Dietary supplementation with soy isoflavones or replacement with soy proteins prevents hepatic lipid droplet accumulation and alters expression of genes involved in lipid metabolism in rats.

Accumulation of hepatic lipid droplet (HLD) is the hallmark pathology of non-alcoholic fatty liver disease (NAFLD). This study examined the effects of soy isoflavones (ISF) and different amounts of soy proteins on the accumulation of HLD, lipid metabolism and related gene expression in rats. Weanling Sprague-Dawley rats were fed diets containing either 20 % casein protein without (D1) or with (D2) supplemental ISF (50 mg/kg diet) or substitution of casein with increasing amounts of alcohol-washed soy protein isolate (SPI, 5, 10, and 20 %; D3, D4, D5) for 90 days. Dietary casein (20 %) induced accumulation of HLD in female, but not in male rats. Both soy proteins and ISF remarkably prevented the formation of HLD. Soy proteins lowered hepatic total cholesterol and triglyceride in a dose-dependent manner. Interestingly, soy proteins but not ISF significantly increased free fatty acids in the liver of the female rats compared to D1. Proteomic analysis showed that at least 3 enzymes involved in lipogenesis were down-regulated and 7 proteins related to fatty acid ß-oxidation or lipolysis were up-regulated by soy protein over D1. Additionally, 9 differentially expressed proteins identified were related to amino acid metabolism, 5 to glycolysis and 2 to cholesterol metabolism. Dietary ISF and SPI markedly reduced hepatic-peroxisome-proliferator-activated receptor γ2 (PPARγ2) and fat-specific protein 27 (FSP27) in female rats. Overall, this study has shown that partial or full replacement of dietary casein by soy protein or supplementation with soy ISF can effectively prevent the accumulation of HLD. The potential molecular mechanism(s) involved might be due to suppression of lipogenesis and stimulation of lipolysis and down-regulation of PPARγ2 and FSP27. This suggests that consumption of soy foods or supplements might be a useful strategy for the prevention or treatment of fatty liver diseases.

Cohort profile: the maternal-infant research on environmental chemicals research platform.

BACKGROUND: The Maternal-Infant Research on Environmental Chemicals (MIREC) Study was established to obtain Canadian biomonitoring data for pregnant women and their infants, and to examine potential adverse health effects of prenatal exposure to priority environmental chemicals on pregnancy and infant health. METHODS: Women were recruited during the first trimester from 10 sites across Canada and were followed through delivery. Questionnaires were administered during pregnancy and post-delivery to collect information on demographics, occupation, life style, medical history, environmental exposures and diet. Information on the pregnancy and the infant was abstracted from medical charts. Maternal blood, urine, hair and breast milk, as well as cord blood and infant meconium, were collected and analysed for an extensive list of environmental biomarkers and nutrients. Additional biospecimens were stored in the study's Biobank. The MIREC Research Platform encompasses the main cohort study, the Biobank and follow-up studies. RESULTS: Of the 8716 women approached at early prenatal clinics, 5108 were eligible and 2001 agreed to participate (39%). MIREC participants tended to smoke less (5.9% vs. 10.5%), be older (mean 32.2 vs. 29.4 years) and have a higher education (62.3% vs. 35.1% with a university degree) than women giving birth in Canada. CONCLUSIONS: The MIREC Study, while smaller in number of participants than several of the international cohort studies, has one of the most comprehensive datasets on prenatal exposure to multiple environmental chemicals. The biomonitoring data and biological specimen bank will make this research platform a significant resource for examining potential adverse health effects of prenatal exposure to environmental chemicals.

Impact of antinutritional factors in food proteins on the digestibility of protein and the bioavailability of amino acids and on protein quality.

