinal Epithelia

doi: 10.1128/AAC.02570-16

Figure Lengend Snippet: In vitro inhibitory effect of entinostat or CAP-18 on the growth of Vibrio cholerae. A suspension of V. cholerae Ogawa was incubated with different concentrations of entinostat (A) or synthetic CAP-18 (B) in MHB in wells of a microtiter plate at 37°C for 2 h. Bacterial suspensions from individual wells were plated on TTGA, and CFU were counted after overnight incubation. Data are means ± SEM from 2 and 4 individual experiments for entinostat and CAP-18, respectively.

Article Snippet: The affinity-purified polyclonal chicken antiserum against CAP-18 (Innovagen) recognizes both the proform and the mature CAP-18 peptide.

Techniques: In Vitro, Incubation