Journal: Nutrients

Article Title: Effects of Different Routes and Forms of Vitamin D Administration on Mesenteric Lymph Node CD4+ T Cell Polarization and Intestinal Injury in Obese Mice Complicated with Polymicrobial Sepsis.

doi: 10.3390/nu14173557

Figure Lengend Snippet: Figure 5. Cathelicidin concentrations in ileum tissues after cecal ligation and puncture at two time points. Data are presented as the mean ± standard error of the mean (SEM). n = 8 for each group. The groups are described in the legend of Figure 1. Differences among groups at the same time point were analyzed by a one-way analysis of variance using Tukey’s post hoc test. * Significantly differs from the other groups at the same time point (p < 0.05).

Article Snippet: After centrifugation, supernatants were used to quantify cathelicidin levels with a Mouse Cathelicidin Antimicrobial Peptide (CAMP) ELISA kit (ABclonal, Woburn, MA, USA).

Techniques: Ligation

Journal: PLoS ONE

Article Title: Evidence Suggesting a Role of Iron in a Mouse Model of Nephrogenic Systemic Fibrosis

doi: 10.1371/journal.pone.0136563

Figure Lengend Snippet: Primary antibodies used in this study. IHC—immunohistochemistry, WB—western blot.

Article Snippet: Anti-hepcidin , Goat (polyclonal) , 1:1,000 WB , Novus Biologicals , NBP1-59337.

Techniques:

Journal: bioRxiv

Article Title: Live-cell invasive phenotyping uncovers the ALK2/BMP6 iron homeostasis pathway as a therapeutic vulnerability in LKB1-mutant lung cancer

doi: 10.1101/2023.06.14.544941

Figure Lengend Snippet: A) Schematic depicting pre-clinical trial workflow. B) Brightfield images of tumors isolated from vehicle and LDN214117 treated mice. C) Mean tumor volume from mice treated with vehicle ( 6 mice/group) and LDN214117 (7 mice/group). D) Graph depicting mean tumor weight from vehicle and LDN214117 treated mice. E) Western blot to assess BMP6 and hepcidin expression level in tumors from vehicle and LDN214117-treated mice. F) Representative images of immunohistochemistry for BMP6 and Hepcidin on tumor sections from mice treated with vehicle or LDN214117. F) Model depicting mechanism of altered iron homeostasis in LKB1-mutant tumor cells.

Article Snippet: Hepcidin Antimicrobial Peptide antibody (NBP1-59337) was from Novus Biologicals.

Techniques: Isolation, Western Blot, Expressing, Immunohistochemistry, Mutagenesis

Journal: Frontiers in Endocrinology

Article Title: Elevated blood levels of liver-expressed antimicrobial peptide 2 in patients with insulinoma and its expression in insulinomas

doi: 10.3389/fendo.2025.1685806

Figure Lengend Snippet: Fasting serum levels of LEAP2 in patients and control group and the relationship between serum LEAP2 levels and obesity. (A) Serum levels of LEAP2 in patients with insulinoma were significantly higher than those in control groups (P = 0.028). (B) Serum LEAP2 levels in obese patients were not significantly higher than those in non-obese patients (P= 0.10). (C) Serum levels of LEAP2 in patients showed a trend toward correlation with obesity (P = 0.067). (D, E) Serum levels of LEAP2 in control group significantly correlated with obesity. Each dot represents the LEAP2 level of an individual patient or control.

Article Snippet: For immunofluorescence staining and co-localization, selected specimens were stained with anther LEAP2 antibody (Abbexa, abx104118, Cambridge, UK; dilution 1:25) and, where indicated, co-stained with an insulin antibody (Santa Cruz Biotechnology, 2D11-H5, sc-8033, Lot #G2904; dilution 1:100).

