abx mixture  (MedChemExpress)

Journal: Life Metabolism

Article Title: Propionic acid secreted by Akkermansia muciniphila alleviates hepatic fibrosis by antioxidant regulation across the gut–liver axis

doi: 10.1093/lifemeta/loaf036

Figure Lengend Snippet: AKK alleviates liver fibrosis in mice by improving propionic acid metabolism. (a) Body weight of animals recorded weekly during the experimental period. (b) Liver index ( n = 6). (c–f) Serum content of ALT (c), AST (d), γ-GT (e), and ALP (f) ( n = 6). (g) Representative images of H&E, Masson, and Sirius Red staining (scale bar = 25 μm) in the liver ( n = 6). (h and i) Liver collagen deposition index ( n = 6). (j) Fibrosis-related markers such as TIMP-1, α-SMA, and Col1α1 and TGF-β1 protein expression assayed by western blotting. (k) Propionic acid levels in the livers, feces, and plasma of mice ( n = 6). (l) Gene expression of Slc52a1 and Slc52a2 in mice ( n = 6). Data are presented as the mean ± SEM. Statistical significance was determined using ANOVA for multiple-group comparisons. CCl 4 -induced hepatic fibrosis (CCl 4 ), ABX + CCl 4 , or a combination of ABX + CCl 4 plus AKK (ABX + CCl 4 + AKK ) were used to assess liver metabolomics and the indicated assays. ** P < 0.01, *** P < 0.05, compared with the CCl 4 group. # P < 0.05, ## P < 0.01, compared with the ABX + CCl 4 group. $ P < 0.05, compared with the CCl 4 + AKK group.

Article Snippet: To induce the ABX model, mice with hepatic fibrosis were orally administered an ABX mixture consisting of ampicillin (0.2 g/L, MedChemExpress, CAS# HY-B0522), vancomycin (0.2 g/L, MedChemExpress, CAS# HY-B0671), neomycin sulfate (0.2 g/L, MedChemExpress, CAS# HY-B0470), and metronidazole (0.2 g/L, MedChemExpress, CAS# HY-B0318) for 1 week before the 8th week.

Techniques: Staining, Expressing, Western Blot, Clinical Proteomics, Gene Expression

Journal: Pharmacological research

Article Title: Resveratrol-mediated attenuation of superantigen-driven acute respiratory distress syndrome is mediated by microbiota in the lungs and gut

doi: 10.1016/j.phrs.2021.105548

Figure Lengend Snippet: Effect of CMT from mice treated with RES or L. reuteri on SEB-mediated ARDS. Recipient mice were pre-treated with extensive regimen of 8 antibiotics (ABX) for four weeks followed by CMT from RES, VEH, SEB + RES, SEB + VEH donors or L. reuteri, and then exposed to SEB, as detailed in Methods. The various groups are represented as follows: CMT from Vehicle treated mice into ABX mice followed by SEB (VEH→SEB), CMT from RES treated mice into ABX mice followed by SEB (RES→SEB), CMT from SEB + VEH treated mice into ABX mice followed by SEB (SEB + VEH→SEB), L. reuteri transfer into ABX mice followed by SEB (L. Reuteri →SEB), and CMT from SEB + RES treated mice into ABX mice followed by SEB (SEB + RES→SEB). A) Cladogram shows the least significant discriminative changes in gut microbiota amongst various groups. B) Gut content assayed for SCFA including propionic and butyric acids. C) qPCR quantification of L. reuteri gene expression in bacterial DNA isolated from the gut. D) Infiltrating mononuclear cells (MNCs) counts in lung tissue. E-H) Flow cytometry results showing percentages of: E) Th1 in lungs. F) Th1 in the spleen G) NKT in the spleen and H) Th17 in the spleen. I) Survival rate observed for 30 days post-SEB exposure. One-way ANOVA test was used. Log-rank (Mantel-Cox) test was used to compare the survival curves. Bars denote Mean ± SEM and any significant differences (p < 0.05) are indicated by lowercase letters among groups.

Article Snippet: In order to deplete the endogenous microbiota, mice were treated for 4 weeks with the extensive antibiotic mixture (ABX) as described [ 34 ], containing 250 mg/ml bacitracin, 170 μg/ml gentamycin, 125 μg/ml ciprofloxacin, 100 μg/ml neomycin, 100 μg/ml metronidazole, 100 μg/ml ceftazidime, 100 U/ml penicillin, 50 μg/ml streptomycin and 50 μg/ml vancomycin (MilliporeSigma, St. Louis, MO, USA), supplemented with 1 g/L sweetener (Splenda, Carmel, IN, USA) to overcome the metallic taste of the mixture in drinking water.

