Journal of Nutrition current issue

Charles H. Hill (1921-2009) [Biographical Article]

Starcher, B., Rucker, R. B. — Fri, 20 Nov 2009 10:01:58 PST

Genistein and a Soy Extract Differentially Affect Three-Dimensional Bone Parameters and Bone-Specific Gene Expression in Ovariectomized Mice [Biochemical, Molecular, and Genetic Mechanisms]

Zhang, Y., Li, Q., Wan, H.-Y., Helferich, W. G., Wong, M.-S. — Fri, 20 Nov 2009 10:01:58 PST

Soy isoflavone preparations, such as purified genistein and a soy extract (Novasoy), were reported previously to exert beneficial effects on bones. Our purpose in this study was to compare the effects of genistein and Novasoy on 3-dimensional trabecular bone parameters and the expression of bone-specific genes in ovariectomized (OVX) mice. The sham-operated mice were fed the control diet and OVX mice were fed diets containing genistein or Novasoy or the control diet, with or without 17β-estradiol treatment, for 5 wk. Trabecular bone parameters of tibias were measured by microcomputed tomography and gene expression was assayed by real-time PCR. Consumption of diets containing genistein or Novasoy partially prevented the ovariectomy-induced increase in body weight but did not alter the uterus weight of the OVX mice. Novasoy, but not purified genistein, significantly preserved trabecular bone mass, bone volume, and trabecular bone separation in the proximal tibial metaphysis. Purified genistein decreased mRNA expression of receptor activator of nuclear factor-B ligand (RANKL), carbonic anhydrase II, and cathepsin K and enhanced the ratio of osteoprotegrin:RANKL mRNA expression in the tibial head of the OVX mice. In contrast, the diet containing Novasoy suppressed the OVX-induced increase in serum alkaline phosphatase but did not alter bone-specific gene expression of tibia. Our study demonstrated that a soy extract containing a similar level of genistein in the form of Novasoy is more effective than purified genistein in improving tibial trabecular bone quality in OVX mice, but the mechanism of action might be distinct from that of genistein.

Dietary Soy Protein Reduces Cardiac Lipid Accumulation and the Ceramide Concentration in High-Fat Diet-Fed Rats and ob/ob Mice [Biochemical, Molecular, and Genetic Mechanisms]

Fri, 20 Nov 2009 10:01:58 PST

Obesity is an epidemic condition strongly associated with cardiovascular morbidity and mortality. Heart disease secondary to obesity is associated with myocardial steatosis, leading to ceramide synthesis and cell dysfunction in a process known as lipotoxicity. Soy protein has been demonstrated to reduce lipotoxicity in the liver and pancreas in different rodent models of obesity. Thus, our purpose in the present work was to assess the effect of dietary soy protein on cardiac lipid accumulation and ceramide formation during obesity and to evaluate its effect in the following 2 rodent models of obesity: 1) a diet-induced obesity model in Sprague-Dawley rats was produced by feeding rats a control or a high-fat casein or soy protein diet for 180 d; and 2) wild-type and ob/ob mice were fed a casein or soy protein diet for 90 d. Soy protein intake led to lower cholesterol and triglyceride concentrations in the hearts of rats and ob/ob mice in association with a greater PPAR mRNA concentration and a lower level of sterol regulatory element binding protein-1 mRNA than those fed casein. The ceramide concentration was also lower in hearts of rats and ob/ob mice that were fed soy protein in association with lower serine palmitoyl transferase (SPT)-1 and tumor necrosis factor- mRNA concentrations. These results indicate that dietary soy protein can reduce the heart ceramide concentration by reducing the expression of SPT-1, a key enzyme in the formation of this sphingolipid in the heart of obese rodents, and by reducing lipid accumulation. Thus, soy protein consumption may be considered as a dietary therapeutic approach for lipotoxic cardiomyopathy prevention.

Conjugated Linoleic Acid Activates AMP-Activated Protein Kinase and Reduces Adiposity More Effectively When Used with Metformin in Mice [Biochemical, Molecular, and Genetic Mechanisms]

Jiang, S., Wang, Z., Riethoven, J.-J., Xia, Y., Miner, J., Fromm, M. — Fri, 20 Nov 2009 10:01:58 PST

Trans-10, cis-12 (t10c12) conjugated linoleic acid (CLA) reduces lipid levels in adipocytes, but the mechanisms involved are still emerging. The hypotheses of this study were that t10c12 CLA treatment activated AMP-activated protein kinase (AMPK) and that the effectiveness of a low dose of t10c12 CLA would be increased when combined with an AMPK activator. We demonstrated t10c12 CLA, directly or indirectly, activated AMPK and increased the amount of phosphorylated acetyl-CoA carboxylase (ACC) in 3T3-L1 adipocytes. Compound C, a potent inhibitor of AMPK, attenuated the phosphorylation of ACC, integrated stress response (ISR), inflammatory response, reduction in key lipogenic transcription factors, and triglyceride (TG) reduction that otherwise occurred in t10c12 CLA-treated adipocytes. Treatment of adipocytes or mice with a low dose of t10c12 CLA in conjunction with the AMPK activator metformin resulted in more TG loss than treatment with the individual chemicals. Additionally, although an inflammatory response was required for robust TG reduction, the combination of t10c12 CLA with AMPK activators had a similar TG loss with a reduced inflammatory response. A microarray analysis of the transcriptional response to either t10c12 CLA, metformin, or the combination, indicated the responses were very similar, with a correlation coefficient of 0.91 or better for genes in the ISR or lipid-related pathways. Altogether, these results support our hypotheses that t10c12 CLA activates AMPK, directly or indirectly, and that metformin increases the effectiveness of t10c12 CLA in reducing TG amounts in adipocytes.

