Transcutol

Garlic oil was donated by the Verywell Health Company (New York, NY, USA). Tween 20 and Span 20 were purchased from the BASF SE Chemicals Company (Ludwigshafen, Germany). Apple cider vinegar was procured from Jiangxi Origin Aromatics Co., Ltd. (Xi’an, China). Minoxidil was gifted by Qingdao Sigma Chemical Co., Ltd. (Qingdao, China). Transcutol was donated by Gattefosse (Saint-Priest, France). Methanol, acetonitrile, and phosphate HPLC-grade buffers were purchased from Sigma Aldrich (St. Louis, MO, USA).

Female Balb/cAnNCrl mice were purchased from Charles River Laboratories (Wilmington, MA, USA) or bred at the laboratory mouse facility of the faculty of medicine, University of Rijeka, Rijeka, Croatia. Mice were maintained under specific pathogen-free conditions and utilized between 5 and 9 weeks of age. Caging was performed in groups of six mice maximum and body weight was monitored daily. Animals were infected with MCMV-UL97 [24 (link)] at 1.0 × 10⁸ PFU intraperitoneally (i.p.) in a final volume of 100 µL PBS or remained mock-infected at d 0. Antiviral compounds were administered daily (d 0 to d 3), either via oral gavage using feeding needles (p.o.) or via the intraperitoneal route (i.p). A solution of 20% PHOSAL^® 50 PG (Lipoid GmbH, Ludwigshafen, Germany) in PBS was used as solvent and vehicle control for the p.o.-treated cohort; a solution of 2.5% DMSO + 5% transcutol (Gattefossé, Saint-Priest Cedex, France) in 0.9% NaCl in PBS was used as solvent and vehicle control for the i.p.-treated cohort. Mice were sacrificed at 4 d p.i., and spleen, heart, liver and lung were dissected, subdivided into experimental bits and stored at −80 °C. Experimental protocols were reviewed and approved by the Regierung von Unterfranken, Würzburg, Germany (permit 55.2-2532-2-416; 2017–2022).

Before leaving the sorting facility, animals were moved again through a chute. Five milliliters of SecureCattle^® (Signs—Irsea Group, Apt, France) (pheromone group) or 5 mL of transcutol (Diethylene glycol monoethyl ether—Gattefossé, Saint-Priest, France) (control group), a high-purity solvent and powerful solubilizer associated with skin penetration enhancement in topical dosage forms, were deposited on the coat in the middle of the forehead—at the crossing point of two imaginary lines drawn between the eyes and the center of the base of the opposite horns. An Injecmaster^® automatic syringe without any needle was used to perform the pour-on administration (Génia, St. Hilaire de Chaléons, France). From there, contacts between young bulls from pheromone and control groups were prevented using an empty pen between groups in some fattening units and double fences with empty spaces between fences in the other groups.

All the reagents and solvents used were analytical or chromatographic grade. The materials used were as follows: chitosan (deacetylation degree > 75%) (Sigma-Aldrich^®, MO, USA); Transcutol^® (ethoxydiglycol) (Gattefossé^®, Lyon, France); methylparaben, glacial acetic acid, ethanol, ethyl acetate, n-hexane, and methanol (Synth^®, São Paulo, Brazil); activated carbon (specification type in patent number BR102014022486-6); diatomaceous earth (Celite^® 545) (Nuclear^®, São Paulo, Brazil); eutectic mixture of 2.5% lidocaine and 2.5% prilocaine (5% EMLA^® cream—AstraZeneca^®, São Paulo, Brazil); chromatography grade methanol (J.T. Baker^®, PA, USA).

The BCT bulk powder ingredient was supplied by Henrikang Biotech Co., Ltd. (Xi’an, China). Transcutol, Labrafac, Isostearyl Isostearate, Labrasol, Lauroglycol 90, Labrafil M 1944 CS, Plurol oleique, and Capryol 90 were acquired from Gattefossé (Saint-Priest, France). Dimethyl sulfoxide (DMSO), Tween-80, and oleic acid were provided by Panreac Química SA (Barcelona, Spain); limonene, α-pinene, nonane, 1-decanol 99%, octanoic acid, lauryl sulfate, sebacic acid, castor oil, and phosphate-buffered solution pH 7.4 were purchased from Sigma Aldrich (St. Louis, MO, USA); Surfadone LP 100, N-ethyl pyrrolidone (NEP), and N-methyl pyrrolidone were obtained from ISP (West Yorkshire, UK). Perhydro Squalene was acquired from Fagron Iberica (Terrassa, Spain), liquid paraffin was supplied by Roig Farma, (Terrassa, Spain), and distilled water and purified water were obtained using a Station 9000 purification unit.

Febuxostat (FBX) powder was a kind gift from SPIMACO Addwaeih’s. (Riyadh, Saudi Arabia). Propylene glycol and clove oil were purchased from TEDIA Company, Inc. (Fairfield, OH, USA). Olive oil, linoleic acid 60% and isopropyl myristate 96% were purchased from Acros Organics (Morris Plains, NJ, USA). Microcrystalline cellulose (Avicel^® PH-101), fumed silica (0.007 µm), gelatin, Tween 80, Tween 20, polyethylene glycol (PEG) 200 and 400, triacetin, oleic acid, Lauroyl polyoxylglycerides (Gelucire^® 44/14, hydrogenated coconut oil with PEG 32), mannitol, castor oil, methanol, polyethylene glycol 40 stearate (Myrj^® 52), glycerin, hydroxypropyl methylcellulose (HPMC), xylitol, croscarmellose sodium (Ac-Di-Sol^®), and starch were purchased from Sigma-Aldrich (St. Louis, MO, USA). Propylene glycol dicarprylate, Labrafil^® M1944, Labrafac^® WL1349, Labrasol^®, Transcutol^®, and Kolliphor^® were purchased from Gattefosse (Saint-Priest, France). Lactose was obtained from the Spectrum chemical manufacturing corporation (Gardena, CA, USA). All other chemicals and solvents were of analytical grades.