Techniques: Control

Journal: Frontiers in Endocrinology

Article Title: Elevated blood levels of liver-expressed antimicrobial peptide 2 in patients with insulinoma and its expression in insulinomas

doi: 10.3389/fendo.2025.1685806

Figure Lengend Snippet: Relationship between fasting serum levels of LEAP2 and insulin. (A) LEAP2 levels in patients with hyperinsulinemia (> 17.2 µIU/ml) were significantly higher than those in patients with normal insulin levels (P = 0.021). (B) Serum LEAP2 levels positively correlated with insulin levels in patients with insulinoma. (C) Serum levels of LEAP2 showed a trend toward correlation with insulin levels but did not reach significance (P=0.06) in controls.

Article Snippet: For immunofluorescence staining and co-localization, selected specimens were stained with anther LEAP2 antibody (Abbexa, abx104118, Cambridge, UK; dilution 1:25) and, where indicated, co-stained with an insulin antibody (Santa Cruz Biotechnology, 2D11-H5, sc-8033, Lot #G2904; dilution 1:100).

Techniques:

Journal: Frontiers in Endocrinology

Article Title: Elevated blood levels of liver-expressed antimicrobial peptide 2 in patients with insulinoma and its expression in insulinomas

doi: 10.3389/fendo.2025.1685806

Figure Lengend Snippet: Immunohistochemical staining of LEAP2 and immunofluorescence co-staining of LEAP2 and insulin in insulinoma. (A) IHC staining of LEAP2 in an insulinoma, relatively strong expression of LEAP2 in tumor (left part) and no expression of LEAP2 in para-tumoral pancreatic tissue (right part). (B) Relatively weak expression of LEAP2 in anther insulinoma (left part) and no expression of LEAP2 in para-tumoral pancreatic tissue (right part). (C) Insulin staining (red) using an anti-insulin antibody (Santa Cruz Biotechnology, 2D11-H5, sc-8033; dilution 1:100). (D) LEAP2 staining (green) using an anti-LEAP2 antibody (Abbexa, abx104118; dilution 1:25). (E) Merged image showing co-localization of LEAP2 with insulin in tumor cells. Nuclei were counterstained with DAPI (blue).

Article Snippet: For immunofluorescence staining and co-localization, selected specimens were stained with anther LEAP2 antibody (Abbexa, abx104118, Cambridge, UK; dilution 1:25) and, where indicated, co-stained with an insulin antibody (Santa Cruz Biotechnology, 2D11-H5, sc-8033, Lot #G2904; dilution 1:100).

Techniques: Immunohistochemical staining, Staining, Immunofluorescence, Immunohistochemistry, Expressing

Journal: Frontiers in Endocrinology

Article Title: Elevated blood levels of liver-expressed antimicrobial peptide 2 in patients with insulinoma and its expression in insulinomas

doi: 10.3389/fendo.2025.1685806

Figure Lengend Snippet: Schematic illustration of the potential interplay between LEAP2, ghrelin, insulin, and obesity. Chronic hyperinsulinemia may contribute to increased circulating LEAP2 and suppressed ghrelin levels, which could together reduce appetite and food intake. This proposed feedback model summarizes possible mechanisms supported by our findings.

Article Snippet: For immunofluorescence staining and co-localization, selected specimens were stained with anther LEAP2 antibody (Abbexa, abx104118, Cambridge, UK; dilution 1:25) and, where indicated, co-stained with an insulin antibody (Santa Cruz Biotechnology, 2D11-H5, sc-8033, Lot #G2904; dilution 1:100).

Techniques:

Journal: Gut Microbes

Article Title: Underlying MASLD-induced gut microbiome dysbiosis and intestinal inflammation are key to poor outcomes in vibriosis infections in a preclinical model

