Journal: Frontiers in Microbiology

Article Title: Saccharomyces Boulardii Ameliorates Non-alcoholic Steatohepatitis in Mice Induced by a Methionine-Choline-Deficient Diet Through Gut-Liver Axis

doi: 10.3389/fmicb.2022.887728

Figure Lengend Snippet: SB reduced MCD diet-caused steatohepatitis in mice. C57BL/6 male mice were fed an oral control diet ( n = 10) or the MCD diet ( n = 18) and given vehicle or SB . (A) Representative liver sections after H & E staining, scale bars: 100μm. (B) Plasma levels of alanine aminotransferase (ALT). (C) Hepatic triglyceride content. (D) NAFLD activity score (NAS). (E) Parameters of NAS: Steatosis, inflammation and ballooning. (F) Representative sections of Trichrome Masson staining, scale bar: 50μm, and fibrosis score based on the NAS scoring system. * p < 0.05.

Article Snippet: The MCD diet (Mod TestDiet ® 5CC7 w/no added choline and methionine) was fed for eight weeks to induce liver injury ( n = 19).

Techniques: Control, Staining, Clinical Proteomics, Activity Assay

Journal: Frontiers in Microbiology

Article Title: Saccharomyces Boulardii Ameliorates Non-alcoholic Steatohepatitis in Mice Induced by a Methionine-Choline-Deficient Diet Through Gut-Liver Axis

doi: 10.3389/fmicb.2022.887728

Figure Lengend Snippet: Hepatic inflammatory responses caused by the MCD diet were suppressed by SB gavage. (A) Hepatic expressions of proinflammatory cytokines. (B) Representative liver sections of immunohistochemical staining of F4/80 and hepatic expression of F4/80. Scale bars: 50μm. * p < 0.05.

Article Snippet: The MCD diet (Mod TestDiet ® 5CC7 w/no added choline and methionine) was fed for eight weeks to induce liver injury ( n = 19).

Techniques: Immunohistochemical staining, Staining, Expressing

Journal: Frontiers in Microbiology

Article Title: Saccharomyces Boulardii Ameliorates Non-alcoholic Steatohepatitis in Mice Induced by a Methionine-Choline-Deficient Diet Through Gut-Liver Axis

doi: 10.3389/fmicb.2022.887728

Figure Lengend Snippet: SB ameliorated MCD diet-caused hepatic fibrosis in mice. (A) Representative sections of Sirus Red staining (Scale bar: 100μm) and hepatic c ollagen 1a1 expression. (B) Representative sections of immunohistochemical staining ofα-SMA (Scale bar: 50μm) and hepatic expression ofα- SMA. (C) Hepatic fibrotic gene expressions, Tissue Inhibitor of Metalloproteinase 1 ( Timp1) and Matrix Metallopeptidase 2 ( Mmp2) * p < 0.05.

Article Snippet: The MCD diet (Mod TestDiet ® 5CC7 w/no added choline and methionine) was fed for eight weeks to induce liver injury ( n = 19).

Techniques: Staining, Expressing, Immunohistochemical staining

Journal: Frontiers in Microbiology

Article Title: Saccharomyces Boulardii Ameliorates Non-alcoholic Steatohepatitis in Mice Induced by a Methionine-Choline-Deficient Diet Through Gut-Liver Axis

doi: 10.3389/fmicb.2022.887728

Figure Lengend Snippet: The MCD diet led to the shortening of the small intestine and intestinal villi, which were improved by the SB supplement. (A) Representative images of the small intestine of mice on the control diet ( n = 10) or the MCD diet ( n = 18), and also given vehicle or SB . The length of the small intestine improved after the SB supplement. (B) Representative sections of the distal small intestine after H & E staining. Both the intestine length of villi and the width of crypt improved after the SB supplement. Scale bars: 50μm. * p < 0.05.

Article Snippet: The MCD diet (Mod TestDiet ® 5CC7 w/no added choline and methionine) was fed for eight weeks to induce liver injury ( n = 19).

