Sodium taurocholate

Olive oil, bovine gall powder, triolein, lecithin (egg yolk), sodium taurocholate, catechin mixture, and oleic acid were purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). Lipase, glyceride monooleate, and cholesterol were purchased from Sigma-Aldrich Corporation (Tokyo, Japan). Epigallocatechin gallate hydrate (EGCG) was purchased from Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan). oleic acid-glyceryl monooleate mixture was purchased from MP Biomedicals (Tokyo, Japan). Indigestible dextrin was purchased from Matsutani Chemical Industry Co., Ltd. (Hyogo, Japan). Pectin was purchased from Sansho Co., Ltd. (Osaka, Japan). IMD was produced by Hayashibara Co., Ltd. (Okayama, Japan)[12 (link)].

A culture assay was performed to determine the efficacy of the combination of ABZ and ATV. The protoscoleces obtained were cultured in Connaught Medical Research Laboratories 1066 medium (Gibco, Grand Island, NY, USA) containing 0.5% (w/v) yeast extract (Difco Laboratories, Detroit, MI, USA), 23 mM 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid, 0.5% (w/v) D (+)-glucose, 0.4 mM sodium taurocholate (Wako Pure Chemical Industries, Osaka, Japan), 57 mM sodium hydrogen carbonate, 2 mM L-glutamine (Gibco), 100 U/mL penicillin, and 100 μg/mL streptomycin (Gibco). Half of the medium was changed on day 3. For anaerobic cultures, six-well plates were sealed in plastic containers with oxygen-detecting agents and oxygen scavengers (Aneromeito®, Nissui Pharmaceutical, Tokyo, Japan) to maintain the oxygen concentration under 0.3% at 37 °C. The protoscoleces were treated with ABZ (Tokyo Chemical Industry, Tokyo, Japan) and/or ATV (Tokyo Chemical Industry) at a final concentration of 50 μM in the culture medium, and the duration of parasite elimination was determined. The control group was supplemented with 0.5% (v/v) dimethyl sulfoxide (DMSO), and all conditions were assayed in triplicate. The viability of protoscoleces was determined by microscopic observation of more than 170 protoscoleces per well using the trypan blue exclusion test.

Vancomycin hydrochloride, polymyxin B sulfate, lithocholic acid, sodium deoxycholate, sodium taurocholate, and lysyl endopeptidase were purchased from Wako Pure Chemical Industries (Osaka, Japan). QIAamp Fast DNA Stool Mini Kit was from Qiagen (Hilden, Germany). Taq DNA polymerase containing 10 × Standard buffer (Taq) and dNTPs were obtained from BioAcademia (Osaka, Japan) and synthesised PCR primers were obtained from FASMAC (Kanagawa, Japan). Sodium taurolithocholate and sodium taurochenodeoxycholate were purchased from Sigma-Aldrich (St. Louis, MO, USA). Taurodeoxycholic acid sodium salt was purchased from Nacalai Tesque (Kyoto, Japan). Tauro β-muricholic acid sodium salt was purchased from Steraloids (Newport, RI, USA). Taurocholic acid-d5 (TCA-d5) sodium salt was purchased from Toronto Research Chemicals (Toronto, Ontario, Canada). Triglyceride Quantification Assay kit was purchased from Abcam (Cambridge, UK). Plasma Membrane Protein Extraction Kit was obtained from BioVision (Milpitas, CA, USA). Pierce BCA Protein Assay Kit was obtained from Thermo Fisher Scientific (Waltham, MA, USA). Sequencing-grade modified trypsin (frozen) was obtained from Promega (Madison, WI, USA). Synthesised isotope-labelled peptides were obtained from Sigma-Aldrich. Other reagents were commercially available products of reagent or analytical grade.