All materials were used without further purification unless otherwise noted. Tannic acid, vancomycin HCl (hereafter referred to as vancomycin unless otherwise noted), and poly-l -lysine (PLL, 30–70 kDa) were obtained from Sigma-Aldrich (St. Louis, MO). Poly(β-l -malic acid) (PMLA, 40 kDa) was cultured and purified as previously described from Physarum polycephalum.13 Bovine thrombin (high purity) was obtained from Biopharm Laboratories (Bluffdale, UT). Cation-adjusted Mueller Hinton broth (CaMHB) was obtained from Becton Dickenson (Franklin Lakes, NJ). All other chemicals unless otherwise noted, were obtained from Sigma-Aldrich (St. Louis, MO).
Vancomycin hcl
Manufactured by Merck Group
Sourced in Australia
Vancomycin HCl is a sterile, crystalline powder used as a reference standard for the identification and quantitative determination of vancomycin in pharmaceutical and biological samples. It is a glycopeptide antibiotic that disrupts cell wall synthesis in gram-positive bacteria.
Automatically generated - may contain errors
Lab products found in correlation
Colistin sulfate, by Merck Group (3 mentions)
Cls3640, by Corning (3 mentions)
Metronidazole, by Merck Group (2 mentions)
Ampicillin sodium salt, by Merck Group (2 mentions)
Gentamycin sulfate, by Merck Group (2 mentions)
Neomycin sulfate, by Merck Group (2 mentions)
Poly l lysine (pll), by Merck Group (1 mentions)
Tannic acid, by Merck Group (1 mentions)
Cation adjusted mueller hinton broth, by BD (1 mentions)
Ammonium hydroxide nh4oh, by Thermo Fisher Scientific (1 mentions)
Phosphate buffered saline (pbs), by American Type Culture Collection (1 mentions)
Ciprofloxacin, by Merck Group (1 mentions)
Tetraethyl orthosilicate teos, by Merck Group (1 mentions)
Hyaluronic acid, by Lifecore Biomedical (1 mentions)
Imipenem, by Merck Group (1 mentions)
Acetic acid hoac, by Merck Group (1 mentions)
Gentamycin sulfate 600 iu mg, by Thermo Fisher Scientific (1 mentions)
Type 70 ti fixed angle rotor, by Beckman Coulter (1 mentions)
Nanosight analysis, by Malvern Panalytical (1 mentions)
8 protocols using vancomycin hcl
1
Tannic Acid-Vancomycin Antimicrobial Formulation
2
Antimicrobial Susceptibility Assay Protocol
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For the all the bacterial assays, each strain was cultured in Cation-adjusted Mueller Hinton broth (CAMHB; Bacto Laboratories 212322) at 37 °C overnight. A sample of each culture was then diluted 40-fold in fresh CAMHB and incubated at 37 °C for 1.5–3 h. The resultant mid-log phase cultures were diluted with CAMHB (CFU mL–1 measured by OD600), then added to each well of the compound-containing plates (384-well non-binding surface (NBS) plates; Corning CLS3640), giving a cell density of 5 × 105 CFU mL–1 and a total volume of 50 μL. Plates were covered and incubated at 37 °C for 18 h without shaking. Inhibition of bacterial growth was determined measuring absorbance at 600 nm (OD600), using media only as negative control and bacteria without inhibitors as positive control. MIC values were determined as the lowest concentration at which the growth was inhibited at ≥80%. Colistin sulfate (Sigma C4461) and vancomycin HCl (Sigma 861987) were used as internal controls on each plate for Gram-negative and Gram-positive bacteria, respectively.
3
Antimicrobial Susceptibility Assay for Bacterial Strains
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For the bacterial assays, each bacterial strain was cultured in Cation-Adjusted Mueller Hinton Broth (CAMHB; Bacto Laboratories 212322) at 37 °C overnight. Bacterial strains tested were methicillin resistant S. aureus (methicillin-resistant Staphylococcus aureus - MRSA) ATCC 43300, E. coli ATCC 25922, K. pneumoniae K6/ESBL ATCC 700603, P. aeruginosa ATCC 27853, A. baumannii ATCC 19606. A sample of each culture was then diluted 40-fold in fresh CAMHB and incubated at 37 °C for 1.5–3 h. The resultant mid-log phase cultures were diluted with CAMHB (CFU/mL measured by OD600), then added to each well of the compound-containing plates (384-well non-binding surface (NBS) plates; Corning CLS3640), giving a final cell density of 5 × 105 CFU/mL and a total volume of 50 μL. Plates were covered and incubated at 37 °C for 18 h without shaking. Inhibition of bacterial growth was determined by measuring absorbance at 600 nm (OD600), using media only as negative control and bacteria without inhibitors as positive control. MIC values were determined as the lowest concentration at which the growth was inhibited by ≥ 80% (equivalent to no visible growth by eye). Colistin sulfate (Sigma Aldrich, Castle Hill, Australia C4461) and vancomycin HCl (Sigma Aldrich, 861987) were used as positive inhibitor controls on each plate for Gram-negative and Gram-positive bacteria, respectively.
