Leslie D. Bourquin

Cyclooxygenase inhibitory and antioxidant cyanidin glycosides in cherries and berries

Anthocyanins from tart cherries, Prunus cerasus L. (Rosaceae) cv. Balaton and Montmorency; sweet cherries, Prunus avium L. (Rosaceae); bilberries, Vaccinum myrtillus L. (Ericaceae); blackberries, Rubus sp. (Rosaceae); blueberries var. Jersey, Vaccinium corymbosum L. (Ericaceae); cranberries var. Early Black, Vaccinium macrocarpon Ait. (Ericaceae); elderberries, Sambucus canadensis (Caprifoliaceae); raspberries, Rubus idaeus (Rosaceae); and strawberries var. Honeoye, Fragaria x ananassa Duch. (Rosaceae), were investigated for cyclooxygenase inhibitory and antioxidant activities. The presence and levels of cyanidin-3-glucosylrutinoside 1 and cyanidin-3-rutinoside 2 were determined in the fruits using HPLC. The antioxidant activity of anthocyanins from cherries was comparable to the commercial antioxidants, tert-butylhydroquinone, butylated hydroxytoluene and butylated hydroxyanisole, and superior to vitamin E, at a test concentration of 125 microg/ml. Anthocyanins from raspberries and sweet cherries demonstrated 45% and 47% cyclooxygenase-I and cyclooxygenase-II inhibitory activities, respectively, when assayed at 125 microg/ml. The cyclooxygenase inhibitory activities of anthocyanins from these fruits were comparable to those of ibuprofen and naproxen at 10 microM concentrations. Anthocyanins 1 and 2 are present in both cherries and raspberry. The yields of pure anthocyanins 1 and 2 in 100 g Balaton and Montmorency tart cherries, sweet cherries and raspberries were 21, 16.5; 11, 5; 4.95, 21; and 4.65, 13.5 mg, respectively. Fresh blackberries and strawberries contained only anthocyanin 2 in yields of 24 and 22.5 mg/100 g, respectively. Anthocyanins 1 and 2 were not found in bilberries, blueberries, cranberries or elderberries.

Tart cherry anthocyanins inhibit tumor development in ApcMin mice and reduce proliferation of human colon cancer cells

Anthocyanins, which are bioactive phytochemicals, are widely distributed in plants and especially enriched in tart cherries. Based on previous observations that tart cherry anthocyanins and their respective aglycone, cyanidin, can inhibit cyclooxygenase enzymes, we conducted experiments to test the potential of anthocyanins to inhibit intestinal tumor development in Apc(Min) mice and growth of human colon cancer cell lines. Mice consuming the cherry diet, anthocyanins, or cyanidin had significantly fewer and smaller cecal adenomas than mice consuming the control diet or sulindac. Colonic tumor numbers and volume were not significantly influenced by treatment. Anthocyanins and cyanidin also reduced cell growth of human colon cancer cell lines HT 29 and HCT 116. The IC(50) of anthocyanins and cyanidin was 780 and 63 microM for HT 29 cells, respectively and 285 and 85 microM for HCT 116 cells, respectively. These results suggest that tart cherry anthocyanins and cyanidin may reduce the risk of colon cancer.

Prevention of precancerous colonic lesions in rats by soy flakes, soy flour, genistein, and calcium

The main purpose of this research was to determine whether diets containing soy products would inhibit the early stages of azoxymethane-induced colon cancer in F344 rats. Additional objectives were to determine whether feeding starch instead of sucrose, feeding additional calcium (0.5% compared with 0.1%), or feeding a low-fiber powdered enteral formula would influence early colon carcinogenesis. Colon cancer was initiated with 2 injections of azoxymethane (15 mg/kg body wt) and a 12-wk dietary treatment period was started 1 wk after the second injection. Precancerous colon lesions were assessed as foci with aberrant crypts (FAC). The mean numbers of FAC were 133 [soy concentrate (low concentration of phytochemicals)], 111 (starch substituted for sucrose), 98 [full-fat soy flakes (whole soybeans)], 87 (defatted soy flour), 77 (0.015% genistein), and 70 (0.5% Ca). The soy flour and full-fat soy flake diets contained 0.049% genistein derivatives (primarily glycosides), but were less effective in inhibiting the formation of FAC than the diet containing 0.015% genistein (as the aglycone). Eating soybeans and soy flour may reduce the early stages of colon cancer.

