factor κb ligand  (R&D Systems)

Journal: Synthetic and Systems Biotechnology

Article Title: Engineering Escherichia coli for robust Co-utilization of glucose and xylose enables high-titer succinate production from lignocellulosic hydrolysates

doi: 10.1016/j.synbio.2026.01.006

Figure Lengend Snippet: Investigation of xylose transport and PTS modification for succinate production. (A) Schematic representation of glucose and xylose transport routes in different E. coli strains, highlighting the key transporters and metabolic nodes influencing carbon flux; (B) Intracellular ATP levels in strains C600, MG1655, and BW25113 during aerobic growth on xylose; (C) Comparison of succinate and by-product accumulation between the parental strain C600 and engineered strain ESC2 under anaerobic conditions; (D) Fermentation performance of PTS-modified strain ESC3, showing sugar utilization, biomass generation, and succinate production; (E–F) Growth profiles of engineered ESC3 derivatives in defined medium with xylose (E) or glucose–xylose mixtures (F). All experimental data were performed in triplicate, and error bars represent the standard deviation. Statistical analysis was performed using a two-tailed Student's t -test (∗∗p < 0.01, ∗∗∗p < 0.001).

Article Snippet: In this study, we systematically engineered E. coli C600 (ATCC 23724) [ ], a strain with efficient and low-energy xylose transport, as the chassis for succinate production from lignocellulosic sugars.

Techniques: Modification, Comparison, Standard Deviation, Two Tailed Test

Journal: Synthetic and Systems Biotechnology

Article Title: Engineering Escherichia coli for robust Co-utilization of glucose and xylose enables high-titer succinate production from lignocellulosic hydrolysates

doi: 10.1016/j.synbio.2026.01.006

Figure Lengend Snippet: Construction of a succinate-producing strain from C600. (A) Metabolic map illustrating targeted knockouts ( ldhA , pflB , ptsG , adhE and pta-ackA ) and expression/integration of pck to redirect flux toward succinate; (B) Two-stage fermentation scheme comprising aerobic growth using shaking flasks and followed by anaerobic production in serum bottles; (C–D) Succinate fermentation of six engineered strains cultured on xylose (C) or glucose–xylose mixtures (D). All experimental data were performed in triplicate, and error bars represent the standard deviation.

Article Snippet: In this study, we systematically engineered E. coli C600 (ATCC 23724) [ ], a strain with efficient and low-energy xylose transport, as the chassis for succinate production from lignocellulosic sugars.

Techniques: Expressing, Cell Culture, Standard Deviation

Journal: Synthetic and Systems Biotechnology

Article Title: Engineering Escherichia coli for robust Co-utilization of glucose and xylose enables high-titer succinate production from lignocellulosic hydrolysates

doi: 10.1016/j.synbio.2026.01.006

Figure Lengend Snippet: Evaluation of exogenous xylose utilization pathways and library-based strain selection. (A) Schematic comparison of the endogenous XI pathway with the Dahms and Weimberg pathways; (B) Design of pathway plasmid libraries and RBS variants controlling expression of key genes for Dahms and Weimberg pathways. The Weimberg library plasmid carries XylA , XylX , and XylB from C. crescentus , while the Dahms library plasmid contains XylB from C. crescentus . The helper plasmid harbors xylC from C. crescentus and the endogenous yjhG from E. coli . RBS sequences were designed with 32 mutations, enabling gene expression levels ranging from 4 to 57,523 au; (C) Growth and succinate production of four representative ESC7 derivatives (ESC7-W1, ESC7-W2, ESC7-D1, ESC7-D2), which were randomly selected from the Weimberg (W1, W2) or Dahms (D1, D2) pathway libraries, compared with ESC6 (XI pathway); (D) Fermentation performance of the same four ESC7 clones carrying the helper plasmid (harboring XylC and yjhG ), compared with ESC6; (E) Validation of pathway combinations in the ESC6 background using the same four representative plasmids, integrating XI with Dahms/Weimberg routes and help plasmid; (F) Screening of library colonies identified six optimal variants, which were reconstructed in ESC6 and evaluated for succinate production from glucose–xylose mixtures. All experimental data were performed in triplicate, and error bars represent the standard deviation. Statistical analysis was performed using a two-tailed Student's t -test (∗∗∗ p < 0.001).

