Effects of various fiber additions on lipid digestion during in vitro digestion of beef patties.

The purpose of this study was to examine the effect of various fiber additions on lipid digestion during the in vitro digestion of beef patties. The control patties were prepared with 90.5% lean meat and 9.5% tallow. Treatments consisted of 90% lean meat with 9.5% tallow and either 0.5% cellulose, 0.5% chitosan, or 0.5% pectin. The beef patties were then passed through an in vitro digestion model that simulated the composition of the mouth, stomach, and small intestine juices. The change in structure and properties of the lipid droplets was monitored by laser scanning confocal fluorescence microscopy. In general, there was a decrease in lipid droplet diameter as the droplets moved from mouth to stomach to small intestine. The amount of free fatty acid dramatically increased after in vitro digestion in all beef patties. The amount of free fatty acid was, however, lower in beef patties containing chitosan and pectin than other beef patties after in vitro digestion. Beef patties containing various fibers had lower thiobarbituric acid-reactive substances (TBARS) values than samples with no fibers. Among the samples to which fibers were added, chitosan and pectin had lower TBARS than beef patties with cellulose. The cholesterol content decreased after in vitro digestion in all beef patties but was not different among the beef patties before and after in vitro digestion. These results enhance our understanding of the physicochemical and structural changes that occur to ground beef within the gastrointestinal tract.

Comparative effect of seeds of Rhynchosia volubilis and soybean on MG-63 human osteoblastic cell proliferation and estrogenicity.

The seeds of Rhynchosia volubilis (SRV) (Leguminosae) and soybean have been used in oriental folk medicine to prevent postmenopausal osteoporosis. Their beneficial effects are caused by a high content of isoflavone, which function as partial agonists or antagonists of estrogen. To compare the estrogenic effects of SRV and soybean on the MG-63 osteoblastic cell proliferation, 70% methanol extracts of SRV or soybean were treated on MG-63 cells. Although biphasic over a concentration range of 0.001 mg/ml-0.1 mg/ml, both SRV and soybean extracts increased MG-63 cell proliferation. However SRV was more effective at increasing the cell proliferation that paralleled with the greater estrogenic effects as determined by estrogen receptor alpha (ERalpha) expression, an estrogenic response element (ERE)-luciferase activity and the selective expression of insulin-like growth factor-I (IGF-I). SRV-induced IGF-I expression resulted from increases in the mRNA levels. Despite the increased expression of ERbeta, ERE activity and IGF-I expression by soybean were lower than those by SRV. Furthermore, the comparable estrogenic effects between SRV and the combined treatment of genistein and daidzein standards at 0.5 x 10(-8) M, which is a concentration of these two isoflavones similar to that of SRV at 0.001 mg/ml, demonstrate that the greater estrogenicity of SRV for MG-63 cell proliferation is mediated by the synergism of low levels of isoflavones for the selective expression of IGF-I.

Effect of dietary restriction on age-related increase of liver susceptibility to peroxidation in rats.

Dietary restriction (DR) increases life span and decreases age-related diseases in experimental animals. It has received a great deal of attention in connection with the relationship between aging, nutrition, and oxidative stress because oxidative injury in several organ systems is a prominent feature in aging. We investigated the possibility that DR can protect vulnerable liver lipids against age-related increases of peroxidation. Male Fischer 344 rats fed ad libitum (AL) or dietarily restricted (maintained on 60% of AL food intake) were killed by decapitation at 4 (young) or 12 mon (adult) of age. Phosphatidylcholine hydroperoxide (PCOOH) concentration of liver was determined using a chemiluminescent high-performance liquid chromatographic method. Liver PCOOH increased with age in adult rats, but less of an increase of PCOOH was seen in DR rats, which is consistent with results on production of thiobarbituric acid-reactive substances and oxygen-derived free radicals. No significant differences were found in liver superoxide dismutase and catalase activity between AL and DR groups of young and adult rats. Liver triglyceride and cholesterol contents were lower in DR than AL rats at 12 mon. Fatty acid compositions of phosphatidylcholine and phosphatidylethanolamine indicated that the ratio of (20:3n-6 + 20:4n-6)/18:2n-6, an index of linoleic acid (18:2n-6) desaturation, was lower in DR than in AL rats. We concluded that DR suppresses age-related oxidative damage in liver by modulating the amount of lipid as well as fatty acid composition.

Dietary (n-6) and (n-3) fatty acids and energy restriction modulate mesenteric lymph node lymphocyte function in autoimmune-prone (NZB x NZW)F1 mice.

