Journal: Redox Biology

Article Title: The nitroxide 4-methoxy-tempo inhibits the pathogenesis of dextran sodium sulfate-stimulated experimental colitis

doi: 10.1016/j.redox.2019.101333

Figure Lengend Snippet: MetT attenuates body weight loss, and clinical and histopathological parameters in mice challenged with DSS in the drinking water. Mice exposed to normal water or 3% w/v DSS were treated with vehicle or MetT (15 mg/kg), twice daily by i. p. injection over 9 days. Panel (a) Percent body weight loss in mice over 9-days DSS insult. Data is expressed as the percentage of the original weight prior to treatment. Panel (b) Enumerated clinical score representing reduced mobility, faecal consistency and rectal bleeding in mice at day 9 of DSS challenge. Panel (c) Pattern-based recognition of intact crypts (InForm V2.1.1), as represented by percent fractional area of the total colonic mucosa area in the same colon section following sacrifice at day 9 of DSS challenge. Crypt drop out was assessed using a histological Alcian blue stain for mucin after 9 days of DSS-insult. Panel (d) Representative Alcian blue and Safranin O staining in control mice and following DSS-induced colitis with and without MetT treatment. Panel (e) Software-based quantification (MetaMorph® V7.8) of the extent of Alcian blue staining included the transverse and descending regions of the colon that was subsequently normalised to the combined length of the aforementioned regions. Slides were imaged with a Zeiss Axio Scan. Z1 slide scanner and pseudo-fields of view were obtained at 1x magnification. Data represents mean ± SD; n = 6 mice per group and corresponding n = 6 data points except for (c), where n > 70 based upon the analysis of each field of view at 20x magnification. Different to vehicle, where * p < 0.05, ** p < 0.005 & *** p < 0.001. Different to DSS group, where # p < 0.05, ## p < 0.005 & ### p < 0.001. n. a. = not applicable. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Panel (e) shows the quantification of 3-NO 2 -Tyr + labelling performed using the software, MetaMorph® V7.8, with 3-NO 2 -Tyr + labelling normalised to the length of colon in each field of view and presented as a fold-change relative to the vehicle group.

Techniques: Injection, Staining, Software

Journal: Redox Biology

Article Title: The nitroxide 4-methoxy-tempo inhibits the pathogenesis of dextran sodium sulfate-stimulated experimental colitis

doi: 10.1016/j.redox.2019.101333

Figure Lengend Snippet: Biomarkers of MPO-mediated oxidative damage after 9 days of DSS insult. Panels represent (a) glutathione sulfonamide and (b) 3-chloro-tyrosine (3-Cl-Tyr); biomarkers were determined in colon homogenates with LC mass spectrometry analysis, while the lipid (per)oxidation biomarker (c) lipid hydroperoxide (LOOH) was assessed using a modified FOX-1 assay from fresh plasma isolated on day 9 of DSS. Panel (d) shows 3-NO 2 -Tyr labelling determined with frozen colon tissue sections immunohistochemically labelled using TSA amplification and juxtaposed to H&E stained serial sections. Arrow indicates 3-NO 2 -Tyr + labelling (red-fluorescent staining). Panel (e) shows the quantification of 3-NO 2 -Tyr + labelling performed using the software, MetaMorph® V7.8, with 3-NO 2 -Tyr + labelling normalised to the length of colon in each field of view and presented as a fold-change relative to the vehicle group. Data represents approximately n = 30 fields of view per mouse with data averaged and plotted as mean ± SD; n = 6 mice per group. Data for glutathione sulfonamide and 3-Cl-Tyr data represent mean ± SD; n = 6 mice per group. Different to corresponding controls; * p < 0.05 and ** p < 0.01. Different to the DSS group in the absence of MetT; # p < 0.05 and ## p < 0.01. Images were digitally magnified from original 20x magnification. Scale bar = 150 μm. (For interpretation of the references to colour in this figure legend, the reader is referred to the Web version of this article.)

Article Snippet: Panel (e) shows the quantification of 3-NO 2 -Tyr + labelling performed using the software, MetaMorph® V7.8, with 3-NO 2 -Tyr + labelling normalised to the length of colon in each field of view and presented as a fold-change relative to the vehicle group.

Techniques: Mass Spectrometry, Biomarker Assay, Modification, Isolation, Amplification, Staining, Software

Journal: Redox Biology

Article Title: The nitroxide 4-methoxy-tempo inhibits the pathogenesis of dextran sodium sulfate-stimulated experimental colitis

doi: 10.1016/j.redox.2019.101333

Figure Lengend Snippet: Representative images and quantification of TUNEL positivity in the colon mucosa following 9 days of DSS challenge. Panels show (a) Representative TUNEL + labelling of double stranded DNA in the colon mucosa, (b) quantification of TUNEL + labelling as performed using the software, MetaMorph® V7.8, with total counts normalised to the total length of the same colon expressed in millimetres. Data is averaged and plotted as mean ± SD; n = 6 mice per group. n = 28 field of view (FOV) averaged from 6 mice (H 2 O + V), n = 15 FOV averaged from 3 mice (DSS + V) and n = 30 FOV averaged from 6 mice (DSS + MetT). Different to DSS group; *** p < 0.001. Different to vehicle group (in the absence of MetT); ### p < 0.001. Images were digitally magnified from original 20x magnification. Scale bar = 200 μm.

Article Snippet: Panel (e) shows the quantification of 3-NO 2 -Tyr + labelling performed using the software, MetaMorph® V7.8, with 3-NO 2 -Tyr + labelling normalised to the length of colon in each field of view and presented as a fold-change relative to the vehicle group.

Techniques: TUNEL Assay, Software