Article Snippet: In this study, we systematically engineered E. coli C600 (ATCC 23724) [ ], a strain with efficient and low-energy xylose transport, as the chassis for succinate production from lignocellulosic sugars.

Techniques: Selection, Comparison, Plasmid Preparation, Expressing, Gene Expression, Clone Assay, Biomarker Discovery, Standard Deviation, Two Tailed Test

Journal: Synthetic and Systems Biotechnology

Article Title: Engineering Escherichia coli for robust Co-utilization of glucose and xylose enables high-titer succinate production from lignocellulosic hydrolysates

doi: 10.1016/j.synbio.2026.01.006

Figure Lengend Snippet: Investigation of xylose transport and PTS modification for succinate production. (A) Schematic representation of glucose and xylose transport routes in different E. coli strains, highlighting the key transporters and metabolic nodes influencing carbon flux; (B) Intracellular ATP levels in strains C600, MG1655, and BW25113 during aerobic growth on xylose; (C) Comparison of succinate and by-product accumulation between the parental strain C600 and engineered strain ESC2 under anaerobic conditions; (D) Fermentation performance of PTS-modified strain ESC3, showing sugar utilization, biomass generation, and succinate production; (E–F) Growth profiles of engineered ESC3 derivatives in defined medium with xylose (E) or glucose–xylose mixtures (F). All experimental data were performed in triplicate, and error bars represent the standard deviation. Statistical analysis was performed using a two-tailed Student's t -test (∗∗p < 0.01, ∗∗∗p < 0.001).

Article Snippet: E. coli DH5α (Sangon Biotech) was used for gene cloning and plasmid construction.

Techniques: Modification, Comparison, Standard Deviation, Two Tailed Test

Journal: Synthetic and Systems Biotechnology

Article Title: Engineering Escherichia coli for robust Co-utilization of glucose and xylose enables high-titer succinate production from lignocellulosic hydrolysates

doi: 10.1016/j.synbio.2026.01.006

Figure Lengend Snippet: Evaluation of exogenous xylose utilization pathways and library-based strain selection. (A) Schematic comparison of the endogenous XI pathway with the Dahms and Weimberg pathways; (B) Design of pathway plasmid libraries and RBS variants controlling expression of key genes for Dahms and Weimberg pathways. The Weimberg library plasmid carries XylA , XylX , and XylB from C. crescentus , while the Dahms library plasmid contains XylB from C. crescentus . The helper plasmid harbors xylC from C. crescentus and the endogenous yjhG from E. coli . RBS sequences were designed with 32 mutations, enabling gene expression levels ranging from 4 to 57,523 au; (C) Growth and succinate production of four representative ESC7 derivatives (ESC7-W1, ESC7-W2, ESC7-D1, ESC7-D2), which were randomly selected from the Weimberg (W1, W2) or Dahms (D1, D2) pathway libraries, compared with ESC6 (XI pathway); (D) Fermentation performance of the same four ESC7 clones carrying the helper plasmid (harboring XylC and yjhG ), compared with ESC6; (E) Validation of pathway combinations in the ESC6 background using the same four representative plasmids, integrating XI with Dahms/Weimberg routes and help plasmid; (F) Screening of library colonies identified six optimal variants, which were reconstructed in ESC6 and evaluated for succinate production from glucose–xylose mixtures. All experimental data were performed in triplicate, and error bars represent the standard deviation. Statistical analysis was performed using a two-tailed Student's t -test (∗∗∗ p < 0.001).

Article Snippet: E. coli DH5α (Sangon Biotech) was used for gene cloning and plasmid construction.

