Energy and Macronutrient Metabolism

CURRENT DEVELOPMENTS IN NUTRITIONFROM THE AMER ICANSOCIETY FOR NUTR IT ION

© 2018 American Society for Nutrition. All rightsreserved.

Manuscript received September 20, 2017. Initialreview completed March 23, 2018. Revision acceptedMay 27, 2018. Published online July 4, 2018.

Energy and Macronutrient Metabolism

Metabolic Flexibility in Overweight or Obese Women Consuming Controlled Diets (E06-01)

Nancy L Keim,1 Justin Waller,1 Sridevi Krishnan,2 William F Horn,1 Remy Bosivel,2and John Newman1

1USDA,ARS,WesternHumanNutrition ResearchCenter; and 2University of California-Davis

Objective: The primary aim was to determine if consumption of specific diet patterns affectedmetabolic flexibility, which was evaluated by a high-fat meal challenge. A secondary aim wasto determine if the amount of fat combusted in response to the meal challenge was related tocirculating lipids.

Methods: An 8-wk, randomized trial with controlled food intake was conducted withoverweight or obese women. Subjects were given diets based on the Dietary Guidelines forAmericans (DGA, n = 22) or a more typical American diet (TAD, n = 22). A distinguishing dietfeature was the polyunsaturated fatty acid+monounsaturated fatty acid-to-saturated fat ratio (2.5forDGA, 1.4 for TAD).Ahigh-fatmeal challengewas administered atweeks 0 and 8,with 800 kcal,60% fat, 25% carbohydrate, and 15%protein. Pre- and postmeal (45, 180, and 360min) respiratorygases and blood samples were taken to determine fat combustion rates and blood lipids.

Results: There were no significant diet- or week-related differences in the quantity of fatcombusted over the 6-h postprandial period of the meal challenge. Mean ± SEM values for DGAat weeks 0 and 8 were 38% ± 2% and 38% ± 3% of the dietary fat load; and TAD were 37% ± 4%and 37% ± 3% at weeks 0 and 8. Stability of the fat combustion response between weeks wasevaluated within subjects, regardless of diet assignment: 17 had consistently higher responses,burning 51% ± 3% and 49% ± 3% of the fat load at weeks 0 and 8, whereas 17 others were foundto have lower responses, burning 26% ± 2% and 2%5 ± 2% of the fat load at weeks 0 and 8.The lower fat combustion group had higher postprandial triglycerides at 180 and 360 min afterthe meal compared with the higher fat combustion group (P < 0.04). Values were 304 ± 25 vs.210± 20 mg/dL at 180 min and 298± 27 vs. 198± 23 mg/dL at 360 min for the lower and higherfat combustion groups, respectively. Finally, the group with lower fat combustion was older thanthe higher fat combustion group, at 53 ± 2 vs. 46 ± 3 y, respectively (P < 0.02).

Conclusions: Fat combusted in response to a high-fat test meal is stable over time undercontrolled diet conditions and is likely not related to the type of fat in the diet. Whenchallenged with a high-fat meal, a lower fat combustion response is related to higher postprandialtriglyceridemia.

Funding SourcesUSDA CRIS 2032-51,530-022, the National Dairy Council, and the Campbell Soup Company.

Metabolic Networks Linking Red Meat and Coffee Consumption to Type 2 DiabetesIncidence (E06-02)

ClemensWittenbecher,1 JanKrumsiek,2 Cornelia Prehn,3 JerzyAdamski,3 Tobias Pischon,4Heiner Boeing,1 and Matthias B Schulze1

1German Institute of Human Nutrition Potsdam-Rehbruecke (DIfE); 2Institute of Com-putational Biology (ICB), Helmholtz-Zentrum München, Neuherberg, Germany; 3Helmholtz-ZentrumMuenchen, Neuherberg, Germany; and 4Max-Delbrück Center for Molecular Medicine(MDC), Berlin, Germany

Objective: The aim of the study was to determine if there were any links between red meat orcoffee consumption and type 2 diabetes (T2D) incidence.

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Methods: Working with the EPIC-Potsdam cohort (n = 27,548),we applied a nested case-cohort design (subcohort = 2092, incidentT2D cases = 692) with baseline information on targeted serummetabolomics (126 amino acids, acylcarnitines, and glycerophospho-lipids), and self-assessed dietary habits. We developed the causalinference-based NETcoupler algorithm, which links diet and T2D todata-drivenMNs. For eachmetabolite, comprehensively adjusted linearregression and Cox models were used to evaluate links of MNs withdiet and with time to T2D, respectively. Further adjustment for directMN neighbors controlled confounding and indirect effects throughother metabolites. From the resulting joint diagrams, metabolites thatlinked diet to T2D risk were selected for evaluation in mediationanalyses.

Results: Red meat consumption was associated with higher T2Drisk (HR: 1.25; 95% CI: 1.00, 1.55). From the joint graphical models,sphingomyelin (SM) C18:0, the alkyl-acyl phosphatidylcholines (PCs)C36/0 and C36/4 (all linked to high redmeat intake and high T2D risk),

FIGURE 1 Potential metabolic links between habitual consumption of red meat and coffee, and type 2 diabetes risk.

and glycine (linked to low red meat intake and low T2D risk) wereconsidered as potential mediators (Figure 1). Controlling for thesemetabolic markers attenuated the red meat–related T2D risk by 70%(95% CI: 43%, 100%). Coffee intake was associated with lower T2D risk(HR 0.76, 95% CI: 0.62, 0.94). From the networks, SM C20:2, lysoPCC18:2, and alkyl-acyl PC C34/3 (all linked to high coffee intake and lowT2D risk), and phenylalanine and lysoPC C14:0 (linked to low coffeeintake and high T2D risk), were selected as intermediates. Controllingfor these potential mediators attenuated the coffee-related T2D risk by66% (95% CI: 38%, 100%).

Conclusions: We have developed an algorithm that generates jointnetwork models of dietary exposures, MNs, and T2D risk. Adjustmentfor interlinking metabolites markedly attenuated the diet-related dis-ease risk, which is consistent with the hypothesis of the tagged pathwaysplaying a major role in relating the investigated foods to T2D risk.

Funding SourcesGermanCenter of Diabetes Research (DZD),München-Neuherberg

CURRENT DEVELOPMENTS IN NUTRITION

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Energy and Macronutrient Metabolism 3

A Critical Role for the Antidiuretic Hormone Vasopressin inFructose-Induced Metabolic Syndrome (E06-03)

Miguel A Lanaspa, Thomas Jensen, Masanari Kuwabara, AnaAndres-Hernando, and Richard Johnson

University of Colorado AMC

Objectives: For a long time, vasopressin has been thought tobe a hormone that is just involved in water retention and urinaryconcentration. However, it has recently been postulated that it alsopromotes extrarenal effects, including the storage of fat as a source ofmetabolic water. We and others have shown that fructose, a heavilyused sugar in the form of either sucrose or high-fructose corn syrup,induces important features of metabolic syndrome, including obesity,fatty liver, and insulin resistance. Recently, it has also been found thatfructose stimulates vasopressin release, thus leading us to the hypothesisthat in metabolic syndrome, chronic fructose ingestion might mediateitsmetabolic effects by activating vasopressin-dependent pathways. Theaim of this study was to test this hypothesis.

Methods: Male C57BL/6 wild-type (WT), Vasopressin 1a receptorKnockout (V1aKO), and Vasopressin 1b receptor Knockout (V1bKO)mice were maintained in either regular water or 15% fructose waterad libitum for 30 wk, and features of metabolic syndrome werecharacterized. We also analyzed copeptin concentrations, a surrogatefor vasopressin, in WT, V1aKO, V1bKO, fructokinase A knockout, andfructokinase A/C knockout mice on both water and 15% fructose waterfor 30 wk.

Results: Chronic fructose exposure resulted in significant elevationof plasma copeptin in all groups. However, despite similarly higherplasma vasopressin concentrations, V1bKO mice demonstrated signif-icant protection against fructose-induced weight gain, steatohepatitis,insulin resistance, and hyperleptinemia compared with wild-type mice.In contrast, V1aKO mice not only showed no sign of protectionagainst fructose but instead, showed a tendency for a worse metabolicphenotype in response to fructose. Furthermore, our data suggest thatthe mechanism whereby the blockade of the V1b receptor results insignificant protection against fructose-induced metabolic syndromeis mediated by the downregulation of fructokinase in the liver, thusleading to reduced dietary fructose metabolism.

Conclusion: To our knowledge, this is the first study to show thatvasopressin is elevated in mice chronically exposed to fructose, whereit plays an important deleterious role in fructose-induced metabolicsyndrome by activating an specific receptor V1b.

Funding SourcesNIH: R01 DK108859-03.

Plasma Oleoylethanolamide Concentrations Associate withGPR40 rs1573611 Variations in Participants from the Canola OilMulti-Centre Intervention Trial 2 (COMIT 2) (E06-04)

Jyoti Sihag,1 Dylan MacKay,2 Shatha S Hammad,1 Xiang Chen,1Kate J Bowen,4 Peter Eck,5 Benoît Lamarche,6 Todd C Rideout,7Philip W Connelly,8 Carla Taylor,3 Peter Zahradka,3 PatrickCouture,5 Sheila G West,10 Penny G Kris-Etherton,4 David JAJenkins,9 and Peter JH Jones1

1Richardson Centre for Functional Foods & Nutraceuticals, 2TheGeorge and Fay Yee Centre for Healthcare Innovation and 3CanadianCentre for Agri-food Research in Health and Medicine, St-Boniface

Hospital, University of Manitoba, Canada; 4The Pennsylvania StateUniversity; 5University ofManitoba, Canada; 6Institute ofNutrition andFunctional Foods, Laval University, Canada; 7School of Public Healthand Health Professions, University at Buffalo, NY; 8Keenan ResearchCentre for Biomedical Science and 9Li Ka Shing Knowledge Institute, StMichael’s Hospital, University of Toronto, Canada; and 10BiobehavioralHealth, The Pennsylvania State University

Background: Animal data suggest that oleoylethanolamide (OEA)acts as the agonist for G protein-coupled receptors (GPCRs). GPR40 isinvolved in fatty acid taste perception.

Objective: The aim of this study was to investigate plasma fattyacid ethanolamide (FAE) concentration in humans in response toconsumption of various dietary oils and to test for the association withthe GPR40 rs1573611 polymorphism.

Methods:Amulticenter, controlled-feeding, double-blind, random-ized, crossover trial was conducted, where participants (n = 115)consumed smoothies containing 20% kcal of total energy from higholeic canola oil (HOCO), regular canola oil (RCO), or control oil(CO) (an oil blend of butter, safflower, flaxseed, and coconut) twicea day for 6 wk. Plasma FAE concentrations were assessed usingultra-performance liquid chromatography-MS/MS. Genotyping wasperformed using quantitative polymerase chain reaction.

Results: Plasma OEA concentrations were the highest (P < 0.0001)with HOCO treatment (2.38 ± 0.25 ng/ml), followed by RCO(2.25 ± 0.25 ng/ml) and CO (1.92 ± 0.25 ng/ml) consumption. Inaddition, elevations (P < 0.0009) in plasma linoleoylethanolamide(0.68 ± 0.03 ng. ml–1) were observed after the CO treatment whencompared with the HOCO treatment (0.62 ± 0.03 ng.ml–1). Further-more, RCO and CO showed the highest (P < 0.0002) concentrationsof α-linoleoylethanolamide when compared with HOCO treatment.rs1573611 genotypes differentially affected arachidonoylethanolamideconcentrations (P < 0.0097). When diet-gene interactions were tested,elevated concentrations of plasma OEA were observed in participantspossessing the rs1573611-T allele (n = 48, P < 0.0037) after the con-sumption of HOCO (P < 0.0001) and RCO (P < 0.0121), but not CO.

Conclusions: Plasma OEA concentrations reflected the dietarypattern of oleic acid intake and may be influenced by GRP40 rs1573611polymorphism. Future trials should attempt to replicate the associationreported betweenOEA concentrations and rs1573611. Further researchis required to elucidate the role of OEA in human physiology.

This trial was registered at clinicaltrials.gov as NCT02029833.Funding SourcesThis research is part of the Canola Agri-Science Cluster, with

funding provided by Agriculture and Agri-Food Canada, the CanolaCouncil of Canada, Dow AgroSciences, Alberta Canola, SaskCanola,and the Manitoba Canola Growers.

Incremental Increases inDietary Carbohydrate Alter Plasma andCellular Unsaturated Fats Associated with Insulin Resistance inAdults with Metabolic Syndrome (E06-05)

Daniel J Freidenreich,1 Brittanie Volk,2 Laura J Kunces,2 Juan CArtistizabal,2 Catherine Saenz,2 William Kraemer,2 Stephen DPhinney,3 and Jeff S Volek2

1Rowan University, NJ; 2University of Connecticut; and 3Universityof California Davis


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Objectives: The aims were to investigate 1) how variations inthe dietary CHO: FAT ratio would alter the unsaturated fatty acidcomposition of plasma and buccal cell membrane lipids, and 2) if thesechanges, specifically the proportions of arachidonic acid (ARA) anddihomo-γ -linolenic acid (DGLA), were related to insulin resistance.

Methods: Individuals with metabolic syndrome (n = 15) werecontinuously fed six 3-wk hypocaloric diets,∼300 kcal below energy re-quirements, of individually prepared foods that progressively increasedin carbohydrates (from 47 to 346 g/d) (P1→P6) with concomitantdecreases in total fat. Each diet phase meal plan was generated usingnutrient analysis software with caloric intake individualized for eachparticipant. Saturated,monounsaturated, and polyunsaturated fats werekept at ∼40%, 33%, and 17% of total fat content, respectively, forevery phase. Protein was maintained at 1.8 g/kg reference weight. Thefollowing assessments were repeated at the beginning of the study andthe end of each diet phase: anthropometrics, body composition by dual-energy X-ray absorptiometry, buccal cell collection by cheek swab, andblood collection. Plasma and buccal cell samples were analyzed forfatty acid composition by high-resolution capillary gas chromatography.Plasma insulin and glucose were measured to assess insulin sensitivityusing the homeostasis model of assessment (HOMA2).

Results: Both buccal cell and plasma phospholipid ARA werehighest during P1 and lowest at P6. DGLA showed an inverse pattern toARA, peaking during P6 and at its lowest at P1. Palmitoleate (16:1n–7),a marker of de novo lipogenesis, was significantly lower in P1 than in P6in buccal cells, and in plasma triglyceride and cholesteryl esters. Plasmaphospholipid ARA was positively correlated with insulin sensitivity(P < 0.05) in all diet phases except P5, but no buccal cell fatty acidswere correlated with insulin sensitivity.

Conclusions: In adults with metabolic syndrome, very low carbo-hydrate intake was associated with a plasma and cellular fatty acidprofile consisting of high ARA and low 16:1n–7 and DGLA, which maycontribute to improved insulin sensitivity.

Funding SourcesTheNational Cattlemen’s Beef Association, the EggNutritionCenter

and the National Dairy Council.

Associations of Weight Status and Carbohydrate Intake withGestational Weight Gain (E06-06)

Makenzie L Callahan,1 Camille Schneider,1 Barabara Gower,1Patrick Catalano,2 and Paula Chandler-Laney1

1University of Alabama at Birmingham; and 2Center for Repro-ductive Health/MetroHealth Medical Center, Case Western ReserveUniversity, OH

Background: Carbohydrate (CHO) intake has been associated withgestational weight gain (GWG), but the direction of this associationis inconsistent. In the general population, some evidence suggests thatindividuals with obesity gainmore weight consuming a high-CHOdiet.

Objectives: The purpose of this study was to test the hypothesis thatwomen who are obese at entry to prenatal care have greater GWGwheneating a high- compared with low-CHO diet. We also hypothesized thatGWG would not differ by CHO for women of normal weight.

Methods:Healthywomenwith a BMI in the normal-weight (n= 31)compared with obese (n = 28) range enrolled in this study in earlypregnancy (13.2± 2.1 wk).Women completed a 3-d food diary at 16–20weeks of gestation, anddatawere entered into theNutritionData System

for Research (NutritionCoordinatingCenter; University ofMinnesota).A median split for percentage kilocalories from CHO (median: 49%;range: 32–75%) further categorized women into high-CHO comparedwith low-CHO groups (n = 14–16/group). Total GWG was calculatedas the difference in body weight measured at enrollment and at the lastprenatal care visit (time between visits: 180 ± 22 d; did not differ bygroup). Univariate analysis of covariance was used to examine whetherthe association of weight status with GWG differed for women in thehigh-CHO compared with low-CHO groups, after adjustment for totaldaily energy intake.

Results: A significant interaction between weight status and CHOcontent of the diet was found (P < 0.05), such that there was nodifference in GWG for women who were normal weight and consumeda high-CHO diet compared with a low-CHO diet (12.5± 4.5 comparedwith 15.5 ± 9.1 kg), but for women who were obese, those in the low-CHOgroup had lessGWG than those in the high-CHOgroup (7.9± 4.7compared with 13.6 ± 7.1 kg). The interaction did not change afteradjusting for total energy intake.

Conclusions: Results suggest that intake of a relatively low-CHOdiet may help limit GWG among women with obesity. It is possible thatthe difference in GWG by CHO content of the diet was evident onlyfor women with obesity because of differences in glucose metabolismand insulin action compared with women of normal weight. Additionalstudies should establish the feasibility of using a low-CHO diet as anintervention to limit GWG in women who are obese at conception.

Funding SourcesNational Institute of Diabetes and Digestive and Kidney Disease:

K01DK090126 (awarded to PCC), P30DK079626, and P30DK056336;the National Heart Lung and Blood Institute: T32HL105349 (UABPre-Doctoral Training in Obesity-Related Research); and the Na-tional Center for Advancing Translational Sciences: UL1TR001417,at the NIH.

Effects of Inflammation and Ketoisocaproic Acid on GlucoseMetabolism in Muscle Cells (OR09-01)

Olasunkanmi J Adegoke and Gagandeep Mann

York University, United Kingdom

Background: Recent studies have linked elevated concentrationsof branched-chain amino acids (BCAAs) and their metabolites to thepathogenesis of insulin resistance and type 2 diabetes. We previouslyshowed that α-ketoisocaproic acid (KIC), a metabolite of leucine,inhibits insulin-stimulated glucose uptake but only if branched-chainaminotransferase 2, the enzyme that catalyzes the reversible conversionof leucine to KIC, is present. Inflammation is a major component ofinsulin resistance, so the effect of KIC on insulin resistance may bemodified by the inflammatory environment.

Objective: The aim of this study was to determine whether or notKIC’s role in insulin stimulated glucose transport in L6myotubes wouldbe modified by coincubation with homocysteine, a proinflammatoryfactor.

Results: KIC (200 mM) suppressed insulin stimulated glucoseuptake by 25%. There was further suppression (7% and 34%) of insulin-stimulated glucose transport by coincubation with 50 and 500 mMhomocysteine, respectively. Interestingly, coincubation of muscle cellswith KIC and homocysteine increased AKT ser 473 phosphorylation.


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Because the effect of homocysteine was rather modest, we attemptedto produce a more robust inflammatory response by coincubatingthe myotubes with additional proinflammatory factors. KIC with 10ng/ml of tumor necrosis factor-α, 50 mM homocysteine, and 10 ng/mlof interleukin-6, resulted in a further decrease of 43% in insulin-stimulated glucose uptake compared with incubation with KIC alone.

Conclusions: These data suggest that the effect of amino acids andtheir metabolites on insulin action in skeletal muscle are modulatedby the inflammatory environment seen in obesity and insulin-resistantstates. Interventions that modulate BCAA metabolism, especiallywithin the context of inflammation, can have implications on themanagement of insulin resistance and its comorbidities.

Funding SourcesNSERC (Canada).

Enteroids Recapitulate in vivo Citrulline Production in Mice(OR09-02)

Juan Marini, Xiaoying Wang, Yuan Yang, Inka C Didelija, andMahmoud A Mohammad

Baylor College of Medicine, TX

Background: The endogenous production of arginine relies on thesynthesis of citrulline by enteral ornithine transcarbamylase (OTC).Mutations in the gene coding for this enzyme are the most frequentcause of urea cycle disorders. However, there is a lack of correlationbetween in vivo metabolic function and DNA sequence, transcriptabundance or in vitro enzyme activity. Enteroids, a novel ex vivomodel,may be able to replicate the in vivo metabolic phenotype.

Objective: The goal of the present work was to test the hypothesisthat enteroids recapitulate the in vivo citrulline production of wild-type(WT) and spf-ashmice, a hypomorphic model of OTC deficiency.

Methods: Male WT and OTC-deficient mice (spf-ash mutation),6 wk old, were studied. Urea, citrulline, and arginine fluxes weredetermined in vivo, and OTC abundance was measured in liver and guttissue. Intestinal crypts were isolated and cultured to develop enteroids.Ex vivo citrulline production and OTC abundance were determined inthese enteroids.

Results: Liver OTC abundance was lower (P < 0.001) in spf-ashthan in WT mice (0.16 ± 0.01 vs. 1.85 ± 0.18 a.u.), but there wasno difference (P = 0.32) in urea production. In gut tissue, OTCwas barely detectable in the mutant mice; despite this, a lower butsubstantial citrulline production (67 ± 3 vs. 167 ± 8 µmol · kg–1 · h–1,P < 0.001) was determined in the mutant mice. This translated intolower arginine fluxes and plasma concentrations (320± 24 vs. 465± 26µmol · kg–1 · h–1, P < 0.001; 63 ± 9 vs. 132 ± 3 µmol/L, P < 0.001;respectively). The enteroids recapitulated the in vivo findings: a verylow OTC abundance accompanied by a reduced citrulline production(1.07 ± 0.20 vs. 4.64 ± 0.44 nmole/µg DNA, P < 0.001).

Conclusions: In conclusion, enteroids recapitulated in vivo cit-rulline production and offer the opportunity to study the regulationof citrulline production in a highly manipulable system. In addition,similarly to its in vivo role as a marker for gut mass and function,citrulline may be a useful marker for ex vivo metabolic function.

Funding SourcesUSDA, Agricultural Research Service under Cooperative Agree-

ment No. 58-3092-5-001, NIH (R01 GM108940).

Gut Microbiota and Dietary Fiber Mitigate Amino Acid StressResponse in Low-Protein Diets (OR09-03)

Anthony Martin,1 Connie Ha,1 Gustaf Hendrick,1 Donald KLayman,2 and Suzanne Devkota1

1Cedars-Sinai Medical Center, CA; and 2University of Illinois atUrbana-Champaign

Objective: Experimental low-protein diets (LP) (<10% of totalenergy) can create amino acid (AA) deficiencies and induce stressresponses, including increased circulating FGF21 and upregulationof genes associated with AA biosynthesis (Asns and 3pgdh) andendoplasmic reticulum (ER) stress (eIF2a), yet LP vegetarian dietsseldom exhibit overt AA deficiency. Recent studies have demonstratedthat gut microbiota can play a significant role in the nutrition status ofthe host. In ruminant animals, for example, the microbiota is knownto utilize plant biomass to produce essential AAs. We hypothesize thatmicrobiota in monogastric species may also adapt to protect the hostfrom AA deficiency, depending on the quality and quantity of fiber inthe diet. The aim of this study was to test that hypothesis.

Methods: C57BL/6 mice were fed 1 of 5 diets: a protein-sufficientdiet (AIN-93G-based control) or 10% LP diets supplemented with 5%cellulose (5C), 15% cellulose (15C), 5% inulin (5I), or 15% inulin (15I)for 3 wk (n = 10/group). Protein deficiency was assessed via plasmaFGF21, hepatic gene expression of Fgf21, Asns, and 3pgdh, and hepaticp-eIF2a. Fecal microbiota was characterized at baseline and after 3 wkby 16S rRNA sequencing. Statistical significance was determined byone-way ANOVA and Tukey or Dunnett post hoc tests for multiplecomparisons.

Results: Cellulose-supplemented LP diets attenuated ER stresscompared with inulin-based diets, evidenced by significantly lowerFGF21, Fgf21, and p-eIF2a. The 15C diet in particular showed nodifference compared with the protein-sufficient control diet. Further-more, expression of Asns and 3pgdh was not significantly upregulatedon the 5C and 15C diets, suggesting protein sufficiency. Both cellulosediets promoted blooming of Lactococcus and Blautia, as well asRuminococcus, which is a known cellulose degrader in ruminants.Changes in the abundance of these genera across treatment groupssignificantly and inversely correlated with plasma FGF21.

Conclusion: These results demonstrate that increased cellulose-based fiber consumption can attenuate protein deficiency through apotentially microbially linked mechanism. Most cellulose is nonfer-mentable by human enteric bacteria; therefore, the mechanism ofprotection may not be because of direct carbon utilization of cellulosefor de novo AA biosynthesis, but rather a supportive role for growth ofbeneficial species with potential to impact protein or AA status.

Assessment of Encapsulated Leucine as a Potential AnabolicAgent to Enhance Lean Growth in Neonates (OR09-04)

Agus Suryawan,1 Marko Rudar,1 Jane Naberhuis,1 HanhNguyen,1 Marta L Fiorotto,1 Zhefeng Li,2 and Teresa A Davis1

1USDA/ARS Children’s Nutrition Research Center, Baylor Collegeof Medicine, TX; and 2Hangzhou King Techina Feed Co., Ltd, China

Objective: The rise in amino acids and insulin after a meal stimu-lates protein synthesis in skeletal muscle of neonates, and the responseto amino acids is largely driven by leucine. Leucine supplementation


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enhances protein synthesis, but the effect wanes as the concentrationof leucine falls. The objective of this study was to determine if leucinesupplementation can prolong the anabolic stimulus of feeding if givenin a slowly released form so that the leucine peak occurs after the insulinsurge.

Methods: Piglets (5 d old, n= 3–4) were randomly assigned to 1 of 3dietary treatments: 1) control (C) protein-restricted diet [10.5 g protein· kg body weight (BW)–1 · d–1; 2) C diet + 1.19 g · kg BW–1 · d–1 free-form leucine (Leu); and 3) C diet + 2.39 g · kg BW–1 · d–1 encapsulatedleucine (50% leucine+ 50% encapsulated materials) (ELeu). Diets werefed every 4 h for 24 h, then blood samples were collected at intervalsafter the last meal. A flooding dose of L-[4-3H]Phe was infused at 210min, and 30 min later longissimus dorsi muscle samples were collectedfor determination of fractional protein synthesis rate and mechanistictarget of rapamycin complex 1–regulated translation initiation signalingactivation.

Results: Plasma leucine concentrations were higher in ELeu than inLeu and C at 210 min (means ± SEMs: 419 ± 38, 292 ± 38, and 87 ±12 μM, respectively; P < 0.05). Plasma isoleucine (Ile) and valine (Val)concentrations were lower in Eleu and Leu than in C (Ile = 54 ± 4,42 ± 5, and 184 ± 8 μM; Val = 56 ± 9, 76 ± 9, and 381 ± 7 μM,respectively; P< 0.05). Circulating concentrations of most other aminoacids did not differ between groups. Phosphorylation of S6K1 and4EBP1 and formation of eIF4G-eIF4E complex in muscle were greaterin ELeu than in Leu and C (P< 0.05). Fractional protein synthesis ratesdid not differ between groups.

Conclusions: Supplementation of a low-protein diet with ELeudelayed the release of leucine, resulting in prolonged activation ofkey mTORC1 signaling proteins involved in translation initiation inneonatal muscle. The limited availability of Ile and Val as substratesfor protein synthesis likely contributed to the lack of effectiveness ofELeu in stimulating muscle protein synthesis. Further study is neededto determine whether the use of ELeu, while maintaining substrateavailability, could be an effective strategy to enhance muscle proteinsynthesis and lean growth in the neonate.

Funding SourcesNIH HD085573, USDA CRIS 6250-51,000-055, NIH HD072891,

USDA NIFA 2013-67,015-20,438.

Protein and Carbohydrate Supplementation Modulate SkeletalMuscle MicroRNA following Aerobic Exercise (OR09-05)

Lee M Margolis,1 Claire Berryman,1 Nancy Murphy,1 Christo-pher Carrigan,1 Andrew J Young,1 John Carbone,2 and Stefan MPasiakos1

1US Army Research Institute of Environmental Medicine; and2Eastern Michigan University

Background: Skeletal muscle microRNAs, which inhibit musclehypertrophy through negative regulation of the mechanistic target ofrapamycin complex 1 pathway, are diminished and inversely associatedwith muscle protein synthesis after aerobic exercise when protein andcarbohydrate are consumed together during exercise. Whether thereare independent effects of recovery protein and carbohydrate nutritionon microRNA and their potential to modulate anabolic and proteolyticregulators of muscle mass after exercise are largely unknown.

Objective: The aim of this study was to examine effects of recoverywhey protein (WP) ingestion, with or without carbohydrate, on skeletalmuscle microRNA after exercise.

Methods:Vastus lateralis samples were obtained in 17 healthymalesbefore (BASE) and after 80 min of steady-state treadmill exercise(maximal oxygen uptake: 1.7 ± 0.3 L/min), followed by a 2-miletime trial (17.9 ± 3.5 min; POST), and 3 h into recovery (REC).Participants were randomized to consume either a carbohydrate (CHO;145 g; n = 9) or a non-nutritive placebo (PLA; n = 8) drink duringexercise. All participants consumed a bolus drink of WP (25 g)immediately after exercise. Anabolic and proteolytic signaling wereassessed using Western blotting, and microRNA and mRNA weredetermined using quantitative reverse transcriptase-polymerase chainreaction. Bioinformatics analysis for predictive markers of microRNAwas completed using microT-CDS software.

Results: Independent of carbohydrate, the concentrations of theinhibitors ofmuscle anabolismmiR-1–3p,miR-133b, andmiR-206werelower (P < 0.05) at POST and REC than at BASE. p-rpS6Ser240/244, amolecular regulator of anabolism, was higher (P < 0.05) at POST andREC than at BASE, regardless of carbohydrate. Independent of time,miR-222–3p and miR-324–3p concentrations were higher (P < 0.05)in the CHO group than in the PLA group, and miR-486–5p was higher(P < 0.05) at REC in the CHO group than in the PLA group. PTEN,an inhibitor of AKT, was identified as a common target of miR-222–3p and miR-486–5p. PTEN was lower (P < 0.05) at REC in the CHOgroup than in the PLA group, and downstream markers of proteolysis,p-FOXO1Thr24 and MuRF1, were also lower (P < 0.05) at POST in theCHO group than in the PLA group.

Conclusion: These data suggest that carbohydrate ingested duringexercise supports muscle mass accretion potentially through microR-NAs that inhibit skeletal muscle proteolysis, with little effect onmicroRNAs that regulate anabolism.

Funding SourcesThis project was supported by the US Army Medical Research and

Materiel Command.

Decrease in Endogenous Substrate Flux Is the Principal Mecha-nism Underlying the Reduction in Plasma Branched-Chain AminoAcids following Bariatric Surgery (OR09-06)

Hong Chang Tan,1 Jean Hsu,2 SIdney Yu,1 Jean-Paul L Kovalik,3Chin Meng Khoo,4 Oi Fah Lai,1 and Farook Jahoor2

1Singapore General Hospital; 2Baylor College of Medicine, TX;3Duke-NUS Graduate Medical School, Singapore; and 4National Uni-versity Health System, Singapore

Background: Branched-chain amino acids (BCAAs) and branched-chain ketoacids (BCKAs) are elevated in morbid obesity. Thesemetabolites decrease significantly after bariatric surgery, but it is unclearwhether a reduction endogenous substrate flux (Ra) or increase inclearance is responsible.

Objectives: Our study examined BCAA kinetics in morbidlyobese subjects undergoing bariatric surgery using stable-isotope tracermethods. We hypothesized that the reduction in plasma BCAAs andBCKAs after surgery will be accompanied by significant changes insubstrate Ra and oxidation rates.


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Methods: A total of 10 morbidly obese subjects scheduled forbariatric surgery were studied at baseline and 6 mo after surgery.Leucine Ra, oxidative, and nonoxidative disposal were measured using[U-13C6]leucine, and body composition was measured using dual-energy X-ray absorptiometry. Leucine oxidative capacity was expressedas the percentage of leucine Ra oxidized. Kinetic parameters areexpressed as µmol · kg lean body mass–1 · h–1.

Results: All subjects completed their baseline studies, but 2 did notreturn for their follow-up studies. At 6 mo following surgery, subjectshad experienced a significant decrease in total weight (from 110.5± 5.7to 89.2 ± 5 kg), which was mainly attributed to fat mass loss (68% oftotal weight loss). Three were significant decreases in Leucine Ra (from197.7 ± 6.47 to 148.3 ± 5.8), oxidative (from 21.5 ± 0.98 to 14 ± 2.2),and nonoxidative disposal (from 187.5 ± 6.6 to 145.3 ± 5.5), despitea significant increase in the dietary protein intake from 17.5% ± 2.7%to 23.2% ± 2.2% of total calorie intake. Plasma BCKAs also decreased(from60.35± 4.19 to 46.71± 4.16µmol/L). Importantly, the percentageleucine flux oxidizedwas also reduced after surgery (from10.4%± 0.5%to 8.8% ± 0.5%).

Conclusion: Reduction in endogenous BCAA flux, rather thanimprovement in BCAA clearance, is responsible for postoperativechanges in plasma BCAAs and BCKAs.

Funding SourcesSingHealth and Duke-NUS Collaborative Grant (SDC/020/2015).

FIGURE 1 Plasma branched-chain ketoacids before and afterbariatric surgery.

FIGURE 2 Leucine kinetics before and after bariatric surgery.

Consuming Higher-Protein Breakfasts, Varying in Type, onPostprandial Appetitive, Hormonal, and Neural Responses in Com-bination with Daily Food Intake in Healthy Adults (OR09-07)

Jess A Gwin and Heather Leidy

Purdue University, IN

Objectives:Nutrition messaging surrounding the benefits of break-fast consumption compared with skipping breakfast on appetite controland satiety remain conflicting, particularly owing to the inconsistentquality and type of breakfasts provided. Although breakfasts containinghigher-protein (HP) foods elicit improvements in these outcomes, itis unclear as to whether HP beverages elicit similar responses. Thus,the purpose of this study was to examine the effects of consuming HPbreakfasts, varying in breakfast type, on daily appetite, satiety, and foodintake vs. skipping breakfast in healthy adults.

Methods: A cohort of 13 adults (age: 23.5 ± 0.9 y; body mass index:23.6± 0.6 kg/m2) completed the following acute randomized crossovertrial. The participants consumed an HP breakfast (350 kcal; 30 g ofprotein, 35 g of carbohydrate, 10 g of fat) in either FOODorBEVERAGEform or skipped breakfast (SKIP) for 7 d per pattern. On day 7 ofeach pattern, a tightly controlled 8-h clinical testing day was completed,consisting of repeated assessments of appetite and satiety sensations incombination with circulating hormones (plasma ghrelin, PYY, orexin-A). Food-stimulated functional MRI was also performed at 240 minpostbreakfast or prelunch. Ad libitum food intake and food choice wereassessed throughout the day.

Results: Postprandial hunger, desire to eat, and prospective foodconsumption decreased, whereas fullness increased, following theconsumption of bothHP breakfasts compared with SKIP (all, P< 0.05),with no differences occurring between the FOOD and BEVERAGEbreakfasts. Both HP breakfasts reduced postprandial ghrelin andincreased PYY concentrations compared with SKIP (all, P < 0.05),with no differences between the breakfast types. Postprandial orexin-Aconcentrations were no different between treatments. Functional MRIresults are pending. Evening snacking consumption onhigh fat and highcarbohydrate foods was reduced following the HP breakfasts (FOOD,650 ± 123 kcal, P < 0.05; BEVERAGE, 781 ± 185kcal, P < 0.05)compared with SKIP (938 ± 180kcal); no differences were detectedbetween FOOD and BEVERAGE.

Conclusions: The consumption of breakfasts containing higher-protein foods or beverages similarly improve select markers of appetite,satiety, and snacking behavior compared with skipping breakfast inhealthy adults.

Funding SourcesEgg Nutrition Center.

Dietary Intakes and Circulating Levels of Branched-ChainAmino Acids in Relation to Long-Term Incident Type 2 DiabetesRisk in a Prospective Cohort of Women (OR09-08)

Deirdre Tobias,1,2 Patrick Lawler,3 Aruna D Pradhan,1 Paulo HHarada,1 Olga Demler,1 Paul Ridker,1 Susan Cheng,1 Frank BHu,4 JoAnn Manson,1 and Samia Mora1

1Brigham and Women’s Hospital, MA; 2Harvard Medical School,MA; 3Peter Munk Cardiac Centre, University Health Network, andHeart and Stroke Richard Lewar Centre of Excellence in Cardiovascular


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Research, University of Toronto, Canada; and 4Harvard TH ChanSchool of Public Health, MA

Objectives: The role of dietary intakes of branched-chain aminoacids (BCAAs: isoleucine, leucine, valine) with type 2 diabetes (T2D)risk is unclear. However, circulating BCAAsmetabolites are consistentlyassociated with an elevated T2D risk, collectively implicating impairedBCAA metabolism as a relevant predictor of T2D risk. We aimed toevaluate the individual and joint associations for dietary and plasmametabolite concentrations of BCAAs with incident T2D.

Methods: Participants enrolled in the longitudinal US-basedWomen’s Health Study cohort were eligible for analysis if they were freeof T2D, cancer, or cardiovascular disease at the baseline blood collection(n = 25,972, mean baseline age = 54.6 y). We estimated BCAAintakes from the baseline food frequency questionnaire. Plasma BCAAmetabolites were measured via proton nuclear magnetic resonancespectroscopy, ln-transformed, and standardized for analysis. We usedmultivariable Cox proportional regression models to estimate HRs and95% CIs for quintiles of diet and plasma BCAAs with incident T2Drisk, adjusting for age, total calories, body mass index, physical activity,family history, and other established risk factors. We also assessed jointexposure to below or above medians of diet and plasma concentrations,with lower diet or lower plasma as reference.

Results: We confirmed 2364 incident T2D cases over a mean 19-yfollow-up. Total BCAA plasma concentrations, but not dietary intakes,were positively associated with incident T2D [Q5 vs. Q1: metaboliteHR: 3.0 (95% CI: 2.6, 3.5); diet HR: 1.0 (95% CI: 0.8, 1.3)]. Modelingthe joint association indicated that higher circulating BCAAs wereassociated with T2D regardless of dietary intake. In addition, higherdietary BCAAs were not associated with T2D if plasma concentra-tions were not elevated. Similar results were achieved for individualBCAAs.

Conclusion: Higher plasma BCAAs, but not dietary intakes, wereassociated with an elevated incident T2D risk in women. These findingsare consistent with prior evidence that it is the underlying impairedBCAA metabolism, rather than diet per se, that is the relevant markerof T2D risk.

Funding SourcesFunding was from NIH grants CA047988, HL043851, HL080467,

HL099355, UM1 CA182913, K01 DK103720, and DK112940, and agrant from the American Heart Association (0670007N).