doi: 10.1080/19490976.2026.2652474

Figure Lengend Snippet: Establishment of MASLD pathophysiology in mice by CD-HFD feeding for 20 weeks. (A) Schematic representation of the experimental mouse groups: LEAN [ n = 6; chow diet-fed mice for 20 weeks] and MASLD [ n = 6; CD-HFD fed mice for 20 weeks]. (B) Liver weight (grams), and (C) liver-to-body weight ratio of the LEAN and MASLD mouse groups. Serum levels of (D) ALT (U/L), (E) AST (U/L), (F) hepcidin (ng/mL), and (G) ferritin (μg/mL) in the LEAN and MASLD groups. Formalin-fixed, paraffin-embedded 5 μm liver slices from the LEAN and MASLD groups were used for histopathological analyses. Representative images of (H) hematoxylin and eosin (H&E) staining, picrosirius red (PSR) staining, and immunohistochemistry images depicting α -SMA, and IL-1β immunoreactivity (indicated by black arrowheads) in the liver sections of LEAN and MASLD mouse groups. H&E and immunohistochemistry images were captured at 200× magnification, whereas PSR images were captured at 100× magnification. (I) NAFLD activity score (NAS) and fibrosis score for the LEAN and MASLD groups. Morphometric analyses (calculated as %ROI) of (J) PSR staining, (K) α -SMA, and (L) IL-1β immunoreactivity, where the Y-axis represents % positive immunoreactive area ( n = 3; mean value taken from three separate microscopic fields). The data are presented as mean ± SEM, and statistical significance was tested using unpaired t-test between the two groups, followed by Bonferroni post-hoc corrections (* p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Serum hepcidin (ng/mL) (Catalog No. NBP2-82129, Novus Biologicals, Centennial, CO, USA), ferritin (μg/mL) (Catalog No. KA1941, Novus Biologicals, Centennial, CO, USA), immunoglobulin-A (IgA) (μg/mL) (Catalog No. ab157717, Abcam, Cambridge, MA, USA) and C-reactive protein (CRP) (μg/mL) (Catalog No. MCRP00, Novus Biological, Centennial, CO, USA) were quantified as per the manufacturers’ instructions using commercially available ELISA kits.

Techniques: Formalin-fixed Paraffin-Embedded, Staining, Immunohistochemistry, Activity Assay

Journal: Gut Microbes

Article Title: Underlying MASLD-induced gut microbiome dysbiosis and intestinal inflammation are key to poor outcomes in vibriosis infections in a preclinical model

doi: 10.1080/19490976.2026.2652474

Figure Lengend Snippet: Underlying MASLD conditions in mice caused increased non-cholera vibriosis and affected the hepatic pathophysiology. (A) Schematic representation of the experimental mouse groups: LEAN [ n = 6; chow diet-fed mice for 20 weeks], LEAN + VV [ n = 6; mice fed with chow diet for 20 weeks and received oral VV inoculation for 24 h], MASLD [ n = 6; CD-HFD fed mice for 20 weeks], and MASLD + VV [ n = 6; mice fed with CD-HFD for 20 weeks and received oral VV inoculation for 24 h]. (B) Liver weight (grams), and (C) liver-to-body weight ratio of the LEAN, LEAN + VV, MASLD, and MASLD + VV mouse groups. Serum levels of (D) CRP (μg/mL), (E) ALT (U/L), (F) AST (U/L), (G) hepcidin (ng/mL), and (H) ferritin (μg/mL) in the LEAN, LEAN + VV, MASLD, and MASLD + VV mouse groups. Formalin-fixed, paraffin-embedded 5 μm liver slices from the LEAN, LEAN + VV, MASLD, and MASLD + VV mouse groups were used for histopathological analyses. Representative images of (I) hematoxylin and eosin (H&E) staining and Picrosirius red (PSR) staining of liver sections of LEAN, LEAN + VV, MASLD, and MASLD + VV mouse groups. H&E images were captured at 200× magnification, whereas PSR images were captured at 100× magnification. (J) NAFLD activity score (NAS) and fibrosis score for the LEAN, LEAN + VV, MASLD, and MASLD + VV mouse groups. (K) Morphometric analyses (calculated as %ROI) of PSR staining, where the Y-axis represents % positive immunoreactive area ( n = 3; mean value taken from three separate microscopic fields). Data were represented as mean ± SEM, and statistical significance was tested using one-way ANOVA between all the groups, followed by Bonferroni post-hoc corrections (ns = non-significant, * p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Serum hepcidin (ng/mL) (Catalog No. NBP2-82129, Novus Biologicals, Centennial, CO, USA), ferritin (μg/mL) (Catalog No. KA1941, Novus Biologicals, Centennial, CO, USA), immunoglobulin-A (IgA) (μg/mL) (Catalog No. ab157717, Abcam, Cambridge, MA, USA) and C-reactive protein (CRP) (μg/mL) (Catalog No. MCRP00, Novus Biological, Centennial, CO, USA) were quantified as per the manufacturers’ instructions using commercially available ELISA kits.