Dietary antinutritional factors have been reported to adversely affect the digestibility of protein, bioavailability of amino acids and protein quality of foods. Published data on these negative effects of major dietary antinutritional factors are summarized in this manuscript. Digestibility and the quality of mixed diets in developing countries are considerably lower than of those in developed regions. For example, the digestibility of protein in traditional diets from developing countries such as India, Guatemala and Brazil is considerably lower compared to that of protein in typical North American diets (54-78 versus 88-94 %). Poor digestibility of protein in the diets of developing countries, which are based on less refined cereals and grain legumes as major sources of protein, is due to the presence of less digestible protein fractions, high levels of insoluble fibre, and/or high concentrations of antinutritional factors present endogenously or formed during processing. Examples of naturally occurring antinutritional factors include glucosinolates in mustard and canola protein products, trypsin inhibitors and haemagglutinins in legumes, tannins in legumes and cereals, gossypol in cottonseed protein products, and uricogenic nucleobases in yeast protein products. Heat/alkaline treatments of protein products may yield Maillard reaction compounds, oxidized forms of sulphur amino acids, D-amino acids and lysinoalanine (LAL, an unnatural nephrotoxic amino acid derivative). Among common food and feed protein products, soyabeans are the most concentrated source of trypsin inhibitors. The presence of high levels of dietary trypsin inhibitors from soyabeans, kidney beans or other grain legumes have been reported to cause substantial reductions in protein and amino acid digestibility (up to 50 %) and protein quality (up to 100 %) in rats and/or pigs. Similarly, the presence of high levels of tannins in sorghum and other cereals, fababean and other grain legumes can cause significant reductions (up to 23 %) in protein and amino acid digestibility in rats, poultry, and pigs. Normally encountered levels of phytates in cereals and legumes can reduce protein and amino acid digestibility by up to 10 %. D-amino acids and LAL formed during alkaline/heat treatment of lactalbumin, casein, soya protein or wheat protein are poorly digestible (less than 40 %), and their presence can reduce protein digestibility by up to 28 % in rats and pigs, and can cause a drastic reduction (100 %) in protein quality, as measured by rat growth methods. The adverse effects of antinutritional factors on protein digestibility and protein quality have been reported to be more pronounced in elderly rats (20-months old) compared to young (5-weeks old) rats, suggesting the use of old rats as a model for assessing the protein digestibility of products intended for the elderly.

High-dose supplemental selenite to male Syrian hamsters fed hypercholesterolaemic diets alters Ldlr, Abcg8 and Npc1l1 mRNA expression and lowers plasma cholesterol concentrations.

The aim of the present study was to elucidate possible cholesterol-lowering mechanism(s) of high-dose supplemental Se in the form of selenite, a known hypocholesterolaemic agent. Male Syrian hamsters (four groups, ten per group) were fed semi-purified diets for 4 weeks containing 0.1 % cholesterol and 15 % saturated fat with selenite corresponding to varying levels of Se: (1) Se 0.15 parts per million (ppm), control diet; (2) Se 0.85 ppm; (3) Se 1.7 ppm; (4) Se 3.4 ppm. Lipids were measured in the bile, faeces, liver and plasma. The mRNA expression of several known regulators of cholesterol homeostasis (ATP-binding cassette transporters g5 (Abcg5) and g8 (Abcg8), 7-hydroxylase, 3-hydroxy-3-methylglutaryl-coenzyme A reductase, LDL receptor (LdLr) and Nieman-Pick C1-like 1 protein (Npc1l1)) were measured in the liver and/or jejunum. Oxysterols including 24-(S)-hydroxycholesterol, 25-hydroxycholesterol and 27-hydroxycholesterol (27-OHC) were measured in the liver. Significantly lower total plasma cholesterol concentrations were observed in hamsters consuming the low (0.85 ppm) and high (3.4 ppm) Se doses. The two highest doses of Se resulted in decreased plasma LDL-cholesterol concentrations and increased mRNA levels of hepatic Abcg8, Ldlr and jejunal Ldlr. Higher hepatic 27-OHC and TAG concentrations and lower levels of jejunal Npc1l1 mRNA expression were noted in the 1.7 and 3.4 ppm Se-treated hamsters. Overall, Se-induced tissue changes in mRNA expression including increased hepatic Abcg8 and Ldlr, increased jejunal Ldlr and decreased jejunal Npc1l1, provide further elucidation regarding the hypocholesterolaemic mechanisms of action of Se in the form of selenite.