CoconutOil Goes Nuclear: The Impact of Dietary Lauric Acid onthe Production of a Ligand for LRH-1 (OR17-01)

Kevin C Klatt, Olga Malysheva, Mark Roberson, and MarieCaudill

Cornell University, NY

Background: Liver receptor homolog-1 (LRH-1) is a nuclear recep-tor with notable roles in regulating sterol metabolism. To date, LRH-1’s endogenous ligand remains unknown; however, recent analyseshave suggested that an “uncommon” phosphatidylcholine species,dilauroylphosphatidylcholine (DLPC), binds and activates LRH-1 bothin vivo and in vitro. Controversy exists about the relevance of this ligand,as it has yet to be identified endogenously across model systems.

Objective: The aim of this study was to test the hypothesis thatprovision of lauric acid (C12) to model systems would result in DLPCproduction.

Methods: We utilized HepG2 cells, C57BL/6J mice, and humanparticipants to test our hypothesis. Cells were cultured in fetal bovineserum-containing media supplemented with C12-bound bovine serumalbumin (BSA), oleic acid-BSA (18:1), or fatty acid free BSA for 4 h, withand without an LRH-1 antagonist. C57BL/6J mice were fed diets withoils containing C12 (olive oil+C12) or coconut oil (∼49%C12) in bothacute and chronic conditions. A pilot trial was undertaken to assess theimpact of consuming a breakfast shake containing coconut oil onDLPCin postprandial serum. DLPC in cell lysates, animal tissue, and humanserum was assessed using LC/MS.

Results: In all 3 model systems, provision of lauric acid in themedia or diet resulted in the production of DLPC. Chronic feedingresulted in DLPC production in murine intestine, liver, pancreas,muscle, and ovary; consumption of C12 by human participants resultedin DLPC in postprandial serum; this effect is being further tested inan ongoing randomized, controlled, crossover feeding study. In HepG2cells, media supplemented with C12 upregulated hepatic LRH-1 targetgene expression (CYP7A1, 6.6 ± 2.87; CYP8B1, 2.22 ± 0.66; P < 0.05),an effect that was ablated by cotreatment with an LRH-1 antagonist.Consistent with the previously reported beneficial effect of DLPCsupplementation on glucose tolerance in high-fat diet–fed animals,coconut oil–fed mice maintained fasting blood glucose similar to thatof chow-fedmice (coconut oil: 191.57± 10.35; olive oil: 253.33± 13.58;chow: 182.83 ± 9.7; P < 0.001) despite similar weight gain.

Conclusion: Provision of lauric acid in media or diet results in theproduction of the ligand for the orphaned nuclear receptor LRH-1, andsuggests the potential for diverse health effects of this saturated fattyacid beyond its influence on blood cholesterol concentrations.

Funding SourcesNIH Training Grant T32HD087137.

Activation of a Hepatic cAMP-Responsive Element-BindingProtein H-Apolipoprotein A5 Axis Increases Peripheral VLDL-Triglyceride Clearance in Response to Dietary Protein Restriction(OR17-02)

Jose Humberto Trevino-Villarreal,1 Justin Reynolds,1 AlexBartelt,1 Kent Langston,1 Michael MacArthur,1 Sarah Mitchell,1Kaspar Trocha,1 Joerg Heeren,2 Keith Ozaki,3 Luigi Fontana,4and James Mitchell1

1Harvard School of Public Health, MA; 2University of Hamburg,Germany; 3Harvard Medical School, MA; and 4Washington Universityin St. Louis

Objectives: Hypertriglyceridemia is an independent risk factorfor cardiovascular disease. Dietary protein restriction (PR) withoutenforced calorie restriction reduces triglycerides (TGs) in preclinicalmodels, but the underlying molecular mechanisms and the clinicaltranslational potential remain unknown. Here, we sought to 1) identifythe molecular mechanisms underpinning reduction of VLDL-TGsduring PR and 2) determine the potential clinical translation in humans.

Methods: Lean or diet-induced obese mice—B6D2F1 or LDL-receptor knockout (LDLR-KO) on a C57BL/6 background—were givenad libitum access to a semipurified complete diet (18% protein, 22% fat,


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60% carbohydrates) or a PR diet (where protein was replaced with anisocaloric amount of sucrose) for 7 d. TGs were measured in plasma inthe fasted state. Hepatic cAMP-responsive element-binding protein H(CREBH) activation and apolipoprotein A5 (ApoA5) expression weremonitored by immunoblotting and quantitative reverse transcriptase-polymerase chain reaction. Plasma mouse and human ApoA5 weredetermined by ELISA. The functional roles of ApoA5 and CREBHwereassessed using global knockout models. The rate of VLDL-TG disposalwas determined by injecting purified, radiolabeled VLDL particlesprepared from mice on complete or PR diets. In humans, PR consistedof 4–6 wk on a plant-based diet with 8–9% of total calories fromprotein.

Results: PR significantly lowered TGs in lean mice, diet-inducedobese mice, and LDLR-KO mice fed with a PF-high-fat diet. Mech-anistically, this occurred in part via induction of the TG-loweringapolipoprotein, ApoA5. Increased hepatic and VLDL ApoA5 proteinexpression resulted in more rapid VLDL-TG clearance. Activationof the hepatic transcription factor CREBH occurred in response to

FIGURE 1 PR reduces circulating VLDL-TGs in lean mice, and in high-fat diet mice on a wild type (B6D2F1), or a LDLR-KO geneticbackground. B6D2F1 mice or LDLR-KO on a C57BL/J background were fed the indicated complete or protein free (PF) diet for 1 wk priorto analysis. (A) Plasma TG concentrations in mice fed on a complete (18% protein content) or isocaloric PF diet (protein replaced withisocaloric sucrose), provided on an ad libitum basis (AL) or restricted daily (CR) by 50% (n = 5 per group; one-way ANOVA with Dunnett’spost-test compared to the complete AL group) (B) Representative chromatogram of comparative serum lipidomic analysis. Arrow points tothe peaks magnified in the inset, corresponding to serum TGs. (C) Plasma TG concentrations from mice fed ad libitum on the indicatedcomplete (comp.) or PF diet and collected in the fed or fasted state (n=5 per group; Student’s T test between diets within fed or fastedstate). (D) TG measurements of plasma FPLC fractions (derived from a pool of 5 mice per group). Fractions containing differentlipoproteins are indicated above (VLDL, LDL, HDL). (E) Plasma TG concentration in fed and fasted states (n = 8 per group; Student’s T testwithin fed or fasted state). (F) Fasted plasma TG concentrations. Data expressed as mean ±SD; * P < 0.05, * *P < 0.01, * * *P < 0.001.

PR, and was required for the increase in Apoa5 mRNA expressionduring PR. Genetic ApoA5 and CREBH knockout models were bothhypertriglyceridemic on a complete diet and failed to fully lowerTG upon PR. In humans, PR reduced circulating TG concentrationsconcomitant with an increase in ApoA5 protein.

Conclusions: PR reduced circulating TG by altering the apolipopro-tein composition of VLDLparticles, promoting the peripheral clearanceof TG. This required activation of the dietary protein responsivetranscription factor, CREBH, and its target, ApoA5. Conservation ofPR-mediated TG lowering in humans suggests reduced dietary proteinintake as a potential intervention for hypertriglyceridemia.

Funding SourcesFunded by American Heart Association Grant-in-Aid 16GRNT

27090006 NIH (RO1DK090629, R01AG036712) and Charoen Pokp-hand Group Bakewell Foundation, the Longer Life Foundation (anRGA/Washington University Partnership), the National Center forResearch Resources (UL1 RR024992), and a Deutsche Forschungsge-meinschaft Research Fellowship (BA 4925/1-1).


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FIGURE 2 PR alters VLDL-TG clearance via changes in hepatic apolipoprotein expression. B6D2F1 mice were fed the indicated diet for1 wk prior to analysis of hepatic parameters. (A) Steady-state hepatic mRNA levels of Apoa2, Apoc2, Apoc3, and Apoa5 (n=10 per group;Student’s T test). (B) Immunoblot of hepatic ApoA5 protein expression. Below, quantitation normalized to tubulin (n = 5 per group, usinga Student’s T test). (C) Representative composed images obtained using confocal microscopy showing induction of ApoA5 (red)expression and co-localization with the ER marker KDEL (green) in liver; DNA from nuclei in blue. Images are superimposed to adifferential interference contrasted image of the respective area (scale bar = 20 μm). (D) Ratio of ApoA5 to ApoB-100 expression in4hr-fasted plasma as a measure of ApoA5 concentration per VLDL particles (n = 7-10 per group; Student’s T test). (E) Correlation analysisbetween plasma TGs and circulating ApoA5. Each dot represents an individual mouse; r, Pearson’s coefficient. (F) Fasted plasma TG levelsin whole body ApoA5-KO or control FVB wildtype mice on the indicated diet for one week; one-way ANOVA with Sidak’s multiplecomparisons test as indicated. Data expressed as mean ±SD; *P < 0.05, **P < 0.01, ***P < 0.001; ns, not significant.

FIGURE 3 PR alters VLDL-TG consumption via changes in hepatic apolipoprotein expression. (A) Silver stain of SDS polyacry lamide gelfrom serum fractions from mice fed the indicated complete (C) or protein free (PF) diet for one week and then separated by densitycentrifugation. Serum, load: VLDL, top fraction: F2 and F3, fractions 2 and 3. ApoB100 protein is characteristic of VLDL particles Sampleswere nomalized within fraction groups by protein content resolved using denatured SDS-polyacrylamide gel electrophoresis, andvisualized using sliver staining. (B) Plasma 3H levels over time after injection of [3H]palmitate-labeled VLDL particles purified fromcomplete or protein free (PF) diet fed mice as indicated and injected into fasted mice on a complete diet and expressed as a percentageof total radioactivity injected. Differences in clearance rate were analyzed using paired t-test between groups at each different timeintervals (n = 4 per group).


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FIGURE 4 CREBH regulates ApoA5 expression upon PR. (A-D) B6D2F1 mice were fed the indicated complete or protein free (PF) diet for1 wk prior to analysis. (A) Immunoblot of CREBH full-length (FL) and cleaved forms (CF) in whole liver extracts. The first two lanes containliver extracts from CREBH-KO mice to control for antibody specificity. Below, quantitation of cleaved CREBH normalized to full-length(n = 5 per group; Student’s T test). (B) Immunoblot of cleaved CREBH in hepatic nuclear extracts. Below, quantitation normalized to laminA/B (n = 5 per group; Student’s T test). (C) Representative confocal microscopic images of livers stained for the ER-binding sequenceKDEL (green), the amino-terminus CREBH fragment (red), and nuclei with 4’ 6-diamidino-2-phenylindole (DAPI, blue). Magnification: 63X;scale bars =30 μm (D) Confocal microscopy image of HepG-2 cells transfected with a plasmidencoding an HA-tagged cleaved fragmentof human CREBH (hCREBH(N)) or empty vector (mock). Nuclear co-localization was confirmed by using an Alexa-488 conjugated anti-HAantibody and DAPI for nuclear visualization. White scale bar = 20μm (E) Steady-state mRNA levels of ApoA5 in human HepG-2 cellstransfected with a plasmid encoding an HA-tagged cleaved fragment of human CREBH (hCREBH (N)) or empty vector (mock) as control.Data represents triplicates from one experiment; Student’s T test. The experiment was repeated two times with similar results. (F-I)CREBH-KO or WT control littermates were fed the indicated diet for 1 wk prior to analysis (n = 5-8 per group; data analyzed by one-wayANOVA with Sidak post-test). (F) Steady-state hepatic mRNA levels of Apoa5. (G) Immunoblot of hepatic ApoA5. Below, quantitationnormalized to tubulin. (H) Expression of ApoA5 in VLDL particles. Estimation of ApoA5 per particle was obtained by normalizing plasmaApoA5 against plasma ApoB-100. (I) Fasted plasma TG concentrations in CREBH-KO and control WT littermates on the indicated diets(n = 8 in this experiment). The Experiment was repeated 4 times with similar outcomes. Data expressed as mean ±SD; *P < 0.05,**P < 0.01, ***P < 0.001.


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FIGURE 5 PR increases ApoA5 expression and reduces TG levels in humans. (A-B) Immunoblot analyses of CREBH cleavage(A) and ApoA5 protein expression in whole liver extracts of mice subject to different degrees of PR as indicated for one week. Below,quantitation (n = 4/group; one-way ANOVA with Dunnett’s post-test compared to the 18%-complete protein group). (C) Serum TGs frommice subject to different degrees of PR as indicated; one-way ANOVA with Dunnett’s post-test compared to the 18%-complete proteingroup. (D) VLDL-ApoA5 expression assessed by determining the ApoA5/ApoB ratio and (E) circulating TGs in plasma from humansubjects (n = 19) before and after 4-6 weeks on an 8-9% protein diet. Statistical analysis was performed using a non-parametric two-tailedpaired t test and corrected using a Wilcoxon post-test. (F) Correlation between ApoA5 and plasma TGs from patients before and after PR;non-parametric Spearman’s correction (r = —0.3643).

Resistance to High-Fat Feeding–Induced Decline in ExerciseCapacity in Liver Fatty Acid–Binding Protein Null Mice (OR17-03)

Heli Xu, Angela Gajda, Yin Xiu Zhou, Anam Fatima, and JudithStorch

Rutgers University, NJ

Objective: The aim of this study was to study the metabolicalterations in skeletal muscle of liver fatty acid–binding protein null(LFABP–/–) mice underlying their resistance to high-fat feeding–induced decline in exercise capacity.

Methods:Resting skeletal muscles of LFABP–/– mice were examinedfor glycogen and intramuscular triglyceride (IMTG) concentrations,mitochondrial enzyme activities, fatty acid oxidation capacity, andtranscriptional factors involved in lipid metabolism. The respiratoryexchange ratio (RER) was also measured in LFABP–/– mice during alow-intensity treadmill test. Postexercise glucose and free fatty acids(FFAs) were determined in the plasma of LFABP–/– mice, and theirpostexercise skeletal muscles were examined for IMTG and glycogenstores.

Results: Compared with wild-type (WT) mice, resting skeletalmuscles of LFABP–/– mice had higher concentrations of glycogen

and IMTG, as well as an increased fatty acid oxidation rate andgreater mitochondrial enzyme activities, suggesting a higher substrateavailability and substrate utilization capacity. Interestingly, based on thedynamic changes in the RER during exercise, LFABP–/– mice utilizedmore carbohydrate at the beginning of the exercise, then switchedto using lipids preferentially in the later stage. The LFABP–/– miceconsistently exhibited a greater exercise-dependent decrease in muscleglycogen stores and elevated circulating FFAs after exercise.

Conclusions: The improved exercise capacity in high fat–fedLFABP–/– mice is related to greater basal concentrations of energysubstrate and mitochondrial function in skeletal muscle, as well asmore efficientmuscle glycogen and fatty acid utilization during exercise.Since LFABP is not expressed in the skeletal muscle, but rather inliver and intestine, these muscle metabolic changes in LFABP–/– micemay, in part, be because of liver- and/or intestine-derived mediatorscontributing to the prevention of high-fat feeding–induced impairmentin the skeletal muscle.

Funding SourcesThis work was supported by NIH Grant DK-38,389 from NIDDK,

and by funds from the New Jersey Agricultural Experiment Station(to JS).


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FIGURE 1 Exercise capacity.

FIGURE 2 Increased basal concentrations of IMTG and glycogen.

FIGURE 3 Increased mitochondrial enzyme activities.


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FIGURE 4 Average RER during exercise.

FIGURE 5 Exercise-dependent decrease in muscle glycogen and postexercise plasma FFA.

Comparison of the Postprandial Metabolic Fate of 13C-LabeledStearic and Oleic Acids in Postmenopausal Women (OR17-04)

Jose Rodriguez-Morato,1,2 Jean Galluccio,2 Gregory GDolnikowski,2 Alice H Lichtenstein,2 and Nirupa R Matthan2

1Universitat Pompeu Fabra, Spain; and 2JeanMayerUSDAHNRCA,MA

Objective:Not all dietary saturated fatty acids behave similarly. Sat-urated fatty acids (FAs) 12:0, 14:0, and 16:0 have hypercholesterolemiceffects, whereas stearic acid (18:0) appears to have a neutral effect,similar to oleic acid (18:1). The mechanism is frequently attributedto the in vivo conversion of 18:0 to 18:1. This study was designed tocompare the metabolic fate of 13C18:0 and 13C18:1 in the fed state afterhabituation to diets enriched in the corresponding fatty acid.

Methods: Hypercholesterolemic postmenopausal women (n = 6;age: ≥50 y; body mass index: 25.6 ± 3.0 kg/m2; LDL cholesterol:≥110 mg/dL) consumed both 18:0- and 18:1-enriched diets for 5 wkeach, according to a randomized, controlled crossover trial. The dietsprovided 55% of total energy from carbohydrate, 15% from protein,and 30% from fat, with half of the fat contributed by the 18:0 or 18:1.On day 1 of week 5, following a 12-h fast, participants receive theirusual diet divided into 13 hourly meals beginning at 0800. A purified

tracer (1.0 mg/kg body wt) of [13C]18:0 or [13C]18:1 was incorpo-rated into the meal at 1300. Serial blood and breath samples werecollected throughout day 1, and fasting blood and breath samples werecollected on the mornings of days 2 and 3. The plasma FA profile wasassessed by GC, isotope enrichment by Q-TOF, and FA oxidation rate(expired 13CO2) by isotopic ratio mass spectrometry.

Results: Plasma enrichment curves followed a similar pattern, butthe area under the curve (AUC) of [13C]18:0 was higher than [13C]18:1(9310 vs. 5318 ng · h–1 · mL–1; P < 0.05). Interestingly, [13C]18:0was first converted to [13C]16:0 and desaturated to [13C]16:1, thenelongated to [13C]18:1, the latter being the major metabolite (9%). NoFA metabolites of [13C]18:1 were detected. A higher proportion of 18:0and its metabolites (29%) was incorporated into the plasma triglyceridefraction than 18:1 (8%). Breath 13CO2 kinetic curves indicated that[13C]18:0 had a 34% lower cumulative oxidation rate than [13C]18:1.

Conclusion: Our results indicate that the neutrality of 18:0 onplasma cholesterol concentrations is not attributable to a singlefactor. The higher plasma [13C]18:0 AUC reflects lower oxidationrates, multistage conversion to 18:1, and preferential incorporation,along with its metabolites, into plasma triglycerides. These dataprovide new insights into the differential metabolism of 18:0 and18:1.


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Funding SourcesFunded by HNRCA pilot funds USDA-NIFA-AFRI-003397 and

USDA 1950-51,000-072-02S. JR-M acknowledges funding from theEuropean Union’s Horizon 2020 research and innovation programmeunder the Marie Skłodowska-Curie grant agreement No 712,949(TECNIOspring PLUS) and from ACCIO.

Habituation to Diets Enriched in Palmitate and Stearate Rel-ative to Oleate Differentially Affect Cardiovascular Disease RiskFactors and Markers of Inflammation in Postmenopausal Women(OR17-05)

Nirupa R Matthan,1 Huicui Meng,2 Dayong Wu,2 Lijun Li,2 andAlice H Lichtenstein2

1Tufts University, MA; and 2Jean Mayer USDA Human NutritionResearch Center on Aging at Tufts University, MA

Objective: Dietary fat quality has a significant impact on cardio-vascular disease (CVD) risk, yet the relative effects of stearate (18:0), alonger chain saturated fatty acid (SFA) relative to palmitate (16:0), andto its metabolic product oleate (18:1), a monounsaturated FA, remainunclear. We investigated the effect of consuming diets enriched in 16:0,18:0, or 18:1 on CVD risk factors, indicators of systemic inflammation,and T-cell–mediated function.

Methods: A group of 20 mildly hypercholesterolemic (LDLcholesterol >100 mg/dL) postmenopausal women (age: 50–85 y;body mass index: 25–35 kg/m2) consumed each of 3 diets for 35d according to a randomized, controlled crossover trial. The dietsprovided 55% of the total energy from carbohydrate, 15% fromprotein, and 30% from fat, with half of the fat contributed by 16:0,18:0, or 18:1. The plasma lipid profile and glucose, insulin, andinflammatory and coagulation marker concentrations were measuredusing standard methodology. Lymphocyte proliferation was measuredby [3H]thymidine incorporation following stimulation with anti-CD3/CD28 and T-cell mitogen Concanavalin A by use of a whole-blood assay, and prostaglandin E2 and cytokine production in thepresence or absence of lipopolysaccharides was measured using aradioimmunoassay.

Results: Consumption of the 18:1- and 18:0-enriched diets resultedin significantly lower fasting total cholesterol (TC; –10%, –9%,respectively), LDL cholesterol (–15%, –11%), and HDL cholesterol(–4%, –12%) concentrations (all P < 0.0001) relative to the 16:0-enriched diet. These differences resulted in TC-to-HDL cholesteroland LDL cholesterol-to-HDL cholesterol ratios that were lowest afterconsumption of the 18:1, intermediate after the 18:0 and highest afterthe 16:0-enriched diet. T-cell proliferation in response to antibodiesagainst CD3 (T cell receptor) and CD28 (T cell coreceptor) wassignificantly higher after participants consumed the diet enrichedin 16:0 relative to the diets enriched in 18:0 and 18:1 (54%, 82%,respectively, P = 0.02). There was no significant effect of the dietson plasma triglyceride, glucose, or insulin concentrations, markers ofcoagulation and systemic inflammation.

Conclusion:These data suggest that consumption of a diet enrichedin 18:0 had a similar effect on CVD risk indicators and T-cell–mediatedfunction as a diet enriched in 18:1, and was distinct from those of adiet enriched in 16:0. These findings have implications for dietary SFArecommendations.

Funding SourcesFunded by HNRCA pilot funds USDA-NIFA-AFRI-003397 and

USDA 1950-51,000-072-02S.

Comparing the Consistency of the Serum Triglyceride Responseto High-Dose Supplementation with Either DHA or EPA betweenMen and Women at Cardiovascular Risk: The ComparED Study(OR17-06)

Janie Allaire, Cécile Vors, Johanne Marin, Amélie Charest,Patrick Couture, André Tchernof, and Benoît Lamarche

Laval University, Canada

Objectives: The objectives of this study were 1) to establish theproportion of individuals whose serum triglyceride (TG) concentra-tions are lowered (responders) after high-dose supplementation witheither docosahexaenoic (DHA) or eicosapentaenoic acid (EPA), and 2)to identify factors that predict the TG response toDHA and EPA inmenand women at risk for cardiovascular diseases.

Methods: In a randomized, double-blind, controlled crossoverstudy, 48 men and 106 women with abdominal obesity and subclinicalinflammation were randomized to different sequences of 3 phases:2.7 g/d of DHA, 2.7 g/d of EPA, and 3 g/d of corn oil (0 g ofDHA + EPA). The 10-wk treatment phases were separated by 9-wkwashouts. Blood samples were collected after a 12-h overnight fast atscreening, and at the beginning and end of each phase. Intraindividualvariation in TG, calculated as the standard deviation from the meanof 4 samples taken at screening and before each phase, was equal to±0.25 mmol/L. Responders to DHA and EPA were identified as thosehaving a reduction in TG of >0.25 mmol/L (compared with control).Nonresponders were those in whom the TG variation was within the±0.25 mmol/L margin. Generalized linear models were conducted todetermine the predictors of the TG response to DHA and EPA.

Results: There was a greater proportion of responders after DHAthan after EPA (45% vs. 32%), thus resulting in a greater overallreduction in TG with DHA than with EPA (–13.3% vs. –11.9%,P < 0.01). The variation in TG was within the usual variation range in47% of participants after DHA and in 57% of participants after EPA. Atotal of 8% and 11%of individuals had an increase inTG thatwas greaterthan usual variation after DHA and EPA, respectively. Responders toDHA and to EPA had higher body mass index, waist circumference,and TG and insulin concentrations after the control phase comparedwith nonresponders (all P < 0.05).

Conclusions: Although high-dose DHA supplementation lowersserum TG concentrations in a greater proportion of individuals thanthe corresponding dose of EPA,<50% of individuals can be consideredresponders to either supplement. Men and women with a high bodymass index, waist circumference, and TG and insulin concentrationsare more likely to be responders than nonresponders to either DHA orEPA.

Funding SourcesFinancial support for this randomized controlled trial was provided

by a grant from the Canadian Institutes for Health Research (CIHR,MOP-123,494). Douglas Laboratories provided the EPA, DHA andcontrol capsules used in this study. JA is a recipient of a PhD Scholarshipfrom the Canadian Institutes of Health Research (CIHR) and the Fonds


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de recherche du Québec—Santé (FRQ-S). CV is supported by theEuropean Marie Skłodowska-Curie Actions.

Urinary Medium- and Short-Chain Acylcarnitines Sign theImpact of High Caloric and High Protein Diet on Fatty AcidMetabolism (OR17-07)

Daniel Tomé,1 Annemarie Rietman,2 Nadezda Khodorova,1Douglas Rutledge,1 Jessica Schwarz,2 Julien Piedcoq,1 SergePilard,3 Els Siebelink,2 Frans J Kok,2 MarcoMensink,2 andDalilaAzzout-Marniche1

1AgroParisTech; 2WUR; and 3Université de Picardie

Objectives: Increasing the protein content of a high-fat hypercaloricdiet was shown to reducing intrahepatic lipids and body fat mass.This study aimed to identify, in urinary metabolic profiles, biomark-ers associated to the consumption of a high-protein, hypercaloricdiet.

Methods: In a crossover design, 17 healthy subjects were randomlyassigned to consume a high-protein diet (HP; 37.7% of total energyfrom fat, 25.7% from protein, and 36.6% from carbohydrates; totalenergy 2+ MJ/d) and a normo-protein diet (NP; 39.4% of total energyfrom fat, 15.4% from protein, and 45.2% from carbohydrates; totalenergy 2+ MJ/d) (Figure 1). A control group was included (CDgroup; n = 10; 27.8% of total energy from fat, 16.9% from protein,and 55.3% from carbohydrates). After 2 wk of adaptation, 24-h urinesamples were collected after each period and [U-13C]palmitate wasadded for extrinsic isotopic labeling of meal lipids the day of a mealchallenge. A nontargeted metabolomic profiling was done on the urinesamples using ultrahigh-performance liquid chromatography-QTOF-MS and 13C enrichment was determined using isotopic ratio massspectrometry. Independent components-discriminant analysis (IC-DA)using the JADE algorithm was applied to the data. Permutation testswere used to validate the observed models and the differences wereconfirmed by ANOVA.

Results: The IC-DA enabled us to distinguish the hypercaloricgroups from the CD group (Figure 2A) owing to the increased excretionof 2 medium-chain acylcarnitines (10:3 and 8:1-carnitines; P < 0.05),whatever the protein intake and the intervention period. The HPgroup was distinguished from the CD and NP groups (Figure 2B)owing to the increased urinary excretion of carnitine and 4 short-chain acylcarnitines (C2, C3, C4, and C5 acylcarnitines; P < 0.05),and decreased urinary excretion of heterocyclic nitrogen-containingmetabolites (oxoguanine and uric acid; P < 0.05). Moreover, the 13Cisotopic enrichment of urines from the HP group was higher than fromthe NP and CD groups (P < 0.0001), suggesting an increase in fatexcretion with HP feeding.

Conclusions: The increased urinary excretion of medium-chainacylcartines could be biomarkers of a hypercaloric diet and hepatic lipidaccumulation, whereas short-chain acylcarnitines could be biomarkersof an HP diet, together with elimination of excess fat and reducedhepatic fat accumulation.

Funding SourcesAgroParisTech, INRA, WUR, and the Dutch Dairy Association.

Essential Fatty Acids Linoleic Acid and α-Linolenic Acid Sex-Dependently Affect Glucose Homeostasis in Obese Humans andMice (OR17-08)

Pan Zhuang,1 Jingjing Jiao,2 and Yu Zhang1

1Zhejiang University, China; and 2Zhejiang University School ofMedicine, China

Objectives: The aims of this study were 1) to assess the long-termassociations between intake of linoleic acid (LA) and α-linolenic acid(ALA) and the risk of type 2 diabetes (T2D) in a population-basedcohort, and 2) to investigate the mechanism of action based on gutmicrobiota in a high-fat diet–induced obese (DIO) mouse model.

Methods: The occurrence of T2D in 15,100 apparently healthyChinese adults from the China Health and Nutrition Survey (CHNS,1997–2011) was followed up for a median of 14 y. Diet was assessedbased on 3-d 24-h recalls with a weighing technique in survey years1997, 2000, 2004, 2006, 2009, and 2011. Cumulative averages of dietaryintakes were calculated to represent long-term diet and minimizewithin-person variation. HRs and 95% CIs were estimated using theCox proportional hazards regression model. A high-fat diet was fed tomice for 10 wk to produce the obese mouse model. The DIOmice werethen fed with a high-fat diet (HFD) or HFD enriched with LA or ALA(0.2% wt:wt) for 15 wk to investigate the effect of the diet on glucosehomeostasis, gut microbiota composition, and adipose inflammation.

Results: The prospective cohort revealed that ALA intake wasassociated with lower incidence of T2D in obese and overweightmen (HR comparing extreme quartiles: 0.55; 95% CI: 0.32, 0.93; P-trend = 0.02) and women (HR comparing extreme quartiles: 0.53;95% CI: 0.34, 0.85; P-trend = 0.007), whereas LA intake was inverselyrelated to T2D risk only in obese and overweight women (HRcomparing extreme quartiles: 0.56; 95% CI: 0.36, 0.89; P-trend = 0.01).LA administration ameliorated insulin resistance in female DIOmice and ALA improved glucose homeostasis in male DIO mice.Meanwhile, significant sex-dependent changes of gut microbiota weredetected after LA or ALA intervention. Endotoxemia and systematicand adipose inflammation were relieved in ALA-fed male mice andLA-fed female mice. Although the expression of GLUT4 was notaffected, LA (for female) and ALA (for male) supplementation signif-icantly elevated phosphorylation of Akt, indicating enhanced GLUT4translocation.

Conclusions: Long-term intake of dietary LA (for females) andALAmay produce favorable effects on glucose homeostasis in already obeseindividuals. These effects may result from the sex-specific alterationsof intestinal microbiota that relieve adipose inflammation through thegut-adipose axis.

Funding SourcesThis work was supported by the National Natural Science Founda-

tion of China (31,401,659, 31,201,307, and 81,300,309).


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FIGURE 1 ALA and LA affect obesity in DIO mice.


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FIGURE 2 ALA and LA impact glucose homeostasis in DIO mice.


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FIGURE 3 ALA and LA sex-dependently alter gut microbiota composition.


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Associations of ALA and LA intakes with T2D risk in CHNS.


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Resistant Maltodextrin Produces a Greater Change in FecalBifidobacteria Counts Compared with Placebo: A Double-Blind,Randomized, Crossover Study (OR29-01)

Alyssa M Burns,1 Rebecca Solch,1 Jennifer Dennis-Wall,1 MariaUkhanova,1 Carmelo Nieves,1 Volker Mai,1 Mary Christman,2Dennis T Gordon,3 and Bobbi Langkamp-Henken1

1University of Florida; 2MCC Statistical Consulting LLC; and3ADM/Matsutani, LLC

Background: Dietary fiber stimulates the growth of potentiallybeneficial bacteria (e.g., bifidobacteria), yet daily fiber recommen-dations are not met by most Americans. The addition of resistantmaltodextrin (RMD) to the diet may help individuals meet their fiberrecommendations; however, the effect of RMD on potentially beneficialbacteria has not been well established.

Objective: The primary objective of this study was to determine theeffect of adding 0, 15, and 25 g RMD to the diets of healthy free-livingadults on fecal bifidobacteria counts.

Methods: In this double-blind, placebo-controlled, 3-interventioncrossover study, 51 participants (age: 26.3 ± 1.0 y) were randomized toconsume 0, 15, or 25 g RMDdaily for 3 wk, followed by a 2-wk washout.Participants collected total stools for 2 d at weeks 0 and 3 of each inter-vention for stool wet weight (WW) measurements and fecal bifidobac-teria counts, which were obtained by quantitative polymerase chainreaction.Weekly 24-h dietary recalls were completed to assess total fiberintake.

Results: Compared with 0 g, 25 g of RMD resulted in agreater change in fecal bifidobacteria (–0.17 ± 0.09 vs. 0.17 ± 0.09log10(counts), respectively; P = 0.0084). Stool WW increased with 25g of RMD (150 ± 111 g/d vs. baseline 121 ± 11 g/d; P = 0.0106). Totalaverage fiber intakewas significantly higher with 15 g (18.0± 0.6 g/1000kcal) and 25 g (25.3 ± 1.1 g/1000 kcal) compared with 0 g of RMD(8.4 ± 0.4 g/1000 kcal), and exceeded current recommendations of 14g · 1000 kcal–1 · d–1.

Conclusions: Average daily total fiber intakes exceeded recom-mendations with both 15 and 25 g of RMD. Additionally, 25 g ofRMD increased fecal bifidobacteria counts and stool WW, whichsuggests potential health benefits associated with increased total fiberintake.

Funding SourcesADM/Matsutani, LLC.

The Soluble Fiber α-Cyclodextrin Does Not Increase the FecalLosses of Dietary Fat: A Double-Blind, Randomized, Placebo-Controlled, Crossover Trial (OR29-02)

Kelli A Lytle, Deb Harteneck, and Michael D Jensen

Mayo Clinic, MN

Background: Supplemental α-CD, a soluble dietary fiber, may beeffective in mitigating metabolic syndrome by improving plasma lipidprofiles and promoting weight loss. Preliminary evidence suggeststhat it may exert these effects by binding dietary fatty acids andreducing their absorption; however, this has not been tested inhumans.

Objective: The primary objective of this study was to test whethersupplemental α-cyclodextrin (α-CD) increases fecal content of dietaryderived lipid in healthy humans.

Methods:This was a randomized, double-blind, placebo-controlled,2-way crossover study. A group of 8 volunteers underwent 2 separatestudy visits with or without α-CD supplementation, with a washoutperiod of ≥2 wk. On the first morning of each visit, the volunteersconsumed a standardized liquid breakfast containing [14C]tripalmitinand [3H]triolein with 2 g of α-CD or placebo, followed by the α-CD or placebo at each subsequent meal for 2 d. Plasma sampleswere drawn every 60 min for 360 min following ingestion of thelabeled meal to measure total triglyceride (TG) and glucose concen-trations, plus appearance of tracer in plasma TGs. The volunteersconsumed a diet containing 100 g/d of fat. All feces were collectedfor 72 h following administration of dietary radiotracers with andwithout α-CD and assayed for radiotracer content and total fecalfat.

Results: We found virtually no 3H radiotracer in fecal lipidswith either α-CD or placebo, but an average of ∼20% of the 14Ctracer was recovered in fecal lipids, with no difference between α-CDand placebo. There was also no difference in total fecal fat content,appearance of radiotracers in postprandial plasma TGs, or absolutepostprandial TG or glucose concentrations between α-CD and placebo.Of interest, the plasma [14C]triglyceride response was less than theplasma [3H]triglyceride response, and the losses of 14C in fecal lipidswere positively correlated with total fecal fat.

Conclusions:α-CD supplementation did not increase loss of dietarylipid in stool or total fecal fat. The greater loss of 14C in stoolfrom [14C]tripalmitin, combined with the lesser appearance in plasma[14C]triglyceride, implies significantly different metabolic handling ofthese 2 dietary fat tracers.

Funding SourcesFunding for this research was provided by SFI Research Party Ltd.

Preloading Different Kinds of Coffee before a High Glycemic–Index Breakfast Led to Differences in Postprandial Glycemic andInsulin Responses (OR29-03)

Hon Ting Tommy Wong, Jennifer Man Fan Wan, and JimmyChun Yu Louie

University of Hong Kong

Objective: The aim of this study was to assess the effect of drinkingdifferent types of coffee before a high glycemic index (GI) breakfast onpostprandial glucose and insulin concentrations.

Methods: In this randomized, crossover, acute feeding study, 21apparently healthy, normal-weight adults completed 12 study sessions.In each session, fasted subjects first drank one of the following testdrinks (with low-fat milk and sugar): espresso coffee, instant coffee,boiled coffee, decaffeinated coffee, and water; and control (plain water).After 60 min, they ate a high-GI standard breakfast made of rice cereal,rice milk, and glucose powder. Glucose and insulin concentrationswere measured from capillary blood samples before drinking the testdrinks (–60 min), before eating the breakfast (0 min), and at 15, 30,45, 60, 90, and 120 min thereafter. Each test drink was given twice,and the test sessions were carried out ≥3 d apart. The incremental areaunder the curve (iAUC) for glucose and insulin was calculated using


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the trapezoidal rule. Differences in postprandial glucose (ppBGL) andinsulin concentrations and their iAUCs between test drinks were testedfor statistical significance using paired-sample t tests.

Results: In all sessions, the peak glucose concentration appeared at30 min, and no significant difference was observed between groups.Between 30 and 90 min, the glucose concentration decreased in allsessions, with the drop in ppBGL being the slowest in the controlsessions—the mean difference in glucose concentration from thebaseline of the control was highest at 60 (17–50% higher than all othertest drinks, P < 0.05 except for boiled coffee) and 90 min (18–40%higher than all other test drinks, P < 0.05 except for espresso andinstant coffee). Insulin concentrations increased from 0 to 30 min in allsessions, with that of the control rising the slowest—themean differencein insulin concentration from the baseline of the control was the lowestat 15 (22–41% lower than all other drinks, all P< 0.05) and 30min (12–29% lower than all other drinks,P< 0.05 for espresso and boiled coffee).Glucose and insulin iAUCs were not significantly different between alltest drinks.

Conclusion: Drinking coffee with low-fat milk and sugar beforeeating a high-GI breakfast caused the ppBGL to drop faster whencompared with drinking water. This may be because of a more rapidinsulin response, rather than an overall increase in insulin secretion.

Funding SourcesFunded by an internal research grant from the University of Hong

Kong.

Addition of Orange Pomace Alters Acute Glycemic Response toOrange Juice in Healthy Adults (OR29-04)

Indika Edirisinghe,1 Yancui Huang,1 Eunyoung Park,1 RebeccaReplogle,2 Thomas W Boileau,2 Jin-E Shin,2 Britt Burton-Freeman1

1Center for Nutrition Research, Institute for Food Safety andHealth,Illinois Institute of Technology; and 2PepsiCo R&D, IL

Objectives: Orange pomace is a by-product of orange juiceproduction and is a rich source of fiber. The goal of the present studywas to determine the impact of the addition of orange pomace to orangejuice on postprandial glycemic response.

Methods: A group of 10 healthy subjects (age: 27.9 ± 7.7 y;body mass index: 22.1 ± 1.1 kg/m2, mean ± SD) participated in arandomized, 2-arm, crossover clinical trial to test the glycemic responseto 252 g of 100% orange juice (OJ) or 100% orange juice with 5 g oforange pomace fiber (OPF; 250 g total beverage weight). The fiber levelwas chosen because it is similar to the amount found in an equivalentweight of whole orange fruit. Treatments were matched for availablecarbohydrates (OPF and OJ, 24.2 g). Blood samples were collected andglucose and insulin concentrations were measured at fasting (0 min)and every 15 min for 2 h after consuming the study juice products.

