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Bovine milk antibodies for health.

The immunoglobulins of bovine colostrum provide the major antimicrobial protection against microbial infections and confer a passive immunity to the newborn calf until its own immune system matures. The concentration in colostrum of specific antibodies against pathogens can be raised by immunising cows with these pathogens or their antigens. Immune milk products are preparations made of such hyperimmune colostrum or antibodies enriched from it. These preparations can be used to give effective specific protection against different enteric diseases in calves and suckling pigs. Colostral immunoglobulin supplements designed for farm animals are commercially available in many countries. Also, some immune milk products containing specific antibodies against certain pathogens have been launched on the market. A number of clinical studies are currently in progress to evaluate the efficacy of immune milks in the prevention and treatment of various human infections, including those caused by antibiotic resistant bacteria. Bovine colostrum-based immune milk products have proven effective in prophylaxis against various infectious diseases in humans. Good results have been obtained with products targeted against rotavirus, Shigella flexneri, Escherichia coli, Clostridium difficile, Streptococcus mutans, Cryptosporidium parvum and Helicobacter pylori. Some successful attempts have been made to use immune milk in balancing gastrointestinal microbial flora. Immune milk products are promising examples of health-promoting functional foods, or nutraceuticals. This review summarises the recent progress in the development of these products and evaluates their potential as dietary supplements and in clinical nutrition.

Adult↗

Yak milk casein as a functional ingredient: preparation and identification of angiotensin-I-converting enzyme inhibitory peptides.

Yak milk casein derived from Qula, a traditional Tibetan acid curd cheese, was hydrolyzed by six commercially available proteases (Trypsin, Pepsin, Alcalase, Flavourzyme, Papain and Neutrase). These hydrolysates were assayed for their inhibitory activity of Angiotensin-I-converting enzyme (ACE). The hydrolysates obtained by Neutrase from Bacillus amyloliquefaciens showed the highest ACE inhibitory activity. The IC50 value of Neutrase-hydrolysate was 0.38 mg/ml. The hydrolysate obtained by Neutrase was further separated by consecutive ultra-filtration with 10 kDa and then with 6 kDa molecular weight cut-offs into different permeated parts and fractionated by gel filtration chromatography with a Sephadex G-25 column. The active fraction was subjected to RP-HPLC, in which five peaks were purified and identified. Amino acid sequence analysis confirmed that the peptides and origins were as follows: YQKFPQY (alphas2-CN; f89-95), LPQNIPPL (beta-CN; f70-77), SKVLPVPQK (beta-CN; f168-176), LPYPYY (kappa-CN; f56-61) and FLPYPYY (kappa-CN; f55-61). Their amino acid sequences matched well with those of known bioactive peptides from bovine casein. The results indicated that yak milk casein could be a resource to generate antihypertensive peptides and be used as multifunctional active ingredients for many value-added functional foods as well as a traditional food protein.

Amino Acid Sequence↗

Phytosterols in sea buckthorn (Hippophaë rhamnoides L.) berries: identification and effects of different origins and harvesting times.

Sterols in seeds, pulp/peel fractions, and whole berries of sea buckthorn (Hippophaë rhamnoides L.) samples belonging to two major subspecies (sinensis and rhamnoides) from Finland and China were analyzed as TMS derivatives by gas chromatography-mass spectrometry after saponification of the oils. The total sterol contents in the seeds, the fresh pulp/peel, and the whole berries were 1200-1800, 240-400, and 340-520 mg/kg, respectively. The corresponding values in the extracted oils were 12-23, 10-29, and 13-33 g/kg. Sitosterol constituted 57-76 and 61-83%, respectively, of the seed and pulp/peel sterols. The sterol content and composition showed little variation between subspecies and collection sites. Different harvesting dates showed significant effects on the levels of some sterols both in the seeds and in the pulp/peel. The sterol profiles obtained are useful for characterizing sea buckthorn and detecting adulterations of the valuable oils. The information provided by the present investigation is also important for further chemical investigation of sea buckthorn sterols and industrial utilization of the berries as a raw material of functional foods.

Mass Spectrometry↗

Antioxidant activity of polyphenols in carob pods.