Canadians continue to consume too much sodium and not enough potassium.

OBJECTIVES: Excessive sodium (Na) intakes and insufficient potassium (K) intakes are known contributors to hypertension. In July 2010, the Health Canada-led multi-stakeholder Sodium Working Group issued recommendations to lower Na intakes of Canadians. Baseline data and ongoing monitoring are needed. METHODS: Na and K content based on recently analyzed food composite samples from the Canadian marketplace were matched with over 35,000 dietary recalls from the Canadian Community Health Survey (CCHS 2.2). The distributions of usual intakes for Na and K were constructed using SIDE software and estimates by age and sex for the 5th, 10th, 25th, mean, median, 75th, 90th and 95th percentiles were determined. RESULTS: Based on recent analyses of Canadian foods, the majority of Canadians exceeded the Tolerable Upper Intake Level (UL) for Na for their age and sex group, including infants, children, adolescents and adults. In sharp contrast, few had Adequate Intakes (AI) of K. CONCLUSION: Canadians of all ages need to decrease Na intakes below the UL. At the same time, increased consumption of dairy products, fruits and vegetables must be promoted to increase K intakes to current recommendations. Both dietary interventions are required to help lower hypertension in the Canadian population. We provide the first report based on direct analysts of Canadian foods, confirming the high Na and low K intakes of the Canadian population. With its annual sampling program of foods commonly consumed in Canada, the Total Diet Study provides an important sentinel system for monitoring these dietary risk factors for hypertension.

Evidence of oxidative stress in relation to feeding type during early life in premature infants.

Morbidity in the premature (PT) infant may reflect difficult adaptation to oxygen. We hypothesized that feeding including formula feeding (F) and feeding mother's milk (HM) with added fortifier would affect redox status. Therefore, 65 PT infants (birth weight: 1146 ± 261 g; GA: 29 ± 2.5 wk; mean ± SD) were followed biweekly, once oral feeds were introduced. Feeding groups: F (>75% total feeds) and HM (>75% total feeds) were further subdivided according to human milk fortifier (HMF) content of 0-19, 20-49, and ≥ 50%. Oxidative stress was quantified by F2-isoprostanes (F2-IsoPs) in urine, protein carbonyls, and oxygen radical absorbance capacity (ORAC) in plasma. F2-IsoPs (ng/mg creatinine): 0-2 wk, 125 ± 63; 3-4 wk, 191 ± 171; 5-6 wk, 172 ± 83; 7-8 wk, 211 ± 149; 9-10 wk, 222 ± 121; and >10 wk, 183 ± 67. Protein carbonyls from highest [2.41 ± 0.75 (n = 9)] and lowest [2.25 ± 0.89 (n = 12) pmol/µg protein] isoprostane groups did not differ. ORAC: baseline, 6778 ± 1093; discharge, 6639 ± 735 [full term 4 and 12 M, 9010 ± 600 mg (n = 12) TE]. Highest isoprostane values occurred in infants with >50% of their mother's milk fortified. Further research on HMF is warranted.

Effect of calcium on iron absorption in women with marginal iron status.

We measured non-haem Fe absorption with and without added Ca in a short-term feeding study, in thirteen women with marginal Fe status, by the use of a double stable isotope technique. Supplementing 500 mg Ca as calcium carbonate significantly (P = 0.0009) reduced Fe absorption from a single meal from 10.2 % (range 2.2-40.6) to 4.8 % (range 0.7-18.9). A significant inverse correlation in the absence ( - 0.67, P = 0.010) and presence ( - 0.58, P = 0.037) of Ca, respectively, was found between Fe absorption and Fe stores measured by serum ferritin (SF). Wide variation in Fe absorption was observed between individuals in the absence and in the presence of Ca, despite pre-selection of participants within a relatively narrow range of iron stores (SF concentrations). Correction of Fe absorption data based on group mean SF was not found to be useful in reducing the inter-individual variability in iron absorption. It appears that selecting a study group with a narrow initial range of Fe stores does not necessarily reduce the inter-individual variability in Fe bioavailability measurements. These results support the hypothesis that body Fe stores, although an important determinant of dietary Fe absorption, are not the main factor that determines Fe absorption under conditions of identical dietary intake in subjects with low Fe stores.

