e coli attc 35218 standard strains  (ATCC)

Journal: International Journal of Molecular Sciences

Article Title: Design of New Antibacterial Enhancers Based on AcrB’s Structure and the Evaluation of Their Antibacterial Enhancement Activity

doi: 10.3390/ijms17111934

Figure Lengend Snippet: Dynamic growth curves of E. coli ATCC 35218. E. coli ATCC 35218 from the exponential phase was diluted with LB broth to 1.0 × 10 6 cfu/mL. Then, 64 mg/L artesunate (AS), DHA7 and DHA27 alone, and in combination with ampicillin (AMP)/tazobactam (TZB) were added to bacterial suspensions. Broth and 5% DMSO were negative control and solvent control, respectively. The bacterial growth was determined by measuring OD 600 at regular intervals. ( A ) The effect of drugs alone on growth of E. coli ATCC 35218; ( B ) The effect of drugs in combination of AMP on growth of E. coli ATCC 35218.

Article Snippet: The activities of these compounds were investigated using standard strain E. coli ATCC 35218.

Techniques: Negative Control, Solvent, Control

Journal: International Journal of Molecular Sciences

Article Title: Design of New Antibacterial Enhancers Based on AcrB’s Structure and the Evaluation of Their Antibacterial Enhancement Activity

doi: 10.3390/ijms17111934

Figure Lengend Snippet: Effect of DHA27 on antibacterial activity of AMP against 40 clinical E. coli strains.

Article Snippet: The activities of these compounds were investigated using standard strain E. coli ATCC 35218.

Techniques: Activity Assay, Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: Design of New Antibacterial Enhancers Based on AcrB’s Structure and the Evaluation of Their Antibacterial Enhancement Activity

doi: 10.3390/ijms17111934

Figure Lengend Snippet: Effect of DHA27 and PAβN on antibacterial effects of six antibiotics on six E. coli strains.

Article Snippet: The activities of these compounds were investigated using standard strain E. coli ATCC 35218.

Techniques: Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: Design of New Antibacterial Enhancers Based on AcrB’s Structure and the Evaluation of Their Antibacterial Enhancement Activity

doi: 10.3390/ijms17111934

Figure Lengend Snippet: Effect of DHA27 on daunorubicin accumulation within E. coli ATCC 35218. Daunorubicin accumulation within E. coli ATCC 35218 pretreated with artesunate (AS) and DHA27. ( A ) E. coli ATCC 35218 was treated with artesunate (AS 256, 512 mg/L) and DHA27 (32, 64, and 128 mg/L) and cultivated at 37 °C and 100 rpm for 6 h, and then harvested by centrifugation at 4000 rpm for 5 min. After washing three times and resuspending, the bacterial suspension was adjusted to OD 600 1.0. Bacteria were incubated with daunorubicin (40 mg/L) in the dark at 37 °C for 30 min. The numerical values on the column were increased percentage of treatment group than broth group; ( B ) E. coli ATCC 35218 was treated with artesunate (AS, 256 mg/L) and DHA27 (64 mg/L) and cultivated at 37 °C and 100 rpm for 6 h, and then harvested by centrifugation at 4000 rpm for 5 min. After washing three times and resuspending, the bacterial suspension was adjusted to OD 600 1.0. Bacteria were incubated with daunorubicin (40 mg/L) in the dark at 37 °C for 0, 10, 20, and 30 min. Then, 0.5 mL of bacteria was collected. Bacteria were washed three times and resuspended in PBS. Quantitative determination of daunorubicin accumulation in the presence of AS and DHA27 was determined by fluorospectrophotometry at the emission wavelength of 467 nm and excitation wavelength of 588 nm.* p < 0.05 compared with broth, ** p < 0.01 compared with broth.

Article Snippet: The activities of these compounds were investigated using standard strain E. coli ATCC 35218.

Techniques: Centrifugation, Suspension, Bacteria, Incubation

Journal: International Journal of Molecular Sciences

Article Title: Design of New Antibacterial Enhancers Based on AcrB’s Structure and the Evaluation of Their Antibacterial Enhancement Activity

doi: 10.3390/ijms17111934

Figure Lengend Snippet: Effect of DHA27 and PAβN in combination with different antibiotics on antibacterial activity against three E. coli strains.