Techniques: Control, Staining

Journal: Frontiers in Microbiology

Article Title: Saccharomyces Boulardii Ameliorates Non-alcoholic Steatohepatitis in Mice Induced by a Methionine-Choline-Deficient Diet Through Gut-Liver Axis

doi: 10.3389/fmicb.2022.887728

Figure Lengend Snippet: SB ameliorated the MCD diet-caused intestinal inflammation. Intestinal expression of proinflammatory cytokines Il1b, Tnf -α, and CCL2 in the proximal, middle, distal small intestine (PSI, MSI, and DSI), and colon. * p < 0.05.

Article Snippet: The MCD diet (Mod TestDiet ® 5CC7 w/no added choline and methionine) was fed for eight weeks to induce liver injury ( n = 19).

Techniques: Expressing

Journal: Frontiers in Microbiology

Article Title: Saccharomyces Boulardii Ameliorates Non-alcoholic Steatohepatitis in Mice Induced by a Methionine-Choline-Deficient Diet Through Gut-Liver Axis

doi: 10.3389/fmicb.2022.887728

Figure Lengend Snippet: SB decreased the MCD diet-caused leaky gut with less Lipopolysaccharide translocation. (A) Fecal albumin measured by ELISA. (B) Plasma Lipopolysaccharide (LPS) level. (C) Intestinal expression of intestinal barrier gene, ZO-1 , in the proximal, middle and distal small intestine. (D) Representative liver sections of ZO-1 immunohistochemical staining of the distal small intestine. Scale bars: 50μm. * p < 0.05.

Article Snippet: The MCD diet (Mod TestDiet ® 5CC7 w/no added choline and methionine) was fed for eight weeks to induce liver injury ( n = 19).

Techniques: Translocation Assay, Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Expressing, Immunohistochemical staining, Staining

Journal: Frontiers in Microbiology

Article Title: Saccharomyces Boulardii Ameliorates Non-alcoholic Steatohepatitis in Mice Induced by a Methionine-Choline-Deficient Diet Through Gut-Liver Axis

doi: 10.3389/fmicb.2022.887728

Figure Lengend Snippet: SB affected the microbial configuration and restored intestinal microbiota diversity in the MCD diet-fed mice. (A) Topmost abundant taxa at the family level and (B) genus level. (C) The Shannon diversity index represents alpha diversity. (D) PCA represents beta diversity at the genus level. Each symbol represents one sample ( n = 5–10 per group). (E) Discriminative biomarkers with an LDA score > 4.0.

Article Snippet: The MCD diet (Mod TestDiet ® 5CC7 w/no added choline and methionine) was fed for eight weeks to induce liver injury ( n = 19).

Techniques:

Journal: Frontiers in Microbiology

Article Title: Saccharomyces Boulardii Ameliorates Non-alcoholic Steatohepatitis in Mice Induced by a Methionine-Choline-Deficient Diet Through Gut-Liver Axis

doi: 10.3389/fmicb.2022.887728

Figure Lengend Snippet: The MCD diet altered the fungal composition in the intestine of mice. (A) Topmost abundant taxa at the family and (B) genus level, MCD diet feeding showed increased Pichia and Trichosporon . Alpha diversity is represented by Shannon (C) and Simpson’s (D) diversity index. (E) Beta diversity presented by PCA. Each symbol represents one sample ( n = 4–6 per group).

Article Snippet: The MCD diet (Mod TestDiet ® 5CC7 w/no added choline and methionine) was fed for eight weeks to induce liver injury ( n = 19).

Techniques:

Journal: International Journal of Biological Sciences

Article Title: TM4SF5-mediated KEAP1 Regulation in Hepatocytes Irrelevant to NRF2 Expression and Activity Promotes Oxidative Stress and Inflammation to Develop Metabolic Dysfunction-Associated Steatotic Liver Disease

doi: 10.7150/ijbs.126251

Figure Lengend Snippet: Keap1 downregulation reverses TM4SF5-driven, MASH-associated fibrosis upon MCD diet. (A-D) WT, Alb -TG Tm4sf5-Flag , and Tm4sf5 -/- KO C57BL/6 male mice (n=4~5, 8-week-old) were maintained on an MCD diet for 3.5 weeks (A) and received intravenous injections of either PBS or siKeap1 (0.02 mg/kg, twice a week) prior to sacrifice for subsequent hepatic analyses, including body and liver weight measurements (B), and evaluation through H&E, Masson's trichrome (M-T), Oil Red O (ORO), or dihydroethidium (DHE) staining, immunohistochemistry (C), or immunoblotting (D) for specified proteins. Scale bar: 100 μm. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ***, P < 0.0001; ns, non-significant, one-way ANOVA. All data are expressed as mean ± SEM.