Techniques: Selection, Comparison, Plasmid Preparation, Expressing, Gene Expression, Clone Assay, Biomarker Discovery, Standard Deviation, Two Tailed Test

Journal: eBioMedicine

Article Title: Autoantibodies against type I interferons in patients with zoonotic H7N9 influenza: an observational case–control study

doi: 10.1016/j.ebiom.2026.106387

Figure Lengend Snippet: Autoantibodies binding to IFNα 2 , IFNβ 1b and IFNω in patients with H7N9 infection and healthy controls. (A) Age and sex distribution of the three study groups. For each age group, the number of individuals positive for autoantibodies neutralising at least one tested IFN-I (IFNα 2 , IFNβ 1b , IFNω) at low concentrations is indicated in red. M, male; F, female; nAb+, positive for IFN-I-neutralising autoantibodies. (B) Detection of IgG autoantibodies binding to IFNα 2 , IFNβ 1b or IFNω in serum samples by multiplex bead-based assay. Samples with a Z-score >7 were considered positive for IFN-I-binding autoantibodies. Measurements were performed without technical replicates because of limited sample availability. (C) Prevalence of IFN-I-binding autoantibodies by IFN type and study group. IFNα 2 ± IFNβ 1b ± IFNω, positive for autoantibodies binding to at least one of the tested IFN-I; IFNα 2 + IFNω, positive for autoantibodies binding to both IFNα 2 and IFNω.

Article Snippet: Serum was diluted 1:50 in DMEM supplemented with 10% FCS and penicillin/streptomycin, and pre-incubated for 1 h at room temperature with one of the following IFN concentrations: (i) IFNα 2 (Novus Biologicals, NBP2-34971) at 10 ng/ml or 0.5 ng/ml; (ii) IFNβ 1b (PBL Assay Science, 11420-1) at 1 ng/ml or 0.25 ng/ml; (iii) IFN-ω (Novus Biologicals, NBP2-35893) at 10 ng/ml or 0.2 ng/ml.

Techniques: Binding Assay, Infection, Multiplex Assay, Bead-based Assay

Journal: eBioMedicine

Article Title: Autoantibodies against type I interferons in patients with zoonotic H7N9 influenza: an observational case–control study

doi: 10.1016/j.ebiom.2026.106387

Figure Lengend Snippet: Autoantibodies neutralising IFN-I in patients with H7N9 infection and healthy controls. (A) Luciferase-based reporter assay to assess the capacity of autoantibody positive sera to neutralise IFNα 2 (10 or 0.5 ng/ml), IFNβ 1b (1 or 0.25 ng/ml) or IFNω (10 or 0.2 ng/ml). Each sample was tested in biological duplicates and the mean values are shown. Samples were classified as neutralising if the mean of the relative luciferase activities was below 25% (dotted line) of the mean of the negative pool (four autoantibody-negative control sera). All sera positive for IFN-I-binding autoantibodies were tested; numbers are indicated above the graphs. Lines connect measurements of neutralising activity from the same serum sample at low and high IFN concentrations. (B) Prevalence of autoantibodies neutralising low IFN concentrations (IFNα 2 : 0.5 ng/ml, IFNβ 1b : 0.25 ng/ml; IFNω: 0.2 ng/ml) by IFN type and study group. IFNα 2 ± IFNβ 1b ± IFNω, positive for autoantibodies neutralising at least one tested IFN-I; IFNα 2 + IFNω, positive for autoantibodies neutralising both IFNα 2 and IFNω. (C) Area-proportional Venn diagrams illustrating the absolute numbers of samples with autoantibodies neutralising high and low concentrations of IFNα 2 (10 or 0.5 ng/ml), IFNβ 1b (1 or 0.25 ng/ml) or IFNω (10 or 0.2 ng/ml). Venn diagrams were created with BioVenn ( https://www.biovenn.nl/index.php ).