Article Snippet: The activities of these compounds were investigated using standard strain E. coli ATCC 35218.

Techniques: Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: Design of New Antibacterial Enhancers Based on AcrB’s Structure and the Evaluation of Their Antibacterial Enhancement Activity

doi: 10.3390/ijms17111934

Figure Lengend Snippet: The effect of DHA27 on acrB mRNA expression. ( A ) The effect of DHA27 alone on the expression of acrB mRNA expression. E. coli clinical strains were treated with different concentration of DHA27 and inoculated into 50 mL of LB broth. Bacteria were cultivated at 37 °C in a heated and shaking environmental chamber of 6 h and then harvested by centrifugation at 1500 rpm for 5 min; ( B ) The AcrB mRNA expression of bacteria with FICIs > 0.5 was markedly lower than that of bacteria with FICIs < 0.5; ( C ) AcrB mRNA expression has negative correlation with antibacterial enhancement activities of DHA27 against them. E. coli clinical strains were inoculated into 50 mL of LB broth. Bacteria were cultivated at 37 °C in a heated and shaking environmental chamber to an OD 600 of 0.5 and then harvested by centrifugation at 1500 rpm for 5 min. RNA extraction and reverse transcription real-time PCR were performed. The real-time PCR products were detected using a MyiQ Color Fluorescence Real-Time Quantitative PCR kit (Bio-Rad). The gene fold changes were finally calculated according to the 2 −ΔΔ C t method for each transcript and expressed relative to the values from control group samples.

Article Snippet: The activities of these compounds were investigated using standard strain E. coli ATCC 35218.

Techniques: Expressing, Concentration Assay, Bacteria, Centrifugation, RNA Extraction, Reverse Transcription, Real-time Polymerase Chain Reaction, Fluorescence, Control

Journal: International Journal of Molecular Sciences

Article Title: Design of New Antibacterial Enhancers Based on AcrB’s Structure and the Evaluation of Their Antibacterial Enhancement Activity

doi: 10.3390/ijms17111934

Figure Lengend Snippet: Molecule docking of DHA27 binding to AcrB. The crystal structure of AcrB (PDB ID, 2DRD) was obtained from Protein Data Bank. The molecular docking results were analyzed and represented in the PyMOL 1.3 visualization software. Proteins are shown in orange. The compound is shown as thick sticks colored according to the atom type (red, oxygen; cyan, carbon; white, hydrogen; dark blue, nitrogen, khaki, sulfur), and residues are shown in bluish violet and marked. ( A ) Longitudinal section of substrate translocation pathway when DHA27 was docked with AcrB when PDB ID 2DRD was used; ( B ) Cartoon view of DHA27 docked with AcrB when PDB ID 2DRD was used; ( C ) Effect of DHA27 on binding polypeptides from AcrB and its mutant peptides. Daunorubicin accumulation within E. coli ATCC 35218 pretreated with DHA27 and polypeptides; 64 mg/L of DHA27 and polypeptide (molar ratio: 1:1) were cultured for 30 min at 37 °C. This mixture was added to the E. coli concentrations of 1.0 × 10 6 cfu/mL and cultured for 6 h at 37 °C in a heated and shaking environmental chamber. The bacterial pellet was resuspended, and the bacterial suspension was adjusted to an OD 600 of 1.0. After co-culture with daunorubicin (40 mg/L) in the dark at 37 °C for 30 min, the bacteria were centrifuged at 3000 rpm for 5 min to harvest the bacterial pellet. After washing with PBS (0.2 mM, pH 7.2) five times, fluorospectrophotometry was used to observe daunorubicin accumulation within E. coli . The emission wavelength was 467 nm, and the excitation wavelength was 588 nm. ** p < 0.01 compared with broth; ## p < 0.01 compared with DHA27.

Article Snippet: The activities of these compounds were investigated using standard strain E. coli ATCC 35218.

