Intestinal microbiota were depleted using a cocktail of metronidazole (Acros Organics), neomycin trisulfate (Sigma), and ampicillin sodium salt (Sigma) (all 1 g/l) provided in drinking water for 1 week. Indicated groups received 200 μl fecal suspensions of SFB-monocolonized mice by oral gavage 1 week after ceasing antibiotics. For single antibiotic treatments, ampicillin sodium salt (1 g/l), metronidazole (1 g/l), vancomycin hydrochloride (0.5 g/l, Fisher Scientific), or tobramycin sulfate (1 g/l, Centrafarm) was added to drinking water for 8 weeks and refreshed once a week. Sucrose (6 g/l) was added to drinking water of all groups including controls during treatments.
Vancomycin hydrochloride
Manufactured by Thermo Fisher Scientific
Sourced in China, United States, United Kingdom
Vancomycin hydrochloride is a white to off-white crystalline powder. It is a glycopeptide antibiotic used for the treatment of various bacterial infections.
Automatically generated - may contain errors
Lab products found in correlation
Metronidazole, by Thermo Fisher Scientific (2 mentions)
Sfb absent mice, by Jackson ImmunoResearch (2 mentions)
Sfb present mice, by Taconic Biosciences (2 mentions)
Gentamicin sulfate, by Merck Group (2 mentions)
Ampicillin sodium salt, by Merck Group (1 mentions)
Ampicillin sodium salt, by Thermo Fisher Scientific (1 mentions)
Neomycin trisulfate, by Merck Group (1 mentions)
Metronidazole, by Sanofi (1 mentions)
Neomycin sulfate, by Merck Group (1 mentions)
Boron trifluoride etherate, by Merck Group (1 mentions)
Hplc grade methanol, by Thermo Fisher Scientific (1 mentions)
Acetonitrile, by Thermo Fisher Scientific (1 mentions)
Whatman, by Cytiva (1 mentions)
Ampicillin, by MP Biomedicals (1 mentions)
Difco tryptic soy agar, by BD (1 mentions)
Bbl trypticase soy broth, by BD (1 mentions)
Glacial acetic acid, by Merck Group (1 mentions)
Sodium acetate, by Merck Group (1 mentions)
Glycidyl methacrylate, by Merck Group (1 mentions)
Cortecs c18, by Waters Corporation (1 mentions)
8 protocols using vancomycin hydrochloride
1
Modulating Gut Microbiome with Antibiotics
2
Modulating Gut Microbiome Composition
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For co-housing experiments, age-matched SFB-absent mice (from Jackson
Laboratory) were co-housed with SFB-present mice (from Taconic Biosciences) in
sterilized cages for two weeks at a ratio of 2:3, with unrestricted access to
food and water. For SFB-gavaging experiments, four fecal pellets of SFB
mono-colonized mice (provided by Dan Littman) were dissolved in 20 ml sterile
PBS and filtered though a 100 μm cell strainer. 200 μl of fecal
suspensions were gavaged via oral route to 4 week-old female Jackson mice.
Control mice were gavaged with PBS. The SFB colonization was tested on day 7
following co-housing or SFB-gavaging. For ablation of intestinal bacteria,
Taconic-derived female mice were orally gavaged with vancomycin hydrochloride
(Fisher) (2.5 mg/kg) every two days, starting 7 days prior to breeding. Mouse
fecal pellets were collected and stored at −80 °C before and
after vancomycin treatments.
Laboratory) were co-housed with SFB-present mice (from Taconic Biosciences) in
sterilized cages for two weeks at a ratio of 2:3, with unrestricted access to
food and water. For SFB-gavaging experiments, four fecal pellets of SFB
mono-colonized mice (provided by Dan Littman) were dissolved in 20 ml sterile
PBS and filtered though a 100 μm cell strainer. 200 μl of fecal
suspensions were gavaged via oral route to 4 week-old female Jackson mice.
Control mice were gavaged with PBS. The SFB colonization was tested on day 7
following co-housing or SFB-gavaging. For ablation of intestinal bacteria,
Taconic-derived female mice were orally gavaged with vancomycin hydrochloride
(Fisher) (2.5 mg/kg) every two days, starting 7 days prior to breeding. Mouse
fecal pellets were collected and stored at −80 °C before and
after vancomycin treatments.
3
Modulating Gut Microbiome Composition
4
Antibiotic Cocktail for Commensal Microbiota Ablation
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For ablation of commensal microbiota, an antibiotic cocktail of 1 g/l each of Ampicillin (sodium salt, Teva), Neomycin sulfate (Sigma), Metronidazole (Sanofi), and 0.5 g/l Vancomycin hydrochloride (Fisher) was used49 (link). Antibiotics were added to the drinking water on a weekly basis for 4 weeks.