Antioxidant Supplements Improve Profiles of Hepatic Oxysterols and Plasma Lipids in Butter-fed Hamsters.

Hypercholesterolemic diets are associated with oxidative stress that may contribute to hypercholesterolemia by adversely affecting enzymatically-generated oxysterols involved in cholesterol homeostasis. An experiment was conducted to examine whether the cholesterol-lowering effects of the antioxidants selenium and α-tocopherol were related to hepatic oxysterol concentrations. Four groups of male Syrian hamsters (n = 7-8) were fed high cholesterol and saturated fat (0.46% cholesterol, 14.3% fat) hypercholesterolemic semi-purified diets: 1) Control; 2) Control + α-tocopherol (67 IU all-racemic-α-tocopheryl-acetate/kg diet); 3) Control + selenium (3.4 mg selenate/kg diet); and 4) Control + α-tocopherol + selenium. Antioxidant supplementation was associated with lowered plasma cholesterol concentrations, decreased tissue lipid peroxidation and higher hepatic oxysterol concentrations. A second experiment examined the effect of graded selenium doses (0.15, 0.85, 1.7 and 3.4 mg selenate/kg diet) on mRNA expression of the oxysterol-generating enzyme, hepatic 27-hydroxylase (CYP27A1, EC 1.14.13.15), in hamsters (n = 8-9) fed the hypercholesterolemic diets. Supplementation of selenium at 3.4 mg selenate/kg diet was not associated with increased hepatic 27-hydroxylase mRNA. In conclusion, the cholesterol lowering effects of selenium and α-tocopherol were associated with increased hepatic enzymatically generated oxysterol concentrations, which appears to be mediated via improved antioxidant status rather than increased enzymatic production.

Application of the Dietary Reference Intakes in developing a recommendation for pregnancy iron supplements in Canada.

BACKGROUND: For many pregnant Canadian women, usual iron intakes from food appear to be inadequate compared with Dietary Reference Intake requirement estimates. OBJECTIVE: Dietary intake modeling was undertaken to determine an amount of iron supplementation that would confer acceptably low prevalence of apparently inadequate and apparently excessive intakes. DESIGN: The distribution of usual dietary iron intakes was estimated with the use of 24-h recalls from pregnant women aged 19-50 y in the Canadian Community Health Survey, Cycle 2.2. The prevalence of usual intakes below the Estimated Average Requirement for pregnancy (22 mg/d) or above the Tolerable Upper Intake Level (45 mg/d) was estimated. Iterative modeling with incremental iron supplement was performed to determine a suitable supplement amount. Because the sample of pregnant women was small (148 day 1 recalls), estimates of the tails of the distributions had large SDs, and supporting analyses based on intake data from nonpregnant women (4540 day 1 recalls) were made. RESULTS: Daily supplementation shifted the intake distribution curve without changing its shape. Supplementation with 16 mg iron/d was consistent with low (<3%) prevalence of apparently inadequate intakes. This amount of supplementation should not be associated with an increase in apparently excessive intakes by pregnant women in this population. CONCLUSIONS: On the basis of Dietary Reference Intakes, an iron supplement of 16 mg/d throughout pregnancy is justified as both efficacious and safe for healthy women living in Canadian households. This does not preclude the need for therapeutic iron doses for some individuals on the basis of iron status. The method can be applied to other populations if suitable baseline iron intake data are available.

Determination of total iodine in food samples using inductively coupled plasma-mass spectrometry.

A method was developed and validated for the extraction and determination of total iodine (I) in food composite samples, representing different foods available on the Canadian market, by inductively coupled plasma-mass spectrometry (ICP-MS). Prior to analysis, samples were digested in a closed microwave system using a mixture of nitric acid and perchloric acid. The detection limit for iodine determination was 29 nglg and precisions of 10 and 1.3% were obtained for 10 replicate measurements of 100 and 1000 ng/g standards, respectively. The method was validated using Certified Reference Materials and spike recovery measurements in food samples and was applied for the determination of iodine in a variety of food composite samples from the Canadian Total Diet Study. The high sample throughput of ICP-MS makes the method suitable for analysis of large numbers of food samples with varying matrixes, such as for Total Diet Studies.

