PA14, mvfR-, aphC-, ahpF-, trxB2-, ahpB- and dps- cells were grown at 37 °C 200 rpm in LB Lenox media until mid-exponential phase (OD600nm 2) then exposed to 300 mM hydrogen peroxide (H2O2) for 1 hour under the same incubating conditions. Before (t = 0) and after (t = 1 h) H2O2 addition, a 100 μL sample of each culture was collected, diluted and plated on LB agar plates to quantify the total number of bacteria (t = 0) and the surviving bacteria (t = 1 h). Colony forming units (CFUs) were counted after 24 h incubation at 37 °C. Tolerance to Meropenem was assessed the same way except the cells were grown in 1% TSB media and the killing was performed for 24 hours in presence of 10 μg/mL Meropenem (Sandoz, USA).
Meropenem
Manufactured by Sandoz
Sourced in United States
Meropenem is a laboratory instrument used for the preparation and analysis of pharmaceutical samples. It is a high-performance liquid chromatography (HPLC) device designed to accurately measure the concentration of the antibiotic meropenem in various solutions.
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Lab products found in correlation
Zidebactam, by MedKoo Biosciences (1 mentions)
Cefepime, by Merck Group (1 mentions)
Doxycycline, by Merck Group (1 mentions)
Vancomycin, by Merck Group (1 mentions)
Levofloxacin, by Merck Group (1 mentions)
Piperacillin tazobactam, by Merck Group (1 mentions)
Ampicillin, by Merck Group (1 mentions)
Penicillin, by Merck Group (1 mentions)
Streptomycin, by Merck Group (1 mentions)
Polymyxin b, by Merck Group (1 mentions)
Colistin, by Merck Group (1 mentions)
Imipenem, by Merck Group (1 mentions)
Camhb, by BD (1 mentions)
6 protocols using meropenem
1
Oxidative and Antibiotic Stress Assays
2
Antibiotic Susceptibility Testing Protocol
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Bacterial isolates were subjected to broth microdilution susceptibility testing, performed according to Clinical and Laboratory Standards Institute (CLSI) methods (46 ), using panels prepared in-house. Meropenem was purchased from Sandoz, and all other antibiotics were from Sigma-Aldrich. Xeruborbactam was synthesized at Qpex Biopharma, San Diego, CA. Avibactam was purchased from eNovation Chemicals LLC, Bridgewater, NJ, USA; zidebactam was from MedKoo Biosciences, Inc., Morrisville, NC, and nacubactam was from Advanced ChemBlocks, Hayward, CA. Durlobactam was synthesized at Acme Bioscience, Palo Alto, CA, USA.
3
Antibiotic Susceptibility of A. baumannii
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The MIC90 for colistin (Sigma, St. Louis, MO, USA, #C4461), polymyxin B (Sigma, #P0972), penicillin (Sigma, #5161), streptomycin (Sigma, #S9137), ampicillin (Sigma, #171254), vancomycin (Sigma, #SBR00001), doxycycline (Sigma, #D1822), cefepime (Sigma, #Y0000633), meropenem (Sandoz, Basel, Switzerland, #0781-3000-94), piperacillin/tazobactam (Sigma, #93129, #T2820), imipenem (Sigma, #I0090000), and levofloxacin (Sigma, #40922) was determined for wza# and wza-Rev. Briefly, A. baumannii strains were grown to OD600 of ~0.4, washed twice with PBS, resuspended at an OD600 of 0.4 (~1 × 108 CFU/mL), and diluted 1:100 in RPMI supplemented with 10% TSB. Antibiotics were serially diluted two-fold from 250 μg/mL to 1.95 μg/mL. Bacteria were added to the diluted antibiotics and grown for 20 h. MIC90 was determined as the concentration at which inhibited 90% or more growth relative to untreated.
4
Meropenem-Vaborbactam Combination Susceptibility
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Bacteria were cultured on Mueller-Hinton agar and tested on cation-adjusted Mueller-Hinton broth (CAMHB) (Becton Dickinson, Sparks, MD). Stock solutions of meropenem (Sandoz, Princeton, NJ) (10 to 50 mg/ml in distilled H2O) and vaborbactam (The Medicines Company, San Diego, CA) (5 to 10 mg/ml in 90% dimethyl sulfoxide) were stored at −80°C until use.
Broth microdilution susceptibility testing was performed according to Clinical and Laboratory Standards Institute methods (40 ), using panels prepared in-house. A checkerboard assay conforming to the Moody procedures described in the Clinical Microbiology Procedures Handbook (41 ) was used to evaluate the effect of various concentrations of vaborbactam on the meropenem MIC.
Broth microdilution susceptibility testing was performed according to Clinical and Laboratory Standards Institute methods (40 ), using panels prepared in-house. A checkerboard assay conforming to the Moody procedures described in the Clinical Microbiology Procedures Handbook (41 ) was used to evaluate the effect of various concentrations of vaborbactam on the meropenem MIC.
5
Broth Microdilution Susceptibility Testing Protocol
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The bacterial isolates were subjected to broth microdilution susceptibility testing, performed according to Clinical and Laboratory Standards Institute (CLSI) methods (26 ), using panels prepared in-house. Meropenem was purchased from Sandoz; all other antibiotics were from Sigma-Aldrich. QPC7728 and vaborbactam were synthesized at Qpex Biopharma, Inc., San Diego, CA. Avibactam was purchased from eNovation Chemicals LLC, Bridgewater, NJ, USA, and relebactam was synthesized at Acme Bioscience, Palo Alto, CA, USA, or purchased from AChemBlock, Burlingame, CA, USA.
6
Antibiotics Potentiation by Vaborbactam
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Bacterial isolates were subjected to broth microdilution susceptibility testing, performed according to Clinical and Laboratory Standards Institute (CLSI) methods (53 ), using panels prepared in-house. A checkerboard assay conforming to the procedures described by Moody in the Clinical Microbiology Procedures Handbook (54 ) was used to evaluate the effects of various concentrations of vaborbactam on the MICs of various antibiotics. MPC16 and MPCmax (where MPC stands for the m inimal p otentiating c oncentration) values were used to define the potency of vaborbactam. MPC16 was defined as the concentration of vaborbactam that was required to reduce the antibiotic MIC 16-fold. MPCmax was defined as the concentration of vaborbactam that achieved the maximal effect in increasing antibiotic potency.
Meropenem was purchased from Sandoz; all other antibiotics were from Sigma-Aldrich. Vaborbactam was synthesized at The Medicines Company, San Diego, CA (lot P-232-159-2).
Meropenem was purchased from Sandoz; all other antibiotics were from Sigma-Aldrich. Vaborbactam was synthesized at The Medicines Company, San Diego, CA (lot P-232-159-2).
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