Journal: Frontiers in Immunology

Article Title: Removal of natural anti-αGal antibodies elicits protective immunity against Gram-negative bacterial infections

doi: 10.3389/fimmu.2023.1232924

Figure Lengend Snippet: Treatment with GAS914 impacts GalT-KO mice survival after CLP. (A) DNA stratification by pulsed-field gel electrophoresis (PFGE) of E coli isolated from mouse blood after 12 h – 24 h of CLP in GalT-KO mice treated with GAS914 (n = 8) or PBS (n = 8). (B) Influence of GAS914 or PBS treatment on GalT-KO mouse survival after CLP in animals treated before and after the procedure (left, n = 17) or beginning the treatment 12 h after CLP (right, n = 14). Data are represented in Kaplan-Maier curves and were compared using the long-rank (Mantel-Cox) test, **P < 0.01.

Article Snippet: We also measured in vitro GalT-KO mice serum bactericidal activity, and the antibody and complement deposition against the pathogenic mouse isolate E. coli A3 and E. coli O86:B7 (ATCC ® 12701 TM ), a bacterium with high α-galactosyl content ( , ).

Techniques: Pulsed-Field Gel, Electrophoresis, Isolation

Journal: Frontiers in Immunology

Article Title: Removal of natural anti-αGal antibodies elicits protective immunity against Gram-negative bacterial infections

doi: 10.3389/fimmu.2023.1232924

Figure Lengend Snippet: Treatment with GAS914 increases GalT-KO mice serum bactericidal capacity against E coli A3 isolated after CLP. (A) Effect of GAS914 and PBS in GalT-KO bactericidal activity against E coli A3. The bactericidal activity was calculated as the percentage of bacteria surviving in reaction mixtures containing the tested serum compared to the control (growth). Growth: control bacterial growth, ACP, alternative complement pathway; SMS, standard mouse serum. Individual data of PBS and GAS914 represent the mean of three experiments. (B) Median fluorescence intensity of IgG, IgM, C3, C4, and IgG subclasses on the surface of E coli A3 (n = 6). Individual data represents the mean of three experiments. Unpaired non-parametric Mann-Whitney test was used to compare PBS and GAS914 groups, ** P < 0.01.

Article Snippet: We also measured in vitro GalT-KO mice serum bactericidal activity, and the antibody and complement deposition against the pathogenic mouse isolate E. coli A3 and E. coli O86:B7 (ATCC ® 12701 TM ), a bacterium with high α-galactosyl content ( , ).

Techniques: Isolation, Activity Assay, Bacteria, Control, Fluorescence, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Removal of natural anti-αGal antibodies elicits protective immunity against Gram-negative bacterial infections

doi: 10.3389/fimmu.2023.1232924

Figure Lengend Snippet: Neutralizing anti-αGal antibodies from human sera with GAS914 increases serum bactericidal killing and decreases IgG2 binding to Gram-negative bacteria. (A) Bactericidal killing after exposing the human sera to PBS or GAS914 against E coli O86:B7, P. aeruginosa 21565, and K pneumonia 35204 (n = 8). The bactericidal killing was calculated as the percentage of the number of bacteria surviving in reaction mixtures containing the tested serum compared to the control (growth). Growth: control bacterial growth, ACP, alternative complement pathway. Individual data of PBS and GAS914 represent the mean of three experiments. (B) Median fluorescence intensity of IgG, IgM, IgA, IgG subclasses, and complement C3 and C4 deposition on the surface of E coli O86:B7 (n = 8). Individual data represents the mean of three experiments. Comparisons were analyzed by paired t -tests, * P < 0.05, ** P < 0.01.

Article Snippet: We also measured in vitro GalT-KO mice serum bactericidal activity, and the antibody and complement deposition against the pathogenic mouse isolate E. coli A3 and E. coli O86:B7 (ATCC ® 12701 TM ), a bacterium with high α-galactosyl content ( , ).