Regulatory frameworks for copper considering chronic exposures of the population.

Copper is an essential nutrient that is toxic in excess. Copper intakes from a balanced diet appear to meet the needs of most healthy individuals, because overt deficiency and toxicity are rare. Some uncertainty, however, persists because of limitations in currently available biomarkers used to assess copper status and the paucity of data available to establish tolerable upper levels of intake. Current policies and regulations pertaining to food fortification, nutritional supplements, and drinking water appear to be effective in providing for adequate copper intakes in many populations, although high levels of exposure, through overzealous fortification, supplementation, or drinking water exposure, may be possible under some circumstances. Surveillance and monitoring programs to evaluate copper exposures of human populations should continue and should be refined as new biomarkers become available.

Isotope ratio measurements of iron in blood samples by multi-collector ICP-MS to support nutritional investigations in humans.

With the perspective of embarking on a human study using a double iron (Fe) stable isotope tracer protocol to assess iron bioavailability, investigations were conducted on Fe isotope ratios in blood samples using a VG Axiom Multi-collector ICP-MS. The factors affecting the precision and accuracy of Fe isotopic ratios, such as spectral- and matrix-induced interferences and Fe recoveries from sample preparation, have been identified and optimized. Major polyatomic interferences (e.g., Ar-O, Ar-OH, and FeH) were significantly reduced by using an Aridus nebulizer and desolvating system. Isobaric metal (e.g., (54)Cr(+) on (54)Fe(+) and (58)Ni(+) on (58)Fe(+)) interferences and Ca-oxides and hydroxides were quantitatively removed during chemical purification of blood samples and selective isolation of Fe by anion-exchange resin, after mineralization of the blood samples by microwave digestion. Quantitative recoveries of Fe from different steps of sample preparation were verified using whole blood reference material. Fe isotopic compositions of the samples were corrected for instrumental mass bias by the standard-sample bracketing method using the certified reference standard IRMM-014. External precisions on the order of 0.008-0.05 (% RSD), 0.007-0.015 (% RSD), and 0.03-0.09 (% RSD) were obtained for (54)Fe/(56)Fe, (57)Fe/(56)Fe, and (58)Fe/(56)Fe, respectively, in the blood for three replicate measurements. The level of precision obtained in this work enables the detection of low enrichments of Fe in blood, which is highly desired in nutrition tracer studies.

Impact of dietary protein on lipid metabolism in hamsters is source-dependent and associated with changes in hepatic gene expression.

This study tested the hypothesis that protein source is a factor determining the impact of the diet on lipid metabolism in hamsters. Twenty-eight hamsters of similar body weight were assigned for a period of 8 weeks to one of the following four diets (seven per group) containing either 20 % (w/w) casein (CAS), beef protein (BF), wheat gluten (WG) or soya protein (SOY). The fat composition of the diet was the same (15.5 % w/w) in all groups and provided SFA, MUFA and PUFA representative of the average Canadian diet. After an overnight fast, blood and liver were collected for the measurement of serum lipids, fatty acid composition of liver phospholipids and mRNA levels of selected genes involved in lipid metabolism. WG resulted in lower total cholesterol, HDL-cholesterol and non-HDL-cholesterol but, along with SOY, in higher mRNA levels of cholesterol 7 alpha-hydroxylase and LDL receptor. Furthermore, both WG and SOY resulted in lower 18 : 3n-3, 20 : 4n-6, total n-6 PUFA, 18 : 1n-9 and total MUFA, but higher 22 : 6n-3, total n-3 PUFA, 22 : 6n-3/18 : 3n-3 and 22 : 5n-3/18 : 3n-3 ratios in liver phospholipids, and higher hepatic Delta6-desaturase mRNA levels. These results show that the impact of dietary protein on lipid metabolism is source-dependent and associated with changes in mRNA abundances of key hepatic enzymes and receptors.