We previously showed that dietary fish oil (FO) and energy restriction (R) have beneficial anti-inflammatory properties in the peripheral blood and spleens of (NZB x NZW)F1 (B/W) lupus-prone mice. Furthermore, unsaturated fatty acids also were shown in the past to influence mesenteric lymph node (MLN) lymphocyte function in healthy young rats. The MLN play a pivotal role in mediating food allergy. To date, the effect of R on intestinal immunity is not well understood; therefore we determined the effect of diet on MLN lymphocyte function. Mice were given either free access to a 5 g/100 g corn oil (CO) or fish oil (FO) diet or the same corn oil (CR) or fish oil (FR) diets restricted to 60% of the intake of the control group. At the age of 4 (young) and 8 (old) mo, MLN lymphocytes were isolated and B- (CD19(+)) and T-lymphocyte subsets (CD4(+) and CD8(+)) were determined by flow cytometry. Additional MLN lymphocytes were placed in culture with or without concanavalin A and culture supernatants collected after 72 h for cytokine and immunoglobulin (Ig) quantitation by ELISA. Aging significantly (P < 0.05) decreased both CD4(+) and CD8(+) T-lymphocytes. Spontaneous and activation-induced interleukin-4 (IL-4), IL-10, and interferon-gamma secretion were greater while IL-2 was lower in CO-fed old mice compared to CO-fed young mice. In contrast, CR or FO alone partially blunted the age-dependent alterations in T-lymphocyte ratios including cytokine and Ig secretion, whereas the FR diet significantly (P < 0.005) normalized the accelerated aging effects on these immune variables. We show for the first time that FR is a far more potent anti-inflammatory therapy than either CR or FO alone in modulating MLN lymphocyte function.

Application of flow injection--chemiluminescence to the study of radical scavenging activity in plants.

Chemiluminescence (CL) was observed during the oxidation of luminol (2 mg/L). mediated by 0.06% hydrogen peroxide (H(2)O(2)) and cytochrome c (10 mg/L). CL intensity was decreased by the presence of radical scavengers and the reduction was linearly proportional to the concentration and ability of scavengers; butylated hydroxytoluene (BHT), caffeic acid and gallic acid. The order of effectiveness as radical scavengers was gallic acid > caffeic acid > BHT, which shows that the number of hydroxyl groups (OH) in the B-ring of flavonoids plays a key role in a good radical scavenging activity. Of eight catechins obtained from green tea extracts, (-)-catechin was the least effective and (-)-epigallocatechin gallate (EGCg) showed the strongest activity. This result indicates that the stereoscopic structure between the C-3 group and the B ring of flavonoids as well as substituents at the C-3 position make a contribution to radical scavenging activity. Of the tested Chinese herbal ingredients, five species of ingredients represented more than 90% of the radical scavenging activity.

The antioxidative effect of ganhuangenin against lipid peroxidation.

The antioxidative effect of ganhuangenin (GHG), isolated from Scutellaria baicalensis Georgi, was examined by measuring its ability to suppress the formation of phospatidylcholine hydroperoxide (PCOOH). The results show that a pretreatment with GHG effectively suppressed PCOOH formation, which was initiated by the peroxyl-generating oxidant, AAPH (2,2'-azobis-2-aminopropane hydrochloride). The protective action of GHG against the formation of the PCOOH was observed in liver, lung, and kidney. When compared with other known antioxidants, we found the antioxidative potency of GHG to be greater than that of alpha-tocopherol. Our data strongly indicate that GHG is a powerful antioxidant against lipid peroxidation and is, therefore, responsible for this prophylactic effect.

Effect of unsaturated fatty acids and alpha-tocopherol on immunoglobulin levels in culture medium of rat mesenteric lymph node and spleen lymphocytes.

Mesenteric lymph node (MLN) and spleen lymphocytes of Sprague-Dawley rats were cultured with 1 mM unsaturated fatty acids (UFAs) with or without 100 microM alpha-tocopherol (Toc), and the immunoglobulin content and thiobarbituric acid (TBA) value of the culture media were measured to clarify the relationship between lipid peroxidation and the IgE level in the culture medium. The increase in the IgE content and TBA value induced by UFAs was alleviated in the presence of Toc in both lymphocytes, and was correlated well with their oxidation rates in most cases. Gamma-linolenic acid enhanced the IgE level much more than would be expected from its oxidation rate in both lymphocytes, and linoleic acid showed similarly high activity only in splenocytes. These results suggest that lipid peroxidation is partly responsible for the enhancement of IgE level induced by UFAs.

Dietary fibers modulate indices of intestinal immune function in rats.

To explore the immunological roles of dietary fiber, male 4-wk-old Sprague-Dawley rats were fed for 2 wk cellulose (water-insoluble), konjak mannan (water-soluble), pectin (water-soluble) or chitosan (acid-soluble) at 5 g/100 g diet. Serum IgE concentrations in rats fed konjak mannan, pectin and chitosan were significantly lower than in those fed cellulose (mean +/- SEM: 5.0 +/- 1.1, 3.6 +/- 1.3, 3.0 +/- 1.2 and 9.6 +/- 1.9 microg/L, respectively). Rats fed pectin had significantly higher serum IgA and IgG concentrations (358 +/- 38 and 424 +/- 36 mg/L for IgA and IgG, respectively) than those fed cellulose (240 +/- 31 and 337 +/- 25 mg/L) or chitosan (176 +/- 22 and 379 +/- 23 mg/L), while the IgM concentration did not differ among the groups. Concentrations of IgA, IgG and IgM in mesenteric lymph node (MLN) lymphocytes generally were greater, while IgE concentration was lower, in rats fed pectin and chitosan than in those fed cellulose. The proportion of CD4+ T-cells in MLN lymphocytes was also dietary fiber-dependent, and the CD4+/CD8+ ratio was significantly higher in the pectin fed group than in all other groups. Under certain experimental conditions, MLN lymphocytes from rats fed pectin had markedly greater interferon-gamma concentration than cells from other groups, while the effect on tumor necrosis factor-alpha concentration was less marked. Thus, dietary fiber may have an immunoregulatory effect on the intestinal immune system of rats.

Stimulating effect of xanthene dyes on immunoglobulin produced in vitro by rat spleen lymphocytes.

The effects of food additives on immunoglobulin produced in rat splenic lymphocytes were examined. The xanthene dye, Rose Bengal, enhanced IgE production, while inhibiting the production of IgG and IgM, at 50 microM. Among the xanthene dyes, Rose Bengal having 4 iodine and 4 chlorine atoms exerted the highest Ig production-regulating activity in splenocytes, and dihalogenated fluorescein, a diiodo compound, exerted similar activity, while the dichloro and dibromo compounds did not. These results suggest that halogen atoms, especially the iodine atom, in xanthene dyes play an important role in regulation of Ig production.

Effects of gamma-linolenic acid and its positional isomer pinolenic acid on immune parameters of brown-Norway rats.

Male Brown-Norway rats given purified diets containing safflower oil (SFO, linoleic acid, 18:2 n-6), evening primrose oil (EPO, gamma-linolenic acid, 6,9,12- 18:3 n-6) or Korean pine seed oil (PSO, 5,9,12- 18:3) at the 10% level were immunized twice with intraperitoneal ovalbumin, on days 14 and 35 of the feeding diets, and killed one day after the second booster. The relative population of CD4+ T-lymphocytes in the spleen was significantly lower in rats fed SFO than in those fed EPO or PSO, while that of CD8+ subsets remained unchanged. There was a significant increase in the splenic production of IgG and IgE in the PSO group compared to the SFO group, while EPO significantly increased IgE. The periodical response patterns of the serum levels of IgG and IgE varied depending on the source of dietary fats, and the initial rise of total immunoglobulins tended to be higher in the EPO group. The release by peritoneal exudate cells of histamine was comparable among three groups irrespective of saturation by calcium ionophore A23187, while PSO significantly increased leukotriene B4 production. These observations not only indicate specific roles of gamma-linolenic acid but also diverse influences of different octadecatrienoic acids in various immune measurements.

Effect of unsaturated fatty acids and antioxidants on immunoglobulin production by mesenteric lymph node lymphocytes of Sprague-Dawley rats.

The effect of UFA on Ig production by rat MLN lymphocytes was examined to clarify their influence on allergic reactions. A 4-h treatment at 1 mM inhibited the production of IgA, IgG, and IgM by the lymphocytes, but stimulated IgE production. The IgE production-stimulating activity became stronger with increasing number of carbon atoms and/or double bonds. However, no saturated fatty acid with carbon numbers from 12 to 18 affected IgE production by the lymphocytes. Hydrogen peroxide exerted Ig production-regulating activity similar to that of UFA, suggesting that the effect of UFA is at least partly due to oxidation products. Thus, the effect of antioxidants on the Ig production-regulating activity of arachidonic acid was examined. alpha-Tocopherol and BHT annulled the stimulation of IgE production by arachidonic acid, but ascorbic acid was not effective. The IgE production-enhancing activity of UFA was closely related to their oxidation rate in culture medium. These results suggest that UFA enhance the allergic reaction through the stimulation of IgE production and the inhibition of IgA production, and that hydrophobic antioxidants are partially effective to annul the adverse effect of UFA.

Effects of n-3 polyunsaturated fatty acids and lectins on immunoglobulin production by spleen lymphocytes of Sprague-Dawley rats.

We examined the effects of n-3 polyunsaturated fatty acid (PUFA), such as alpha-linolenic (alpha-LA), eicosapentaenoic (EPA), and docosahexaenoic acid (DHA) on immunoglobulin (Ig) production by spleen lymphocytes of Sprague-Dawley rats. n-3 polyunsaturated fatty acid (PUFA) strongly inhibited the production of IgA and IgM and that of IgG weakly at 100 microM. When the lymphocytes were treated with n-3 PUFA in the presence of other inhibitory biomaterials such as lectins, some PUFA attenuated their inhibitory effect on Ig production. In the presence of concanavalin A (ConA), all n-3 PUFA attenuated the inhibitory effect of ConA on the production of IgM or IgG but increased its inhibition of IgA synthesis. Thus, the interaction of n-3 polyunsaturated fatty acid and lectins in spleen interfere with each other or the expression of Ig production regulating activity.

Effects of sesamin and alpha-tocopherol, individually or in combination, on the polyunsaturated fatty acid metabolism, chemical mediator production, and immunoglobulin levels in Sprague-Dawley rats.

Feeding sesamin and alpha-tocopherol in combination, both at the 0.5% dietary level, to Sprague-Dawley rats for 3 weeks resulted in a trend toward decreasing the proportion of 20:4n-6 and 22:5n-6 and increasing that of 18:2n-6 in phosphatidylcholine from various tissues, suggesting interference with the metabolism of linoleic acid. This dietary manipulation significantly reduced the production of leukotriene C4 in the lung, the splenic production of leukotriene B4, and reduction of the plasma histamine level. Simultaneous administration of sesamin and alpha-tocopherol significantly increased the production of IgA, IgG, and IgM by mesenteric lymph node lymphocytes, while the IgE level tended to be reduced. These effects were not necessarily apparent by feeding these compounds separately. Thus, sesamin and alpha-tocopherol in combination would be effective for regulating the eicosanoid production and modifying the immune function.

Free bile acids inhibit IgE production by mouse spleen lymphocytes stimulated by lipopolysaccharide and interleukins.

The interaction of IL-4, IL-5, and free bile acids with the immunoglobulin production by mouse spleen lymphocytes was studied to examine their immunoregulatory activity. In the absence of lipopolysaccharide (LPS), IL-4 enhanced the IgE and IgG production significantly and the IgA production weakly, but not the IgM production. On the other hand, IL-5 had an inhibitory tendency on the IgE and IgA production, though not significantly. In the presence of LPS, both IL-4 and IL-5 significantly enhanced the IgE production by mouse splenic lymphocytes. When the lymphocytes were cultured with the physiological concentration of free bile acids (10 microM) and LPS for 3 days, chenodeoxycholic acid inhibited the IgE production, but cholic and deoxycholic acids did not. In the presence of IL-4 or IL-5, these bile acids cancelled the stimulatory effects of interleukins and rather significantly inhibited the IgE production. These results suggest that these free bile acids act as an anti-allergic agent.

Effects of bile acids and lectins on immunoglobulin production in rat mesenteric lymph node lymphocytes.

We investigated the effect of bile acids either alone or in combination with lectins on immunoglobulin (Ig) production in vitro of rat mesenteric lymph node (MLN) lymphocytes to examine their immunoregulatory activities. Among free bile acids examined, chenodeoxycholic acid stimulated IgE production by MLN lymphocytes and inhibited IgA production at the concentration of 0.3 mM, whereas cholic and deoxycholic acids exerted the comparable effect at 3 mM. Among conjugated bile acids, deoxycholic acid derivatives stimulated IgE production more strongly than cholic acid derivatives. On the other hand, free and conjugated bile acids did not affect IgG production. The IgE production by MLN lymphocytes was stimulated by concanavalin A and inhibited by pokeweed mitogen, and the effect of phytohemmagglutinin and lipopolysaccharide was marginal. These lectins did not affect IgA and IgG production by the lymphocytes. In the presence of lectins, free bile acids affected IgE production at 0.03 mM. These results suggest the possibility that bile acid is a stimulant for food allergy.