Compositions of diets (%).

<p>Compositions of diets (%).</p

ORAC values of soluble tannin powder and soluble tannin hydrolysate.

<p>Soluble tannin powder was hydrolyzed by heating at 90°C for 3 hours with 5 ml of a 1.2 N HCl–50% methanol solution, followed by dilution to 10 ml using 1.2 N HCl–50% methanol solution. The values are presented as means ± SEM (n = 3). *** <i>P</i> < 0.001 compared with non-hydrolyzed soluble tannin powder.</p

Gene expression of proinflammatory cytokines (IL-1ß, IL-6, TNF-α) and iNOS in BMDMs treated with soluble tannin hydrolysate.

<p>mRNA levels of IL-1ß (a), IL-6 (b), TNF-α (c), and iNOS (d) were determined by real-time PCR. BMDMs were pre-treated with soluble tannin hydrolysate (0, 30, and 100 μg/ml) for 1 hour followed by stimulation with three different strains of MAC (MAC-1, MAC-2, and MAC-3) for 6 hours. mRNA was extracted from BMDMs, reverse-transcribed into cDNA and qPCR performed. The values are presented as means ± SEM (n = 3–4). * <i>P</i> < 0.05, ** <i>P</i> < 0.01, *** <i>P</i> < 0.001 compared with no treatment with soluble tannin hydrolysate (0 μg/ml).</p

Proinflammatory cytokine (IL-1ß, IL-6, TNF-α) secretion by BMDMs treated with soluble tannin hydrolysate.

<p>BMDMs were pre-treated with soluble tannin hydrolysate (0, 30, and 100 μg/ml) for 1 hour followed by stimulation with three different strains of MAC (MAC-1, MAC-2, and MAC-3) for 6 hours. The supernatant was harvested and IL-1ß (a), IL-6 (b), and TNF-α (c) protein concentration was determined by ELISA. The values are presented as means ± SEM (n = 4). * <i>P</i> < 0.05, ** <i>P</i> < 0.01, *** <i>P</i> < 0.001 compared with no soluble tannin hydrolysate (0 μg/ml).</p

Analysis of soluble tannin and soluble tannin hydrolysate bacteriostatic activity against MAC.

<p>MAC strains (2×10² CFU each) were inoculated in culture medium with soluble tannin (hydrolysate). Soluble tannin (a) and soluble tannin hydrolysate (b) was added at 0, 30, and 100 μg/ml soluble tannin powder to each of three tubes and the CFU determined. The values are presented as means ± SEM (n = 4). * <i>P</i> < 0.05 when compared with no treatment with soluble tannin hydrolysate (0 μg/ml).</p

The effect of soluble tannin in a MAC-infected pulmonary granuloma model.

<p>Diets were started 1 week before MAC infection. All mice were analyzed at 8 weeks after MAC infection. (a) Microscopic morphology of granulomas in the H&E-stained lungs of MAC-infected BALB/c mice fed control and 2% soluble tannin diets. Dotted lines indicate the border of granulomas. Original magnification, ×40 and ×200. (b) Analysis of the size of lung granulomas between control and 2% soluble tannin diet. (c) MAC CFU counts in one lobe of lungs using Middlebrook 7H10 agar plates as described in the Materials and Methods. The values are presented as means ± SEM (uninfected mice, n = 5; MAC-infected mice, n = 8). * <i>P</i> < 0.05, *** <i>P</i> < 0.001 compared with MAC-infected mice fed the control diet.</p