Techniques: Formalin-fixed Paraffin-Embedded, Staining, Activity Assay

Journal: Gut Microbes

Article Title: Underlying MASLD-induced gut microbiome dysbiosis and intestinal inflammation are key to poor outcomes in vibriosis infections in a preclinical model

doi: 10.1080/19490976.2026.2652474

Figure Lengend Snippet: FMT in mice with underlying MASLD conditions showed improved pathophysiological outcomes Compared to non-cholera vibriosis infection. (A) Schematic representation of the experimental mouse groups: LEAN + VV [ n = 6; mice fed with chow diet for 20 weeks and received oral VV inoculation for 24 h], LEAN + FMT + VV [ n = 6; chow diet-fed mice, first treated with the ABX cocktail for 15 d followed by FMT for 7 d, and received oral VV inoculation for 24 h], MASLD + VV [ n = 6; mice fed with CD-HFD for 20 weeks and received oral VV inoculation for 24 h], and MASLD + FMT + VV [ n = 6; CD-HFD fed mice, first treated with the ABX cocktail for 15 d followed by FMT for 7 d, and received oral VV inoculation for 24 h]. (B) Liver weight (grams), and (C) liver-to-body weight ratio of the LEAN + VV, LEAN + FMT + VV, MASLD + VV, and MASLD + FMT + VV mouse groups. Serum levels of (D) CRP (μg/mL), (E) ALT (U/L), (F) AST (U/L), (G) Hepcidin (ng/mL), (H) Ferritin (μg/mL) in the LEAN + VV, LEAN + FMT + VV, MASLD + VV, and MASLD + FMT + VV mouse groups. Formalin-fixed, paraffin-embedded 5 μm liver slices were used for histopathological analyses. Representative images of (I) hematoxylin and eosin (H&E) staining and picrosirius red (PSR) staining in the liver sections from the LEAN + VV, LEAN + FMT + VV, MASLD + VV, and MASLD + FMT + VV mouse groups. H&E images were captured at 200× magnification, whereas PSR images were captured at 100× magnification. (J) NAFLD activity score (NAS) and fibrosis score for the LEAN + VV, LEAN + FMT + VV, MASLD + VV, and MASLD + FMT + VV mouse groups. (K) Morphometric analyses (calculated as %ROI) of PSR staining, where the Y-axis represents % positive immunoreactive area ( n = 3; mean value taken from three separate microscopic fields). The data are presented as mean ± SEM, and statistical significance was tested using one-way ANOVA between all the groups, followed by Bonferroni post-hoc corrections (ns = non-significant, * p < 0.05, ** p < 0.01, *** p < 0.001).

Article Snippet: Serum hepcidin (ng/mL) (Catalog No. NBP2-82129, Novus Biologicals, Centennial, CO, USA), ferritin (μg/mL) (Catalog No. KA1941, Novus Biologicals, Centennial, CO, USA), immunoglobulin-A (IgA) (μg/mL) (Catalog No. ab157717, Abcam, Cambridge, MA, USA) and C-reactive protein (CRP) (μg/mL) (Catalog No. MCRP00, Novus Biological, Centennial, CO, USA) were quantified as per the manufacturers’ instructions using commercially available ELISA kits.