An overview of methods for assessment of iron bioavailability from foods nutritionally enhanced through biotechnology.

Iron deficiency and iron deficiency anemia continue to be significant public health problems worldwide. While supplementation and fortification have been viable means to improve iron nutriture of the population in developed countries, they may be less successful in developing regions for a number of reasons, including complexities in distribution and consumer compliance. Biofortification of staple crops, through conventional plant breeding strategies or modern methods of biotechnology, provides an alternative approach that may be more sustainable once initial investments have been made. Three types of biofortification strategies are being essayed, singly or in combination: increasing the total iron content of edible portions of the plant, decreasing the levels of inhibitors of iron absorption, and increasing the levels of factors that enhance iron absorption. Bioavailability is a key concept in iron nutrition, particularly for nonheme iron such as is found in these biofortified foods. An overview is presented of methods for evaluation of iron bioavailability from foods nutritionally enhanced through biotechnology.

Sparing effects of selenium and ascorbic acid on vitamin C and E in guinea pig tissues.

BACKGROUND: Selenium (Se), vitamin C and vitamin E function as antioxidants within the body. In this study, we investigated the effects of reduced dietary Se and L-ascorbic acid (AA) on vitamin C and alpha-tocopherol (AT) status in guinea pig tissues. METHODS: Male Hartley guinea pigs were orally dosed with a marginal amount of AA and fed a diet deficient (Se-D/MC), marginal (Se-M/MC) or normal (Se-N/MC) in Se. An additional diet group (Se-N/NC) was fed normal Se and dosed with a normal amount of AA. Guinea pigs were killed after 5 or 12 weeks on the experimental diets at 24 and 48 hours post AA dosing. RESULTS: Liver Se-dependent glutathione peroxidase activity was decreased (P < 0.05) in guinea pigs fed Se or AA restricted diets. Plasma total glutathione concentrations were unaffected (P > 0.05) by reduction in dietary Se or AA. All tissues examined showed a decrease (P < 0.05) in AA content in Se-N/MC compared to Se-N/NC guinea pigs. Kidney, testis, muscle and spleen showed a decreasing trend (P < 0.05) in AA content with decreasing Se in the diet. Dehydroascorbic acid concentrations were decreased (P < 0.05) in several tissues with reduction in dietary Se (heart and spleen) or AA (liver, heart, kidney, muscle and spleen). At week 12, combined dietary restriction of Se and AA decreased AT concentrations in most tissues. In addition, restriction of Se (liver, heart and spleen) and AA (liver, kidney and spleen) separately also reduced AT in tissues. CONCLUSION: Together, these data demonstrate sparing effects of Se and AA on vitamin C and AT in guinea pig tissues.

The iron status of Canadian adolescents and adults: current knowledge and practical implications.

OVERVIEW: Iron is an essential nutrient, playing a central role in oxygen transport and cellular energy metabolism. The importance of ensuring adequate bioavailable dietary iron stems from the severe consequences associated with iron deficiency (ID) and anemia, including reduced immune function and resistance to infection, developmental delays and irreversible cognitive deficits in young children, impaired physical work performance, and adverse pregnancy outcomes. SPECIFIC POPULATIONS: Poor dietary iron intake and ID exist in Canada, particularly in women of reproductive age. Data from the provincial nutrition surveys suggest that the prevalence of inadequate iron intakes (and low intakes of absorbable iron) among women under 50 years of age is over 10%, which may reflect poor iron status. Teenage girls are at risk for low iron stores because of the adolescent growth spurt and the onset of menstruation; those who are vegetarian are at even greater risk. CONCLUSIONS: The Canadian diet has changed so that grain products are now the main source of dietary iron for all age groups. The public must be educated to ensure the consumption of adequate quantities of bioavailable iron and enhancing factors such as vitamin C. Industry, government, and health professionals must work together to promote healthy eating patterns and the selection of appropriate foods.

Limited effects of combined dietary copper deficiency/iron overload on oxidative stress parameters in rat liver and plasma.

