mid logarithmic phase cultures  (tiangen biotech co)

Journal: AMB Express

Article Title: Repurposed bunamidine disrupts envelope energetics and induces ROS-mediated non-lytic killing

doi: 10.1186/s13568-026-02016-6

Figure Lengend Snippet: Antibacterial activity of BUN against MRSA. A Chemical structure of bunamidine hydrochloride (BUN). B MIC determinations of BUN against MRSA ATCC 43300 and USA300 in MH and TSB media. C XTT assays showing concentration-dependent inhibition of metabolic activity by BUN. D Comparison of OD 630 changes over time between BUN and the bacteriostatic comparator LZD. E Time-kill kinetics demonstrating concentration-dependent bactericidal activity of BUN. F Live/dead staining of MRSA using SYTO9/PI probes after 2 h exposure to BUN. G Quantification of PI-positive (dead) cells following BUN treatment. Data are presented as mean ± SD from at least three independent experiments. Statistical analyses were performed using one-way ANOVA with post hoc comparisons; ns, not significant; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001

Article Snippet: Mid-logarithmic-phase MRSA ATCC 43300 cultures were diluted in Mueller–Hinton (MH) broth to approximately 1 × 10 6 CFU/mL.

Techniques: Activity Assay, Concentration Assay, Inhibition, Comparison, Staining

Journal: AMB Express

Article Title: Repurposed bunamidine disrupts envelope energetics and induces ROS-mediated non-lytic killing

doi: 10.1186/s13568-026-02016-6

Figure Lengend Snippet: Antibiofilm and anti-persister activity of BUN against MRSA. A , B Biofilm inhibition ( A ) and eradication ( B ) of MRSA ATCC 43300 determined by CV (biomass) and XTT (metabolic activity) assays after exposure to BUN (0–16 µg/mL) in TSBg for 24 h. Statistical significance was evaluated by one-way ANOVA with Dunnett’s test versus control (ns, * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001). C , D CLSM images of biofilms stained with SYTO9 (green) and PI (red) under inhibition ( C ) and eradication ( D ) conditions following BUN treatment (4 or 8 µg/mL). Scale bars, 20 µm. E , F Quantification of live/dead cell ratios from representative CLSM fields. G Time-kill curves of biofilm-associated persister cells from MRSA ATCC 43300 and USA300 challenged with BUN (4 µg/mL) or comparators [VAN and DAP, 10 × MIC]. Dashed line indicates the limit of detection. Data are presented as mean ± SD from independent experiments

Article Snippet: Mid-logarithmic-phase MRSA ATCC 43300 cultures were diluted in Mueller–Hinton (MH) broth to approximately 1 × 10 6 CFU/mL.

Techniques: Activity Assay, Inhibition, Control, Staining

Journal: AMB Express

Article Title: Repurposed bunamidine disrupts envelope energetics and induces ROS-mediated non-lytic killing

doi: 10.1186/s13568-026-02016-6

Figure Lengend Snippet: Structural perturbation, hemolytic assessment, and ROS involvement in BUN activity. A Transmission electron microscopy images of MRSA ATCC 43300 showing morphological changes after BUN treatment compared with untreated controls. B Hemolysis of freshly isolated human red blood cells in the presence of BUN. Hemoglobin release was measured spectrophotometrically. C Effect of glutathione (GSH) supplementation on the MIC of BUN against MRSA ATCC 43300 and USA300. D Confocal fluorescence imaging of intracellular ROS in MRSA ATCC 43300 using DCFH-DA probe, with or without GSH supplementation. Bright-field images are shown for comparison. Scale: 200 μm

Article Snippet: Mid-logarithmic-phase MRSA ATCC 43300 cultures were diluted in Mueller–Hinton (MH) broth to approximately 1 × 10 6 CFU/mL.

Techniques: Activity Assay, Transmission Assay, Electron Microscopy, Isolation, Fluorescence, Imaging, Comparison

Journal: ACS Omega

Article Title: High-Throughput Screening Identifies Synthetic Peptides with Antibacterial Activity against Mycobacterium abscessus and Serum Stability

doi: 10.1021/acsomega.2c02844

Figure Lengend Snippet: (A) Representative colonies of Mabs ATCC 19977S and Mabs ATCC 19977R morphotypes. (B) CTO-labeled (left) or AF647-labeled (right) Mabs ATCC 19977S binding to a peptide spot on the microarray. (C) Statistics of bacteria-binding signals to the peptide microarrays. (D) Workflow to select and test Mabs peptides.

Article Snippet: Mabs ATCC 19977S and ATCC 19977R mid-logarithmic-phase cultures (OD 600 0.5–0.6) were centrifuged (3715 g ) for 2 min, washed in prewarmed M7H9, resuspended in M7H9, MHB, or CAMHB, and diluted to 10 6 CFU/mL.

Techniques: Labeling, Binding Assay, Microarray

Journal: ACS Omega

Article Title: High-Throughput Screening Identifies Synthetic Peptides with Antibacterial Activity against Mycobacterium abscessus and Serum Stability

doi: 10.1021/acsomega.2c02844

Figure Lengend Snippet: Growth inhibition assays with the 27 peptides that interacted with Mabs during the peptide microarray screening. MIC assays with Mabs 19977S and Mabs 19977R were performed in (A,B) M7H9, (C,D) CAMHB, or (E,F) MHB with peptide (100 μM) and EDTA (100 μM) or peptide (100 μM) alone. All experiments were incubated at 37 °C for 72 h. Peptides that reduced Mabs growth by ≥50% (red hatched lines), when compared to the growth control OD 600 , were considered active. Panels highlight six peptides with consistent inhibitory activity against Mabs 19977S and Mabs 19977R in different media. ASU2001—green circles; ASU2009—blue circles; ASU2019—purple circles; ASU2056—orange circles; ASU2059—olive green circles; and ASU2060—red circles. All experiments were performed in triplicate with the average of all three replicates plotted for each peptide.