4
Silk Fibroin Hydrogel Synthesis
5
Comparative Vancomycin HCl MIC Analysis
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Example 9
Introduction: The purpose of the study was to compare the MICs obtained with the formulation of vancomycin HCL of the invention (Cutis) to formulations obtained using a vancomycin HCL reference standard powder obtained from Sigma. The Example also involves the analysis of a solution or diluent to determine if it has antimicrobial activity at the concentrations present in the agar dilution test.
6
Isolation of Intestinal Extracellular Vesicles
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The intestinal EVs were prepared from small intestine lumen contents of ageing WT mice with sterile tools. Debris and dead cells in the lumen contents were removed by centrifugation at 1,000 x g for 10 min and then passed through a 0.2 µm filter. Then, the supernatant was added with a mixture of antibiotics (0.5 mg/mL vancomycin HCl, 1 mg/mL ampicillin sodium salt, 1 mg/mL metronidazole, 1 mg/mL neomycin sulfate, and 1mg/mL gentamycin sulfate; Sigma) and then ultracentrifuged at 100,000 x g for 4 hours at 4°C with a Type 70 Ti fixed-angle rotor (Beckman Coulter). The EV-containing pellet was resuspended in 1 mL of sterile PBS and passed through a 0.2 µm filter to remove large particles in a sterile hood. The particle size and concentration of intestinal EVs were detected by NanoSight analysis (Malvern Instruments).
7
Bacterial Growth Inhibition Assay
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For the all the bacterial assays,
each bacterial strain was cultured in cation-adjusted Mueller Hinton
broth (CAMHB; Bacto Laboratories 212322) at 37 °C overnight.
A sample of each culture was then diluted 40-fold in fresh CAMHB and
incubated at 37 °C for 1.5–3 h. The resultant mid-log
phase cultures were diluted with CAMHB (CFU/mL measured by OD600) and then added to each well of the compound-containing
plates (384-well non-binding surface (NBS) plates; Corning CLS3640),
giving a cell density of 5 × 105 CFU/mL and a total
volume of 50 μL. Plates were covered and incubated at 37 °C
for 18 h without shaking. Inhibition of bacterial growth was determined
by measuring the absorbance at 600 nm (OD600) using media
only as negative control and bacteria without inhibitors as positive
control. MIC values were determined as the lowest concentration at
which the growth was inhibited by ≥80% (equivalent to no visible
growth by the eye). Colistin sulfate (Sigma C4461) and vancomycin
HCl (Sigma 861987) were used as internal controls on each plate for
Gram-negative and Gram-positive bacteria, respectively. All compounds
were tested as two technical replicates in two independent biological
assays, n = 4 final data.
each bacterial strain was cultured in cation-adjusted Mueller Hinton
broth (CAMHB; Bacto Laboratories 212322) at 37 °C overnight.
A sample of each culture was then diluted 40-fold in fresh CAMHB and
incubated at 37 °C for 1.5–3 h. The resultant mid-log
phase cultures were diluted with CAMHB (CFU/mL measured by OD600) and then added to each well of the compound-containing
plates (384-well non-binding surface (NBS) plates; Corning CLS3640),
giving a cell density of 5 × 105 CFU/mL and a total
volume of 50 μL. Plates were covered and incubated at 37 °C
for 18 h without shaking. Inhibition of bacterial growth was determined
by measuring the absorbance at 600 nm (OD600) using media
only as negative control and bacteria without inhibitors as positive
control. MIC values were determined as the lowest concentration at
which the growth was inhibited by ≥80% (equivalent to no visible
growth by the eye). Colistin sulfate (Sigma C4461) and vancomycin
HCl (Sigma 861987) were used as internal controls on each plate for
Gram-negative and Gram-positive bacteria, respectively. All compounds
were tested as two technical replicates in two independent biological
assays, n = 4 final data.
8
Antibiotic Impact on Ageing Mice
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Ageing WT mice (100-108wks old) were subject to oral gavage (twice per week; 200 µL per mouse) with a mixture of broad-spectrum antibiotics (0.5 mg/mL vancomycin HCl, 1 mg/mL ampicillin sodium salt, 1 mg/mL metronidazole, 1 mg/mL neomycin sulfate, and 1mg/mL gentamycin sulfate; Sigma) (48 (link)). Control mice were treated with water alone. The ageing WT mice in the antibiotic withdrawal group were fed ad libitum without antibiotic treatment for another 2 wks.
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