Diallyl Trisulfide Protects Rats from Carbon Tetrachloride-Induced Liver Injury [Biochemical, Molecular, and Genetic Mechanisms]

Hosono-Fukao, T., Hosono, T., Seki, T., Ariga, T. — Fri, 20 Nov 2009 10:01:59 PST

Alk(en)yl sulfides have been found to be responsible for the anticancer, antithrombotic, and antioxidant effects of garlic. We sought to identify the most potent structure of sulfides that exhibits a hepatoprotective effect against carbon tetrachloride (CCl₄)-induced acute liver injury in rats. Rats were pretreated with diallyl trisulfide (DATS) i.g. at a dose of 500 µmol/kg body weight for 5 d. On d 6, CCl₄ was administered i.g. at a dose of 2.5 mL/kg body weight. Twenty-four hours after CCl₄ administration, rats were killed and plasma and liver samples collected. DATS pretreatment significantly suppressed the CCl₄-induced elevation of plasma aspartate aminotransferase and alanine aminotransferase activities (P < 0.05). Histological observations supported the hepatoprotective effects. Western blot and spectrophotometric analyses indicated that DATS suppressed cytochrome P450 2E1 activity and its protein level and elevated those of glutathione S-transferase. Dipropyl trisulfide (DPTS), which is a saturated alkyl chain analogue of DATS, did not affect CCl₄-induced liver toxicity or drug-metabolizing enzymes. These results suggest that hepatoprotective activity of trisulfides is due to their regulation of drug-metabolizing enzymes. Furthermore, the effects of 6 kinds of alk(en)yl trisulfides, including DATS and DPTS, on phase II enzyme activity were examined in rats. Alk(en)yl trisulfides were administered i.g. (500 µmol/kg body weight) to rats for 5 d. Only the allyl group-containing DATS and allyl methyl trisulfide enhanced these activities.

Reduced Milk Triglycerides in Mice Lacking Phosphoenolpyruvate Carboxykinase in Mammary Gland Adipocytes and White Adipose Tissue Contribute to the Development of Insulin Resistance in Pups [Nutrient Physiology, Metabolism, and Nutrient-Nutrient Interactions]

Fri, 20 Nov 2009 10:01:59 PST

Obesity and type 2 diabetes are growing problems worldwide in adults and children. In this study, we focused on understanding the patterning of insulin resistance as a result of altered perinatal nutrition. We analyzed mice in which the binding site for PPAR was deleted from the promoter of the cytosolic phosphoenolpyruvate carboxykinase gene (Pck1) (PPARE^–/–). We analyzed pups from dams with the same genotype as well as fostered and cross-fostered pups. Pck1 expression and triglyceride concentration in the milk were measured. The PPARE mutation reduced Pck1 expression in white adipose tissue (WAT) to 2.2% of wild type (WT) and reduced Pck1 expression in whole mammary gland tissue to 1% of WT. The female PPARE^–/– mice had reduced lipid storage in mammary gland adipocytes and in WAT, resulting in a 40% reduction of milk triglycerides during lactation. Pups from PPARE^–/– dams had insulin resistance as early as 14 d after birth, a condition that persisted into adulthood. WT pups fostered by PPARE^–/– dams had lower body weights and plasma insulin concentrations compared with WT pups reared by WT dams. PPARE^–/– pups fostered by WT dams had improved glucose clearance compared with pups raised by PPARE^–/– dams. PPARE^+/– and PPARE^–/– dams also patterned newborn pups for reduced growth and insulin resistance in utero. Thus, the in utero environment and altered nutrition during the perinatal period cause epigenetic changes that persist into adulthood and contribute to the development of insulin resistance.

Cardiovascular Disease Risk Biomarkers and Liver and Kidney Function Are Not Altered in Postmenopausal Women after Ingesting an Elderberry Extract Rich in Anthocyanins for 12 Weeks [Nutrient Physiology, Metabolism, and Nutrient-Nutrient Interactions]

Fri, 20 Nov 2009 10:01:59 PST

Growing evidence supports a cardio-protective role for anthocyanins; however, there is limited evidence on their efficacy and safety following the consumption of relatively high but dietarily achievable doses in humans. We conducted a parallel-designed, randomized, placebo-controlled study to examine the effect of chronic consumption of anthocyanins on biomarkers of cardiovascular disease (CVD) risk and liver and kidney function in 52 healthy postmenopausal women (n = 26 in treatment and placebo groups). Volunteers (BMI, 24.7 ± 3.6 kg/m²; age, 58.2 ± 5.6 y) consumed 500 mg/d anthocyanins as cyanidin glycosides (from elderberry) or placebo for 12 wk (2 capsules twice/d). At the beginning (wk 0) and end of the 12-wk intervention, levels of anthocyanins and biomarkers of CVD (inflammatory biomarkers, platelet reactivity, lipids, and glucose) and liver and kidney function (total bilirubin, albumin, urea, creatinine, alkaline phosphatase, alanine aminotransferase, and -glutyl transferase) were assessed in fasted blood. Anthropometric, blood pressure, and pulse measurements were also taken. In addition, postprandial plasma anthocyanins were measured (t = 1, 2, 3 h) following a 500-mg oral bolus dose. After 12 wk of chronic exposure to anthocyanins, there was no significant change in biomarkers of CVD risk and liver and kidney function remained within clinically acceptable ranges. We observed no plasma accumulation of anthocyanins; however, postprandial metabolism increased (P = 0.02). In conclusion, these data suggest that chronic consumption of 500 mg/d of elderberry extract for 12 wk is apparently safe, but ineffective in altering biomarkers of CVD risk in healthy postmenopausal women.