Results: Analysis of the 2 h incremental area under the curve(iAUC0–2 h) indicated a significant reduction in blood glucose afteringesting theOPF juice (243.7± 229.5mg ·min–1 ·dL–1) comparedwiththe OJ juice (872.1 ± 275.5 mg · min–1 · dL–1), P = 0.02. Peak glucoseconcentrations were also lower after the OPF juice compared with theOJ juice (116.2 ± 14.7 vs. 129.5 ± 15.6 mg/dL, respectively; P< 0.001).No significant difference was observed in insulin responses betweenOPF and OJ treatments, respectively, as measured by iAUC (iAUC0–2 h:

988.26 ± 199. vs. 986.79 ± 175.9 μIU · min–1 · mL–1; P = 0.9892) orpeak insulin (Cmax: 26.9 ± 13.5 vs. 30.6 ± 16.5 μIU/mL; P = 0.202).

Conclusions: This study demonstrated that adding 5 g of fiber fromorange pomace to 250 g of orange juice attenuated the postprandialglucose response without impacting insulin.

Funding SourcesPepsiCo, Inc.

Postprandial Blood Glucose, Insulin and Gastrointestinal Hor-mone Response to Starchy Foods (OR29-05)

Coby Eelderink, Roel Vonk, and Marion Priebe

UMCG, Netherlands

Objective: Frequent high blood glucose concentrations after theconsumption of starchy foods are implicated in the development of type2 diabetes (T2D). Decreasing the glycemic and insulinemic responsesmight be beneficial for both the prevention and the management ofthe disease. The aim of the study was to elucidate the underlyingmechanisms by investigating detailed glucose kinetics and metabolicresponses of starchy foods with varying food structures.

Methods: A cohort of 10 healthy male volunteers consumeddifferent 13C-enriched wheat products, while receiving a primed-continuous D-[6,6-2H2]glucose infusion. The dual-isotope techniqueenabled the calculation of glucose kinetics: rate of appearance ofexogenous glucose (RaE), endogenous glucose production, and glucoseclearance rate (GCR). In addition, postprandial plasma concentrationsof glucose, insulin, glucose-dependent insulinotropic polypeptide(GIP), glucagon-like peptide-1 (GLP-1) and bile acids (BAs) wereanalyzed. Fiber-rich wheat products were prepared with differentstructural features (pasta, bread with 85% broken wheat kernels, flatbread, control bread).

Results: The digestibility of starch is not always reflected by theglucose response. Pasta, with a slow appearance of starch-derivedglucose, resulted in a similar high glycemic response as control bread,but with a lower insulinemic response. A similar response was observedafter the consumption of compact flat bread. Digestibility of starch, asreflected by the RaE, correlated very well with GIP (r = 0.71–0.84),in contrast to GLP-1 (r = 0.47–0.70); the GLP-1 response was alsoinfluenced by the structure of the bread (presence of broken wheatkernels). BAs may play a role in this regard. The influence of theincretins, mainly GIP, on insulin concentrations and thus also on GCRcould explain the total glucose responses.

Conclusions: Slower intestinal uptake of glucose froma starchy foodcan result in lower postprandial insulin andGIP concentrations, but notnecessarily in a lower glycemic response, because of a slower GCR. Evenwithout reducing postprandial glycemia, productswith slowly digestiblestarch can have beneficial long-term effects. Starchy food structure andthus starchy food processing can affect the metabolic outcome, and istherefore an intriguing target for the prevention of T2D.

Comparison of Diets Enriched in Simple, Refined, or UnrefinedCarbohydrate onSubcutaneousAdiposeTissue InflammatoryMark-ers and Serum Cardiometabolic Risk Indicators (OR29-06)

Huicui Meng,1 Nirupa Matthan,1 Susan K Fried,2 SilviaBerciano Benitez,1 Maura Walker,2 Jean Galluccio,2 and Alice HLichtenstein2


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1Jean Mayer USDA Human Nutrition Research Center on Aging atTufts University, MA; and 2Icahn School of Medicine at Mount Sinai,NY

Objective: Data for direct comparisons between different typesof carbohydrate (simple, refined, and unrefined) on cardiometabolicrisk indicators are limited. Our aim was to determine the relativecomparability for an isocaloric exchange of simple carbohydrate(SC; e.g., sucrose, high fructose corn syrup), refined carbohydrate(RC; e.g., white rice, bread or pasta made from white flour) andunrefined carbohydrate (UC; e.g., brown rice, bread or pastamade fromwhole wheat flour) on adipose tissue inflammatory markers, ex vivocholesterol efflux of peripheral blood mononuclear cells (PBMC), andserum cardiometabolic risk indicators.

Methods: Participants [postmenopausal women and men (n = 11),aged 65 ± 8 y, with a body mass index of 29.8 ± 3.2 kg/m2 andLDL cholesterol ≥2.6 mmol/L] were provided with each of 3 diets(60% of total energy from carbohydrate, 15% from protein, 25%from fat) for 4.5 wk using a randomized crossover trial with 2-wkwashout periods between phases. The variable component was anisocaloric exchange of foods containing SC, RC, or UC. Body weightwas maintained within ±2 kg. Abdominal subcutaneous adipose tissuewas aspirated to assess macrophage and inflammatory marker geneexpression, ex vivo cytokine secretion, and adipocyte area; PBMCswereisolated to assess ex vivo cholesterol efflux; and serum lipid, lipopro-tein, glucose, insulin, and inflammatory marker concentrations weredetermined.

Results: Adipose tissue gene expression and ex vivo cytokinesecretion were similar between diets. Mean adipocyte area was largerand percentage of large adipocytes (area >1800 μm2) was higher atthe end of the RC- compared with the SC- and UC-enriched dietphases (P< 0.01). Fasting and postprandial serum cardiometabolic riskindicators and ex vivo cholesterol effluxwere similar between diets, withthe exception of higher fasting serum LDL- and non-HDL cholesterolconcentrations after the RC- compared with the SC- and UC-enricheddiets (P < 0.01).

Conclusions: Consuming a diet enriched with RC compared withSC or UC resulted in a larger mean abdominal subcutaneous adipocytearea and higher fasting serum LDL- and non-HDL cholesterol concen-trations, but had little effect on other cardiometabolic risk indicators.This study raises the intriguing possibility that RC may have uniqueadverse effects on cardiometabolic risk indicators distinct from SC andUC.

Funding SourcesThis work was supported by the USDA, under agreement No. 58-

1950-4-401, NHLBI T32-HL069772 and pilot funds from the JeanMayer USDA Human Nutrition Research Center on Aging at TuftsUniversity and the Boston Obesity Research Center.

Daily Aspartame Consumption Has No Effect on Glycemia,Appetite or BodyWeight in Healthy Adults (OR29-07)

Richard D Mattes,1 Kelly Higgins,1 and Robert Considine2

1Purdue University, IN; and 2Indiana University School of Medicine

Objectives: One goal of low-calorie sweeteners use is to reducesugar intake and postprandial increases in glycemia. Acute feeding trials

have yielded inconsistent findings on these outcomes. The present trialevaluated the effects of daily aspartame ingestion for 12 wk on glycemiaas a primary outcome and appetite and body weight as secondaryoutcomes.

Methods: A sample of 100 healthy adults were randomly assignedto consume 0, 350, or 1050 mg of aspartame daily for 12 wk.Baseline measurements included body weight, body composition,heart rate, urinary creatinine and para-aminobenzoic acid (PABA),24-h appetite, and an oral glucose tolerance test with concurrentmeasurement of insulin, glucagon-like peptide-1 (GLP-1), glucose-dependent insulinotropic polypeptide (GIP), and leptin. The aspartamewas delivered via a fruit-flavored beverage and capsules including PABAto permit assessment of treatment compliance. All participants reportedto the laboratory weekly to obtain new supplies and for measurementof body weight, blood pressure, heart rate, and waist circumference.At weeks 4, 8, and 12, 24-h urine samples and 24-h appetite logs werecollected. At the end of 12 wk, baseline measurements were repeated.

Results: No significant differences were observed for blood glucoseor insulin, resting leptin, or postprandial GLP-1 or GIP concentrationsbetween treatment groups over the 12-wk intervention. There also wereno treatment effects of aspartame ingestion on appetite, body weight,or body composition over the trial. PABA analyses indicated that meancompliance with aspartame ingestion was ∼95%.

Conclusions: Daily ingestion of aspartame at doses approximatelyequal to the population mean and the 95th percentile for 12 wk had nosignificant effect on glycemia, appetite, or bodyweight in healthy adults.

Funding SourcesAjinomoto Co., Inc.

Consumption Effects of 5 Sweeteners Taken for 12 wk on BodyWeight, Energy Intake, and Energy Expenditure (OR29-08)

Kelly A Higgins and Richard Mattes


Objectives: The purpose of the research was to compare the effectsof consumption of 4 low-calorie sweeteners (LCSs) and sucrose on bodyweight, energy intake, and energy expenditure over a 12-wk beverageintervention in adults (age: 18–60 y) with overweight or obesity (bodymass index between 25 and 40 kg/m2).

Methods: In a parallel-arm design, 155 participants were randomlyassigned to consume 1.25–1.75 L of beverage sweetened with 1 of5 sweeteners [sucrose, aspartame, saccharin, sucralose, rebaudiosideA (reb A)] daily for 12 wk. The beverages contained either 400–560kcal/d (sucrose treatments) or <5 kcal/d (LCS treatments). Baselinemeasurements included body weight, energy intake with three 24-hdietary recalls, and energy expenditure with triaxial accelerometers.Body weight was measured every 2 wk; energy intake and energyexpenditure were assessed every 4 wk. Also every 4 wk, 24-h urinecollections were completed to determine study compliance via para-aminobenzoic acid excretion.

Results: The data from this trial suggest that there are significantlydifferent changes in bodyweight between the sweetener treatments overthe 12-wk trial. Sucrose and saccharin consumption led to increasedbody weight across the 12-wk intervention (� weight = +2.45 and+1.170 kg, respectively; P ≤ 0.008), but were not significantly different


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from each other. Although there was no significant change in bodyweight with consumption of the other LCS treatments comparedwith baseline, changes in weight (sucrose – LCS) were significantlydifferent between sucrose and aspartame, reb A, and sucralose after12 wk (weight difference = 1.12, 1.26, and 2.66 kg, respectively;P ≤ 0.028). In addition, change in body weight was significantly lowerbetween sucralose and all other LCSs (weight difference ≥1.39 kg;P = 0.007). Energy intake and energy expenditure analyses acrosstreatment groups throughout the trial are underway. Compliance forall beverage treatments was ∼72%, with no significant difference incompliance between groups.

Conclusions: Sucrose and saccharin consumption significantlyincrease body weight compared with aspartame, reb A, and sucralose.All LCSs contribute negligible energy, but differ in their effects on bodyweight.

Nanoliposome-Mediated Delivery of the Antidiabetic ProteinE4orf1 (OR42-01)

Zahra Feizy, Swetha Peddibhotla, Shu Wang, Vijay Hegde, andNikhil Dhurandhar

Texas Tech University

Objectives: E4orf1 is an adenoviral-derived protein that promotescellular glucose uptake by upregulating AKT phosphorylation. Proof-of-concept studies show that the transgenic or viral vector–mediatedexpression of E4orf1 remarkably enhances glycemic control in animalmodels. However, a system suitable for delivering the E4orf1 proteinin humans is needed to extend the findings to a clinical setting. As thefirst step, our goal here was to show the construction and testing ofnanoliposome, a well-known nanoparticle, as a carrier to deliver theE4orf1 protein to 3T3-L1 cells in vitro.

Methods: Glutathione S-transferase (GST)-tagged E4orf1 proteinwas encapsulated in nanoliposomes, which were prepared usingSoy phosphatidylcholine and labeled with Rhodamine-PE (phospho-ethanolamine). The size and polydispersity index (PI) of nanoliposomeswere measured using a Brookhaven BI-MAS particle size analyzer. Totest the delivery efficacy of the nanoliposomes, murine 3T3-L1 cellswere treated with E4orf1-containing nanoliposomes (E4 group) or voidnanoliposomes (Void group), and expression was determined over timeby immunofluorescence. Cell lysates from treated cells were used toexamine changes in molecular signaling.

Results: The diameter of nanoliposomes was 133.5 and 90.7 nmand the PI was 0.2 and 0.1 for the E4 and Void nanoliposomes,respectively. The average encapsulation efficiency was 99% as measuredby enzyme immunoassay using anti-E4orf1 antibodies. The E4 groupof cells showed maximum GST and Rhodamine expression at 24 hcompared with 2, 4, and 12 h. The Void group showed the expression ofRhodamine, but not GST. The expression of E4orf1 in the E4 group ofcells was confirmed at 24, 48, and 72 h. Western blot analysis in cellstreated for 24, 48, and 72 h showed a significant increase in p-AKTexpression at 72 h in E4 cells compared with the Void group.

Conclusions: We report the first successful delivery of E4orf1encapsulated in liposomes and the expected effect of E4orf1 on cellsignaling. It provides the proof of concept for extending the use of ananoparticle-mediated delivery system to investigate the antidiabeticpotential of E4orf1 in humans.

Funding SourcesTexas Tech University research start-up.

Very Low Density Lipoprotein Receptor Mediates Obesity-Induced Insulin Resistance and Oxidative Stress in Skeletal Muscle(OR42-02)

Tahar Hajri, Viviana Angamarca, and Luis Caceres

Hackensack University Medical Center, NJ

Background: Obesity is associated with excess lipid depositionin skeletal muscle, leading to a state of lipotoxicity characterizedby increased oxidative stress and insulin resistance. Very low den-sity lipoprotein receptor (VLDLR), a member of the LDL receptorfamily, binds apolipoprotein E-triglyceride (TG)-rich lipoproteins andincreases the catabolism of TG-rich lipoprotein. Although VLDLR ishighly expressed in skeletal muscle, its role in muscle lipotoxicity is stillunclear.

Objective: This study was conducted to determine the role ofVLDLR in obesity-induced lipotoxicity in skeletal muscles and culturemyocytes.

Methods: VLDLR expression, oxidative stress parameters, anduptake ofVLDL-TGuptakewas studied inC2C12myocytes and skeletalmuscles of lean control (ob+/–), obese leptin-deficient (ob/ob), andleptin-VLDLRdouble null (ob/ob-vldlr–/–)mice. In addition, regulationof insulin sensitivity, glucose uptake, and superoxide production wereinvestigated in C2C12 myotubes.

Results: Compared with lean control mice, VLDLR expressionwas about 2-fold higher in skeletal muscles, including soleus andgastrocnemius, than obese ob/ob mice. This was associated withincreased muscle lipid content and enhanced VLDL lipids uptakefollowing in vivo injection of [3H]TG-labeledVLDL.VLDLRdeficiencyin ob/ob mice reduced the lipid content in skeletal muscles anddecreased the VLDL-lipid uptake, while enhancing insulin sensitivityand glucose uptake. Lipid peroxides and hydrogen peroxides, both ofwhich are oxidative stress markers, and the expression of NADPHoxidase (Nox) were markedly higher in skeletal muscles of ob/ob mice.These parameters were normalized in ob/ob-vldlr–/– mice in associationwith a reduction of Nox-dependent superoxide production.

Conclusions: In C2C12 myotubes, small interfering RNA-mediatedknockdown of VLDLR expression reduced intracellular lipid depo-sition, and prevented VLDL-induced insulin resistance and excessproduction of Nox-dependent superoxides.

Funding SourcesHackensack Medical Center.

Muscle Inflammation Is Upregulated during Energy Deficitat High Altitude and Associated with Increased Myogenesis andDecreased Fat-Free Mass Loss (OR42-03)

Stefan M Pasiakos,1 Lee Margolis,1 Claire Berryman,1 Christo-pher Carrigan,1 Nancy Murphy,1 John Carbone,2 Marcas MBamman,3 Arny Ferrando,4 and Andrew J Young1

1US Army Research Institute of Environmental Medicine; 2EasternMichigan University; 3University of Alabama, Birmingham; and4University of Arkansas for Medical Sciences


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Background:Transient elevations inmuscle inflammatory signalingafter exercise, particularly via the tumor necrosis factor (TNF)-likeweak inducer of apoptosis (TWEAK) and its receptor fibroblast growthfactor-inducible protein 14 (Fn14), is considered promyogenic, whereaschronic elevations inhibitmyogenesis and promotemuscle atrophy. Thecombined effects of physiologic and environmental stress on muscleinflammatory signaling remain unclear.

Objective: The aim of this study was to explore muscle inflam-mation, myogenesis, and proteolysis in 16 men at sea level (SL) andfollowing 21 d of energy deficit (–1862 ± 525 kcal/d) at high altitude(HA, 4300 m).

Methods:Total bodymass (TBM), fat-freemass (FFM), and fatmass(FM) were assessed using dual-energy X-ray absorptiometry. Geneexpression and proteolytic enzymatic activities were assessed in vastuslateralis samples collected in the basal state at SL and HA.

Results: Participants lost 7.2 ± 1.8 kg of TBM (P < 0.05); 43 ± 30%and 57 ± 30% of the TBM lost was FFM and FM, respectively. Fn14,TWEAK, TNF-α-receptor (TNFα-R), TNFα, MYOGENIN, and pairedbox protein–7 (PAX7) were upregulated, and proteolytic activity wassimilar at HA and SL (P < 0.05). Stepwise linear regression identifiedthat Fn14 explained the highest percentage of variance in FFM loss(r2 = 0.51; P < 0.05). Dichotomization of volunteers into HIGHand LOW Fn14 responders based on median Fn14 mRNA expressionindicated HIGH lost less FFM and more FM (28 ± 28% and 72 ± 28%,respectively) as a proportion of TBM loss than LOW (58 ± 26% and42 ± 26%; P < 0.05) at HA. MYOGENIN mRNA was also greater forHIGH versus LOW (P < 0.05).

Conclusion:These data suggest that heightened Fn14may attenuatemuscle loss associated with pronounced energy deficit at HA.

Funding SourcesSupported by US Army Medical Research and Materiel Command.

Roux-en-Y Gastric Bypass Cannot Change Methylation Levels ofSpecific Obesity-Related Genes (OR42-04)

Carolina Nicoletti Ferreira,1 Ana Crujeiras,2 Angel Diaz-Lagares,3 Felipe Casanueva,4 Marcela Pinhel,1 Bruno deOliveira,1 Vitor Pinhanelli,1 Julio Marchini,1 Wilson SalgadoJunior,1 and Carla B Nonino1

1Ribeirão Preto Medical School—University of São Paulo, Brazil;2Instituto de Investigaciones Sanitarias-Laboratorio 2 Complejo Hos-pitalario Universitario de Santiago (CHUS), Spain; 3Health ResearchInstitute of Santiago (IDIS) University Clinical Hospital (CHUS),Spain; and 4Department of Medicine, Endocrinology Section Santiago

de Compostela University Complejo Hospitalario Universitario deSantiago, Spain

Objectives: The aim of this study was to investigate how, and withwhat magnitude, Roux-en-Y gastric bypass (RYGB) modifies the DNAmethylation profile of obese women by performing a genome-wideepigenetic analysis in peripheral blood.

Methods: We undertook a prospective study with women from amixed ethnicity population. Using the Infinium Human Methylation450 BeadChip array, global methylation analysis was performed in24 severe obese patients [body mass index (BMI): >35 kg/m²; age:36.9 ± 10.2 y) before and 6 mo after RYGB, and 24 normal weightindividuals (BMI: 18.5–24.9 kg/m2; age: 36.9± 11.8 y). Anthropometryand body composition analysis were also evaluated. DNA qualitychecks, bisulfite modification, hybridization, data normalization, andstatistical filtering were performed. To identify consistent patterns ofdifferentially methylated CpG, a parametric t test was performed. Weapplied a threshold for the significant CpG sites based on �β , with aminimum value of 5% and a P-value of <0.01. All statistical analysiswas performed using R software (version 3.2.0).

Results: The RYGB procedure was able to promote a significantreduction in weight, waist circumference, and fat mass. In addition, 666CpG sites were changed by the surgical procedure. The average DNAmethylation levels of these sites increased after surgery (0.44 ± 0.13to 0.49 ± 0.12; P = 0.000). However, we found 544 CpG sitesthat were always differently methylated between obese and normalweight patients and were not influenced by RYGB (Figure 1). Thesedifferently methylated CpGs were associated with the Wnt and p53signaling pathways, and we highlighted the nicotinamide adeninedinucleotide (reduced form):ubiquinone oxidoreductase subunit S6(NDUFS6) and mitochondrial ribosomal protein L36 (MRPL36) genes.Also, we found that TMEM48 gene was positive correlated (baselinelevel) with the percentage of weight loss and the percentage ofBMI change. This CpG site showed low methylation levels in obesepatients, and those who had greater methylation levels lost moreweight.

Conclusion: RYGB promotes epigenetic changes of specific path-ways; however, a cluster of obesity-related genes remains unchangedeven after bariatric surgery. Thus, genome-wide DNA methylationanalysis suggests that 6 postoperative months is not sufficient time tomodify the epigenetic signature of obesity.

Funding SourcesSão Paulo Research Foundation (FAPESP) (grants #2017/0,7220-7,

#2016/0,5638-1 and #2015/18,669-0).


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FIGURE 1 Supervised clustering of the 544 CpGs that were foundto be differentially methylated between the presurgery obesepatients and normal weight women.

Weight Loss Downregulates CD36 Expression and ReducesInflammation Markers in Monocytes of Obese Subjects UndergoingBariatric Surgery (OR42-05)

Tahar Hajri, Toghrul Talishinski, Luis Caceres, Vivian Anga-marca, Ewing Douglas, Eid Sebastian, Novac Richard, andSchmidt Hans

Hackensack University Medical Center, NJ

Background:Cell differentiating-36 (CD36) is amultifunctional cellmembrane protein expressed on various cells, includingmonocytes andmacrophages. CD36 binds several ligands, including fatty acids (FAs)and oxidized lipids, thereby mediating their proinflammatory effects.

Objectives: In order to gain further insight into the role of CD36 inobesity-associated inflammation, we examined the expression of CD36in monocytes and adipose tissue macrophages monocytes of lean andobese subjects undergoing bariatric surgery.

Methods: Blood samples were collected from lean subjects [bodymass index (BMI): <25 kg/m2] and from obese subjects (BMI: >30kg/m2) before and after bariatric surgery. The concentrations of bloodlipids, inflammation markers, glucose, and insulin were assayed inplasma, and the homeostatic model assessment insulin resistance index(HOMA-IR) was determined. Blood monocytes were isolated by gra-dient centrifugation and CD14-positive magnetic activated cell sorting,and the expressions of CD36, interleukin-6 (IL-6), and tumor necrosisfactor α (TNF-α) were assayed byWestern blot and quantitative reversetranscriptase-polymerase chain reaction. In addition, we examined theuptake and effects of fatty acids on monocytes differentiation andphenotype expression.

Results: Bariatric surgery resulted in marked reduction of BMI,and this was associated with significant improvement of HOMA-IRand reduction of plasma free fatty acids (FAs), C-reactive protein,IL-6, and TNF-α. Compared with lean subjects, the expressions ofCD36, IL-6, and TNF-α were strikingly higher in monocytes ofobese subjects presurgery, but were significantly (P < 0.01) reduced12 mo postsurgery. Exposure of monocytes to palmitate enhanced theexpressions of IL-6, TNF-α, and CD11c, while reducing the expressionsof anti-inflammatory markers IL-4 and IL-10, and CD206.

Conclusions: The results indicate that the expression of CD36is upregulated in monocytes of obese subjects concomitantly withincreased inflammation markers. Weight loss downregulated CD36expression and promoted a shift of monocytes toward a lesserinflammatory phenotype. This effect is possibly related to the reductionof circulating fatty acids.

Funding SourcesHackensack University Medical Center.

Rats Resistant to High-Fat and High-Sucrose Diet-InducedObesity andMetabolic SyndromeHave aLowerDeNovoLipogenesisin Adipose Tissues and Liver (OR42-06)

Jean-Francois Huneau,1 Dominique Hermier,2 Olivier LMantha,2 Philippe Schmidely,3 Veronique Mathé,2 FrancoisMariotti,2 and Hélène Fouillet2

1AgroParisTech; 2UMR PNCA; and 3UMRMOSAR

Objectives: There is a high variability of metabolic response to ahigh-fat and high-sucrose diet, with individuals being either resistantor prone to develop obesity (O) alone or associated with a metabolicsyndrome (MS). This study aimed to compare de novo lipogenesis(DNL) between rats prone or resistant to diet-induced O and MS.

Methods:A total of 42maleWistar rats were fed a high-fat and high-sucrose diet for 4mo. Body composition andmetabolic parameters weremeasured at the end of the study and used to discriminate rats accordingto their sensitivity to O andMS, using unsupervised classification basedon 3 markers of O (weight, total and visceral adiposities) and 3 markersof MS (homeostatic model assessment insulin resistance index, plasmaand liver triglycerides). DNL was measured in the liver, epididymal(EAT) and subcutaneous (SAT) adipose tissues by deuterium tracing:after an initial intraperitoneal load (20 g/kg, 99.9 atom%), rats wereexposed to D2O (5 atom%) in drinking water for 2 d prior to killing,and D-enrichment was measured in tissue neutral lipids by elementalanalysis-isotopic ratio mass spectrometry and in plasma by GC-MS.In each tissue, the net flux of lipids synthesized by DNL and stored in


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situ (net DNL, mg/d) was estimated using an equation derived from aprecursor-product model.

Results: The classification distinguished 3 groups of rats: thoseresistant to O and MS (R, n = 15), those prone to O but resistant toMS (O, n = 13), and those prone to both O and MS (OMS, n = 14).The relative dietary intake per gram of bodyweight was similar betweenthe groups. Only OMS rats had steatosis (hepatic lipid content of 13%vs. 8% for R and O rats), and hepatic DNL was 40% lower in R and Orats compared with OMS. In adipose tissues, DNL was 35% (SAT) and45% (EAT) lower in R rats compared with O, and 35% lower in O ratscompared with OMS (table).

Conclusion: In this model, about one-third of the rats develop ahealthy O and one-third an O with MS. Rats resistant to MS (R and Orats) are those able to limitDNL in the liver, and thismetabolic flexibilitymay account in part for their limited hepatic steatosis. Rats that arealso resistant to O (R rats) are those that can also limit DNL in adiposetissues.

Funding SourcesFunded by INRA Department AlimH.

The Metabolic Effects of Sucrose Overfeeding Differs Accordingto Dietary Protein and Fat Composition in Healthy Humans(OR42-07)

Anna Surowska,1 Prasanthi Jegatheesan,1 Vanessa Campos,1Jérémy Cros,1 Léonie Egli,1 Anne-Sophie Marques,1 PhilippeSchneiter,1 Karin Zwygart,2 Roland Kreis,2 Chris Boesch,2Nathalie Stefanoni,1 Valentine Rey-Roethlisberger,1 and LucTappy1

1University of Lausanne, Switzerland; and 2University of Bernand Institute of Diagnostic Interventional and Pediatric Radiology,Switzerland

Objective: Many animal and human studies have documentedthat fructose or sucrose overfeeding increases body weight, plasmatriglyceride, and ectopic lipids, and impairs glucose tolerance. Whetherthe metabolic effects of sucrose are modulated by the dietary intakeof other macronutrients remains unexplored. We hypothesized thatpostprandial whole-body protein synthesis would be higher and ectopiclipid deposition would be lower when sucrose overfeeding is associatedwith a high vs. low dietary protein intake.

Methods: A group of 12 healthy young males and females wereincluded in a randomized, controlled crossover trial. Participants werestudied after a 3-d control (C) diet providing 100% of daily energyneeds (starch 45%, sucrose 10%, lipid 33%, protein 12%), and after6-d high-sucrose hypercaloric diets containing 150% of total requireddaily energy [29% starch, 34% sucrose, 7% lactose, with either 25% lipidand 5% protein (LP) or 10% lipid and 20% protein (HP)]. Whole-bodyprotein synthesis ([13C]leucine), plasma hormones, and metaboliteswere monitored over 6 h after ingestion of C, LP, or HP test mealsproviding 40% of daily energy needs. Intrahepatic lipid deposition(IHCL) and intramuscular lipid deposition (IMCL) were measured (bymagnetic resonance spectroscopy) at 1600 h during each condition.

Results: Postprandial protein synthesis decreased from 12.1 ± 1.4to 8.1 ± 0.5 g/5 h in the LP condition, but increased from 9.5 ± 0.7to 23.9 ± 4.8 g/5 h in the HP (P = 0.059 time, P = 0.025condition, P = 0.013 interaction). Postprandial plasma glucose and

triglyceride concentrations were not significantly different with HP andLP, but plasma insulin, glucagon, and insulin-like growth factor-I weresignificantly higher with HP. Postprandial plasma fructose and lactatewere significantly lower with HP than with LP. LP and HP significantlyraise IHCL and IMCL vs. C, but the percentage increases were largerwith LP thanwithHP (IHCL: 642%± 105% vs. 224%± 39%, P< 0.001;IMCL: 124.0% ± 3.0% vs. 111.4% ± 2.6%, P = 0.004).

Conclusion: Sucrose overfeeding associated with a high proteinintake results in a higher postprandial protein synthesis and a higherhepatic fructose uptake, but lower hepatic and muscle lipid deposition,than when combined with a low protein intake.

Funding SourcesSwiss National Science Foundation (SNSF).

A Healthy Diet Low in Simple Sugars and Low in Fat DoesNot Prevent Severe Hypertriglyceridemia in Obesity and/or Type 2Diabetes (OR42-08)

Barbara C Hansen,1 Kai-Lin Catherine Jen,2 Samuel CGidanian,1 Jennifer D Newcomb,1 and Uddhav K Chaudhari3

1Univesity of South Florida; 2Wayne State University, MI; and3National Institute for Research in Reproductive Health, India

Objective: The Endocrine Society Guidelines (2012) have identifiedserum triglyceride concentrations of 150–199 mg/dL as mild orborderline high, 200–999 as high and >1000 mg/dL (11.2 mmol/L) assevere hypertriglyceridemia (HTG). Primary HTG is generally believedto be the result of genetic disorders in triglyceride metabolism, with“secondary hypertriglyceridemia” thought to be caused by a high-fat orhigh-sugar diet, obesity, and/or diabetes. The optimal animal model forexamining themechanisms underlyingHTG and potential therapeuticsis one that develops naturally and spontaneously, includes a lipid profilevery similar to humans, and can be studied longitudinally in order toidentify causal or risk factors for the development of HTG. AlthoughHTG in rodents has been induced experimentally with a high-fat orhigh-fructose diet, or with genetic modifications, these models do notreflect the human condition. The aim of this study was therefore to de-termine possible causal factors for HTG in a nonhuman primatemodel.

Methods: We maintained a colony of 263 nonhuman primates(rhesus monkeys) on the same low-fat diet for life (Primate Lab Diet5038; containing glucose + fructose + sucrose + lactose, togethertotaling <5% of the nutrients). We evaluated their TG concentrationslongitudinally.

Results:We found that HTG (TG>150mg/dL) developed naturallyand spontaneously. Despite the life-long constant “healthy” diet compo-sition, serum TG concentrations varied widely and 63.8% of this colonydeveloped HTG. Among the 86 monkeys with overt diabetes, 89.5%were significantly hypertriglyceridemic (TG >150 mg/dL), and amongthe 12with severeHTG (TG>1000mg/dL), all had overt type 2 diabetes(T2D). The following risk factors were selected to evaluate their rolesin initiating and exacerbating the significant rises in TG under thesesteady conditions: development of obesity (not involved), progressionof insulin resistance and significant hyperinsulinemia (involved only atthe earliest TG rise), and onset of T2D (HTG preceding the expressionof diabetes).

Conclusions: Because of the consistent environment and constantdiet composition, we can conclude that the naturally occurring HTG


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reported here is genetic, and that the HTG was highly associated withthe genetics of early β cell dysfunction and eventually overt T2D.

Funding SourcesNIH NIA HHSN263200800022C, the University of South Florida

Office of Research and Innovation, and the National Institute forResearch in Reproductive Health, Mumbai, India.

Evaluation of Polylactose as a Prebiotic Dietary Fiber (P10-001)

Breann E Abernathy and Daniel Gallaher

University of Minnesota—Twin Cities

Objectives: Polylactose is a novel dietary fiber, synthesized byextrusion of lactose. To evaluate its potential as a prebiotic, wedetermined its fermentability and its effects on adiposity and glycemiccontrol in a diet-induced obesity animal model.

Methods: A total of 72 male Wistar rats were fed normal fat (NF)or high fat (HF, 50% fat by kcal) diets containing various fibers (6%fiber of interest and 3% cellulose, by weight), including cellulose (NFCand HFC), polylactose (HFPL), matched lactose (HFML), matchedto the residual lactose in the HFPL diet, and 2 established prebioticfibers, polydextrose (HFPD) and fructo-oligosaccharides (HFFOS).During weeks 8 and 9 of feeding the experimental diets, rats underwentoral glucose, insulin (by intraperitoneal injections), and pyruvate (byintraperitoneal injections) tolerance tests. At the end of week 10, bloodand organs were harvested and cecal contents collected.

Results: There were no significant differences in final body weightsbetween the groups, nor did the average daily food intake differsignificantly between the HF-fed groups. Rats fed the HFPL diet hadimproved glycemic control when compared with the positive control(HFC) rats, as indicated by a strong trend for a lower area underthe curve for the oral glucose tolerance test (P = 0.051), and lowerblood glucose at 60, 90, and 120 min. There were no significantdifferences between the groups in the insulin and pyruvate tolerancetests. Epididymal fat padweight was significantly decreased in theHFPLrats comparedwith all the otherHF groups (P< 0.05), and did not differfrom the normal fat control (NFC). HFPL rats had greater cecal weight(empty) and their cecal contents had a lower pH when compared withall other groups, suggesting that polylactose is much more vigorouslyfermented than the other prebiotic fibers.

Conclusions: Polylactose is a vigorously fermentable fiber, sug-gesting that it will elicit a change in the gut microbiome. We havealso demonstrated that consuming polylactose, in the context of ahigh-fat diet, prevents the accumulation of body fat normally seenwith this diet, and also improves glycemic control. As these effects ofpolylactose were greater than those of 2 established prebiotics, fructo-oligosaccharides and polydextrose, this suggests that polylactose maybe a potent prebiotic.

Funding SourcesMidwest Dairy Association.

Co-occurrence of Cardiometabolic Disease Risk Factors amongAging Filipinas in the Context of the Nutrition Transition (P10-002)

Linda S Adair,1 Delia Carba,2 and Paulita Duazo2

1University of North Carolina at Chapel Hill; and 2University of SanCarlos, Philippines

Objective: The nutrition transition, marked by changes in dietarypatterns and physical activity, is associated with increases in obesity,hypertension, and diabetes inmany low- andmiddle-income countries,but recent secular and age trends in co-occurrence of these conditionshave not been well studied in these settings. The aim of this study wasto examine predictors of prevalence and patterns of co-occurrence ofoverweight and obesity (OW; body mass index: >25 kg/m2), a highwaist circumference (WC;>80 cm), diabetes, and hypertension amongFilipino women participating in the Cebu Longitudinal Health andNutrition Survey (CLHNS) from 1998 to 2016.

Methods: The CLHNS is a community-based study that recruited>3000 pregnant women in 1983–84 and has followed them in multiplesurvey rounds for >30 y. Data from the 5 most recent surveyrounds (1998–2016) included blood pressure (measured by trainedinterviewers) and diabetes status (self-reported in 1998 and 2002,fasting glucose in 2005, and glycated hemoglobin in 2012 and 2016).Hypertension and diabetes were defined according to InternationalDiabetes Federation guidelines. The women, who ranged in age from29 to 62 y in 1998, were grouped in each year according to the presenceor absence of OW, high WC, hypertension, and diabetes, and weused multinomial logistic regression to identify factors associated withmembership in each group.

Results: Comparing 1998 to 2016, we found that the occurrenceof having none of these conditions declined from 50% to 20%; being“metabolically healthy” (OW or high WC but no hypertension ordiabetes) declined from 26% to 17%, hypertension increased from 21%to 59%, and diabetes increased from 2% to 14%. On average across allyears, only about half of the women with hypertension were OW or hada highWC. In general, women in groups that included OWor highWCwere older, taller, frommore urbanized communities, and in householdswith greater assets. The lowest prevalence of these adverse conditionswas in rural, more disadvantaged women. Further analyses will explorethe role of diet and other lifestyle factors.

Conclusions: Urbanization and increasing socioeconomic statusare strongly related to rising prevalence and co-occurrence of car-diometabolic diseases as womenmove frommiddle to older adulthood.

Funding SourcesNIA R01 AG039443.

Low PlasmaHDLCholesterol Is Associated with Greater Risk forHeart Disease in Patients with Metabolic Syndrome (P10-003)

Hana M Alyousef

University of Connecticut

Background: Low plasma HDL cholesterol has been recognized asa biomarker for heart disease and diabetes. Low HDL cholesterol is alsoone of the characteristics of metabolic syndrome (MetS).

Objective: The aim of this study was to analyze whether low HDLcholesterol is associated with the biomarkers of inflammation andoxidative stress in individuals classified with MetS.

Methods: A total of 40 subjects with MetS [11 men, 29 women; age:52.4± 9.5 y; bodymass index (BMI): 32.3± 3.7 kg/m2] were evaluated.Anthropometric data [weight, height, BMI, waist circumference (WC),blood pressure (BP)], fasting plasma biomarkers [lipids, glucose,liver enzymes such as alanine aminotransferase (ALT) and aspartate


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aminotransferase, plasma insulin, and glycated hemoglobin], biomark-ers of oxidative stress and inflammation [C-reactive protein, tumornecrosis factor α, monocyte chemoattractant protein-1, interleukin(IL)-6, and IL-8], antioxidants [glutathione peroxidase, superoxidedismutase, total antioxidant capacity (TAC), and catalase], and thio-barbituric acid reactive substances (TBARS) as a marker of lipidperoxidation were measured.

Results: Participants had a mean BMI of 32.3 ± 2.7 kg/m2, placingthem in the obesity category. In terms of MetS characteristics, allsubjects (100%) fit the criteria for WC; 63% either had high systolicBP or high diastolic BP, or both; 70% were hyperglycemic; 48% hadelevated plasma triglycerides (TG), and 43% had low HDL cholesterol.We divided the subjects into 2 categories: Low HDL cholesterol (men<40 mg/dL and women <50 mg/dL) (n = 17) and Normal HDLcholesterol (men ≥40 mg/dL and women ≥50 mg/dL) (n = 23). Thosewith Normal HDL cholesterol had lower systolic BP (129.9 ± 9.7vs. 121.2 ± 14.2 mm Hg, P < 0.05), lower TG (115.5 ± 56.8 vs.166.6 ± 8.0 mg/dL, P < 0.05), lower TBARS (0.143 ± 0.027 vs.0.159± 0.025μM, P< 0.05) and higher TAC (2.09± 1.5 vs. 1.23± 0.9mM Trolox equivalents, P < 0.05). In addition, strong negativecorrelations were found between HDL cholesterol and each of theparameters WC (r = –0.418; P < 0.01), insulin (r = –0.413; P < 0.025),and ALT (r = –0.324; P < 0.05).

