Quantification of atherogenesis <i>en face</i> (A) and in the aortic arch (B) in the mouse model of TGF-ß1 overexpressing macrophages after 8 weeks (8 w), 16 weeks (16 w) and 24 weeks (24 w) on the WTD, respectively.

<p><b>A, left panel</b> Box and whisker diagrams (median, interquartile range, minimum, and maximum; n.s.  =  not significant) of aortic plaque area (%) of SRA-TGF-ß1 ApoE^−/− (grey) and ApoE^−/− mice (white). <b>A, right panel</b> Representative Sudan-stained aortas <i>en face</i> of SRA-TGF-ß1 ApoE^−/− and ApoE^−/− mice. <b>B, right panel</b> Hearts of SRA-TGF-ß1 ApoE^−/− and and ApoE^−/− mice were resected, and measurement of plaque size in longitudinal sections of the aortic arch stained with trichrome was performed as follows: a 2-mm segment of the lesser curvature of the aortic arch was defined proximally by a perpendicular axis dropped from the right side of the innominate artery origin (dashed line) and the aortic-arch wall area subtended by this 2-mm stretch of intima (green line) was calculated for each section of all mice by computerized image analysis. In addition, the aortic-arch intima thickness (red line) was determined on this same segment of the lesser curvature <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0040990#pone.0040990-Blobe1" target="_blank">[1]</a>. IA, innominate artery; LCCA, left common carotid artery; LSA, left subclavian artery. Note, that no statistically significant differences of lesion area could be detected between SRA-TGF-ß1 ApoE^−/− (black circles) and ApoE^−/− (white circles) mice (non-parametric Mann-Whitney U test, n.s.  =  not significant, <b>B, left panel).</b></p

Phenotypic analysis of atherosclerotic lesions in SRA-TGF-ß1 ApoE^−/− mice.

<p><b>A</b> Box and whisker diagrams (median, interquartile range, minimum, and maximum; n.s.  =  not significant) of the quantification of macrophages (upper panel), SMCs (middle panel) and collagen (lower panel) in atherosclerotic lesions of SRA-TGF-ß1 ApoE^−/− females and isogenic ApoE^−/− controls after 8, 16 and 24 weeks on the WTD, respectively. <b>B</b> Representative histological slides of atherosclerotic lesions located in the inner aortic arch intima (lesser curvature) of SRA-TGF-ß1 ApoE^−/− mutants and ApoE^−/− controls after 24 weeks on the WTD. The slides have been stained for macrophages, SMCs, and collagen by using a rat anti-mouse F4/80 antibody (upper panel), a mouse anti-smooth muscle α-actin antibody (middle panel), and picrosirius red with subsequent polarization (lower panel). Percent-positive area for macrophages (upper panels, asterisks), SMCs (middle panels, brown-stained areas), and collagen (lower panels, areas with yellow, green, orange, or red polarized colour) were quantified by Photoshop-based image analysis. The aortic lumen is to the upper left corner. The demarcation between intima and media is indicated by black arrowheads. In the lower panel, note that the adventitial tissue (asterisk) also polarizes after picrosirius red staining (internal positive control).</p

A Generation and expression analysis of transgenic mice with macrophage-specific TGF-ß1 overexpression.

<p>Comparative quantitative RT-PCR analyses was performed with total lung RNA isolated using Tri-Reagent (Sigma-Aldrich, Taufkirchen, Germany) of at least three 8 weeks old transgenic mice of strains SRA-TGF-ß1-B (B), SRA-TGF-ß1-J (J), SRA-TGF-ß1-L (L) and SRA-TGF-ß1-H (H) (left panel). Strain SRA-TGF-ß1-L (L) was used for additional comparative quantitative RT-PCR analysis of total liver RNA (right panel). In both panels, HPRT was used as housekeeping gene for the normalization of the expression data. The relative quantification of the transcripts was done by the 2^(-ΔΔCt) method. <b>B</b> Demonstration of TGF-ß1 expression by immunohistochemistry in both lung (left panel) and liver tissue (right panel) by using a polyclonal goat anti-TGFß1 antibody. <b>C</b> Group size, body weights, serum cholesterol and serum triglyceride concentrations of SRA-TGF-ß1 ApoE^−/− and ApoE^−/− mice. Data are presented as means ± standard deviation.</p

Early fungal growth control in pulmonary infection with <i>C. neoformans</i> in the presence of IL-4Rα signaling.

<p>Wild-type (WT, open circle) and IL-4Rα^−/− (gray circle) mice on C57BL/6J background were infected intranasally with <i>C. neoformans</i>. Analysis of fungal burdens in the lung was done at different days <i>post infectionem</i> (dpi) as indicated. Shown is data from n = 7 mice per group from one representative of three independent experiments (14 dpi) or from two independent experiments (7; 21 and 42 dpi). Statistical analysis was done using the unpaired Student's t-test (7 dpi) or Mann-Whitney test. *P<0.05; **P<0.01.</p

Representative examples of TGFß1 expression in atherosclerotic lesions of SRA-TGF-ß1 ApoE^−/− mice after 24 weeks on the WTD:

<p>Atherosclerotic lesions of the inner aortic arch intima (lesser curvature) were stained with trichrome (upper left panel), rat anti-mouse F4/80 (upper right) for macrophages, mouse anti-smooth muscle α-actin (1A4) (lower left panel) and goat anti-human TGFß1 (lower right panel). The lumen is to the upper left corner. The demarcation between intima and media is indicated by an arrowhead.</p

Stronger pulmonary inflammation, eosinophilia, and mucus production in WT as compared with IL-4Rα^−/− mice.

<p>Lung slices from WT and IL-4Rα^−/− mice infected for 14 days were stained with H&E (A-F) and periodic acid Schiff reagent (G, H). Leukocyte infiltration and fungal load are depicted in panels A, C and B, D. Sites of inflammation contain eosinophils (arrowheads) and large, multinucleated macrophages (E) or lymphocytes (F). Mucus production by bronchial epithelial cells is depicted in G and H. One representative experiment out of three with n = 6–7 animals per group is shown.</p

In the presence of IL-4Rα elevated pulmonary chemokine expression, IFN-γ mRNA expression and NO production.

<p>Following infection, WT (open circle) and IL-4Rα^−/− (gray circle) mice were sacrificed at the time points indicated (A, C) or at 14 days after infection (B). RT-qPCR analyses were done to determine the expression of mRNAs as indicated. Data are derived from one (A, C, 0 and 7 dpi) or one representative out of two (A, 14 dpi) or three (C, 14 dpi) experiments (n = 6–7 mice per genotype and experiment). Statistical analysis was done using the unpaired Student's t-test (ns, not significant; *P<0.05; **P<0.01; ***P<0.001 (A, C)). The concentration of nitric oxide (NO) in cell culture supernatants was determined using the Griess reaction. Pooled data from two different experiments are shown. Dotted line represents detection limit. Statistical analysis was done using the Mann-Whitney test. **P<0.01 (B).</p

Additional file 1 of Tissue S100/calgranulin expression and blood neutrophil-to-lymphocyte ratio (NLR) in dogs with lower urinary tract urothelial carcinoma

Additional file 1: Supplementary Table 1. Overview of the patient demographic, treatment, outcome, and study participation information for the dogs in-cluded in the study (n=55)