Journal: bioRxiv

Article Title: Selective identification of the herb Picrorhiza kurroa and probiotic Lactobacillus fermentum as a synbiotic with fermentation-enhanced physicochemical, biological, and metabolomic properties

doi: 10.64898/2026.01.17.700043

Figure Lengend Snippet: Effect of different plant extracts on the proliferation of A-B Lactobacillus casei. C-D Lactobacillus fermentum. E-F Lactobacillus rhamnosus. *represents a significant difference at p<0.05 compared to the control.

Article Snippet: Three commercial LAB strains: Lactobacillus rhamnosus GG (LR) (ATCC 53103), Lactobacillus casei (LC) (ATCC 393), and Lactobacillus fermentum (LF) (ATCC 9338), and one Escherichia coli ( E. coli ) strain (ATCC 4157) were used in this study, which were procured from HiMedia Laboratory, Mumbai, India.

Techniques: Control

Journal: bioRxiv

Article Title: Selective identification of the herb Picrorhiza kurroa and probiotic Lactobacillus fermentum as a synbiotic with fermentation-enhanced physicochemical, biological, and metabolomic properties

doi: 10.64898/2026.01.17.700043

Figure Lengend Snippet: Effect of different plant extracts on the proliferation of A-B Escherichia coli. Prebiotic activity score (PAS) of plant extracts using C Lactobacillus casei. D Lactobacillus fermentum. E Lactobacillus rhamnosus. F Combined PAS using all LAB. G Effect of PK and AV on the proliferation of murine faecal LAB. *represents a significant difference compared to the control. *p<0.05, **p<0.01, ***p<0.001. Means that do not share a common letter indicate a significant difference at p<0.05.

Article Snippet: Three commercial LAB strains: Lactobacillus rhamnosus GG (LR) (ATCC 53103), Lactobacillus casei (LC) (ATCC 393), and Lactobacillus fermentum (LF) (ATCC 9338), and one Escherichia coli ( E. coli ) strain (ATCC 4157) were used in this study, which were procured from HiMedia Laboratory, Mumbai, India.

Techniques: Activity Assay, Control

Journal: Respiratory Research

Article Title: CSE triggers ferroptosis via SIRT4-mediated GNPAT deacetylation in the pathogenesis of COPD

doi: 10.1186/s12931-023-02613-0

Figure Lengend Snippet: Investigation of the association between the expression of plasmalogen biosynthesis and ferroptosis in COPD mice. ( A ) H&E staining of sections of mouse lung tissues from COPD model and normal group. ( B ) The concentration of IL-33 and IL-1α in the bronchoalveolar lavage fluid was determined by ELISA assay. ( C ) The neutrophils, macrophages, and lymphocytes in bronchoalveolar lavage fluid were quantified and compared between COPD and normal groups. ( D ) Tissue damage of lung were elevated by immunohistochemistry staingin of Bax. ( E ) GPX4 activity and the contents of GSH and MDA were analyzed between COPD and normal groups. ( F ) GPX4 expression in the lung tissues of mice was tested by immunohistochemistry (×100 and ×200). ( G ) The protein expression of GPX4, FAR-1, AGPS, and GNPAT was detected via western blot analysis. ( H ) The mRNA expression of FAR-1, AGPS, and GNPAT was measured via quantitative real-time PCR. ( I ) GNPAT expression in the lung tissues of mice was tested by immunohistochemistry (×100 and ×200). Data are presented as the mean ± SD of three replicates and analyzed using Student’s t-test. ns p > 0.05, ** p < 0.01, *** p < 0.001, compared with normal. The gels were cropped reasonably

Article Snippet: Subsequently, the sections were blocked with serum and subjected to incubation with primary antibodies targeting Bax (ab32503; Abcam), GPX4 (ab219592; Abcam) and GNPAT (14931-1-AP, Proteintech), which were diluted in PBS at 4 °C overnight.

Techniques: Expressing, Staining, Concentration Assay, Enzyme-linked Immunosorbent Assay, Immunohistochemistry, Activity Assay, Western Blot, Real-time Polymerase Chain Reaction

Journal: Respiratory Research

Article Title: CSE triggers ferroptosis via SIRT4-mediated GNPAT deacetylation in the pathogenesis of COPD

doi: 10.1186/s12931-023-02613-0

Figure Lengend Snippet: CSE triggered ferroptosis and upregulated the expression of plasmalogen biosynthesis in human alveolar epithelial cells. Human type II alveolar epithelial A549 cells were exposed to 0.1%, 0.5%, 2%, and 5% CSE for 24 h. ( A ) Cell viability was detected by CCK-8 assay. ( B ) LDH release was detected by LDH activity assays. ( C ) The concentration of IL-33 and IL-1α in the cellular supernatant was determined by ELISA assay. ( D ) The levels of total ROS were determined using a 2’, 7’-dichlorofluorescein diacetate (DCFH-DA) kit. ( E-G ) Quantification of the ferroptosis-related markers MDA, GSH, and GPX4. ( H ) The protein levels of GPX4, FAR-1, AGPS, and GNPAT were detected by western blot analysis. ( I ) The mRNA expression of FAR-1, AGPS, and GNPAT was measured via quantitative real-time PCR. Data are presented as the mean ± SD of three replicates and analyzed using one-way analysis of variance (ANOVA), followed by Dunnett’s test. ns p > 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001, compared with blank. The gels were cropped reasonably

Article Snippet: Subsequently, the sections were blocked with serum and subjected to incubation with primary antibodies targeting Bax (ab32503; Abcam), GPX4 (ab219592; Abcam) and GNPAT (14931-1-AP, Proteintech), which were diluted in PBS at 4 °C overnight.

