Journal: American Journal of Physiology - Gastrointestinal and Liver Physiology

Article Title: Prematurity reduces citrulline-arginine-nitric oxide production and precedes the onset of necrotizing enterocolitis in piglets

doi: 10.1152/ajpgi.00198.2018

Figure Lengend Snippet: Multiple end-point measurements of necrotizing enterocolitis (NEC) incidence and severity in piglets. A: Kaplan-Meir graph detailing piglet survival. B: gross NEC score in piglets of advancing gestational age exposed to the NEC protocol. A threshold of 3 in any one segment was considered positive for NEC (dashed line). C: histological NEC score was determined in hematoxylin-eosin-stained tissue sections, and a threshold of ≥2 in any tissue section was considered positive for NEC (dashed line). D: plasma intestinal fatty-acid binding protein (iFABP) concentration in the final plasma sample for piglet groups in response to the NEC protocol. B–D: *P < 0.05, significant differences between groups, and error bars show 95% confidence interval. Data analyzed by Kruskal-Wallis test and Dunn’s multiple comparison test. NS, not significant.

Article Snippet: Amino acid concentrations in plasma and feed were measured using phenyl isothiocyanate derivatives relative to a methionine sulfone internal standard by HPLC ( 6 ). iFABP was measured using a human iFABP Quantikine ELISA Kit (R&D Systems).

Techniques: Staining, Clinical Proteomics, Binding Assay, Concentration Assay, Comparison

Journal: The Journal of international medical research

Article Title: I-FABP protein/mRNA and IL-6 as biomarkers of intestinal barrier dysfunction in neonates with necrotizing enterocolitis and SPF BALB/c mouse models.

doi: 10.1177/03000605241254788

Figure Lengend Snippet: Figure 2. Representative western blot bands of ileal tissue protein samples from the two neonatal mouse groups. Intestinal fatty acid binding protein (I-FABP) relative expression levels were significantly higher (P < 0.05) in tissues from the model group (1.273 0.376 pg/mL) than in those from the con- trol group (0.623 0.266 pg/mL).

Article Snippet: After blocking, the primary (b-actin, 1:1000; Abcam, Cambridge, United Kingdom and I-FABP, 1:2000; Proteintech Group, Inc., Wuhan, China) and secondary (mice antibody l: 5000, model HS201; TransGen Biotech, Inc., Beijing, China) antibodies were placed on the membrane and incubated at room temperature for 2 hours.

Techniques: Western Blot, Binding Assay, Expressing

Journal: The Journal of international medical research

Article Title: I-FABP protein/mRNA and IL-6 as biomarkers of intestinal barrier dysfunction in neonates with necrotizing enterocolitis and SPF BALB/c mouse models.

doi: 10.1177/03000605241254788

Figure Lengend Snippet: Figure 3. Comparison of intestinal fatty acid binding protein (I-FABP) translocation in ileal tissues from the two groups of neonatal mice. In the model group images, the yellow arrows point to I-FABP aggregation. Compared with the control group tissues, I-FABP expression was significantly increased in the model group tissues in the brush border membrane, central lacteal villi, basement membrane, and small intestine sub- mucosa. The images in panels a, b, and c are 20 magnification, with the scale bar representing 100 lM; the images in panels d, e, and f are 80 magnification, with the scale bar representing 25 lM. DAPI, 2-(4-Amidinophenyl)-6-indolecarbamidine dihydrochloride.

Article Snippet: After blocking, the primary (b-actin, 1:1000; Abcam, Cambridge, United Kingdom and I-FABP, 1:2000; Proteintech Group, Inc., Wuhan, China) and secondary (mice antibody l: 5000, model HS201; TransGen Biotech, Inc., Beijing, China) antibodies were placed on the membrane and incubated at room temperature for 2 hours.

Techniques: Comparison, Binding Assay, Translocation Assay, Control, Expressing, Membrane

Journal: The Journal of international medical research

Article Title: I-FABP protein/mRNA and IL-6 as biomarkers of intestinal barrier dysfunction in neonates with necrotizing enterocolitis and SPF BALB/c mouse models.

doi: 10.1177/03000605241254788

Figure Lengend Snippet: Figure 4. Serum intestinal fatty acid binding protein (I-FABP), I-FABP mRNA, and interleukin-6 (IL-6) levels in human newborn samples. Normality tests were performed first using GraphPad software. (a) The Shapiro–Wilk test results showed that neither data set was normally distributed. Nonparametric tests (Mann–Whitney U test) were therefore used for statistical analysis. The median value was 0.255 for the control group and 1.598 for the experimental group. We selected the mean value with the 95% confidence interval (CI) for plotting. I-FABP mRNA levels were significantly higher (P < 0.05) in the experimental group (5.104 7.333) than in the control group (3.556 8.939). (b) The Shapiro–Wilk test results showed that the control group data were normally distributed, while the experimental group data were not normally dis- tributed. Nonparametric tests (Mann–Whitney U test) were used for statistical analysis. The median value was 208.3 for the control group and 297.7 for the experimental group. I-FABP levels were significantly higher (P < 0.05) in the experimental group (370.306 264.621 pg/mL) than in the control group (232.083 210.669 pg/mL) and (c) the Shapiro–Wilk test results showed that neither data set was normally distributed. Nonparametric tests (Mann–Whitney U test) were used for statistical analysis. The median value was 1.5 for the control group and 26.3 for the experimental group. We selected the mean value with the 95% CI for plotting. IL-6 levels were significantly higher (P < 0.05) in the experimental group (319.277 971.174 pg/mL) than in the control group (15.327 28.355 pg/mL).

