Journal: Cell Reports Medicine

Article Title: LEAP2 reduces postprandial glucose excursions and ad libitum food intake in healthy men

doi: 10.1016/j.xcrm.2022.100582

Figure Lengend Snippet: Overview of the clinical study design The study was designed as a randomized, double-blind, placebo-controlled, crossover study including 20 healthy, young men. IC, indirect calorimetry; LEAP2, liver-expressed antimicrobial peptide 2; TC, thermal imaging camera; VAS, visual analog scale.

Article Snippet: LEAP2 peptide (human studies) , Vivitide (previously named Peptides International) , N/A.

Techniques: Imaging

Journal: Cell Reports Medicine

Article Title: LEAP2 reduces postprandial glucose excursions and ad libitum food intake in healthy men

doi: 10.1016/j.xcrm.2022.100582

Figure Lengend Snippet: LEAP2 alters postprandial plasma concentrations of glucose, glucagon, and growth hormone during a liquid mixed meal test and fasting plasma concentrations of insulin, C-peptide, C-peptide/glucose ratio, glucagon, and glycerol in healthy, young men Plasma concentrations of LEAP2 (A), glucose (B), insulin (C), C-peptide (D), C-peptide/glucose ratio (E), glucagon (F), free fatty acids (G), glycerol (H), triglycerides (I), acetaminophen (J), growth hormone (K), and acyl ghrelin (L). Bold dotted line, infusion start (0 min); thin dotted line, liquid mixed meal test (65 min); blue square symbols, LEAP2 infusion; gray round symbols, placebo infusion (n = 20). Data are presented as mean ± SEM. Student’s paired t test of AUC 60–255 min : p = 0.017 (B), p < 0.001 (F), and p < 0.001 (K). Student’s paired t test of AUC 0–60 min : p < 0.001 (C), p = 0.018 (D), p = 0.003 (E), p = 0.038 (F), and p = 0.039 (H). AUC, area under the curve.

Article Snippet: LEAP2 peptide (human studies) , Vivitide (previously named Peptides International) , N/A.

Techniques: Clinical Proteomics

Journal: Cell Reports Medicine

Article Title: LEAP2 reduces postprandial glucose excursions and ad libitum food intake in healthy men

doi: 10.1016/j.xcrm.2022.100582

Figure Lengend Snippet: No effect on ratings of hunger and other sensations, resting energy expenditure, or brown adipose tissue thermogenesis during LEAP2 infusion in healthy, young men (A–G) Visual analog scale ratings of hunger (A), satiety (B), prospective food consumption (C), fullness (D), nausea (E), comfort (F), and thirst (G). (H) Resting energy expenditure. (I) Manubrium-corrected supraclavicular skin temperature. Bold dotted line, infusion start (0 min); thin dotted line, liquid mixed meal test (65 min); blue square symbols, LEAP2 infusion; gray round symbols, placebo infusion (n = 20). Data are presented as mean ± SEM. No significant differences (Student’s paired t test of area under the curve, [A–G]; repeated-measures mixed effects model, [H and I]).

Article Snippet: LEAP2 peptide (human studies) , Vivitide (previously named Peptides International) , N/A.

Techniques:

Journal: Cell Reports Medicine

Article Title: LEAP2 reduces postprandial glucose excursions and ad libitum food intake in healthy men

doi: 10.1016/j.xcrm.2022.100582

Figure Lengend Snippet: LEAP2 reduces food intake during an ad libitum meal test in healthy, young men Ad libitum food intake assessed by total intake (A), intake per kg body weight (B), and duration of the ad libitum meal (C). Blue square symbols, LEAP2 infusion; gray round symbols, placebo infusion (n = 20). Data are presented as mean ± SEM. ∗p < 0.05; ∗∗p < 0.01 (Student’s paired t test). Mean differences 432 (95% confidence interval [CI] 155–709) kJ, p = 0.0041 (A); 5.4 (95% CI 2.1–8.7) kJ/kg, p = 0.0029 (B); 1.5 (95% CI 0.2–2.9) min, p = 0.0303 (C).

