4-(2-Hydroxyethyl)piperazine-1-ethanesulfonic acid (HEPES, purity > 99.5%, CAS#7365-45-9), potassium chloride (KCl, purity > 99.0%, CAS#7447-40-7), agarose (purity > 99.0%, CAS#: 9012-36-6), chloroform (purity > 99.0%, CAS: 67-66-3), isopropyl alcohol (purity > 99.0%, CAS#: 67-63-0), and alcohol (purity > 99.0%, CAS#: 64-17-5) was purchased from Sigma-Aldrich. RNase inhibitor (5 KU), pET-28b plasmid, and all the DNA were provided by Sangon Biotech. 1,2-Diphytanoyl-sn-glycero-3-phosphocholine (DPHPC) was purchased from Avanti. PrimeSTAR HS DNA polymerase was purchased from TaKaRa. Imipenem (CAS#: 64221-86-9) was purchased from MSD.
Imipenem
Manufactured by MSD
Sourced in Germany, United States
Imipenem is a broad-spectrum antibiotic used in the treatment of a variety of bacterial infections. It belongs to the carbapenem class of antibiotics and functions by inhibiting cell wall synthesis in bacteria, leading to cell death. Imipenem is often used in combination with other antibiotics to treat complex or multidrug-resistant infections.
Automatically generated - may contain errors
Lab products found in correlation
Ciprofloxacin, by Bayer (15 mentions)
Metronidazole, by Fresenius (14 mentions)
Ampicillin, by Ratiopharm (8 mentions)
Ampicillin plus sulbactam, by Ratiopharm (6 mentions)
Cefotaxime, by Merck Group (3 mentions)
Sulbactam, by Ratiopharm (3 mentions)
Tigecycline, by Pfizer (3 mentions)
Ciprofloxacin, by Merck Group (2 mentions)
Gentamicin, by Merck Group (2 mentions)
Kanamycin, by MP Biomedicals (2 mentions)
Thioglycolate enrichment broths, by Thermo Fisher Scientific (2 mentions)
Agarose, by Merck Group (1 mentions)
Primestar hs dna polymerase, by Takara Bio (1 mentions)
Chloroform, by Merck Group (1 mentions)
Isopropyl alcohol, by Merck Group (1 mentions)
Female balb c mice, by Orient Bio (1 mentions)
Ceftriaxone, by Merck Group (1 mentions)
Ofloxacin, by Merck Group (1 mentions)
Nitrofurantoin, by Merck Group (1 mentions)
Norfloxacin, by Merck Group (1 mentions)
25 protocols using imipenem
1
Protein Expression and Purification Protocols
2
Antibiotic Cocktail Induced Gut Dysbiosis
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Female BALB/c mice weighing 16-20 g (3 weeks old) were purchased from Orient Bio Inc. (Seongnam, Korea) and treated in accordance with the guidelines of the Institutional Animal Care and Use Committee of the Asan Medical Center and Ulsan University College of Medicine (identification code No. 2014-02-182).
For the induction of gut dysbiosis, mice were treated with an antibiotic cocktail in their drinking water for 2 weeks before primary sensitization with ampicillin (1 g/L; Ratiopharm, Ulm, Germany), vancomycin (500 mg/L; Cell Pharm, Hannover, Germany), ciprofloxacin (200 mg/L; Bayer Vital, Leverkusen, Germany), imipenem (250 mg/L; MSD, Haar, Germany), and metronidazole (1 g/L; Fresenius, Bad Homburg, Germany).25 (link)
For the induction of gut dysbiosis, mice were treated with an antibiotic cocktail in their drinking water for 2 weeks before primary sensitization with ampicillin (1 g/L; Ratiopharm, Ulm, Germany), vancomycin (500 mg/L; Cell Pharm, Hannover, Germany), ciprofloxacin (200 mg/L; Bayer Vital, Leverkusen, Germany), imipenem (250 mg/L; MSD, Haar, Germany), and metronidazole (1 g/L; Fresenius, Bad Homburg, Germany).25 (link)
3
Generation of Abiotic Mice for Stable C. jejuni Colonization
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IL-10−/− mice (female and male, all in C57BL/6j background) were reared under specific pathogen free (SPF) conditions in the same unit of the Forschungseinrichtungen für Experimentelle Medizin (FEM, Charité—University Medicine Berlin). In order to counteract physiological colonization resistance and thus assure stable gastrointestinal C. jejuni colonization [13 (link)], secondary abiotic mice with a depleted gut microbiota were generated as described earlier [13 (link), 54 (link)]. In brief, immediately post weaning 3-week-old mice were subjected to a 10-week course of broad-spectrum antibiotic treatment by adding ampicillin plus sulbactam (1 g/l; Ratiopharm, Germany), vancomycin (500 mg/l; Cell Pharm, Germany), ciprofloxacin (200 mg/l; Bayer Vital, Germany), imipenem (250 mg/l; MSD, Germany) and metronidazole (1 g/l; Fresenius, Germany) to the autoclaved drinking water (ad libitum). Two days before pathogenic challenge the antibiotic cocktail was replaced by autoclaved tap water to assure antibiotic washout.