Techniques: Binding Assay, Software, Translocation Assay, Mutagenesis, Cell Culture, Suspension, Co-Culture Assay, Bacteria

Journal: International Journal of Molecular Sciences

Article Title: Design of New Antibacterial Enhancers Based on AcrB’s Structure and the Evaluation of Their Antibacterial Enhancement Activity

doi: 10.3390/ijms17111934

Figure Lengend Snippet: Dynamic growth curves of E. coli ATCC 35218. E. coli ATCC 35218 from the exponential phase was diluted with LB broth to 1.0 × 10 6 cfu/mL. Then, 64 mg/L artesunate (AS), DHA7 and DHA27 alone, and in combination with ampicillin (AMP)/tazobactam (TZB) were added to bacterial suspensions. Broth and 5% DMSO were negative control and solvent control, respectively. The bacterial growth was determined by measuring OD 600 at regular intervals. ( A ) The effect of drugs alone on growth of E. coli ATCC 35218; ( B ) The effect of drugs in combination of AMP on growth of E. coli ATCC 35218.

Article Snippet: The aforementioned results indicated that DHA27 had the antibacterial enhancement activity of AMP against not only E. coli standard strain ATCC 35218 but also most clinical strains.

Techniques: Negative Control, Solvent, Control

Journal: International Journal of Molecular Sciences

Article Title: Design of New Antibacterial Enhancers Based on AcrB’s Structure and the Evaluation of Their Antibacterial Enhancement Activity

doi: 10.3390/ijms17111934

Figure Lengend Snippet: Effect of DHA27 on antibacterial activity of AMP against 40 clinical E. coli strains.

Article Snippet: The aforementioned results indicated that DHA27 had the antibacterial enhancement activity of AMP against not only E. coli standard strain ATCC 35218 but also most clinical strains.

Techniques: Activity Assay, Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: Design of New Antibacterial Enhancers Based on AcrB’s Structure and the Evaluation of Their Antibacterial Enhancement Activity

doi: 10.3390/ijms17111934

Figure Lengend Snippet: Effect of DHA27 and PAβN on antibacterial effects of six antibiotics on six E. coli strains.

Article Snippet: The aforementioned results indicated that DHA27 had the antibacterial enhancement activity of AMP against not only E. coli standard strain ATCC 35218 but also most clinical strains.

Techniques: Concentration Assay

Journal: International Journal of Molecular Sciences

Article Title: Design of New Antibacterial Enhancers Based on AcrB’s Structure and the Evaluation of Their Antibacterial Enhancement Activity

doi: 10.3390/ijms17111934

Figure Lengend Snippet: Effect of DHA27 on daunorubicin accumulation within E. coli ATCC 35218. Daunorubicin accumulation within E. coli ATCC 35218 pretreated with artesunate (AS) and DHA27. ( A ) E. coli ATCC 35218 was treated with artesunate (AS 256, 512 mg/L) and DHA27 (32, 64, and 128 mg/L) and cultivated at 37 °C and 100 rpm for 6 h, and then harvested by centrifugation at 4000 rpm for 5 min. After washing three times and resuspending, the bacterial suspension was adjusted to OD 600 1.0. Bacteria were incubated with daunorubicin (40 mg/L) in the dark at 37 °C for 30 min. The numerical values on the column were increased percentage of treatment group than broth group; ( B ) E. coli ATCC 35218 was treated with artesunate (AS, 256 mg/L) and DHA27 (64 mg/L) and cultivated at 37 °C and 100 rpm for 6 h, and then harvested by centrifugation at 4000 rpm for 5 min. After washing three times and resuspending, the bacterial suspension was adjusted to OD 600 1.0. Bacteria were incubated with daunorubicin (40 mg/L) in the dark at 37 °C for 0, 10, 20, and 30 min. Then, 0.5 mL of bacteria was collected. Bacteria were washed three times and resuspended in PBS. Quantitative determination of daunorubicin accumulation in the presence of AS and DHA27 was determined by fluorospectrophotometry at the emission wavelength of 467 nm and excitation wavelength of 588 nm.* p < 0.05 compared with broth, ** p < 0.01 compared with broth.

Article Snippet: The aforementioned results indicated that DHA27 had the antibacterial enhancement activity of AMP against not only E. coli standard strain ATCC 35218 but also most clinical strains.

Techniques: Centrifugation, Suspension, Bacteria, Incubation

Journal: International Journal of Molecular Sciences

Article Title: Design of New Antibacterial Enhancers Based on AcrB’s Structure and the Evaluation of Their Antibacterial Enhancement Activity

doi: 10.3390/ijms17111934

Figure Lengend Snippet: Effect of DHA27 and PAβN in combination with different antibiotics on antibacterial activity against three E. coli strains.