Techniques: Expressing, CCK-8 Assay, Activity Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Real-time Polymerase Chain Reaction

Journal: Respiratory Research

Article Title: CSE triggers ferroptosis via SIRT4-mediated GNPAT deacetylation in the pathogenesis of COPD

doi: 10.1186/s12931-023-02613-0

Figure Lengend Snippet: Experimental validation of CSE-induced markers associated with ferroptosis and plasmalogen biosynthesis. ( A ) Mitochondrial morphology associated with ferroptosis was determined by transmission electron microscopy. ( B ) Content detection of the ferroptosis-related markers MDA, GSH, and GPX4 by ELISA. ( C ) The mRNA expression of GNPAT was measured via quantitative real-time PCR. ( D ) The protein levels of GPX4 and GNPAT were detected by western blot analysis. ( E ) Immunofluorescence staining of GNPAT was displayed in different groups. Data are presented as the mean ± SD of three replicates and analyzed using ANOVA, followed by Tukey’s post-hoc test. * p < 0.05, *** p < 0.001, compared with blank; $ p < 0.05, compared with 5% CSE; ## p < 0.01, ### p < 0.001, compared with 5% CSE + DMSO. The gels were cropped reasonably

Article Snippet: Subsequently, the sections were blocked with serum and subjected to incubation with primary antibodies targeting Bax (ab32503; Abcam), GPX4 (ab219592; Abcam) and GNPAT (14931-1-AP, Proteintech), which were diluted in PBS at 4 °C overnight.

Techniques: Biomarker Discovery, Transmission Assay, Electron Microscopy, Enzyme-linked Immunosorbent Assay, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Immunofluorescence, Staining

Journal: Respiratory Research

Article Title: CSE triggers ferroptosis via SIRT4-mediated GNPAT deacetylation in the pathogenesis of COPD

doi: 10.1186/s12931-023-02613-0

Figure Lengend Snippet: Interference with GNPAT can alleviate CSE-induced ferroptosis in human alveolar epithelial cells. A549 cells were transfected with sh-GNPAT or sh-NC for 48 h, followed by 5% CSE treatment. ( A-B ) The expression levels of GNPAT mRNA and protein were determined in the above A549 cells. ( C ) Immunofluorescence staining of GNPAT was shown. ( D ) Cell viability was detected by CCK-8 assay. ( E ) The LDH release was detected by LDH activity assays. ( F ) The levels of total ROS were determined using a 2’, 7’-dichlorofluorescein diacetate (DCFH-DA) kit. ( G ) Cell apoptosis was determined by flow cytometry assay. ( H ) Mitochondrial morphology associated with ferroptosis was determined by transmission electron microscopy. ( I ) Detection of lipid oxidation by the BODIPY 581/591 C11 probe method. ( J ) Content detection of the ferroptosis-related markers MDA, GSH, and GPX4 by ELISA. A549 cells were exposed to 0.1%, 0.5%, 2%, and 5% CSE for 24 h, followed by quantification of SIRT4 expression via western blot analysis ( K ) and GNPAT acetylation level by immunoprecipitation ( L ). Data are presented as the mean ± SD of three replicates and analyzed using Student’s t-test. *** p < 0.001, compared with sh-NC. The gels were cropped reasonably

Article Snippet: Subsequently, the sections were blocked with serum and subjected to incubation with primary antibodies targeting Bax (ab32503; Abcam), GPX4 (ab219592; Abcam) and GNPAT (14931-1-AP, Proteintech), which were diluted in PBS at 4 °C overnight.

Techniques: Transfection, Expressing, Immunofluorescence, Staining, CCK-8 Assay, Activity Assay, Flow Cytometry, Transmission Assay, Electron Microscopy, Enzyme-linked Immunosorbent Assay, Western Blot, Immunoprecipitation

Journal: Respiratory Research

Article Title: CSE triggers ferroptosis via SIRT4-mediated GNPAT deacetylation in the pathogenesis of COPD

doi: 10.1186/s12931-023-02613-0

Figure Lengend Snippet: Experimental validation of CSE-induced ferroptosis-related markers via SIRT4-mediated GNPAT acetylation. ( A ) Detection of lipid oxidation by the BODIPY 581/591 C11 probe method. ( B ) Mitochondrial morphology associated with ferroptosis was determined by transmission electron microscopy. ( C ) Content detection of the ferroptosis-related markers MDA, GSH, and GPX4 by ELISA. Data are presented as the mean ± SD of three replicates and analyzed using ANOVA, followed by Tukey’s post-hoc test. *** p < 0.001, compared with blank + vector; ### p < 0.001, compared with 5% CSE + vector; $$ p < 0.01, $$$ p < 0.001, compared with 5% CSE + SIRT4

Article Snippet: Subsequently, the sections were blocked with serum and subjected to incubation with primary antibodies targeting Bax (ab32503; Abcam), GPX4 (ab219592; Abcam) and GNPAT (14931-1-AP, Proteintech), which were diluted in PBS at 4 °C overnight.

Techniques: Biomarker Discovery, Transmission Assay, Electron Microscopy, Enzyme-linked Immunosorbent Assay, Plasmid Preparation