Article Snippet: LEAP2 peptide (human studies) , Vivitide (previously named Peptides International) , N/A.

Techniques:

Journal: Cell Reports Medicine

Article Title: LEAP2 reduces postprandial glucose excursions and ad libitum food intake in healthy men

doi: 10.1016/j.xcrm.2022.100582

Figure Lengend Snippet: LEAP2 reduces postprandial plasma glucose and food intake in mice in a GHSR-dependent manner (A and B) Plasma glucose concentrations in wild-type (mean body weight = 22.3 g) and GHSR-null (GHSR −/− ) mice (mean body weight = 22.3 g) at baseline and 1, 2, and 4 h after LEAP2 or vehicle dosing (A) and cumulative food intake 1, 2, and 4 h after dosing (B). (C and D) Fasting insulin (C) and plasma glucose (D) concentrations in wild-type mice (mean body weight = 22.5 g). Blue square symbols, LEAP2 dosing; gray round symbols, vehicle dosing (n = 8–10). Data are presented as mean ± SEM. Repeated-measures two-way ANOVA: p = 0.0007 (B). ∗p < 0.05; ∗∗p < 0.01; ∗∗∗p < 0.001 (Šidák’s post hoc test). GHSR, growth hormone secretagogue receptor.

Article Snippet: LEAP2 peptide (human studies) , Vivitide (previously named Peptides International) , N/A.

Techniques: Clinical Proteomics

Journal: Cell Reports Medicine

Article Title: LEAP2 reduces postprandial glucose excursions and ad libitum food intake in healthy men

doi: 10.1016/j.xcrm.2022.100582

Figure Lengend Snippet:

Article Snippet: LEAP2 peptide (human studies) , Vivitide (previously named Peptides International) , N/A.

Techniques: Recombinant, Saline, RIA Assay, Software, Imaging

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Intracellular iron deficiency in pulmonary arterial smooth muscle cells induces pulmonary arterial hypertension in mice.

doi: 10.1073/pnas.1822010116

Figure Lengend Snippet: Fig. 1. Loss of HAMP responsiveness in PASMCs leads to PAH and right heart failure. (A) Representative images of α-SMA and FPN immunostaining in the lung 6 wk posttamoxifen induction (n = 6 mice per group). Original magnification 40×. (Scale bar: 25 μm.) (B) Representative images of α-SMA (red) and FPN (green) immunostaining in primary mouse PASMCs. Cells were treated with either vehicle or exogenous HAMP peptide (+HAMP). Original magnification 20×. (Scale bar: 100 μm.) (n = 3 mice) (C) Membrane FPN levels measured by flow cytometry in primary mouse PASMCs (n = 3 mice). (D) Representative images of α-SMA immunostaining in the lungs at 6 and 12 wk posttamoxifen induction (n = 6 per group). Original magnification 40×. (Scale bar 25: μm.) (E) Percentage of nonmuscularized (N), partially muscularized (P), and fully muscularized (F) vessels in lungs at 6 wk and 12 wk posttamoxifen induction (n = 5–6 per group). P values shown relative to the respective vessel category in control mice of the respective time point. (F–H) Measurements of right ventricular systolic pressure (RVSP) (n = 5–6), right ventricular ejection fraction (RVEF) (n = 5–12) and right ventricular end systolic volume (RVESV) (n = 5–12). (I) Representative mid- ventricular magnetic resonance images of the heart showing septal bowing at 6 wk post tamoxifen induction (n = 5–12 per group). (J) Ratio of RV to LV+Septum weight (n = 5–6 per group). (K) Representative images of WGA staining in RV (n = 5–6 per group). Original magnification 10×. (Scale bar: 200 μm.) Quantitation of cardiomyocyte area shown in Right. P values for paired comparisons were calculated using Student’s t test. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Mouse PASMCs were treated with mouse HAMP peptide (PLP-4434-s, Peptides International) while human PASMCs were treated with human HAMP peptide (PLP-4392-s, Peptides International) at a final concentration of 0.5 μmol/L for 12–16 h. Ferric citrate (F3388, Sigma-Aldrich) was dissolved in cell growth medium at a final concentration of 200 μmol/L.

