Journal: British Journal of Nutrition

Article Title: The human milk oligosaccharide 2′-fucosyllactose attenuates the severity of experimental necrotising enterocolitis by enhancing mesenteric perfusion in the neonatal intestine

doi: 10.1017/s0007114516002944

Figure Lengend Snippet: Fig. 4. Formula supplementation with the human milk oligosaccharide (HMO) 2′-fucosyllactose enhances mesenteric perfusion in experimental necrotising enterocolitis (NEC) via maintenance of intestinal endothelial nitric oxide synthase (eNOS) expression. (a) Representative confocal micrographs of terminal ileal whole mounts from wild-type neonatal mice from the indicated treatment groups: breast-fed controls (BF), breast-fed with HMO (BF + HMO, experimental NEC, formula fed (FF) and FF with HMO (FF + HMO), as well as mice that in addition to the experimental NEC model and HMO treatment also received the eNOS inhibitors N5-(1-iminoethyl)-L-ornithine dihydrochloride (L-NIO) (FF + HMO + L-NIO) or diphenyleneiodonium chloride (DPI) (FF + HMO + DPI) with their formula. After intra- cardiac injection with the fluorescent tracer tomato lectin, which correlates with intestinal blood flow (green), whole mounts were immunostained for platelet endothelial cell adhesion molecule 1 (PECAM-1) to assess the intestinal microvasculature (red). (b) Graph representing villus perfusion index as described in the Materials and methods section expressed as a percentage. (c) Intestinal eNOS mRNA expression by quantitative real-time PCR (qRT-PCR) in the indicated treatment groups relative to the housekeeping gene RPLO. (d) SDS-PAGE immunoprecipitation Western blot (IP-WB) performed on the terminal ileum of wild-type mice in the indicated treatment groups. IgG is shown as a loading control (i); densitometry quantification with Image J performed on three samples per group in (ii). * P < 0·05 v. breast-fed control animals, ** P < 0·05 v. FF. Values are means with their standard errors. Scale bar is 50 μm. Representative of at least three separate experiments, where each mouse represents an individual symbol on a graph.

Article Snippet: The fluorescein-labelled Lycopersicon esculentum (tomato) lectin (Vector Laboratories) was used as a tracer of intestinal perfusion(18).

Techniques: Expressing, Injection, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, SDS Page, Immunoprecipitation, Western Blot, Control

Journal: British Journal of Nutrition

Article Title: The human milk oligosaccharide 2′-fucosyllactose attenuates the severity of experimental necrotising enterocolitis by enhancing mesenteric perfusion in the neonatal intestine

doi: 10.1017/s0007114516002944

Figure Lengend Snippet: Fig. 6. The human milk oligosaccharide (HMO) 2′-fucosyllactose does not protect against necrotising enterocolitis (NEC) or enhance intestinal perfusion in endothelial nitric oxide synthase (eNOS)−/−mice. (a) Representative haematoxylin–eosin micrographs of the terminal ileum of eNOS−/−mice in the indicated treatment groups. (b) Representative confocal micrographs of terminal ileal whole mounts from eNOS−/−mice that received HMO and were either breast-fed (BF + HMO) or were formula-fed (FF + HMO). Whole mounts were stained for platelet endothelial cell adhesion molecule 1 (PECAM-1) to assess the microvasculature of the intestine and subjected to intra-cardiac injections with tomato lectin as a marker of intestinal perfusion (green). (c) Graph representing villus perfusion index as described in the Materials and methods section expressed as a percentage. (d) NEC severity score assessed by a pathologist blinded to the study conditions. (e) IL-1β mRNA expression from the terminal ileum of the mice in the indicated treatment groups. * P < 0·05 v. breast-fed control animals, ** P < 0·05 v. BF + HMO. Values are means with their standard errors. Scale bar is 50 μm. Representative of at least three separate experiments, where each mouse represents an individual symbol on a graph. qRT-PCR, quantitative real-time PCR.

Article Snippet: The fluorescein-labelled Lycopersicon esculentum (tomato) lectin (Vector Laboratories) was used as a tracer of intestinal perfusion(18).

Techniques: Staining, Marker, Expressing, Control, Quantitative RT-PCR, Real-time Polymerase Chain Reaction