Dietary Sulfur Amino Acid Supplementation Reduces Small Bowel Thiol/Disulfide Redox State and Stimulates Ileal Mucosal Growth after Massive Small Bowel Resection in Rats [Nutrient Physiology, Metabolism, and Nutrient-Nutrient Interactions]

Fri, 20 Nov 2009 10:01:59 PST

Following massive small bowel resection in animal models, the remnant intestine undergoes a dynamic growth response termed intestinal adaptation. Cell growth and proliferation are intimately linked to cellular and extracellular thiol/disulfide redox states, as determined by glutathione (GSH) and GSH disulfide (GSSG) (the major cellular redox system in tissues), and cysteine (Cys) and its disulfide cystine (CySS) (the major redox system in plasma), respectively. The study was designed to determine whether dietary supplementation with sulfur amino acids (SAA) leads to a greater reduction in thiol/disulfide redox state in plasma and small bowel and colonic mucosa and alters gut mucosal growth in an established rat model of short bowel syndrome (SBS). Adult rats underwent 80% jejunal-ileal resection (RX) or small bowel transection (surgical control) and were pair-fed either isonitrogenous, isocaloric SAA-adequate (control) or SAA-supplemented diets (218% increase vs. control diet). Plasma and gut mucosal samples were obtained after 7 d and analyzed for Cys, CySS, GSH, and GSSG concentrations by HPLC. Redox status (E_h) of the Cys/CySS and GSH/GSSG couples were calculated using the Nernst equation. SAA supplementation led to a greater reduction in E_h GSH/GSSG in jejunal and ileal mucosa of resected rats compared with controls. Resected SAA-supplemented rats showed increased ileal adaptation (increased full-thickness wet weight, DNA, and protein content compared with RX control-fed rats; increased mucosal crypt depth and villus height compared with all other study groups). These data suggest that SAA supplementation has a trophic effect on ileal adaptation after massive small bowel resection in rats. This finding may have translational relevance as a therapeutic strategy in human SBS.

Essential Amino Acids Increase MicroRNA-499, -208b, and -23a and Downregulate Myostatin and Myocyte Enhancer Factor 2C mRNA Expression in Human Skeletal Muscle [Nutrient Physiology, Metabolism, and Nutrient-Nutrient Interactions]

Fri, 20 Nov 2009 10:01:59 PST

Essential amino acids (EAA) stimulate muscle protein synthesis in humans. However, little is known about whether microRNAs (miRNA) and genes associated with muscle growth are expressed differently following EAA ingestion. Our purpose in this experiment was to determine whether miRNA and growth-related mRNA expressed in skeletal muscle are up- or downregulated in humans following the ingestion of EAA. We hypothesized that EAA would alter miRNA expression in skeletal muscle as well as select growth-related genes. Muscle biopsies were obtained from the vastus lateralis of 7 young adult participants (3 male, 4 female) before and 3 h after ingesting 10 g of EAA. Muscle samples were analyzed for muscle miRNA (miR-499, -208b, -23a, -1, -133a, and -206) and muscle-growth related genes [MyoD1, myogenin, myostatin, myocyte enhancer factor C (MEF2C), follistatin-like-1 (FSTL1), histone deacytylase 4, and serum response factor mRNA] before and after EAA ingestion using real-time PCR. Following EAA ingestion, miR-499, -208b, -23a, -1, and pri-miR-206 expression increased (P < 0.05). The muscle-growth genes MyoD1 and FSTL1 mRNA expression increased (P < 0.05), and myostatin and MEF2C mRNA were downregulated following EAA ingestion (P < 0.05). We conclude that miRNA and growth-related genes expressed in skeletal muscle are rapidly altered within hours following EAA ingestion. Further work is needed to determine whether these miRNA are post-transcriptional regulators of growth-related genes following an anabolic stimulus.

Single-Protein Casein and Gelatin Diets Affect Energy Expenditure Similarly but Substrate Balance and Appetite Differently in Adults [Nutrient Physiology, Metabolism, and Nutrient-Nutrient Interactions]

Fri, 20 Nov 2009 10:01:59 PST

Increasing the protein content of a diet results in increased satiety and energy expenditure (EE). It is not clear whether the magnitude of these effects differs between proteins differing in concentrations of indispensable amino acids (IAA). We hypothesized that a protein lacking IAA may stimulate appetite suppression and EE and may limit positive protein balance. Therefore, we compared appetite, EE, and substrate balances between gelatin (incomplete protein) and casein (complete protein) in single-protein diets with either 25 or 10% of energy (En%) from protein. During a 36-h stay in a respiration chamber, 23 healthy men (n = 11) and women (n = 12) (BMI, 22.2 ± 2.3 kg/m²; age, 25 ± 7 y) consumed 4 isoenergetic diets: 25 En% (25/20/55 En% protein/fat/carbohydrate) and 10 En% (10/35/55 En% protein/fat/carbohydrate) casein or gelatin diet in a randomized crossover design. For 3 d before the study, participants consumed a diet at home with similar macronutrient distribution as the diet they would receive during the subsequent stay in the chamber. Hunger was suppressed 44% more (P < 0.05) and protein balance was more negative when consuming the 10 En% gelatin diet (–0.17 ± 0.03 MJ/d) compared with the 10 En% casein diet (–0.07 ± 0.03 MJ/d; P < 0.05); carbohydrate and fat balances did not differ between the treatments. EE did not differ when participants consumed the 25 En% or 10 En% diets. Participants were in higher protein balance (0.56 ± 0.05 vs. 0.30 ± 0.04 MJ/d; P < 0.0001), lower carbohydrate balance (0.86 ± 0.14 vs. 1.37 ± 0.17 MJ/d; P < 0.01), and similar negative fat balance when they consumed the 25 En% casein compared with the 25 En% gelatin diet. In conclusion, when we compared the effects of an incomplete protein (gelatin) and a complete protein (casein) at 2 concentrations over 36 h, gelatin resulted in a greater appetite suppression; casein caused a greater positive (smaller negative) protein balance, and effects on EE did not differ. In terms of weight loss for people with obesity, the greater hunger-suppressing effect of gelatin may play a role in reducing energy intake if this effect is maintained when consuming a gelatin diet in the long term. In addition, long-term use of casein may contribute to preservation of fat-free mass.