Copper (Cu) deficiency decreases the activity of Cu-dependent antioxidant enzymes such as Cu,zinc-superoxide dismutase (Cu,Zn-SOD) and may be associated with increased susceptibility to oxidative stress. Iron (Fe) overload represents a dietary oxidative stress relevant to overuse of Fe-containing supplements and to hereditary hemochromatosis. In a study to investigate oxidative stress interactions of dietary Cu deficiency with Fe overload, weanling male Long-Evans rats were fed one of four sucrose-based modified AIN-93G diets formulated to differ in Cu (adequate 6 mg/kg diet vs. deficient 0.5 mg/kg) and Fe (adequate 35 mg/kg vs. overloaded 1500 mg/kg) in a 2 x 2 factorial design for 4 weeks prior to necropsy. Care was taken to minimize oxidation of the diets prior to feeding to the rats. Liver and plasma Cu content and liver Cu,Zn-SOD activity declined with Cu deficiency and liver Fe increased with Fe overload, confirming the experimental dietary model. Liver thiobarbituric acid reactive substances were significantly elevated with Fe overload (pooled across Cu treatments, 0.80+/-0.14 vs. 0.54+/-0.08 nmol/mg protein; P<.0001) and not affected by Cu deficiency. Liver cytosolic protein carbonyl content and the concentrations of several oxidized cholesterol species in liver tissue did not change with these dietary treatments. Plasma protein carbonyl content decreased in Cu-deficient rats and was not influenced by dietary Fe overload. The various substrates (lipid, protein and cholesterol) appeared to differ in their susceptibility to the in vivo oxidative stress induced by dietary Fe overload, but these differences were not exacerbated by Cu deficiency.

Effects of antinutritional factors on protein digestibility and amino acid availability in foods.

Digestibility of protein in traditional diets from developing countries such as India, Guatemala, and Brazil is considerably lower compared to that of protein in typical North American diets (54-78 versus 88-94%). The presence of less digestible protein fractions, high levels of insoluble fiber, and high concentrations of antinutritional factors in the diets of developing countries, which are based on less refined cereals and grain legumes as major sources of protein, are responsible for poor digestibility of protein. The effects of the presence of some of the important antinutritional factors on protein and amino digestibilities of food and feed products are reviewed in this chapter. Food and feed products may contain a number of antinutritional factors that may adversely affect protein digestibility and amino acid availability. Antinutritional factors may occur naturally, such as glucosinolates in mustard and rapeseed protein products, trypsin inhibitors and hemagglutinins in legumes, tannins in legumes and cereals, phytates in cereals and oilseeds, and gossypol in cottonseed protein products. Antinutritional factors may also be formed during heat/alkaline processing of protein products, yielding Maillard compounds, oxidized forms of sulfur amino acids, D-amino acids, and lysinoalanine (LAL, an unnatural amino acid derivative). The presence of high levels of dietary trypsin inhibitors from soybeans, kidney beans, or other grain legumes can cause substantial reductions in protein and amino acid digestibilities (up to 50%) in rats and pigs. Similarly, the presence of high levels of tannins in cereals, such as sorghum, and grain legumes, such as fababean (Vicia faba L.), can result in significantly reduced protein and amino acid digestibilities (up to 23%) in rats, poultry, and pigs. Studies involving phytase supplementation of production rations for swine or poultry have provided indirect evidence that normally encountered levels of phytates in cereals and legumes can reduce protein and amino acid digestibilities by up to 10%. D-amino acids and LAL formed during alkaline/heat treatment of proteins such as casein, lactalbumin, soy protein isolate, or wheat proteins are poorly digestible (less than 40%), and their presence can reduce protein digestibility by up to 28% in rats and pigs. A comparison of the protein digestibility determination in young (5-week) versus old (20-month) rats suggests greater susceptibility to the adverse effects of antinutritional factors in old rats than in young rats. Therefore, the inclusion of protein digestibility data obtained with young rats, as the recommended animal model, in the calculation of PDCAAS (Protein Digestibility-Corrected Amino Acid Score) may overestimate protein digestibility and quality of products, especially those containing antinutritional factors, for the elderly. For products specifically intended for the elderly, protein digestibility should be determined using more mature rats.