Article Snippet: For positive control groups in the MASH model, mice were fed an MCD diet (TD90262, Teklad) for 4 weeks or a MASH diet (A06071302, CDAHFD, Research Diet Inc., New Brunswick, NJ, USA) with L-amino acid rodent formulation containing 60 kcal% fat, 0.1% methionine, and without choline for 12 weeks.

Techniques: Staining, Immunohistochemistry, Western Blot

Journal: bioRxiv

Article Title: Flexibility of systemic one-carbon metabolism partially buffers dietary methyl donor deficiency

doi: 10.64898/2026.02.05.703880

Figure Lengend Snippet: (a) Schematic showing intravenous infusion of [trimethyl- H 9 ]choline and [methyl- C 1 ]methionine in mice fed a methionine and choline deficient diet or fed a paired control diet. (b) Rate of appearance of choline calculated from m+9 serum enrichments from 8 h intravenous infusion of [trimethyl- H 9 ]choline. (c) Rate of appearance of methionine calculated from m+1 serum enrichments from 8 h intravenous infusion of [methyl- C 1 ]methionine. (d) Normalized labeling of tissue SAM from 8 hour intravenous infusion of [methyl- C 1 ]methionine. (e) Normalized labeling of tissue SAM from 8 hour intravenous infusion of [trimethyl- H 9 ]choline. The red line represents the fraction of serum methionine that is labeled in control conditions and the green line represents the fraction of serum methionine that is labeled in MCD diet. Labeling below these lines indicates that labeling is potentially sourced from serum methionine pools rather than tissue autonomous production. (f) Normalized labeling of pooled PCs in the liver from 8 hour intravenous infusion of [trimethyl- H 9 ]choline. (g) Ratio of ion counts of pooled PC species to pooled PE species. Bars represent mean ± S.D. Each data point represents data from an independent mouse. P-values were calculated with two-way ANOVA with Šídák correction for multiple comparisons. All experiments were performed in male ad lib fed C57Bl/6N mice during the light cycle. PC, phosphatidylcholine; PE, phosphatidylethanoline; PEMT, phosphatidylethanolamine methyltransferase

Article Snippet: Methyl deficiency was induced by transitioning mice from standard chow to a methionine and choline-deficient (MCD) diet (MP Biomedicals, MP296043910), while age-matched controls received a matched methionine and choline replete control diet (MP Biomedicals, MP296044110).

Techniques: Control, Labeling

Journal: bioRxiv

Article Title: Flexibility of systemic one-carbon metabolism partially buffers dietary methyl donor deficiency

doi: 10.64898/2026.02.05.703880

Figure Lengend Snippet: (a) Normalized labeling of serum and tissue serine from 8 hour intravenous infusion of [trimethyl- H 9 ]choline.The red line represents the fraction of serum serine that is labeled in control conditions and the green line represents the fraction of serum serine that is labeled in MCD diet. Labeling below these lines indicates that labeling is potentially sourced from serum serine pools rather than tissue autonomous production. (b) Relative ion counts for glycine across tissues in MCD diet and control diet fed mice. Bars represent mean ± S.D. Each data point represents data from an independent mouse. P-values were calculated with two-way ANOVA with Šídák correction for multiple comparisons (a) or Mixed-effects analysis with Šídák correction for multiple comparisons (b). All experiments were performed in male ad lib fed C57Bl/6N mice during the light cycle. MCD, methionine and choline deficient

Article Snippet: Methyl deficiency was induced by transitioning mice from standard chow to a methionine and choline-deficient (MCD) diet (MP Biomedicals, MP296043910), while age-matched controls received a matched methionine and choline replete control diet (MP Biomedicals, MP296044110).