Techniques: Binding Assay, Bacteria, Control, Fluorescence

Journal: Frontiers in Immunology

Article Title: Removal of natural anti-αGal antibodies elicits protective immunity against Gram-negative bacterial infections

doi: 10.3389/fimmu.2023.1232924

Figure Lengend Snippet: Length and expression level of lipopolysaccharide (LPS) O-antigen chains and αGal in Gram-negative bacteria. (A) Separation by SDS-PAGE (13%) of repeating oligosaccharide units of LPS extracted from Escherichia coli O86:B7 (lane 1), Pseudomonas aeruginosa 21565 (lane 2), Klebsiella pneumoniae 35204 (lane 3) and Escherichia coli A3 (lane 4). Standard LPS from E coli O111:B4 (lane 5, Sigma-Aldrich, St. Louis, MO, USA) was used as a reference. (B) Median fluorescence intensity of α-galactosyl expression with isolectin IB4 and αGal expression with anti-αGal IgM and IgG monoclonal antibodies in E coli A3, K pneumoniae 35204, P. aeruginosa 21565, and E coli O86:B7. Data are representative of three independent experiments.

Article Snippet: We also measured in vitro GalT-KO mice serum bactericidal activity, and the antibody and complement deposition against the pathogenic mouse isolate E. coli A3 and E. coli O86:B7 (ATCC ® 12701 TM ), a bacterium with high α-galactosyl content ( , ).

Techniques: Expressing, Bacteria, SDS Page, Fluorescence, Bioprocessing

Journal: Pharmaceutics

Article Title: Mucosal Adhesive Chitosan Nanogel Formulations of Antibiotics and Adjuvants (Terpenoids, Flavonoids, etc.) and Their Potential for the Treatment of Infectious Diseases of the Gastrointestinal Tract.

doi: 10.3390/pharmaceutics15092353

Figure Lengend Snippet: Figure 3. The dependences of E. coli and Lactobacillus colony-forming units on the incubation time of cells with antibacterial drugs. For E. coli: C(LF) = 1 µg/mL, C(MN) = 0.1 mg/mL, C(zephirol) = 0.1 mg/mL, C(EG) = 0.1 mg/mL. For Lactobacillus: C(LF) = 10 µg/mL, C(MN) = 1 mg/mL, C(zephirol) = 0.1 mg/mL, C(EG) = 1 mg/mL. LB medium (pH 7.2). 37 ◦C.

Article Snippet: The experimental steps necessary for the development of such a composition are given below: (1) obtaining inclusion complexes of antibiotics and adjuvants with MCD, as well as the inclusion of these complexes in nanogels of chitosan, cross-linked or not with genipin; (2) primary screening of antibacterial activity of formulations against E. coli ATCC 25922 (as a screening model of pathogenic bacteria) and Lactobacilli (as a model of normal microbiome) to optimize the composition of the drug; (3) studying the effect of the most promising preparations in a long-term experiment; (4) the study of the mechanisms of antibacterial action of nanogels using FTIR spectroscopy, which provides information about the structural components of cells, as well as the study of the physicochemical basis of the nanogels formation; (5) study of antioxidant and mucoadhesive properties of drugcontaining chitosan nanogels, which are important for preventing bacterial infection and stimulating early healing; (6) studies of the activity of the effect of drugs on sturgeon fish; (7) summarizing the activity of drug formulations in vitro and in vivo and discussing the potential applicability for the treatment of infectious diseases of the gastrointestinal tract.

Techniques: Incubation

Journal: Pharmaceutics

Article Title: Mucosal Adhesive Chitosan Nanogel Formulations of Antibiotics and Adjuvants (Terpenoids, Flavonoids, etc.) and Their Potential for the Treatment of Infectious Diseases of the Gastrointestinal Tract.

doi: 10.3390/pharmaceutics15092353

Figure Lengend Snippet: Figure 4. (a) FTIR spectra of suspension of E. coli cells (109 CFU) after a day of incubation with drug formulations in Chit5 particles. C(LF) = 1 µg/mL, C(MN, MM, SHA) = 0.1 mg/mL, C (other substances) = 1 mg/mL. LB medium (pH 7.2). (b) FTIR spectra of suspension of E. coli cells (109 CFU) during incubation (online) with linalool–MCD in Chit5 particles. 37 ◦C. (c,d) Flow cytometry dia- grams of E. coli cells incubated with FITC-labeled Chit5–genipin nanogel 15 min (CFITC = 1 µg/mL). (e,f) Flow cytometry diagrams of E. coli cells (control). Green indicates a population with a high inten- sity of FITC fluorescence. SSC—side scattering, FSC—front scattering, FITC—fluorescence channel.

Article Snippet: The experimental steps necessary for the development of such a composition are given below: (1) obtaining inclusion complexes of antibiotics and adjuvants with MCD, as well as the inclusion of these complexes in nanogels of chitosan, cross-linked or not with genipin; (2) primary screening of antibacterial activity of formulations against E. coli ATCC 25922 (as a screening model of pathogenic bacteria) and Lactobacilli (as a model of normal microbiome) to optimize the composition of the drug; (3) studying the effect of the most promising preparations in a long-term experiment; (4) the study of the mechanisms of antibacterial action of nanogels using FTIR spectroscopy, which provides information about the structural components of cells, as well as the study of the physicochemical basis of the nanogels formation; (5) study of antioxidant and mucoadhesive properties of drugcontaining chitosan nanogels, which are important for preventing bacterial infection and stimulating early healing; (6) studies of the activity of the effect of drugs on sturgeon fish; (7) summarizing the activity of drug formulations in vitro and in vivo and discussing the potential applicability for the treatment of infectious diseases of the gastrointestinal tract.

Techniques: Suspension, Incubation, Flow Cytometry, Labeling, Control

Journal: Frontiers in Pharmacology

Article Title: A Comprehensive Account on Recent Progress in Pharmacological Activities of Benzimidazole Derivatives

doi: 10.3389/fphar.2021.762807

Figure Lengend Snippet: Recently published patents of benzimidazole derivatives.

Article Snippet: A number of substituted fluorobenzimidazole derivatives ( 329–330 ) were synthesized and evaluated for in vitro antimycobacterial property against pathogenic M. tuberculosis H37Rv strain (ATCC 27294) using MABA method.

Techniques: Inhibition, Synthesized, Virus

Journal: Nature Communications

Article Title: Living plastics from plasticizer-assisted thermal molding of silk protein

doi: 10.1038/s41467-024-55097-x

Figure Lengend Snippet: a Schematic of oral delivery of REcN encapsulated in silk plastics for enhanced probiotic viability through the digestive system. b Photograph of WS/REcN and schematic illustrating REcN embedded within a protective, high-crystallinity silk shell (Scale bar: 5 mm). c SEM images of silk/REcN powders (Scale bar: 500 nm). d SEM images of WS/REcN (left) and individual REcN embedded in silk (right) (Scale bars: 5 μm left, 500 nm right). e Remaining viability of REcN from silk/REcN, TS/REcN, and WS/REcN ( n = 3 independent replicates, P = 0.3421 (silk/REcN vs. TS/REcN), P = 0.3261 (silk/REcN vs. WS/REcN), P = 0.9991 (TS/REcN vs. WS/REcN)). f Remaining viability of S-REcN (SGF + SIF-treated REcN), ST-REcN (REcN released from SGF + SIF-treated TS/REcN), and SW-REcN (REcN released from SGF + SIF-treated WS/REcN) ( n = 5 independent replicates, P ≤ 0.0001 (SW-REcN vs. ST-REcN), P ≤ 0.0001 (SW-REcN vs. S-REcN), P = 0.6245 (S-REcN vs. ST-REcN)). g Growth curves of fresh REcN, S-REcN, SW-REcN, and W-REcN (REcN released from untreated WS/REcN), measured by OD600 over 12 h ( n = 3 independent replicates, P ≤ 0.0001 (S-REcN vs. REcN or SW-REcN or W-REcN)). h Fluorescence microscopy images of SW-REcN and S-REcN (Green: SYTO-9, Red: RFP, Scale bar: 100 μm). i Schematic of in vitro antimicrobial activity of REcN and SW-REcN against Shigella flexneri ( S. flexneri ), with pathogen reduction quantified by plating on MacConkey agar ( n = 5 independent replicates, P = 0.364 (SW-REcN vs. REcN)). j In vivo probiotic resistance studies: orally administered REcN or WS/REcN with equivalent bacterial loads, with gastrointestinal tract imaging at 4 and 24 h post-administration ( n = 5 biological independent replicates). All data are mean values ± standard deviations. Statistical analysis: One-way ANOVA with Tukey’s multiple comparisons in ( e – g ) and Two-tailed unpaired t -test in ( i ) (**** P ≤ 0.0001, NS: not significant). i , j Created with BioRender.com released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license.

Article Snippet: Fresh REcN and SW-REcN (REcN released from SGF + SIF-treated WS/REcN) were collected to assess their antimicrobial activity against the gut pathogen Shigella flexneri ( S. flexneri , 12022 GFP, ATCC, USA).

Techniques: Fluorescence, Microscopy, In Vitro, Activity Assay, In Vivo, Imaging, Two Tailed Test