Functional foods and cardiovascular disease.

Functional foods are foods that, by virtue of physiologically active food components, provide health benefits beyond basic nutrition. Many functional foods have been found to be potentially beneficial in the prevention and treatment of cardiovascular disease, the leading cause of mortality in the United States. These foods include soybeans, oats, psyllium, flaxseed, garlic, tea, fish, grapes, nuts, and stanol- and sterol ester enhanced margarine. When eaten in adequate amounts on a consistent basis, these foods may aid in decreasing the risk of cardiovascular disease by several potential mechanisms: lowering blood lipid levels, improving arterial compliance, reducing low-density lipoprotein oxidation, decreasing plaque formation, scavenging free radicals, and inhibiting platelet aggregation.

The changing face of functional foods.

Consumers began to view food from a radically different vantage point in the 1990s. This 'changing face' of food has evolved into an exciting area of the food and nutrition sciences known as functional foods. Functional foods can be defined as those providing health benefits beyond basic nutrition and include whole, fortified, enriched or enhanced foods which have a potentially beneficial effect on health when consumed as part of a varied diet on a regular basis at effective levels. Interest in functional foods skyrocketed in the last decade due to a number of key factors, including the growing self-care movement, changes in food regulations and overwhelming scientific evidence highlighting the critical link between diet and health. The interest in functional foods has resulted in a number of new foods in the marketplace designed to address specific health concerns, particularly as regards chronic diseases of aging. In addition to new foods designed specifically to enhance health, however, functional foods can also include those traditional, familiar foods for which recent research findings have highlighted new health benefits or dispelled old dogma about potential adverse health effects. An excellent example is the American egg-Nature's original functional food. Eggs have not traditionally been regarded as a functional food, primarily due to concerns about their adverse effects on serum cholesterol levels. Furthermore, it is now known that there is little if any connection between dietary cholesterol and blood cholesterol levels and consuming up to one or more eggs per day does not adversely affect blood cholesterol levels. Finally, eggs are an excellent dietary source of many essential (e.g., protein, choline) and non-essential (e.g., lutein/zeaxanthin) components which may promote optimal health. Nutrition in the new millennium will be dramatically different than it was in the 20th century. Completion of the human genome project will facilitate the identification of humans predisposed to diet-related diseases. Targeted or 'prescription' nutrition will become the norm, enabling the food and medical industries to provide timely and individualized approaches to disease prevention and health promotion. The egg will continue to play an important role in the changing face of functional foods.

Effects of feeding 4 levels of soy protein for 3 and 6 wk on blood lipids and apolipoproteins in moderately hypercholesterolemic men.

BACKGROUND: Replacing animal protein with soy protein has been shown to reduce total and LDL-cholesterol concentrations in humans. However, the minimum amount of soy protein required for significant reduction of blood lipids is not known. OBJECTIVE: We evaluated the amount of soy protein needed to reduce blood lipids in moderately hypercholesterolemic men. DESIGN: Eighty-one men with moderate hypercholesterolemia (total cholesterol concentration between 5.70 and 7.70 mmol/L) were studied. After a 3-wk lead-in on a Step I diet, total cholesterol was measured and subjects were randomly divided into 5 groups. For 6 wk, each group received 50 g protein/d, which included isolated soy protein (ISP) and casein, respectively, in the following amounts: 50:0, 40:10, 30:20, 20:30, and 0:50 (control group) g. Blood was collected at baseline and weeks 3 and 6 of the intervention. RESULTS: At week 6, significant reductions (P < 0.05) from baseline compared with the control group were found for non-HDL and total cholesterol and apolipoprotein (apo) B for all ISP groups (except total cholesterol with 40 g ISP). At week 3, significant reductions (P < 0.05) were found in apo B for the groups that consumed >/=30 g ISP and in non-HDL cholesterol for the groups that consumed >/=40 g ISP. HDL-cholesterol, apo A-I, lipoprotein(a), and triacylglycerol concentrations were not significantly affected by dietary treatment. CONCLUSION: Our findings show that consuming as little as 20 g soy protein/d instead of animal protein for 6 wk reduces concentrations of non-HDL cholesterol and apo B by approximately 2.6% and 2.2%, respectively. 2000;71:-84.

Role of soy protein with normal or reduced isoflavone content in reversing bone loss induced by ovarian hormone deficiency in rats.

Soy protein, a rich source of isoflavones, fed immediately after an ovariectomy prevents bone loss in rats. Reports of the effectiveness of natural and synthetic isoflavones in preventing or treating osteoporosis led us to examine the effect of soy protein in reversing established bone loss. Seventy-two 95-d-old female Sprague-Dawley rats were assigned to 6 groups. The rats were either sham operated (SHAM; 2 groups) or ovariectomized (OVX; 4 groups) and then fed a casein-based, semipurified diet. Thirty-five days after surgery, 1 SHAM and 1 OVX group were killed to examine the occurrence of bone loss. Thereafter, the other SHAM and 1 OVX groups continued to receive the casein-based diet. Whereas the remaining 2 OVX groups received diets in which casein was replaced by soy protein with normal (OVX+SOY) or reduced (OVX+SOY-) isoflavone content for 65 days. The OVX control group had significantly lower femoral and fourth lumbar vertebral bone densities than the SHAM group. Femoral density of rats fed SOY or SOY- diets were not significantly different from SHAM or OVX controls. This suggests a slight reversal of cortical bone loss that may be partially due to higher femoral insulin-like growth factor I mRNA transcripts resulting from both the SOY and SOY- diets. The ovariectomy-induced increases in indexes of bone turnover were not ameliorated by either of the soy diets, suggesting that any positive effect of soy was achieved through enhanced bone formation rather than slowed bone resorption. Long-term consumption of soy or its isoflavones may be needed to produce small but continued increments in bone mass.

Bone-sparing effect of soy protein in ovarian hormone-deficient rats is related to its isoflavone content.

Our previous studies showed that a soy-protein diet prevents ovariectomy-induced bone loss. The purpose of this study was to determine whether isoflavones in soy protein are responsible for this bone-protective effect. Forty-eight 95-d-old Sprague-Dawley rats were divided into 4 groups: sham-operated fed a casein-based diet (SHAM), ovariectomized fed a casein-based diet (OVX+CASEIN), ovariectomized fed soy protein with normal isoflavone content (OVX+SOY), and ovariectomized fed soy protein with reduced isoflavone content (OVX+SOY-). The OVX+SOY group had significantly greater femoral bone density (in g/cm3 bone vol) than the OVX+CASEIN group, whereas OVX+SOY- was similar to OVX+CASEIN (mean +/- SD; SHAM, 1.522 +/- 0.041; OVX+CASEIN, 1.449 +/- 0.044; OVX+SOY, 1.497 +/- 0.030; OVX+SOY-, 1.452 +/- 0.030). Ovariectomy resulted in greater bone turnover as indicated by higher serum alkaline phosphatase activity, serum insulin-like growth factor I and insulin-like growth factor binding protein 3 concentrations, and urinary hydroxyproline. These increases were not affected by soy with either normal or reduced isoflavone content. Similarly, histomorphometry revealed a greater bone formation rate with ovariectomy, and this was not altered by the soy diets. The findings of this study suggest that isoflavones in soy protein are responsible for its bone-sparing effects. Further studies to evaluate the mechanism of action of isoflavones on bone are warranted.

Specific binding to protein kinase C by ingenol and its induction of biological responses.

We have examined the ability of ingenol to bind to and activate protein kinase C and to induce similar responses to the phorbol esters in biological systems. The rationale was that ingenol possesses the critical functionalities of the phorbol ester pharmacophore with the exception of the hydrophobic domain; it might therefore possess weak potency, although previous reports had indicated that ingenol was biologically inactive. Our data demonstrate that ingenol indeed binds to protein kinase C with a Ki of 30 microM and activates the enzyme. In addition, ingenol was biologically active in 3 separate cell systems, showing effects similar to the phorbol esters on morphological change, cell-cell communication, epidermal growth factor binding, arachidonic acid metabolite release, and ornithine decarboxylase activity. The 50% effective concentration values for the biological activity of ingenol were between 30 microM and 1 mM, varying somewhat with the cell system and type of response. The biological activity of ingenol in general supports the proposed models of the phorbol ester pharmacophore and imposes additional experimental constraints that the modeling must satisfy.

Incorporation of n-3 fatty acids into WB-F344 cell phospholipids inhibits gap junctional intercellular communication.

In this investigation, we demonstrate that rat liver epithelial (WB-F344) cells grown in medium supplemented with n-3 fatty acids (FA) results in the inhibition of gap junctional intercellular communication (GJIC). Cells incubated for 48 hr in medium containing 50 microM alpha-linolenate (18:3n-3) resulted in a 60% inhibition of GJIC, compared to control cells, while treatment with gamma-linolenate (18:3n-6) had no effect. Supplementation with octadecatetraenoate (18:4n-3), eicosapentaenoate (20:5n-3), and docosahexaenoate (22:6n-3), inhibited GJIC by 42%, 28%, and 18%, respectively. Incubation with each of the n-3 FA markedly increased the total n-3 FA content of cellular phospholipids (PL). Growing cells in medium containing 50 microM arachidonate (20:4n-6) plus 50 microM 18:3n-3 partially attenuated the inhibition of GJIC induced by 18:3n-3. The mechanism by which n-3 FA inhibit GJIC remains to be determined.

Inhibition of gap junction-mediated intercellular communication by alpha-linolenate.

The purpose of this investigation was to assess whether alterations in the fatty acid composition of rat liver epithelial (WB-F344) cell phospholipids would modulate gap junction-mediated intercellular communication (GJIC). WB-F344 cells were grown to confluency in culture medium supplemented with one of seven different fatty acids at a concentration of 50 microM for 48 h. Only alpha-linoleate (18:3 n-3) significantly inhibited GJIC. Saturated fatty acids (12:0, 16:0, and 18:0), a monounsaturated fatty acid (18:1 n-9), and n-6 polyunsaturated fatty acids (18:2 and 20:4) did not affect GJIC. The alpha-linolenate-induced inhibition of GJIC was not due to the activation of protein kinase C or intracellular hydroperoxide production, two lipid-dependent parameters previously shown to inhibit GJIC. In addition, alpha-linolenate did not alter membrane fluidity. Although the mechanism by which alpha-linolenate inhibits GJIC is unclear, changes in the fatty acid composition of cell phospholipids may be of critical importance. Subsequent to supplementation with alpha-linolenate, WB-F344 cell phospholipids had reduced 20:4 n-6 and elevated n-3 fatty acids. The results of this investigation emphasize the importance of current research into the influence of lipids on cell function and identify a new mechanism by which gap junctions can be modulated.

TPA-induced inhibition of gap junctional intercellular communication is not mediated through free radicals.

The present investigation was designed to determine whether the inhibition of gap junction-mediated intercellular communication (GJIC) induced by TPA (12-O-tetradecanoylphorbol-13-acetate) in rat liver epithelial (WB-F344) cells in vitro is mediated through free radical production. As assessed by fluorescence redistribution after photobleaching (FRAP) analysis, GJIC was significantly inhibited in cells treated for 1 hr with either 10 ng/ml TPA or 500 microM hydrogen peroxide (H2O2). Addition of 1000 U/ml catalase or 25 microM N',N'-diphenyl-p-phenylenediamine (DPPD) to TPA-treated cells did not alleviate the TPA-induced inhibition of GJIC. However, the concurrent addition of 1000 U/ml catalase to the culture medium prevented the H2O2 inhibition of GJIC. 2'-7'-dichlorofluorescein-mediated fluorescence, a measure of free radical production utilizing the Meridian ACAS 470 interactive laser cytometer, was not significantly increased in WB-F344 cells treated with 10 and 100 ng/ml TPA when compared to control cells. However, polymorphonuclear leukocytes (PMNs) treated for 10 min with 100 ng/ml TPA showed a substantial oxidative burst, as did WB-F344 cells treated for 1 hr with 500 microM H2O2. The concurrent addition of 1000 U/ml catalase to the culture medium attenuated H2O2-mediated free radical production in both PMNs and WB-F344 cells. Data from this study do not support a role for free radicals in the TPA-induced inhibition of GJIC in WB-F344 cells.

Exercise attenuates diet-induced arteriosclerosis in the adult rat.

The present investigation was conducted to assess the effects of exercise on diet-induced arteriosclerosis in retired breeder rats. Thirty-two 8- to 9-mo-old male Sprague-Dawley rats were allocated to one of four treatment groups: hypercholesterolemic diet-exercise (HE), hypercholesterolemic diet-sedentary (HS), normocholesterolemic diet-exercise (NE) and normocholesterolemic diet-sedentary (NS). The hypercholesterolemic diet contained 10% lard and 0.4% cholesterol. Exercise consisted of running on a motor-driven treadmill at an 8% grade 1 h per day at 0.5 mph, 6 d weekly for 5 mo. A histological assessment of the aortas demonstrated that although grossly visible atherosclerotic plaques were absent, there were significant microscopic differences among groups in the thoracic aorta, iliac bifurcation and aortic arch. Aortic histopathological changes were greatest in cross sections from rats in the HS group. The accumulation of collagen and sulfated mucosubstances was greater in the HS versus HE group. Our results demonstrate a beneficial effect of exercise on diet-induced histopathological changes in the aortas of male retired breeder rats.

The effect of exercise and hyperlipemic diet on aortic histopathology in the rat.

Histological changes in the aorta were used as criteria to assess the effect of treadmill exercise on the development of atherosclerosis in rats consuming a moderately hyperlipemic diet. While grossly visible atherosclerotic plaques were absent, microscopic examination of sections of the abdominal aorta were distinctly different between exercised and sedentary rats. Aortas of sedentary rats fed a diet containing lard and cholesterol had a high degree of plaque development, fat accumulation, mineralization, erosion and necrosis. Aortas of exercised rats fed the same diet had considerably less atherogenic involvement. Both hyperlipemic groups had greater plaque formation than sedentary rats fed a normolipemic diet. In addition, there were marked differences in plasma lipids; exercise ameliorated the diet-induced changes in plasmatic atherogenic lipids. Our results indicate that exercise retards the development of atherosclerosis in rats fed a hyperlipemic diet.