Techniques: Labeling, Control

Journal: bioRxiv

Article Title: Flexibility of systemic one-carbon metabolism partially buffers dietary methyl donor deficiency

doi: 10.64898/2026.02.05.703880

Figure Lengend Snippet: (a) Schematic of the folate cycle highlighting reactions that support production of 5,10-methylene-THF and SAM. (b) Schematic showing intravenous infusion of [2,3,3- H 3 ]serine and [U- C ]glycine in mice fed a methionine and choline deficient diet or fed a paired control diet. (c) Rate of appearance of serine calculated from m+3 serum enrichments from 8 h intravenous infusion of [2,3,3- H 3 ]serine. (d) Rate of appearance of glycine calculated from m+2 serum enrichments from 8 h intravenous infusion of [U- C 2 ]glycine. (e) Schematic of use of [2,3,3- H 3 ]serine tracing to measure the contribution of the folate cycle to SAM synthesis. (f) Normalized labeling of tissue SAM from 8 hour intravenous infusion of [2,3,3- H 3 ]serine. The red line represents the fraction of serum methionine that is labeled in control conditions and the green line represents the fraction of serum methionine that is labeled in MCD diet. Labeling below these lines indicates that labeling is potentially sourced from serum methionine pools rather than tissue autonomous production. Bars represent mean ± S.D. Each data point represents data from an independent mouse. P-values were calculated with two tailed T-test (c,d) or two-way ANOVA with Šídák correction for multiple comparisons (f). All experiments were performed in male ad lib fed C57Bl/6N mice during the light cycle. MCD, methionine and choline deficient; SHMT, serine hydroxymethyltransferase, MTR, 5-methyltetrahyrofolate homocysteine methyltransferase; SAM, s-adenosylmethionine

Article Snippet: Methyl deficiency was induced by transitioning mice from standard chow to a methionine and choline-deficient (MCD) diet (MP Biomedicals, MP296043910), while age-matched controls received a matched methionine and choline replete control diet (MP Biomedicals, MP296044110).

Techniques: Control, Labeling, Two Tailed Test

Journal: bioRxiv

Article Title: Flexibility of systemic one-carbon metabolism partially buffers dietary methyl donor deficiency

doi: 10.64898/2026.02.05.703880

Figure Lengend Snippet: (a) Schematic showing potential sources of circulating serine. (b) Normalized labeled fraction of serum serine from infusion of [U- C 2 ]glycine. (c) Normalized labeled fraction of serum serine from infusion of [U- C 6 ]glucose. (d) Rate of appearance of essential amino acid phenylalanine from intravenous infusion of [ N]phenylalanine. (e) Serine production fluxes from different sources measured using the serine rate of appearance and infusions of U- C 2 ]glycine, U- C 6 ]glucose, and [ N]phenylalanine. (f) Relative ion counts for serine across tissues in MCD diet and control diet fed mice. (g) Schematic of proposed mechanism. When methionine and choline are deficient in the diet, there is increased release of serine from the kidney that supports maintained methionine, PC, and choline synthesis in the liver. Thus, methionine and choline fluxes are maintained by one-carbon metabolism buffering. Bars represent mean ± S.D. Each data point represents data from an independent mouse. P-values were calculated with two tailed T-test (b), two-way ANOVA with Šídák correction for multiple comparisons (e), or mixed-effects analysis with Šídák correction for multiple comparisons (f). All experiments were performed in male ad lib fed C57Bl/6N mice during the light cycle. SAM, s-adenosylmethionine; MCD, methionine and choline deficient

Article Snippet: Methyl deficiency was induced by transitioning mice from standard chow to a methionine and choline-deficient (MCD) diet (MP Biomedicals, MP296043910), while age-matched controls received a matched methionine and choline replete control diet (MP Biomedicals, MP296044110).

Techniques: Labeling, Control, Two Tailed Test