Journal: PLOS One

Article Title: In vivo mouse model of calcific myonecrosis induced by injury

doi: 10.1371/journal.pone.0346816

Figure Lengend Snippet: (a-c) Quantitative RT-PCR analysis of Nlrp3 (a) , Asc (b) , and Caspase-1 (c) mRNA expression in skeletal muscle from control and notexin-injected mice (n = 3). Data are presented as mean ± S.D. Statistical significance was assessed using unpaired t-test (** P < 0.01, *** P < 0.001). (d) Immunostaining of gastrocnemius muscle sections 3 days after notexin injection using anti-NLRP3, anti-ASC, and anti-Caspase-1 antibodies on serial sections. The upper panels show control muscles, and the lower panels show notexin-treated muscles. Scale bars, 20 μm. Asterisks indicate calcified muscle fibers. (e) Immunofluorescence staining showing the localization of NLRP3 (green), ASC or Caspase-1 (red) around calcified muscle fibers in notexin-injected skeletal muscle. Nuclei are counterstained with DAPI (blue). Scale bar, 20 μm. Asterisks indicate calcified muscle fibers. (f, g) Quantitative RT-PCR analysis of Il-1β (f) and Il-18 (g) mRNA expression in skeletal muscle from control and notexin-injected mice (n = 3). Data are presented as mean ± S.D. Statistical significance was assessed using unpaired t-test (*** P < 0.001). (h) Serum IL-18 levels measured by ELISA in control and notexin-injected mice (n = 6 (control), n = 9 (notexin)). Data are presented as mean ± S.D. Statistical significance was assessed using unpaired t-test (* P < 0.05). (i) Quantitative RT-PCR analysis of Runx2 mRNA expression in skeletal muscle from control and notexin-injected mice (n = 3). Data are presented as mean ± S.D. Statistical significance was assessed using unpaired t-test (* P < 0.05). (j) Western blot analysis of Runx2 and tubulin (loading control) protein expression in skeletal muscle from control and notexin-injected mice.

Article Snippet: Serum IL-18 concentrations were measured using the Mouse IL-18 DuoSet ELISA Kit (DY7625-05; R&D Systems) according to the manufacturer’s instructions.

Techniques: Quantitative RT-PCR, Expressing, Control, Injection, Immunostaining, Muscles, Immunofluorescence, Staining, Enzyme-linked Immunosorbent Assay, Western Blot

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