Cosupplementation of Isoflavones, Prenylflavonoids, and Lignans Alters Human Exposure to Phytoestrogen-Derived 17{beta}-Estradiol Equivalents [Nutrient Physiology, Metabolism, and Nutrient-Nutrient Interactions]

Fri, 20 Nov 2009 10:01:59 PST

The microbial metabolism of dietary phytoestrogens varies considerably among individuals and influences the final exposure to bioactive compounds. In view of the increasing number of food supplements combining several classes of phytoestrogens, the microbial potential to activate various proestrogens within an individual was evaluated in 3 randomized dietary crossovers. Treatment allocation was based on participants' eligibility (>45% in vitro bioactivation of ≥2 separate proestrogens by fecal cultures; n = 40/100). After a run-in of ≥4 d, participants were given soy-, hop-, and/or flax-based food supplements dosed either separately (SOY: 2.83 mg daidzein aglycone equivalents/supplement, HOP: 1.20 mg isoxanthohumol (IX)/supplement, or FLAX: 2.08 mg secoisolariciresinol (SECO) aglycone equivalents/supplement; reference intervention) or simultaneously (MIX; test intervention) 3 times/d for 5 d, followed by a wash-out period (≥7 d) and the second intervention. Before and after each (co)supplementation, spot urine and serum were collected. In total, 22 equol, 19 8-prenylnaringenin (8-PN), and 21 enterolactone (ENL) producers completed the SOY+MIX, HOP+MIX, and FLAX+MIX trials, respectively. The microbial bioactivation of daidzein, IX, and SECO, generally decreased upon coincubation in vitro (equol: 4.4%, P = 0.164; 8-PN: 20.5%, P < 0.001; ENL: 44.3%, P < 0.001) and cosupplementation in vivo (equol: 28.3%, P = 0.009; 8-PN: 35.4%, P = 0.107; ENL: 35.9%, P = 0.003). Although the bioavailabilities of total isoflavones, prenylflavonoids, and lignans were not significantly affected upon coadministration, participants were exposed to lower phytoestrogen-derived 17β-estradiol equivalents. In conclusion, the bioavailability of phytoestrogens, especially when given in mixtures, is subject to high interindividual variation. These findings support the importance of personalized screening when assessing the efficacy of such products and mixtures.

Apolipoprotein A5 Polymorphisms Interact with Total Dietary Fat Intake in Association with Markers of Metabolic Syndrome in Puerto Rican Older Adults [Genomics, Proteomics, and Metabolomics]

Mattei, J., Demissie, S., Tucker, K. L., Ordovas, J. M. — Fri, 20 Nov 2009 10:01:59 PST

APOA5 -1131T > C and S19W single nucleotide polymorphisms (SNP) have been consistently associated with plasma lipid concentration and metabolic syndrome (MetS), alone and in modulation by dietary factors. Puerto Ricans have a high prevalence of metabolic conditions and high minor allele frequency for these SNP, suggesting a possible role in disease for this population. We aimed to determine the association of APOA5 -1131T > C and S19W with plasma lipids and markers of MetS, alone and in interaction with total fat intake, as a percent of total energy intake, in Puerto Ricans. Anthropometric and demographic data, FFQ, and blood samples were collected at baseline from participants in the Boston Puerto Rican Health Study (n = 802, 45–75 y). APOA5 S19W was associated with plasma HDL cholesterol (HDL-C) (P = 0.044); minor allele carriers had lower HDL-C [1.12 ± 0.03 (mean ± SE)] than those with the common variant (1.18 ± 0.01 mmol/L), even after adjustment for plasma triglycerides (TG) (P = 0.012). Neither polymorphism was associated with TG or other lipids. Interaction of the -1131T > C SNP with total fat energy intake was observed for plasma TG (P = 0.032) and total cholesterol (P = 0.034). APOA5 S19W interacted with total fat intake in association with systolic (P = 0.002) and diastolic (P = 0.007) blood pressure. Neither SNP was associated with MetS in the overall analysis or after stratifying by total energy intake as fat. In conclusion, Puerto Ricans present a distinctive lipid profile in association with APOA5 polymorphisms. Dietary fat intake seems to modulate these associations. The results contribute to the understanding of health disparities in this population.

A Metabolite Profiling Approach to Identify Biomarkers of Flavonoid Intake in Humans [Genomics, Proteomics, and Metabolomics]

Fri, 20 Nov 2009 10:01:59 PST

Flavonoids are phytochemicals that are widespread in the human diet. Despite limitations in their bioavailability, experimental and epidemiological data suggest health benefits of flavonoid consumption. Valid biomarkers of flavonoid intake may be useful for estimating exposure in a range of settings. However, to date, few useful flavonoid biomarkers have been identified. In this study, we used a metabolite profiling approach to examine the aromatic and phenolic profile of plasma and urine of healthy men after oral consumption of 200 mg of the pure flavonoids, quercetin, (-)-epicatechin, and epigallocatechin gallate, which represent major flavonoid constituents in the diet. Following enzymatic hydrolysis, 71 aromatic compounds were quantified in plasma and urine at 2 and 5 h, respectively, after flavonoid ingestion. Plasma concentrations of different aromatic compounds ranged widely, from 0.01 to 10 µmol/L, with variation among volunteers. None of the aromatic compounds was significantly elevated in plasma 2 h after consumption of either flavonoid compared with water placebo. This indicates that flavonoid-derived aromatic compounds are not responsible for the acute physiological effects reported within 2 h in previous human intervention studies involving flavonoids or flavonoid-rich food consumption. These effects are more likely due to absorption of the intact flavonoid. Our urine analysis suggested that urinary 4-ethylphenol, benzoic acid, and 4-ethylbenzoic acid may be potential biomarkers of quercetin intake and 1,3,5-trimethoxybenzene, 4-O-methylgallic acid, 3-O-methylgallic acid, and gallic acid may be potential markers of epigallocatechin gallate intake. Potential biomarkers of (-)-epicatechin were not identified. These urinary biomarkers may provide an accurate indication of flavonoid exposure.

