Journal: Journal of molecular and cellular cardiology

Article Title: Stem Cell Factor is Responsible for the Rapid Response in Mature Mast Cell Density in the Acutely Stressed Heart

doi: 10.1016/j.yjmcc.2012.07.011

Figure Lengend Snippet: Stem cell factor (SCF) concentration in left ventricular (LV) tissue slices (3A) and the conditioned medium (3B) and mature mast cell (MC) density (3C) that were incubated with media alone (control), compound 48/80 (20 μg/ml), or 48/80 plus chymostatin for 16 hrs. SCF levels in LV tissue slices and culture media were normalized with total protein and tissue slice weight, respectively, and are presented as mean ± SEM. Number within bars represents the number of rat hearts in the group. * p < 0.01 vs. control and chymostatin groups.

Article Snippet: To determine the effects of the MC degranulation on SCF levels, LV slices from each heart were randomly separated into three groups with five or more slices per well: 1) culture media alone; 2) culture media with the MC secretagogue, compound 48/80 (20 μg/ml; Sigma, St Louis, MO); 3) culture media with compound 48/80 and anti-SCF; and 4) culture media with compound 48/80 and chymostatin (10 μM, MP Biomedicals, Solon, OH).

Techniques: Concentration Assay, Incubation

Journal: Frontiers in Immunology

Article Title: Annexin A1 Mimetic Peptide Ac 2-26 Modulates the Function of Murine Colonic and Human Mast Cells

doi: 10.3389/fimmu.2021.689484

Figure Lengend Snippet: Cytokine production by colon explants from C57BL/6 WT and C57BL/6 IL-4 KO mice treated in vivo with 1 mg/kg i.p. Ac 2-26 (+) or PBS (-). ELISA determined the concentration of IL-12 (A) , TNF-α (B) , IL-10 (C) and IL-4 (D) in explants stimulated for 24 h, with 20µg/mL compound 48/80 and with or without 3µM Ac2-26 treatment. Experiments were performed twice with n = 4 animals/group. Values of concentration in pg/mL are expressed as the mean ± S.D, n = 8. Data were analyzed using one-way ANOVA (post Tukey multiple-comparison test). **p <0.01 vs. respective WT group. ## p <0.01 vs. untreated in vivo and ex vivo (-/-) and && p <0.01 vs treated in vivo and not treated ex vivo (-/+).

Article Snippet: Remarkably, our study demonstrated that the AnxA1-derived peptide Ac 2-26 inhibited in vitro the degranulation of HMC-1 cells stimulated with compound 48/80 in FPR-dependent manner, as assessed by blocking the degranulation system using the antagonist BOC-2.

Techniques: In Vivo, Enzyme-linked Immunosorbent Assay, Concentration Assay, Ex Vivo

Journal: Frontiers in Immunology

Article Title: Annexin A1 Mimetic Peptide Ac 2-26 Modulates the Function of Murine Colonic and Human Mast Cells

doi: 10.3389/fimmu.2021.689484

Figure Lengend Snippet: Effect of peptide Ac2-26 on MC degranulation in colon explants. C57BL/6 WT and C57BL/6 interleukin-4- KO mice were pre-treated with PBS (-) or 3µM Ac2 -26 (+) and stimulated with 20µg/mL compound 48/80 ex vivo for 24 (h) (A) Representative colon sections with examples of intact (arrowheads) and degranulated (arrows) MCs. Staining: Toluidine Blue. Sections: 4μm. Scale bars: 20 μm. (B) Degranulation of MCs was assessed by an enzymatic assay. Experiments were performed twice with n = 4 animals/group. Values of the percentage of β hexosaminidase release are expressed as the mean ± S.D., n = 8. Data were analyzed using one-way ANOVA (post Tukey multiple-comparison test). **p <0.01 compared to respective WT group. ## p <0.01 compared to untreated in vivo and ex vivo (-/-) and && p <0.01 compared to treated and not treated ex vivo (-/+).

Article Snippet: Remarkably, our study demonstrated that the AnxA1-derived peptide Ac 2-26 inhibited in vitro the degranulation of HMC-1 cells stimulated with compound 48/80 in FPR-dependent manner, as assessed by blocking the degranulation system using the antagonist BOC-2.