Techniques: Immunostaining, Membrane, Flow Cytometry, Control, Staining, Quantitation Assay

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Intracellular iron deficiency in pulmonary arterial smooth muscle cells induces pulmonary arterial hypertension in mice.

doi: 10.1073/pnas.1822010116

Figure Lengend Snippet: Fig. 4. The HAMP/FPN axis operates cell autonomously in PASMCs and is dysregulated by BMPR2 mutations. (A) Representative images of FPN (green) and α-SMA (red) staining (Left) and FPN flow cytometry (Right) in mouse PASMCs. Original magnification 20×. (Scale bar: 100 um.) (B) Representative images of FPN (green) and α-SMA (red) staining (Left) and FPN flow cytometry (Right) in human PASMCs. Cells treated with vehicle or exogenous HAMP peptide (+HAMP) are also shown. Original magnification 20×. (Scale bar: 100 um.) (C–G) Levels of total elemental iron, cellular ferritin, and relative levels of tfr1, dmt1, and et-1 in mouse and human PASMCs treated with scrambled siRNA, hamp siRNA, vehicle, or HAMP peptide (+HAMP). (H) Relative levels of hamp mRNA expression in mouse and human PASMCs with intact BMPR2 or with the R899X mutation. (I) Representative images of FPN flow cytometry in mouse and human PASMCs. R899X cells were treated with vehicle (R899X) or exogenous HAMP peptide (R899X+HAMP). (J) ET-1 levels in supernatants from corresponding samples. (K) Fold induction by BMP6 of hamp mRNA in human and mouse PASMCs with intact BMPR2 or with the R899X mutation. P values are shown relative to R899X cells for the respective BMP6 concentration. (L) ET-1 levels in the supernatants of primary PASMCs derived from three healthy controls (HC1, -2, and -3) and three PAH patients with known BMPR2 mutations (PAH1, -2, and -3). Cells were treated with either vehicle, FAC, or HAMP peptide. P values shown are relative to vehicle-treated cells. (Right) Mean values for all HC vs. all PAH. (E–J and K) P values for paired comparisons were calculated using Student’s t test. (J and L) P values were calculated using ANOVA. Post hoc tests used Bonferroni correction to allow for multiple comparisons. *P < 0.05, **P < 0.01, ***P < 0.001. n = 3 biological replicates.

Article Snippet: Mouse PASMCs were treated with mouse HAMP peptide (PLP-4434-s, Peptides International) while human PASMCs were treated with human HAMP peptide (PLP-4392-s, Peptides International) at a final concentration of 0.5 μmol/L for 12–16 h. Ferric citrate (F3388, Sigma-Aldrich) was dissolved in cell growth medium at a final concentration of 200 μmol/L.

Techniques: Staining, Flow Cytometry, Expressing, Mutagenesis, Concentration Assay, Derivative Assay

Journal: Journal of Clinical and Translational Hepatology

Article Title: Preclinical Long-term Magnetic Resonance Imaging Study of Silymarin Liver-protective Effects

doi: 10.14218/JCTH.2021.00499

Figure Lengend Snippet: (A) Prussian blue staining evaluated iron accumulation. (B) Iron accumulation significantly decreased in the TAA-SY group compared with the TAA group. (C) Western blot assays of ferritin and hepcidin protein expression. (D) Ferritin in the TAA-SY group was decreased compared with the TAA group. Hepcidin in the TAA-SY group was slightly increased compared with the TAA group. (E) T 2 * values and iron overload (%) were negatively correlated (* p <0.05, ** p <0.01, *** p <0.001). Con, control; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; TAA, thioacetamide-injected; TAA-SY, TAA-injected and silymarin-treated.

Article Snippet: Western blots reacted with primary antibodies against ferritin (3998S, Cell Signaling Technology, Beverly, MA, USA), hepcidin (M01347, Boster Biological Technology, Pleasanton, CA, USA) and GAPDH (ab8245, Abcam, USA), followed by HRP-conjugated secondary antibodies.

Techniques: Staining, Western Blot, Expressing, Control, Injection