Techniques: Infection, Formalin-fixed Paraffin-Embedded, Staining, Activity Assay

Journal: Chinese Medical Journal

Article Title: LEAP2 ameliorates high fat diet-impaired glucose tolerance and enhances glucose-stimulated insulin secretion from β cells through PPARγ signal pathway

doi: 10.1097/cm9.0000000000003215

Figure Lengend Snippet: Figure 1: LEAP2 improved the glucose tolerance and pancreatic islet morphology in vivo and promoted GSIS in vitro. (A) Experimental protocol for constructing the mouse T2D model and LEAP2 treatment. (B) Determination of rate of body weight change after LEAP2 treatment in T2D and control mice (n = 7). (C,D) Measurement of blood glucose during IGTT after LEAP2 treatment in T2D and control mice (n = 5–7). (E) Representative hematoxylin and eosin staining images of pancreases after LEAP2 treatment in T2D and control mice (n = 5). From left to right: Control group, standard chow diet mice were intraperitoneally injected with saline; LEAP2 group, standard chow diet mice were intraperitoneally injected with LEAP2 (30 µg/kg); T2D group, T2D mice were intraperitoneally injected with saline; T2D+LEAP2 group, T2D mice were intraperitoneally injection with LEAP2 (30 µg/kg). All treatments were carried out at the same time in the experiment, after 14 days of continuous treatment with LEAP2 or saline. Scale bars = 50 µm. (F) LEAP2 significantly promoted MIN6 cells insulin secretion in the conditions of 3 mmol/L (Control and LEAP2 group) or 18 mmol/L (HG and HG + LEAP2 group) glucose (n = 3). (G) Measurement of intracellular Gck levels treated by LEAP2 in MIN6 cells (n = 6). (H) Measurement of intracellular ATP level treated by LEAP2 in MIN6 cells (n = 6). (I) Measurement of intracellular Ca2+ level treated by LEAP2 in MIN6 cells (n = 6). (J,K) PPARγ level in MIN6 cells treated with LEAP2 by Western blotting (n = 3). (L) Measurement of insulin content in MIN6 cells treated with LEAP2 or GW9662 (n = 3). (M) Measurement of Gck content in MIN6 cells treated with LEAP2 or GW9662 (n = 3). (N) GHSR siRNA significantly downregulated GHSR expression in mRNA levels (n = 3). (O) Measurement of insulin content in MIN6 cells treated with LEAP2 after knockdown of GHSR (n = 3). *P <0.05, **P <0.01, ***P <0.001, ****P <0.0001. All data are presented as the mean ± SD. AUC: Area under the curve; HFD: High-fat diet; HG: High glucose; GHSR: Growth hormone secretagogue receptor; IGTT: Intraperitoneal glucose tolerance test; LEAP2: Liver-expressed antimicrobial peptide 2; ns: Not significant; prot: Protein; PPAR: Poly (ADP-ribose) polymerase; siRNA: small interfering RNA; SD: Standard deviation; STZ: Streptozotocin; T2D: Type 2 diabetes.

Article Snippet: However, LEAP2 treatment had no effects on the protein expression of Akt and phosphorylated Akt in MIN6 cells [Supplementary Figure 2K,L, http://links.lww.com/CM9/C78] To investigate the activation of PPARγ in the secretory effects of LEAP2 on insulin secretion, MIN6 cells were incubated with the PPARγ-specific antagonist, GW9662 (catalog No. HY-16578, MedChemExpress, Shanghai, China), for 12 hours in the presence and absence of LEAP2.

Techniques: In Vivo, In Vitro, Control, Staining, Injection, Saline, Western Blot, Expressing, Knockdown, Small Interfering RNA, Standard Deviation

Journal: bioRxiv

Article Title: Synthetic Mucus Biomaterials for Antimicrobial Peptide Delivery

doi: 10.1101/2023.03.07.531025

Figure Lengend Snippet: Physical characterization of LL37-SM hydrogels. ( A ) Schematic of LL37-SM hydrogel preparation. ( B ) Degradation profile of LL37-SM hydrogels over 24 hours ( n = 3). ( C ) Swelling ratio of LL37-SM hydrogels after 4 hours in PBS ( n = 3). No statistical significance observed by one-way ANOVA.