4
Antibiotic Source Verification for Bacterial Testing
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The antibiotics for preparation of test plates were procured from the suppliers shown in parentheses: Amikacin, Ampicillin, Cefotaxime, Ceftriaxone, CiprOfloxacin, Gentamicin, Nitrofurantoin, Norfloxacin, Piperacillin/Tazobactam, Ofloxacin, (Sigma-Aldrich, USA), Amoxycillin/clavulanate (GlaxoSmithKline; Thane, India), Ceftazidime (VHB; Mumbai, India), Cefepime (Biocon; Bengaluru, India), Cefixime (GlaxoSmithKline; Thane, India), Colistin (Cipla; Pune, India), Erythromycin (Himedia; Mumbai, India), Imipenem (MSD; Mumbai, India), LevOfloxacin (Alkem; Chennai, India), Meropenem (Cipla; Pune, India), Tetracycline (GlaxoSmithKline; Thane, India), Tigecycline (Natco; Chennai, India), Vancomycin (Himedia; Mumbai, India).
5
Establishing Stable C. jejuni Colonization
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Female and male IL-10−/− mice (all in C57BL/6j background) were bred and maintained under specific pathogen free (SPF) conditions in the same unit of the Forschungseinrichtungen für Experimentelle Medizin (FEM, Charité - University Medicine Berlin). To order to assure stable gastrointestinal C. jejuni colonization and override physiological colonization resistance (Bereswill et al., 2011 (link)), secondary abiotic (i.e., gnotobiotic) mice virtually lacking an intestinal microbiota were generated (Heimesaat et al., 2006 (link); Bereswill et al., 2011 (link); Haag et al., 2012a (link)). In brief, immediately post weaning 3 weeks old mice were subjected to a 10-week course of broad-spectrum antibiotic treatment by adding ampicillin plus sulbactam (1 g/L; Ratiopharm, Germany), vancomycin (500 mg/L; Cell Pharm, Germany), ciprofloxacin (200 mg/L; Bayer Vital, Germany), imipenem (250 mg/L; MSD, Germany) and metronidazole (1 g/L; Fresenius, Germany) to the autoclaved drinking water (ad libitum). To assure antibiotic washout, the antibiotic cocktail was replaced by autoclaved tap water 2 days prior infection.
6
Gut Flora Depletion in IL-10 Knockout Mice
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IL-10−/− knockout mice (in C57BL/10 background, B10) were bred and maintained under specific pathogen-free (SPF) conditions in the facilities of the “Forschungsinstitut für Experimentelle Medizin” (FEM, Charité - Universitätsmedizin, Berlin, Germany). To eradicate the commensal gut flora, mice were transferred to sterile cages and treated by adding a mix of ampicillin (1 g/L; Ratiopharm), vancomycin (500 mg/L; Cell Pharm), ciprofloxacin (200 mg/L; Bayer Vital), imipenem (250 mg/L; MSD), and metronidazole (1 g/L; Fresenius) to the drinking water ad libitum starting at 3 weeks of age right after weaning (Heimesaat et al., 2006 (link); Haag et al., 2012a (link)). Age matched female mice were subjected to the quintuple antibiotic treatment for approximately 4 months before the infection experiments.
7
Antibiotic-Induced Gut Flora Depletion
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IL-10−/− mice (in C57BL/10 background, B10) were bred and maintained in the facilities of the “Forschungsinstitut für Experimentelle Medizin” (FEM, Charité - Universitätsmedizin, Berlin, Germany), under specific pathogen-free (SPF) conditions.
