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3 protocols using tween 80

1

Bacterial Strains and Cell Line Protocol

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Neisseria gonorrhoeae clinical isolates (Table S1) used in this study were obtained from the CDC. S. aureus, MRSA, E. faecalis, E. faecium, and L. monocytogenes strains were obtained from the American Type Culture Collection (ATCC). E. coli BW25113 and JW25113 were obtained from the Coli Genetic Stock Center (CGSC), Yale University, USA. Media and reagents were purchased from commercial vendors: Brucella broth, chocolate II agar, cation-adjusted Mueller Hinton broth, tryptic soy broth (TSB), and tryptic soy agar (TSA) (Becton, Dickinson and Company, Cockeysville, MD, USA); yeast extract and dextrose (Fisher Bioreagents, Fairlawn, NJ, USA), proteose-peptone, nicotinamide adenine dinucleotide (NAD), agarose and tetracycline (Sigma-Aldrich, St. Louis, MO, USA); hematin, Tween 80, pyridoxal, linezolid and gentamicin sulfate (Chem-Impex International, Wood Dale, IL, USA); Dulbecco’s Modified Eagle Medium (DMEM), fetal bovine serum (FBS) and phosphate-buffered saline (PBS) (Corning, Manassas, VA, USA); and azithromycin (TCI America, Portland, OR, USA). Human colorectal adenocarcinoma epithelial cells (Caco-2) (ATCC HTB-37) was obtained from the American Type Culture Collection (ATCC) (Manassas, VA, USA). Compounds were synthesized from commercial sources in our laboratory.
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2

Investigating Cannabinoid Receptor Type 1 Antagonist Effects

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The peripherally restricted Cannabinoid Receptor Type 1 (CB1) antagonist, AM6545 (Sigma, St. Louis, MO, USA), was administered by IP injection at 10mg/kg in 2mL/kg. Vehicle consisted of 7.5% dimethyl sulfoxide (DMSO, Sigma, St. Louis, MO, USA), 7.5% tween 80 (Chem Impex Intl Inc., Wood Dale, IL, USA), 85% sterile saline (warmed in a bath sonicator for 30 minutes) 30 minutes prior (16:30 h) to the onset of behavioral analysis in feeding chambers. All control conditions were identical, except without drug present in the vehicle. The pharmacokinetics and half-life of AM6545 are not well-established, thus we chose to evaluate intakes over a period of 6 h from time of administration after the onset of the dark phase. Mice maintained on SD or WD were split into two groups and analyzed across two sessions of behavioral testing such that each subgroup received either vehicle or drug with 72h between administration.
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3

Antibiotic Susceptibility of Neisseria gonorrhoeae

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The N. gonorrhoeae reference strains were obtained from the WHO reference strain panel for global quality assurance and quality control of gonococcal AMR testing21 (link),41 (link). The rest of N. gonorrhoeae strains used were clinical isolates obtained from the CDC (Supplementary Table 1). The commensal vaginal bacterial strains were obtained from the Biodefense and Emerging Infections Research Resources Repository (BEI Resources) (Supplementary Table 2). All chemical compounds used in this study were obtained commercially from chemical vendors: auranofin (Chem-Impex International, Wood Dale, IL), sodium aurothiomalate, tetracycline and ciprofloxacin (Sigma-Aldrich, St. Louis, MO), aurothioglucose (USP reference standard Rockville, MD), ceftriaxone and cefixime (Acros Organics, NJ), and azithromycin (TCI America, Portland, OR).
Difco lactobacilli MRS broth, Brucella base broth and chocolate II agar (GC II Agar with hemoglobin and IsoVitaleX) were purchased from Becton, Dickinson and Company (Cockeysville, MD). Yeast extract and dextrose (Fisher Bioreagents, Fairlawn, NJ), proteose-peptone, nicotinamide adenine dinucleotide (NAD) and agarose (Sigma-Aldrich, St. Louis, MO), hematin solution, Tween 80, and pyridoxal (Chem-Impex International, Wood Dale, IL) were purchased from chemical vendors.
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