Techniques: Gene Expression, Isolation, Flow Cytometry

Journal: Pharmacological research

Article Title: Resveratrol-mediated attenuation of superantigen-driven acute respiratory distress syndrome is mediated by microbiota in the lungs and gut

doi: 10.1016/j.phrs.2021.105548

Figure Lengend Snippet: Effect of CMT from mice treated with RES or L. reuteri on SEB-mediated ARDS. Recipient mice were pre-treated with extensive regimen of 8 antibiotics (ABX) for four weeks followed by CMT from RES, VEH, SEB + RES, SEB + VEH donors or L. reuteri, and then exposed to SEB, as detailed in Methods. The various groups are represented as follows: CMT from Vehicle treated mice into ABX mice followed by SEB (VEH→SEB), CMT from RES treated mice into ABX mice followed by SEB (RES→SEB), CMT from SEB + VEH treated mice into ABX mice followed by SEB (SEB + VEH→SEB), L. reuteri transfer into ABX mice followed by SEB (L. Reuteri →SEB), and CMT from SEB + RES treated mice into ABX mice followed by SEB (SEB + RES→SEB). A) Cladogram shows the least significant discriminative changes in gut microbiota amongst various groups. B) Gut content assayed for SCFA including propionic and butyric acids. C) qPCR quantification of L. reuteri gene expression in bacterial DNA isolated from the gut. D) Infiltrating mononuclear cells (MNCs) counts in lung tissue. E-H) Flow cytometry results showing percentages of: E) Th1 in lungs. F) Th1 in the spleen G) NKT in the spleen and H) Th17 in the spleen. I) Survival rate observed for 30 days post-SEB exposure. One-way ANOVA test was used. Log-rank (Mantel-Cox) test was used to compare the survival curves. Bars denote Mean ± SEM and any significant differences (p < 0.05) are indicated by lowercase letters among groups.

Article Snippet: Microbiota depletion and mouse treatment for CMT studies In order to deplete the endogenous microbiota, mice were treated for 4 weeks with the extensive antibiotic mixture (ABX) as described [ 34 ], containing 250 mg/ml bacitracin, 170 μg/ml gentamycin, 125 μg/ml ciprofloxacin, 100 μg/ml neomycin, 100 μg/ml metronidazole, 100 μg/ml ceftazidime, 100 U/ml penicillin, 50 μg/ml streptomycin and 50 μg/ml vancomycin (MilliporeSigma, St. Louis, MO, USA), supplemented with 1 g/L sweetener (Splenda, Carmel, IN, USA) to overcome the metallic taste of the mixture in drinking water.

Techniques: Gene Expression, Isolation, Flow Cytometry

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Modulating T follicular cells in vivo enhances antigen-specific humoral immunity

doi: 10.4049/jimmunol.2001434

Figure Lengend Snippet: Extended ABX treatment alters IgG subclass distribution during homeostasis. (A) CFUs grown on LB agar and BHI agar plates from fecal bacteria after antibiotic treatment of mice (n = 5 mice/group). (B) Stool samples sequenced using 16S data and analyzed to determine phylum level relative abundance. Serum Ig titers of WT (white filled) and ABX-treated mice (black filled) for (C) mIgG1 (n = 20–30), (D) mIgG2a/c (n = 20–30), (E) mIgG2b (n = 20–30), (F) mIgG3 (n = 20–30), (G) mIgE (n = 20–30), (H) mIgM (n = 10), (I) mIgA (n = 20), and (J) fecal mIgA (n = 10) determined by ELISA. Means and SEM (±SEM) are plotted. Results are representative of at least two independent repeats. The p values are for two-tailed unpaired Student t test were performed for statistical comparisons. *p < 0.05, **p < 0.01, ****p < 0.001. ns, not significant.

Article Snippet: The 5–6-wk-old C57BL/6 mice for antibiotic mixture (ABX) experiments and 7–8-wk-old C57BL/6 and NOD mice were purchased from The Jackson Laboratory and maintained in the animal facility at Massachusetts General Hospital under specific pathogen-free conditions according to the National Institutes of Health guidelines.

Techniques: Bacteria, Enzyme-linked Immunosorbent Assay, Two Tailed Test