Techniques: Ex Vivo, Staining, Enzymatic Assay, In Vivo

Journal: Frontiers in Immunology

Article Title: Annexin A1 Mimetic Peptide Ac 2-26 Modulates the Function of Murine Colonic and Human Mast Cells

doi: 10.3389/fimmu.2021.689484

Figure Lengend Snippet: Exogenous AnxA1 (peptide Ac2-26) inhibits HMC-1 cell line degranulation induced by compound 48/80. HMC-1 cells were pre-treated with 3, 5, or 10µM peptide Ac2-26 for 30 min in the absence or presence of FPRs antagonist BOC-2 (5µg/mL). Degranulation was assessed measuring the activity of β-hexosaminidase in culture supernatants. Results are expressed as percentage of enzyme release. All experiments were conducted in triplicate. Data are expressed as the mean ± S.D from four different experiments and analyzed using one-way ANOVA. ***p <0.001 compared to cells treated with 48/80 alone or 48/80 + Ac2-26 (3 or 5 µM); **p <0.01 compared to cells treated with 48/80 + Ac2-26 (10 µM) + BOC-2; ## p <0.01 compared to cells treated with 48/80 alone.

Article Snippet: Remarkably, our study demonstrated that the AnxA1-derived peptide Ac 2-26 inhibited in vitro the degranulation of HMC-1 cells stimulated with compound 48/80 in FPR-dependent manner, as assessed by blocking the degranulation system using the antagonist BOC-2.

Techniques: Activity Assay

Journal: Frontiers in Immunology

Article Title: Annexin A1 Mimetic Peptide Ac 2-26 Modulates the Function of Murine Colonic and Human Mast Cells

doi: 10.3389/fimmu.2021.689484

Figure Lengend Snippet: Exogenous AnxA1 (peptide Ac2-26) did not alter viability of HMC-1 cell line. Cells were pre-treated with 3, 5, or 10µM peptide Ac2-26 for 30 min in the absence or presence of FPRs antagonist BOC-2 (5µg/mL). The cells were then stimulated with 20 µg/mL compound 48/80 for 30 min. Cell viability was assessed by trypan blue exclusion assay and results are expressed as percentage. All experiments were performed in triplicate. Data are expressed as the mean ± S.D from four different experiments.

Article Snippet: Remarkably, our study demonstrated that the AnxA1-derived peptide Ac 2-26 inhibited in vitro the degranulation of HMC-1 cells stimulated with compound 48/80 in FPR-dependent manner, as assessed by blocking the degranulation system using the antagonist BOC-2.

Techniques: Trypan Blue Exclusion Assay

Journal: Cell Reports

Article Title: In Silico Modeling of Liver Metabolism in a Human Disease Reveals a Key Enzyme for Histidine and Histamine Homeostasis

doi: 10.1016/j.celrep.2016.05.014

Figure Lengend Snippet: Reduction of Histamine in PH1 Mice and Patients and in Cells Knocked Down for AGT (A and B) Histamine levels in livers (A) and sera (B) were reduced in Agxt −/− mice compared to wild-type (WT) mice (at least n = 3 per group). Agxt −/− mice with vector-mediated expression of AGT protein showed increased histamine in both livers (A) and serum (B). (C) Compared to WT mice, Agxt −/− showed reduced Evans blue extravasation in turned-over skin after intravenous injection of Evans blue followed by intradermal injections of either saline or compound 48/80. (D) The ratio of fluorescence units given by intradermal injection of 48/80 and normal saline was reduced in Agxt −/− mice (n = 8 per group). (E) AGT-knockdown Huh-7 cells (AGT KD, Figure S3 B) had reduced intracellular histamine compared to WT cells. (F) Plasma histamine levels were reduced in PH1 patients harboring AGXT mutations ( ; n = 3) compared to normal subjects (n = 5). (G and H) Histidine levels were reduced in livers (G) and sera (H) of Agxt −/− mice compared to WT. (I) Intraperitoneal injections of histidine resulted in increased histamine levels at 24 hr after the injections (T24) compared to baseline levels (T0) in both Agxt −/− and control WT mice (n = 6 per group; ∗ p < 0.05). See also , , and .

Article Snippet: Histamine-mediated skin permeability was determined through a gross staining using the compound 48/80 (Enzo Life Sciences) as described previously ( ).

Techniques: Plasmid Preparation, Expressing, Injection, Fluorescence