Carboxymethylcellulose cmc

In ordinary saline, indomethacin (Sigma-Aldrich, St. Louis, MO, USA) was dissolved. The Arab Company for Pharmaceuticals and Medicinal Plants, Egypt, provided the standardized G. biloba leaf extract (EGb 761) as a pure powder (Batch No.: 510421, Registration No.: 888/2011). It was dissolved to a final concentration of 40 mg/mL in 0.25% carbox-ymethylcellulose (CMC; Sigma-Aldrich, St. Louis, MO, USA) in saline. EGb 761 was heated and vortexed to assure its solubility before being stored at 4 C for 24 h prior to administration. According to Chassagne et al. [81 (link)], voucher specimens (GEO20494, GEO20496, and GEO20497) are kept at the Emory University Herbarium in Atlanta, Georgia, in the United States [87 (link)].

The human epithelial cell lines CaCo2 and INT407 were used for C. jejuni infections. Medium used for growth of the cell lines was as directed by the ATCC. Cells were seeded into 24-well tissue culture plates at semiconfluence at ∼5 × 10⁵ and ∼1 × 10⁵ cells/ml for CaCo2 and INT407 cells, respectively. Infections were carried out as described (11 (link)). To test the levels of invasion in media of higher viscosity to mimic intestinal mucus, carboxymethylcellulose (CMC) (Sigma) was used. Infections were carried out as above using the INT407 cell line. The bacterial inoculum was added in MEM containing 0, 0.6 (141 centipoise (cP)), 1, and 2% CMC. Levels of adhered and invaded bacteria at the 1 and 3 h time points were determined by washing and lysing the cells and plating for cfu/ml as described (11 (link)).

The reagents used in this study for CoFe₂O₄ synthesis are: CoCl₂•6H₂O (ACS reagent, 99% Sigma-Aldrich, St. Louis, MO, USA), FeCl₂•6H₂O (Fisher Scientific, laboratory reagent grade, Waltham, MA, USA), NaOH (ROTH 98%, Carl Roth, Karlsruhe, Germany), carboxymethyl cellulose (CMC) (Sigma-Aldrich, St. Louis, MO, USA). The materials used in the synthesis of the CoFe₂O₄@HaP nanocomposite are: Ethanol (99.8% ROTH, Carl Roth, Karlsruhe, Germany), Hexadecyltrimethylammonium bromide (CTAB, Sigma Aldrich, St. Louis, MO, USA), H₃PO₄ (85% ROTH, Carl Roth, Karlsruhe, Germany), NH₄OH (25% CHIM REACTIV SRL, Bucharest, Romania), Ca(NO₃)₂ (Sigma-Aldrich, St. Louis, MO, USA).

LC-MS grade water (catalog no. W6–212) and chloroform (catalog no. C6704–4) were purchased from Fisher Scientific (Hampton, NH). Carboxymethylcellulose (CMC; catalog no. C4888) was purchased from Sigma-Aldrich (St. Louis, MO). Lipid standard D-lactosyl-ß-1,1′ N-palmitoyl-D-erythro-sphingosine (LacCer(d34:1), catalog no. 860576 P) was purchased from Avanti Polar Lipids, Inc. (Alabaster, AL). SEM reagents glutaraldehyde (catalog no. 16020), paraformaldehyde (catalog no. 15710), sodium cacodylate buffer (catalog no. 11652), anhydrous ethanol (catalog no. 15055), and osmium tetroxide (catalog no. 19170) were all purchased from Electron Microscopy Sciences (Hatfield, PA). Conductive silver paint (catalog no.16062) was purchased from Ted Pella, Inc. (Redding, CA).

RPG (purity > 98%), QCT (purity ≥ 95%), and ketoconazole (used as an internal standard; purity ≥ 98%) were purchased from Tokyo Chemical Industry Co. (Tokyo, Japan) Ethanol, dimethyl sulfoxide, polyethylene glycol 400 (PEG 400), carboxymethyl cellulose (CMC), potassium phosphate monobasic/dibasic, β-Nicotinamide adenine dinucleotide phosphate (NADPH), and phosphate-buffered saline (PBS) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Pooled male Sprague-Dawley rat plasma and pooled male human plasma were purchased from Innovative Research, Inc. (Novi, MI, USA) Pooled HLM and RLM were purchased from BD-Genetech (Woburn, MA, USA).

The evaluation of plaque-forming reduction (PFR) was performed with supernatants obtained from infected–treated cervical and Vero cells. The supernatants were diluted (1:1000) to allow plaque quantification, and 100 µL/well was added to Vero cell cultures seeded in 24-well plates. After 1 h of incubation under gentle agitation every 15 min., the cultures were washed to remove noninternalized viruses and added to a semisolid medium containing 3% carboxymethyl cellulose (CMC; Sigma-Aldrich, St. Louis, MO, USA) for 7 days at 37 °C under a 5% CO₂ atmosphere. Next, the cultures were fixed with 10% formaldehyde (Thermo Fisher Scientific, Waltham, MA, USA) and stained with 0.04% violet crystal (Sigma-Aldrich, St. Louis, MO, USA). For calculation purposes, we used (i) the percentage of plaque-forming reduction by the formula [1- (number of lysis plates in the treated/number of lysis plates in 0.25%DMSO)] × 100 [44 (link)]; (ii) the half-maximal effective concentration (EC₅₀), which reduces 50% of plaque-forming, by nonlinear regression analysis (dose–response curve) using GraphPad Prism 8.0 software (GraphPad Software Inc., San Diego, CA, USA); (iii) a selectivity index (SI), a ratio that measures the window between cytotoxicity (CC₅₀) and the antiviral effect (EC₅₀).