Article Snippet: Mabs ATCC 19977S and ATCC 19977R mid-logarithmic-phase cultures (OD 600 0.5–0.6) were centrifuged (3715 g ) for 2 min, washed in prewarmed M7H9, resuspended in M7H9, MHB, or CAMHB, and diluted to 10 6 CFU/mL.

Techniques: Inhibition, Peptide Microarray, Incubation, Activity Assay

Journal: ACS Omega

Article Title: High-Throughput Screening Identifies Synthetic Peptides with Antibacterial Activity against Mycobacterium abscessus and Serum Stability

doi: 10.1021/acsomega.2c02844

Figure Lengend Snippet: IC 50 and MIC values of (A) ASU2001, (B) ASU2009, (C) ASU2019, (D) ASU2056, (E) ASU2059, and (F) ASU2060 peptides against Mabs ATCC 19977R rough morphotype in MHB supplemented with EDTA (100 μM) for 96 h at 37 °C. Three independent experiments were performed with the average of all three replicates plotted for each peptide. Absorbance (OD 600 ) values were normalized to the growth control (100%), and IC 50 values were determined by nonlinear regression.

Article Snippet: Mabs ATCC 19977S and ATCC 19977R mid-logarithmic-phase cultures (OD 600 0.5–0.6) were centrifuged (3715 g ) for 2 min, washed in prewarmed M7H9, resuspended in M7H9, MHB, or CAMHB, and diluted to 10 6 CFU/mL.

Techniques:

Journal: ACS Omega

Article Title: High-Throughput Screening Identifies Synthetic Peptides with Antibacterial Activity against Mycobacterium abscessus and Serum Stability

doi: 10.1021/acsomega.2c02844

Figure Lengend Snippet: (A) Representative colonies of Mabs ATCC 19977S and Mabs ATCC 19977R morphotypes. (B) CTO-labeled (left) or AF647-labeled (right) Mabs ATCC 19977S binding to a peptide spot on the microarray. (C) Statistics of bacteria-binding signals to the peptide microarrays. (D) Workflow to select and test Mabs peptides.

Article Snippet: Mabs ATCC 19977S and ATCC 19977R mid-logarithmic-phase cultures (OD 600 0.5–0.6) were centrifuged (3715 g ) for 2 min, washed in prewarmed M7H9, resuspended in M7H9, MHB, or CAMHB, and diluted to 10 6 CFU/mL.

Techniques: Labeling, Binding Assay, Microarray, Bacteria

Journal: ACS Omega

Article Title: High-Throughput Screening Identifies Synthetic Peptides with Antibacterial Activity against Mycobacterium abscessus and Serum Stability

doi: 10.1021/acsomega.2c02844

Figure Lengend Snippet: Growth inhibition assays with the 27 peptides that interacted with Mabs during the peptide microarray screening. MIC assays with Mabs 19977S and Mabs 19977R were performed in (A,B) M7H9, (C,D) CAMHB, or (E,F) MHB with peptide (100 μM) and EDTA (100 μM) or peptide (100 μM) alone. All experiments were incubated at 37 °C for 72 h. Peptides that reduced Mabs growth by ≥50% (red hatched lines), when compared to the growth control OD 600 , were considered active. Panels highlight six peptides with consistent inhibitory activity against Mabs 19977S and Mabs 19977R in different media. ASU2001—green circles; ASU2009—blue circles; ASU2019—purple circles; ASU2056—orange circles; ASU2059—olive green circles; and ASU2060—red circles. All experiments were performed in triplicate with the average of all three replicates plotted for each peptide.

Article Snippet: Mabs ATCC 19977S and ATCC 19977R mid-logarithmic-phase cultures (OD 600 0.5–0.6) were centrifuged (3715 g ) for 2 min, washed in prewarmed M7H9, resuspended in M7H9, MHB, or CAMHB, and diluted to 10 6 CFU/mL.

Techniques: Inhibition, Peptide Microarray, Incubation, Control, Activity Assay

Journal: ACS Omega

Article Title: High-Throughput Screening Identifies Synthetic Peptides with Antibacterial Activity against Mycobacterium abscessus and Serum Stability

doi: 10.1021/acsomega.2c02844

Figure Lengend Snippet: IC 50 and MIC values of (A) ASU2001, (B) ASU2009, (C) ASU2019, (D) ASU2056, (E) ASU2059, and (F) ASU2060 peptides against Mabs ATCC 19977R rough morphotype in MHB supplemented with EDTA (100 μM) for 96 h at 37 °C. Three independent experiments were performed with the average of all three replicates plotted for each peptide. Absorbance (OD 600 ) values were normalized to the growth control (100%), and IC 50 values were determined by nonlinear regression.

Article Snippet: Mabs ATCC 19977S and ATCC 19977R mid-logarithmic-phase cultures (OD 600 0.5–0.6) were centrifuged (3715 g ) for 2 min, washed in prewarmed M7H9, resuspended in M7H9, MHB, or CAMHB, and diluted to 10 6 CFU/mL.

Techniques: Control