Ruthenium red

Yeast cells isolated from the brines and olives on RBCA were selected on the basis of colony morphology to test for PGA activity. The yeast isolates were first grown for 24 h in YM containing 1.5% (wt/vol) pectin (Spectrum Chemical Manufacturing Corp., Gardena, CA) and were then dispensed into preformed wells in agar plates containing polygalacturonic acid prepared as previously described (4 (link)). The plates were incubated at 30°C for 24 h and stained with 0.05% ruthenium red (Acros Organics, Geel, Belgium) for 30 min, and the PGA activity was quantified by measuring the radius of the zone of clearing around the wells (28 (link)). Purified PGA at a concentration of 0.001 mg/ml (Sigma-Aldrich, St. Louis, MO) was used as a positive control.

Histamine dihydrochloride and capsaicin (TRPV1 agonist) were purchased from Sigma-Aldrich (Steinheim, Germany). The 4-methylHistamine dihydrochloride (4-MH; H2R /H4R agonist), and histamine trifluoromethyl toluidine dimaleate (HTMT; H1R/H2R agonist) were obtained from Tocris Bioscience (Bristol, UK). ST-1006 (H4R agonist) and JNJ-7777120 (H4R antagonist) were synthesized and provided by Prof. Dr. H. Stark (Heinrich-Heine-Universität, Düsseldorf, Germany) [13 (link),14 (link)]. Allylisothiocyanate (AITC; TRPA1 agonist) and ruthenium red (TRP channel inhibitor) were purchased from Acros Organics, Morris Plaines, NJ, USA. Capsazepine (TRPV1 inhibitor), SB366791 (TRPV1 inhibitor) and HC-030031 (TRPA1 inhibitor) were obtained from Cayman Chemicals (Ann Arbor, MI, USA). Diphenhydramine hydrochloride (H1R antagonist) was purchased from West Ward Pharmaceuticals (Eatontown, NJ, USA).