Article Snippet: After blocking, the primary (b-actin, 1:1000; Abcam, Cambridge, United Kingdom and I-FABP, 1:2000; Proteintech Group, Inc., Wuhan, China) and secondary (mice antibody l: 5000, model HS201; TransGen Biotech, Inc., Beijing, China) antibodies were placed on the membrane and incubated at room temperature for 2 hours.

Techniques: Binding Assay, Software, MANN-WHITNEY, Control

Journal: The Journal of international medical research

Article Title: I-FABP protein/mRNA and IL-6 as biomarkers of intestinal barrier dysfunction in neonates with necrotizing enterocolitis and SPF BALB/c mouse models.

doi: 10.1177/03000605241254788

Figure Lengend Snippet: Figure 5. Receiver operating characteristic (ROC) curve analysis of intestinal fatty acid binding protein (I-FABP), I-FABP mRNA, and interleukin-6 (IL-6) levels in human neonatal necrotizing enterocolitis (NEC) samples. The area under the ROC curve (AUC) value for the combined detection of I-FABP, I-FABP mRNA, and IL-6 was 0.768 (95% confidence interval: 0.650–0.885), with a sensitivity of 56.7%, specificity of 90%, negative prediction rate of 67.5%, and positive prediction rate of 85.0%.

Article Snippet: After blocking, the primary (b-actin, 1:1000; Abcam, Cambridge, United Kingdom and I-FABP, 1:2000; Proteintech Group, Inc., Wuhan, China) and secondary (mice antibody l: 5000, model HS201; TransGen Biotech, Inc., Beijing, China) antibodies were placed on the membrane and incubated at room temperature for 2 hours.

Techniques: Binding Assay

Journal: PLoS ONE

Article Title: Effects of Weaning on Intestinal Upper Villus Epithelial Cells of Piglets

doi: 10.1371/journal.pone.0150216

Figure Lengend Snippet: Effect of weaning on the expression of I-FABP, Bcl-2, and caspase-3 in jejunal upper villus epithelial cells of piglets.

Article Snippet: Antibodies for eIF4E, I-FABP, Bcl2, and Caspase3 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA), and antibodies for S6K, phospho-S6K (Thr389), mTOR, phospho-mTOR (Ser2448), 4EBP1, phospho-4EBP1 (Thr70) and β-actin were purchased from Cell Signaling Technology (Cedarlane, ON, Canada).

Techniques: Expressing

Journal: Microbial pathogenesis

Article Title: Outer membrane vesicles derived from heatstroke-associated intestinal microbiota promote multiple organ injury in mice.

doi: 10.1016/j.micpath.2022.105653

Figure Lengend Snippet: Fig. 1. Outer membrane vesicle (OMV) obtained from mice intestinal microbiota characterization. (a) OMV morphology monitored by transmission electron mi croscopy. The white arrow indicates the membrane bilayer surrounding the 150–200 nm vesicles. Bar = 100 nm. (b) OMV size distribution measured by nanoparticle tracking analysis (NTA). The diameter was roughly 150–200 nm. (c) Left panel: Typical western blot bands for OMVs as well as relevant proteins extracted from the supernatants, including OmpA. Right panel: Analysis of OmpA levels using the Gel-Pro image analyzer based on GADPH. (*) p < 0.05, (**) p < 0.01 compared with the control-supernatant, (##) p < 0.01 compared with the HS-supernatant by Kruskal-Wallis H test. (d) HS (heatstroke) and control OMV contents measured by NTA. (*) p < 0.05, (**) p < 0.01, (***) p < 0.001 by one-way ANOVA with a post hoc test. Each experiment was performed in triplicate.

Article Snippet: Enzyme-linked immunosorbent assay (ELISA) kits were used to measure the serum contents of IL-1β (Raybiotech, Atlanta, USA, ELMIL1b-1), IL-6 (Raybiotech ELM-IL6-1), intestinal mechanical barrier injury markers, such as ɑ-glutathione S-transferase (ɑ-GST, Solarbio Life Science, Beijing, China, BC0355), diamine oxidase (DAO, Bioss Antibodies, Woburn, MA, USA, AK066), and intestinal fatty acid binding protein (I-FABP, Novus Biologicals, Littleton, CO, USA, NBP2-82214), and the biological barrier injury markers D-lactate (Abcam, ab83429) and citrulline (Sigma-Aldrich, St. Louis, MO, USA, MAK423-1 KT) in accordance with the respective protocols.