Article Snippet: The aforementioned results indicated that DHA27 had the antibacterial enhancement activity of AMP against not only E. coli standard strain ATCC 35218 but also most clinical strains.

Techniques: Activity Assay

Journal: International Journal of Molecular Sciences

Article Title: Design of New Antibacterial Enhancers Based on AcrB’s Structure and the Evaluation of Their Antibacterial Enhancement Activity

doi: 10.3390/ijms17111934

Figure Lengend Snippet: The effect of DHA27 on acrB mRNA expression. ( A ) The effect of DHA27 alone on the expression of acrB mRNA expression. E. coli clinical strains were treated with different concentration of DHA27 and inoculated into 50 mL of LB broth. Bacteria were cultivated at 37 °C in a heated and shaking environmental chamber of 6 h and then harvested by centrifugation at 1500 rpm for 5 min; ( B ) The AcrB mRNA expression of bacteria with FICIs > 0.5 was markedly lower than that of bacteria with FICIs < 0.5; ( C ) AcrB mRNA expression has negative correlation with antibacterial enhancement activities of DHA27 against them. E. coli clinical strains were inoculated into 50 mL of LB broth. Bacteria were cultivated at 37 °C in a heated and shaking environmental chamber to an OD 600 of 0.5 and then harvested by centrifugation at 1500 rpm for 5 min. RNA extraction and reverse transcription real-time PCR were performed. The real-time PCR products were detected using a MyiQ Color Fluorescence Real-Time Quantitative PCR kit (Bio-Rad). The gene fold changes were finally calculated according to the 2 −ΔΔ C t method for each transcript and expressed relative to the values from control group samples.

Article Snippet: The aforementioned results indicated that DHA27 had the antibacterial enhancement activity of AMP against not only E. coli standard strain ATCC 35218 but also most clinical strains.

Techniques: Expressing, Concentration Assay, Bacteria, Centrifugation, RNA Extraction, Reverse Transcription, Real-time Polymerase Chain Reaction, Fluorescence, Control

Journal: International Journal of Molecular Sciences

Article Title: Design of New Antibacterial Enhancers Based on AcrB’s Structure and the Evaluation of Their Antibacterial Enhancement Activity

doi: 10.3390/ijms17111934

Figure Lengend Snippet: Molecule docking of DHA27 binding to AcrB. The crystal structure of AcrB (PDB ID, 2DRD) was obtained from Protein Data Bank. The molecular docking results were analyzed and represented in the PyMOL 1.3 visualization software. Proteins are shown in orange. The compound is shown as thick sticks colored according to the atom type (red, oxygen; cyan, carbon; white, hydrogen; dark blue, nitrogen, khaki, sulfur), and residues are shown in bluish violet and marked. ( A ) Longitudinal section of substrate translocation pathway when DHA27 was docked with AcrB when PDB ID 2DRD was used; ( B ) Cartoon view of DHA27 docked with AcrB when PDB ID 2DRD was used; ( C ) Effect of DHA27 on binding polypeptides from AcrB and its mutant peptides. Daunorubicin accumulation within E. coli ATCC 35218 pretreated with DHA27 and polypeptides; 64 mg/L of DHA27 and polypeptide (molar ratio: 1:1) were cultured for 30 min at 37 °C. This mixture was added to the E. coli concentrations of 1.0 × 10 6 cfu/mL and cultured for 6 h at 37 °C in a heated and shaking environmental chamber. The bacterial pellet was resuspended, and the bacterial suspension was adjusted to an OD 600 of 1.0. After co-culture with daunorubicin (40 mg/L) in the dark at 37 °C for 30 min, the bacteria were centrifuged at 3000 rpm for 5 min to harvest the bacterial pellet. After washing with PBS (0.2 mM, pH 7.2) five times, fluorospectrophotometry was used to observe daunorubicin accumulation within E. coli . The emission wavelength was 467 nm, and the excitation wavelength was 588 nm. ** p < 0.01 compared with broth; ## p < 0.01 compared with DHA27.

Article Snippet: The aforementioned results indicated that DHA27 had the antibacterial enhancement activity of AMP against not only E. coli standard strain ATCC 35218 but also most clinical strains.

Techniques: Binding Assay, Software, Translocation Assay, Mutagenesis, Cell Culture, Suspension, Co-Culture Assay, Bacteria