Conclusion:These results confirm that lowHDL cholesterol is asso-ciatedwith lower antioxidant capacity, dyslipidemias, and hypertension.

Funding SourcesTA Sciences.

Short- and Long-Term Effects of High Monounsaturated andHigh n–6 Polyunsaturated Fat Diets onMetabolic Syndrome in Rats(P10-004)

Kathleen V Axen,1 Jo Ann Brown,2 Kate Russell,1 KeerteshwryaMishra,3 Kadeem Thomas,1 and Kenneth Axen1

1Brooklyn College of the City University of New York; 2KingsbrookJewish Medical Center, Brooklyn NY; and 3Wayne State University, MI

Objective: Although monounsaturated fatty acids (MUFAs) andn–6 (ω-6) polyunsaturated fatty acids (PUFAs) can improve aspectsof metabolic syndrome, the effects of high intakes of these fatsremain unclear. Since individuals following low-carbohydrate dietsmayconsume large quantities of fat (>50%of total energy), we compared theshort- and long-term effects of high MUFA and PUFA diets on featuresassociated with metabolic syndrome in male Sprague-Dawley rats.

Methods: Equal groups of weight-matched rats consumed a 55% fat,15% starch diet comprised of olive (MUFA) or safflower oil (PUFA),whereas a control group consumed a low-fat diet (LF, 15% fat, 59%starch) ad libitum for 1 (n= 30) or 8 (n= 30 )wk. Body composition andplasma concentrations of glucose, insulin, and triglyceride were mea-sured after a 24-h fast or after 16 h of refeeding with 85% carbohydrate.

Results: After 1 wk, body weights and energy intakes of MUFAand PUFA rats were greater than those of LF rats (P < 0.05), but thepercentage body fat did not differ between the 3 groups. Liver lipidconcentration was higher in PUFA vs. LF rats (P < 0.005), whereasplasma concentrations of triglyceride were lower in PUFA vs. MUFA orLF rats in the fasted or refed states (P < 0.05). Groups did not differ in

fasted concentrations of glucose or insulin, and all had similar increasesin insulin in response to refeeding.

After 8 wk, total and percentage body fat were greater in MUFA andPUFA vs. LF rats (P< 0.01), and liver lipid concentration was higher inMUFA vs. PUFA or LF rats (P < 0.05). Plasma glucose concentrationsdid not differ between groups in the fasted or refed states; insulinconcentrations were higher in MUFA than PUFA or LF rats in thefasted state (P < 0.05), but were higher in PUFA than MUFA rats aftercarbohydrate refeeding (P < 0.05). Plasma triglyceride concentrationsdid not differ between groups in the fasted or refed states. Glucose andinsulin responses to an intraperitoneal glucose tolerance test at week 7did not differ between groups.

Conclusions: 1) The initial effects of the PUFA diet on liver orplasma lipid concentrations were transient, changing by week 8; 2) dietdid not affect plasma glucose concentrations in the fasted state, or inresponse to a glucose load or high carbohydrate refeeding; and 3)despiteequal gains in body fat by week 8, rats on the MUFA diet showedhigher liver lipid and fasted plasma insulin concentrations than PUFAor LF rats, indicating that the high MUFA, but not the high PUFA, dietpromoted the development of metabolic syndrome.

Funding SourcesProfessional Staff Congress Research Award 67,833-0045.

Obesity-Prone Carbohydrate-Sensitive Rats Have Increased Pan-creatic mRNA Amylase and Accelerated Oxidation of DietaryGlucose (P10-005)

Dalila Azzout-Marniche,1,2,3,4 Catherine Chaumontet,3 NadezdaKhodorova,1 Julien Piedcoq,1 Gilles Fromentin,3 Daniel Tome,1Claire Gaudichon,1 and Patrick Even5

1AgroParisTech; 2UMR PNCA; 3INRA; 4AgroParisTech, UniversitéParis-Saclay; and 5CNRS

Objective: It was previously shown that rats can be sensitive orresistant to obesity under a diet rich in carbohydrates and low in fat(“STARCH” diet). These rats were classified as “carbohydrate sensitive”(CS) or “carbohydrate resistant” (CR). After ingestion of a meal, CS ratshave a larger rate of carbohydrate oxidation than CR rats. The objectiveof this study was to understand the mechanisms responsible for thiseffect.

Methods: A total of 32 male Wistar rats were fed a “STARCH” dietfor 7 wk (Figure 1). Body weight and food intake weremeasured 3 timesa week. Body composition was measured by MRI at the beginning ofthe experiment (T0), and after 3 (T3) and 6 (T6) wk. According to theiradiposity gain during the first 3 wk, rats were classified as CR (n = 9)or CS (n = 7). At 3 wk, the rats were fed a meal (56 kJ) with eitherSTARCH or maltodextrine (MALTO) labeled with 13C. Carbohydrateoxidation was measured from O2 and CO2 in the expired gases; 13CO2

was measured by multiflow isotopic ratio mass spectrometry. At 6 wk,the rats were killed, portal and venous blood was collected, and bodycomposition was measured by dissection.

Results: Fat mass (and adiposity) at T3 of the STARCH period wassignificantly lower in CS than in CR rats (21.9 ± 1.09 vs. 37.8 ± 1.37)with no differences on body weight (261.4 ± 4.44 vs. 256.1 ± 6.39) andlean mass (247.6 ± 4.13 vs. 238.8 ± 5.93). Recovery of 13CO2 in theexpired gases (Figure 2) showed no differences between CR and CS ratsafter ingestion of the MALTO meals, whereas after the STARCHmeals


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the recovery of 13CO2 was significantly higher in CS than in CR rats,showing that the catabolism of dietary starch occurred more rapidly inCS rats. This phenomenon is probably because of greater productionof amylase by the pancreas, as confirmed by a higher level of mRNAencoding for amylase in the pancreas and significantly higher amylaseactivity in the jejunum (and to a lesser extent in the portal blood) of CSrats (Figure 3).

FIGURE 1 Study design. MTT, meal tolerance test.

FIGURE 2 Evolution of 13CO2 in expired gases in CR and CS rats after ingestion of a STARCH or MALTO test meal (*** CR STARCH < CSSTARCH, P < 0.001) (A). (B) Total 13CO2 recovery during 6 h after meal ingestion. (C) Same data as in (B) expressed as a percentage ofingested carbohydrate oxidized during 6 h after meal ingestion.

Conclusion: InCS rats fed a standard STARCHdiet, a larger amylasesecretion by the pancreas is probably responsible for the increased rateof glucose absorption and oxidation, leading in the long-term to a gainin fat mass, as observed in rats fed a high glycemic–index diet.

Funding SourcesAgroParisTech, INRA


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FIGURE 3 Expression of pancreatic amylase (A), immunohistochemistry of pancreatic amylase (B), and amylase activity (C) in the intestinaltract of CR and CS rats 2 h after ingestion of a calibrated meal.

Effect of Aryl Hydrocarbon Receptor Activity on Lipid Accumu-lation, Insulin Content, and Secretion of Clonal Pancreatic β Cells(INS-1) of Excess Nutrient? (P10-006)

Siyouneh Baghdasarian,1 Karel Erion,2 Barbara Corkey,3 JudeDeeney3

1Boston University; 2University of California Los Angeles; and3Boston University School of Medicine

Objective: The aryl hydrocarbon receptor (AHR) translocates tothe nucleus and binds to the AHR nuclear translocator to regulatebiological responses upon ligand activation. The aim of this studywas to measure the effects of activation or inhibition of AHR activityon basal and glucose-stimulated insulin secretion (GSIS) from clonalpancreatic β cells (INS-1) cultured under normal and glucolipotoxic(GLT) conditions (high glucose and fatty acid).

Methods: Insulin content and secretion were measured with aHomogeneous Time Resolved Fluorescence (HTRF) assay kit fromcisbio. INS-1 cells were cultured in RPMI media containing 5 and 11mM glucose. Cells were preincubated with a receptor agonist (FICZ)and antagonist (CH223191) for 96 h. Insulin secretion was measuredover 2 h and reported as nanograms per million cells. Intracellular lipidwas measured by fluorescence after Nile red staining.

Results: Incubation of INS-1 cells with 11mMglucose and fatty acidincreased lipid droplets and basal insulin secretion and inhibited GSIScompared with cells cultured in 4 mM glucose, characteristic of GLT.INS-1 cells cultured in 5 or 7 mM glucose and treated with the AHRagonist (1.25–10 nM) exhibited increased lipid without a significanteffect on insulin secretion. INS-1 cells cultured at 11 mM glucoseand treated with antagonist had decreased lipid content and improvedinsulin secretion compared with cells cultured in 11 mM glucose alone.

Conclusions: The AHR may play a role in the development of GLTin pancreatic β cells cultured in excess nutrients.

Funding SourcesFunded by the NIH.

Omega-3 Fatty Acid Intake at Breakfast Decreases Hunger andIncreases Postprandial Energy Expenditure in Normal Weight andOverweight or Obese YoungWomen (P10-007)

Jamie I Baum,1 Eva Dehaene,2 Aubree L Hawley,3 Hexirui Wu,3Elisabet Borsheim,4 and John Van Camp2

1University of Arkansas, Department of Food Science, Centerfor Human Nutrition; 2University of Gent, Belgium; 3University ofArkansas; and 4University of Arkansas for Medical Sciences

Objectives Overweight and obesity continue to be public healthissues. Identifying novel nutrition strategies is important for thetreatment and prevention of weight gain. The objective of this studywas to determine the effect of consuming omega-3 fatty acids (O3FA)on postprandial appetite, energy expenditure, and substrate oxidationin normal weight (NW) and overweight or obese (OW) young women.

Methods: In total, 29 young normal weight [NW; n = 15; age:24.1 ± 3.0 y; body mass index (BMI): 21.2 ± 1.6 kg/m2] and OW(n = 14; age: 24.7 ± 3.8 y; BMI 28.9 ± 3.9 kg/m2) women wererecruited to participate in this randomized, double-blind, crossoverstudy. Participants consumed a control and anO3FA test beverage, withawashout period of≥1wk between studies. The test beverage contained5 g of O3FA (2 g of eicosapentaenoic acid; 2 g of docosahexaenoicacid) and the control beverage contained 5 g of corn oil. Participantsarrived fasted. Their energy expenditure (EE) and substrate oxidationwere measured using indirect calorimetry and their appetite wasmeasured using visual analog scales over a 4-h period. Their foodintake was measured using weighed food records over the remaining24-h period. Data were analyzed using 2-factor, crossover, repeated-measures ANOVA or 2-sample independent t test.


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Results: O3FA consumption increased postprandial niAUC EE inNWparticipants and inOWparticipants when controlled for leanmass.There were effects of diet (P < 0.0001) and time (P < 0.0001) and asignificant interaction of diet and time (P< 0.0001) on EE. Postprandialfat oxidation was increased with O3FA (P = 0.05) in both NW andOWparticipants. BothNWandOWparticipants had decreased hunger(P < 0.01), desire to eat (P < 0.01), and prospective food consumption(P < 0.001) with increased feelings of fullness (P < 0.01) over the 4-hpostprandial period with O3FA compared with control. There was nosignificant effect of O3FA intake at breakfast on food intake later in theday in NW or OW participants.

Conclusions: This study is one of the first to demonstrate thatconsumption of O3FA at breakfast has the potential to increasepostprandial EE and fat oxidation and to decrease hunger in both NWand OW young women. Longer-term interventions are needed.

This trial was registered at clinicaltrials.gov as NCT03079635.Funding SourcesArkansas Biosciences Institute. The beverages were supplied by

Smartfish (Oslo, Norway).

Relationships between Energy Expenditure and RespiratoryQuotient in Obese Adults after a Moderate 8-wk Weight LossIntervention (P10-008)

Jacqueline A Beatty and Kathleen Melanson

The University of Rhode Island

Objectives: Maintaining weight loss is difficult, in part owing tothe changes in resting metabolic rate (RMR) and substrate oxidationthat accompany weight reduction. To date, most research has examinedRMR and substrate oxidation after moderate to large body weightchanges. In reality, many weight loss attempts result in only minorweight changes. This study examined energy expenditure and substrateoxidation, as reflected by respiratory quotient (RQ), within an 8-wk,self-led weight loss intervention.

Methods: In an experimental design, 22 adults (13 females; age:34.6 ± 16.5 y; body mass index: 32.0 ± 4.3 kg/m2) participated in an8-wk weight loss intervention. Body weight, percentage body fat (BodPod), and estimated energy expenditure from a 7-d physical activityrecall (PAR-EE) were collected at weeks 0 and 8. At these time points,indirect calorimetry was also used to measure 45-min fasting RMRand RQ. Paired t tests examined changes over time, and correlationsexplored associations between percentage body fat, PAR-EE, and RQ.

Results: Pre-post changes (mean± SD) in body weight (–0.72± 2.0kg), percentage body fat (–0.77% ± 10.5%), PAR-EE (+180.1 ± 584.0kcal/d), RMR (+14.7 ± 108.8 kcal/d), and RQ (+0.14 ± 0.05) werenot significant. A significant, but moderate, negative correlation wasfound between week 8 PAR-EE (1651.7 ± 568.7 kcal/d) and week 8RQ (0.85 ± 0.04; r = –0.433; P = 0.04). A significant, but moderate,negative association was also found between week 8 percentage bodyfat (36.8% ± 9.2%) and week 8 RQ (0.85 ± 0.04; r = –0.451;P = 0.04).

Conclusions: Participants with increased physical activity after an8-wk weight loss intervention tended to have higher fasting fatoxidation. At 8 wk (but not 0 wk), higher body fat was associated

with higher fasting fat oxidation. Although not significant, changes inbody weight, percentage body fat, and estimated energy expenditurewere associated with increased RMR, in contrast to findings with largerchanges. These data suggest that even with minor changes in physicalactivity and body fat, increased energy expended in physical activitymay be associated with greater fat oxidation, which may favor weightloss maintenance. Prospective research is needed to follow this upover longer periods.

Funding SourcesA grant awarded by the Obesity Society funded this research.

A Pilot Randomized Controlled Trial to Assess the Effects ofa Daily Pistachio (Pistachia vera) Afternoon Snack on Next MealEnergy Intake, Satiety, and Anthropometry in French Women(P10-009)

France Bellisle,1 Anestis Dougkas,2 Agnes Giboreau,2 AriannaCarughi,3 and Jennette Higgs4

1Epidémiologie Nutritionnelle, Université Paris, France; 2InstitutPaul Bocuse, France; 3American Pistachio Growers; and 4Food to Fit

Objectives:Nuts, including pistachios, are rich in fiber and protein,which have been shown to enhance satiety. Studies have shown thatincluding nuts in the diet improves appetite control and does not leadto weight gain. The aim of this study was to investigate the dailyconsumption of pistachios as an afternoon snack in the workplace orat home, and its effects on satiety, energy and nutrient intake, and bodyweight and composition.

Methods: This pilot, monocentric, randomized controlled trialincluded 2 parallel groups of 30 healthy, sedentary women, aged 18–50 y. For 4 wk, groups consumed as an afternoon snack either 56 g(315 kcal) of roasted, lightly salted pistachios or 56 g of commerciallyavailable, isocaloric or equiprotein savoury biscuits. Evening energyintake after the afternoon snack, changes in anthropometric measures,and daily intake of energy, macronutrients, and selected micronutrientswere assessed. Visual analog scales (VAS) were used to rate sensationsof hunger, thirst, fullness, desire to eat, and prospective consumption.

Results: Satiety effectswere not different between groups, as assessedby evening energy intake or VAS scores (P < 0.05). Neither consumingpistachios nor eating biscuits had an impact on body weight after4 wk. This could be attributed to compensationmechanisms. There wasa trend toward smaller waist circumference in the pistachio group. Fatbody mass remained stable among the women consuming pistachios,whereas it increased slightly among those consuming biscuits. Leanbody mass decreased slightly in this group, but did not change amongthe women consuming pistachios. Thiamine, vitamin B-6, copper,and potassium intakes were significantly higher during the pistachiointervention compared with the control condition.

Conclusion: The lack of difference in subjective feelings of hungeror satiety, or in food consumption between the snacks, can be explainedby their similar energy and protein contents. A daily snack of 315 kcalof pistachios for 1 mo did not affect body weight or anthropometricmeasures, but it did significantly improve the intake of micronutrients.

Funding SourcesAmerican Pistachio Growers.


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Integrative Network Analysis of Body Composition, PlasmaRedox, Dietary Intake, and Plasma High-Resolution Metabolomics(P10-010)

Moriah P Bellissimo, Karan Uppal, Phong H Tran, Erin CMillson, Dean P Jones, Thomas Ziegler, and Jessica A Alvarez

Emory University, GA

Objective: Adiposity, oxidative stress, and diet have known effectson health and disease; however, their combined effect on metabolismis unknown. We aimed to examine the interrelations of body com-position, aminothiol redox profile, and dietary intake with metabolicpathways derived from plasma high-resolution metabolomics (HRM)with the use of newly developed software enabling integrative dataanalysis.

Methods: Subjects included 180 working adults (64% female;mean ± SD age: 49.5 ± 10.3 y) in an Atlanta cohort. xMWASwas used for integrative network analysis followed by communitydetection to identify communities and clusters of highly connectednodes by incorporating dual-energy X-ray absorptiometry (21 vari-ables), plasma aminothiol redox (10 variables), Block food frequencyquestionnaire (337 variables), and HRM (10,211 metabolic features)data. Pathway analysis of metabolic features in select communities wasperformed using Mummichog. All results reported have a P-value of<0.05.

FIGURE 1 Integrative network analysis.

Results: Integrative network analysis showed plasma glutathione(GSH) was inversely associated with visceral adipose tissue (VAT;r = –0.72) and lean body mass (LBM; r = –0.83), and positivelyassociated with gynoid fat (r = 0.61). Plasma cystine (CySS) and theredox ratio (CySS:GSH) were positively associated with percentagebody fat (r = 0.93, r = 0.75, respectively), total fat (r = 0.83, r = 0.92),subcutaneous fat (r= 0.91, r= 0.84), and android fat (r= 0.86, r= 0.75).In addition, xMWAS identified 5 unique communities (C1–C5). C1

was comprised of CySS:GSH, percentage body fat, percentage LBM,and vitamin D and fatty acid metabolites. C2 included VAT measuresand amino acid (AA) metabolism pathways. C3 included androidfat, and pathways involved in linoleic acid and glycerophospholipidmetabolism. C4 included subcutaneous fat, total body fat, and totalbodyweight, withAA and butanoatemetabolismpathways. C5 includedmicronutrients, fiber, and essential fatty acids, with markers of bone,lean and fat mass, and aminothiol redox.

Conclusions: Plasma GSH was associated with more favorable fatdistribution, whereas a more oxidized plasma aminothiol profile waspositively associated with general adiposity. Using a novel approach,integrative network and community detection analyses identified com-munities of individual components of body composition, aminothiolredox, and diet linked to specific metabolite signatures and pathways.

Funding SourcesThis study was supported by the NIH.


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Acute Effect of an Egg-Based High-Protein Meal on the Hyper-tensive and Endothelial Responses to Exercise (P10-011)

Robert E Bergia,1 Wayne Campbell,1 Bruno T Roseguini,1 andJung E Kim2

1Purdue University, IN; and 2National University of Singapore

Objectives: Exercise training ultimately improves health, but acuteexercise triggers an increase in blood pressure (BP), which may bedisadvantageous to health. Chronic ingestion of elevated dietary proteincan reduce blood pressure. Therefore, consumption of a higher-proteinmeal may attenuate the acute hypertensive response to exercise andallow individuals at risk for hypertension to experience the healthbenefits of both acute exercise and training. We hypothesized that theconsumption of a meal with 30 g of protein (HP; 23 g from egg sources)would attenuate exercise-induced systolic BP elevation and improve BP-related markers of endothelial function, compared with an isocaloricmeal containing 13 g of protein (LP; 7 g from egg sources).

Methods: In total, 31 subjects (18 male, 13 female; age: 33 ±14 y; body mass index: 26.5 ± 3.8 kg/m2; systolic BP: 127 ± 5 mmHg; diastolic BP: 77 ± 6 mm Hg) completed this randomized, double-blind, crossover acute feeding study. On each testing day, subjectsconsumed either an HP or LP meal and 165 min later, exercised ona cycle ergometer at 70% maximal oxygen uptake for 30 min. BPwas measured immediately prior to the meal (0 min) and periodicallybefore, during, and after exercise over a 315-min period. Concurrentwith BP measurements, blood was sampled to determine endothelialfunction via quantification of plasma nitrites, nitrates, endothelin-1,arginine, and arginine metabolites.

Results: Consuming an HP meal did not attenuate BP responses toexercise. There was no differential response in the 30-min compositesystolic BP incremental area under the curve (iAUC) (least-squaresmean ± SE: HP: 464 ± 37, LP: 457 ± 37 mm Hg; P = 0.76) or peaksystolic BP (HP: 190± 5, LP: 190± 5mmHg; P= 0.90) during exerciseby meal. Time for systolic BP to return to baseline postexercise BP(3 min recovery ratio; HP: 0.80 ± 0.02, LP: 0.78 ± 0.02; P = 0.49)and postexercise systolic BP iAUC (HP: 102 ± 15, LP: 67 ± 15 mmHg; P = 0.07) were not influenced by protein intake. There was nodifferential response in any plasma marker of endothelial functionduring exercise by meal.

Conclusion: These results suggest that the chronic beneficial effectsof consuming greater dietary protein on BP do not manifest in acutesettings in which participants consume higher protein meals and thecardiovascular system is challenged with aerobic exercise.

Funding SourcesFunded by the American Egg Board-Egg Nutrition Center.

Effect of Whey Protein Supplementation on Body Composi-tion Changes in Women: A Systematic Review and Meta-analysis(P10-012)

Robert E Bergia, Joshua Hudson, and Wayne W Campbell


Objectives: Whey protein (WP) supplementation is promotedto help women improve their body composition, especially whenconsumed in conjunctionwith energy restriction (ER) and/or resistancetraining (RT). However, a perception exists that women should not

consume WP supplements because it could make them “bulky”(i.e., excessive lean mass accretion, which includes skeletal muscle).Although results from individual research studies do not support thenotion that WP supplementation makes women “bulky,” a systematicassessment of the issue is needed. The purpose of this research was toassess the effect of WP supplementation on body composition changesover time in adult women.

Methods: A systematic review and meta-analysis of literature wasconducted to specifically determine the effect of WP supplementationin a 2 × 2 factorial fashion: with or without ER, and with orwithout RT. For this, Pubmed, Scopus, Cochrane, and CINAHL weresystematically searched. Two researchers independently screened 1845abstracts and extracted 276 articles. In total, 13 randomized controlledtrials (resulting in 15 comparisons) with 28 groups met all inclusioncriteria.

Results:Globally,WP supplementation increased leanmass (WMD:0.37 kg; 95% CI: 0.06, 0.67) while not influencing changes in fat mass(–0.20 kg; 95% CI: –0.67, 0.27) relative to the non-WP control. Thebeneficial effect of WP on lean mass was lost when only includingstudies with RT in the analysis (n = 7 comparisons; WMD: 0.23 kg;95% CI: –0.17, 0.63). The beneficial effect of WP on lean mass wasmore robust when only including studies with an ER component (n= 6comparisons; WMD: 0.90 kg; 95% CI: 0.31, 1.49). There was no effectof WP on lean mass in studies without ER (n = 9 comparisons; 0.22 kg;95% CI: –0.12, 0.57).

Conclusion: Overall, WP supplementation equated to ∼1% greaterlean mass in women over the course of these trials. Considering thatWP supplementation did not influence fat mass, this 1% differencein lean mass does not support the public perception that WP makeswomen “bulky.” Althoughmore research is needed to specifically assessthe effects in varying states of energy sufficiency and exercise training,the overall findings support consumption of WP in women seeking tomodestly improve body composition.

Funding SourcesWhey Protein Research Consortium.

Dietary Patterns Have Distinct Metabolomic Profiles amongAsian Indian Adults in theMetabolic Syndrome and Atherosclerosisin South Asians Living in America (MASALA) Study (P10-013)

Shilpa N Bhupathiraju,1 Marta Guasch-Ferre,2 Meghana DGadgil,3 Christopher B Newgard,4 James Bain,4 MichaelMuehlbauer,4 Olga Ilkayeva,4 Denise M Scholtens,5 Frank BHu,2 Alka M Kanaya,6 and Namratha Kandula7

1Channing Division of NetworkMedicine, HarvardMedical School,MA; 2Harvard TH Chan School of Public Health, MA; 3Universityof California, San Francisco; 4Duke University Medical Center, NC;5Northwestern University, IL; 6University of California, San Francisco;and 7Northwestern University Feinberg School of Medicine, IL

Objectives: Different dietary patterns have distinct metabolomicsignatures that may influence disease risk. However, evidence fromSouth Asians, a group with unique dietary patterns and a highprevalence of cardiometabolic risk, is lacking.

Methods: We investigated the metabolomic profiles associatedwith 2 distinct dietary patterns among Asian Indians living in theUnited States. We also examined cross-sectional associations between


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metabolomic profiles and cardiometabolic riskmarkers.We used cross-sectional data from 150 Asian Indians, aged 45–79 y, in the MetabolicSyndrome and Atherosclerosis in South Asians Living in America(MASALA) pilot study. Metabolites were measured in fasting serumsamples. Subjects’ usual diet was assessed using a validated foodfrequency questionnaire. We used principal components analysis toderive dietary and metabolomic patterns. We used adjusted generallinear regression models to examine associations between dietarypatterns, metabolite patterns, and cardiometabolic risk markers.

Results: We observed 2 major metabolomic patterns—a medium-to long-chain acylcarnitines pattern (metabolite pattern 1) and abranched-chain amino acids, aromatic amino acids, and short-chainacylcarnitines pattern (metabolite pattern 2). In both the overallpopulation and in a subsample of participants without type 2 diabetes(n = 105), a “Western/nonvegetarian” diet pattern was significantlyand positively associated with metabolite pattern 2 (Table 1). Highermetabolomic pattern 2 scores were adversely associated with measuresof glycemia (including measures of fasting glucose, insulin, 2-h insulin,

insulin sensitivity index, and insulin resistance), total adiponectin,lipid measures (including total cholesterol and triglycerides), and aradiographic measure of hepatic fat (liver-spleen attenuation).

Conclusions: Our findings suggest that a “Western/nonvegetarian”dietary pattern is associated with a metabolomic profile linked withan adverse cardiometabolic profile. Public health efforts to reducecardiometabolic disease burden should focus on consuming a healthyplant-based diet.

Funding SourcesThe MASALA pilot study was funded by grant no. K23 HL080026

from the University of California, San Francisco, Research Evaluationand Allocation Committee, and by NIH/National Center for ResearchResources University of California San Francisco-Clinical and Transla-tional Science Institute grant no. UL1 RR024131. SNB is supported by acareer development award from the National Institute of Diabetes andDigestive and Kidney Diseases (NIDDK), NIH (K01 DK107804). JRBwas supported by NIDDK (P01 DK058398) and the National Instituteon Aging (P30 AG028716).


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Just Another Bite! An Analysis of Bites’ Caloric Variability in aControlled Dietary Setting (P10-014)

Anna Bolzon,1 Elisa Fuscà,1 Alessia Buratin,1 AlessandraCorazzina,1 Lorenza Caregaro,2 Romina Valentini,3 IleanaBaldi,1 Egle Perissinotto,1 and Dario Gregori1

Departments of 1Cardiac, Thoracic and Vascular Sciences and2Medicine (DIMED), University of Padova, Italy; and 3Hospital ofPadova, Italy

Objective: Caloric intake is known to be a pillar in dietaryintervention, but very little information is available on its variabilityin relation to the main actions associated with eating, like bites.Nevertheless, bites are known to be associated with major eatingparameters, like speed and per-unit amount of food intake. The aim ofthis studywas to investigate the variability in the energy content of bites.

Methods: A total of 13 types of packaged foods were selectedand submitted to 30 volunteers (14 males and 12 females) in a fullycontrolled dietary feeding intervention. For each food item, 10 biteswere collected, and 2 bites of each group were randomly selected forthe measurement of the energetic content by bomb calorimetry IKAC200. The corresponding metabolizable energy values were calculatedby subtracting 1.25 kcal/g protein from the gross energy to correct forincomplete digestibility. As a secondary outcome, the actual bite caloriccontent was compared with the expected caloric intake as specifiedon the food labels. The statistical analysis was based on descriptivemeasures with 95% CIs derived via bootstrap resampling.

Results: Variability in the caloric intake of bites, expressed in termsof CV%, was high, in particular for chicken meatballs with tomatosauce (CV%: 13.4%; 95%CI: 12.5%, 0. 15%), tagliolini with mushrooms(CV%: 16.6%; 95% CI: 15.5%, 18.4%), vegetable soup (CV%: 20.5%;95% CI: 17.5%, 24.8%), chicken and artichokes (CV%: 41.2% (34.6%,48.7%), and eggplant parmigiana (CV%: 27.8%; 95% CI: 24.6%, 32.4%).The highest percentage of mean differences between the calories of thebites and the respected stated value were detected using the stated valueas the reference. The main results were CV%: 41% (95% CI: 35.4%,46.5%) for chicken meatballs with tomatoes; CV%: –20.7% (95% CI:–30.2%, –10.7%) for chicken and artichoke; CV%: 18.6% (95%CI: 8.6%,28.2%) for eggplant parmigiana; CV%: 14.8% (95% CI: 9.1%, 22.6%)for sandwiches; CV%: 6.7% (95% CI: 3.6%, 9.6%) for mozzarella; andCV%: 7.5% (95% CI: 5.7%, 9.2%) for Italian fresh cheese.

Conclusions:Variability in caloric intake of bites is noteworthy, andit seems to be higher in foods with a complex matrix or with a highintrinsic humidity. Remarkably, the data show an overall tendency ofcalorimetry to overestimate the caloric content in comparison with thelabeled caloric value.

The Potential for Omega-3 Fatty Acids to Increase EnergyExpenditure in Normal Weight and Overweight or Obese Children(P10-015)

Elisabet Borsheim,1,2,3 Matthew Cotter,3 Charlayne F Mitchell,4Sarah L Russell,4 Hexirui Wu,4 Aubree Hawley,4 Eva Dehaene,5and Jamie Baum4

1Arkansas Children’s Nutrition Center; 2Arkansas Children’sResearch Institute; 3University of Arkansas for Medical Sciences;4University of Arkansas; and 5University of Gent, Belgium

Objectives: The objective of this study was to determine if2 wk of omega-3 fatty acid (O3FA) supplementation increaseswhole-body energy expenditure (EE) and improves cardiometabolichealth markers in normal weight (NW) and overweight or obese(OW) children.

Methods: A total of 20 male (M) and female (F), NW [n = 11(M = 5, F = 6); age: 9.8 ± 0.4 y; body mass index (BMI) percentilefor age and sex: 49.5% ± 6.9%] and OW [n = 9 (M = 5, F = 4);age: 10.2 ± 0.5 y; BMI percentile: 91.1% ± 1.5%) children wererecruited to participate in this randomized, crossover, double-blindstudy. Participants consumed isocaloric (320 kcal), macronutrient-matched beverages every morning before 1000 for 14 d, with a washoutperiod of ≥14 d between interventions. The O3FA beverages weresupplemented with 5 g of O3FA (2 g of eicosapentaenoic acid; 2 gof docosahexaenoic acid), whereas the control beverages contained5 g of corn oil. Resting EE (REE) and markers of cardiometabolichealth were measured at baseline (day 0) and day 14. Total EE(TEE) was measured using doubly labeled water. Food intake wasmeasured during each week of the intervention using 3-d weighedfood records. Data were analyzed using 2-way ANOVA or 2-sampleindependent t test.

Results: There was no effect of O3FA on REE (day 14 vs. day 0, NS)in either NW or OW participants when controlling for fat-free mass(FFM). Breakfast intake for 14 d tended to decrease fat oxidation (day0: 0.009 ± 0.001 vs. day 14: 0.004 ± 0.002 kcal · min–1 · kg–1 FFM)and increase carbohydrate oxidation (day 0: 0.021 ± 0.002 vs. day 14:0.026 ± 0.002 kcal · min–1 · kg–1 FFM; P = 0.09) in OW children only,with no effect of O3FA. Although not significant, TEE was 130 kcal/dhigher in NW following O3FA supplementation for 14 d comparedwith control drink (2060 ± 276 vs. 1930 ± 176 kcal/d, respectively)and 75 kcal/d higher in OW following O3FA supplementation for14 d compared with control drink (2469 ± 221 vs. 2393 ± 268kcal/d). There was no effect of O3FA supplementation on markers ofcardiometabolic health (fasting glucose, free fatty acids, triglycerides,leptin, and adiponectin concentrations).

Conclusions: There was no significant effect of 14-d O3FA supple-mentation on EE or markers of cardiometabolic health in NW or OWchildren. However, there were indications that O3FA supplementationmay impact total energy expenditure over time. Future research requiresa larger sample size and longer intervention time.

This trial was registered at clinicaltrials.gov as NCT03139773.Funding SourcesArkansas Biosciences Institute. Beverages were provided by Smart-

fish (Oslo, Norway).

In vitro Study of Micellar Casein and Caseinate Digestion (P10-016)

Audrey Boulier,1 Alain Baniel,1 Sylvain Denis,2 and MoniqueAlric2

1Ingredia SA; and 2Clermont Auvergne University, France

Objectives: The digestion of soluble milk proteins (SMPs), so-called“fast” proteins, is different thanmicellar caseins (MCs), so-called “slow”proteins. Unlike SMPs, MCs clot in the stomach at an acidic pH,resulting in decreased gastric emptying time (GET) and a sustainedrelease of amino acids into the blood. There is limited information on


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the link between structure anddigestion of caseins. The aimof this studywas to study the impact of the denaturation of caseins during digestionunder in vitro conditions.

Methods: The products [MCs, sodium caseinate (NC)] were di-gested in the TIM system, a dynamic tool mimicking human digestion.The proteins were digested alone (n = 3), according to a protocolsimulating a meal’s digestion in a healthy adult. Tests measuringnitrogen balance, the release of total nitrogenous matter, and peptideprofiles (PPs) were performed after 5 h of digestion. For statisticalanalysis, we used the Kruskal-Wallis test.

Results: In the stomach, the caseins rapidly precipitated, but withdifferent coagulum, more or less compact, which led to a decrease inGET. The GET was significantly faster (P < 0.01) with the NC thanwith the MCs. The PPs of the liquid fractions were also different forthe 2 substrates. For the MCs, more molecules >10 kDa remainedintact at the beginning of digestion (61% at 50 min) but decreased withtime (13% at 5 h). For NC, the pattern was reversed (fractions >10kDa= 11% at 50min and 38% at 5 h). These results follow the solubilityof its coagulum. In the small intestine, the digestibility of the testedproteins was very high (>90%). The majority of the absorption waslocated in the jejunum.

Conclusion: The change in structure modifies the digestive processof proteins and thus the protein metabolism.

Funding SourcesIngredia SA.

Associations between DASH Diet Score and Measures ofGlycemia and Insulin Action in Obese Adolescents (P10-017)

Ethan M Braun,1 Kendra Wilder,1 Lisa Smith,2 Carol Boushey,3Edward Delp,1 Tamara Hannon,2 and Nana Gletsu-Miller1

1Purdue University, IN; 2Indiana University; and 3University ofHawaii Cancer Center

Objective: The Dietary Approaches to Stop Hypertension (DASH)diet is a healthful dietary pattern that may reduce the risk of developingtype 2 diabetes (T2D) in adolescents.We aimed to determine the impactof adherence to the DASH diet on measures of glycemia and insulinaction in obese adolescents.

Methods: Baseline measures were obtained from an ongoing, ran-domized, controlled intervention to prevent T2D in obese adolescents.The Technology Assisted Dietary Assessment system (TADA) was usedto create a 4-d mobile food record. Glucose and insulin dynamics,including fasting and 2-h glucose concentrations, homeostatic modelassessment insulin resistance index (HOMA-IR), whole-body insulinsensitivity index (WBISI), and dispositional index (DI) were assessedduring a 2- h oral glucose tolerance test (OGTT). DASH scoreswere calculated for each participant and separated into quintiles.Independent sample t tests assessed differences between 1st and 5thDASH score quintiles and biomarkers from the OGTT. Values areexpressed as mean ± SD.

Results:Obese adolescents (n= 22, comprising 9 boys and 13 girls),were screened and 50% were found to have prediabetes. The bodymass index of the participants was 34.6 kg/m2. Glycated hemoglobinconcentrations were lower for participants who hadDASH scores in the5th quintile versus the 1st quintile (5.12± 0.17 vs. 5.42± 0.16;P< 0.05).A tendency for more favorable biomarkers was observed for fasting and

2-h glucose concentrations, HOMA-IR, WBISI, and DI, but none werestatistically significant.

Conclusions: Adherence to the DASH dietary pattern may improveglycemia in adolescents. Prospective studies are needed to investigatethis question. TADA is a useful tool for assessment of dietary intake inadolescents.

Funding SourcesFunded by a RalphW and Grace M Showalter Research Trust grant,

Purdue University.

Role of the Intestine and Its Contents in Formate Production inthe Rat (P10-018)

John T Brosnan, Kathy Clow, Theerawat Pongnopparat, andMargaret Brosnan

Memorial University of Newfoundland, Canada

Objectives: Pectins are polysaccharides that are rich in galacturonicacid, of which ∼80% of the carboxyl groups are esterified withmethanol. These methyl groups may be removed by methylesterasesduring digestion.We hypothesized thatmethanol so producedmay giverise to substantial quantities of formate, which becomes available to rattissues as a source of one-carbon groups. The aim of this study was totest that hypothesis.

Methods: Formate and methanol were measured along the gas-trointestinal (GI) tract of chow-fed Sprague-Dawley rats, as well as inthe hepatic portal vein (HPV) and the hepatic vein, and in an artery.A separate group of chow-fed rats were treated with oral antibiotics(vancomycin, gentamycin, rifampin) for 7 d. We also examined ratsfed AIN-based diets containing either 5% pectin (PF) or 5% cellulose(CF) as their fiber source. Formate was analyzed by GC-MS andmethanol was analyzed by headspace GC-MS. Blood flow in the HPVwas determined via a flow probe.