Fermentation of various dietary fiber sources by human fecal bacteria

Abstract Eleven fiber-rich substrates were subjected to in vitro incubation with fecal bacteria from each of three adult male volunteers to assess substrate organic matter disappearance (OMD), short-chain fatty acid (SCFA) production, and the potential water holding capacity (PWHC) of fermented residues. Substrate OMD ranged from 83.3% (citrus pectin) to 5.8% (oat fiber). Total SCFA production (mmol/g substrate OM) was highly correlated to OMD and ranged from 9.64 (gum arabic) to 0.48 (oat fiber). Molar ratios of acetate:propionate produced during fermentation varied considerably among substrates, ranging from 1.5 for guar gum to 12.6 for citrus pectin. PWHC, an estimate of potential fecal bulking effects, was negatively related to OMD (r = −0.88) and ranged from 4.47 (oat fiber) to 0.71 (citrus pectin) g H 2 O/g substrate dry matter. In vitro fermentation techniques can be potentially useful predictors of in vivo responses to dietary fiber sources.

Fermentation of Dietary Fibre by Human Colonic Bacteria: Disappearance of, Short-Chain Fatty Acid Production from, and Potential Water-Holding Capacity of, Various Substrates

Several dietary fibre-rich substrates were fermented in vitro with human colonic bacteria obtained from each of three adult male subjects to assess the extent of substrate fermentation short-chain fatty acid (SCFA) production, and the potential effect of fermented residues on faecal bulk. Substrates tested were two varieties of oat hull fibre, gum arabic, carboxymethylcellulose (CMC), soy fibre, psyllium, and six blends containing oat fibre, gum arabic, and CMC in various proportions. All substrates contained greater than 900 g/kg of total dietary fibre except for CMC (816 g) and soy fibre (778 g). In vitro organic matter disappearance during fermentation was greatest for gum arabic (69.5%), intermediate for soy fibre (56.4%), and less than 20% for the two oat fibres, CMC, and psyllium. Averaged across substrates, acetate, propionate, and butyrate were produced in the molar proportion of 64:24:12. Potential water-holding capacity (PWHC) of substrates, a measure of faecal bulking potential, was greatest for CMC (13.5 g H2O/g substrate) and lowest for gum arabic (1.92 g) and soy fibre (1.71 g). Organic matter disappearance and SCFA production of blends were directly proportional to their gum arabic content. Blend PWHC was proportional to CMC content. In vitro procedures are useful in predicting the actions of fibre blends formulated to produce desirable effects in vivo.

Patulin surveillance in apple cider and juice marketed in Michigan.

Patulin is the most common mycotoxin found in apples and apple juices. The objective of this study was to determine the concentrations of patulin in (i) apple cider produced and marketed by Michigan apple cider mills during the fall seasons of 2002 to 2003 and 2003 to 2004 and (ii) apple juice and cider, including shelf-stable products, marketed in retail grocery stores in Michigan throughout 2005 and 2006. End product samples (n=493) obtained from 104 Michigan apple cider mills were analyzed for patulin concentration by using solid-phase extraction followed by high-performance liquid chromatography. Patulin was detected (> or =4 microg/liter) in 18.7% of all cider mill samples, with 11 samples (2.2%) having patulin concentrations of > or =50 microg/liter. A greater percentage of cider samples obtained from mills using thermal pasteurization contained detectable patulin (28.4%) than did those from mills using UV light radiation (13.5%) or no pathogen reduction treatment (17.0%). Among retail grocery store samples (n=159), 23% of apple juice and cider samples contained detectable patulin, with 18 samples (11.3%) having patulin concentrations of > or =50 microg/liter. The U.S. Food and Drug Administration (FDA) action level for patulin is 50 microg/kg. Some apple juice samples obtained from retail grocery stores had exceptionally high patulin concentrations, ranging up to 2700 microg/liter. Collectively, these results indicate that most apple cider and juice test samples from Michigan were below the FDA action level for patulin but that certain apple cider and juice processors have inadequate controls over patulin concentrations in final products. The industry, overall, should focus on improved quality of fruit used in juice production and improve culling procedures to reduce patulin concentrations.

Listeria in Raw Milk Soft Cheese: A Case Study of Risk Governance in the United States Using the IRGC Framework

Between 1980 and 1996 there were 30 known and reported outbreaks of foodborne illness associated with cheese consumption in the United States, Canada, Europe, and Scandinavia (Cody et al. 1999), and 16 of these outbreaks were associated with cheese produced using unpasteurised milk contaminated with one or more of the following pathogens— Brucella sp, Escherichia coli, Listeria monocytogenes, and Salmonella spp., and Yersinia enterocolitica (Teuber 2000). In this chapter, we will focus on only one of these pathogens—Listeria monocytogenes (Lm). Of the above outbreaks, three were caused by Lm, which resulted in 284 reported illnesses and 86 deaths (Teuber 2000). Periodic outbreaks of listeriosis from cheese have continued to occur; at least another six Lm outbreaks in the U.S., four in Europe, and two in Canada have been associated with cheese consumption since 1996 (de Valk et al. 2005; Food Safety Network 2005; Pagotto et al. 2006; U.S. Food and Drug Administration Center for Food Safety and Applied Nutrition [FDA CFSAN] et al. 2003).

High sucrose diets promote intestinal epithelial cell proliferation and tumorigenesis in APCMin mice by increasing insulin and IGF-I levels.

Epidemiological studies report that high sucrose consumption is associated with increased risk of colon cancer. One hypothesis is that this association is mediated by elevated circulatory insulin and IGF levels promoting intestinal proliferation. To test this hypothesis, APC Min mice and their wild type littermates were fed, starting at 4 wk of age, sucrose or cornstarch as the sole carbohydrate source in the absence or presence of low levels of dietary sulindac for 10 or 16 wk, respectively. APC Min mice fed sucrose had an increased tumor number in the proximal third of the small intestine in both studies and a higher incidence of papillary colon tumors in the 16-wk feeding study ( P ≤ 0.05). Mice fed sucrose (relative to cornstarch) had higher body weights and greater Ki67-labeling indexes in colonic epithelium than mice fed cornstarch in both feeding studies ( P ≤ 0.05). Furthermore, mice fed sucrose had higher serum glucose and liver IGF-I mRNA concentrations ( P ≤ 0.05) and tended to have higher serum insulin levels ( P = 0.08). These results support the hypothesis that high dietary sucrose intake promotes intestinal proliferation and tumorigenesis by increasing circulating levels of insulin and IGF-I.

Dietary carbohydrate source alters gene expression profile of intestinal epithelium in mice

High-sucrose consumption is associated with increased risk of human colon cancer. Our previous research indicated that high-sucrose diets (vs. cornstarch) promote intestinal epithelial cell proliferation and tumorigenesis as well as increase serum glucose and hepatic IGF-I mRNA levels in APC Min mice. To examine the role of functional pathways, in particular of IGF-I signaling, in sucrose-induced intestinal epithelial cell proliferation and tumorigenesis, we examined the effects of dietary carbohydrate source (sucrose vs. cornstarch) on gene expression in the intestinal epithelium using cDNA microarray and quantitative RT-PCR analysis. Dietary carbohydrate source significantly ( P < 0.05) altered mRNA expression of 109 known genes in the small intestinal epithelium, including many involved in metabolic pathways. Consumption of high-sucrose diets altered expression levels of genes involved in cell adhesion, cell cycle control, and transduction signaling, consistent with increased risk of intestinal tumorigenesis. High-sucrose intake also affected expression of genes involved in IGF-I signaling, including upregulating IGF-II and downregulating IGFBP3, which supports our hypothesis that IGF-I signaling could play a role in intestinal epithelial cell proliferation and tumorigenesis promoted by high-sucrose consumption.

Microbial levels in Michigan apple cider and their association with manufacturing practices

In recent decades, apple cider has been implicated in a series of outbreaks of foodborne illness. The objective of this study was to determine the presence and concentrations of pathogenic and indicator microorganisms in apple cider processed in Michigan and to evaluate the impact of thermal pasteurization, UV light radiation, and implementation of hazard analysis critical control point (HACCP) plans on these microbes. Cider samples were obtained from Michigan mills between 1997 and 2004 and analyzed for Escherichia coli O157:H7, Salmonella, generic E. coli, total coliforms, and aerobic bacteria. Neither E. coli O157:H7 nor Salmonella were detected in any tested cider samples, suggesting a very low frequency of pathogens in Michigan apple cider. The persistent and relatively high frequency of generic E. coli observed in samples obtained in all years indicates a continued risk of pathogen contamination in Michigan apple cider, especially when it is untreated. The use of thermal pasteurization or UV light radiation and reported implementation of HACCP plans were associated with lower frequency and counts of generic E. coli, total coliforms, and aerobic microorganisms. However, the relatively high counts of indicator organisms in some cider samples that were claimed to be treated according to these pathogen reduction measures indicates that some processors had inadequate practices, facilities, or equipment for pathogen reduction or did not consistently or adequately apply practices or pathogen-reduction equipment in an effective manner.