Amikacin disulfate salt

Aliquots of bacterial cultures grown as described above were taken at time points indicated in the text, washed twice in sterile phosphate-buffered saline, diluted between 10⁻⁵ to 10⁻⁶, and 100 μL spread on either LB agar plates with or without specified antibiotics (2 μg/mL amikacin disulfate salt or 7 μg/mL nitrofurantoin [both Sigma-Aldrich]) or 50% (vol/vol) healthy female human urine containing 1.5% (wt/vol) agar. Plates were incubated at 33°C in a standard office scanner (Epson Perfection V370 photo scanner, J232D) placed in an incubator, and images were taken every 20 min over a 48-h period. Analysis of appearance time and apparent growth rate of colonies was adapted from Levin-Reisman et al. (18 (link)) using a modified code by Miles Priestman available at https://github.com/mountainpenguin/NQBMatlab. At least two independent experiments were conducted for each ScanLag experiment shown in the figures.

K. pneumoniae and E. coli strains were purchased from the American Type Culture Collection (Manassas, VA) or the National Collection of Type Cultures (London, United Kingdom). Cation-adjusted Mueller-Hinton broth was used for antibacterial susceptibility testing and time-kill assays, and heart infusion agar was used for the enumeration of viable cells.
TP0586532 was synthesized by the Department of Medicinal Chemistry, Taisho Pharmaceutical Co., Ltd. (Saitama, Japan). Amikacin disulfate salt, colistin sulfate, piperacillin, tigecycline hydrate, and azithromycin dihydrate were purchased from Sigma-Aldrich (St. Louis, MO). Cefepime hydrochloride was purchased from US Pharmacopeia (Rockville, MD). Meropenem trihydrate were purchased from US Pharmacopeia or FUJIFILM Wako Pure Chemical Corporation (Osaka, Japan). Ciprofloxacin hydrochloride monohydrate was purchased from FUJIFILM Wako Pure Chemical Corporation.

K. pneumoniae and E. coli strains were purchased from the American Type Culture Collection (Manassas, VA) or the National Collection of Type Cultures (London, United Kingdom). Cation-adjusted Mueller-Hinton broth was used for antibacterial susceptibility testing and time-kill assays, and heart infusion agar was used for the enumeration of viable cells.
TP0586532 was synthesized by the Department of Medicinal Chemistry, Taisho Pharmaceutical Co., Ltd. (Saitama, Japan). Amikacin disulfate salt, colistin sulfate, piperacillin, tigecycline hydrate, and azithromycin dihydrate were purchased from Sigma-Aldrich (St. Louis, MO). Cefepime hydrochloride was purchased from US Pharmacopeia (Rockville, MD). Meropenem trihydrate were purchased from US Pharmacopeia or FUJIFILM Wako Pure Chemical Corporation (Osaka, Japan). Ciprofloxacin hydrochloride monohydrate was purchased from FUJIFILM Wako Pure Chemical Corporation.