Results: Formate was found throughout the GI tract of chow-fedrats, but was most abundant in the ileum (ileum > cecum > largeintestine). Confirming the important role of the small intestine inthe provision of formate to the liver, we found that the formateconcentration in the superior mesenteric vein (largely drains the smallintestine) was appreciably higher than that in the inferior mesentericvein (largely drains the large intestine). Formate was particularly foundin the intestinal contents rather than intestinal tissue. Antibiotic-treated, chow-fed rats had decreased plasma formate concentrationsand a complete suppression of intestinal formate output. The intestinalformate content wasmuch higher in PF rats than in CF rats; liver uptakeof methanol was also appreciably higher in the PF rats. Circulatingmethanol concentrations in the PF rats (800 µM) were >100-foldgreater than in the CF rats.

Conclusions: The intestinal microbiome and the source of dietaryfiber interact in determining methanol and formate production by theGI tract. Methanol, produced from pectin, gives rise to substantialquantities of formate, which become available to rat tissues as a potentialsource of one-carbon groups. These studies are also relevant to studieswith vitamin B-12–deficient animals, in which pectin is commonly usedas a cobalamin-binding agent.

Funding SourcesCanadian Institutes for Health Research.


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Protein Supplementation, PlasmaBCAAs, and InsulinResistancein Postmenopausal Women: An Ancillary Study from a WheyProtein Supplementation Trial (P10-019)

Alessandra Byer,1 Jessica Bihuniak,1 Christine Simpson,2 Karl LInsogna,2 Jeannette M Beasley3

1New York University; 2Yale University School of Medicine, CT; and3NYU School of Medicine

Objective: Recent studies have reported an increased risk of type 2diabetes among people with higher protein intake.Moreover, branched-chain amino acids (BCAAs) are reported to be positively associatedwithinsulin resistance (IR). However, it is not understood whether elevatedconcentrations of BCAAs are causal to IR development, or if higherBCAAs are a marker of IR. The objective of this study was to examinethe effects of long-term protein and carbohydrate supplementation onplasma BCAA concentrations, and the relation between plasma BCAAsand IR in older women.

Methods: Stored samples and data from 84 postmenopausal womenwho had previously participated in a protein supplementation trialwere included in this ancillary study. Subjects had been randomizedto a whey protein supplement (PRO; n = 38) or a maltodextrinsupplement (CHO; n = 46) for 18 mo. Plasma BCAAs, homeostaticmodel assessment (an IR assessment method), and body compositionwere analyzed at baseline and 18 mo. Descriptive statistics were usedto summarize participant characteristics, BCAA concentrations, andoutcome measures. Changes in plasma BCAA concentrations, IR, andbody composition were compared by study arm using Mann-WhitneyU tests. Associations between BCAA concentrations, IR, and bodycomposition were assessed using Spearman correlations by study armand time point.

Results: There were no significant changes in plasma BCAAconcentrations or IR by study arm. At baseline, there were no significantassociations between plasma BCAA concentrations and IR (CHO:r = 0.269, P = 0.071; PRO: r = 0.270, P = 0.101). However, there was asignificant positive association between plasma BCAA concentrationsand IR in both groups at 18 mo (CHO: r = 0.353, P = 0.016; PRO:r = 0.352, P = 0.030). Total lean mass decreased in the CHO group(total: –0.6 kg) but increased in the PRO group (total: 0.2 kg, P= 0.033).The nonsignificant within-group changes in fat mass did not differbetween treatment groups.

Conclusions: Plasma BCAA concentration did not significantlychange after 18 mo of supplementation, but is positively correlated withIR after 18 mo of protein and carbohydrate supplementation. Futurework will examine whether correlations are because of changes in bodycomposition, physical activity, and/or energy intake.

Funding SourcesThis project was supported in part by a grant from the New York

University Research Challenge Fund Program.

Muscle Proteolysis Is Largely Unaffected by Severe EnergyDeficitduring High-Altitude Acclimatization (P10-020)

JohnWCarbone,1 LeeMargolis,2 Claire Berryman,2 ChristopherCarrigan,2 Nancy Murphy,2 Arny Ferrnado,3 Andrew J Young,2and Stefan M Pasiakos2

1Eastern Michigan University; 2US Army Research Institute ofEnvironmental Medicine; and 3University of Arkansas for MedicalSciences

Background: At sea level (SL), sustained energy deficit (ED) elicitsfat-free mass (FFM) loss without significant changes in ubiquitin-mediated proteolysis (UMP). Chronic exposure to high altitude (HA)elicits muscle atrophy in lowlanders, although past animal and humanstudies have provided conflicting results regarding the influence of HAand the associated exercise- and diet-induced ED on UMP at rest andpostexercise. Additionally, although consuming high-quality proteinattenuates UMP at SL, this effect has not been studied at HA.

Objective: The aim of this study was to examine the effectsof HA and concomitant ED on basal, postexercise, and recoverymuscle proteolytic events in recreationally active lowlanders (n = 8men) who completed sequential 21-d dietary and environmentalphases: 1) eucaloric feeding at SL; and 2) exercise- and diet-inducedED (–1766 ± 145 kcal/d) at HA.

Methods: Body composition was assessed by dual-energy X-ray absorptiometry at SL and immediately after descent from HA.Vastus lateralis muscle samples were obtained at SL, acute HA, andchronic HA (21 d of ED) before (BASE), after 80 min of steady-state exercise and ensuing 2-mile time trial (POST), and 3-h intorecovery after consuming 25 g of whey protein postexercise (REC).Expression and activity of UMP factors were assessed using quantitativereverse transcriptase-polymerase chain reaction, Western blotting, andenzymatic activity assays.

Results: Mean ± SEM total weight and FFM loss over 21 d of EDat HA were 7.7 ± 0.8 and 4.3 ± 1.0 kg, respectively. Basal expressionand activities of all proteolytic measures were not different across thephases. Independent of phase, p-FOXO1Thr24 was lower at REC than atBASE and POST (P < 0.05). MURF1 was higher at REC than at BASEand POST (P 0.05), and lower at chronic HA than at SL (P < 0.05).Independent of phase, ATROGIN-1 was lower at POST than at BASE,and was further reduced at REC (P < 0.05). Protein ubiquitylation washigher at POST than at BASE and REC at SL, and POST ubiquitylationwas lower at acute and chronic HA than at SL (P < 0.05; phase-by-timeinteraction). Independent of phase, β1 and β526S proteasome activitieswere lower at POST than at BASE and REC (P < 0.05).

Conclusions: These data show that the stress of severe ED duringHA acclimatization did not upregulate markers of muscle proteolysis atrest or after exercise and recovery protein consumption, suggesting thatthe role of UMP in HA-mediated muscle atrophy is likely to be trivial.

Funding SourcesSupported by US Army Medical Research and Materiel Command.

Modulation of Ghrelin Homeostasis after Oral Ingestion ofMacronutrients in Healthy Men (P10-021)

Maritza E Carrillo,1 Johannes Veldhuis,2 and Ali Iranmanesh3

1Department of Veterans Affairs & Carilion Clinic Virginia TechSchool of Medicine; 2Mayo Clinic, MN; and 3VA Medical Center

Objectives: This study was designed to assess nutrient-inducedvariation in circulating ghrelin concentrations after oral ingestion ofthe 3 macronutrients per se, and as it relates to measures of glucosehomeostasis and satiety.


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Methods:Agroup of 11 healthymen (age: 33–70 y; bodymass index:20.4–31.5 kg/m2) were studied after overnight fast on 4 separate days,involving oral ingestion of 300 ml of either water, dextrose, protein,or lipid solution. Test meals were isocaloric (400 kcal). Sessions were6.5 h long, starting at 0800–0900. Blood was collected at 10-minintervals for ghrelin (pg/mL), glucose (mg/dL), insulin (µIU/mL), andC-peptide (ng/mL) measurements. Satiety scale was applied at 10-minintervals. ANOVA and multiple regression were used for data analysis.

Results: In contrast to a gradual increase from the fasted state (con-trol), circulating ghrelin concentrations were decreased to comparablelevels (P = 0.09) after oral ingestion of macronutrients, but with asignificantly shorter time lag (P = 0.0006) for dextrose (101 ± 13min) than for protein (174 ± 10 min) and lipid (185 ± 20 min).The postingestion ghrelin nadir levels rebounded to concentrationshigher than baseline, without any significant difference between the 3macronutrients (P= 0.36). Recovery time to baseline values was longerfor protein (330± 17min; r2 = 0.77), and after dextrose (r2 = 0.83) andprotein (r2 = 0.53) intake, but glucose after lipid ingestion (r2 = 0.14);and (2) ghrelin as the major determinant of satiety state only afterdextrose (r2 = 0.64), as opposed to C-peptide during fasting (r2 = 0.86),and after protein (r2 = 0.29) and lipid (r2 = 0.95) meals.

Conclusion: Suppression of circulating ghrelin concentrations andthe corresponding rebound to values above baseline were observedwith all 3 macronutrients. Although the nadir and peak levels werecomparable, times to nadir and recovery to baseline were significantlyshorter after dextrose ingestion. The longer duration of ghrelinsuppression after protein and lipid meals could be interpreted as a moreprolonged satiety period, which warrants further investigation.

Funding SourcesDepartment of Veterans Affairs.

Identifying Differences in Metabolite Patterns Owing to HighFat versus High Carbohydrate Diet Using Metabolomics in Humans(P10-022)

James L Casey

University of Michigan

Objectives: Nutritional research frequently involves the use ofself-reported measures of food intake such as dietary recall andfood frequency questionnaires, which may be subject to participants’systematic errors. The objective of this studywas to explore and examinethe use of metabolomics as a less biased form of diet identification tool.The hypothesis explored by this study was that metabolite profiles insubjects will show specific and identifiable patterns between high-fat(HF) and high-carbohydrate (HC) diets. The objective of this study isto produce a concrete step toward using metabolomics as a form ofunbiased dietary recall.

Methods: In total, 24 metabolically healthy, lean subjects (12 male,12 female) between the ages of 19 and 45 y were recruited for thisstudy. The subjects were weight stable and provided eucaloric dietsthrough the study. A standard diet consisting of prepared foods with15% protein, 35% fat, and 50% carbohydrate was consumed, followedby either an HF or HC diet for 21 d. Plasma was analyzed using LC-MSin a targeted manner.

Results: The variability of 68 targeted metabolites decreased frombaseline after eating a standard diet, as evaluated using Levene’s F test.

This variability increased after eating an HF or HC diet for2 d, and then stabilized between 7 and 14 d. When looking at specificgroups of metabolites, medium-chain acylcarnitines (C6–C16:1) andsuccinate weremost widely differentiated between theHC andHF diets,as identified by t test (p There was a clear convergence of blood plasmametabolites after 3 d of a standard diet. There was also a divergence ofsubjects in the first week of an experimental diet, which then stabilizessomewhere between 7 and 14 d. This shows that the diet has a profoundand measurable effect on plasma metabolites, and indicates that thechanges in plasma metabolites is fairly quick, stabilizing after 1 wk.

Conclusions: It was found that HF diet subjects had elevated con-centrations of medium-chain acylcarnitines and lower concentrationsof succinate and tyrosine after 2 and 7 d. The elevations in acylcarnitineswould seem to indicate that an HF diet involves an increase in theseshort- and medium-chain fatty acids entering the mitochondria. Inconclusion, it appears that there may be an identifiable metabolitepattern that indicates a difference between a high-fat and a high-carbohydrate diet.

Funding SourcesThis research was internally funded from the laboratory of Charles

Burant, University of Michigan.

Omega-3 Polyunsaturated Fatty Acids and CardiometabolicHealth in Chinese Adults: A Prospective Study (P10-023)

Yuming Chen, Gengdong Chen, Ding Ding, Zhanyong Chen,Mianli Xiao, and Jieshen Lin

Sun Yat-sen University, China

Objective: We examined the cross-sectional and prospective asso-ciations of erythrocyte ω-3 (n–3) polyunsaturated fatty acids (PUFAs)with cardiometabolic factors in Chinese adults with habitual diets.

Methods: This community-based prospective study included 3976men andwomen (68%) (age: 40–7 y) inGuangzhou, China. Erythrocyten–3 PUFAs were determined using GC at baseline. The indices ofmetabolic syndrome (MS), carotid artery thickness, and covariates wereassessed at baseline and at the first (+3 y) and second (+6 y) follow-ups.The ultrasound method was used to evaluate nonalcoholic fatty liverdisease (NAFLD) status at the first and second follow-ups. In total, 3075subjects completed the first follow-up and 2660 subjects completed thesecond follow-up and relevant measures.

Results: In the cross-sectional analyses, greater concentrations oferythrocyte n–3 PUFAs were associated with lower prevalences of MS,carotid artery thickening, and NAFLD. The multivariable adjusted ORsfor the comparisons of the extreme quartiles (Q4 vs. Q1) of total n–3 PUFAs were 0.52 (95% CI: 0.32, 0.84) (men) and 0.71 (95% CI:0.53, 0.96) (women) for MS, 0.52 (95% CI: 0.30, 0.90) for carotidartery thickening, and 0.74 (95% CI: 0.61, 0.91) for NAFLD. In theprospective analyses, the multivariable adjusted HRs of the incidenceof cardiometabolic conditions for the extreme quartiles (Q4 vs. Q1) oferythrocyte n–3 PUFAs were 0.74 (95% CI: 0.61, 0.89) for MS, 0.76(95% CI: 0.63,0.91) for hypertension, 0.73 (95% CI: 0.55,0.97) and0.70 (95% CI: 0.55,0.90) for carotid artery thickening at the commoncarotid artery and carotid bifurcation, respectively, and 1.18 (95% CI:1.09, 1.33) for alleviated NAFLD. Pathway analysis showed that thefavorable associations might be mediated by improving inflammatoryfactors (high-sensitivity C-reactive protein, retinol binding protein 4).


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The individual n–3PUFAs (e.g.,α-linolenic acid, eicosapentaenoic acid,docosahexaenoic acid, and docosapentaenoic acid) generally showedsimilar trends to the total n–3 PUFAs.

Conclusions: Our findings show that higher concentrations oferythrocyte n–3 PUFAs were associated with lower prevalence and inci-dence of metabolic syndrome, hypertension, carotid artery thickening,and NAFLD in middle-aged and elderly Chinese adults with habitualdiets.

Funding SourcesThe National Natural Science Foundation of China (Nos.

81,472,965, 81,730,090, and 81,773,416).

Interaction of FADS2 and Dietary Essential Fatty Acid Intake onMetabolic Risk in Mice (P10-024)

Adam J Chicco, Peter Linde, Amanda J Evans, Lance Li Puma,and Connor MWhitaker

Colorado State University

Background: Polyunsaturated fatty acids (PUFAs) play importantroles in inflammation and cardiovascular and metabolic function.However, controversy exists regarding the effects of dietary PUFA intakeon metabolic risk owing to inconsistent results from studies in theliterature.

Objective: The aim of the present study was to address thehypothesis that variations in the metabolism of dietary essential PUFAsmodulate metabolic responses to a modern diet, as suggested by alteredcardiometabolic risk profiles in populations with polymorphisms in theFADS2 gene encoding delta-6 desaturase.

Methods: We developed mice with global heterozygous deletion(low), wild-type (WT; medium), or mild transgenic overexpression(high) of FADS2 expression.Micewere fed a standardized high-fat (40%kcal) diet containing dietary essential PUFAs at ∼7% kcal, providedmostly as linoleic acid (18:2n–6, LA; as corn oil, providing a 57:1 n–6:n–3 ratio), α-linolenic acid (18:3n–3; as flax oil, providing a 1:4 n–6:n–3ratio), or a 50:50 blend of each, providing 1:1 n–6:n–3 PUFAs. Effectson glucose tolerance were monitored over 4 mo, followed by analysis ofliver fat deposition and macrophage content.

Results: In mice withWT FADS2 expression, enrichment of a high-fat diet with n–6 PUFAs augmented glucose intolerance, weight gain,and hepatic inflammation, whereas n–3 PUFAs tended to attenuatethese effects. The same metabolic syndrome phenotype was augmentedby high (transgenic) FADS2 overexpression and attenuated by lowFADS2 (knockdown) expression compared withWT controls when thedietary n–3:n–6 PUFA composition was balanced 1:1, demonstratinga potent influence of FADS2 expression on metabolic phenotype andhepatic inflammation. However, a dietary imbalance of n–3:n–6 PUFAintake, either toward a higher n–3 or a higher n–6 PUFA intake, tendedto mitigate the respective effects of high and low FADS2 expression onglucose tolerance.

Conclusion: Taken together, these results support a complex inter-action of FADS2 expression and dietary PUFA intake onmetabolic risk,prompting a need for further study of FADS2 haplotypes as potentiallyrelevant criteria for individualized dietary recommendations.

Funding SourcesThis project is supported by AFRI Grant # 2014–0,6594 from the

USDA National Institute of Food and Agriculture.

Supplementation with Dietary Cholesterol Increases SerumEstradiol in Female Rats with Lower Energy Availability followinga 12-wk Treadmill Endurance Training Program (P10-025)

Lyra R Clark, Michael Dellogono, and Thomas Wilson

University of Massachusetts Lowell

Background: Exercise-associated menstrual dysfunction (EAMD)in female endurance athletes causes suppression of ovarian sexhormones. Detrimental health effects of EAMD include decreasedbone density, increased risk of musculoskeletal injury, altered neu-roendocrine function, and endothelial dysfunction. Cholesterol supple-mentation has been reported to increase sex hormones in sedentarypopulations.

Objective: The purpose of this study was to investigate the effectsof increased dietary cholesterol on ovarian hormone concentrations infemale rats during a treadmill (TM) endurance training program.

Methods: Female Sprague-Dawley rats (n = 29) consuming a stan-dard semipurified low-cholesterol (LC) diet were randomly assignedto a control group (C + LC, n = 15) or an exercise training group(EX + LC, n = 14) that performed TM running 40 min/d, 6 d/wk.At the study midpoint (end wk 6), rats were randomized to eitherreceive a high-cholesterol (HC) diet or remain on the LC diet. Fastingblood samples were collected at baseline, week 6, and week 12. Data arereported as means ± SDs or medians (IQRs).

Results: At end of week 6, the EX + LC group had lower bodyweight (338± 38.2 vs. 377± 59.9 g, P< 0.05), mean daily energy intake[53.5 (51.8, 58.1) vs. 67.9 (60.4, 69.1), P < 0.01], serum leptin [1.6 (1.0,2.6) vs. 2.2 (1.8, 5.4) ng/mL, P < 0.05], and serum progesterone [58.1(37.9, 62.7) ng/mL vs. 84.8 (55.9, 90.4) ng/mL, P < 0.05] comparedwith the C + LC group. There were no significant differences in serumestradiol between groups. At the end of week 12, the EX + HC grouphad lower body weight (337 ± 40.5 vs. 408 ± 83.2 g, P < 0.05) andserum leptin [0.8 (0.6, 2.3) vs. 2.4 (1.8, 6.1) ng/mL, P < 0.05] than theC+ LC group. Serum estradiol was higher in the EX+HC group [11.4(8.2, 13.3) pg/mL] than in the C + LC [5.1 (3.4, 8.1) pg/mL, P < 0.05]and EX + LC groups [4.2 (2.5, 7.2) pg/mL, P = 0.01]. There wereno significant differences between groups in energy intake or serumprogesterone.

Conclusions: Despite lower energy availability, female rats con-suming an HC diet during TM endurance training had higher serumestradiol than rats consuming an LC diet. Lower voluntary energyintake in exercise-trained rats despite higher energy expenditure mayindicate that lower energy availability in endurance-trained individualsmay be inadvertent.

Funding SourcesSupported by Egg Nutrition Center Graduate Fellowship Research

Grant (Award # 00,0000000018378).

Gender, Stressful Life Events, Sleep, and Obesity: A ProspectiveStudy of Young People in a British Columbia Cohort (P10-026)

Annalijn I Conklin,1 Sherry Guo,1 Christopher Yao,2 AlexanderCT Tam,1 and Christopher G Richardson1

1University of British Columbia, Canada; and 2Centre for HealthEvaluation and Outcomes Science, Canada


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Objectives: The aim of this study were to investigate 1) the gender-specific association between stressful life events and subsequent obesity,and 2) the potential modification of the effect by sleep deprivation.

Methods: We undertook a prospective study using population-based cohort data on self-reported obesity, stressful life events, sleepdeprivation, and covariables in adolescents aged 13–17 y (n= 916). Thedata were analyzed bymultivariable logistic regression, with interactionterms, and postestimation calculation of adjusted mean prevalence.

Results: Young men reporting multiple life events were more thantwice as likely to develop obesity at 6-mo follow-up (OR: 2.74; 95% CI:1.09, 6.89) as those reporting no events. Although more females than

FIGURE 1 Main association of stressful life events and predicted probability of being obese in young men (left panel) and young women(right panel).

FIGURE 2 Main association of stressful life events and predicted probability of being obese, by sleep deprivation, in young men (leftpanel) and young women (right panel).

males reported multiple stressful life events, we found no associationwith obesity in young women. We also found a more pronouncedassociation between stressful events and obesity among those reportingbeing sleep deprived (versus not deprived), particularly for young men.

Conclusion: Our findings highlight the importance of taking agender perspective when studying the development of obesity inadolescence, as well as the need for more robust longitudinal studieslooking at stressful life events and obesity in adolescence.

Funding SourcesThis research was partly funded by the UBC Work Learn Program

for undergraduate students.


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Gender, Ethnic Identification, and Obesity: A Prospective Studyof Young People in a British Columbia Cohort (P10-027)

Annalijn I Conklin, Alexander Tam, Shery XR Guo, andChristopher G Richardson

University of British Columbia, Canada

Objective: The aim of this study was to investigate the gender-specific longitudinal association between ethnic identification andobesity in young people.

Methods: We undertook a prospective study using population-based cohort data on self-reported ethnic identification (Multi-EthnicIdentification Measure), obesity, and covariables. Multivariable logisticregression using interaction terms for sex and predictors estimated ORsand postestimation calculated the adjusted mean prevalence.

Results: Young men reporting stronger overall ethnic identificationhad an∼50% greater likelihood of being obese at wave 6 (OR: 1.52; 95%

FIGURE 1 Main association of overall ethnic identification and predicted probability of being obese in young men (left panel) and youngwomen (right panel).

CI: 1.02, 2.25) than those with weaker identification. The monotonicassociation between ethnic identification and adjusted mean obesityprevalence was only seen in young men (Figure 1). Gender-specificfindings further differed by ethnic group (Figures 2 and 3), and thepatterns also depended on the component of the ethnic identificationexamined.

Conclusions: Although ethnic identification may offer protectivehealth-related quality of life effects, there may be more insalubriouseffects on other health outcomes, such as obesity, particularly for youngmen from certain ethnic groups. A greater understanding of the gender-specific role of ethnic identification in the development of obesity wouldbenefit public health efforts to use culture as an obesity interventionamong diverse young people.

Funding SourcesThis research was partly funded by the UBCWork Learn Program.


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FIGURE 2 Main association of overall ethnic identification and predicted probability of being obese, by Asian ethnicity, in young men(left panel) and young women (right panel).

FIGURE 3 Main association of overall ethnic identification and predicted probability of being obese, by Aboriginal ethnicity, in youngmen (left panel) and young women (right panel).

Adequacy of Protein Consumption 1 and 4 wk after HospitalDischarge in Older Adults (P10-028)

Rachel R Deer, Erin Maxwell, Kristen Poynor, Shawn Goodlett,and Elena Volpi

University of Texas Medical Branch

Background: Patients often experience nutritional decline duringhospitalization. This is attributable to a number of concomitant

conditions, including sleep disruption, pain, poor nutritional intake,physical deconditioning, and increased polypharmacy. Patients atrisk for undernutrition at discharge overlap with many risk factorsfor readmission. The Academy of Nutrition and Dietetics and theAmerican Society for Parenteral and Enteral Nutrition have called forthe development of care plans for this high-risk population. However,nutritional intake and meal patterns of older adults postdischarge havenot been well established.


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Objectives: This study aimed to determine the protein intake ofolder adults after an acute hospitalization and to compare these findingsto national recommendations.

Methods: Subjects (≥65 y old, n = 113) were recruited duringhospitalization at UTMB. Guided food recalls were collected at 1and 4 wk postdischarge using the Automated Self-Administered 24-hRecall in a subset of subjects (n = 34). Protein intake was comparedwith the current RDA (0.8 g · kg–1 · d–1), a higher proposed proteinrecommendation (1.2 g · kg–1 · d–1), and a per-meal recommendation(0.4 g · kg–1 · meal–1). One sample t tests were used to compare eachparticipant’s average intake to each recommendation.

Results: The mean ± SD protein consumption was 0.85 ±0.32 g/kg, which was not significantly different than the current RDAbut was significantly lower than the higher recommendation of 1.2 g ·kg–1 · d–1. It is important to note, however, that only 42% of participantsmet the RDA recommendation, and only 15% met the higher proteinrecommendation. The per-meal protein threshold recommendationwas met 0% at breakfast, 32% at lunch, and 47% at dinner.

Conclusions: We found that the majority of older adults are notconsuming the current RDA of protein after hospital discharge andeven less are meeting the higher recommendation for optimal proteinintake to compensate for the catabolic conditions experienced duringhospitalization. A skewed dietary protein distribution was present, withno participants meeting the recommended dietary protein threshold atbreakfast.More research needs to be conducted tomeasure the effects ofprotein supplementation, especially during breakfast, on postacute careoutcomes in older adults.

Funding SourcesFunding was from National Dairy Council (1229), NIH-National

Center for Advancing Translational Sciences (CTSA TL1R001440), andUTMB Claude D Pepper OAIC (5P30AG024832).

Impact of Ghanaian Vegetarian Diet on Male Endogenous SexHormones (P10-029)

Yauniuck Y Dogbe,1 Matilda Asante,2 and George Asare2

1No affiliation; and 2University of Ghana

Background:Avegetarian diet consisting of vegetables and legumes,and particularly soy products, is rich in phytoestrogens, which competewith other estrogens for receptor binding. For the male vegetarian,an increased serum concentration of estrogens may influence theconcentrations of testosterone and sex hormone-binding globulin. Thisis the first study to examine the concentrations of endogenous sexhormones in vegetarian and nonvegetarian males in Ghana.

Objective: The aim of the study was to examine the concentrationsof endogenous sex hormones in vegetarian and nonvegetarian males.

Methods: This was a case-control study of 52 apparently healthymales (22 vegetarians and 30 nonvegetarians) recruited from 4 locationsin the Greater Accra Region. A structured questionnaire was usedto obtain background and anthropometric data of all participants.Current and usual dietary intakes were assessed using a 24-h recalland a modified food frequency questionnaire, respectively. Bloodsamples were collected after an overnight fast and serum hormoneconcentrations were assessed by radioimmunoassay.

Results: The vegetarians were older than the nonvegetarians.The mean age of the participants was 49.4 ± 12.7 y. Motivation forchoosing the dietary lifestyle was more religious than health oriented.The vegetarians weighed less than the nonvegetarians, but their bodymass indexes were not significantly different. There were significantdifferences in mean intakes of protein (P= 0.001), calcium (P= 0.003),iron (P = 0.001), and zinc (P = 0.043). The mean serum testosteroneconcentration of the vegetarians (20.0 nmol/L) was significantly lower(P = 0.022) than that of the nonvegetarians (24.6 nmol/L), but themean serum estradiol concentration of the vegetarians (339 pmol/L)was significantly higher (P = 0.037) than that of the nonvegetarians(279 pmol/L). Significant differences (P= 0.001) were found comparingage and type of protein consumed with hormone concentrations.Besides estradiol in nonvegetarians, there were inverse but insignificantassociations of age and type of protein consumed with hormoneconcentrations.

Conclusion: There were significant differences in mean weight,diastolic blood pressure, and visceral fat between vegetarians and non-vegetarians.Mean serum testosteronewas significantly lower in vegetar-ians, but mean serum estradiol was significantly higher in vegetarians.

Funding SourcesSelf-funded.

The Effects of Breakfast Type and Habitual Breakfast Patterns onMorning Appetite and Satiety in Overweight Late-Adolescent Girls(P10-030)

Steve Douglas, Adam Byers, and Heather Leidy


Objective: The purpose of this study was to examine thepostprandial appetitive and hormonal responses to normal-protein(NP) and high-protein (HP) breakfasts in overweight adolescents whohabitually consume breakfast (H-C) and those who habitually skipbreakfast (H-B).

Methods: A total of 37 girls (age: 19 ± 1 y; body mass index:28.7 ± 0.5 kg/m2) participated in this semi-randomized crossover-design study. The participants were grouped according to the frequencyof habitual breakfast consumption. H-C (n = 19) and H-S (n = 18)consumed an NP (350 kcal; 12 g of protein) or HP (350 kcal; 32 g ofprotein) breakfast for 3 d. On day 4 of each pattern, a 5-h testing daywas completed. On this day, the respective breakfast was provided andpostprandial appetite and satiety sensations in combinationwith plasmagut peptides (PYY and ghrelin) were collected throughout themorning.

Results: A significant effect of time was observed in response tobreakfast for both H-C and H-S such that the consumption of breakfastdecreased hunger, desire to eat, prospective food consumption, andPYY and ghrelin concentrations, and increased fullness throughout the4-h postprandial period (all P < 0.01). Although no main effects oftreatment or habitual breakfast patternswere detected for any outcomes,treatment × habitual breakfast pattern interactions were observedfor plasma ghrelin concentration and prospective food consumption.Specifically, the 4-h postprandial ghrelin area under the curve washigher following the HP vs. the NP breakfast (24,900 ± 2570 vs.21,100 ± 2320 pg · ml–1 · min–1, respectively; P < 0.05) within theH-C group but not the H-C group. In addition, the 4-h postprandial


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prospective food consumption area under the curve was greaterfollowing the NP breakfast in H-C vs. H-S (6030 ± 672 vs. 8380 ± 682mm/min; P < 0.03).

Conclusion: Although differences in the ghrelin response to an NPvs. an HP breakfast meal and prospective food consumption followingan NP breakfast was influenced by habitual breakfast patterns, no otherdifferences in markers of appetite and satiety were observed in healthybut overweight late adolescent girls.

Funding SourcesNational Cattlemen’s Beef Association and the EggNutritionCenter.

Liver X Receptor, HDL, and HDL-Associated Proteins RegulateChronic Myelogenous Leukemia Cell Differentiation (P10-031)

Lydia Dupree, Kaley McMullen, Layra Cintron-Rivera, andCatherine J Andersen

Fairfield University

Objectives: We have previously demonstrated that liver X receptor(LXR) activation reduces the viability of human erythroleukemia K562cells—a model for chronic myelogenous leukemia, corresponding tochanges in cholesterol gene expression. Interestingly, human-derivedLDL, HDL, HDL-associated apolipoprotein A1 (apoA1), and proin-flammatory, serum amyloid A (SAA)-rich HDL exerted differentialeffects on K562 cell viability, both in the presence and absence ofLXR activation. In this study, we investigated whether human-derivedlipoproteins and LXR activation could further modulate markers ofK562 differentiation toward erythroid and megakaryocyte lineages.

Methods: K562 cells were cultured in the presence of humanHDL, LDL, apoA1, or SAA-rich HDL for 72 h, with or withoutcotreatment with the LXR agonist TO901317 (5 mM). K562 cells wereanalyzed for cellular cholesterol and hemoglobin content (normalizedto protein), in addition to mRNA expression of erythroid lineagemarkers, erythroid aminolevulinate synthase (ALAS-E), and y-globin,and the megakaryocyte lineage marker glycoprotein IIb (GPIIB).

Results: In the absence of lipoproteins, LXR activation reduced cel-lular cholesterol content, while increasing hemoglobin concentrations.We additionally observed increased mRNA expression of erythroid-specific ALAS-E and y-globin, and decreases in the megakaryocytemarker GPIIB. The addition of HDL and apoAI further increasedmRNA expression of y-globin under LXR activation, whereas ALAS-EmRNA expression was differentially increased by HDL, but decreasedby apoAI. Interestingly, SAA-rich HDL increased GPIIB mRNAexpression in the absence of LXR activation, whereas cotreatment withthe LXR agonist TO901317 ablated this effect.

Conclusion: These findings suggest that LXR activation, HDL, andHDL-associated proteins differentially regulate K562 chronic myeloge-nous leukemia differentiation toward erythroid and megakaryocytelineages.

Funding SourcesFairfield University College of Arts and Sciences.

Children’s Perceptions of the Importance of Limiting SugarSweetened Beverages and Strategies for Controlling Intake (P10-032)

Kaitlyn Eck,1 Kaitlyn Eck,1 Alek Dinesen,1 Colleen Delaney,1Karla Shelnutt,2 Melissa Olfert,3 and Carol Byrd-Bredbenner1

1Rutgers University, NJ; 2University of Florida; and 3West VirginiaUniversity

Objectives:The aim of this study was to describe children’s thoughtsrelated to sugar-sweetened beverages (SSBs) and their strategies forcontrolling intake.

Methods: School-aged children (aged 6–11 y) in 3 states (FL, NJ,andWV) completed surveys (n= 197) and participated in SSB-themedfocus groups conducted by trained moderators (n = 44). Two trainedresearchers continuously monitored data to identify the point of datasaturation and coded data to detect themes and trends.

Results: Surveyed children reported drinking SSBs 2.47 ± 2.44(mean ± SD) times a week. Focus group discussions revealed the SSBsthat the children most commonly drank were soda, sports drinks,lemonade, and fruit drinks. Children were more likely to drink sodaonly on weekends or at special events, whereas they drank sports andjuice drinks more often. Children thought it was important to limitSSBs to prevent cavities, diabetes, and feeling “sick.” Children also citedthe importance of drinking water for hydration, but few mentionedcalories or weight gain. Children preferred the taste of SSBs over otherbeverages, which made it hard for them to limit their intake. Childrenrelied on their parents to help them control intake by limiting thenumber of SSBs they were allowed, not purchasing SSBs, and buyingalternatives, like diet soda and water. Children felt parents could getchildren to drink fewer SSBs by talking to them about the negativehealth consequences of drinking toomany SSBs and rewarding them forhaving healthy drinks. Children also felt they could play a role in helpingtheir families drink fewer SSBs by telling family members that SSBsare unhealthy and encouraging them to drink milk or water instead.Children agreed that they mimicked their parents’ beverage choicesand wanted to drink whatever their parents had. Children recognizedthat their own beverage choices influenced what family members,particularly siblings, drank. Some children reported encouraging theirsiblings to select healthier beverages.

Conclusions: Elementary school-age children are aware of severalnegative health consequences associated with SSB intake, but not thepotential negative effects on weight. Children relied on parents to helpthem limit SSBs. Future nutrition interventions should help parentsunderstand how they affect children’s SSB intake through modelingbehaviors and setting limits.

Funding SourcesUSDA, National Institute of Food and Agriculture, Grant Number

2017-680,001-26,351.

Whole-Body Short-Chain Fatty Acid Production Is Not Dis-turbed in Patients with Chronic Obstructive Pulmonary Diseasewith Mild Gastrointestinal Symptoms (P10-033)

Marielle PKJ Engelen, Nicolaas Deutz, Sunday Y Simbo, andMarielle PKJ Engelen

Texas A&M University

Background: The short-chain fatty acids (SCFAs) acetate (C2),propionate (C3), and butyrate (C4), produced from fiber by thecolonic microbiota, are important for intestinal health as they are themain energy source of colonocytes and they positively influence guthealth–related processes. Gut problems are more abundant in patients


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with chronic obstructive pulmonary disease (COPD) than in healthyadults. However, SCFAmetabolism has not been investigated in COPDpatients.

Objective: The aim of this study was to examine whether distur-bances in whole-body SCFA production rates are present in patientswith COPD with mild gastrointestinal symptoms using a novel SCFApulse approach.

Methods: A pulse containing the stable isotopes of [1,2-13C2]acetate, [1-13C]propionate, and [1-13C]butyrate was administered intra-venously in 15 patients with moderate to severe COPD (GOLD stagesII–IV) and 14 healthy control subjects, and blood samples were takenfor 1 h. Plasma enrichments were measured as tracer-to-tracee ratios(TTR) by GC-MS/MS, and endogenous SCFA rate of appearances (Ra)were calculated from the area under the curve. Statistical analysis wasperformed by unpaired t tests or Mann-Whitney U tests. Relationshipsbetween Ra and disease-related characteristics, body composition, andfiber intake were analyzed with Pearson’s or Spearman’s correlationcoefficients. Data are expressed as means ± SEs.

Results:Therewere no differences in the Ra betweenCOPDpatientsand healthy subjects for acetate (426.7 ± 37.1 vs. 433.4 ± 41.1 μmol ·kg lean mass–1 · h–1, respectively), propionate (22.6 ± 2.1 vs. 21.8 ± 2.4μmol · kg lean mass–1 · h–1, respectively), or butyrate (10.2 ± 2.1 vs.9.3± 1.5μmol · kg leanmass–1 · h–1, respectively). The Ra of propionatewas higher in women than in men (25.9 ± 2.3 vs. 18.7 ± 1.5 μmol ·kg lean mass–1 · h–1, respectively) and positively associated with bodymass index (r = 0.69; P < 0.01). No significant relations were observedbetween any of the SCFA Ra and disease severity, body composition, ordietary fiber intake.

Conclusions: Whole-body production rates of the SCFAs acetate,propionate, and butyrate are not disturbed in COPD patients with mildgastrointestinal symptoms, but a gender effect exists for propionateproduction.

Funding SourcesNone.

Fermentability of Novel Type-4 Resistant Starches in In vitroSystem (P10-034)

Jennifer M Erickson,1 Justin Carlson,1 Maria L Stewart,2 andJoanne L Slavin1

1University of Minnesota; and 2Ingredion Inc.

Background: Resistant starches are nondigestible starches that arefermented in the colon bymicrobiota. These carbohydrates are prebioticand can be beneficial to consumer health. Many types of resistant starchexist with varying physical properties that may result in differences infermentability.

Objective: The objective of this research project was to comparefermentability of 4 novel resistant starches using an in vitro fermen-tation system by measuring changes in total gas production, pH, andformation of short-chain fatty acids (SCFAs).

Methods: Fecal donations were collected from 7 healthy volunteers.Four novel resistant starches—modified potato starch (MPS), modifiedtapioca starch (MTS), and two modified maize starches (MMS-1 andMMS-2)—were analyzed and compared with polydextrose and short-chain fructo-oligosaccharides as controls.

Results: After 24 h of fermentation, MPS and MTS respondedsimilarly in gas production (74 vs. 70.6 mL, respectively), pH (5.93vs. 5.93, respectively), and SCFA production (acetate: 115 vs. 124μmol/mL; propionate: 21 vs. 26μmol/mL; butyrate: 29 vs. 31μmol/mL,respectively). Although MMS-1 had similar gas production andindividual SCFAproduction, the pHwas significantly higher (6.06). Thefermentation ofMMS-2 produced the least amount of gas (22mL), witha higher pH (6.34) and lower acetate production (78.4 μmol/mL).

Conclusions: All analyzed compounds were fermentable andpromoted the formation of beneficial SCFAs.

Funding SourcesThis study was funded by Ingredion, Incorporated.