We extracted polyphenols from carob (Ceratonia siliqua L.) pods, and evaluated the in vitro antioxidant activity of the crude polyphenol fraction (CPP). The total polyphenol content in CPP determined by the Folin-Ciocalteu method was 19.2%. The condensed tannin content determined by the vanillin and proanthocyanidin assay systems was 4.37% and 1.36%, respectively. beta-Carotene bleaching, 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging, inhibition of lipid peroxidation by the erythrocyte ghost, and microsomal assay systems were used to evaluate the antioxidant activity. CPP showed a stronger inhibitory effect against the discoloration of beta-carotene than other polyphenol compounds such as catechins and procyanidins. CPP had weaker antioxidant activity in the DPPH free radical scavenging, the erythrocyte ghost, and microsomal systems than authentic polyphenol compounds at the same concentrations. The activity adjusted by the polyphenol concentration was, however, comparable to that of authentic polyphenol compounds. Considering most carob pods are discarded and not effectively utilized at present, these results suggested that carob pods could be utilized as a functional food or food ingredient.

Antioxidants↗

Comparison of different egg albumen fractions as sources of ovomucin.

Ovomucin was fractionated from whole egg albumen, thick egg albumen, liquid egg albumen, and a liquid egg albumen filtration byproduct by using the isoelectric precipitation method. The amounts of ovomucin measured in the above-mentioned fractions were 280, 340, 500, and 520 mg per 100 g of albumen, respectively. There was great variation between the beta-ovomucin contents of the different albumen fractions. Whole egg albumen contained about 25 mg of beta-ovomucin in 100 g of albumen, whereas thick egg albumen, liquid egg albumen, and the filtration byproduct contained about 1.5, 3, and 5 times more beta-ovomucin, respectively, as compared to whole egg albumen. The results indicate that both the liquid egg albumen fraction and especially the filtration byproduct fraction appear to be potential sources of ovomucin when it is used as an ingredient for functional foods.

Carbohydrates↗

Effects of management on the yield and high-molecular-weight polysaccharide content of shiitake (Lentinula edodes) mushrooms.

Shiitake (Lentinula edodes (Berk.) Pegler) mushroom production in the United States has increased greatly over the last twenty years. Additional expansion of the shiitake mushroom market should be possible if the product can be marketed as a functional food, i.e., a food that has health-promoting effects beyond its nutritional value. High-molecular-weight polysaccharides (HMWP) including lentinan in shiitake may promote human health. This study was conducted to determine if management protocols influence the HMWP of shiitake mushrooms. Results indicate that measuring the total carbohydrate content of water-extractable, ethanol-insoluble polysaccharides was a simple way to estimate HMWP. Results also indicate that log-grown shiitake contained more HMWP than did substrate-grown shiitake. Among log-grown shiitake, both mushroom strain and tree species influenced HMWP content. The results suggest that there is considerable variation among shiitake mushrooms in HMWP content and that production protocols influenced the HMWP content of mushrooms.

Agriculture↗

Effects of different origins and harvesting time on vitamin C, tocopherols, and tocotrienols in sea buckthorn (Hippophaë rhamnoides) berries.

Vitamin C, tocopherols, and tocotrienols in berries of wild and cultivated sea buckthorn (Hippophaë rhamnoides L.) of different origins and harvesting dates were determined with HPLC. Wild berries of subsp. sinensis, native to China, contained 5-10 times more vitamin C in the juice fraction than the berries of subsp. rhamnoides from Europe and subsp. mongolica from Russia (4-13 vs 0.02-2 g/L juice). Genetic background and berry-harvesting date were two primary factors determining the vitamin C content in the berries. Crossing different subspecies influenced the vitamin C content to some extent. For bushes cultivated in southwest Finland, the best berry-harvesting date for high vitamin C content was the end of August. The seeds of subsp. sinensis contained less tocopherols and tocotrienols (average 130 mg/kg) compared with seeds of subsp. rhamnoides (average 290 mg/kg) and mongolica (average 250 mg/kg). The fruit flesh of sinensis berries had contents of tocopherols and tocotrienols 2-3 times higher than those found in the other two subspecies (120 mg/kg vs 40 mg/kg in rhamnoides and 50 mg/kg in mongolica). The fresh whole berries of subsp. sinensis were clearly the best source of total tocopherols and tocotrienols. The total content of tocopherols and tocotrienols in the soft parts of the berries reached the maximum level around early- to mid-September, whereas the content in seeds continued to increase until the end of November. The excellent combination of the highest content of vitamin C and tocopherols and tocotrienols makes the berries of subsp. sinensis an optimal raw material for nutritional investigation as a candidate for functional foods with special antioxidative properties.

Agriculture↗

Oil goldenberry (Physalis peruviana L.).

Whole berries, seeds, and pulp/peel of goldenberry (Physalis peruviana L.) were compared in terms of fatty acids, lipid classes, triacylglyerols, phytosterols, fat-soluble vitamins, and beta-carotene. The total lipid contents in the whole berries, seeds, and seedless parts were 2.0, 1.8, and 0.2% (on a fresh weight basis), respectively. Linoleic acid was the dominating fatty acid followed by oleic acid as the second major fatty acid. Palmitic and stearic acids were the major saturates. In pulp/peel oil, the fatty acid profile was characterized by higher amounts of saturates, monoenes, and trienes than in whole berry and seed oils. Neutral lipids comprised >95% of total lipids in whole berry oil and seed oil, while neutral lipids separated in lower level in pulp/peel oil. Triacylglycerols were the predominant neutral lipid subclass and constituted ca. 81.6, 86.6, and 65.1% of total neutral lipids in whole berry, seed, and pulp/peel oils, respectively. Nine triacylglycerol molecular species were detected, wherein three species, C54:3, C52:2, and C54:6, were presented to the extent of approximately 91% or above. The highest level of phytosterols was estimated in pulp/peel oil that contained the highest level of unsaponifiables. In both whole berry and seed oils, campesterol and beta-sitosterol were the sterol markers, whereas Delta5-avenasterol and campesterol were the main 4-desmethylsterols in pulp/peel oil. The tocopherols level was much higher in pulp/peel oil than in whole berry and seed oils. beta- and gamma-tocopherols were the major components in whole berry and seed oils, whereas gamma- and alpha-tocopherols were the main constituents in pulp/peel oil. beta-Carotene and vitamin K(1) were also measured in markedly high levels in pulp/peel oil followed by whole berry oil and seed oil, respectively. Information provided by the present work is of importance for further chemical investigation of goldenberry oil and industrial utilization of the berries as a raw material of oils and functional foods.

Fatty Acids↗

Stability and emulsion-forming ability of water-soluble fish myofibrillar protein prepared by conjugation with alginate oligosaccharide.

Carp myofibrillar protein (Mf) was conjugated with alginate oligosaccharide (AO) through the Maillard reaction under low relative humidity, and the functional properties of the Mf-AO conjugate were investigated under different NaCl concentrations and pH levels. Mf became highly solubilized at lower NaCl concentrations by conjugation with AO, with a slight loss of available lysine. The thermal stability of Mf was effectively improved by conjugation with AO. Heat treatment at 80 degrees C for 2 h had no effect on the solubility of the Mf-AO conjugate attached to 227 microg/mg of AO regardless of the NaCl concentration and pH. Furthermore, the Mf-AO conjugate showed excellent emulsion-forming ability regardless of NaCl concentration. The improved functionalities of Mf by conjugation with AO remained even at a nearly isoelectric point. These results indicate that conjugation with AO through the Maillard reaction is an effective way to prepare high-functional food material from fish muscle protein.

Alginates↗

Effect of proteolytic squid protein hydrolysate on the state of water and dehydration-induced denaturation of lizard fish myofibrillar protein.

With the goal of preparing low-cost functional food, squid protein hydrolysate (SPH) was extracted from four squid species by protease treatment. Peptides are the major components (approximately 84-88%) of the SPH. The stabilization effects of 5% SPH (dried weight/wet weight) on the state of water and the denaturation of frozen lizard fish Saurida wanieso myofibrillar protein (Mf) were evaluated on the basis of desorption isotherm curves with respect to Ca2+-ATPase inactivation and the presence of unfrozen water, which was determined using differential scanning calorimetry during dehydration, and the effects were compared with those of sodium glutamate. The Mf with SPH was found to contain higher levels of monolayer and multilayer sorption water, resulting in decreased water activity and Ca2+-ATPase inactivation. The amount of unfrozen water in Mf with SPH increased significantly, suggesting that the peptides of SPH stabilized water molecules on the hydration sphere of Mf, which maintained the structural stability of Mf, and therefore suppressed dehydration-induced denaturation. The effect by SPH was less than that by sodium glutamate.

Animals↗

Identification of flavonol and xanthone glycosides from mango (Mangifera indica L. Cv. "Tommy Atkins") peels by high-performance liquid chromatography-electrospray ionization mass spectrometry.

Flavonol O- and xanthone C-glycosides were extracted from mango (Mangifera indica L. cv. "Tommy Atkins") peels and characterized by high-performance liquid chromatography-electrospray ionization mass spectrometry. Among the fourteen compounds analyzed, seven quercetin O-glycosides, one kaempferol O-glycoside, and four xanthone C-glycosides were found. On the basis of their fragmentation pattern, the latter were identified as mangiferin and isomangiferin and their respective galloyl derivatives. A flavonol hexoside with m/z 477 was tentatively identified as a rhamnetin glycoside, which to the best of our knowledge, has not yet been reported in mango peels. The results obtained in the present study confirm that peels originating from mango fruit processing are a promising source of phenolic compounds that might be recovered and used as natural antioxidants or functional food ingredients.

Chromatography, High Pressure Liquid↗

Stoichiometric and kinetic studies of phenolic antioxidants from Andean purple corn and red-fleshed sweetpotato.

Stoichiometric and kinetic values of phenolics against DPPH (2,2-diphenyl-1-picrylhydrazyl) were determined for Andean purple corn (Zea mays L.) and red sweetpotato (Ipomoea batatas). Both crops had higher antioxidant capacity and antiradical kinetics than blueberries and higher or similar anthocyanin and phenolic contents. The second-order rate constant (k(2)) was 1.56, 1.12, 0.57, and 0.26 (mg antiradical/mL)(-1) s(-1) for red sweetpotato, Trolox, purple corn, and blueberry, respectively. On the molar basis of active hydroxyl groups, k(2)' showed the same order as for k(2). Corn cob and sweetpotato endodermis contributed the most in phenolic compounds and antioxidant capacity. Both crops studied can be considered as excellent novel sources of natural antioxidants for the functional food and dietary supplement markets.

Anthocyanins↗

Evaluation of a commercial enzyme-based serum cholesterol test kit for analysis of phytosterol and phytostanol products.

Plant sterols (phytosterols) have been shown to possess serum cholesterol-lowering properties. In recent years, several phytosterol-enriched functional food products have been developed and marketed. Some phytosterol products contain common unsaturated sterols and some contain a subset of phytosterols called phytostanols (saturated sterols, also called plant stanols). Current methods for the quantitative analysis of plant sterols are labor intensive and require sophisticated gas or liquid chromatographs. In this study, a popular commercial spectrophotometric serum cholesterol test kit was evaluated for the analysis of plant sterols. The results indicate that the method could be modified to analyze phytosterols and phytostanols by increasing the incubation time. Both free phytosterols and fatty acyl phytosteryl esters were quantitatively analyzed, but ferulate phytosteryl esters, such as those that are found in corn and other cereals, were not hydrolyzed by the enzymes in the test kit and therefore were not detected.

Cholesterol↗

Quantification of the interactions between beta-lactoglobulin and pectin through capillary electrophoresis analysis.

Biopolymer interactions have many potential applications in pharmaceutical, cosmetic, nutraceutical, and functional food industries. Attractive interactions between proteins and polysaccharides can lead to the formation of complexes. Binding parameters of beta-lactoglobulin (beta-lg)/pectin complexes were determined using frontal analysis continuous capillary electrophoresis and the overlapping binding site model. At pH 4, approximately 23 beta-lg molecules were cooperatively complexed on low-methoxyl pectin, where each beta-lg molecule covered an average of 12 galacturonic acid residues. The calculated binding constant was 1431 M(-1). The interactions between pectin and four selected peptides located on the outer surface of the beta-lg were investigated in order to identify which part of the protein was likely to interact with the pectin. The peptide beta-lg 132-148, which corresponds to the alpha-helix zone, and the peptides beta-lg 76-83, 41-60, and 1-14 would be involved in the interaction with the pectin.

Binding Sites↗

Biochemical properties of black drum and sheepshead seabream skin collagen.

Acid-soluble collagen (ASC) and pepsin solubilized collagen (PSC) were isolated from the skins of black drum (Pogonias cromis) and sheepshead seabream (Archosargus probatocephalus) harvested in the Gulf of Mexico coastal waters. The yields of ASCs on dry basis from black drum and sheepshead were estimated at 2.3 and 2.6%, and the yields of PSCs were 15.8 and 29.3%, respectively. Analyses of molecular weight profile, amino acid composition, and secondary structure showed that the skin collagens from both species were typical type-I collagen. The molecular mass of alpha(1) and alpha(2) subunits, as determined by SDS-PAGE using Tris-Acetate gels, was 127 kDa and 116 kDa, respectively. The amino acid composition of ASC and PSC for both species was closer to calf skin ASC than to cod skin ASC. Thermal denaturation temperatures, measured by melting point using circular dichroism, gave the following values: black drum ASC, 34.2 degrees C; sheepshead ASC, 34.0 degrees C; black drum PSC, 35.8 degrees C; sheepshead PSC, 34.3 degrees C. The literature value for the heat stability of calf skin collagen is 36.3 degrees C. The potentials of collagens from black drum and sheepshead skins in the functional food, healthcare, and pharmaceutical industries are discussed.

Amino Acids↗

Black rice (Oryza sativa L. indica) pigmented fraction suppresses both reactive oxygen species and nitric oxide in chemical and biological model systems.

Anthocyanins, present in fruits and vegetables as natural colorants, have been well characterized to possess bioactive properties. Anthocyanin components extracted from black rice (Oryza sativa L. indica) separated by gel filtration and identified using LC-MS were cyanidin 3-glucoside and peonidin 3-glucoside. A standardized extract of black rice pigmented fraction (BRE) containing known proportions of cyanidin 3-glucoside and peonidin 3-glucoside exhibited marked antioxidant activities and free radical scavenging capacities in a battery of in vitro model systems. Significant (p < 0.05) prevention of supercoiled DNA strand scission induced by reactive oxygen species (specifically, peroxyl radical and hydroxyl radicals) and suppression of the oxidative modification of human low-density lipoprotein was obtained with BRE. In addition, BRE reduced (p < 0.05) the formation of nitric oxide by suppressing inducible nitric oxide synthase expression in murine macrophage RAW264.7 cells, without introducing cell toxicity. The results of this study show that black rice contains anthocyanin pigments with notable antioxidant and anti-inflammatory properties for potential use in nutraceutical or functional food formulations.

Anthocyanins↗

In vitro inhibition of the activation of Pro-matrix Metalloproteinase 1 (Pro-MMP-1) and Pro-matrix metalloproteinase 9 (Pro-MMP-9) by rice and soybean Bowman-Birk inhibitors.

The in vitro inhibitory activity of the rice Bowman-Birk inhibitor (rBBI) or soybean Bowman-Birk inhibitor (sBBI) against trypsin-catalyzed activation of pro-matrix metalloproteinase 1 or 9 (pro-MMP-1 or pro-MMP-9), respectively, was investigated using electrophoresis with silver staining, heparin-enhanced zymography, biotinylated gelatin, Biotrak assay, and fluorescence quenched substrate hydrolysis. rBBI at concentrations of 0.08-0.352 mg/mL dose-dependently inhibited the in vitro activation of 45 microg/mL pro-MMP-1 by trypsin. Heparin-enhanced zymography analysis of pro-MMP-1, trypsin-activated MMP-1, and a mixture of pro-MMP-1-trypsin-rBBI showed clear zones associated with trypsin-activated MMP-1 and the absence of clear zones in lanes containing pro-MMP-1 or a mixture of pro-MMP-1, trypsin, and rBBI. The results of the Biotrak assay also indicated that rBBI dose-dependently suppressed the activation of pro-MMP-1 by trypsin. sBBI dose-dependently inhibited the activation of 100 microg/mL of pro-MMP-9 by trypsin. Biotinylated gelatin assays demonstrated that pro-MMP-9 or pro-MMP-9 in the presence of trypsin and BBI did not hydrolyze gelatin, whereas p-aminophenylmercury acetate (APMA)-activated MMP-9 and trypsin-activated MMP-9 caused significant hydrolysis of gelatin. Quenched fluorescence substrate hydrolysis for total MMP activity showed that pro-MMP-1 or pro-MMP-9 did not hydrolyze the substrate Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2; active MMP-1 or MMP-9 hydrolyzed the substrate, but lower substrate hydrolysis was obtained when pro-MMP-1 or pro-MMP-9 was incubated with trypsin in the presence of increasing concentrations of rBBI. The results are discussed in light of the role of MMP-1 and MMP-9 in the process of angiogenesis and the potential of rBBI or sBBI as a functional food ingredient.

Biotinylation↗

Improved method to obtain pure alpha-galactosides from lupin seeds.

Improvement of a previously described method of purification of alpha-galactosides, members of the raffinose family of oligosaccharides (RFOs), has been developed for lupins. The considerable amount of sucrose present in the RFO preparations obtained by the previous method has been removed by modifying the purification stage on diatomaceous earth and charcoal. The present method allows for the preparation of high-purity RFOs containing approximately 99.4% of RFOs in the form of a white fine powder, which provides new perspectives for the production of pure alpha-galactoside preparations for their use as prebiotics in functional foods.

Chromatography, High Pressure Liquid↗