Techniques: Membrane, Transmission Assay, Western Blot, Control

Journal: Microbial pathogenesis

Article Title: Outer membrane vesicles derived from heatstroke-associated intestinal microbiota promote multiple organ injury in mice.

doi: 10.1016/j.micpath.2022.105653

Figure Lengend Snippet: Fig. 7. OMVs derived from heatstroke mice (HS-OMVs) cause intestinal injury and inflammation in vivo. (a) H&E staining of the intestinal tissue. Scale bar = 100 μm. (b) Expression of ZO-1 in intestinal tissue by immunohistochemistry (IHC). (c) Expression of occludin in intestinal tissue by IHC. (d) Serum ɑ-GST, DAO, I-FABP, D- lactate, and citrulline levels in mice. (***) p < 0.001, (**) p < 0.01, (*) p < 0.05. Data are expressed as mean ± SEM. The error bars denote SEM.

Article Snippet: Enzyme-linked immunosorbent assay (ELISA) kits were used to measure the serum contents of IL-1β (Raybiotech, Atlanta, USA, ELMIL1b-1), IL-6 (Raybiotech ELM-IL6-1), intestinal mechanical barrier injury markers, such as ɑ-glutathione S-transferase (ɑ-GST, Solarbio Life Science, Beijing, China, BC0355), diamine oxidase (DAO, Bioss Antibodies, Woburn, MA, USA, AK066), and intestinal fatty acid binding protein (I-FABP, Novus Biologicals, Littleton, CO, USA, NBP2-82214), and the biological barrier injury markers D-lactate (Abcam, ab83429) and citrulline (Sigma-Aldrich, St. Louis, MO, USA, MAK423-1 KT) in accordance with the respective protocols.

Techniques: Derivative Assay, In Vivo, Staining, Expressing, Immunohistochemistry

Journal: Microbial pathogenesis

Article Title: Outer membrane vesicles derived from heatstroke-associated intestinal microbiota promote multiple organ injury in mice.

doi: 10.1016/j.micpath.2022.105653

Figure Lengend Snippet: Fig. 8. OMVs derived from heatstroke mice (HS-OMVs) cause intestinal inflammation in vivo. (a) MPO staining of intestinal tissues by immunohistochemistry (IHC). Scale bar = 100 μm. (b) F4/80 staining of intestinal tissues by IHC. Scale bar = 100 μm.

Article Snippet: Enzyme-linked immunosorbent assay (ELISA) kits were used to measure the serum contents of IL-1β (Raybiotech, Atlanta, USA, ELMIL1b-1), IL-6 (Raybiotech ELM-IL6-1), intestinal mechanical barrier injury markers, such as ɑ-glutathione S-transferase (ɑ-GST, Solarbio Life Science, Beijing, China, BC0355), diamine oxidase (DAO, Bioss Antibodies, Woburn, MA, USA, AK066), and intestinal fatty acid binding protein (I-FABP, Novus Biologicals, Littleton, CO, USA, NBP2-82214), and the biological barrier injury markers D-lactate (Abcam, ab83429) and citrulline (Sigma-Aldrich, St. Louis, MO, USA, MAK423-1 KT) in accordance with the respective protocols.

Techniques: Derivative Assay, In Vivo, Staining, Immunohistochemistry

Journal: Microbial pathogenesis

Article Title: Outer membrane vesicles derived from heatstroke-associated intestinal microbiota promote multiple organ injury in mice.

doi: 10.1016/j.micpath.2022.105653

Figure Lengend Snippet: Fig. 9. OMVs derived from heatstroke mice (HS-OMVs) cause intestine inflammation in vivo. (a) Expression of IL-1β in intestinal tissue by immunohistochemistry (IHC). (b) Expression of IL-6 in intestinal tissue by IHC. (c) Expression of TNF-ɑ in intestinal tissue by IHC.

Article Snippet: Enzyme-linked immunosorbent assay (ELISA) kits were used to measure the serum contents of IL-1β (Raybiotech, Atlanta, USA, ELMIL1b-1), IL-6 (Raybiotech ELM-IL6-1), intestinal mechanical barrier injury markers, such as ɑ-glutathione S-transferase (ɑ-GST, Solarbio Life Science, Beijing, China, BC0355), diamine oxidase (DAO, Bioss Antibodies, Woburn, MA, USA, AK066), and intestinal fatty acid binding protein (I-FABP, Novus Biologicals, Littleton, CO, USA, NBP2-82214), and the biological barrier injury markers D-lactate (Abcam, ab83429) and citrulline (Sigma-Aldrich, St. Louis, MO, USA, MAK423-1 KT) in accordance with the respective protocols.

Techniques: Derivative Assay, In Vivo, Expressing, Immunohistochemistry

Journal: EMBO Reports

Article Title: Terminal differentiation of villus tip enterocytes is governed by distinct Tgfβ superfamily members

doi: 10.15252/embr.202256454

Figure Lengend Snippet:

Article Snippet: Fabp2 (goat, polyclonal) , R&DSystems , AF1486.

Techniques: Generated, Recombinant, Expressing, Plasmid Preparation, Transduction, Sequencing, shRNA, Gentle, Software, Positive Control, Negative Control, SYBR Green Assay, Activity Assay