Protein-Restricted Diets Increase Energy Intake with VeryModerate Increase in Adiposity, Challenging the Protein LeverageHypothesis in Mice (P10-035)

Patrick C Even,1,2,3,4 Dalila Azzout-Marniche,1 Anne Blais,3Catherine Chaumontet,3 Gilles Fromentin,3 Claire Gaudichon,1and Daniel Tome1

1AgroParistech; 2UMRPNCA; 3INRA; and 4AgroParisTech,Univer-sité Paris-Saclay

Background: Protein-deficient diets are suspected to increase foodand energy intake to compensate for protein deficiency, which increasesthe risk of fat deposition and adiposity. This process can be exacerbatedspecifically by proteins with a limited methionine content, such as soy.

Objectives: The aim of this study was to investigate food intake,adiposity, energy expenditure, and related biomarkers in mice fed a soyor casein protein diet.

Methods: After 2 wk of adaptation, female BALB/c mice (8 wk old)were assigned to 6 different diets for 8 wk (n= 8 per group), with either20%, 6%, or 3% casein (P20C, P6C, P3C), 20% or 6% soy protein (P20S,P6S), or 6% soy protein with corrected methionine concentrations tothe P6C diet (P6S-Cor).

Results: Energy intake was significantly greater in P3C comparedwith P20C mice and in P6S compared with P20S mice. Final bodyweight, lean body mass, and bone mineral density were lower onlyin P3C compared with P20C mice. Total fat was lower in P20S(1.1 ± 0.09 g) than in P20C (1.7 ± 0.15 g) and P6S (1.9 ± 0.21 g) mice(P < 0.05), whereas fat mass was similar in P20C, P6C, P3C, P6S, andP6S-Cor mice. Total energy expenditure was greater in P3C comparedwith P20Cmice and in P6S compared with P20Smice. Gene expressionwas unchanged in white adipose tissue and brown adipose tissue, andwas marginally changed in liver (FAS and CD36) and muscle (Ulk1). Inthe hypothalamus,mRNA expressionwas greater forNPY and lower forPOMC in P3C than in P20C, P6C, P20S, P6S, and P6S-Cor fed mice.

Conclusions: Low-protein soy or casein diets increased food intakebut also increased energy expenditure, which strongly moderated thegain in fat mass expected from the larger energy intake. Themethionine-deficient soy protein diet has a specific and independenteffect on adiposity. These results challenge the protein leveragehypothesis that a low-protein diet can induce significant increases infood intake and in body adiposity, and confirm the role played by thedietary methionine content on the regulation of body adiposity.

Funding SourcesINRA, AgroParisTech.


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FIGURE 1 Energy intake adjusted to 20 g of AMM. Bars with a different letter are significantly different (P < 0.05).

FIGURE 2 Total and resting energy expenditure in the different mouse groups. Bars with a different letter are significantly different(P < 0.05).


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FIGURE 3 mRNA expression of muscle Ulk1, liver FAS, and CD36, and hypothalamic mRNA expression NPY/POMC ratio in the differentmouse groups. Bars with a different letter are significantly different (P < 0.05).

TheMetabolic Availability of Lysine in Cooked Millet in HealthyAdult Men as Shown by the Indicator Amino Acid OxidationTechnique (P10-036)

Abrar Fakiha,1 Paul Pencharz,2 Ronald Ball,3 Rajavel Elango,4and Glenda Courtney-Martin2

1University of Toronto, Canada; 2The Hospital for Sick Children,Canada; 3University of Alberta; and 4BC Children’s Hospital

Objective: The objective of this study was to measure the proteinquality of cooked pearl millet by determining the metabolic availability(MA) of its most limiting indispensable amino acid, lysine. In addition,we aimed to study the effect of complementing millet with lentils on theMA of lysine in a mixed meal format.

Methods: The MA of lysine in millet was studied using theindicator amino acid oxidation method (IAAO) technique with L-[1-13C]phenylalanine as the indicator. In order to determine the MA oflysine, 4 concentrations of lysine were tested [5, 8, 12, and 15 mg ·kg–1 · d–1, which represent 13%, 21%, 32%, and 40% of the lysinerequirement for adults (37 mg · kg–1 · d–1)] in 5 healthy adult men in arepeated-measures design. Each subject participated in 8 experiments:4 lysine intakes as L-lysine from crystalline amino acid, 3 intakes oflysine from millet prepared by moist cooking, and 1 experiment fordietary complementation with lentils. The MA of lysine was estimatedby comparing the IAAO response to varying intakes of lysine in cookedmillet compared with the IAAO response to lysine intakes in the

reference protein (crystalline amino acid mixture patterned after eggprotein) using the slope ratio method.

Results: Preliminary data from 3 of the 5 subjects show an MA oflysine frommillet of 50%. Analysis of the complementation with lentilsis incomplete at this time.

Conclusion: Our current data suggest that diets based upon milletas the protein source are severely limiting in bioavailable lysine andtherefore the protein quality of diets containing millet are much lowerthan previously believed.

Funding SourcesFunded by the Canadian Institute of Health Research (FRN 148,864)

and a Saudi Arabian Government scholarship.

The Role of RGS10 in the Serum Lipid Profile and HepaticAntioxidant Defenses in Mice Fed a High-Fat Diet (P10-037)

Xi Fang, Hea Jin Park, Jae-Kyung Lee, Jaegwon Chung, ErikOlsen, and Julie Jeon

University of Georgia

Objective: Regulator of G protein signaling 10 (RGS10) has shownto negatively regulate inflammatory response and ameliorate impairedglucose metabolism induced by a high-fat diet (HFD). To betterdetermine the role of RGS10 in the progress of chronic diseases, wehypothesized that RGS10 depletion will result in abnormal lipid profile


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and compromised antioxidant defenses which will be exaggerated by anHFD. The aim of this study was to test that hypothesis.

Methods: RGS10 knockout (KO)mice were fed a low-fat diet (LFD)or an HFD for 8 wk (n = 5–8/group). C57BL/6J wild-type (WT) micewere fed LFD as a control. Serum lipid profile, hepatic glutathioneredox status, and activity of hepatic antioxidant enzymesweremeasuredfollowing the treatments.

Results: We have previously observed that KO mice showedsignificantly greater body weight gain and adiposity following HFDcompared with WT mice. In the current study, KO mice had higherserum triglyceride (TG) concentration than WT mice, whereas serumcholesterol concentration was similar in the 2 groups. In KO mice,HFD increased serum cholesterol concentration by 2.3 times but didnot influence serum TG concentration compared with LFD. Hepaticreduced glutathione (rGSH) of KOmice was similar to that ofWTmice,whereas hepatic oxidized GSH tended to be higher in KO mice. HFDsignificantly decreased the total GSH in KOmice, suggesting that HFDmay decrease GSH synthesis in the liver of KO mice. KO mice tendedto have lower activity of catalase than WT mice, which potentiallyincreases oxidative stress. The activity of superoxide dismutase (SOD),another major antioxidant enzyme detoxifying reactive oxygen species,did not differ between groups. Hepatic rGSH showed a negativecorrelation with serum cholesterol and SOD and a positive correlationwith catalase. These results demonstrate that the disrupted lipidmetabolism may be related to the elevated oxidative stress in KO mice,which can be exaggerated by HFD.

Conclusions: KO mice displayed impaired antioxidant defensesand imbalanced glutathione redox status, which may lead to elevatedinflammation. HFD exaggerated the impaired antioxidant defenses inKO mice, which may be related to increased adiposity and excessiveweight gain.

Funding SourcesHJP was supported by Georgia Experimental Agricultural Station,

HATCH #GEO00795, and College of Family and Consumer Sciences.JKLwas supported by startup funds from theDepartment of Physiologyand Pharmacology, College of Veterinary Medicine at UGA.

Identification of Enzymes Responsible for Endogenous Synthesisof Erythritol from the Pentose Phosphate Pathway (P10-038)

Martha S Field,1 Lisa Krämer,2 and Karsten Hiller2

1Cornell University, NY; and 2Technische Universität Braunschweig

Background:Nontargetedmetabolomics experiments performed ina cohort of young adults followed over the course of their first year ofcollege identified erythritol as a novel biomarker of central adipositygain. Erythritol concentrations in pooled plasma from individualsexhibiting central adiposity gain were nearly 15-fold higher than inplasma from individuals with stable adiposity and nearly 23-fold higherin individuals with higher glycemia compared with those with lowerglycemia. Erythritol is a 4-carbon sugar and a low-calorie sweeteneror food additive that is rapidly absorbed then excreted in urinefollowing ingestion. Stable isotope tracer experiments demonstratedthat erythritol is formed by the reduction of erythrose, which isproduced from erythrose-4-phosphate, a pentose phosphate pathwayintermediate.

Objective: The objective of this study was to determine whichenzymes are responsible for conversion of erythrose to erythritol.

Methods: This study used column chromatography to purify 2distinct enzyme activities from rabbit liver, which proteomic analysisidentified as alcohol dehydrogenase 1 (ADH1) and sorbitol dehydroge-nase (SORD). Recombinant human ADH1 (4 variants) and SORDwereexpressed, purified, and kinetically characterized.

Results: Both human recombinant ADH1 and SORD reduceerythrose to erythritol, using either the reduced form of nicotinamideadenine dinucleotide phosphate (NADPH) or the reduced form ofnicotinamide adenine dinucleotide as a cofactor. Interestingly, ADH1variants vary with respect to both their affinity for erythrose andturnover number. Cell culture studies indicated that this activityis primarily NADPH-dependent in vivo. Interestingly, the ADH1B2variant, common in Asian populations, is not active when NADPH isused as a cofactor.

Conclusions: ADH1 catalyzes the first step of ethanol metabolism,and is known to have several human population-specific variantsthat markedly affect enzyme activity and alcohol metabolism capacity,suggesting that these variants may also account for population-specificvariations in erythritol production from glucose and influence glucoseutilization and central metabolism.

Long-TermUse ofRELiZORBbyChildren andAdults withCysticFibrosis Receiving Enteral Feeding Increases Omega-3 Fatty AcidLevels and Improves Nutritional Parameters (P10-039)

Steven Freedman,1 John Stevens,2 Colby Wyatt,3 Perry Brown,4Danica Grujic,5 and Eric First5

1Beth Israel Deaconess Medical Center, MA; 2Riley Hospital forChildren, Indiana University School of Medicine; 3Maine MedicalCenter; 4St. Luke’s Health System,MO; and 5Alcresta Therapeutics, Inc.

Background:Pancreatic insufficiency (PI) andmalabsorption of fatslead to reduced caloric intake, inability to maintain or gain weight, andincreased gastrointestinal symptoms. As a result, enteral feeding (EF)is used in patients with cystic fibrosis (CF) who have poor nutritionalstatus. In addition to total fat malabsorption, low concentrations oflong-chain polyunsaturated fatty acids, such as the omega-3 (n–3) fattyacids docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA),have been shown to be decreased in patients with CF and may play arole in the pathogenesis of inflammation in CF. RELiZORB (AlcrestaTherapeutics) was developed to hydrolyze fats in EF prior to ingestion.In an initial trial in patients with CF on EF, REliZORB was shown todecrease gastrointestinal symptoms and significantly increase DHA andEPA concentrations in plasma.

Objective: The purpose of the current study was to examine thesafety, tolerability, and fat absorption, using a long-term measure of fatuptake reflected as DHA + EPA concentrations in RBC membranes, inchildren and adults with CF, with the use of RELiZORB over 90 d.

Methods: Subjects aged 5–33 y (n = 39) enrolled in a multicenteropen label study. The study included a 7-d run-in period, followedby a 90-d treatment period, during which RELiZORB was used withovernight EF (Impact Peptide 1.5). PERTs were used with solid foods,but not with overnight EF. Safety assessments were taken during thestudy. Outcomes examined included RBC uptake of DHA + EPA (theprimary endpoint); change from baseline in plasma concentrations of


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FAs, vitamins, and weight were also assessed. Blood was obtained atstudy entry, baseline, and treatment days 30, 60, and 90.

Results: A total of 36 subjects completed the study (22 males). Themean age was 13.6 y, the mean body mass index was 19.9 and the meanuse of EF was 6.5 y. The mean formula volume was 749 mL, with afrequency of 5.6 uses a week. RELiZORB use was well tolerated, withno unanticipated adverse events. Fat absorption significantly improved,as shown by increased RBC concentrations of DHA + EPA, improvedω-6/ω-3 fatty acid (FA) ratio, and increased plasma concentrations ofDHA + EPA as early as 30 d of treatment.

Conclusions:RELiZORBusewas found to be safe andwell tolerated,and resulted in increased concentrations of long-chain polyunsaturatedfatty acids in RBCs and plasma. This is the first prospective study toshow that EF can improve FA abnormalities in CF. Since improvementinω-6 FAs has been shown to help pulmonary and inflammatory status,and anthropometric parameters in CF, RELiZORBmay have importantlong-term therapeutic benefits in patients with CF.

This trial was registered at clinicaltrials.gov as NCT02750501.Funding SourcesThis study was sponsored by Alcresta Therapeutics, Inc.


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Lower Dairy Intake Affects Blood Pressure but Not MetabolicSyndrome in Community-Based Adults (P10-040)

Patricia Baston Frenhani,1 Tatiana Figueira,2 Silvia JustinaPapini,2 and Roberto Carlos Burini2

1School of Nutrition—Centre for Life Sciences Courses (CCV)—PUC-Campinas, Brazil; and 2Unesp Medical School, Brazil

Objective: The aim of this study was to investigate the associationof dairy intake with metabolic syndrome (MetS) in community-basedadults.

Methods:Cross-sectional analysis of baseline data from763 individ-uals, both genders, 54 ± 11 y old, was undertaken. The data collection(2006–2016) included individuals clinically and ethically selected forthe lifestyle modification program “Move for health,” and includeddata of socioeconomic, demographic, physical activity, and dietaryassessments, as well as the anthropometric, clinical, and biochemicaldata required for MetS diagnosis (NCEP-ATP III criteria). The dairyintake calculated in a 24-h food intake recall was processed by NutWin,version 1.5, and the results were evaluated by the Healthy Eating Index(HEI) score adapted for the Brazilian population. Statistical analysiswas performed using SAS 9.3 software, adopting the significance levelP < 0.05.

Results: The sample comprised 81.1% females, 75.2% were earningup to 5 family minimum wage, 65.4% self-referred good health, but49%were obese (two-thirds with abdominal obesity and one-third withMetS). Daily energy intake was 1525 ± 609 kcal (51.9% protein and30.2% fat). Dairy intake reached 92.5% of adequacy , and calcium intakereached 91.3% (589 ± 382 mg/d). Overall, only 3.5% of the sampleshowed an HEI >100 points (a good-quality diet). The lowest decileof dairy (p10) was associated with lower income, lower intake of totalenergy, fat, and calcium, and lower HEI. However, after adjustments,only calcium intake remained associatedwith the covariables. The lowerdairy (and calcium) intake was associated with hypertension (present in33.3% of the sample), but not with the presence of MetS. The lowestdecile of dairy intake had a 51% risk for hypertension, whereas p90of dairy presented a protective effect of 26%. The isolated protectiveeffect of calcium (OR: 0.96–0.98) against hypertension was confirmedby an intake with a higher calcium:protein ratio, without any influenceof sodium or potassium and/or carbohydrate or fiber of the diet.

Conclusions: The lower intake of dairy we found reflected (throughlower calcium input) hypertension rather than the MetS rate.

Funding SourcesSupported by CNPq and CAPES.

Is Water Intake with a Meal Associated with Overweight? EarlyResults from a Controlled Dietary Study (P10-041)

Elisa Fuscà, Gianluca Piras, Daniele Bottigliengo, EglePerissinotto, Ileana Baldi, and Dario Gregori

Department of Cardiac, Thoracic and Vascular Sciences, Universityof Padova, Italy

Background: The daily water intake, recommended by the Foodand Nutrition Board, is 2.7 L for women and 3.7 L for men, includingfood and beverages other than pure water. The association betweenwater intake and overweight or obese status in healthy people has

rarely been investigated, owing to difficulties in managing dietarycomposition at meals.

Objectives: We aimed to analyze the association between themain anthropometric measurements and water intake at meals, takingadvantage of the controlled dietary setting of the NOTION (Building ofa system to measure the caloric intake into the population) experiment.

Methods: A total of 30 young adult volunteers were recruited usingas selection criteria the absence of pathologies and special dietary habits.A questionnaire on physical activity was submitted to them. Height andweight weremeasured to get the bodymass index (BMI); BD—and con-sequentially fat-free mass, fat mass, and percentage body fat—were cal-culated by Durning andWomersley equations using 4 skinfolds (biceps,triceps, subscapularis, and iliac) and the Siri equation. The waist-to-hip ratio (WHR) was calculated by measurement of the hips and waistcircumferences. Water intake was measured during 4 meals, in a fullycontrolled dietary feeding intervention designed to simulate a normaland healthy diet, obtaining the total amount of water intake during 1 d.

Results: Water intake at meals was about the same for overweightor obese people as for normal weight people (579.67 ± 191.37 vs.510.88 ± 254.60 ml/d, respectively; P = 0.5430). The absence of anyassociation was also noticed by regression analysis, both unadjusted(Wald test, P = 0.3743) and adjusted with sex and BMI considered asconfounding factors (males to females: P = 0.1690). Figure 1 shows theassociation of weight and WHR with water intake.

Conclusions: This study suggests that there is a lack of evidenceof a relation between greater water intake and higher or lower BMI.Therefore, further investigations and work are required before anyimplications can be drawn regarding the implementation of dietaryinterventions.

Funding SourcesThe National Academy of Sciences Engineering Medicine

FIGURE 1 Association of weight and WHR with water intake.


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A Regulatory Insertion-Deletion Polymorphism in the FADSGene Cluster Determines PUFA Synthesis and Influences LipidProfiles (P10-042)

Ying Gao and Peiqin Li

Chinese Academy of Science

Background:Fatty acid desaturase 1 (FADS1) catalyzes the synthesisof ω-6 (n–6) arachidonic acid (AA) and ω-3 eicosapentaenoic acid(EPA). Polymorphism of rs66698963, a 22-bp insertion-deletion (Indel)137-bp downstream of a sterol response element in FADS2 intron 1,mediates expression of FADS1 in vitro, as well as exerting positiveselection in several human populations. The associations betweenrs66698963 polymorphism and plasma polyunsaturated fatty acids(PUFAs), as well as disease phenotype, are unclear.

Objective: This study aimed to evaluate the relation betweenrs66698963 genotypes and plasma PUFA concentrations, as well asblood lipid profiles.

Methods: The baseline plasma fatty acids were measured by GC ina population of 1504 healthy Chinese adults who were aged 35–59 y.Blood lipids were measured at baseline and after 2 y follow-up. Thers66698963 genotype was determined using agarose gel electrophoresis.Linear regression and logistic regression analyses were performed toassess the association between genotype and plasma PUFAs and bloodlipids.

Results: A shift from precursor to product PUFAs was observedfrom D/D to I/D to I/I genotypes for both ω-6 and ω-3 PUFAs. ForI/I compared with D/D carriers, plasma concentrations of ω-6 AA andthe ratio of AA to ω-3 EPA plus docosahexaenoic acid (DHA) were57% and 32% higher, respectively. Carriers of the deletion (D) allele ofrs66698963 had higher triglycerides (β = 0.029, P = 0.02) and lowerHDL cholesterol (β = –0.012, P = 0.02) than carriers of the insertion(I) allele.

Conclusions: The rs66698963 genotype controls AA and AA toEPA + DHA concentrations, reflecting the basal risk for inflammatoryand related chronic disease phenotypes, and is correlated with risk ofdyslipidemia.

Funding SourcesThe National Key Research and Development Plan

(2016YFD0400200) and The 100 Talented Plan of Chinese Academy ofSciences.

Bioelectrical Capacitance IsAssociatedwith InsulinResistance inNondiabetic Women, but Not Men (P10-043)

Valene Garr Barry and Barbara Gower

University of Alabama at Birmingham

Background: Recent evidence suggests that bioimpedance devicescan noninvasively detect physiologic dysfunction at the cellular levelby measuring changes in membrane capacitance (CM), an impedanceparameter that reflects membrane structure and integrity. However, it isnot known whether CM is influenced by insulin resistance, a commonantecedent of chronic metabolic diseases such as type 2 diabetes andcardiovascular disease.

Objectives: The aim of this study was therefore to test thehypotheses that 1) CM would be significantly higher in nondiabetic

insulin-resistant adults compared with insulin-sensitive adults, and 2)associations would exist between CM and endpoints reflecting glycemicstatus.

Methods: This cross-sectional study used a subsample of nondia-betic adults, 18–49 y old (n = 2212, 45% female, 43% non-Hispanicwhite, 22%non-Hispanic black, and 34%Hispanic) from the 1999–2004NHAMES III. Subjects were divided into groups by sex and homeostaticmodel assessment insulin resistance index (HOMA-IR; > or ≤2.6).ANCOVA with the Tukey-Kramer post hoc test was used to comparedifferences between groups. Bivariate and partial correlations wereused to determine associations between CM and endpoints reflectingglycemic status [glycated hemoglobin (HbA1c), fasting insulin,HOMA-IR]. Covariates included age, race, height, fat and lean mass from dual-energy X-ray absorptiometry, and medication use.

Results: In women, CM was associated with HbA1c (%) (r = 0.34;P < 0.0001), fasting insulin (µU/ml) (r = 0.46; P < 0.0001), andHOMA-IR (r = 0.47; P < 0.0001). After adjusting for covariates,insulin-resistant women had higher a CM than insulin-sensitive women(8.19 ± 0.07 vs. 8.80 ± 0.12 nF/m2; P < 0.01). Conversely, in men, CM

was not associated with HbA1c (r = 0.05; P = 0.08), but was associatedwith fasting insulin (r = 0.10; P < 0.01) and HOMA-IR (r = 0.19;P < 0.01). After adjusting for body composition, insulin-resistant menshowed no significant difference in CM compared with insulin-sensitivemen (11.28 ± 0.10 vs. 11.27 ± 0.11 nF/m2; P = 0.95).

Conclusions: CM may reflect insulin resistance in nondiabeticwomen, but not men. Future research will 1) explore the observedgender differences in CM, and 2) evaluate the association between CM

and insulin sensitivity measured by the euglycemic clamp.Funding SourcesT32HL105349—UAB Pre-Doctoral Training in Obesity-Related

Research from NHLBI (National Heart Lung and Blood Institute).

Digestive Bioavailability of Spirulina Protein andAminoAcids ina Rat Model (P10-044)

Claire Gaudichon, Nadezda Khodorova, Juliane Calvez, JulienPiedcoq, Daniel Tomé, and Romain Tessier

UMR PNCA, INRA, Agroapristech

Objectives: Spirulina is often used as a dietary supplement. Proteinbioavailability seems to be relatively low, but there have been few invivo studies. Our study aimed to measure the digestibility of spirulinaprotein and amino acids (AA) in vivo, and to determine whether it canbe improved by sonication.

Methods: A cohort of 12 males Wistar rats received a single mealcontaining 0.77 g of spirulina (0.44 g of protein) intrinsically labeledwith 15N. Six rats were given spirulina that had been sonicated for1.5 h and 6 were given untreated spirulina. Rats were killed 6 hafter meal ingestion and their gastrointestinal contents (stomach, smallintestine, ileum, cecum, and colon) were collected. Digestibility ofspirulina protein was determined by measurement of nitrogen and 15Nenrichment in the gastrointestinal contents by use of an elementaryanalyzer coupled to an isotopic ratio mass spectrometer. In order todetermine amino acid digestibility, amino acids were measured byHPLC and their 15N enrichments were measured by GC-combustion-isotopic ratio mass spectrometry. The digestible indispensable aminoacid score (DIAAS) was calculated as follow: DIAAS % = 100 × [(mg


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of digestible dietary IAA in 1 g of the dietary protein)/(mg of the samedietary IAA in 1 g of the reference protein)]. The results are expressedas mean ± SEM and compared by a t test (P < 0.05).

Results: At 6 h after ingestion, most of dietary nitrogen was foundin the cecum and the colon. Orofecal digestibility of untreated spirulinawas low (81.8% ± 1.8%). Sonication tended (P = 0.08) to improve it(86.9% ± 1.8%). Amino acid digestibility varied from 76.5% ± 2.1%for glycine to 90.5% ± 1.2% for methionine. There was no effect ofsonication onAAdigestibilities. DIAAS calculation shows that spirulinais only marginally limiting in lysine (0.98 ± 0.02). For the otherindispensable amino acids, the DIAAS score was >1.

Conclusions: This study shows that spirulina has a low digestibilitycompared with other proteins that were evaluated in the sameconditions. Based on amino acids measured, spirulina satisfies therequirements for amino acids. Prior sonication only tended to improvespirulina digestibility. Data on additional animals are currently beingcollected to confirm or refute this trend.

Funding SourcesAgence Nationale de la Recherche (ANR Prodige).

FIGURE 1 Dietary nitrogen recovered in gastrointestinalsegments 6 h after the meal.

Gene Expression in Differentiated 3T3-L1 Cells: Effect of GingerExtract (P10-045)

Khadija Ghanam,1 Xin Yi,2 Jayant Deshpande,2 and VijayaJuturu2

1Omniactive Health Technologies (Canada) Limited; and2OmniActive Health Technologies Inc.

Background: Ginger has gained importance recently for thetreatment of obesity and its complications. The gingerols and shogaolsin ginger extract (GE) may have thermogenic activity.

Objective: The objective of this study was to assess in vitro and invivo the effects of SCE GE on adipogenesis and thermogenesis. Obesityis the result of increased adipogenesis and downregulation of lipolysis.The differentiation of preadipocytes intomature adipocytes is regulatedby different transcription factors, such as peroxisome proliferator–activated receptors (PPARs) and CCAAT/enhancer binding proteins(C/EBPs).

Methods: We investigated the effects of GE on lipogenesis andthermogenic gene expression using an in vitro 3T3-L1 mouse cellmodel. After 6 d of treatment with GE (50 µg/ml) during adipocytedifferentiation, 3T3-L1 cell lysates were harvested and real-timepolymerase chain reaction was performed to evaluate the expressionof the adipogenic and thermogenic marker genes. Among the genesselected, C/EBP α and PPAR γ are the main regulators of adipogenesisand the key transcription factors that are responsible for the activationof adipogenic genes. Acetyl-CoA carboxylase α(ACCA), fatty acidsynthase (FASN) and stearoyl-CoA desaturase 1 (SCD1) are involvedin the lipogenic process, DLK1 is a negative regulator that inhibitsadipogenesis, and PGC1 α, situin 1, and uncoupling protein 1 areinvolved in thermogenesis. All statistics were analyzed using GraphPadPrism. Data were expressed as mean values ± SEMs. Results wereconsidered significant at P < 0.05.

Results: Our results showed that GE treatment significantly down-regulated ACCA, FASN, SCD1, PPAR γ , and C/EBP α gene expressionand slightly upregulated PGC1 α expression compared with the control.The FASN gene regulates fatty acid synthesis and lipid accumulation inliver and adipose tissue. SCD1genes catalyze the conversion of saturatedfatty acids into monounsaturated fatty acids. PGC1 α genes are relatedto the process of thermogenesis.

Conclusions: These results suggest that GE treatment promoteshigh expression of these genes and helps consume excess energy asheat and decrease the storage of energy excess as fat. The underlyingmolecular mechanism for this reduction is related to the inhibition ofadipogenesis and an upregulation of thermogenesis.

Funding SourcesOmniActive Health Technologies Ltd., India.

Protein-to-Energy Ratios over the Course of the Obesity Epi-demic; Analysis of AustralianNational Surveys from1983, 1995, and2012 (P10-046)

Amanda Lee Grech, Anna Rangan, and Margaret Allman-Farinelli

The University of Sydney

Background: The protein-leverage hypothesis proposes that ap-petite is tightly regulated to ensure that protein requirements aremet even if energy requirements have been satisfied. Therefore, dietswith a lower protein-to-energy ratio may contribute to excess energyintake, weight gain, and the increased prevalence of obesity. Obesity inAustralia has increased from 9% in 1983 to 18.7% in 1995, 27.5% in2012, and 27.9% in 2015.

Objectives: The primary aim of this analysis was to assess theprotein-to-energy ratio along with other macronutrients of a represen-tative sample of the Australian populations’ diets in 1983, 1995, and2012. The secondary aim was to determine if higher protein density iscorrelated with energy intakes.

Methods: Cross-sectional, national nutrition surveys from 1983,1995, and 2011/12 (total n = 16,118) assessed diet with 24-h recalls.Protein intake, adjusted for energy intake, and protein-to-energy ratios,expressed as the percentage energy from protein, were assessed in eachsurvey. Percentage energy from carbohydrate and fat was also assessed.Regression was used to assess energy intake with protein, adjusted


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for age, sex, and survey. Low energy reporters (energy intake to basalmetabolic rate ratio <0.87) were excluded.

Results: Protein intake was relatively stable, decreased from 98.9 gin 1983 by 2.7 g in 1995 and increasing by 3.5 g in 2011/12 (P < 0.001).Percentage energy from protein was 17.2%, 16.6%, and 17.9% in 1983,1995, and 2011/12, respectively (P< 0.001). Energy from carbohydratesincreased from 40.7% in 1983 to 45.4% and 43.7% (P< 0.001), whereasenergy from fat decreased between each survey, from 35.5% to 32.0% in1995 and to 31.2% in 2011/12 (P < 0.001). Protein density predictedenergy intake, and for each gram of protein per 1000 kJ, the energyintake decreased by 162 kJ (β = –162.03; P < 0.001).

Conclusions: Protein to energy ratios decreased between 1983 and1995 owing to higher carbohydrate intake and may have contributed tothe doubling of obesity in Australia during this time. The highest ratiosoccurred in 2011/12, at which point the obesity rate stabilized. Higherprotein density was correlated with lower energy intake, and may be animportant factor in the prevention of weight gain.


Obesity Promotes Global DNA Hypermethylation and DNAOxidative Damage in Liver of Female Zucker Rats (P10-047)

Reza Hakkak,1 Soheila Korourian,2 Oleksandra Pavliv,3 andStepan Melnyk3

1Arkansas Children’s Research Institute, University of Arkansas forMedical Sciences; 2University of Arkansas for Medical Sciences; and3Arkansas Children’s Research Institute

Background: The obesity epidemic has continuing to grow inthe United States and throughout the world in the last 2 decades.There is a link between obesity and the development of chronicdiseases such as diabetes, cardiovascular disease, certain types ofcancer, and liver diseases. Previously, we reported that obesity causeda significant increase in oxidative and nitrosative stress, oxidative DNAdamage, and changes of DNA methylation in the liver of the 7,12-dimethylbenz[a]anthracene-induced mammary tumor model.

Objective: The objective of the present study was to investigate theeffects of obesity on global DNAmethylation and DNA oxidative DNAdamage in the rat liver of a noncarcinogen-induced mammary tumormodel.

Methods: After 1 wk of acclimation, 5-wk-old female lean andobese Zucker rats (n = 8/group) were fed AIN-93-G diet for 8 wk.The rats were weighed twice a week. The concentration of liver samplemetabolites and DNAmodified bases were measured using HPLC withelectrochemical detection and LC-MS.

Results: Our results shows that obese rats: 1) gained significantly(P < 0.001) more body weight; 2) in liver, had a significantly(P < 0.05) lower concentration of reduced free glutathione (GSH), asignificantly (P < 0.04) higher concentration of oxidized glutathione(GSSG), a significantly lower (P < 0.03) GSH:GSSG oxidative ratio, asignificantly (P < 0.03) increased concentration of SAM, a significant(P < 0.001) decrease in SAH concentration, and a significantly(P < 0.001) increased SAM:SAH methylation ratio; and 3) in liverDNA, had a significantly (P < 0.03) increased concentration of 8-hydroxy-guanosine, a significantly (P < 0.03) increased concentration

of 5-hydroxymethyl-cytosine, and a significantly (P < 0.02) increaseconcentration of 5-methyl-cytosine.

Conclusion: Our results show that obesity promoted higher oxida-tive stress in liver, greater oxidative DNA damage, and global DNAhypermethylation in liver compared to lean rats. These changes maycontribute to gene expression modification, which, in turn, will lead toincreased risk of chronic liver disease development.

Funding SourcesThe project was funded by the Arkansas Children’s Research

Institute’s University Medical Group Fund grant program and theArkansas Bioscience Institute.

Omega-3 Fatty Acids Decrease Appetite and Increase Postpran-dial Energy Expenditure and Fat Oxidation in Normal Weight andOverweight or Obese Children (P10-048)

Aubree L Hawley,1 Elisabet Borsheim,2 Charlayne F Mitchell,1Sarah L Russell,1 HexiruiWu,1 Eva Dehaene,3 and Jamie I Baum4

1University of Arkansas; 2University of Arkansas for MedicalSciences, Arkansas Children’s Nutrition Center; 3University of Gent,Belgium; and 4University of Arkansas, Center for Human Nutrition

Objectives:The objective of this studywas to determine if increasingO3FA at breakfast increases the thermic effect of feeding (TEF) anddecreases food intake in overweight and obese children (OW).

Methods: In total, 20 male and female, normal weight [NW; n= 11,age: 9.8 ± 0.4 y; body mass index (BMI) percentile corrected for ageand sex: 49.5% ± 6.9%] and OW (n = 9; age: 10.2 ± 0.5 y; BMIpercentile: 91.1 ± 1.5%) children were recruited to participate in thisrandomized, crossover, double-blind study. Participants consumed atest beverage high in O3FA [5 g of O3FA (2 g of eicosapentaenoicacid; 2 g of docosahexaenoic acid)] and an isocaloric (320 kcal),macronutrient-matched control beverage (with 5 g of corn oil) witha washout period of ≥4 wk between test days. Participants arrivedfasted, their energy expenditure and substrate oxidation were measuredusing indirect calorimetry, and their appetite wasmeasured using visualanalog scales over a 4-h period. Blood samples were taken at 0, 90, and240 min postprandially. Food intake was measured using weighed foodrecords over the remaining 24-h period. Data were analyzed using 2-factor, crossover, repeated-measures ANOVA or 2-sample independentt test.

Results:NWandOWparticipants had decreased (P< 0.05) hunger,desire to eat, and cravings for a snack over the 4-h period followingO3FA consumption. O3FA consumption decreased calorie intake by∼13 kcal/d in NW participants and ∼43 kcal/d in OW participants.There was a significant time × diet interaction (P < 0.0001) forthermic effect of feeding and postprandial energy expenditure, withhigher values at 120 min following O3FA consumption (0.07 ± 0.02 vs.0.01± 0.02 kcal/min, P< 0.01) in both NW and OW (data combined).Carbohydrate oxidation was higher in OW participants, with no effectof O3FA. Fat oxidation was higher (P< 0.01) at 60 min following O3FAconsumption for both NW and OW participants. There was no effectof O3FA on postprandial triglyceride or glucose concentrations. O3FAdecreased postprandial plasma free fatty acids in both groups.


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Conclusions: This study indicates that O3FA consumption affectspostprandial responses in both NW andOW children. O3FA consump-tion increases the TEF and fat oxidation, and reduces hunger inNWandOW when compared with an isocaloric control.

This trial was registered at clinicaltrials.gov as NCT03139773.Funding SourcesArkansas Biosciences Institute. Beverages were provided by Smart-

fish (Oslo, Norway).

Raptor Deletion Impairs Chylomicron Production in HumanEpithelial Colorectal Caco-2 Cells (P10-049)

Bo He and Regis Moreau

University of Nebraska-Lincoln

Objective: The aim of this study was to investigate the role of themechanistic target of rapamycin complex 1 (mTORC1) in chylomicronproduction in Caco-2 cells.

Methods: Lentivirus-mediated shRNA technology was used toknock down the mTORC1 signaling-associated proteins Raptor andTSC2 to create cell lines that exhibit basal (shScramble), low (shRaptor),or high (shTSC2) mTORC1 activity. Cells were grown on plates todetermine 1) the proliferation rate, by counting cell numbers, and 2) thedifferentiation status by detecting alkaline phosphatase (ALP) activity.To stimulate chylomicron production, differentiated cells grown oninserts were incubatedwith a 1.2mMoleate-1.5%BSA complex for 16 h.

Results: Cell validation data showed Raptor protein to be lowerby 96% and p-p70S6K lower by 96% in shRaptor vs. shScramble cells(P < 0.05), and TSC2 protein lower by 90% and p-p70S6K higherby 86% in shTSC2 vs. shScramble cells (P < 0.05). We also foundthat mTORC1 altered Caco-2 cell proliferation and differentiation.Specifically, the proliferation rate of shTSC2 cells was 15% higher(P < 0.05) than that of shScramble cells, whereas no difference wasobserved between shRaptor and shScramble cells. During differentia-tion, shRaptor cells exhibited lower ALP activity than shScramble cells(–9.5%, P < 0.05), whereas shTSC2 exhibited higher ALP activity thanshScramble cells (+10.8%, P < 0.05). After incubation with oleate,we examined the expression of proteins and enzymes involved inchylomicron synthesis. Gene expression data showed that transcriptionof APOB (–87%), MTP (–94%), FASN (–76%), DGAT1 (–77%), andDGAT2 (–81%) was comprehensively repressed (P < 0.05) in shRaptorvs. shScramble cells. The abundance of 1) cellular apoB100 (–89%),MTP (–63%), and FASN (–80%), 2) media apoB100 (–51%), and 3)media apoB48 (–66%)were all significantly (P< 0.05) lower in shRaptorthan in shScramble cells. Moreover, Raptor knockdown decreasedmedia triacylglycerides by 69% (P < 0.05).

Conclusions: Our results reveal the critical role of intestinalmTORC1 in chylomicron metabolism. Intestinal mTORC1 signalingmay prove to be a therapeutic target against chylomicron overproduc-tion and associated hypertriglyceridemia.

Funding SourcesSupported by USDA Hatch Act and USDA-NIFA.

Nutrient Sensing andMetabolicChanges afterMethionineDepri-vation inPrimaryMuscleCells ofTurbot (ScophthalmusmaximusL.)(P10-050)

Gen He, Haowen Jiang, Fuyun Bian, Huihui Zhou, Xuan Wang,Kaidi Wang, and Kangsen Mai

Ocean University of China

Background: The low methionine content in plant-based diets isa major limiting factor for feed utilization by animals. However, themolecular consequences triggered by methionine deficiency have notbeen well characterized, especially in fish species, whose metabolism isunique in many aspects and important for aquaculture industry.

Objectives: The aim of this study was to characterized themolecular consequences triggered by methionine deficiency in turbot(Scophthalmus maximus L.).

Methods: In the present study, the primary muscle cells of turbotwere isolated and treatedwith or withoutmethionine for 12 h in culture.The responses of nutrient sensing pathways, the proteomic profiling ofmetabolic processes, and the expressions of key metabolic moleculeswere systematically examined.

Results: Methionine deprivation (MD) suppressed target of ra-pamycin signaling, and activated AMP-activated protein kinase andamino acid response pathways. Reduced cellular protein synthesis andincreased protein degradation by MD led to increased intracellularfree amino acid concentrations and degradations. MD also reducedglycolysis and lipogenesis while stimulating lipolysis, resulting in adecreased intracellular lipid pool. MD significantly enhanced energyexpenditure by stimulating the tricarboxylic acid cycle and oxidativephosphorylation.

