Xanthine (X8030) was purchased from Solarbio Life Science, Beijing, China. Middlebrook 7H9 Broth (271310) was purchased from BD/Difco, Franklin Lakes, NJ, United States. Amikacin (A602232), kanamycin (A100408), gentamicin (A100304), streptomycin (A100382), chloramphenicol (A100230), ciprofloxacin (A600310), rifampicin (A600812), and isoniazid (A600544) were purchased from Sangon Biotech, Shanghai, China. The Anti-His antibody (#9991) was purchased from Cell Signaling Technology, Danvers, MA, United States.
Amikacin
Manufactured by Sangon
Sourced in China
Amikacin is a semi-synthetic aminoglycoside antibiotic used to treat a variety of bacterial infections. It functions by inhibiting bacterial protein synthesis, leading to cell death.
Automatically generated - may contain errors
Lab products found in correlation
Gentamicin, by Sangon (6 mentions)
Kanamycin, by Sangon (3 mentions)
Ciprofloxacin, by Sangon (2 mentions)
Anti his antibody, by Cell Signaling Technology (1 mentions)
Rifampicin, by Sangon (1 mentions)
Streptomycin, by Sangon (1 mentions)
Chloramphenicol, by Sangon (1 mentions)
Microplate reader, by Bio-Rad (1 mentions)
Meropenem, by Yuanye Bio-Technology (1 mentions)
Cefotaxime, by Sangon (1 mentions)
Sodium pyruvate, by Merck Group (1 mentions)
Phenol, by Merck Group (1 mentions)
Flow cytometry reagent, by BD (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Yeast extract, by Thermo Fisher Scientific (1 mentions)
Ampicillin, by Merck Group (1 mentions)
Carboxy h2dcfda, by Thermo Fisher Scientific (1 mentions)
Kanamycin, by Merck Group (1 mentions)
Tryptone, by Thermo Fisher Scientific (1 mentions)
Chloramphenicol, by Merck Group (1 mentions)
7 protocols using amikacin
1
Xanthine-based Compound Screening
2
Bacterial Strains and Antimicrobial Cultivation
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Bacterial strains used in this study were from the collection of our laboratory and listed in Table S1 in the supplemental material. The antimicrobial agents amikacin, gentamicin, and kanamycin were purchased from a commercial source (Shanghai Sangon Biological Engineering Technology & Services Co. Ltd., China). A single colony was propagated in 3% NaCl Luria-Bertani (LB) (tryptone [10 g/liter], yeast extract [5 g/liter], NaCl [30 g/liter]) broth for 8 h at 30°C. The cultures were diluted to 1:100 using fresh 3% NaCl LB medium and grown at 30°C.
3
Assessing Sub-MICs of Antibiotics
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Antibiotic sub-MICs were selected by creating a growth curve. K. pneumoniae SW1780 was grown in the presence of 0 × MIC, 1/2 × MIC, 1/4 × MIC, 1/8 × MIC, 1/16 × MIC, 1/ 32 × MIC, 1/64 × MIC, 1/128 × MIC, 1/256 × MIC, 1/512 × MIC, 1/1024 × MIC, 1/2048 × MIC of meropenem (YuanYeBio-Technology, Shanghai, China), ciprofloxacin (Sango Biotech, Shanghai, China), cefotaxime (Sango Biotech, Shanghai, China) and amikacin (Sango Biotech, Shanghai, China). Samples were collected at one-hour intervals and the optical density was read at 595 nm (OD595) using a microplate reader (Bio-Rad, United States). The logarithmic growth rate of the antibiotic stress group and the non-antibiotic group was calculated by applying the logarithmic growth rate formula: U = [ln(Nt) -ln(N0)]/(t-t0), where Nt is the logarithmic growth late OD595 value, N0 is the logarithmic growth early OD595 value, and t-t0 is the logarithmic period time. One-way ANOVA was used to determine statistical differences. The highest antibiotic concentration that did not affect the growth of strain SW1780 was used as sub-MIC.
4
E. coli Growth and Genetic Manipulation
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E. coli K-12 strains (SI Appendix, Table S1 ) were grown aerobically at 37 °C in LB medium (63 (link)) with shaking at 200 rpm. Colony formation was on LB agar at 37 °C. Bacteriophage P1-mediated transduction (64 ) or CRISPR-based allelic exchange (24 (link)) was used for strain construction. Flow cytometry reagents were purchased from Becton Dickinson. Tryptone, yeast extract, powder for LB broth and agar, and carboxy-H2DCFDA were obtained from Thermo Fisher Scientific. Other reagents, including antimicrobials (ciprofloxacin, oxolinic acid, kanamycin, ampicillin, tetracycline, moxifloxacin, and chloramphenicol), phenol, dimethyl 2-oxoglutarate, cyclic AMP, sodium pyruvate, and DMSO were purchased from Sigma-Aldrich. Gentamicin, amikacin, hydrogen peroxide, chlorhexidine, ethanol, isopropanol, 1-butanol, potassium dichromate, sodium hypochlorite solution (5.2%), hydrogen chloride, and sodium hydroxide were purchased from Sangon Biotech. Meropenem (Shenghuaxi Pharmaceutical) and ceftriaxone (Roche) were from Zhongshan Hospital (Xiamen, China) pharmacy.
5
Antimicrobial Compound Evaluation Protocol
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Vancomycin (VAN), gentamicin, ciprofloxacin (CIP), linezolid, and amikacin were purchased from Sangon Biotech (Shanghai, China). Isopropoxy benzene guanidine (IBG) (batch number 20150506, content 99.9%) was synthesized by Guangzhou Insighter Biotechnology (Guangzhou, China). Dimethyl Sulphoxide (DMSO) (Dmreagent, Tianjing, China) was utilized as solvent to dissolve IBG. Anti-infective detergent benzalkonium chloride (BAC) was purchased from Aladdin Industrial Corporation (Shanghai, China). SYTOX® green nucleic acid stain agents and 3,3′-diethyloxacarbocyanine iodide [DiOC2(3)] (Thermo Fisher Scientific, Germany) were used as molecular probes.
6
Antibiotic Susceptibility Testing for A. baumannii
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The minimum inhibitory concentrations (MICs) of amikacin and gentamicin (Sangon Biotech Co., Ltd., Shanghai, China) for A. baumannii were determined on Mueller-Hinton agar plates by agar dilution according to the protocol recommended by the Clinical and Laboratory Standards Institute (39 ). MICs of meropenem and imipenem (Sangon Biotech Co., Ltd.) were tested in high-level aminoglycoside-resistant isolates. The results were interpreted according to the CLSI guidelines. Escherichia coli [American Type Culture Collection (ATCC) 25922] and A. baumannii (ATCC 19606) (ATCC, Manassas, VA, USA) were used as quality control strains.
7
Antibiotic Resistance Profiling Protocol
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Luria‐Bertani (LB) was purchased from Huankai Biotech Limited. All antibiotics tested, including tetracyclines (tetracycline), aminoglycosides (gentamicin, kanamycin and amikacin), penecillins (penecillin), cephalosporins (ceftazidime), quinolones (ofloxacin), macrolides (erythromycin), and polypeptide (polymyxin E) were purchased from Sangon Biotech Limited. Metabolome sample preparation and derivatization reagents including methanol, pyridine and methoxyamine hydrochloride were purchased from Thermo Fisher Scientific (American).
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