Article Snippet: LL37 loaded SM hydrogels were made in cylindrical molds to create 100 μL hydrogel discs with varying concentrations of fluorescently labelled tetramethylrhodamine-LL37 (TRITC-LL37, Rockland).

Techniques:

Journal: bioRxiv

Article Title: Synthetic Mucus Biomaterials for Antimicrobial Peptide Delivery

doi: 10.1101/2023.03.07.531025

Figure Lengend Snippet: Microrheological characterization of LL37-SM hydrogels. ( A ) Measured l o g 10 ⁡ M S D 1 s of 100 n m PEG-coated NP in LL37-SM hydrogels after 24 hours of incubation at room temperature ( n = 20 ) . * * * P < 0.001 and * * * * P < 0.0001 for Kruskal-Wallis test. ( B ) Elastic and viscous moduli ratio G ′ / G ″ at a frequency ω = 1 H z calculated from measured M S D ⁡ ( n = 6 ) . No statistical significance observed by Kruskal-Wallis test. ( C ) Mean complex microviscosity η * at a frequency ω = 1 H z calculated from measured MSD ( n = 3 ) . * P < 0.05 for one-way ANOVA. The color of the asterisk indicates the comparison group.

Article Snippet: LL37 loaded SM hydrogels were made in cylindrical molds to create 100 μL hydrogel discs with varying concentrations of fluorescently labelled tetramethylrhodamine-LL37 (TRITC-LL37, Rockland).

Techniques: Incubation, Comparison

Journal: bioRxiv

Article Title: Synthetic Mucus Biomaterials for Antimicrobial Peptide Delivery

doi: 10.1101/2023.03.07.531025

Figure Lengend Snippet: Release kinetics of LL37 from SM hydrogels. ( A ) FRAP mean relative fluorescence intensity profile of LL37-SM hydrogels ( n = 3). ( B ) Schematic of TRITC-LL37-SM hydrogel set up for evaluating release. Cumulative mass release profiles of LL37 from SM hydrogels ( n = 3) ( C ) without and ( D ) with 200 ng/mL trypsin over 24 hours ( n = 3).

Article Snippet: LL37 loaded SM hydrogels were made in cylindrical molds to create 100 μL hydrogel discs with varying concentrations of fluorescently labelled tetramethylrhodamine-LL37 (TRITC-LL37, Rockland).

Techniques: Fluorescence

Journal: bioRxiv

Article Title: Synthetic Mucus Biomaterials for Antimicrobial Peptide Delivery

doi: 10.1101/2023.03.07.531025

Figure Lengend Snippet: Release profiles were fitted to the Korsmeyer-Peppas model.

Article Snippet: LL37 loaded SM hydrogels were made in cylindrical molds to create 100 μL hydrogel discs with varying concentrations of fluorescently labelled tetramethylrhodamine-LL37 (TRITC-LL37, Rockland).

Techniques: Diffusion-based Assay

Journal: bioRxiv

Article Title: Synthetic Mucus Biomaterials for Antimicrobial Peptide Delivery

doi: 10.1101/2023.03.07.531025

Figure Lengend Snippet: Antimicrobial activity of LL37-SM hydrogels against planktonic PAO1. ( A ) Schematic of hydrogel treatment on planktonic GFP-PAO1 and PAO1 to assess the effect on growth and viability, respectively. ( B ) Growth profiles of planktonic PAO1 with treatment ( n = 3). ( C ) PAO1 viability after 3 hours of treatment ( n = 3). **** P <0.0001 for two-way ANOVA. Blue asterisks indicate the comparison between time groups.

Article Snippet: LL37 loaded SM hydrogels were made in cylindrical molds to create 100 μL hydrogel discs with varying concentrations of fluorescently labelled tetramethylrhodamine-LL37 (TRITC-LL37, Rockland).

Techniques: Activity Assay, Comparison