Conclusions: Our results have identified a comprehensive set oftranscriptional, proteomic, and signaling responses generated by MDand provide molecular insight into the integration of cell homeostasisand metabolic controls in fish species.

Funding SourcesThis study was supported by grants from the National Natural

Science Foundation to GH (31,572,627), the 973 program to KM(2014CB138602), and the Aoshan Talents Cultivation Program to GH(2017ASTCP-OS12).


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The Influence of Noncaloric Artificial Sweetener Consumptionon Lipid and Energy Metabolism (P10-051)

Brian R Hoffmann and Martin Hessner

Medical College of Wisconsin

Background: The negative implications of consuming high dietarysugar have long been linked to systemic health problems. As theseconcerns have grown, dietary noncaloric artificial sweeteners have beenintroduced as an alternative to sugar; however, supporting scientificdata regarding the safety of these food additives is limited andcontroversial. It has also come to the forefront that interactions ofnoncaloric artificial sweeteners and an individual’s microbiome canalter metabolic processes.

Objective: This study aimed to improve our understanding ofnoncaloric artificial sweetener consumption by defining howmetabolicand microbiome alterations influence risk factors associated withmetabolic syndrome, obesity, and diabetes.

Methods: To better understand the metabolic consequences ofdietary consumption of noncaloric artificial sweeteners, we supple-mented the diet of diabetes-susceptible BioBreeding (BB-DR) ratswith aspartame and acesulfame potassium for 3 wk in their drinkingwater. A complementary group of BB-DR rats received bacitracinor streptomycin in addition to the sweeteners; we have previouslyshown this reverts the BB-DR microbiome to a healthier population.Weight, food intake, water intake, and blood glucose concentrationswere monitored throughout the study. At the end of the dietarysupplementation, mass spectrometry-based metabolomics analysis wasperformed on plasma samples from each group.

Results: At the end of the study, the BB-DR rats consumingthe artificial sweeteners had a significant increase in blood glucosein comparison with normal and glucose diets, despite consumingless food and water. Treatment of BB-DR rats consuming theseartificial sweeteners with bacitracin or streptomycin alleviated thiselevated blood glucose. Plasma metabolomics analysis revealed distinctalterations in lipid and energy metabolism in the rats consumingthe sweeteners, with a greater effect resulting from acesulfame potas-sium consumption. Specific decreases in the homolipidemic agentnicotinyl and tricarboxylic acid intermediates were detected, whereasincreases in indicators of muscle breakdown and triglycerides wereobserved.

Conclusions: This study suggests that aspartame and acesulfamepotassium contribute to metabolic derangements that may be anindicator of metabolic syndrome, obesity, and diabetes.

Funding SourcesSupport for this project has been provided by the NIH National In-

stitute of Diabetes and Digestive and Kidney Diseases (K01-DK105043to BRH), the Mayo Clinic Metabolomics Core via a Pilot Awardthrough grant U24DK100469, and the Department of BiomedicalEngineering at the Medical College of Wisconsin and MarquetteUniversity, Milwaukee, WI.

Watermelon Consumption Reduces Body Weight, Body MassIndex, and Blood Pressure in Overweight and Obese Adults (P10-052)

Mee Young Hong, Tiffany Lum, Amanda Marx, Shirin Hoosh-mand, Changqi Liu, and Mark Kern

San Diego State University, CA

Background: Watermelon consists of 92% water and is high infiber, vitamins, minerals, and bioactive phytochemicals. Althoughsome studies have demonstrated the beneficial effects of watermelonsupplementation on metabolic diseases, no study has explored wa-termelon consumption’s potential mechanism relating to body weightmanagement. Furthermore, those prior studies used watermelon juice,powder, or extract as their treatment rather than fresh watermelon.

Objective: The objective of this study was to determine the effect ofdaily consumption of 2 cups of fresh watermelon on body weight (BW),body mass index (BMI), blood pressure (BP), and waist-to-hip (W/H)ratio in overweight and obese adults.

Methods:A crossover design was used to evaluate BW, BMI, systolicblood pressure (SBP), diastolic blood pressure (DBP), and W/H ratiomeasurements in 33 overweight or obese adults (20 male, 13 female;age: 26.5 ± 13.3 y; BMI: 30.6 ± 5.2 kg/m2) at baseline and after4 wk. Subjects received 2 isocaloric snacks (92 kcal per serving) offresh watermelon and reduced fat cookies. All subjects were assignedto 4 wk of watermelon consumption (2 cups) followed by a 2- to 4-wk washout period, then 4 wk of reduced fat cookie consumption(6 cookies).

Results: BW and BMI decreased significantly after watermelon con-sumption compared with reduced fat cookie consumption (P = 0.006).A significant decrease in SBP was found following watermelonconsumption, whereas significant increases in SBP andDBPwere foundfollowing cookie consumption (P < 0.05). Furthermore, reductions inwaist measurements (P = 0.063) and W/H ratio (P = 0.052) trendedtoward significance after the watermelon phase.


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Conclusions: Management of body weight through dietary meansis beneficial for the prevention of chronic diseases. This study showsthat reductions in BW, BMI, and BP can be achieved through dailyconsumption of watermelon, thus reducing risk factors associatedwith overweight and obesity. Further studies on potential biomark-ers and mechanisms of action responsible for these results arewarranted.

Funding SourcesThe National Watermelon Promotion Board.

Effect of Snack Type on Satiety in an Acute Feeding Study ofOverweight and Obese Subjects (P10-053)

Mee Young Hong, Traci Roberts, Natasha Godwin, ShirinHooshmand, and Mark Kern

San Diego State University, CA

Background: Snacking can be an important component of weightmanagement, and snack choice is made based on a variety of factors,such asmacronutrient composition, satiety effects, and total calories perserving.

Objective: The aim of this study was to examine the effects on bloodsugar and satiety in an isocaloric portion of 2 commonly consumedsnacks: mixed nuts and pretzels. We hypothesized that mixed nutswould promote reduced hunger and beneficial metabolic responses,which would improve weight loss.

Methods: Subjects (n = 54; age: 29.7 ± 1.9 y; body mass index:31.5 ± 0.6 kg/m2) came to the testing laboratory in the morningafter an overnight fast, and an antecubital blood sample was taken forbaseline measures of glucose and appetite hormones. A baseline satietyquestionnaire was also given. Subjects were then instructed to consumeeither the mixed nuts (42 g) or pretzels (69 g), along with 16 oz ofwater in 15 min. Satiety responses were measured at 20, 40, 60, 90, and120 min, and a second blood sample was taken at 60 min to measureblood glucose.

Results: Both mixed nuts and pretzels significantly increasedmeasures of satiety (P < 0.001), with greater satiety with the pretzelsthan with the mixed nuts (P < 0.001). Pretzel consumption alsoshowed a significant increase in postprandial blood glucose (P< 0.001)compared with baseline, whereas nut consumption did not significantlyincrease postprandial blood glucose concentrations. The results alsoshowed an inverse correlation of postprandial glucose and hungerfeeling (coefficient = –0.274, P = 0.049), and a positive correlationwith fullness (coefficient = 0.441, P = 0.001) at 60 min after snackconsumption.

Conclusions: A possible explanation for the increased satietyresponses of pretzels over mixed nuts is the glucostatic theory, whichstates that changes in arteriovenous glucose differences are detectedby glucoreceptors, which affect energy intake. Therefore, increases inblood glucose concentrations result in increased feelings of satiety; a

decrease in blood glucose has the opposite effect. It is unclear whetherthese increased feelings of satiety would reduce subsequent energyintake at the next meal. This warrants further investigation into howsnacking affects weight management.

Funding SourcesAmerican Heart Association (16GRNT31360007).

Estimating Gestational Weight Gain Change Based on Increasesin Fruit Intake: A Modeling Analysis (P10-054)

Amber Hromi-Fiedler,1 Pamela Clark,2 Grace Damio,3 SofiaSegura-Pérez,3 Grace Kollannoor Samuel,4 Angela Bermúdez-Millán,5 and Rafael Pérez-Escamilla1

1Yale University, CT; 2Hartford Hospital, CT; 3Hispanic HealthCouncil; 4State of Minnesota; and 5University of Connecticut HealthCenter

Objective: The aim of this study was to estimate changes ingestational weight gain (GWG) by lowering energy density throughincreased fruit intake.

Methods: A total of 74 overweight and obese pregnant Latinascompleted a prenatal 24-h recall in a mixed methods study designed toinform an intervention to improve prenatal fruit and vegetable intake.Women were included if they were: 1) ≥18 y old; 2) in their secondtrimester of pregnancy; 3) on a low income; 4) Latina; 5) having a single-ton pregnancy; 6) overweight or obese (i.e., pregravid body mass index≥25); and 7) not on a restricted diet. Fruit servings were computedwith the Nutrient Data System for Research software. GWG changeassociated with energy density changes related to incremental increasesin fruit intake was predicted using multivariable linear regression.

Results: Energy density (ED) was 1.27 kcal/g and median fruitintakes were an average of 1.1 servings. Computer simulations foundthat increasing fruit intake incrementally by 0.5 servings (e.g., 0.5of an apple with skin) decreased the ED by 0.84 kcal/g, 1 servingdecreased the ED by 1.36 kcal/g, 1.5 servings decreased the ED by1.84 kcal/g, 2 servings of fruit decreased the ED by 0.15 kcal/g, and3 servings decreased the ED by 0.20 kcal/g. Changes in ED acrossall scenarios improved fiber, water, folate, vitamin C, carotenoids,and iron intakes. Reducing prenatal ED by 0.15 kcal/g (i.e., 2 fruitservings) was estimated to reduce GWG by 2.51 lbs. Similarly, reducingprenatal ED by 0.20 kcal/g (i.e., 3 fruit servings) was estimated toreduce GWG by 3.35 lbs. Subsequently, reducing prenatal ED by 0.20kcal/g resulted in a decrease of excessive GWG prevalence by 11.8% inour sample.

Conclusions: Preliminary findings suggest that increasing prenatalfruit intake has the potential to modify energy density, subsequentlyreducing gestational weight gain among overweight and obese pregnantLatinas.

Funding SourcesNIH/NINR (Grant number: 1R21NR013970-01).


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Evenly Distributing Daily Protein Intake Does Not DifferentiallyInfluence Improvements in Cardiometabolic Health Induced byWeight Loss and Resistance Training (P10-055)

Joshua L Hudson,1 Jung Eun Kim,2 Douglas Paddon-Jones,3 andWayne W Campbell1

1Purdue University, IN; 2National University of Singapore; and3University of Texas Medical Branch

Objectives: We sought to assess the effects of within-day proteinintake distribution on changes in clinical fasting cardiometabolic healthindicators and 24-h glucose control after 16 wk of dietary energyrestriction and resistance training.

Methods: With the use of a randomized, parallel-design, 41 menand women (mean ± SEM: age: 35 ± 2 y; body mass index: 31.5 ± 0.5kg/m2) performed resistance-type training 3 d/wk and consumed acontrolled diet containing 750 kcal/d less than their estimated energyrequirement for 16 wk. Subjects consumed 90 g protein/d (∼1.0 g ·kg–1 · d–1) in either a skewed (10 g at breakfast, 20 g at lunch, 60 g atdinner; n = 20) or even (30 g at breakfast, lunch, and dinner; n = 21)distribution pattern. Fasting clinical cardiometabolic health indexes and24-h continuous interstitial glucose concentrations were measured pre-and postintervention. Glucose and insulin responses to a 12-h mealglucose tolerance test (12-h MGTT) were measured postintervention.

Results: Within-day protein distribution did not affect any car-diometabolic health or glucose control outcomes. Among all subjects,over time, body mass decreased by ∼8% and whole-body strengthincreased by ∼20%. Fasting serum glucose (least-squares mean ± SE;–2.9 ± 1.6 mg/dL), insulin (–5.5 ± 1.2 µIU/mL), total cholesterol(–21 ± 4 mg/dL), HDL (–3 ± 1 mg/dL), LDL (–13 ± 3 mg/dL),and triglyceride (–26 ± 7 mg/dL) concentrations all decreased (maineffect of time, P < 0.05). The 24-h continuous interstitial glucoseconcentration average (–7.6± 2.6mg/dL), variability (–2.7± 1mg/dL),peak (–14 ± 4 mg/dL), fluctuation (peak – minimum; –13 ± 3 mg/dL),and time spent above 100 mg/dL (–242 ± 87 min/d) also decreased(main effects of time, P < 0.05). There was no effect of proteindistribution on postintervention 24-h continuous interstitial glucoseoutcomes, 12-h MGTT glucose, and insulin positive incremental areasunder the curve.

Conclusions: The effectiveness of dietary energy restriction com-bined with resistance training to improve clinical indices of car-diometabolic health and daily glucose control is not influenced by thewithin-day distribution of protein.

Funding SourcesPorkCheckoff;American EggBoard-EggNutritionCenter; National

Dairy Council; Beef Checkoff; and the NIH, Indiana Clinical andTranslational Sciences Institute, Clinical Research Center, Grant #UL1TR001108.

Liver Protein Synthesis Increases in a Rat Model of Diet-InducedObesity and Metabolic Syndrome (P10-056)

Jean-Francois Huneau,1 Olivier Mantha,2 Véronique Mathé,2Dominique Hermier,2 Francois Mariotti,2 and Hélène Fouillet2

1AgroParisTech; and 2UMR PNCA

Objective: Little is known regarding the effect of obesity andinsulin resistance on protein metabolism. The aim of this study was to

explore the links between the degree of obesity and insulin resistanceand protein synthesis in several tissues, as shown by the phenotypicvariability of rats exposed to a high-fat diet.

Methods: A total of 42 male Wistar rats were fed a high-fat dietfor 4 mo and then killed for tissue sampling. Weight, total and visceraladiposities, homeostatic model assessment insulin resistance index,liver, and plasma triglycerides were measured to compute an obesityand metabolic syndrome score (ZOMS) based on the z scores of theseparameters. Ratswere injected intraperitoneallywithD2O (20 g/kg, 99.9atom%) 2 d before being killed, and given free access to D2O (5 atom%)in their drinking water until their death. In the sampled tissues, D-enrichment of free and protein-bound alanine was measured by GC-MS and used to calculate fractional (FSR) and absolute (ASR) proteinsynthesis rates using a precursor-product model.

Results: After 4 mo of high-fat feeding, there was a continuumof phenotypes, from lean and insulin-sensitive to obese and insulin-resistant rats, with a range of variations in ZOMS parameters (given asmean [range] in Table 1).

Regression analyses indicated significant positive relations betweenZOMS and protein FSR in the plasma (β = 0.074 ± 0.024) and to alesser extent in the gastrocnemius muscle (β = 0.004 ± 0.001). Positiverelations between ZOMS and protein ASR were strong in the liver(β = 13.80 ± 1.82), plasma (β = 17.11 ± 6.25), and kidney (β = 3.68± 0.81), but weak in the gastrocnemius muscle (β = 0.65 ± 0.24). Norelations between protein synthesis rates and ZOMS were observed inthe other tissues (heart, tibialis muscle, small intestine).

Conclusion: In the liver and to a lesser extent a skeletal muscle,protein synthesis increased with the degree of obesity and metabolicsyndrome. This is probably because our study encompassed the earlyphase of obesity development, where the gain in fatmass is accompaniedby a gain in lean mass owing to a higher anabolic orientation of both fatand protein metabolisms.

Funding SourcesFunded by INRA, department AlimH.

TABLE 1

DietaryProtein Intake andLevodopaManagement inParkinson’sDisease (P10-057)

LeahMHurley,Dara Lobuono, LeslieMahler, and Ingrid Lofgren

University of Rhode Island

Objective: Parkinson’s disease (PD) is a progressive neurodegener-ative movement disorder that can cause resting tremors, slowness ofmovement, cognitive decline, difficulty swallowing, and gastrointestinalissues, all of which can negatively impact dietary intake. Levodopa is themost commonmedication to improvemotor functioning and quality oflife, and to reducemorbidity andmortality in persons with PD (PwPD).


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However, levodopa and dietary protein compete for absorption in thegut and across the blood-brain barrier. Dietary proteins are essentialfor muscle mass retention, growth, and physical functioning, whichgenerally deteriorate with PD progression; 0.8 g/kg is recommended.The objective of this cross-sectional study was to describe the dietaryprotein intake in relation to levodopa intake in PwPD.

Methods: Dietary recalls (2 d) were collected from 7 PwPD usingthemultiple pass method andNutrition Data System for Research 2017.Pearson’s correlations determined the strength of any correlations.

Results: Participants consumed a mean of 2096 ± 633 kcal and67 ± 23.5 g protein over the course of 4.5 ± 1.3 meals/d. Meanprotein intake ranged from 0.7 to 1.2 g · kg–1 · d–1, with a mean of44.2 ± 13.9 g/d of animal and 31.2 ± 11.2 g/d of plant proteins. Themost commonprotein sources wereGreek yogurt, chicken breasts, eggs,oatmeal, peanuts, and cereal grains. The average time on levodopa was4.7± 2.8 y and the mean number of doses was 3.9± 1.2 doses/d. Therewere strong positive correlations (P< 0.01) between total plant proteinsand total calorie intake (r= 0.927), and between total plant proteins andbody mass index (r = 0.894). There were strong negative correlationsbetween years since diagnosis and animal protein intake (r = –0.876;P < 0.01) and total protein intake (r = –0.852; P < 0.05), with a largenegative correlation (r= –0.624) with grams of protein per kilogram ofbody weight.

Conclusions: Our results show that protein choices and overallintake may change as PD progresses; longitudinal studies are needed tofurther explore this pattern. People with PD may need more educationabout the food-drug interaction as time on levodopa increases.Therefore, dietary protein monitoring in PwPDmay be warranted, andreferrals to registered dietitians are appropriate.

Funding SourcesN/A.

Dietary Saturated Medium-Chain and Polyunsaturated Long-Chain Fats Decrease Circulating Gut Microbiome Postbiotics andInflammatory Cytokines in Cats (P10-058)

Matthew I Jackson and Dennis Jewell

Hill’s Pet Nutrition

Objective: Dietary fats are mostly absorbed in the upper gastroin-testinal tract, but consumption alters hindgutmicrobiome composition.The objective of this study was to determine the impact of a combi-nation of saturated medium-chain (sMCT) and polyunsaturated long-chain triglycerides (pLCT) on serum concentrations of gut microbialmetabolic products (postbiotics) and inflammatory cytokines in cats.Postbiotics can influence inflammatory status, with implications forhealth and disease.

Methods: We used a randomized 2 × 2 design (±sMCT, ±pLCT),with 4 groups, n = 16 per group. Cats were fed for 28 d and grouphoused in environments promoting social interaction, having accessto natural light that varied with season and opportunity to exercise.Health was confirmed through clinical indices. Collections were madeaccording to IACUC-approved protocols. Cats were fed to maintain ahealthy body weight. Serum gut microbiome postbiotics were assessedby metabolomics LC-MS. Inflammatory cytokines were measured byELISA from whole blood samples cultured in vitro with or withoutlipopolysaccharide (LPS) as a proxy for metabolic endotoxemia.

Results: Multivariate tests by postbiotic class showed thatsMCT + pLCT together, but not alone, decreased serum indole andphenol postbiotics, as well as sulfated host cometabolites. Univariatetests exposed drivers; for example, the putrefaction products 4-ethylphenol sulfate, 4-vinylphenol sulfate, and indolelactate were allsignificantly decreased by the combination of fats. The combinationof sMCT + pLCT also decreased cytokines in whole blood culturesfrom fed cats: FAS, granulocyte-macrophage colony-stimulating factor(GM-CSF), interleukin (IL)-2, IL-13, IL-8, and stromal cell-derivedfactor 1 (SDF-1) were decreased 15–64% relative to the additive effectsof both fats separately in cultures without added LPS. In cultures withadded LPS, the cytokines FAS, GM-CSF, IL-2, IL-13, SDF-1, and tumornecrosis factor α were decreased 10–90% relative to the additive effectsof both fats separately.

Conclusions: The combination of sMCT and pLCT decreasescirculating concentrations of feline gut microbial postbiotics, someof which are uremic toxins, and also decreases concentrations ofinflammatory cytokines in whole blood cultures from fed cats,regardless of culture stimulation by LPS. Dietary sMCT and pLCTfats in combination may improve functional dysbiosis and decreaseinflammation in cats over 4 wk.

Funding SourcesHill’s Pet Nutrition, Inc., Topeka, KS.

Effect of Partial Substitution of Wheat Flour with a Flour Blendof Okra, Lentils, Barley, and Fenugreek on the Glycemic Index ofCommonly Consumed Starchy Foods (P10-059)

Alexandra L Jenkins,1 Nadja Piatka,2 Adish Ezatagha,1 and RajenManicka2

1GI Labs; and 2Holista Foods

Objective: A blend consisting of okra, lentils, barley, and fenugreekwas developed with the aim to reduce the rate of glucose absorptionfrom flour used in processed starchy foods. The objective of this studywas to measure the glycemic index (GI) of a range of commonlyconsumed starchy foods in which wheat flour was partially substitutedwith the plant-based blend.

Methods: Four different starchy foods containing the blend(HolistaTM GILiTE) were tested: a muffin, a breakfast cookie, noo-dles, and bread. The GIs were determined using the standard ISO26,642:2010 method for GI determination. Healthy male or femalesubjects aged 18–75 y with a body mass index (BMI) <35 kg/m2 wererecruited. Each test meal contained 25 or 50 g of available carbohydrate.Capillary blood samples for blood glucose analysis were taken at –5, 0,15, 30, 45, 60, 90, and 120 min. Each subject consumed the test foodonce and the reference food at least twice to allow calculation of the GI.Palatability was assessed using a visual analog scale.

Results: A total of 23 participants were recruited (12 male, 11female; age: 45 ± 14 y; BMI: 28.3 ± 5.5 kg/m2). Each food wastested by 10 participants, and each participant consumed ≥1 test foodand 2 reference foods at GI Labs. The GIs of the muffin, cookie,noodles, and bread were 48 ± 2, 52 ± 3, 38 ± 4, and 66 ± 8,respectively. All foods were well tolerated. The palatability scores werenot statistically significantly different from the white bread control butwere significantly higher than the dextrose control (P < 0.05).


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Conclusions: Partial substitution of wheat flour with a flour blend ofokra, barley, lentils, and fenugreek resulted in 3 of the products fallinginto the low GI category; the exception was the bread, which fell intothe medium GI category. Further research needs to be undertaken todetermine the extent of the reduction in GI achieved by the blend.

Funding SourcesHolista Colltech Ltd and Holista Foods.

Effects of Dried Chokeberry (Aronia melanocarpa) on MetabolicDisorder in C57BL/5J Mice Fed a High-Fat and High-Fructose Diet(P10-060)

Oeuk Jeong and Hyun-Sook Kim

Sookmyung Women’s University, South Korea

Background:Ahigh-fat and high-fructose (HFHF) diet is known tocause metabolic syndrome in animals. As the intake of the HFHF dietincreases, the incidence of chronic diseases such as obesity, metabolicsyndrome (MetS), and type 2 diabetes, increases.

Objective: The aim of this study was to investigate the effects ofdietary dried chokeberry powder in an HFHF diet–induced metabolicdisorder mouse model.

Methods: After 1 wk of adaptation, 6-wk-old male C57BL/6J micewere randomly divided into 3 groups (n = 7 for each group): a normaldiet group (ND), an HFHF diet group (HF), and an HFHF diet with1% chokeberry group (CH). The mice in each group were fed withcorresponding diets for 10 wk.

Results: The body weight gained by the mice over 10 wk was6.38 ± 1.66 g in the ND group, 17.99 ± 2.87 g in the HF group,

and 9.84 ± 2.52 g in the CH group. The liver weight in the HFgroup (1.18 ± 0.10 g) was significantly higher than in the ND group(0.96 ± 0.10 g), and the liver weight of the CH group was significantlylower than that of the HF group (P = 0.0003). The abdominalfat weight in the CH group was lower than that in the HF group(P < 0.0001), and the epididymal fat weight in the CH group was alsosignificantly lower than that in the HF group (P < 0.0001). There weresignificant decreases in serum insulin concentration and homeostaticmodel assessment insulin resistance index (HOMA-IR) (P = 0.0006,P= 0.0003, respectively). Moreover, the endpoint fasting blood glucoseconcentration was decreased in the CH group compared with the HFgroup (P= 0.0222). Serum triglyceride and total cholesterol concentra-tion were decreased in the chokeberry-fed group compared with theHFgroup (P < 0.0001). The concentrations of serum cholesterols changedwith the intake of chokeberry. In the CH group, HDL cholesterol tendedto be lower than in the HF group (P = 0.0707). However, LDL, VLDL,and non-HDL cholesterols were significantly lower than in the HFgroup (P < 0.0001). As a result of oral glucose tolerance tests, theCH group was shown to have lower blood glucose concentrations at90 and 120 min than the HF group, though the difference was notsignificant. Hepatic superoxide dismutase 2 and glutathione peroxidaseexpression did not differ between the groups (P = 0.3295, P = 0.2153,respectively).

Conclusion: These results suggest that dietary chokeberry powdersupplementation may exert a protective effect by improving the serumlipid concentration through the control of insulin resistance in HFHFdiet–induced metabolic disorder mice.


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FIGURE 1 Body weight change.The different letters (a,b,c) within a column indicate significant differences (P < 0.05) determined by Duncan’s multiple range test.-ns: not significant

FIGURE 2 Insulin and HOMA-IR concentrations.The different letters (a,b) within a column indicate significant differences (P < 0.05) determined by Duncan’s multiple range test.-ns: not significant


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FIGURE 3 Concentrations of serum cholesterols.The different letters (a,b) within a column indicate significant differences (P < 0.05) determined by Duncan’s multiple range test.-ns: not significant

FIGURE 4 Experimental scheme.


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FIGURE 5 Oral glucose tolerance tests.The different letters (a,b) within a column indicate significant differences (P < 0.05) determined by Duncan’s multiple range test.-ns: not significant

FIGURE 6 Hepatic protein expressions.The different letters (a,b) within a column indicate significant differences (P < 0.05) determined by Duncan’s multiple range test.-ns: not significant


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Effects of Medium-Chain Diacylglycerol and Equivalent Intakesof Long-Chain Triacylglycerol Oils as Part of Muffins on Postpran-dial Lipid Metabolism (P10-061)

Minjoo Kim,Minkyung Kim, Hye Jin Yoo, Ayoung Lee, and JongHo Lee

Yonsei University, South Korea

Objective: The aim of this study was to assess the ability ofmedium-chain diacylglycerol (MCDG) oils to lower concentrations ofpostprandial triglycerides (TGs) and plasma fatty acids.

Methods: This double-blind, randomized, crossover trial evaluatedeffects ofMCDGoils on the postprandialmetabolic responses in healthymen and women with normal weight (n = 28). Each subject carriedout a meal tolerance test twice during the study period, with a 7-dinterval. Muffins that contained 15 g of MCDG oils (MCDG muffins,test products) or long-chain triacylglycerol (LCT) oils (LCT muffins,placebo products) were used for the meal tolerance test. Blood sampleswere collected in the fasting condition (0 h) and at 2, 3, 4, and 6 h afteringestion of the muffins.

Results: Postprandial TG concentrations were significantly lowerat 3 and 4 h after consuming the MCDG muffins compared withthe LCT muffins. Postprandial chylomicron-TG concentrations weresignificantly lower at 4 h after consuming theMCDGmuffins comparedwith the LCT muffins. The area under the curve of postprandial TGand chylomicron-TG decreased by ∼13% when the subjects ingestedMCDG muffins rather than LCT muffins. Postprandial concentrationsof plasma fatty acids, including palmitic acid, stearic acid, and linoleicacid, were decreased after MCDG muffin intake compared with LCTmuffin intake.

Conclusion: These results indicate that dietary MCDG oils de-creased postprandial TGs, chylomicron-TGs, and plasma fatty acidscompared with dietary LCT oils. This might be because of metaboliccharacteristics of MCDG oil in the digestive tract.

Funding SourcesThis study was supported by the Bio-Synergy Research Project

(NRF-2012M3A9C4048762), the Mid-Career Researcher Program(NRF-2016R1A2B4011662), and the Basic Science Research Program(NRF-2017R1C1B2007195) through the National Research Foundationof Korea (NRF) funded by the Ministry of Science, ICT & FuturePlanning.

The Synergistic Contribution of Lactobacillus johnsonii N6.2 andPhytophenols in Reducing Metabolic Syndrome via the MechanisticTarget of Rapamycin Pathway (P10-062)

Danielle N Kling, Evon DeBose-Scarlett, Leandro D Teixeira,Graciela L Lorca, and Claudio F Gonzalez

University of Florida

Background: Metabolic syndrome (MetS) is a precursor to someprevalent diseases, like type 2 diabetes and obesity. It is diagnosedbased on having 3 of the following criteria: abdominal obesity, hy-pertriglyceridemia, low HDL, high blood pressure, and hyperglycemia.The mechanistic target of rapamycin (mTOR) is a regulator of insulinsignaling, adipogenesis, and glucose metabolism. Regulation of mTORis essential for metabolic homeostasis. Research into a beneficial sym-biont, Lactobacillus johnsonii N6.2, has revealed 2 key characteristics

that may aid in modulating mTOR: it is negatively correlated withdiabetes development, and it encodes for esterases responsible forcleaving phytophenols from dietary fiber. This is important as manyinhibitors of the mTOR pathway have phenolic constituents.

Objectives: The aims of this study were to analyze the effectivenessof a high-fat diet (HFD) in evoking MetS, then use a probiotic strainin concert with natural food components to reduce mTOR-mediatedmetabolic disease onset.

Methods: Rats were fed an HFD or a reduced-energy diet (RED)and were treated 3 times a week with blueberry extract and/or L.johnsonii for 16 wk. During this time, weight and triglyceride andglucose concentrationsweremonitored.mTORpathway expressionwasanalyzed via Western blot by evaluating activating phosphorylationsof upstream and downstream biomarkers. The significance of theseactivations was determined by analyzing expression levels of mTOR-induced genes. mTOR stimulatory signals, such as insulin, tryptophan,and inflammatory cytokines, were analyzed by ELISA, LC-MS/MS, orquantitative polymerase chain reaction.

Results: Males were more susceptible to HFD effects than females,as noted through increases in weight and triglyceride and glucose con-centrations. Females, but more notably males, experienced increasedliver fat deposits. Treatments were more effective in reducing mTOR-activating phosphorylations and downstream gene expression in HFD-fed females and RED-fed males. Insulin, tryptophan, and inflammatorycytokines did not show any significant differences between groups.

Conclusions: Differences in MetS markers may be gender related,as males were susceptible to HFD-inducedMetS. Treatment effects maybe affected by gender or MetS status, as treated HFD females and REDmales experienced a decrease in mTOR expression.

Funding SourcesSupported by the National Institute of Food and Agriculture USDA

Grant No. 2015-67,017-23,182 to GLL and CFG.

FattyAcidsUnique toDairy Fat ImproveGlucoseTolerance (P10-063)

Jana Kraft, Helen Walsh, Tarleton Emily, Bunn Janice, and KienCraig

University of Vermont

Background: Observational studies have identified a positive roleof dairy fat in the prevention of type 2 diabetes. The principalcomponent(s) responsible are yet to be determined.

Objectives: Our objective was to test the hypothesis that uniquefatty acids (FAs) found in dairy fat per se and derived from rumenmicrobial synthesis would alter glucose homeostasis and be protectivewith respect to glucose intolerance.

Methods: We conducted a clinical trial with 21 healthy adults [10women, mean ± SEM body mass index (BMI): 21.3 ± 0.7 kg/m2; 11men, BMI: 23.8± 0.9 kg/m2]. A double-masked, randomized crossoverdesign was employed with 2 sequential 3-wk dietary interventionspreceded by a 1-wk run-in period (reflecting a representative US diet)to compare a eucaloric balanced DASH-style diet containing either 3daily servings of whole yogurt (dairy fat diet; DF) or fat-free yogurtsupplemented with a control fat (control diet). At the end of each aperiod, the FA composition of serum phospholipids was assessed, anda frequently sampled intravenous glucose tolerance test was performed


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to quantify the effects of diet on glucose uptake, insulin secretion, andinsulin sensitivity.

Results: The FA composition of serum phospholipids, a markerof dietary compliance and intake, reflected the FA composition ofthe respective diet. During the DF diet, there was a higher contentof branched-chain FAs, total trans-18:1, 18:1 t11 (vaccenic acid),and 18:2 c9, t11(rumenic acid) in serum phospholipids (P < 0.001).Consumption of 3 daily servings of whole yogurt (DF) resulted in a26% higher glucose effectiveness (SG; index of insulin independentglucose uptake; P = 0.014) and a borderline higher (12%) acute insulinresponse to glucose (AIRG; index reflecting β-cell sensitivity to glucose;P = 0.053).

Conclusions:Regular intake of dairy fat promotes improved glucosetolerance by stimulating both insulin-independent glucose uptake andenhanced insulin secretion. Our results show that even modest intakeof dairy fat may be necessary to maintain normal glucose homeostasis.

Funding SourcesNational Dairy Council (NDC).

Biomarkers of Caloric Restriction Predict Nonalcoholic FattyLiver Disease in the Multiethnic Cohort—Adiposity PhenotypeStudy (MEC-APS) (P10-064)

Bruce Kristal,1,2 Marina Belenky,1,2 Defne Altan,1,2 Matthew JSniatynski,1,2 Vasant Marur,1,2 Rose Gathungu,1,2 Unhee Lim,3Kris Monroe,5 Thomas Ernst,4 John Shepherd,6 Steve Buchthal,4Bo Fan,6 Iona Cheng,7 Meredith Hullar,8 Johanna Lampe,8Timothy Randolph,8 Lynne Wilkens,3 and Loic Le Marchand3

1Harvard Medical School, MA; 2Brigham and Women’s Hospital,MA; 3University of Hawaii Cancer Center and 4John A Burns School ofMedicine, University of Hawaii; 5Keck School of Medicine, Universityof Southern California; 6School of Medicine, University of CaliforniaSan Francisco; 7Cancer Prevention Institute of California; and 8FredHutchinson Cancer Research Center, WA

Objectives: Caloric restriction (CR) is the most potent, robust,and reproducible known means of extending longevity and decreasingmorbidity in laboratory rodents. Despite 80+ y of research, therelevance of this observation for humans remains unknown, but it issupported by the established link between obesity and morbidity inhumans.

Objectives: The aim of the Multiethnic Cohort—Adiposity Pheno-type Study (MEC-APS) is to investigate metabolites associated withnonalcoholic fatty liver disease (NAFLD)—the most common cause ofchronic liver disease worldwide—across 5 major US racial and ethnicgroups, which vary in NAFLD frequency by 2.5-fold. Although NAFLDis associated with obesity, prediction of NAFLD by total adiposity,including proxies such as weight and body mass index (BMI), is notstrong and differs across race and ethnic groups.

Methods: The MEC has followed 215,000 Hawaii and Los Angelesresidents of African, Japanese, European, Latino, and Native Hawaiianancestry since 1993–1996. In the MEC-APS, we measured liver fat bythe use of abdominal MRI and total body fatness by dual-energy X-ray absorptiometry in a cross-sectional subset of 1781 healthy womenand men aged 60–77 y with BMIs of 17–46 kg/m2. Redox active plasmametabolites were assessed using HPLC separations with coulometric

electrode array detection. For statistical analysis, we used the Mann-Whitney U test, regression analysis, and area under the receiveroperating characteristic curve (AUROC).

Results: Of 16 metabolites statistically associated with NAFLD, 7were previously linked to CR in studies of ad libitum fed and CRFBFN1 rats (males and females, different ages, and different extents anddurations of CR). Strikingly, the strongest 3 NAFLD markers (p-7 inboth sexes) were all previously CR-linked; all 3 are endogenous, verylow concentration in plasma, structurally unidentified molecules thatare followed using chromatographic and redox signatures. AUROCsfor NAFLD (∼0.75 in women, ∼0.85 in men) are comparable withthe best available markers. AUROCs are essentially equal across liverfat cut-offs (4.0–6.5%) near the standard clinical threshold for NAFLD(>5.5%), suggesting robustness and the potential to recognize at-riskindividuals. Limiting analysis to individuals who meet NAFLD alcoholconsumption criteria strengthened the associations, suggesting that themarkers capture themetabolic milieu specific to NAFLD, not overt fattyliver disease itself.

Conclusions: Biomarkers of CR in rats predict NAFLD men andwomen from 5 ethnic groups in the MEC-APS.

Funding SourcesP01 CA168530, U01 CA164973.

Basal Energy Expenditure in Long-Term Bariatric Patients:ComparisonbetweenPredictiveEquations and IndirectCalorimetryResults (P10-065)

Fernando Lamarca,1 Isabela Nogueira Martins Sena Rios,2Wilson José de Campos Filho,2 Gabriela Menezes PawlakPedroso,2 Nathalia Pizato,2 Eliane Said Dutra,1 and Kênia MaraBaiocchi de Carvalho1

1Postgraduate Program in Human Nutrition, University of Brasília(PPGNH/UnB) and Undergraduate Nutrition Course, UniversityCenter Unieuro (UNIEURO). Brasília, Federal District, Brazil; and2University of Brasília (UnB), Brasília, Federal District, Brazil

Objectives: To compare predictive equations (PEs) with indirectcalorimetry (IC) to estimate basal energy expenditure (BEE) in patientsin the late postoperative period after bariatric surgery.

Methods: We undertook an observational cross-sectional study, inwhich adult individuals (18–60 y old) of both sexes were included,who underwent bariatric surgery (a Roux-en-Y gastric bypass) 2–7 yago. Measured BEE was obtained by IC (Invoice Vmax29) after 8 h offasting for 30 min. The estimated BEE was obtained from the PEs ofHarris & Benedict (HB), Schofield, FAO/WHO/UNU (WHO), Mifflin-St Jeor, and Orozco-Ruiz. Body composition was assessed by dual-energy X-ray absorptiometry (Lunnar, DPX-IQ). Underestimation andoverestimation were considered when the estimated BEE differed fromthe valuemeasured by IC by±10%. The agreement of PEwere evaluatedthrough the intraclass correlation coefficient, with a 95% CI. Pearson’scorrelation was applied to investigate the association between the meanerror of the PE and anthropometric and body composition variables.

