Journal: Brain : a journal of neurology

Article Title: Connexin43 mimetic peptide reduces vascular leak and retinal ganglion cell death following retinal ischaemia.

doi: 10.1093/brain/awr338

Figure Lengend Snippet: Figure 3 Three hours of hypoxia and 6 h of reperfusion lead to significant endothelial cell death in vitro in rat brain microvascular endothelial cells (RBMVEC; A). Non-specific gap junction blocker carbenoxolone, non-specific hemichannel blocker LaCl3, and connexin43 (Cx43) mimetic peptide protected endothelial cells against hypoxic injury, with the number of viable cells significantly higher than no treatment, while scrambled peptide did not have any protective effects. The number of viable cells was expressed as percentage of the control without hypoxia. Hypoxia and reperfusion also lead to significant propidium iodide dye uptake into primary rat brain microvascular endothelial cells indicating open hemichannels (B). Carbenoxolone, LaCl3 and connexin43 mimetic peptide significantly prevented dye uptake compared to no treatment, indicating hemichannel closure, while scrambled peptide did not have any effect. Stars denote statistical significance when compared to the control group or compared between groups in brackets; P 5 0.05.

Article Snippet: Rat brain microvascular endothelial cells (R840K-05a, Cell Applications) were plated into 24-well plates (1 105 cells/well) in rat brain microvascular endothelial cell growth media (R819K-500, Cell Applications) and allowed to settle for 16 h. Medium was then removed and replaced with Dulbecco’s Modified Eagle’s Medium/F12 containing 0.5% foetal bovine serum and 1% glutamine.

Techniques: In Vitro, Control

Journal: Experimental Animals

Article Title: Experimental transmission of systemic AA amyloidosis in autoimmune disease and type 2 diabetes mellitus model mice

doi: 10.1538/expanim.16-0037

Figure Lengend Snippet: Amyloid deposits in spleens of several mice. Amyloid deposits were stained with Congo red and observed as green birefringence under polarized light. Amyloid fibrils in spleens of 18-week-old mice (C57BL, C57BL lpr/lpr , C57BL Ham +/+, and C57BL ob/ob ) inoculated with murine amyloid fibrils were reacted with AF2948 antibody in IHC. Also, amyloid fibrils in spleens of 18-and 30-week-old C57BL Ham +/+ mice inoculated with bovine amyloid fibrils and chicken amyloid fibrils, respectively, were reacted with AF2948 antibody in IHC. Bar=100 µ m.

Article Snippet: The membranes were incubated with anti-mouse SAA1 goat antibody (1:5,000) (AF2948, R&D Systems, Minneapolis, MN, USA), anti-bovine SAA1 monoclonal antibody, 25BF12 (1:1,600) [ ], which was purified from the culture supernatant of hybridoma cells by protein A-sepharose (GE Healthcare), or anti-chicken SAA1 mouse antibody (1:1,600) [ ] in 1% skim milk in PBST at room temperature for 1 h. Membranes were washed three times with PBST and incubated with anti-goat IgG donkey antibody F (ab’) 2 conjugated with horseradish peroxidase (HRP) (1:1,000) (SC3851, Santa Cruz Biotechnology, Dallas, TX, USA) , anti-mouse IgG F (ab’) 2 conjugated with HRP (1:1,600) (NA9310V, GE Healthcare, Buckinghamshire, UK) in 1% skim milk in PBST at room temperature for 45 min.

Techniques: Staining

Journal: Experimental Animals

Article Title: Experimental transmission of systemic AA amyloidosis in autoimmune disease and type 2 diabetes mellitus model mice

doi: 10.1538/expanim.16-0037

Figure Lengend Snippet: Intensity scoring of amyloid deposits in the spleen using ImageJ. Amyloid fibrils in spleens of mice inoculated with murine, bovine, or chicken amyloid fibrils were reacted with AF2948 antibody in IHC. Images of IHC indicate amyloid deposits around spleen follicles. ImageJ was used to measure the intensity of amyloid deposits. ImageJ intensity scores are shown at the bottom. Bar=400 µ m.

Article Snippet: The membranes were incubated with anti-mouse SAA1 goat antibody (1:5,000) (AF2948, R&D Systems, Minneapolis, MN, USA), anti-bovine SAA1 monoclonal antibody, 25BF12 (1:1,600) [ ], which was purified from the culture supernatant of hybridoma cells by protein A-sepharose (GE Healthcare), or anti-chicken SAA1 mouse antibody (1:1,600) [ ] in 1% skim milk in PBST at room temperature for 1 h. Membranes were washed three times with PBST and incubated with anti-goat IgG donkey antibody F (ab’) 2 conjugated with horseradish peroxidase (HRP) (1:1,000) (SC3851, Santa Cruz Biotechnology, Dallas, TX, USA) , anti-mouse IgG F (ab’) 2 conjugated with HRP (1:1,600) (NA9310V, GE Healthcare, Buckinghamshire, UK) in 1% skim milk in PBST at room temperature for 45 min.

Techniques: