Journal: Microorganisms

Article Title: Saccharomyces cerevisiae TAD1 Mutant Strain As Potential New Antimicrobial Agent: Studies on Its Antibacterial Activity and Mechanism of Action

doi: 10.3390/microorganisms13122848

Figure Lengend Snippet: Growth curve measurement. ( A ) Loss of TAD1 does not impair S. cerevisiae growth. Growth curves of parental strain BY4743, S. boulardii , and S. cerevisiae TAD1-KO cultivated in YPD at 30 °C, 200 rpm for 24 h. The superimposable curves indicate that TAD1 deletion is growth-neutral under standard laboratory conditions. ( B – D ) S. cerevisiae TAD1-KO elicits potent metabolite-mediated growth suppression. Growth curves of E. coli , S. aureus , and S. typhi exposed to 90% ( v / v ) S. cerevisiae TAD1-KO , BY4743, or nutrient controls at 37 °C, 200 rpm. S. cerevisiae TAD1-KO CFS significantly delayed lag-to-log transition, reduced µmax, and lowered stationary-phase density. Equivalent growth in sterile water and YPD controls excludes nutrient limitation, confirming inhibition by secreted bioactive metabolites. Data are means ± SD (n = 3); ** p < 0.01; *** p < 0.001; ns denotes no statistically significant difference.

Article Snippet: Even at 25% ( v / v ), CFS reduced the CV-stainable biomass below the untreated control, while 90% CFS suppressed biofilm formation to ≤16% for all tested strains: E. coli ATCC 25922 (10.5%), E. coli mcr-1 (11.1%), S. aureus ATCC 29213 (9.0%), MRSA (10.8%), and S. typhi SL1344 (15.9%).

Techniques: Sterility, Inhibition

Journal: Microorganisms

Article Title: Saccharomyces cerevisiae TAD1 Mutant Strain As Potential New Antimicrobial Agent: Studies on Its Antibacterial Activity and Mechanism of Action

doi: 10.3390/microorganisms13122848

Figure Lengend Snippet: S. cerevisiae TAD1-KO CFS inhibits biofilm formation and downregulates biofilm-associated genes: ( A ) Quantitative CV assay showing dose-dependent suppression of biofilms in E. coli ATCC25922, E. coli ( mcr-1 ), S. aureus ATCC29213, MRSA, and S. typhi SL1344 after 24 h exposure to 25–90% ( v / v ) S. cerevisiae TAD1-KO CFS. ( B ) 96-well plate images confirming progressive biofilm disruption with increasing CFS concentration. ( C ) Light-microscopy images confirming progressive biofilm disruption with increasing CFS concentration. ( D ) qRT-PCR demonstrating significant downregulation of key biofilm genes ( fimH , fliC , csgA , csgD ) in E. coli treated with 50% S. cerevisiae TAD1-KO CFS versus untreated control. Data are means ± SD (n = 3); * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Even at 25% ( v / v ), CFS reduced the CV-stainable biomass below the untreated control, while 90% CFS suppressed biofilm formation to ≤16% for all tested strains: E. coli ATCC 25922 (10.5%), E. coli mcr-1 (11.1%), S. aureus ATCC 29213 (9.0%), MRSA (10.8%), and S. typhi SL1344 (15.9%).

Techniques: Disruption, Concentration Assay, Light Microscopy, Quantitative RT-PCR, Control

Journal: Microorganisms

Article Title: Saccharomyces cerevisiae TAD1 Mutant Strain As Potential New Antimicrobial Agent: Studies on Its Antibacterial Activity and Mechanism of Action

doi: 10.3390/microorganisms13122848

Figure Lengend Snippet: S. cerevisiae TAD1-KO CFS impairs adhesion, hydrophobicity, and EPS production: ( A ) Adhesion to glass after 24 h, ( B ) cell-surface hydrophobicity, and ( C ) EPS yield of E. coli , S. aureus , and S. typhi following 24 h exposure to 50% ( v / v ) S. cerevisiae TAD1-KO CFS. All parameters were significantly reduced versus untreated controls, confirming that CFS targets the early physical determinants of biofilm formation. Data are means ± SD (n = 3); * p < 0.05; ** p < 0.01; *** p < 0.001.

Article Snippet: Even at 25% ( v / v ), CFS reduced the CV-stainable biomass below the untreated control, while 90% CFS suppressed biofilm formation to ≤16% for all tested strains: E. coli ATCC 25922 (10.5%), E. coli mcr-1 (11.1%), S. aureus ATCC 29213 (9.0%), MRSA (10.8%), and S. typhi SL1344 (15.9%).

Techniques: Cell Surface Hydrophobicity

Journal: Microorganisms

Article Title: Saccharomyces cerevisiae TAD1 Mutant Strain As Potential New Antimicrobial Agent: Studies on Its Antibacterial Activity and Mechanism of Action

doi: 10.3390/microorganisms13122848

Figure Lengend Snippet: Time–kill kinetics and ultrastructural damage induced by S. cerevisiae TAD1-KO CFS. ( A ) Viable counts of E. coli exposed to 90% S. cerevisiae TAD1-KO CFS, 90% BY4743 CFS, or LB at 37 °C; bactericidal effect could be achieved within 4 h, with maximum bactericidal activity at 8 h. ( B ) Representative TEM micrographs after 6 h treatment with 50% S. cerevisiae TAD1-KO CFS: E. coli and S. aureus exhibit swelling, membrane rupture, cytoplasmic leakage, and plasmolysis compared with intact untreated cells, confirming metabolite-mediated envelope disruption. The black scale in the image represents 2 µm and 500 nm. Data are means ± SD (n = 3); *** p < 0.001.

Article Snippet: Even at 25% ( v / v ), CFS reduced the CV-stainable biomass below the untreated control, while 90% CFS suppressed biofilm formation to ≤16% for all tested strains: E. coli ATCC 25922 (10.5%), E. coli mcr-1 (11.1%), S. aureus ATCC 29213 (9.0%), MRSA (10.8%), and S. typhi SL1344 (15.9%).

Techniques: Activity Assay, Membrane, Disruption

Journal: Microorganisms

Article Title: Saccharomyces cerevisiae TAD1 Mutant Strain As Potential New Antimicrobial Agent: Studies on Its Antibacterial Activity and Mechanism of Action

doi: 10.3390/microorganisms13122848

Figure Lengend Snippet: S. cerevisiae TAD1-KO CFS sequentially disrupts envelope integrity and elicits intracellular ROS. ( A ) Time-dependent leakage of periplasmic AKP (OD 520 ) after 50% S. cerevisiae TAD1-KO CFS exposure; significant increase from 4 h onward. ( B ) NPN fluorescence assay: 50% S. cerevisiae TAD1-KO CFS raises outer-membrane permeability to the level of 1 mg mL −1 EDTA-2Na (positive control). ( C ) ONPG hydrolysis (OD 415 ), indicating inner-membrane permeabilization; slightly higher but statistically different from control (* p < 0.05). ( D ) DCFH-DA fluorescence: 90% S. cerevisiae TAD1-KO CFS triggers robust intracellular ROS accumulation (*** p < 0.001), contributing to rapid bacterial killing. ( E , F ) ROS scavengers do not impair S. cerevisiae TAD1-KO CFS bactericidal efficacy. E. coli (1 × 10 7 CFU mL −1 ) were cultured with 90% S. cerevisiae TAD1-KO CFS alone or supplemented with 1 mmol L −1 or 2 mmol L −1 VC, thiourea, or both. Viable counts (CFU mL −1 ) were determined at 0, 4 and 8 h (37 °C, 200 rpm). No statistically significant difference between groups ( p > 0.05) indicates that ROS contributes minimally to CFS-mediated killing; the lethal effect is primarily attributable to direct envelope disruption. Data are means ± SD (n = 3); * p < 0.05; ** p < 0.01; *** p < 0.001; ns denotes no statistically significant difference.

Article Snippet: Even at 25% ( v / v ), CFS reduced the CV-stainable biomass below the untreated control, while 90% CFS suppressed biofilm formation to ≤16% for all tested strains: E. coli ATCC 25922 (10.5%), E. coli mcr-1 (11.1%), S. aureus ATCC 29213 (9.0%), MRSA (10.8%), and S. typhi SL1344 (15.9%).

Techniques: Fluorescence, Membrane, Permeability, Positive Control, Control, Cell Culture, Disruption

Journal: Microorganisms

Article Title: Saccharomyces cerevisiae TAD1 Mutant Strain As Potential New Antimicrobial Agent: Studies on Its Antibacterial Activity and Mechanism of Action

doi: 10.3390/microorganisms13122848

Figure Lengend Snippet: S. cerevisiae TAD1-KO tolerates simulated gastrointestinal stress and displays superior aggregation traits. ( A ) Survival after 3 h in simulated gastric fluid (pH 3.0, pepsin 3 mg mL −1 ); S. cerevisiae TAD1-KO retained 86.0% viability. ( B ) Survival after 4 h in intestinal fluid (pH 8.0, trypsin 1 mg mL −1 , 0.3% bile); 80.7% cells remained viable. ( C ) Auto-aggregation at 5 h; S. cerevisiae TAD1-KO exceeded BY4743 by 23%. ( D ) Co-aggregation increments with E. coli , S. aureus , and S. typhi (28%, 17%, and 30% higher than BY4743, respectively). Data are means ± SD (n = 3); ** p < 0.01; *** p < 0.001; ns denotes no statistically significant difference.

Article Snippet: Even at 25% ( v / v ), CFS reduced the CV-stainable biomass below the untreated control, while 90% CFS suppressed biofilm formation to ≤16% for all tested strains: E. coli ATCC 25922 (10.5%), E. coli mcr-1 (11.1%), S. aureus ATCC 29213 (9.0%), MRSA (10.8%), and S. typhi SL1344 (15.9%).

Techniques:

Journal: Microorganisms

Article Title: Saccharomyces cerevisiae TAD1 Mutant Strain As Potential New Antimicrobial Agent: Studies on Its Antibacterial Activity and Mechanism of Action

doi: 10.3390/microorganisms13122848

Figure Lengend Snippet: G. mellonella toxicity and therapeutic efficacy of S. cerevisiae TAD1-KO . ( A ) Survival curves after a single injection of live yeast, heat-killed yeast, or CFS (n = 5 per group); no larval mortality within 5 days confirms low toxicity. ( B , C ) Models of infection with E. coli and S. aureus : live S. cerevisiae TAD1-KO and its CFS significantly extended survival (*** p < 0.001 vs. PBS), whereas heat-inactivated yeast had no effect, demonstrating that protection requires metabolically active cells. ( D ) Survival rates of G. mellonella in each group on the first day of the E. coli infection model and the S. aureus infection model. ( E , F ) On the fifth day post-infection, homogenize G. mellonella and calculate the bacterial load within each group. ( G ) After homogenization, each group was spread onto LB agar plates for counting. ** p < 0.01; *** p < 0.001; ns denotes no statistically significant difference.

Article Snippet: Even at 25% ( v / v ), CFS reduced the CV-stainable biomass below the untreated control, while 90% CFS suppressed biofilm formation to ≤16% for all tested strains: E. coli ATCC 25922 (10.5%), E. coli mcr-1 (11.1%), S. aureus ATCC 29213 (9.0%), MRSA (10.8%), and S. typhi SL1344 (15.9%).

Techniques: Drug discovery, Injection, Infection, Metabolic Labelling, Homogenization

Journal: Microorganisms

Article Title: Saccharomyces cerevisiae TAD1 Mutant Strain As Potential New Antimicrobial Agent: Studies on Its Antibacterial Activity and Mechanism of Action

doi: 10.3390/microorganisms13122848

Figure Lengend Snippet: ( A – C ) Direct co-culture reveals enhanced pathogen suppression by live S. cerevisiae TAD1-KO . Viable counts (CFU mL −1 ) of E. coli , S. aureus , and S. typhi after 16 h contact with live S. cerevisiae TAD1-KO or BY4743 at 37 °C, 200 rpm. Inhibition rates: 89.5%, 55.1%, and 52.2% for S. cerevisiae TAD1-KO versus ≤ 20% for BY4743, confirming the contribution of TAD1 deletion to antimicrobial competence. Data are means ± SD; * p < 0.05; ** p < 0.01; *** p < 0.001; ns denotes no statistically significant difference.

Article Snippet: Even at 25% ( v / v ), CFS reduced the CV-stainable biomass below the untreated control, while 90% CFS suppressed biofilm formation to ≤16% for all tested strains: E. coli ATCC 25922 (10.5%), E. coli mcr-1 (11.1%), S. aureus ATCC 29213 (9.0%), MRSA (10.8%), and S. typhi SL1344 (15.9%).

Techniques: Co-Culture Assay, Inhibition

Journal: Microorganisms

Article Title: Saccharomyces cerevisiae TAD1 Mutant Strain As Potential New Antimicrobial Agent: Studies on Its Antibacterial Activity and Mechanism of Action

doi: 10.3390/microorganisms13122848

Figure Lengend Snippet: Transwell assay confirms metabolite-mediated antibacterial activity of S. cerevisiae TAD1-KO . ( A , B ) Viable counts of E. coli and S. aureus after 16 h at 37 °C. Inhibition rates: mixed culture 81.7%, separated culture 83.7%; no significant difference between regimes ( p > 0.05, n = 3), demonstrating that diffusible metabolites, not direct contact, drive antibacterial activity. Data are means ± SD; ** p < 0.01; *** p < 0.001; ns denotes no statistically significant difference.

Article Snippet: Even at 25% ( v / v ), CFS reduced the CV-stainable biomass below the untreated control, while 90% CFS suppressed biofilm formation to ≤16% for all tested strains: E. coli ATCC 25922 (10.5%), E. coli mcr-1 (11.1%), S. aureus ATCC 29213 (9.0%), MRSA (10.8%), and S. typhi SL1344 (15.9%).

Techniques: Transwell Assay, Activity Assay, Inhibition

Journal: Microorganisms

Article Title: Saccharomyces cerevisiae TAD1 Mutant Strain As Potential New Antimicrobial Agent: Studies on Its Antibacterial Activity and Mechanism of Action

doi: 10.3390/microorganisms13122848

Figure Lengend Snippet: Antimicrobial activity of S. cerevisiae TAD1-KO is pH-dependent. Inhibitory efficacy against E. coli and S. aureus (OD 600 , 24 h) declined sharply as the native CFS (pH 4.0 ± 0.37) was stepwise neutralized to pH 7.0, demonstrating that the bactericidal effect is mediated predominantly by acidic metabolites identified in the metabolomic analysis. Data are means ± SD; *** p < 0.001; ns denotes no statistically significant difference.

Article Snippet: Even at 25% ( v / v ), CFS reduced the CV-stainable biomass below the untreated control, while 90% CFS suppressed biofilm formation to ≤16% for all tested strains: E. coli ATCC 25922 (10.5%), E. coli mcr-1 (11.1%), S. aureus ATCC 29213 (9.0%), MRSA (10.8%), and S. typhi SL1344 (15.9%).

Techniques: Activity Assay

Journal: Microorganisms

Article Title: Saccharomyces cerevisiae TAD1 Mutant Strain As Potential New Antimicrobial Agent: Studies on Its Antibacterial Activity and Mechanism of Action

doi: 10.3390/microorganisms13122848

Figure Lengend Snippet: Mechanism diagram of S. cerevisiae TAD1-KO against E. coli . S. cerevisiae TAD1-KO CFS secretes large quantities of organic acids, causing a reduction in the expression of biofilm-associated genes in E. coli (blue arrow); CFS disrupts the normal cellular structure of E. coli , leading to increased leakage of AKP (blue arrow) and NPN (pink arrow) into the extracellular space, alongside elevated intracellular ONP levels (yellow arrow). Concurrently, organic acids promote intracellular ROS accumulation (red arrow), inducing oxidative stress damage within the cell.

Article Snippet: Even at 25% ( v / v ), CFS reduced the CV-stainable biomass below the untreated control, while 90% CFS suppressed biofilm formation to ≤16% for all tested strains: E. coli ATCC 25922 (10.5%), E. coli mcr-1 (11.1%), S. aureus ATCC 29213 (9.0%), MRSA (10.8%), and S. typhi SL1344 (15.9%).

Techniques: Expressing

Journal: Nanomaterials

Article Title: Microbial Nanotechnology: Challenges and Prospects for Green Biocatalytic Synthesis of Nanoscale Materials for Sensoristic and Biomedical Applications

doi: 10.3390/nano10010011

Figure Lengend Snippet: Nanomaterials synthesized by bacteria.

Article Snippet: Acetobacter xylinus GIM1.327 , Static culture in polysaccharides enriched medium, 30 °C (120 h) , Bacterial nanocellulose nanofibrils impregnated with Ag-NPs , Nanoporous three-dimensional network structure with a random arrangement of ribbon-shaped microfibrils without any preferential orientation; 2 to 100 nm (Ag NPs) , Intracellular-extracellular synthesis via enzymes glucokinase, phosphoglucomutase, UDPG, pyro-phospho-rylase and cellulose synthase , In vitro pH-responsive antimicrobial activity against Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 6538, Bacillus subtilis ATCC 9372 and Candida albicans CMCC(F) 98001 , [ , ] .

Techniques: Synthesized, Bacteria, Cell Culture, Antioxidant Activity Assay, Activity Assay, Virus, In Vitro, Sterility, Incubation, Sonication, Suspension, Fluorescence, Imaging, Dispersion, Inhibition, Membrane, Genetically Modified, Modification, Expressing, In Vivo, Luciferase

Journal: Nanomaterials

Article Title: Microbial Nanotechnology: Challenges and Prospects for Green Biocatalytic Synthesis of Nanoscale Materials for Sensoristic and Biomedical Applications

doi: 10.3390/nano10010011

Figure Lengend Snippet: Nanomaterials synthesized by microalgae.

Article Snippet: Acetobacter xylinus GIM1.327 , Static culture in polysaccharides enriched medium, 30 °C (120 h) , Bacterial nanocellulose nanofibrils impregnated with Ag-NPs , Nanoporous three-dimensional network structure with a random arrangement of ribbon-shaped microfibrils without any preferential orientation; 2 to 100 nm (Ag NPs) , Intracellular-extracellular synthesis via enzymes glucokinase, phosphoglucomutase, UDPG, pyro-phospho-rylase and cellulose synthase , In vitro pH-responsive antimicrobial activity against Escherichia coli ATCC 25922, Staphylococcus aureus ATCC 6538, Bacillus subtilis ATCC 9372 and Candida albicans CMCC(F) 98001 , [ , ] .

Techniques: Synthesized, Modification, Membrane, Control, Activity Assay, Suspension, Purification, Clinical Proteomics, Derivative Assay, Functional Assay, Raman Spectroscopy, Concentration Assay, Chromatography, Thin Layer Chromatography, Sterility, Biomarker Discovery

Journal: Pharmaceuticals

Article Title: Antimicrobial Effect of Honey Phenolic Compounds against E. coli —An In Vitro Study

doi: 10.3390/ph17050560

Figure Lengend Snippet: Effect of honey phenolic compounds on the growth of E. coli (ATCC 25922). Results are expressed as the mean SEM of log 10 cfu/mL. Experimental data for each concentration of phenolic compounds, methanol/sterile water solvent (20%/80%), and penicillin–streptomycin (2000U penicillin; 2 mg/mL streptomycin) solvent were compared to the TSB growth control. * Results are significantly different from TSB control; ** sesult for PLA at 7 mg/mL concentration; *** phloretin at 2 mg/mL concentration was associated with the exceed growth of E. coli in all rounds of the experiment. TSB—tryptone soy broth; PCA—3-phenyllactic acid; PCA— p -coumaric acid; Phl—phloretin.

Article Snippet: Lu et al. (2013) investigated the antibacterial characteristics of Manuka and Kanuka honeys, Manuka–Kanuka blends, and clover honey against E. coli O157:H7, B. subtilis 168, S. aureus ATCC 25923, and P. aeruginosa PAO1 (ATCC 15692) using a broth microdilution assay.

Techniques: Concentration Assay, Sterility, Solvent, Control

Journal: Pharmaceuticals

Article Title: Antimicrobial Effect of Honey Phenolic Compounds against E. coli —An In Vitro Study

doi: 10.3390/ph17050560

Figure Lengend Snippet: Effect of penicillin–streptomycin solvent on the growth of E. coli (ATCC 25922) in combination with p -coumaric acid. Results are expressed as the mean with the SEM of zone of inhibition after incubation at 37 °C for 24 h. Experimental data for p -coumaric acid at 2 mg/mL concentration and methanol/sterile water solvent (20%/80%) were compared to the TSB growth control. * Results are significantly different from TSB control.

Article Snippet: Lu et al. (2013) investigated the antibacterial characteristics of Manuka and Kanuka honeys, Manuka–Kanuka blends, and clover honey against E. coli O157:H7, B. subtilis 168, S. aureus ATCC 25923, and P. aeruginosa PAO1 (ATCC 15692) using a broth microdilution assay.

Techniques: Solvent, Inhibition, Incubation, Concentration Assay, Sterility, Control

Journal: Pharmaceuticals

Article Title: Antimicrobial Effect of Honey Phenolic Compounds against E. coli —An In Vitro Study

doi: 10.3390/ph17050560

Figure Lengend Snippet: Key studies regarding the antimicrobial effects of some phenolic compounds against E. coli .

Article Snippet: Lu et al. (2013) investigated the antibacterial characteristics of Manuka and Kanuka honeys, Manuka–Kanuka blends, and clover honey against E. coli O157:H7, B. subtilis 168, S. aureus ATCC 25923, and P. aeruginosa PAO1 (ATCC 15692) using a broth microdilution assay.

Techniques: Produced, Bacteria, Inhibition, Activity Assay, Diffusion-based Assay, Binding Assay, Flow Cytometry, Membrane, Fluorescence

Journal: Journal of Insect Science

Article Title: Isolation, Characterization, Kinetics, and Enzymatic and Nonenzymatic Microbicidal Activities of a Novel c-Type Lysozyme from Plasma of Schistocerca gregaria (Orthoptera: Acrididae)

doi: 10.1093/jisesa/iev038

Figure Lengend Snippet: Purification of lysozyme from plasma of fifth instar Schistocerca gregaria with cation-exchange chromatography on CM-Sepharose Fast Flow ion-exchange column. Proteins obtained with 60% (NH 4 ) 2 SO 4 saturation of hemolymph fifth instars S. gregaria immunized with pricking by a thin needle previously dipped into a concentrated pellet (2 × 10 8 cells/ml) of Escherichia coli were applied to column (20 × 2 cm) equilibrated with 0.5 M Na-acetate (pH 6.5) buffer. Elution was carried out with a gradient of 0.5–3.0 M of the same buffer at a flow rate of 1 ml/min (——, A 280 ;…….., Na-acetate concentration; ○- - - -○, anti- Micrococcus lysodeikticus activity). Five microliter of the fraction samples were used for the inhibition zone assay. Active fractions (anti- M. lysodeikticus fractions), tubes 31–39, were pooled, desalted, lyophilized, and stored at −20°C for further use.

Article Snippet: Escherichia coli (ATCC 11775) , 24 , – , 12 , 5.

Techniques: Purification, Clinical Proteomics, Chromatography, Concentration Assay, Activity Assay, Inhibition

Journal: Journal of Insect Science

Article Title: Isolation, Characterization, Kinetics, and Enzymatic and Nonenzymatic Microbicidal Activities of a Novel c-Type Lysozyme from Plasma of Schistocerca gregaria (Orthoptera: Acrididae)

doi: 10.1093/jisesa/iev038

Figure Lengend Snippet: Antimicrobial activities of purified Sg l against different bacteria, yeasts, and filamentous fungi

Article Snippet: Escherichia coli (ATCC 11775) , 24 , – , 12 , 5.

Techniques: Purification, Bacteria, Inhibition

Journal: Journal of Insect Science

Article Title: Isolation, Characterization, Kinetics, and Enzymatic and Nonenzymatic Microbicidal Activities of a Novel c-Type Lysozyme from Plasma of Schistocerca gregaria (Orthoptera: Acrididae)

doi: 10.1093/jisesa/iev038

Figure Lengend Snippet: Kinetics of permeabilization of the inner membrane (in parallel to antibacterial activity) by the Sg l against E. coli ML-35 p. The release of β-galactosidase from the lactose permease-deficient E. coli ML-35 p under the effect of the Sg l in an incubation medium and monitored by tracing the production of o -nitrophenol over time (1, 2, 3, and 4 h post incubation) from the specific substrate o -nitrophenyl-β-D-galactopyranoside. The absorbance (which is proportional to the β-galactosidase activity) was plotted against the incubation time. Values shown represent the mean of three experiments. In the control, an equivalent volume of sterile ddw replaced the Sg l solution. HEWL was used as a standard reference.

Article Snippet: Escherichia coli (ATCC 11775) , 24 , – , 12 , 5.

Techniques: Membrane, Activity Assay, Incubation, Control, Sterility

Journal: Frontiers in Microbiology

Article Title: Boosting Synergistic Effects of Short Antimicrobial Peptides With Conventional Antibiotics Against Resistant Bacteria

doi: 10.3389/fmicb.2021.747760

Figure Lengend Snippet: The minimal inhibitory concentrations (MICs) of S1, S1-Nal, S1-Nal-Nal, vancomycin, ciprofloxacin, and tetracycline.

Article Snippet: Antimicrobial peptide CLP-19 displayed synergistic effects with antibiotics such as ampicillin, ceftazidime, and levofloxacin against E. coli ATCC 25922 and S. aureus ATCC 29213 ( ).

Techniques:

Journal: Frontiers in Microbiology

Article Title: Boosting Synergistic Effects of Short Antimicrobial Peptides With Conventional Antibiotics Against Resistant Bacteria

doi: 10.3389/fmicb.2021.747760

Figure Lengend Snippet: The membrane permeabilization of Enterococcus faecium BCRC 15B0132, Acinetobacter baumannii BCRC 14B0091, A. baumannii BCRC 14B0097, A. baumannii BCRC 14B0100, Escherichia coli BCRC 13B0198, and E. coli BCRC 13B0207, by S1, S1-Nal, and S1-Nal-Nal for 60 min exposure. Calcein-AM loaded cells (10 7 CFU/ml) were resuspended by PBS, and the aliquots of 100 μl were added into a sterile black-wall 96-well plate, then treated with 0.5 × MIC of peptides in each well and measured the calcein leakage immediately. Each time point for the peptides was repeated three times independently.

Article Snippet: Antimicrobial peptide CLP-19 displayed synergistic effects with antibiotics such as ampicillin, ceftazidime, and levofloxacin against E. coli ATCC 25922 and S. aureus ATCC 29213 ( ).

Techniques: Membrane, Sterility

Journal: Frontiers in Microbiology

Article Title: Boosting Synergistic Effects of Short Antimicrobial Peptides With Conventional Antibiotics Against Resistant Bacteria

doi: 10.3389/fmicb.2021.747760

Figure Lengend Snippet: Synergistic effects of S1, S1-Nal, and S1-Nal-Nal in combination with antibiotics against bacterial strains studied.

Article Snippet: Antimicrobial peptide CLP-19 displayed synergistic effects with antibiotics such as ampicillin, ceftazidime, and levofloxacin against E. coli ATCC 25922 and S. aureus ATCC 29213 ( ).

Techniques:

Journal: Frontiers in Microbiology

Article Title: Boosting Synergistic Effects of Short Antimicrobial Peptides With Conventional Antibiotics Against Resistant Bacteria

doi: 10.3389/fmicb.2021.747760

Figure Lengend Snippet: S1-Nal and S1-Nal-Nal increase the uptake of BODIPY-labeled vancomycin in E. coli . (A) Fluorescence images of 10 7 CFU/ml E. coli BCRC 13B0207 treated with 0.5 × MIC of S1, S1-Nal, and S1-Nal-Nal at 37°C for 30 min, then treated BODIPY-labeled vancomycin for 30 mins (scale bar represents 5 μm). (B) Mean fluorescence intensity of S1, S1-Nal, and S1-Nal-Nal treated in E. coli BCRC 13B0207. Samples treated with BODIPY-labeled vancomycin only served as a control. Results are presented as means ± SEM, *** P < 0.001 compared with control.

Article Snippet: Antimicrobial peptide CLP-19 displayed synergistic effects with antibiotics such as ampicillin, ceftazidime, and levofloxacin against E. coli ATCC 25922 and S. aureus ATCC 29213 ( ).

Techniques: Labeling, Fluorescence, Control

Journal: Frontiers in Microbiology

Article Title: Boosting Synergistic Effects of Short Antimicrobial Peptides With Conventional Antibiotics Against Resistant Bacteria

doi: 10.3389/fmicb.2021.747760

Figure Lengend Snippet: Attenuation of LPS release from E. coli . Escherichia coli BCRC 13B0198 were incubated at the mid-log phase (10 4 CFU/ml) and treated with peptide alone (at 1 × MIC) or combination with vancomycin (both at 0.5 × MIC) at 37°C for 6 h. The samples were filtered through 0.2 μm pore filter and the endotoxin level was detected by LAL assay. *** P < 0.001 compared with vancomycin only. ns, no significant differences compared with vancomycin only.

Article Snippet: Antimicrobial peptide CLP-19 displayed synergistic effects with antibiotics such as ampicillin, ceftazidime, and levofloxacin against E. coli ATCC 25922 and S. aureus ATCC 29213 ( ).

Techniques: Incubation, LAL Assay

Journal: Biochemical Pharmacology

Article Title: A novel real time imaging platform to quantify macrophage phagocytosis

doi: 10.1016/j.bcp.2016.07.011

Figure Lengend Snippet: Fluorescent object segmentation analysis by the IncuCyte Basic Software. BMDMs (10 5 ) from male C57BL/6J mice were incubated with 1 mg/ml (A) or 10 μg/ml (B) green E. coli bioparticles and fluorescence emission was measured in the IncuCyte imaging platform for one hour at 37 °C. Representative images showing the green channel signal (left), the green object mask which was applied for quantification (middle) and the overlay of the two (right). (C) The edge split setting in the IncuCyte Basic Software allows for quantification of closely-spaced objects in both fluorescence channels. Images from 100,000 BMDMs incubated with 1 mg/ml green E. coli bioparticles for one hour at 37 °C without (left) or with (right) edge split. The magenta lines delineate separate objects. Scale bar 100 μm. BMDMs (2.5 × 10 4 per well) were incubated with an equal mix of 200 μg/ml red pHrodo S. aureus bioparticles and 200 μg/ml FITC-conjugated Zymosan A (D) or FITC-conjugated 2 μm latex beads (E) and phagocytosis was studied at one hour using confocal microscopy. White arrows indicate FITC-conjugated Zymosan A or latex beads on the macrophage membrane, orange arrows indicate unphagocytosed FITC-conjugated Zymosan A or latex beads and yellow arrows indicate unphagocytosed S. aureus bioparticles. NucBlue Live ReadyProbes reagent was used for nuclei staining in (D). Images are from one representative of three independent experiments. Scale bar 20 μm.

Article Snippet: Sterile IncuCyte™ green pHrodo® E. coli and Zymosan were purchased from ESSEN Bioscience (Welwyn Garden City, UK).

Techniques: Software, Incubation, Fluorescence, Imaging, Confocal Microscopy, Membrane, Staining

Journal: Biochemical Pharmacology

Article Title: A novel real time imaging platform to quantify macrophage phagocytosis

doi: 10.1016/j.bcp.2016.07.011

Figure Lengend Snippet: BMDM phagocytosis is pathogen particle number and cell density-dependent. (A) BMDMs (10 5 ) from male C57BL/6J mice were incubated with green E. coli bioparticles (1 mg/ml to 1 μg/ml) and fluorescence emission was measured in the IncuCyte imaging platform every 10 min for one hour at 37 °C. (B)–(D) Representative images from one experiment at one hour. (E) BMDMs (100,000–1562 cells per well) were incubated with 200 μg/ml green E. coli bioparticles for one hour at 37 °C and fluorescence emission was measured in the IncuCyte imaging platform with 10 min intervals. (F) RAW264.7, BV-2, THP-1 and hiPS cell-derived macrophages (5 × 10 4 ) were incubated with 200 μg/ml green E. coli bioparticles and fluorescence emission was measured in the IncuCyte imaging platform every 10 min for one hour at 37 °C. (G) Side-by-side comparison of green E. coli bioparticle phagocytosis by 100,000 BMDMs measured with a spectrofluorometer and the IncuCyte system. Data are presented as mean ± SEM from 3 to 4 biological replicates in technical duplicates, apart from F and G where data are from one experiment with two technical replicates and are presented as mean ± SD. Scale bar 100 μm.

Article Snippet: Sterile IncuCyte™ green pHrodo® E. coli and Zymosan were purchased from ESSEN Bioscience (Welwyn Garden City, UK).

Techniques: Incubation, Fluorescence, Imaging, Derivative Assay, Comparison

Journal: Biochemical Pharmacology

Article Title: A novel real time imaging platform to quantify macrophage phagocytosis

doi: 10.1016/j.bcp.2016.07.011

Figure Lengend Snippet: Macrophage phagocytosis is inhibited by actin and microtubule inhibitors and enhanced by opsonization. (A)–(D) BMDMs (5 × 10 4 ) from C57BL/6J male mice were pre-treated with the actin and microtubule inhibitors Cytochalasin D and Nocodazole at the indicated concentration for one hour and were then incubated with 200 μg/ml green E. coli bioparticles at 37 °C for another hour. (E) and (F) BMDMs (5 × 10 4 ) from C57BL/6J male mice were pre-treated with the vacuolar H + -ATPase inhibitor Bafilomycin A1 at the indicated concentration for one hour and were incubated with 200 μg/ml green E. coli bioparticles at 37 °C for another hour. (G) and (H) 200 μg/ml green E. coli bioparticles were opsonized with murine IgG or diluted human serum for 30 min at 37 °C and were then administered to 50,000 BMDMs from C57BL/6J male mice for one hour. (A), (C), (E) and (G) demonstrate the kinetics of phagocytosis at 10 min intervals. Fluorescence emission from all experiments was measured in the IncuCyte imaging platform and statistical analysis was carried out with one-way ANOVA with Dunnett’s multiple comparisons post hoc test. Data are presented as mean ± SEM from 3 to 5 biological replicates in technical duplicates.

Article Snippet: Sterile IncuCyte™ green pHrodo® E. coli and Zymosan were purchased from ESSEN Bioscience (Welwyn Garden City, UK).

Techniques: Concentration Assay, Incubation, Fluorescence, Imaging

Journal: Biochemical Pharmacology

Article Title: A novel real time imaging platform to quantify macrophage phagocytosis

doi: 10.1016/j.bcp.2016.07.011

Figure Lengend Snippet: Primary murine macrophage populations exhibit differential efficiency of phagocytosis of different pathogens. C57BL/6J murine lungs and peritoneal cavities were lavaged and cells were lysed to remove red blood cells. (A) Alveolar macrophages were defined as Siglec F + and CD11c + cells within the CD45 + cell population of total lung cells. (B) Peritoneal resident macrophages were defined as F4/80 + and CD11b + cells within the CD45 + cell population of total peritoneal cavity cells. (C) and (D) Macrophage numbers were inferred from total numbers and the Flow Cytometry gating strategy. (E) Alveolar, peritoneal and BMDMs (12,500 cells) were treated with 200 μg/ml green sterile Zymosan, E. coli and red S. aureus bioparticles for four hours and fluorescence emission was measured in the IncuCyte imaging platform every 15 min. (F)–(H) Kinetics of pathogen phagocytosis; 12,500 alveolar (F), peritoneal (G) and murine BMDMs (H) over four hours measured in the IncuCyte platform every 15 min. Data at four hours are presented as mean ± SEM from 4 biological replicates in technical duplicates. Statistical analysis was carried out with a two-way ANOVA and Tukey’s post hoc multiple comparisons test.

Article Snippet: Sterile IncuCyte™ green pHrodo® E. coli and Zymosan were purchased from ESSEN Bioscience (Welwyn Garden City, UK).

Techniques: Flow Cytometry, Sterility, Fluorescence, Imaging

Journal: Biochemical Pharmacology

Article Title: A novel real time imaging platform to quantify macrophage phagocytosis

doi: 10.1016/j.bcp.2016.07.011

Figure Lengend Snippet: Long term E. coli phagocytosis is biphasic, whereas S. aureus uptake reaches a plateau at 4 h. (A) and (B) BMDMs from male C57BL/6J mice were seeded at 50,000, 12,500 and 3125 cells per well and incubated with 200 μg/ml green E. coli (A) or red S. aureus (B) bioparticles. Fluorescence signal was measured in the IncuCyte imaging platform every 20 min for 24 h at 37 °C. Data are presented as mean ± SEM from 3 biological replicates in technical duplicates.

Article Snippet: Sterile IncuCyte™ green pHrodo® E. coli and Zymosan were purchased from ESSEN Bioscience (Welwyn Garden City, UK).

Techniques: Incubation, Fluorescence, Imaging

Journal: Biochemical Pharmacology

Article Title: A novel real time imaging platform to quantify macrophage phagocytosis

doi: 10.1016/j.bcp.2016.07.011

Figure Lengend Snippet: Macrophage polarization status affects phagocytic capacity. (A) BMDMs (2 × 10 5 ) from C57BL/6J male mice were polarized by incubation with vehicle, 100 ng/ml LPS, 20 ng/ml IFN-γ, 20 ng/ml IL-4 or 10 ng/ml IL-10 at 37 °C for 16 h and (A) Nos2 , (B) Arg1 and (C) Mrc1 mRNA expression was measured. (D) Polarized BMDMs (5 × 10 4 ) from C57BL/6J male mice were subsequently incubated with 200 μg/ml green E. coli bioparticles for one hour. Fluorescence emission was measured in the IncuCyte imaging platform for one hour every 10 min. (E) Time kinetics of the macrophage polarization experiments. Data are presented as mean ± SEM from 4 biological replicates in technical duplicates. Statistical analysis was carried out with one-way ANOVA with Dunnett’s multiple comparisons post hoc test.

Article Snippet: Sterile IncuCyte™ green pHrodo® E. coli and Zymosan were purchased from ESSEN Bioscience (Welwyn Garden City, UK).

Techniques: Incubation, Expressing, Fluorescence, Imaging

Journal: Biochemical Pharmacology

Article Title: A novel real time imaging platform to quantify macrophage phagocytosis

doi: 10.1016/j.bcp.2016.07.011

Figure Lengend Snippet: Pre-treatment with LPS leads to reduced E. coli phagocytosis by macrophages. (A) and (B) BMDMs (5 × 10 4 ) were primed with a range of LPS doses for 16 h and cells were incubated for another 24 h at 37 °C in culture media, after which 200 μg/ml green E. coli bioparticles were incubated with the macrophages in the IncuCyte platform for another hour. Green object counts in vehicle-treated macrophages were set to 100% and object counts in LPS pre-treated macrophage groups within the same experiment were normalized to demonstrate% reduction compared to the control. (C) Representative images from one experiment at one hour. Data from 5 biological replicates in technical duplicates are presented as mean ± SEM. Statistical analysis was carried out with Kruskal-Wallis and Dunn’s multiple comparisons post hoc test. Scale bar is 100 μm.

Article Snippet: Sterile IncuCyte™ green pHrodo® E. coli and Zymosan were purchased from ESSEN Bioscience (Welwyn Garden City, UK).

Techniques: Incubation

Journal: Biochemical Pharmacology

Article Title: A novel real time imaging platform to quantify macrophage phagocytosis

doi: 10.1016/j.bcp.2016.07.011

Figure Lengend Snippet: Comparison of the IncuCyte ZOOM® platform with other commonly used techniques for phagocytosis quantification.

Article Snippet: Sterile IncuCyte™ green pHrodo® E. coli and Zymosan were purchased from ESSEN Bioscience (Welwyn Garden City, UK).

Techniques: Comparison, Confocal Microscopy, Flow Cytometry, Imaging

Journal: Antibiotics

Article Title: Polymerase Chain Reaction-Lateral Flow Strip for Detecting Escherichia coli and Salmonella enterica Harboring bla CTX-M

doi: 10.3390/antibiotics14080745

Figure Lengend Snippet: Duplex PCR-LFS ( A ) and duplex PCR–gel electrophoresis ( B ) detection of E. coli with and without bla CTX-M , and non- E. coli with and without bla CTX-M . ( A ) Top line = control line; middle line = E. coli ; bottom line = bla CTX-M . E. coli carrying bla CTX-M shows two bands (species-specific and bla CTX-M ) (first strip from left). E. coli lacking bla CTX-M shows only the E. coli line (second strip). Non- E. coli strains carrying bla CTX-M show only the bla CTX-M line (third strip). Non- E. coli strains without bla CTX-M show no target lines (strips 4–8). Negative control = Klebsiella pneumoniae ATCC 13883. ( B ) Lane M = 100 bp DNA ladder; Lane 1 = E. coli with bla CTX-M ; Lane 2 = E. coli without bla CTX-M ; Lane 3 = non- E. coli with bla CTX-M ; Lanes 4–8 = non- E. coli without bla CTX-M ; Lane 9 = negative control ( Klebsiella pneumoniae ATCC 13883).

Article Snippet: Pure colonies of E. coli ATCC BAA-3303 and S. enterica strain SW060-1 were suspended in 1 mL of sterile saline, vortexed for 30 s, and adjusted to a turbidity of 1.00 using a turbidimeter.

Techniques: Nucleic Acid Electrophoresis, Control, Stripping Membranes, Negative Control

Journal: Antibiotics

Article Title: Polymerase Chain Reaction-Lateral Flow Strip for Detecting Escherichia coli and Salmonella enterica Harboring bla CTX-M

doi: 10.3390/antibiotics14080745

Figure Lengend Snippet: ROC curve showing triplicate standard deviation and theoretical sensitivity of duplex PCR-LFS detecting E. coli and Salmonella carrying bla CTX-M .

Article Snippet: Pure colonies of E. coli ATCC BAA-3303 and S. enterica strain SW060-1 were suspended in 1 mL of sterile saline, vortexed for 30 s, and adjusted to a turbidity of 1.00 using a turbidimeter.

Techniques: Standard Deviation

Journal: Antibiotics

Article Title: Polymerase Chain Reaction-Lateral Flow Strip for Detecting Escherichia coli and Salmonella enterica Harboring bla CTX-M

doi: 10.3390/antibiotics14080745

Figure Lengend Snippet: Detection limit of duplex PCR–gel electrophoresis ( A , C ) and duplex PCR-LFS ( B , D ) detecting E. coli ( A , B ) and Salmonella spp. ( C , D ) harboring bla CTX-M , spiked in raw pork mince. Dilutions: 2.5 × 10 5 to 2.5 CFU/mL for E. coli , and 4 × 10 5 to 4 CFU/mL for Salmonella . M = 100 bp DNA ladder; B = blank control (uninoculated).

Article Snippet: Pure colonies of E. coli ATCC BAA-3303 and S. enterica strain SW060-1 were suspended in 1 mL of sterile saline, vortexed for 30 s, and adjusted to a turbidity of 1.00 using a turbidimeter.

Techniques: Nucleic Acid Electrophoresis, Control

Journal: PLoS ONE

Article Title: Gene Expression Profiling during Early Acute Febrile Stage of Dengue Infection Can Predict the Disease Outcome

doi: 10.1371/journal.pone.0007892

Figure Lengend Snippet: List of genes differentially expressed in DF patients in relation to DHF patients based on the lowest p and highest fold change values.

Article Snippet: Second strand synthesis was performed by using E. coli DNA polymerase I and the double-stranded cDNA was used for in vitro transcription (IVT) for cRNA amplification by using Megascript T7 kit (Ambion).

Techniques: Ubiquitin Proteomics, Binding Assay, Membrane, Immunopeptidomics, Functional Assay, Sterility, Transformation Assay, Marker

Journal: Iranian Journal of Microbiology

Article Title: BCIG-SMAC medium and PMA-qPCR for differential detection of viable Escherichia coli in potable water

doi: 10.18502/ijm.v13i5.7427

Figure Lengend Snippet: Differentiation of viable cells in viable and dead cell mixtures by PMA-qPCR. Amplification curves for PMA-treated viable, dead, and a mixture of viable and dead cells of E. coli O157:H7 ATCC 43895 (A) and E. coli MTCC 3221 (B). The Cq values of DNA from viable cells of E. coli O157:H7 ATCC 43895 and E. coli MTCC 3221 has been compared with the Cq values of dead and mixture of viable and dead cells to indicate that DNA amplification from dead cells is completely inhibited as a result of the PMA-treatment (C). Each bar represents the average Cq value of a triplicate experiment ± standard deviation. ΔRn, fluorescence intensity change.

Article Snippet: Post-incubation, four aliquots comprising 400 μL each of E. coli O157:H7 ATCC 43895 and E. coli MTCC 3221 were spiked into autoclaved environmental water samples and filtered through sterile PES membrane filters ( ).

Techniques: Amplification, DNA Amplification, Standard Deviation, Fluorescence

Journal: Iranian Journal of Microbiology

Article Title: BCIG-SMAC medium and PMA-qPCR for differential detection of viable Escherichia coli in potable water

doi: 10.18502/ijm.v13i5.7427

Figure Lengend Snippet: PMA-qPCR quantification of pure cultures of E. coli O157:H7 ATCC 43895 and E. coli MTCC 3221. A series of dilutions of purified DNA (56 pg-0.007pg) extracted from PMA treated viable E. coli cells. The PMAqPCR could detect as low as 1 CFU/mL of E. coli.

Article Snippet: Post-incubation, four aliquots comprising 400 μL each of E. coli O157:H7 ATCC 43895 and E. coli MTCC 3221 were spiked into autoclaved environmental water samples and filtered through sterile PES membrane filters ( ).

Techniques: Purification

Journal: Iranian Journal of Microbiology

Article Title: BCIG-SMAC medium and PMA-qPCR for differential detection of viable Escherichia coli in potable water

doi: 10.18502/ijm.v13i5.7427

Figure Lengend Snippet: Box plot for PMA-untreated, PMA-treated qPCR and BCIG-SMAC counts of total viable E. coli (A) and E. coli O157:H7 (B) cells from the environmental water samples. Results are reported for n=71.

Article Snippet: Post-incubation, four aliquots comprising 400 μL each of E. coli O157:H7 ATCC 43895 and E. coli MTCC 3221 were spiked into autoclaved environmental water samples and filtered through sterile PES membrane filters ( ).

Techniques: