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Short-term and long-term in vivo exposure to an ephedra- and caffeine-containing metabolic nutrition system does not induce cardiotoxicity in B6C3F1 mice.

Although conventional biomedical research has largely focused on mechanisms of weight loss and genetic aspects of obesity, most medical solutions are plagued by side-effects and fraught with complex questions. As a consequence, consumers are seriously considering herbal products, nutraceuticals and functional foods as alternatives to conventional medications. This is evidently driven by a growing consumer understanding of diet/disease links, aging-related consequences, rising health care costs, and advances in food technology and nutrition. This study investigated the effects of up to 12 months exposure to a multinutrient and botanical extract supplement (Metabolic Nutrition System Orange (MNSO) - sold by AdvoCare, Carrollton, TX, USA) at five dietary concentrations on serum biochemistry and target organ histopathology of the hearts of B6C3F1 mice. The MNSO is a unique combination of vitamins, minerals, omega-3 fatty acids and herbal extracts designed to provide a strong foundation of nutritional support, and to enhance thermogenesis and perception of energy. The MNSO contain extracts of citrus, ephedra, guarana, gingko, green tea and Ocimum. In this study, female B6C3F1 mice were fed control (-MNSO) or MNSO (one time to ten times, one time = daily human dose) diets. Animals were sacrificed after 4, 8 and 12 months', at which time blood was collected for serum chemistry analysis, and hearts were prepared for histopathology and tissue biochemistry. Food consumption and body weight changes were also monitored throughout the study. The MNSO exposure did not significantly affect any of the cardiosensitive enzymes [including creatine kinase (CK), lactate dehydrogenase (LDH) and aspartate aminotransferase (AST)] and normal histopathological architecture of the heart was observed. Although animals given the MNSO diet consumed more food, they were relatively leaner and more active compared to controls. The results indicate that ingestion of ephedra and caffeine for one year in the doses used as part of a comprehensive metabolic nutrition system does not significantly alter normal serum chemistry or induce any irreversible histological changes in the mouse heart, since this study employed up to ten times the normal human consumption dose of ephedra and the metabolic nutrition system.

Animals↗

Potential preventive strategies and therapies in urinary tract infection.

There are perhaps five strategies either presently advocated or under investigation for prevention of recurrent urinary tract infection (UTI): antibiotics, including natural peptides; functional foods; vaccines; probiotics; and miscellaneous, including avoidance of spermicides and maintenance of good hygiene. It is not possible to state the proportion of patients using antibiotics versus foods such as cranberry or using alternative approaches such as avoidance of spermicides. The majority of women who are referred to specialists will be prescribed long-term, low-dose antibiotics. However, given the magnitude of the problem, it is safe to state that large numbers of women are at least experimenting with alternative remedies such as drinking of cranberry juice or ingestion of herbal remedies with a view to enhancing their immune response. Vaccine development remains a long way from human use and has yet to be developed for organisms other than Escherichia coli. The use of probiotics of restore the normal vaginal flora and provide a competitive bacterial barrier to pathogens is close to becoming available as an alternative preventive approach. The next decade should see the introduction of new methods for reduction of the high incidence of UTI and better management of recurring urogenital infections.

Anti-Bacterial Agents↗

Attention, psychomotor functions and age.

Nutrition might play an important role to ameliorate or to buffer age-related declines in attention and psychomotor functions. The assessment of nutritional effects in aged subjects has to take into account that attention and psychomotor functions can be subdivided in different functions that are differentially affected by age. This paper gives an overview of changes in different facets of attention and psychomotor functions beyond fifty as well as assessment methods for attention and psychomotor performance. It also provides a review of models to explain the pattern of changes with increasing age, and discusses the problems of high performance variance and of age related confounding variables like health status. Two different approaches are discussed that analyse a performance profile and an experimentally oriented functional microanalysis of changes in performance with respect to the effects of nutrition on attention and psychomotor functions. Addressed are examples of missing age-related deficits or even age-related superiority. Caffeine and Ginseng are considered as examples to enhance performance in older persons. Results are in accordance with data on the positive role of physical fitness for mental performance in older persons. Performance of older persons can well be enhanced by functional food components or nutritional supplementation. The effects are comparable to the effects obtained in younger groups, while there is only weak evidence for specific compensatory effects in aged persons. Finally the role of nutrition for the processes of healthy aging is discussed.

Aged↗

Production of human caseinomacropeptide in recombinant Saccharomyces cerevisiae and Pichia pastoris.

Caseinomacropeptide is a polypeptide of 64 amino acid residues (106-169) derived from the C-terminal part of the mammalian milk k-casein. This macropeptide has various biological activities and is used as a functional food ingredient as well as a pharmaceutical compound. The gene encoding the human caseinomacropeptide (hCMP) was synthesized and expressed with an alpha-factor secretion signal in the two yeast strains, Saccharomyces cerevisiae and Pichia pastoris. The complete polypeptide of the recombinant hCMP was produced and secreted in a culture medium by both the strains, but the highest production was observed in S. cerevisiae with a galactose-inducible promoter. In a fed-batch bioreactor culture, 2.5 g/l of the recombinant hCMP was obtained from the S. cerevisiae at 97 h.

Biomass↗

Temperature-enhanced alumina HPLC method for the analysis of wax esters, sterol esters, and methyl esters.

Previous attempts at separating nonpolar lipid esters (including wax esters, sterol esters, and methyl esters) have achieved only limited success. Among the several normal-phase methods tested, a single recent report of a method employing an alumina column at 30 degrees C with a binary gradient system was the most promising. In the current study, modification of the alumina method by increasing the column temperature to 75 degrees C improved the separation of standards of wax esters and sterol esters. Elevated column temperature also enhanced the separation of FAME with differing degrees of unsaturation. Evidence was also presented to indicate that the method similarly separated phytosterol esters, based on their levels of unsaturation. With the increased interest in phytosterol- and phytostanol-ester enriched functional foods, this method should provide a technique to characterize and compare these products.

Aluminum Oxide↗

Phospholipase D-catalyzed synthesis of novel phospholipid-phytosterol conjugates.

In the present study, plant sterols were modified to form amphiphiles by synthesizing phospholipid derivatives from them so they could be formulated in different functional foods and possibly improve their effects as therapeutic agents. The new phosphatidyl derivatives, phosphatidyl-sitosterols, were synthesized by the transfer reaction of a phosphatidyl residue from PC to beta-sitosterol by phospholipase D (PLD EC 3.1.4.4) from Streptomyces sp. in a biphasic medium.

Catalysis↗

Maternal dietary Alpine butter intake affects human milk: fatty acids and conjugated linoleic acid isomers.

Consumption of CLA by lactating women affects the composition of their milk, but the pattern of the different CLA isomers is still unknown. We determined the effects of short maternal supplementation with CLA-rich Alpine butter on the occurrence of FA and CLA isomers in human milk. In an open randomized controlled study with a two-period cross-over design, milk FA and CLA isomer concentrations were measured on postpartum days > or = 20 in two parallel groups of lactating women before, during, and after consumption of defined quantities of Alpine butter or margarine with comparable fat content (10 d of butter followed by 10 d of margarine for one group, and vice versa in the other). In the 16 women who completed the study (8/group), Alpine butter supplementation increased the C16 and C18 FA, the sum of saturated FA, the 18:1 trans FA, and the trans FA with CLA. The CLA isomer 18:2 c9,t11 increased by 49.7%. Significant increases were also found for the isomers t9,t11, t7,c9, t11,c13, and t8,c10 18:2. The remaining nine of the total 14 detectable isomers showed no changes, and concentrations were <5 mg/100 g fat. A breastfeeding mother can therefore modulate the FA/CLA supply of her child by consuming Alpine butter. Further studies will show whether human milk containing this FA and CLA isomer pattern acts as a functional food for newborns.

Adult↗

Nutritional interventions in cardiovascular disease: new challenges and opportunities.

New opportunities for control of cardiovascular disease (CVD) through nutritional interventions can be divided into three areas: nutritional intervention on newly identified causes of CVD; nutritional strategies to more efficiently or equitably employ current, proven interventions; and nutritional approaches to reduce costs and improve effectiveness so that benefits can be expanded to more of the population. Examples of newly identified causes of CVD amenable to nutritional intervention include fetal/early childhood malnutrition and gene-environment interactions. Examples of more efficient or equitable nutritional strategies include better nutrition education for healthcare providers, patient education resources on the Internet, and widely available functional foods or nutriceuticals. Nutritional approaches to improve cost-effectiveness include better availability of foods associated with reduced CVD risk (eg, fruits, vegetables, fish) and supplementation of the American diet. The advances in nutritional strategies described in this monograph have the potential for population-wide CVD risk reduction.

Cardiovascular Diseases↗

Cholesterol-lowering action of plant sterols.

Plant sterols have an extended history of use as cholesterol-lowering agents. Until the 1970s, the principal interest in plant sterols lay in effects of sitosterol, but over the past decade interest has reemerged in using plant sterols in functional foods. Hydrogenated plant sterols have been shown efficacious in lowering lipid levels, inhibiting cholesterol absorption and regressing plaque in animals. Hydrogenated versus unhydrogenated plant sterol esters have been demonstrated to possess equal efficacy in low-density lipoprotein cholesterol lowering in humans. Unhydrogenated plants sterol esters show consistency in cholesterol-lowering across dosage levels in humans. Unhydrogenated, unesterifed plant sterols yield similar low-density lipoprotein cholesterol lowering efficacy as achieved with hydrogenated sitostanol esters. Solubility and miscibility are likely more important determinants in cholesterol-lowering potential of plant sterols than their specific composition.

Adsorption↗

Nutrition therapy for dyslipidemia.

National guidelines indicate patients with elevated low- density lipoprotein cholesterol should consume less than 7% of calories from saturated fat and less than 200 mg of cholesterol. Trans fatty acids should also be limited. Incorporation of functional foods, such as stanol-containing margarine, soy products, and soluble fiber-rich cereals and vegetables can provide further benefit. In addition to weight loss and physical activity, individuals with hypertriglyceridemia benefit from a diet moderate in fat and carbohydrate rather than a low-fat diet. Including monounsaturated or omega-3 fatty acids lowers serum triglycerides. Many of the dietary strategies to optimize serum lipids also contribute to glycemic control in patients with diabetes mellitus.

Cholesterol, Dietary↗

The lunar environment as a fractional-gravity biological laboratory.

A quarter of a century ago men stepped upon the lunar surface and established the possibility of human expansion beyond Earth. When humans return to the moon to occupy it with greater permanency, an applied lunar biological laboratory would provide a means of conducting experiments on the long-term effects of fractional gravity in animals and plants and provide necessary data to enhance the health, safety and well-being of lunar workers and inhabitants. In-depth studies can go beyond zero-g observations, on-orbit centrifuge studies, and ground-based research providing important insight into continuous 1/6-g effects on biological systems. Studies concentrating on development, gravity sensing, and adaptation/readaptation would provide preliminary data on whether long-term fractional gravity is detrimental or compromising to fundamental biological function. Food production research in 1/6-g would provide important information for on site application to improve the yield and quality of food (animal and plant) produced in the unique lunar environment. The purpose of this paper is to discuss some examples of the major gravitational biology areas that could be studied on the moon and applied to lunar population needs utilizing lunar biological facilities and continuous fractional gravity.

Adaptation, Physiological↗

Metabolic disposition in rats of regular and enzymatically depolymerized sodium carboxymethylcellulose.

Partially enzyme-hydrolysed sodium carboxymethylcellulose (CMC-ENZ), which holds promise as a new, functional food ingredient, is obtained from sodium carboxymethylcellulose (CMC) by enzymatic hydrolysis with a cellulase preparation from Trichoderma longibrachiatum. In the safety evaluation of CMC-ENZ, a comparative disposition study on 14C-labelled CMC and CMC-ENZ was conducted in conventionally kept rats. The 14C label was in the two C atoms of the carboxymethyl group. Two groups of four male and four female rats each were fed diets with 5% unlabelled CMC or CMC-ENZ for a 2-wk adaptation period. A single oral dose of 14C-CMC or 14C-CMC-ENZ solution was then given by gavage (500 mg/kg body weight). Respiratory CO2, urine and faeces were collected at regular intervals, and after 120 hr organs, tissues and the carcass were sampled as well. For both experimental groups, total mean recovery of 14C was 98%, about 95% of the label being excreted with the faeces, 2% or less in the urine, 1% or less with CO2 and a small fraction being retained in the body (CMC, 0.58%; CMC-ENZ, 0.75%). Tissue retention of 14C was highest in the liver of rats of both experimental groups. Only about 49 and 65% of the faecal 14C was extracted with water in the 14C-CMC and 14C-CMC-ENZ dosed rats, respectively. Gel permeation chromatography (GPC) of the dosing solutions and the faecal extracts revealed that CMC is depolymerized during intestinal passage whereas CMC-ENZ is excreted nearly unchanged. Consequently, the molecular weight distribution of the 14C-CMC and 14C-CMC-ENZ faecal excretion products was similar. It is concluded that there is no toxicologically relevant difference between the disposition of CMC and CMC-ENZ.

Administration, Oral↗

Subchronic oral toxicity study with regular and enzymatically depolymerized sodium carboxymethylcellulose in rats.

Enzymatically depolymerized sodium carboxymethylcellulose (CMC-ENZ) is a new functional food ingredient which has a lower molecular weight and viscosity than regular sodium carboxymethylcellulose (CMC). Both compounds are known not to be absorbed to a significant extent, and the human safety of CMC as a thickening agent and stabilizer in food is well established. In the present study, the subchronic oral toxicity of CMC-ENZ was examined and compared with that of CMC in Wistar rats. Seven groups of 20 rats/sex were fed diets with 0 (controls), 2.5, 5 and 10% CMC and 2.5, 5 and 10% CMC-ENZ for a 3-month period. There was only one death that was unrelated to the treatment. Water intake, urine production and urinary sodium excretion increased with increasing doses of CMC and CMC-ENZ due to their sodium content of about 7-8%. The treatment-related occurrence of diarrhoea and caecal enlargement in the mid- and high-dose groups, a slight increase of plasma alkaline phosphatase, and increased urinary calcium and citrate excretions were considered to be generic effects that typically are observed in rodent studies with low digestible carbohydrates. The increased occurrence of nephrocalcinosis and hyperplasia of the urothelial epithelium in some of the treated groups was interpreted as an indirect consequence of a more alkaline urine coupled with an increased calcium excretion. As the frequency and severity of all these changes did not differ between corresponding CMC and CMC-ENZ dose groups, it is concluded that the two products have a similar toxicological profile.

Administration, Oral↗

Moderately elevated plant sterol levels are associated with reduced cardiovascular risk--the LASA study.

Functional foods with supplementation of plant sterols are already used by millions of people. However, at the same time it is current scientific thinking that elevation of plant sterols in the circulation causes coronary heart disease. Therefore, this study aimed to define the risk for coronary heart disease associated with moderately high plant sterol plasma levels in a cohort of elderly. In this study, we evaluated the association between plant sterols and coronary heart disease in a cohort of 1242 subjects older than 65 years, participating at the Longitudinal Aging Study Amsterdam (LASA). Concentrations of sitosterol, campesterol, brassicasterol and stigmasterol were assessed using highly sensitive and specific gas chromatography-mass spectrometry-selected ion-monitoring. Plant sterol concentrations (and their ratios to cholesterol) were slightly, however, significantly lower in patients with coronary heart disease. Moreover, high plasma concentrations of a marker plant sterol, sitosterol, were associated with a markedly reduced risk for coronary heart disease (OR 0.78, CI 0.62-0.98, p<0.05). In contrast neither plant stanols (sitostanol or campestanol) nor the cholesterol synthesis markers (lathosterol, lanosterol and desmosterol) nor their ratios to cholesterol were significantly different in the study groups. These data suggest that plant sterols could have neutral or even protective effects on development of coronary heart disease, which have to be confirmed in interventional trials.

Aged↗

Inhibition of the expression and activity of cyclooxygenase-2 by chicory extract.

Chicory is a major source of fructans with reported prebiotic-bifidogenic properties. In the present study, the potential anti-inflammatory activities of chicory were investigated. Ethyl acetate chicory root extract produced a marked inhibition of prostaglandin E(2) (PGE(2)) production in human colon carcinoma HT29 cells treated with the pro-inflammatory agent TNF-alpha. Two independent mechanisms of action were identified: (1) a drastic inhibition of the induction by TNF-alpha of cyclooxygenase 2 (COX-2) protein expression and (2) a direct inhibition of COX enzyme activities with a significantly higher selectivity for COX-2 activity. The inhibition of TNF-alpha-dependent induction of COX-2 expression was mediated by an inhibition of NF-kappaB activation. A major sesquiterpene lactone of chicory root, the guaianolide 8-deoxylactucin, was identified as the key inhibitor of COX-2 protein expression present in chicory extract. Altogether, the data presented strongly support chicory root as a promising source of functional food ingredient, combining prebiotic and anti-inflammatory properties.

Anti-Inflammatory Agents↗

A novel regulatory pathway for cholesterol degradation via lactostatin.

Our group previously discovered a novel hypocholesterolemic pentapeptide (IIAEK: Ile-Ile-Ala-Glu-Lys, or what we describe as "lactostatin") derived from bovine milk beta-lactoglobulin. To clarify the mechanism of the hypocholesterolemic action of lactostatin, we screened the target gene and signal transducing pathway induced by lactostatin in HepG2, a human liver cell line. Unexpectedly, we found that water-soluble lactostatin can activate cholesterol 7alpha-hydroxylase (CYP7A1) gene expression. Treatment with mitogen-activated protein kinase (MAPK) inhibitor or calcium (Ca2+) channel blocker blocked this activation. We also found that lactostatin regulates the phosphorylation of extracellular signal-regulated kinase (ERK) and intracellular Ca2+ concentration. Here, we show the involvement of a new regulatory pathway in the calcium-channel-related MAPK signaling pathway of lactostatin-mediated cholesterol degradation. Oligopeptide shows promise as a new molecule for the development of medicines and functional foods to prevent and improve hypercholesterolemia and atherosclerosis.

Calcium Channels↗

Structural requirements for the flavonoid-mediated modulation of glutathione S-transferase P1-1 and GS-X pump activity in MCF7 breast cancer cells.

The objective of this study was to investigate the structural requirements necessary for inhibition of glutathione S-transferase P1-1 (GSTP1-1) and GS-X pump (MRP1 and MRP2) activity by structurally related flavonoids, in GSTP1-1 transfected MCF7 cells (pMTG5). The results reveal that GSTP1-1 activity in MCF7 pMTG5 cells can be inhibited by some flavonoids. Especially galangin was able to inhibit almost all cellular GSTP1-1 activity upon exposure of the cells to a concentration of 25microM. Other flavonoids like kaempferol, eriodictyol and quercetin showed a moderate GSTP1-1 inhibitory potential. For GSTP1-1 inhibition, no specific structural requirements necessary for potent inhibition could be defined. Most flavonoids appeared to be potent GS-X transport inhibitors with IC(50) values ranging between 0.8 and 8microM. Luteolin and quercetin were the strongest inhibitors with IC(50) values of 0.8 and 1.3microM, respectively. Flavonoids without a C2-C3 double bond like eriodictyol, taxifolin and catechin did not inhibit GS-X pump activity. The results of this study demonstrate that the structural features necessary for high potency GS-X pump inhibition by flavonoids are (1) the presence of hydroxyl groups, especially two of them generating the 3',4'-catechol moiety; and (2) a planar molecule due to the presence of a C2-C3 double bond. Other factors, like lipophilicity and the total number of hydroxyl groups do not seem to be dominating the flavonoid-mediated GS-X pump inhibition. To identify the GS-X pump responsible for the DNP-SG efflux in MCF7 cells, the effects of three characteristic flavonoids quercetin, flavone and taxifolin on MRP1 and MRP2 activity were studied using transfected MDCKII cells. All three flavonoids as well as the typical MRP inhibitor (MK571) affected MRP1-mediated transport activity in a similar way as observed in the MCF7 cells. In addition, the most potent GS-X pump inhibitor in the MCF7 cells, quercetin, did not affect MRP2-mediated transport activity. These observations clearly indicate that the GS-X pump activity in the MCF7 cells is likely to be the result of flavonoid-mediated inhibition of MRP1 and not MRP2. Altogether, the present study reveals that a major site for flavonoid interaction with GSH-dependent toxicokinetics is the GS-X pump MRP1 rather than the conjugating GSTP1-1 activity itself. Of the flavonoids shown to be most active especially quercetin is frequently marketed in functional food supplements. Given the physiological levels expected to be reached upon supplement intake, the IC(50) values of the present study point at possible flavonoid-drug and/or flavonoid-xenobiotic interactions especially regarding transport processes involved in toxicokinetics.

Biological Transport↗

Concentration-dependent suppressive effect of shrimp head protein hydrolysate on dehydration-induced denaturation of lizardfish myofibrils.

To utilize fishery waste products as functional food material, the shrimp head protein hydrolysate (SHPH) was produced from three species of shrimp wastes, Northern pink shrimp, Endeavour shrimp and black tiger shrimp, by enzymatic hydrolysis. The SHPH was used as a natural food preservative by adding to lizardfish myofibrils at concentrations ranging from 2.5% to 10%. Their effects on the state of water and the denaturation of myofibrils during dehydration were evaluated. The amount of monolayer and multilayer water in myofibrils containing SHPH were higher than those without SHPH (control). DSC analyses revealed that the amount of unfrozen water increased significantly after addition of SHPH. The Ca-ATPase inactivation rate of myofibrils containing SHPH decreased during dehydration while 5-7.5% concentrations of SHPH exhibited optimum effect regardless of the species. The results implicated that SHPH can be used as an alternative food preservative for suppressive the dehydration-induced denaturation of myofibrils.

Animals↗