5
Vancomycin Hydrochloride Analytical Protocol
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Vancomycin hydrochloride (Fisher Scientific ® , China) was stored at -20°C in a refrigerator until use. Boron trif luoride etherate (Merck ® , Darmstadt), Potassium dihydrogen phosphate monohydrate (Fisher Scientific ® , India), Sodium hydroxide (Gainland ® Chemical Company, UK), Phosphoric acid (Raw Mat., UK), Water HPLC Grade (Labchem ® , USA), acetonitrile and methanol HPLC Grades (Fisher Scientific ® , UK) were used without further purification.
Chloroform (Across Organic ® , USA) and Methanol (AZ ® chem for chemicals) were used in the purification of methylated vancomycin.
Trichloroacetic acid (TCA) (Riedel-deHaёn ® ), perchloric acid (Riedel-deHaёn ® ), and dried blood spot collection cards (Whatman, Kent, UK) were used. Blank blood and samples from vancomycintreated patients were donated from Dr. Hadidi's lab.
Chloroform (Across Organic ® , USA) and Methanol (AZ ® chem for chemicals) were used in the purification of methylated vancomycin.
Trichloroacetic acid (TCA) (Riedel-deHaёn ® ), perchloric acid (Riedel-deHaёn ® ), and dried blood spot collection cards (Whatman, Kent, UK) were used. Blank blood and samples from vancomycintreated patients were donated from Dr. Hadidi's lab.
6
Microbiota Depletion in SPF Mice
7
Antimicrobial K-wire Coatings Development
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Stainless steel K-wires (1.6 mm in diameter, SKU 40-1569) were obtained from Sklar (West Chester, PA, USA). Vancomycin hydrochloride was purchased from Alfa Aesar (Tewksbury, MA, USA). Gentamicin sulfate, glycidyl methacrylate (GMA), oligo-ethylene-glycol methacrylate (OEGMA), lauryl methacrylate, methyl ethyl ketone (MEK), glacial acetic acid, AIBN initiator, inhibitor removers, sodium docusate, polylactic acid (P1691), calcium chloride, potassium chloride, sodium acetate, dichloromethane (DCM), o-phtalaldehyde (OPA), and phosphate buffer saline (PBS) were purchased from Millipore Sigma (St. Louis, MO, USA). A synthetic human femur bone model (#3414) was purchased from Sawbones (Vashon Island, WA, USA). Chitosan and chitosan lactate were provided as a gift by Tidal Vision USA (Bellingham, WA, USA). Difco™ Tryptic Soy Agar and BBL™ Trypticase™ Soy Broth (TSB) were purchased from BD (Franklin Lakes, NJ, USA). Staphylococcus aureus (ATCC 33591, KWIK-STIKTM Plus) was obtained from Microbiologics (Saint Cloud, MN, USA).
8
HPLC Analysis of Vancomycin Hydrochloride
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Methanol, acetonitrile and triethylamine (TEA) were HPLC grade from Sigma-Aldrich (Dorset, UK). Vancomycin hydrochloride (VCN) was purchased from Alfa Aesar (Lancashire, UK).
Vancocin ® 500 mg powder (Vancomycin hydrochloride) solution for infusion and oral solution/gavage was obtained from Flynn Pharma Ltd. (Hertfordshire, UK). Sodium dihydrogen phosphate dihydrate was obtained from VWR International (Leuven, Belgium).
Orthophosphoric acid was purchased from Amresco (Ohio, USA). Ultrapure water was obtained from a water purification system Elga PURELAB DV 25 (Veolia Water Systems, Ireland). HPLC column Cortecs ® C18 (4.6 × 150 mm, 2.7 μm particle size) were purchased from Waters (Dublin, Ireland). All other chemical reagents were from Sigma-Aldrich (Dorset, UK). All reagents were of analytical grade.
Vancocin ® 500 mg powder (Vancomycin hydrochloride) solution for infusion and oral solution/gavage was obtained from Flynn Pharma Ltd. (Hertfordshire, UK). Sodium dihydrogen phosphate dihydrate was obtained from VWR International (Leuven, Belgium).
Orthophosphoric acid was purchased from Amresco (Ohio, USA). Ultrapure water was obtained from a water purification system Elga PURELAB DV 25 (Veolia Water Systems, Ireland). HPLC column Cortecs ® C18 (4.6 × 150 mm, 2.7 μm particle size) were purchased from Waters (Dublin, Ireland). All other chemical reagents were from Sigma-Aldrich (Dorset, UK). All reagents were of analytical grade.
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