Article Snippet: Serum was diluted 1:50 in DMEM supplemented with 10% FCS and penicillin/streptomycin, and pre-incubated for 1 h at room temperature with one of the following IFN concentrations: (i) IFNα 2 (Novus Biologicals, NBP2-34971) at 10 ng/ml or 0.5 ng/ml; (ii) IFNβ 1b (PBL Assay Science, 11420-1) at 1 ng/ml or 0.25 ng/ml; (iii) IFN-ω (Novus Biologicals, NBP2-35893) at 10 ng/ml or 0.2 ng/ml.

Techniques: Infection, Luciferase, Reporter Assay, Negative Control, Binding Assay, Activity Assay

Journal: eBioMedicine

Article Title: Autoantibodies against type I interferons in patients with zoonotic H7N9 influenza: an observational case–control study

doi: 10.1016/j.ebiom.2026.106387

Figure Lengend Snippet: Association between the presence of IFN-I-neutralising autoantibodies and H7N9 infection. (A) The association between age, sex and IFN-I-neutralising autoantibodies in patients with H7N9 infection or in the two control groups combined (poultry workers + close contacts) was assessed using Firth's penalised logistic regression. Predicted probabilities for the presence of autoantibodies with 95% confidence intervals (CIs, shaded areas around the curve) are shown across participant age for men and women. To visualise the modelled probabilities in relation to the underlying data, we overlaid sex-specific age density distributions beneath the predicted probability curves. (B) Odds ratios (OR) with 95% CIs for the presence of autoantibodies neutralising low IFN concentrations in patients compared to healthy controls, adjusted for age and sex, determined by Firth’s penalised logistic regression models. See also for the results of the logistic regression analyses and for unadjusted estimates. IFNα 2 ± IFNω ± IFNβ 1b , positive for autoantibodies neutralising at least one of the tested IFN-I; IFNα 2 ± IFNω, positive for autoantibodies neutralising IFNα 2 and/or IFNω; ∗∗∗∗, p < 0.0001 (Firth’s penalised logistic regression).

Article Snippet: Serum was diluted 1:50 in DMEM supplemented with 10% FCS and penicillin/streptomycin, and pre-incubated for 1 h at room temperature with one of the following IFN concentrations: (i) IFNα 2 (Novus Biologicals, NBP2-34971) at 10 ng/ml or 0.5 ng/ml; (ii) IFNβ 1b (PBL Assay Science, 11420-1) at 1 ng/ml or 0.25 ng/ml; (iii) IFN-ω (Novus Biologicals, NBP2-35893) at 10 ng/ml or 0.2 ng/ml.

Techniques: Infection, Control

Journal: eBioMedicine

Article Title: Autoantibodies against type I interferons in patients with zoonotic H7N9 influenza: an observational case–control study

doi: 10.1016/j.ebiom.2026.106387

Figure Lengend Snippet: Neutralising sera block the antiviral effect of IFNα 2 in cell culture infected with IAV. Antiviral activity of IFNα 2 (5 ng/ml) against IAV (PR8-GFP, MOI 1) alone or in the presence of serially diluted IFN-I-neutralising sera (n = 19), autoantibody-negative sera (n = 4), or a monoclonal anti-IFNα 2 antibody in A549 cells. Infection rates (GFP + /DAPI + cells) at 7 h post-infection were normalised to untreated, infected cells. The dotted line indicates the reduction of infected cells after IFN treatment alone. If possible, the mean of two independent experiments is shown. Sufficient material was available for 12 out of 19 samples.

Article Snippet: Serum was diluted 1:50 in DMEM supplemented with 10% FCS and penicillin/streptomycin, and pre-incubated for 1 h at room temperature with one of the following IFN concentrations: (i) IFNα 2 (Novus Biologicals, NBP2-34971) at 10 ng/ml or 0.5 ng/ml; (ii) IFNβ 1b (PBL Assay Science, 11420-1) at 1 ng/ml or 0.25 ng/ml; (iii) IFN-ω (Novus Biologicals, NBP2-35893) at 10 ng/ml or 0.2 ng/ml.

Techniques: Blocking Assay, Cell Culture, Infection, Activity Assay