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Research Article

Inhibition by Resistant Starch of Red Meat–Induced Promutagenic Adducts in Mouse Colon

Jean Winter, Laura Nyskohus, Graeme P. Young, Ying Hu, Michael A. Conlon, Anthony R. Bird, David L. Topping and Richard K. Le Leu
Jean Winter
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Laura Nyskohus
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Graeme P. Young
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Ying Hu
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Michael A. Conlon
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Anthony R. Bird
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David L. Topping
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Richard K. Le Leu
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DOI: 10.1158/1940-6207.CAPR-11-0176 Published November 2011
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    Figure 1.

    The average sum of O6MeG staining intensity for each dietary treatment (A). Resistant starch (RS) supplemented in the diet (B) significantly reduced DNA adduct formation (P < 0.01) as compared with those diets without resistant starch. Compared with casein, red meat (C) significantly increased adduct formation (P < 0.018) irrespective of protein amount. There was no effect of protein amount (D) on DNA O6MeG staining intensity irrespective of the protein source. Data are presented as means ± SE.

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    Figure 2.

    Average distribution of O6MeG formation (A) for each dietary treatment along the length of the crypt. The highest cell number represents the top of the crypt. Data represented as RoB ratio, a higher value represents a higher intensity of positive staining in the cell nucleus. Light microscopic image of distal colonic crypt (B) at 20× optical zoom showing distribution of O6MeG staining intensity. RS, resistant starch.

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  • Table 1.

    Composition of experimental diets (g/100 g in diet)

    Low caseinHigh caseinLow casein + RSLow meatHigh meatHigh meat + RS
    Caseina17.635.335.3———
    Red meata———20.4340.940.9
    Corn starch1010—1010—
    High amylose maize starch——10——10
    Sucrose45.4827.7827.7846.3929.6929.68
    Sunflower seed oil12.512.512.512.512.512.5
    Lardb7.57.57.53.76——
    α-Cellulose222222
    l-Cysteine0.30.30.30.30.30.3
    Choline0.120.120.120.120.120.12
    Mineralsc3.53.53.53.53.53.5
    Vitaminsc1.01.01.01.01.01.0
    • ↵aEnough casein and red meat were added to reach a total protein content of 15% for the low protein diets and 30% for the high protein diets.

    • ↵bLard was added to the casein and low red meat diets to balance each diet for saturated fat and to give a total fat content of 20%.

    • ↵cAIN-76 vitamin and mineral mixtures with modified calcium at 0.5 mg/g, phosphorus at 3.6 mg/g, folic acid at 0.23 μg/g, and vitamin D3 at 0.11 IU/g.

  • Table 2.

    Effect of dietary protein amount, protein type, and resistant starch supplementation on fecal fermentation measuresa

    Treatment groupsPb
    Low caseinHigh caseinHigh casein + RSLow meatHigh meatHigh meat + RSProtein amountProtein typeRS
    Fecal pH7.8 ± 0.17.5 ± 0.16.9 ± 0.67.2 ± 0.17.6 ± 0.16.7 ± 0.10.910.09<0.0001
    Acetate, μmol/g16.8 ± 2.820.0 ± 3.428.0 ± 3.628.1 ± 4.625.3 ± 4.741.4 ± 3.90.760.07<0.0001
    Propionate, μmol/g4.7 ± 0.64.9 ± 0.617.9 ± 2.26.4 ± 0.76.9 ± 0.926.4 ± 2.10.210.01<0.0001
    Butyrate, μmol/g2.9 ± 0.63.5 ± 0.412.2 ± 1.73.9 ± 0.66.2 ± 1.717.4 ± 2.10.150.08<0.0001
    Total SCFAs, μmol/g24.0 ± 3.828.4 ± 4.358.1 ± 7.038.4 ± 5.638.4 ± 6.285.2 ± 7.20.910.039<0.0001
    Ammonia, μmol/mL6.5 ± 1.58.3 ± 1.94.5 ± 0.94.1 ± 0.36.3 ± 1.63.7 ± 0.40.190.150.05
    Phenol, μg/g1.5 ± 0.31.4 ± 0.30.2 ± 0.11.3 ± 0.31.9 ± 0.30.9 ± 0.30.290.53<0.0001
    p-Cresol, μg/g3.8 ± 0.43.9 ± 0.82.2 ± 0.15.9 ± 0.97.4 ± 0.86.2 ± 0.60.48<0.00010.16

    Abbreviation: RS, resistant starch.

    • ↵aValues expressed as means ± SE (n = 12).

    • ↵bUnivariate ANOVA was used to determine the main effect of protein amount, protein type, and resistant starch supplementation.

  • Table 3.

    Effect of dietary protein amount, protein type, and resistant starch supplementation on distal colon epithelial cell kineticsa

    Treatment groupsPb
    Low caseinHigh caseinHigh casein + RSLow meatHigh meatHigh meat + RSProtein amountProtein typeRS
    Crypt height (cells)22.3 ± 0.521.9 ± 0.623.1 ± 0.422.5 ± 0.622.3 ± 0.423.2 ± 0.40.620.590.03
    Proliferation (cells per crypt)3.7 ± 0.23.3 ± 0.23.7 ± 0.33.5 ± 0.23.5 ± 0.14.2 ± 0.40.390.850.04
    Apoptosis (cells per crypt)0.3 ± 0.10.2 ± 0.10.2 ± 0.030.3 ± 0.10.3 ± 0.10.1 ± 0.020.370.730.001

    Abbreviation: RS, resistant starch.

    • ↵aValues expressed as means ± SE (n = 12).

    • ↵bUnivariate ANOVA was used to determine the main effect of protein amount, protein type, and resistant starch supplementation.

  • Table 4.

    Relationships between fermentation products, epithelial kinetics, and adduct formationa

    Fecal pHAmmoniap-CresolsPhenolsAcetatePropionateButyrateTotal SCFAs
    Crypt height
     Pearson correlation−0.206−0.130−0.079−0.1000.2400.2150.1140.244b
     P0.090.300.520.410.050.090.400.05
    Proliferation
     Pearson correlation−0.024−0.159−0.026−0.1940.0590.1970.1770.163
     P0.850.210.830.110.640.120.180.20
    Apoptosis
     Pearson correlation0.224−0.175−0.0150.161−0.199−0.362c−0.325b−0.320b
     P0.070.160.900.190.110.0030.0130.01
    Sum of O6MeG
     Pearson correlation0.377c−0.0640.234b0.0460.029−0.214−0.234−0.096
     P0.0020.610.0490.700.810.090.070.45
    • ↵aCorrelation for fecal measures versus DNA adducts and epithelial cell kinetics was undertaken by Pearson bivariate, one-tailed, parametric correlation tests.

    • ↵bSignificant at P < 0.05.

    • ↵cSignificant at P < 0.01.

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Cancer Prevention Research: 4 (11)
November 2011
Volume 4, Issue 11
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Inhibition by Resistant Starch of Red Meat–Induced Promutagenic Adducts in Mouse Colon
Jean Winter, Laura Nyskohus, Graeme P. Young, Ying Hu, Michael A. Conlon, Anthony R. Bird, David L. Topping and Richard K. Le Leu
Cancer Prev Res November 1 2011 (4) (11) 1920-1928; DOI: 10.1158/1940-6207.CAPR-11-0176

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Inhibition by Resistant Starch of Red Meat–Induced Promutagenic Adducts in Mouse Colon
Jean Winter, Laura Nyskohus, Graeme P. Young, Ying Hu, Michael A. Conlon, Anthony R. Bird, David L. Topping and Richard K. Le Leu
Cancer Prev Res November 1 2011 (4) (11) 1920-1928; DOI: 10.1158/1940-6207.CAPR-11-0176
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