To eradicate the commensal gut flora, mice were transferred to sterile cages and treated by adding ampicillin (1 g/L; Ratiopharm), vancomycin (500 mg/L; Cell Pharm), ciprofloxacin (200 mg/L; Bayer Vital), imipenem (250 mg/L; MSD), and metronidazole (1 g/L; Fresenius) to the drinking water ad libitum as described earlier [20] (link) starting at 3 weeks of age right after weaning. Age matched female mice were subjected to the quintuple antibiotic treatment for 3–4 months before the infection experiment.
To eradicate the commensal gut flora, mice were transferred to sterile cages and treated by adding ampicillin (1 g/L; Ratiopharm), vancomycin (500 mg/L; Cell Pharm), ciprofloxacin (200 mg/L; Bayer Vital), imipenem (250 mg/L; MSD), and metronidazole (1 g/L; Fresenius) to the drinking water ad libitum as described earlier [20] (link) starting at 3 weeks of age right after weaning. Age matched female mice were subjected to the quintuple antibiotic treatment for 3–4 months before the infection experiment.
8
Antibiotic Modulation of Murine Microbiome
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C57BL/6j wildtype mice were reared and maintained under specific pathogen-free (SPF) conditions in the Forschungseinrichtungen für Experimentelle Medizin (FEM, Charité – University Medicine Berlin). At the age of 8 to 10 weeks, female mice were subjected to broad-spectrum antibiotic treatment. In brief, mice were transferred to sterile cages and treated with a quintuple antibiotic cocktail consisting of ampicillin plus sulbactam (1 g/l; Ratiopharm, Ulm, Germany), vancomycin (500 mg/l; Cell Pharm, Hannover, Germany), ciprofloxacin (200 mg/l; Bayer Vital, Leverkusen, Germany), imipenem (250 mg/l; MSD, Haar, Germany), and metronidazole (1 g/l; Fresenius, Bad Homburg, Germany) via the drinking water ad libitum for 8 weeks [14 (link), 15 (link)]. Cultural and culture-independent (i.e., 16S rRNA based molecular) quality control measures revealed virtual absence of bacteria in fecal samples as described earlier [15 (link), 16 (link)]. In one group of thus generated secondary abiotic (i.e., gnotobiotic) mice, the antibiotic cocktail was replaced by sterile water three days before P. aeruginosa infection, whereas in another group, antibiotic treatment was continued until the end of the experiment. Sex- and age-matched conventionally colonized mice served as antibiotics-untreated control group.
9
Antibiotic Susceptibility Testing via Agar Dilution
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The minimum inhibitory concentrations (MIC) of several antibiotics were determined by the agar dilution method using Muller-Hinton agar II (Difco, Franklin Lake, NJ, USA) according to the method recommended by the Japanese Society of Chemotherapy. The antibiotics used were chloramphenicol 8–1024 µg/mL (Wako, Japan), ciprofloxacin 0.0625–32 µg/mL (LKT Laboratories, St Paul, MN, USA), tetracycline hydrochloride 0.5–128 µg/mL (Wako, Japan), amikacin sulfate 0.25–128 µg/mL (Wako, Osaka, Japan), aztreonam 0.25–32 µg/mL (MP Biomedicals, USA), imipenem 0.125–32 µg/mL (MSD K.K., Tokyo, Japan), rifampicin 2–512 µg/mL (Tokyo Chemical Industry, Tokyo, Japan), and minocycline hydrochloride 1–256 µg/mL (Tokyo Chemical Industry, Tokyo, Japan). The standard laboratory strain, PAO1S, and representative clinical isolated strain, 8380 strain, were used as control. All experiments were performed in triplicate.
10
Generation of Abiotic Mouse Model
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Female C57BL/6j mice were bred and maintained under SPF conditions in the Forschungsinstitute für Experimentelle Medizin (Charité – University Medicine, Berlin, Germany). Secondary abiotic mice with a virtually depleted microbiota were generated as described previously (Heimesaat et al., 2006 (link)). In brief, 8 weeks old mice were transferred into sterile cages and subjected to a broad-spectrum antibiotic treatment for 8 weeks by adding ampicillin plus sulbactam (1 g/L; Ratiopharm, Germany), vancomycin (500 mg/L; Cell Pharm, Germany), ciprofloxacin (200 mg/L; Bayer Vital, Germany), imipenem (250 mg/L; MSD, Germany) and metronidazole (1 g/L; Fresenius, Germany) to the drinking water (ad libitum). Cultural and culture-independent (i.e., 16S rRNA based molecular) quality control measures revealed virtual absence of bacteria in fecal samples as described earlier (Ekmekciu et al., 2017 (link)).
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