Body Mass Index Is an Important Determinant of Methylation Biomarkers in Women of Reproductive Ages [Nutritional Epidemiology]

Fri, 20 Nov 2009 10:01:59 PST

B vitamin deficiencies lead to moderate hyperhomocysteinemia, which has been associated with health and disease. However, concomitant derangements in cellular methylation, reflected by altered plasma S-adenosylmethionine (SAM) or S-adenosylhomocysteine (SAH) concentrations, may be the primary cause. Therefore, we identified determinants of homocysteine, SAM, and SAH concentrations in 336 women, aged 20–48 y, as part of a large study focusing on risk factors for reproductive disorders. Blood was obtained to determine plasma SAM, SAH, and total homocysteine (tHcy), serum vitamin B-12 and folate, RBC folate concentrations, and the related single nucleotide polymorphisms 5,10-methylenetetrahydrofolate reductase (MTHFR) 677C > T and 1298A > C, methionine synthase reductase (MTRR) 66A > G, and nicotinamide N-methyltransferase IVS1–151G > A. Questionnaires provided information on demographics, lifestyles, and nutrient intakes. Correlation coefficients were calculated and multivariable associations were assessed with a general linear model. Serum folate was positively correlated with SAM concentrations (r = 0.159; P = 0.004). Folate and vitamin B-12 were not correlated with SAH concentrations or the SAM:SAH ratio but were inversely correlated with tHcy concentrations (serum folate r = –0.324; RBC folate r = –0.294; vitamin B-12 r = –0.307; P < 0.01). From the multivariable analysis, BMI was the strongest determinant of SAM (standardized β = 19.145; P < 0.001) and SAH concentrations (standardized β = 3.241; P = 0.010). MTHFR 677TT (standardized β = 0.195; P = 0.001), B vitamin supplement use (standardized β = –0.156; P < 0.001) and dietary protein intake (standardized β = –0.011; P < 0.001) were the strongest determinants of tHcy concentrations. Thus, the determinants of SAM and SAH differ from those of tHcy concentrations. Given that BMI was a strong determinant of SAM concentrations, it should be included in future studies on cellular methylation.

Dietary Patterns are Linked to Cardiovascular Risk Factors but Not to Inflammatory Markers in Alaska Eskimos [Nutritional Epidemiology]

Fri, 20 Nov 2009 10:01:59 PST

Despite the tradition of a diet high in fish oils and abundant physical activity, coronary artery disease is increasing among Alaska Eskimos. Explanations for this observation include lifestyle changes. In this cross-sectional analysis, we evaluated dietary patterns of Alaska Eskimos and investigated the relations between these dietary patterns and known cardiovascular risk factors, including inflammatory markers. We used a principal component analysis with data from FFQ collected in 2000–2004 to determine dietary patterns of Alaska Eskimos. Four dietary patterns were identified: a traditional pattern, plus 3 patterns based on purchased food, one of which reflected healthy food choices. The traditional dietary pattern was associated with lower triglycerides (P < 0.001) and blood pressure (P = 0.04) and slightly higher LDL cholesterol (LDL-C) (P = 0.05). Whereas the healthy purchased diet was associated with a trend toward lower LDL-C (P = 0.09), the beverages and sweets diet was positively associated with LDL-C (P = 0.02). Diet pattern was not associated with inflammatory markers or pathogen burden. Our data show that the traditional diet is related to a better profile of cardiovascular disease risk factors and should be encouraged. Programs are needed to encourage the availability of healthy food choices for those not able to obtain traditional foods.

Intakes of (n-3) Fatty Acids and Fatty Fish Are Not Associated with Cognitive Performance and 6-Year Cognitive Change in Men Participating in the Veterans Affairs Normative Aging Study [Nutritional Epidemiology]

Fri, 20 Nov 2009 10:01:59 PST

High intake of fish and (n-3) PUFA may protect against age-related cognitive decline. However, results are inconsistent and limited data exist regarding changes in multiple cognitive functions over a longer period of time. In this study, we assessed the association between fatty fish intake as well as (n-3) PUFA intake with cognitive performance and cognitive change over 6 y in 1025 elderly men. Participants were from the Veterans Affairs Normative Aging Study. Cognitive function was assessed with a battery of cognitive tests focusing on factors representing memory/language, speed, and visuospatial/attention. Dietary intakes were assessed with a validated FFQ. We used general linear models to assess cross-sectional associations and mixed models to assess the associations over time. Models were adjusted for age, education, BMI, smoking, diabetes, and intake of alcohol, saturated fat, vitamin C, and vitamin E. The mean age of participating men was 68 y at baseline. Median fish consumption ranged from 0.2 to 4.2 servings/wk across quartiles. Cross-sectional analyses showed no association between fatty fish or (n-3) PUFA intake and cognitive performance. Longitudinal analyses, over 6 y of follow-up, also did not show any significant associations between fatty fish or (n-3) PUFA intake and cognitive change. In this population of elderly men, intake of neither fatty fish nor (n-3) PUFA was associated with cognitive performance.

Past and Current Body Size Affect Validity of Reported Energy Intake among Middle-Aged Danish Men [Nutritional Epidemiology]

Fri, 20 Nov 2009 10:01:59 PST

Our objectives were to estimate the degree of misreporting energy intake (EI) and analyze associations with previous BMI, current BMI, or both. The study was part of the Adiposity and Genetics Study follow-up study including 309 Danish men (age 40–65 y) originally sampled from the obligatory draft board examination. Height and weight were measured at the mean ages of 20 (draft board), 33, 44, and 49 y (current age). Obesity was categorized as BMI ≥ 31 kg/m². Dietary intake for 7 d and physical activity (PA) level (PAL) were self-reported. Resting metabolic rate (RMR) was measured in a ventilated hood system. By comparing EI with energy expenditure and assuming energy balance, reporting accuracy (RA) was estimated as EI/(RMR·PAL). A plausibility interval was calculated to encompass specific variation components of EI, RMR, and PAL; the specific 95% plausibility interval was 1.00 ± 0.35. Participants were categorized as underreporters (RA ≤ 0.65), plausible reporters (0.65 < RA ≤ 1.35), or overreporters (RA > 1.35) of EI. The relation between RA and BMI was studied through linear regression analysis. Overall, the RA was (mean ± SE) 0.76 ± 0.01. Of 309 participants, 35% underreported and 7% overreported. Whether stratified for current BMI or draft board BMI, the obese men were more likely to underreport than those who were not obese. Among those currently not obese, underreporting was more prevalent among those who were obese at the draft board examination (44%) than among those who were not (21%). Regression analysis showed that both previous and current BMI and their combination were significantly associated with RA. Thus, underreporting of dietary intake seems to be associated with not only current BMI but also with current BMI in combination with previous BMI.

Dietary Arachidonic Acid to EPA and DHA Balance Is Increased among Canadian Pregnant Women with Low Fish Intake [Nutritional Epidemiology]

Friesen, R. W., Innis, S. M. — Fri, 20 Nov 2009 10:01:59 PST

Arachidonic [ARA, 20:4(n-6)], eicosapentaenoic [EPA, 20:5(n-3)], and docosahexaenoic acids [DHA, 22:6(n-3)] occur in the diet in animal tissue lipids, play important roles in human development and health, but have interactive and opposing functions. Meat and poultry have higher ARA and fish are richer in EPA and DHA. National databases were recently revised to include complete data on ARA in foods. We used a validated FFQ and the revised nutrient databases to quantify the distribution of ARA, EPA, and DHA intakes and balance for 204 healthy Canadian pregnant women. We focused on intake distributions because risk of adverse health effects increases at lower nutrient intakes. RBC fatty acids were analyzed concurrenly with dietary assessment. The distribution of ARA, EPA, and DHA intakes were skewed (P < 0.001), with a median (5–95th percentile) of 107 (41–225), 65 (10–228), and 105 (10–430) mg/d ARA, EPA, and DHA, respectively. Fish provided 66 and 76% of EPA and DHA, respectively, whereas eggs, poultry, and meats provided 81% of ARA. Women consuming <101 g fish/wk consumed less EPA and DHA and had markedly elevated median dietary ARA:EPA and ARA:DHA ratios and RBC lipid ARA:EPA + DHA ratios compared with women consuming ≥101 g fish/wk (P < 0.001). Relatively small increases in fish intake of 1–2 servings (25–50 g)/wk corrected the distorted dietary (n-6):(n-3) fatty acid balance among women consuming meats, but not fish. Median fish and DHA intakes below the recommended 1–2 servings/wk fish for pregnant women suggest major changes in the availability, cost, or acceptance of fish are needed.

A Conjugated Linoleic Acid-Enriched Beef Diet Attenuates Lipopolysaccharide-Induced Inflammation in Mice in Part through PPAR{gamma}-Mediated Suppression of Toll-Like Receptor 4 [Nutritional Immunology]

Fri, 20 Nov 2009 10:01:59 PST

Conjugated linoleic acid (CLA) is a PUFA found in beef and dairy products that has immunoregulatory properties. The level of CLA in beef can be enhanced by feeding cattle fresh grass rather than concentrates. This study determined the effect of feeding a high-CLA beef diet on inflammation in an in vivo model of septic shock. Mice were fed a high-CLA beef (4.3% total fatty acid composition) or low-CLA beef diet (0.84% total fatty acid composition) for 6 wk. Lipopolysaccharide (LPS; 3 µg) or sterile PBS was injected i.v. and serum was harvested 6 h after injection. Serum interleukin (IL)-1β, IL-12p70, IL-12p40, and interferon- concentrations were significantly reduced in response to the LPS challenge in the high-CLA beef diet group. Bone marrow-derived dendritic cells (BMDC) from the high-CLA beef diet group had significantly less IL-12 and more IL-10 in response to ex vivo LPS stimulation. Furthermore, toll-like receptor 4 (TLR4) and CD14 protein and mRNA expression on BMDC was significantly attenuated in the high-CLA compared with the low-CLA beef diet group. Complimentary in vitro experiments to determine the specificity of the effect showed that synthetic cis9, trans11-CLA suppressed surface expression of CD14 and TLR4 on BMDC. Treatment with the PPAR inhibitor GW9662 partially reversed TLR4 expression in immature BMDC. The results of this study demonstrate that feeding a diet enriched in high-beef CLA exerts profound antiinflammatory effects in vivo within the context of LPS-induced sepsis. In addition, downregulation of BMDC TLR4 is mediated through induction of PPAR.

Increased Tissue Arachidonic Acid and Reduced Linoleic Acid in a Mouse Model of Cystic Fibrosis Are Reversed by Supplemental Glycerophospholipids Enriched in Docosahexaenoic Acid [Nutrition and Disease]

Fri, 20 Nov 2009 10:01:59 PST

An imbalance in (n-6)/(n-3) PUFA has been reported in cystic fibrosis (CF) patients. Glycerophospholipids enriched in docosahexaenoic acid (GPL-DHA) have been shown to regulate the (n-6)/(n-3) fatty acid ratio in the elderly. Here, we tested the effect of GPL-DHA supplementation on PUFA status in F508del homozygous CF mice. GPL-DHA liposomes were administrated by gavage (60 mg DHA/kg daily, i.e. at maximum 1.4 mg DHA/d) to 1.5-mo-old CF mice (CF+DHA) and their corresponding wild-type (WT) homozygous littermates (WT+DHA) for 6 wk. The PUFA status of different tissues was determined by GC and compared with control groups (CF and WT). There was an alteration in the (n-6) PUFA pathway in several CF-target organs in CF compared with WT mice, as evidenced by a higher level of arachidonic acid (AA) in membrane phospholipids or whole tissue (21 and 39% in duodenum-jejunum, 32 and 38% in ileum, and 19 and 43% in pancreas). Elevated AA levels were associated with lower linoleic acid (LA) and higher dihomo--linolenic acid levels. No DHA deficiency was observed. GPL-DHA treatment resulted in different PUFA composition changes depending on the tissue (increase in LA, decrease in elevated AA, DHA increase, increase in (n-6)/(n-3) fatty acid ratio). However, the DHA/AA ratio consistently increased in all tissues in CF+DHA and WT+DHA mice. Our study demonstrates the effectiveness of an original oral DHA formulation in counter-balancing the abnormal (n-6) fatty acid metabolism in organs of CF mice when administrated at a low dose and highlights the potential of the use of GPL-DHA as nutritherapy for CF patients.

A New Dietary Inflammatory Index Predicts Interval Changes in Serum High-Sensitivity C-Reactive Protein [Nutrition and Disease]

Fri, 20 Nov 2009 10:02:00 PST

Inflammation is associated with a number of chronic conditions, such as cancer and cardiovascular disease. Reducing inflammation may help prevent or treat these conditions. Diet has consistently been shown to modulate inflammation. To facilitate research into the inflammatory effect of diet on health in humans, we sought to develop and validate an Inflammatory Index designed to assess the inflammatory potential of individuals' diets. An Inflammatory Index was developed based on the results of an extensive literature search. Using data from a longitudinal observational study that carefully measured diet and the inflammatory marker, serum high-sensitivity (hs) C-reactive protein (CRP), in ~600 adults for 1 y, we conducted analyses to test the effect of Inflammatory Index score on hs-CRP as a continuous and dichotomous (≤3 mg/L, >3 mg/L) indicator of inflammatory response, while controlling for important potential confounders. Results based on continuous measures of hs-CRP suggested that an increasing Inflammatory Index score (representing movement toward an antiinflammatory diet) was associated with a decrease in hs-CRP. Analyses using hs-CRP as a dichotomous variable showed that an antiinflammatory diet was associated with a decrease in the odds of an elevated hs-CRP (P = 0.049). The results are consistent with the ability of the Inflammatory Index to predict hs-CRP and provide additional evidence that diet plays a role in the regulation of inflammation, even after careful control of a wide variety of potential confounders.

Oral Administration of 3,3'-Diindolylmethane Inhibits Lung Metastasis of 4T1 Murine Mammary Carcinoma Cells in BALB/c Mice [Nutrition and Disease]

Fri, 20 Nov 2009 10:02:00 PST

3,3'-diindolylmethane (DIM) is the major in vivo product of the acid-catalyzed oligomerization of indole-3-carbinol present in cruciferous vegetables, and it has been shown to exhibit anticancer properties. In this study, we assessed the effects of DIM on the metastasis of 4T1 mouse mammary carcinoma cells. In vitro culture studies showed that DIM dose-dependently inhibited the migration, invasion, and adhesion of 4T1 cells at concentrations of 0–10 µmol/L without attendant changes in cell viability. In an in vivo lung metastasis model, 4T1 cells (2 x 10⁵ cells/mouse) were injected into the tail veins of syngeneic female BALB/c mice. Beginning on the second day, the mice were subjected to gavage with 0–10 mg DIM/(kg body weight · d) for 13 d. Oral DIM administration resulted in a marked reduction in the number of pulmonary tumor nodules. DIM treatment significantly reduced the levels of matrix metalloproteinase (MMP)-2, MMP-9, tissue inhibitor of metalloproteinase (TIMP)-1, and vascular cell adhesion molecule (VCAM)-1 and increased TIMP-2 levels in the sera and lungs of mice injected with 4T1 cells. Additionally, DIM treatment reduced the serum concentrations of interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF). We have demonstrated that DIM profoundly inhibits the lung metastasis of 4T1 cells, which was accompanied by reduced levels of MMP, adhesion molecules, and proinflammatory cytokines. These results indicate that DIM has potential as an antimetastatic agent for the treatment of breast cancer.

Dietary Fish Oil Exerts Hypolipidemic Effects in Lean and Insulin Sensitizing Effects in Obese LDLR-/- Mice [Nutrition and Disease]

Saraswathi, V., Morrow, J. D., Hasty, A. H. — Fri, 20 Nov 2009 10:02:00 PST

Obesity is often associated with dyslipidemia, insulin resistance, and hypertension. Together, these metabolic perturbations greatly increase the risk of developing cardiovascular disease and diabetes. Although fish oil is a well-established hypolipidemic agent, the mechanisms by which it mediates its lipid-lowering effects are not clear. In addition, it has not been established whether dietary fish oil has different effects in lean and obese mice. LDL receptor deficient (LDLR–/–) and leptin deficient mice on a LDLR–/– background (ob/ob;LDLR–/–) were fed a high fat diet (39% total fat) supplemented with 6% olive oil or fish oil for 6 wk. Fish oil supplementation resulted in lower concentrations of plasma total cholesterol (P < 0.01), triglycerides (P < 0.01), and free fatty acids (P < 0.001) in lean LDLR–/– mice, but not in ob/ob;LDLR–/– mice. In contrast, a fish oil diet did not modulate insulin sensitivity in lean LDLR–/– mice, but it improved insulin sensitivity in ob/ob;LDLR–/– mice (P < 0.05) compared with olive oil fed ob/ob;LDLR–/– mice. Interestingly, plasma adiponectin concentrations were significantly higher and hepatic steatosis was reduced in both mouse models upon fish oil feeding. Finally, fish oil fed LDLR–/– mice exhibited higher hepatic AMP activated protein kinase (AMPK) phosphorylation (P < 0.05), whereas AMPK phosphorylation was not elevated by fish oil feeding in ob/ob;LDLR–/– mice. Taken together, our data suggest that fish oil reduces hepatic steatosis in both lean and obese mice, has potent plasma lipid lowering effects in lean mice, and exerts insulin sensitizing effects in obese mice.

Overview to Symposium "Nutrients and Epigenetic Regulation of Gene Expression" [Symposium: Nutrients and Epigenetic Regulation of Gene Expression]

Ho, E., Zempleni, J. — Fri, 20 Nov 2009 10:02:00 PST

The American Society for Nutrition hosted a symposium entitled Nutrients and Epigenetic Regulation of Gene Expression at the Experimental Biology meeting on April 20, 2009, in New Orleans, LA. The symposium was cochaired by Emily Ho from Oregon State University and the Linus Pauling Institute, and Janos Zempleni from the University of Nebraska at Lincoln. The goal of this symposium was to highlight the interactions among nutrients, epigenetics, and disease susceptibility. The symposium featured 4 speakers, each presenting novel insights into mechanisms by which nutrients participate in gene regulation. Janos Zempleni elucidated mechanisms by which the covalent binding of biotin to histones represses transposable elements, thereby enhancing genome stability. Emily Ho shared valuable insights into bioactive food compounds that inhibit histone deacetylases. James Kirkland from the University of Guelph in Canada talked about a niacin-dependent poly(ADP-ribosylation) of histones, an epigenetic mark that is not currently being given full consideration in nutrition. Patrick Stover from Cornell University described the interrelationships among 1-carbon metabolism, DNA methylation, gene silencing, and their influence in the etiology of folate-related pathologies. All 4 presentations were videotaped and can be viewed online (www.nutrition.org).

Repression of Transposable Elements by Histone Biotinylation [Symposium: Nutrients and Epigenetic Regulation of Gene Expression]

Zempleni, J., Chew, Y. C., Bao, B., Pestinger, V., Wijeratne, S. S. K. — Fri, 20 Nov 2009 10:02:00 PST

Transposable elements constitute >40% of the human genome; transposition of these elements increases genome instability and cancer risk. Epigenetic mechanisms are important for transcriptional repression of retrotransposons, thereby preventing transposition events. Binding of biotin to histones, mediated by holocarboxylase synthetase (HCS), is a novel histone mark that plays a role in gene regulation. Here, we review recent findings that biotinylation of lysine-12 in histone H4 (H4K12bio) is an epigenetic mechanism to repress long terminal repeat (LTR) retrotransposons in human and mouse cell lines, primary cells from human adults, and in Drosophila melanogaster. Further, evidence is summarized that supports a causal relationship between the repression of LTR in H4K12bio-depleted cells and increased production of viral particles, increased frequency of retrotransposition events, and increased frequency of chromosomal abnormalities in mammals and Drosophila. Although HCS interacts physically with histones H3 and H4, the mechanism responsible for targeting HCS to retrotransposons to mediate histone biotinylation is uncertain. We hypothesize that HCS binds specifically to genomic regions rich in methylated cytosines and catalyzes increased biotinylation of histone H4 at lysine-12. Further, we hypothesize that this biotinylation promotes the subsequent dimethylation of lysine-9 in histone H3, resulting in an overall synergistic effect of 3 diet-dependent covalent modifications of histones in the repression of LTR.

Dietary Sulforaphane, a Histone Deacetylase Inhibitor for Cancer Prevention [Symposium: Nutrients and Epigenetic Regulation of Gene Expression]

Ho, E., Clarke, J. D., Dashwood, R. H. — Fri, 20 Nov 2009 10:02:00 PST

The reversible acetylation of histones is an important mechanism of gene regulation. During prostate cancer progression, specific modifications in acetylation patterns on histones are apparent. Targeting the epigenome, including the use of histone deacetylase (HDAC) inhibitors, is a novel strategy for cancer chemoprevention. Recently, drugs classified as HDAC inhibitors have shown promise in cancer clinical trials. We have previously found that sulforaphane (SFN), a compound found in cruciferous vegetables, inhibits HDAC activity in human colorectal and prostate cancer cells. Based on the similarity of SFN metabolites and other phytochemicals to known HDAC inhibitors, we previously demonstrated that sulforaphane acted as an HDAC inhibitor in the prostate, causing enhanced histone acetylation, derepression of P21 and Bax, and induction of cell cycle arrest/apoptosis, leading to cancer prevention. The ability of SFN to target aberrant acetylation patterns, in addition to effects on phase 2 enzymes, may make it an effective chemoprevention agent. These studies are important because of the potential to qualify or change recommendations for high-risk prostate cancer patients and thereby increase their survival through simple dietary choices incorporating easily accessible foods into their diets. These studies also will provide a strong scientific foundation for future large-scale human clinical intervention studies.

Niacin Status Impacts Chromatin Structure [Symposium: Nutrients and Epigenetic Regulation of Gene Expression]

Kirkland, J. B. — Fri, 20 Nov 2009 10:02:00 PST

Niacin is required to form NAD and NADP, which are involved in many essential redox reactions in cellular metabolism. In addition, NAD⁺ acts as a substrate for a variety of ADP-ribosylation reactions, including poly- and mono-ADP-ribosylation of proteins, formation of cyclic ADP-ribose, and the generation of O-acetyl-ADP-ribose in deacetylation reactions. These nonredox reactions are critical in the regulation of cellular metabolism, and they are sensitive to dietary niacin status. There are 4 known mechanisms by which ADP-ribosylation reactions directly regulate chromatin structure. These include the covalent modification of histones with poly(ADP-ribose), the extraction of histones from chromatin by noncovalent binding to poly(ADP-ribose) on poly(ADP-ribose) polymerase-1, poly ADP-ribosylation of telomeric repeat-binding factor-1 within telomeres, and deacetylation of histones by the sirtuins. These reactions produce a variety of localized effects in chromatin structure, and altered function in response to changes in niacin status may have dramatic effects on genomic stability, cell division and differentiation, and apoptosis.

One-Carbon Metabolism-Genome Interactions in Folate-Associated Pathologies [Symposium: Nutrients and Epigenetic Regulation of Gene Expression]

Stover, P. J. — Fri, 20 Nov 2009 10:02:00 PST

Impairments in folate-mediated 1-carbon metabolism are associated with several common diseases and developmental anomalies including intestinal cancers, vascular disease, cognitive decline, and neural tube defects. The etiology of folate-associated pathologies involves interactions among multiple genetic risk alleles and environmental factors, although the causal mechanisms that define the role of folate and other B-vitamins in these complex disorders remain to be established. Folate and other B-vitamins fundamentally differ from other nutrients that interact with the genome in determining health and disease outcomes in that their interaction is reciprocal. Common gene variants influence the activity of folate-dependent enzymes and anabolic pathways; folate-mediated 1-carbon metabolism is essential for the high-fidelity synthesis of DNA and activated methyl groups that are required for DNA methylation and regulation of chromatin structure. This review focuses on the regulation of folate-mediated 1-carbon metabolism and its role in maintaining genome integrity and on strategies for establishing the metabolic pathways and mechanisms that underlie folate-associated pathologies.

Announcements [Announcements]

Fri, 20 Nov 2009 10:02:00 PST