Results: A total of 22 participants (38.4 ± 6.4 y old; 86.4% women;body mass index: 29.9 ± 4.5 kg/m²) were enrolled in this analysis witha postoperative time of 3.8 y (95% CI: 2.6, 5.3). The mean measuredBEE value of 1479 ± 149 kcal, was significantly lower than the valuesobtained from the PEs of HB (1606 ± 239 kcal, P = 0.001), Schofield


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(1571 ± 232 kcal, P = 0.006), WHO (1595 ± 233 kcal, P = 0.001), andOrozco-Ruiz (1606 ± 247 kcal, P = 0.001), but not significantly lowerthan that obtained from the Mifflin-St Jeor equation (1514 ± 227 kcal,P = 0.26). However, the mean overestimations of the PEs comparedwith the IC were 8.5% for HB, 6.1% for Schofield, 7.7% for WHO, and8.4% for Orozco-Ruiz, which are all considered acceptable. TheMifflin-St Jeor equation presented the best agreement (r = 0.84, 95% CI: 0.62,0.93) to the value measured by IC. Positive associations between all themeans of the PE errors with BMI (r = 0.65, P = 0.001) and fat mass(r = 0.73; P < 0.001) were observed.

Conclusions: Although all the equations tested showed acceptablecomparison to the IC, the Mifflin-St Jeor equation presented the bestresults for the estimation of BEE in long-term bariatric patients. Theassociations of PE errors suggested that the greater the mass and bodyfat, the greater the disagreements of the results, whichmay compromisethe results in severely obese individuals.

Funding SourcesFoundation for Research Support of the Federal District (FAPDF)—

0193.001.462/2016.

Antiobesity Effects of Chestnut StarchModified byAmylosucrasefromDeinococcus geothermalis (P10-066)

Byung-Hoo Lee,1 Hansol Shin,1 Moon-Gi Hong,1 Eun-SookLee,2 and Dong-Ho Seo2

1Gachon University, South Korea; and 2Korea Food ResearchInstitute

Objectives: The primary objective of this study was to investi-gate whether modification of chestnut starch by amylosucrase fromDeinococcus geothermalis (DGAS) increases the resistant starch (RS)content and regulates obesity. Furthermore, we aimed to elucidate themechanism that prevents fat accumulation by prebiotic activity of RSfrom chestnut starch.

Methods:Chestnut starch was enzymatically modified by DGAS. Invitro analyses, including Englyst’s assay and determination of the starchdigestive pattern by mammalian mucosal α-glucosidases, were used todetermine the ratio of slowly digestible starch and RS in enzymaticallymodified chestnut starch. In an in vivo study, a high-fat diet (45% kcalfrom fat, HFD)–induced obese C57BL/6 mouse model was used toinvestigate the physiologic effect of DGAS-modified chestnut starch.

Results: Enzymatic modification of chestnut native starch by DGASincreased the proportion of RS, rendering it unavailable for hydrolysisby small-intestinal mucosal α-glucosidases. Structurally, the amyloseratio and branch chain of amylopectin in chestnut starch were increasedbyDGAS treatment. Furthermore, supplementation ofDGAS-modifiedchestnut starch to obese mice significantly reduced features of obesitycompared with HFD- or NCS-fed mice.

Conclusion: DGAS modification of chestnut starch increasesnondigestible RS, and this ameliorates diet-induced obesity via GPR43-mediated suppression of insulin signaling, thereby presumably reducingfat accumulation in white adipose tissue.

Funding SourcesThis research was supported by Basic Science Research Pro-

gram through the National Research Foundation of Korea (NRF)

funded by the Ministry of Science, ICT & Future Planning (NRF-2015R1C1A1A02036467).

Potato Starch Regulates Body Weight by Controlling GlucoseDelivery (P10-067)

Byung-Hoo Lee,1 Ho-Tak Jung,1 Jung-Min Seo,1 Eun-Sook Lee,2and Dong-Ho Seo2

1Gachon University, South Korea; and 2Korea Food ResearchInstitute

Objectives: The aim of this research was to investigate thephysiologic responses of various botanical sources of starch for differentglucose delivery properties by use of an in vitro mammalian digestivesystem and an in vivo animal model.

Methods:Corn, wheat, rice, and potato starcheswere used to test thedigestion properties ofmammalianmucosalα-glucosidase in an in vitroassay, and their physiologic effects were investigated in male C57BL/6mice.

Results: The results clearly demonstrated that potato starch causedthe lowest rate of glucose generation among the 4 different starches,and suggest that it can attenuate the postprandial glucose spike.Interestingly, a potato starch–based diet caused significantly (P < 0.05)less weight gain and fat accumulation in an animal model than dietsbased on the other starches, through increased insulin sensitivity.

Conclusion: Potato starch–based products can be applied to regu-late postprandial blood glucose concentrations, aiding in the control ofmetabolic diseases such as obesity and type 2 diabetes.

Funding SourcesThis research was supported by the Basic Science Research Pro-

gram through the National Research Foundation of Korea (NRF)funded by the Ministry of Science, ICT & Future Planning (NRF-2015R1C1A1A02036467).

Effects of White Potatoes and Other Carbohydrates Consumedwith Eggs on Short-Term Food Intake, Satiety, and GlycemicResponse in Children (P10-068)

Jennifer J Lee,1 Neil Brett,1 Jennifer Chang,1 Julia Totosy deZepetnek,2 and Nick Bellissimo1

1Ryerson University, Canada; and 2University of Regina, Canada

Objective: The aim of this study was to assess the effects ofconsuming commercially available white potato products within mixedmeals on short-term food intake (FI), subjective appetite, and glycemicresponse in children aged 9–14 y.

Methods: Using a within-subject repeated-measures design, 21children (age: 11.7 ± 0.4 y; body mass index percentile: 41.6 ± 2.2%)were exposed to 5 counterbalanced conditions, each separated by 7 d.After an overnight fast andwithin 2 h of awakening, childrenwere givenone of: isocaloric (450 kcal) treatment meals of eggs (246 kcal) servedwith fried French fries, mashed potatoes, or white beans; a cereal-basedtraditional breakfast (milk, cereal, and toast); or meal skipping. An adlibitum pizza meal was consumed 180 min following each condition,and parent-reported diet records were used to measure rest-of-dayFI. The total-day FI was calculated by summing the treatment meal,the pizza meal, and the rest-of-day FI. Subjective appetite via visual


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analog scales and capillary blood glucose were assessed at baseline,and throughout the 180 min between the condition and the pizzameal.

Results: The pizza FI (kcal) was lower (P< 0.001) after fried Frenchfries (1010 ± 73) and mashed potatoes (1039 ± 74) compared withtraditional breakfast (1257 ± 92) and meal skipping (1235 ± 74).The total-day FI (kcal) was lower (P = 0.03) after fried French fries(2228 ± 141) compared with traditional breakfast (2624 ± 137). Thesubjective average appetite was lower after mashed potatoes comparedwith traditional breakfast (P < 0.001). The blood glucose incrementalarea under the curve (mmol · min–1 · L–1) was higher after traditionalbreakfast (121.6 ± 22.9) compared with fried French fries (33.0 ± 22.6;P = 0.02) and meal skipping (–23.7 ± 12.5; P < 0.01).

Conclusions: Consumption of white potatoes with eggs suppressedlunch FI and total-day FI, subjective appetite, and glycemic responsewhen compared with a cereal-based traditional breakfast in childrenaged 9–14 y. Thus, consumption of white potato products at breakfastmay help children maintain a healthy body weight.

Funding SourcesThis study was supported by the Alliance for Potato Research and

Education (APRE).

Glucocorticoid-Transforming Growth Factor-β CrosstalkModu-lates Depot Differences in Adipogenesis: Potential Role in AdiposeExpansion and Remodeling (P10-069)

Mi-Jeong Lee, Richard Pickering, and Susan K Fried

Diabetes Obesity and Metabolism Institute, Icahn School ofMedicine at Mount Sinai, NY

Objectives:The aimof this studywas to understand themechanismsunderlying lower adipogenesis and fibrosis in visceral obesity byinvestigating the role played by glucocorticoid (GCs) modulation ofthe antiadipogenic transforming growth factor-β (TGFβ) signalingpathways in contributing to the depot differences in adipogenesis andfibrosis.

Methods: GC regulation of TGFβ signaling and adipogenesis wasassessed in primary human omental (Om) and abdominal subcuta-neous (Abdsc) adipose stem cells (ASCs). TGFβ activity and tissuefibrosis were determined in adipose tissue samples.

Results:OmASCs were less sensitive to the proadipogenic effects ofGCs and their ability to suppress TGFβ signaling activity. Basal SMAD2phosphorylation (SMAD2P) was elevated in Om compared with AbdscASCs and inversely correlated with differentiation degree of ASCs.Conditioned media (CM) from Om ASCs more potently inhibitedadipogenesis and stimulated SMAD2P than CM from Abdsc ASCs;culture of Om with dexamethasone downregulated the antiadipogeniceffects of CM and decreased TGFβ ligands in CM. Inhibition ofTGFβ signaling with SB431542 or silencing of both SMAD2 andSMAD3markedly improved adipogenesis. At the tissue level, SMAD2Pconcentrationswere higher inOm thanAbdsc, andwere associatedwithlower ASC adipogenesis and higher pericellular fibrosis.

Conclusion: Overproduction of TGFβ ligands and resistance toGCs conspire to limit the hyperplastic growth of visceral depots,and therefore their ability to expand in a healthy fashion. TargetingGC signaling to suppress TGFβ activity may provide therapeutic

opportunities to limit fibrosis and prevent metabolic dysfunction inobesity.

Funding SourcesNIH R01 DK080448, ADA 7-14-BS-059 to SKF, and funds from

Icahn School of Medicine at Mt. Sinai.

Vitamin D Regulates MED28 Expression and GlucoseMetabolism in Human Colorectal Cancer Cells (P10-070)

Ming-Fen Lee,1,2 Yu-Ting Weng,1,2 Nien-Tsu Hsieh,1 Chun-YinHuang,1 Chun-I Li,3 and Hsiao-Sheng Liu3

1China Medical University; 2Chang Jung Christian University,China; and 3National Cheng Kung University, China

Background: A growing body of literature suggests that vitamin Dmay limit cancer progression. MED28, a mediator subunit for tran-scriptional activation, was found to be highly expressed in colorectalcancer. Our laboratory previously reported that MED28 mediates cellmigration in human colorectal cancer cells, whereas vitamin D cansuppress this effect. Subsequently, we found that MED28 also regulatesglucose metabolism in human colorectal cancer cells.

Objectives: The objective of the current study was to investigatewhether vitamin D can suppress glucose metabolism by mediating theexpression of MED28 in colorectal cancer.

Methods: We employed a subcutaneous HT29 xenograftNOD/SCID mouse model and 2 human colorectal cancer cell lines,HT29 and SW480, to study the effect of vitamin D on glucosemetabolism. We treated HT29 and SW480 with 0–100 nM ofcacitriol (1,25-dihydroxyvitamin D3), evaluated the uptake of glucose,and analyzed the expression of MED28 and glycolysis-associatedenzymes and transporters by reverse transcriptase-polymerase chainreaction and Western blotting. The mice were administered calcitriol(0.5–1 µg) or vehicle only by intraperitoneal injection every other day.The animal study continued for 30 d, and the animal use protocol wasapproved by the Institutional Animal Care and Use Committee. Thesubcutaneous tumors were analyzed for the expression of MED28 andglycolysis-associated enzymes and transporters.

Results: Calcitriol reduced the expression of MED28, along withthe gene products involved in glucose metabolism, including glucosetransporters and glycolytic enzymes, in HT29 and SW480 cells.Regarding the xenograft model, the volume of the tumors was smallerin the calcitriol group than in the control group. The expression levels ofMED28, glucose transporter Glut1, and a few glycolytic enzymes werelower in the calcitriol group in the xenografts.

Conclusions: Calcitriol may attenuate the MED28-mediated War-burg effect in human colorectal cancer cells.

Funding SourcesThis work was supported in part by the grants MOST105-

2320-B-309-001 and MOST106-2320-B-039-062-MY3 to M-FL, andMOST105-2815-C-309-006-B to Y-TW.

Obesogenic Environment Factors Related to the Incidence ofObesity in Korean Children (P10-071)

Myoung Sook Lee, SongJoo Kang, Soyoung Sung, and Minji Lee

Sungshin Women’s University, South Korea


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Background: Obesity is an imbalance between energy intake andexpenditure, and is a risk factor for various metabolic diseases. Beingobese starts with an increase the amount of body fat and leadsto adipocyte hypertrophy. Obesogenic environments (OEs) are allenvironmental factors which induce people to become obese.

Objective: Our objective was to find out which OEs, includingnutrients, have an effect on weight gain in Korean children.

Methods: For our study, 7 elementary schools in Seoul wererandomly selected and 770 Korean children in those schools (600normal weight, 170 obese), aged 8–9 y, participated in the study.Participants were surveyed over a range of parameters, including bodymass index (BMI) and physical activity. Blood triglyceride (TG), HDLcholesterol, and LDL cholesterol concentrations were measured. Forquantitative analysis of nutrients, dietary intakes (24 h for 3 d) wereanalyzed by CanPro4.0 software. All statistical tests were 2-tailed, andP-value

Results: We confirmed the differences between normal and obesechildren in 2007–2010 and 2009–2012 cohort studies. In the 2007–2010 cohort study, the BMI was increased and the HDL cholesterolconcentration decreased in obese children compared with those ofnormal weight. TG concentrations were increased in obese boys whohad a high carbohydrate intake compared with girls. In normal-weightchildren, HDL cholesterol and TG concentration were correlated withphysical activity. In the 2009–2012 cohort study, obese biomarkerssuch as BMI, LDL cholesterol, and HDL cholesterol were significantlyincreased and the intakes of minerals and vitamins decreased.

Conclusion:OE factors influenced the obesity incidence in 2 sets ofcohort studies. We believe that the findings of this study contribute tothe prevention of the early onset of chronic diseases caused by obesityin children.

Funding SourcesFunded by a grant from the Korea Health Industry Development

Institute of the Ministry of Health & Welfare, Republic of Korea(#HI17C0863).

Plasma Proteins Associated with Lipoproteins Reflect Rolesbeyond Lipid Metabolism in Nepalese School-Aged Children (P10-072)

Sun Eun Lee,1 Kerry Schulze,1 Christine P Stewart,2 Robert NCole,3 Lee S-F Wu,1 Abdulkerim Eroglu,1 James D Yager,1 JohnGroopman,1 Parul Christian,1 and Keith P West1

1Johns Hopkins Bloomberg School of Public Health, MD;2University of California, Davis; and 3Johns Hopkins School ofMedicine, MD

Background: Circulating lipoproteins play central roles in lipidtransport, energy metabolism, and cardiovascular function. However,little is known about the diversity of metabolic associations oflipoproteins in childhood that may affect cardiovascular health later inlife, especially in impoverished societies.

Objectives: The aim of this study was to apply quantitativeproteomics to detect plasma proteins associated with lipid profiles inschool-aged children in rural Nepal.

Methods: The relative abundance of 982 plasma proteins wasquantified by tandem mass spectrometry in a cohort of 500 6- to 8-y-old children. Associations with HDL, LDL, and triglyceride (TG)

concentrations were assessed by linear mixed-effects models, applyinga false discovery rate, q < 0.05.

Results: The prevalence of high LDL (≥130 mg/dL), high TG(≥100 mg/dL), and low HDL (<40 mg/dL) concentration was 1%,41%, and 90%, respectively. Among 53 proteins associated with HDLconcentrations, positive correlates (n = 35) included apolipoprotein(APO) A1 (r= 0.79, q= 1.19× 10–92) and 8 other apolipoproteins (A2,A4,C1, C3,D,M, F, and serumamyloidA4) involved in the integrity andmetabolism of HDL; enzymes required for HDL remodeling, choles-terol efflux, and inhibition of LDL oxidation (phospholipid transfer pro-tein, lecithin-cholesterol acyltransferase, and paraoxonase 1); plasmacarriers of vitamins A and E (retinol binding protein 4, transthyretin,and afamin); and proteins regulating cell adhesion, differentiation,and growth. A negative HDL proteome (n = 18) mainly includedproteins of inflammatory and innate immune responses. Among 12LDL-associated proteins, APO B, the primary component of non-HDLcholesterol, was most strongly correlated with LDL (r = 0.70; q = 6.45× 10–47). APO Cs (C2, C3, and C4) and E, and cathelicidin were amongpositive correlates of TG (r= 0.50–0.53; q= 1.56× 10–13–8.92× 10–17).

Conclusions: Plasma proteomics analyses revealed a highly diver-sified HDL lipoproteome with anti-inflammatory, antioxidative, fat-soluble vitamin transport, angiogenic, and tissue remodeling properties.Childhood plasma lipoproteomes include those with both classical andnovel roles which may have prognostic value for cardiovascular diseaserisk later in life.

Funding SourcesBill & Melinda Gates Foundation (OPP5241 and GH614).

Mediation Effects of Biochemical Markers on Relationship be-tween Usual Dietary Intakes and Risk of Metabolically UnhealthyObesity in Chinese Adults (P10-073)

Kaifeng Li,1 Yongkang Luo,2 Qi’an Han,2 and Beiwei Zhu3

1The Beijing Advanced Innovation Center for Food Nutrition andHuman Health, China; 2China Agricultural University; and 3DalianPolytechnic University, China

Objective:The aim of this study was to explore themediation effectsof metabolic markers on the relation between usual food intakes andrisk of metabolically unhealthy obesity in Chinese adults.

Methods:Dietary and demographic data from theChinaHealth andNutrition Survey in 2000 were used as a baseline (n = 8447). K-meanscluster analysis was used to identify metabolic healthy and unhealthysubjects in 2009 using data of biochemistry biomarkers (glucose, HDL,triglycerides, and high-sensitivity C-reactive protein). Bayesian analysiswas used to estimate ORs and 200 bootstrap usual intake samples wereused to obtain SEs and 95%CIs of indirect, direct, and total effects usingMplus.

Results: A total of 210 participants were identified as metabolicallyunhealthy obese in 2009, with an incident rate of 4.8%. A total energyincrease in red meat and organ meat intakes of 1% increased the riskof metabolically unhealthy obesity significantly, by 3% (OR: 1.03; 95%CI: 1.02, 1.04), through the mediation effects of triglycerides, withno significant total or direct effects being observed. A 1% increasein total energy from egg and egg product intakes increased the riskof metabolically unhealthy obesity by 11% (OR: 1.11; 95% CI: 1.01,


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1.28) through the total effects of HDL, and the effects became non-significant after further adjustment for body mass index, total income,urbanization index, blood pressure, smoking habits, and physicalactivity.

Conclusion: Intakes of red meat and organ meat exert an indirecteffect on risk of metabolically unhealthy obesity through circulatingtriglyceride concentrations. Our study highlights the importanceand necessity of carrying out mediation analysis in a nutritionstudy.

Funding SourcesThis research uses data from the China Health and Nutrition Survey

(CHNS). This study is supported by the Beijing Advanced InnovationCenter for Food Nutrition and Human Health (201,502,910,110,942)and the National Nature Science Foundation of China (31,401,520).

Associations of Plasma Free Amino Acids with Hyperuricemia ina 6-y Cohort Study of Chinese Men andWomen (P10-074)

Xu Lin,1 Feijie Wang,1 Liang Sun,1 and Rong Zeng2

1Institute of Nutritional Sciences and 2Institute of Biochemistry andCell Biology, Chinese Academy of Sciences

Background:Hyperuricemia is known as one of the risk factors forgout and cardiometabolic diseases. Certain amino acids play importantrole(s) in biosynthesis of purine and subsequent formation of uric acids.Different relations between amino acid profiles and hyperuricemiaor gout were reported by previous cross-sectional studies; however,evidence from prospective studies is limited.

Objective:This study therefore aimed to investigatewhether specificamino acid profiles are associatedwith the risk of hyperuricemia, as wellas potential modifying factors, in a 6-y Chinese cohort.

Methods: The current study included 1672 community-living Chi-nese men and women, aged 50–70 y, without baseline hyperuricemia.Baseline plasma amino acids were measured by GC-MS. Incidenthyperuricemia was defined as the occurrence of plasma uric acid ≥7.0mg/dL for men or ≥6.0 mg/dL for women by the end of the 6-y follow-up. Logistic regression was applied to assess the associations of plasmaamino acids with incident hyperuricemia, after adjustment for baselinerisk factors. A false discovery rate method was used to control formultiple testing.

Results: The 6-y incidence of hyperuricemia was 24.6%. Elevatedcysteine and threonine were showed to be positively associated withincreased incident hyperuricemia (RR: 1.25; 95% CI: 1.15, 1.36;P < 0.001; and RR: 1.13; 95% CI: 1.04, 1.22; P = 004 per 1 SDincrement of cysteine and threonine, respectively) in both men andwomen, after multivariable adjustment. Notably, positive associationsbetween phenylalanine and glutamine concentrations and incident hy-peruricemia were only detected in men, whereas an inverse associationbetween serine concentrations and incident hyperuricemia was onlyevidenced in women (all P-interaction < 0.05).

Conclusions: In this prospective cohort study, we found thatincreased concentrations of cysteine and threonine, mainly involvingincreased oxidative stress, were associated with higher incidence

of hyperuricemia in both men and women. However, sex-specificassociations were observed in the case of phenylalanine and glutaminefor men and serine, the precursor of uric acid, for women. It remainsto be elucidated whether the specific amino acids could explain, at leastpartially, the sexual differences in uric acid metabolism.

Funding SourcesThis work was supported by theMinistry of Science and Technology

of China (2016YFC1304903 and 2017YFC0909700); the NationalNatural Science Foundation of China (81,471,013 and 81,700,700); andthe Chinese Academy of Sciences (ZDBS-SSW-DQC-02).

High-Fat Diets Containing Different DHA-to-EPA Ratio AlterFatty Acid Composition and Attenuate Hepatic Lipid Accumulationin ApoE–/– Mice (P10-075)

Liang Liu,1 Zhiyong Gong,1 and Guoxun Chen2

1Wuhan Polytechnic University, China; and 2University of Ten-nessee at Knoxville

Objective: We have reported that dietary various ratios of docosa-hexaenoic acid (DHA) to eicosapentaenoic acid (EPA) have protectiveeffects on high-fat diet (HFD)–induced liver damage in mice. The aimof this study was to determine if the protective roles are associated withthe alteration of hepatic fatty acid composition in mice.

Methods:Using HFDt-fed ApoE–/– mice as the animal model, dietsof various ratios of DHA:EPA (2:1, 1:1, and 1:2) with an ω-6:ω-3(n–3:n–6) ratio of 4:1 were prepared using fish and algae oils enrichedin DHA and/or EPA, and sunflower seed oils for comparison with theHFD.

Results:DHA:EPA supplementation significantly decreased hepaticlipid deposition, serum lipid profile, inflammatory reactions, lipidperoxidation, and expression of adipogenesis-related proteins andinflammatory factors as compared with the HFD group. In the variousratios of the DHA:EPA groups, we found increases in total saturatedfatty acids (SFAs), total ω-6 series polyunsaturated fatty acid (PUFAs)and total ω-3 series PUFAs in the liver compared with the HFDgroup. Also, the amount of monounsaturated fatty acids (MUFAs)and the ratio of ω-6:ω-3 showed remarkably decreases after DHA:EPAsupplementation. However, the 2:1 DHA:EPA group showed lowerserum triglycerides (TG), total cholesterol (TC), LDL cholesterol,interleukin-1β , aspartate aminotransferase, alanine aminotransferase,and alkaline phosphatase concentrations, and there were higher serumadiponectin concentrations with all 3 DHA:EPA ratios. No significantdifferenceswere found in the expression of hepatic inflammatory factorsor hepatic lipid metabolic-related genes in the 3 DHA:EPA groups.There were also no significant differences between the 3DHA:EPA ratiogroups for SFAs, MUFAs, PUFAs, and ω-6:ω-3 ratio.

Conclusions: Our data demonstrate that dietary DHA and EPAalters the composition of hepatic fatty acids and attenuates HFD-induced hepatic lipid accumulation, although no significant differenceswere found in the composition of hepatic fatty acids or the expressionof inflammatory factors or hepatic lipid metabolic-related genes in the3 DHA:EPA groups.


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Abdominal Obesity and Risk of Developing Rheumatoid Arthri-tis in Women (P10-076)

Bing Lu,1,2 Jeffrey Sparks,1,2 Sara K Tedeschi,1,2 Susan Malspeis,1Karen H Costenbader,1,2 and Elizabeth Karlson1,2

1Brigham and Women’s Hospital, MA; and 2Harvard MedicalSchool, MA

Objective: Being overweight or obese increases the risk of rheuma-toid arthritis (RA) among women. The aim of this study was toinvestigate whether abdominal obesity predicts RA risk in 2 largeprospective cohorts, the Nurses’ Health Study (NHS) and Nurses’Health Study II (NHSII).

Methods: We followed 48,919 women in NHS (aged 40–67 y in1986) and 47,220 women in NHSII (aged 29–48 y in 1993) without RAat baseline. Lifestyle and environmental exposures and anthropometricmeasures were collected through biennial questionnaires. IncidentRA cases were identified using the previously validated connectivetissue disease screening questionnaire, followed by a medical recordreview. RA serologic status was determined by positive rheumatoidfactor or anticitrullinated peptide antibodies in the medical record.Abdominal obesity was measured using waist circumference (WC)

TABLE 1

reported in 1986, 1996, and 2000 in NHS and 1993 and 2005 in NHSII.The cutoff point for abdominal obesity (WC ≥88 cm) was based onWHO recommendations. Using pooled data from the 2 cohorts, weestimatedHRs for RA risk using time-varyingCox proportional hazardsmodels. We repeated analyses restricted to young and middle-agedwomen (age ≤55 y).

Results: During 28 y of follow-up, we identified 807 incident RAcases (510 in NHS, 297 in NHSII). Age-adjusted incidence rates were33 per 100,000 person-years in NHS and 28 per 100,000 person-yearsin NHSII. Women with WC >88 cm had increased RA risk comparedwith women with WC

Conclusions: In this prospective cohort study of women followedup to 28 y, abdominal obesity was significantly associatedwith increasedrisk of developing RA. Abdominal obesity conferred the greatest risk forseropositive RA among women ≤55 y old independent of body massindex.

Funding SourcesThis study was supported by the NIH through grants AR061362,

AR049880, AR052403, AR047782, AR059073, AR066953, AR069688,AR066109, and CA186107, CA176726, CA49449, CA67262.


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Dyslipidemia Risk and Saturated Fat Intake from Records ofPatients who Receive Precision Ketogenic Therapy (P10-077)

Jurate Lukosaityte and Peggy Borum


Background: Precision ketogenic therapy (PKT) is a high-fat, low-carbohydrate, adequate-protein diet that is used to treat seizures inpatients with refractory epilepsy. Research has suggested that ketogenicdiets may increase the risk for dyslipidemia.

Objectives: The objective of this work was to determine if saturatedfat intake was higher among patients who developed dyslipidemia onPKT.

Methods: This was a cross-sectional analysis of patients who werereferred to the University of Florida PKT Program from 1995 to 2016.Caregivers were provided with recipes and were instructed to weigh allfood to the nearest one-tenth of a gram. Patients were included for thisanalysis if they had a fasting lipid panel and diet records at any timepoint on PKT through 24 mo. The Mann-Whitney U test was used tocompare each lipid parameter from baseline to 6, 12, 18, and 24 mo onPKT. The PKT ratio (grams of fat to grams of protein and carbohydrate)and saturated fat (SFA) intake (percentage of total fat) were comparedbetween records of patients onPKTwhohad acceptable or unacceptablelipids.

TABLE 1 SFA intake (percentage of total fat) over time on PKT forpatients with acceptable or high TC

Results: Patients remained on PKT for a median of 1.2 y (n = 55).There were no significant differences in total cholesterol (TC), non-HDL, or LDL frombaseline to any time point on PKT.HDLwas lower at12 and 18mo on PKT compared with baseline (P< 0.01), but remainedacceptable for the majority of patients. Triglycerides (TG) were higherat 6, 12, and 18 mo on PKT compared with baseline for patients whowere <10 y old (P < 0.01). Diet intake data were available from 13,580records collected on PKT. There were no differences in PKT ratio intakefor patients with acceptable or unacceptable lipids. SFA intakewas lowerfor patients with acceptable TC at 6 and 12 mo on PKT compared withpatients with high TC (Table 1). There were no differences in SFA intakebetween patients with acceptable or high TG.

Conclusions: Lowering SFA intake can improve TC concentrationsfor patients who receive PKT. Future work should explore the relationbetween dietary fatty acid intake and the increase in TG patients onPKT.

Funding SourcesUniversity of Florida Foundation (KetoGator, Fund ID:

01,8555). The research reported in this publication was supportedby the University of Florida Clinical and Translational ScienceInstitute, which is supported in part by the NIH National Centerfor Advancing Translational Sciences under award numberUL1TR001427.


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Effects of Feeding Waste Water Algae on Mouse Metabolome(P10-078)

Yiwei Ma, Chi Chen, Roger Ruan, Wenguang Zhou, Paul Chen,Pedro Urriola, Gerald Shurson, and Qingqing Mao

University of Minnesota, Twin Cities

Background: Wastewater from animal and food production com-monly contains significant amounts of nutrients. Without properprocessing, nutrient-rich wastewater is an environmental hazard.Cultivating algae in wastewater reduces pollution and also provides aneconomic source of feed ingredients.

Objective: The objective of this study was to examine in detail themetabolic events occurring after feeding wastewater algae to animals.

Methods: In this study, 5 groups of young male mice (n = 8/group)were fed chows containing 0%, 5%, and 10% green algae (Chlorellavulgaris) grown in wastewater from a slaughterhouse and from a dairyprocessing facility for 28 d. Both sources of algae have elevated proteincontents that are rich in glycine and alanine. The metabolic effects ofthe wastewater algae were investigated by growth performance, bloodchemistry, and LC-MS-based metabolomics.

Results: The results showed that daily weight gain, food intake,and blood chemistry, including glucose, triacylglycerol, cholesterol, andblood urea nitrogen, were not affected by algae feeding. Metabolomicanalysis of urine, serum, and feces samples identified both dose-dependent and source-specific metabolic changes. In urine, the con-centrations of B vitamins were significantly increased by both algae.In serum, glycine, alanine, and 2 essential amino acids, phenylalanineand glycine, were increased by both algae. Short-chain fatty acids infeces were only increased by the 10% slaughterhouse algae. Muricholicacid and deoxycholic acid in feces were decreased by both the 10%slaughterhouse and the 10% dairy algae, whereas lithocholic acid wasdecreased only by the 10% slaughterhouse algae.

Conclusions: Overall, feeding on wastewater algae did not elicitany apparent negative effect on young mice. The observation of algae-elicited changes in amino acid and microbial metabolism could serveas a foundation for further mechanistic investigations on the biologicaleffects of feeding on wastewater algae.

Total Dietary Protein, Not Plant versus Animal Amino AcidComposition, Determines Health Benefits (P10-079)

Michael R MacArthur, Jose Trevino-Villarreal, Justin Reynolds,Kaspar M Trocha, Sarah J Mitchell, Katia Chadaideh, YohannGrondin, Rachel Carmody, and James Mitchell

Harvard School of Public Health, MA

Objective: Studies show divergent associations of plant vs. animalprotein on disease risk factors, with plant protein having a protectiveeffect. The objective of this study was to investigate whether thesedifferential associations can be explained by different amino acid (AA)compositions of plant- vs. animal-based diets.

Methods: Food items were analyzed using the USDA NutrientDatabase SR 28 (NDB). To analyze AA composition of dietary recordsfrom NHANES, data from SR28 were linked to NHANES FNDDS fooditems using SR codes. Mouse experiments were performed with 16-wk-old male B6D2/F1 mice. Semipurified diets were made with protein-free powder based on Research Diets D12450B base and differentamounts of crystalline AA according to plant or animal AA ratiosdetermined by USDANDB analysis. Naturally sourced diets were madeby homogenizing plant- or animal-derived whole food items with 1%agar in a blender to form a solid diet.

Results: Per gram, all AAs were higher with large effect size inanimal vs. plant foods, but as a ratio of total AAs, only 4 weresignificantly different with large effect size (P < 0.05; Cohen’s d: >0.8).Individualswho ate animal products consumedmore protein than thosewho did not (0.77 vs. 0.61 g/kg, P < 0.001), but with no differences indietary AA ratios. The protein intake decile was positively associatedwith blood glucose, insulin, and waist circumference in multivariatelinear models adjusted for confounders (P < 0.05). To test if proteinquantity or AA composition drives health benefits, semipurified dietswere tested in mice with AA ratios modeled after plant vs. animalfoods from theNDB. There were significantmain effects of total proteinacross 6 levels, but not AA composition, on FGF21, adiponectin, fastingglucose, and body weight (P < 0.05). When fed 5, 10, or 18% proteindiets made with plant- or animal-based whole-food items, there weresignificant main effects of total protein, but not protein source on thesame parameters.

Conclusion:Animal- vs. plant-based food sources and diet patternshave similar AA profiles but differ in total protein content. Lowerprotein diets are associated with improved chronic disease risk factorsin humans and in controlled mouse studies independent of plant vs.animal AA composition or source. Reduced dietary protein content islikely a contributor to the benefits of plant-based diets.

Funding SourcesFunding for this work was provided by the Charoen Pokphand

Corporation.


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FIGURE 1 A) Animal-based food items contain more protein per gram weight, but only lysine, methionine, histidine and serine aresignificantly different when assesed as a proportion of total amino acids. B) When asssessed as a proportion of total amino acids, there areno significant differences with large effect size (Cohen’s d > 0.08) between the 1st and 10th decile of total protein intake. C) Protein intakedecile postively correlates with fasting blood glucose. D) In a controlled mouse study, fasting glucose is increased with greater proteinintake, but is not affected by plant vs animal amino acid composition. E) FGF21 is suppressed by increasing protein intake of mice,independent of whether naturally sourced protein is from animal or plant-based products.

Resistant Starch Type 4 Incorporated into a Breakfast Bar De-creasesPostprandialGlucose and InsulinResponses:ARandomized,Controlled, Crossover Study (P10-080)

Eunice Mah,1 Vicenta Garcia-Campayo,2 and DeAnn J Liska1

1Biofortis Inc.; and 2Cargill, Inc.

Background:Resistant starch type 4 (RS4) are resistant starches thathave been chemically modified. Similar to other forms of dietary fiber,RS4 are not digested by human enzymes.

Objective: The aim of this study was to test the hypothesize thatreplacement of fully digestible carbohydrates with RS4 in a bakedbreakfast bar will decrease postprandial glycemic and insulinemicresponses in healthy adults.

Methods: In this double-blind, randomized, placebo-controlledcrossover study, 21 healthy adults (10 males, 11 females; age: 20–45y; body mass index: 19.3–27.0 kg/m2) consumed a baked breakfastbar containing 28 g of the RS4 (Actistar75,330 from Cargill, Inc.) ora control bar that was matched for fat, protein, and total carbohydratecontent, delivering 32 g and 4 g of dietary fiber, respectively (verifiedby the AOAC 991.43 method for dietary fiber). Capillary blood glucoseand intravenous blood insulin were measured at t = –15, –5, 15, 30,45, 60, 90, and 120 min, where t = 0 min was the start of the studyproduct consumption. The primary outcome was the incremental areaunder the curve (iAUC0–120 min) for capillary glucose, calculated usingthe trapezoidalmethod.Other outcomes included insulin iAUC0–120 min,

glucose and insulin maximum concentration (Cmax), and time to Cmax

(Tmax). Repeated measures ANCOVA was used to assess differencesbetween active and control foods using SAS for Windows (version 9.4).

Results: Median glucose iAUC0–120 min was 22% lower (P < 0.05)and median insulin iAUC0–120 min was 37% lower (P < 0.05) followingconsumption of the RS4 food compared with the control food. Therewere no statistically significant differences (P> 0.05) between foods forglucose and insulin Cmax and Tmax.

Conclusion: RS4 incorporated into a baked breakfast bar signifi-cantly decreased postprandial iAUC for glucose and insulin responsesin healthy adults. The results suggest that replacement of digestiblecarbohydrate with RS4 is a promising strategy for postprandial bloodglucose management.

Funding SourcesCargill, Inc.

Influence of Age, BMI, and Sex on Breath Hydrogen Productionfollowing Consumption of a Fermentable Fiber: A Pilot Study (P10-081)

Eunice Mah, Jose Rotor, and DeAnn J Liska

Biofortis Inc.

Objective: The hydrogen breath test is a simple and noninvasivemethod to estimate the available energy value of fibers. Breath hydrogenproduction is dependent on the composition of the gastrointestinalmicrobiota, which is known to be influenced by age, body mass index


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(BMI), and sex. However, the influence of these factors on breathhydrogen production following fiber consumption is unknown. Theaim of this study was to determine that influence.

Methods: In this open-label, single-arm, pilot study, 40 healthyadults (19 males, 21 females) consumed a single serving of 20 g offructo-oligosaccharide (FOS; NutraFlora FOS Pure Powder fromNOWFoods) dissolved in 250 mL of water. Breath hydrogen concentrationwas measured using a BreathTrackerTM Analyzer SC (QuinTronInstrument Company, Inc.) at t = –0.25, 1, 2, 3, 4, 5, 6, 7, 8, 9, and10 h, where t = 0 min was the start of FOS consumption. Subjectswere instructed to avoid vigorous physical activity, alcohol, and foodsthat cause gastrointestinal distress 24 h prior to testing, and consumedstandardized low-fiber meals the evening prior to and during the dayof testing. The primary outcome was the positive incremental areaunder the curve (iAUC-0.25–10 h) for breath hydrogen, calculated usingthe trapezoidal method. Comparisons between age groups (18–34 vs.35–49 vs. 50–65 y, n = 10/group), BMI categories (18–34 y; BMI: 18.5–29.9 vs. ≥30 kg/m2; n = 10/category), and sex were done using SAS forWindows (version 9.4).

Results: Breath hydrogen iAUC-0.25–10 h tended (P = 0.0693) to begreater in female subjects than in male subjects following consumptionof FOS. There were no significant differences between age groups(P = 0.7308) and BMI categories (P = 0.8756) for breath hydrogeniAUC-0.25–10 h. There was an 8-fold difference for iAUC-0.25–10 h betweenthe lowest and highest responders, indicating a large intrasubjectvariation in breath hydrogen production.

Conclusion: Breath hydrogen production following the consump-tion of 20 g of FOS tended to be higher in women than in men,and was not significantly affected by age or BMI. Additionally, breathhydrogen production was highly variable among subjects. Theseresults suggest that sex and intraindividual variation are importantconsiderations when designing studies measuring breath hydrogenproduction following consumption of indigestible carbohydrates.

Funding SourcesBiofortis, Inc.

CornOil Consumption Improves the Plasma Lipid Profile versusCoconut Oil in Men and Women with Above-Desirable Levels ofCholesterol (P10-082)

Kevin C Maki,1 Wendy Hasse,2 Mary Dicklin,1 Marjorie Bell,1Mary Buggia,3 Martha Cassens,4 and Fulya Eren4

1Midwest Biomedical Research/Center for Metabolic and Car-diovascular Health, MO; 2Great Lakes Clinical Trials; 3MB ClinicalResearch; and 4ACH Food Companies Inc.

Objective:The aimof this trial was to assess the effects of consumingfoods made with corn oil vs. coconut oil on LDL cholesterol andother aspects of the fasting lipoprotein lipid profile, and a marker ofinflammation in men and women with above-desirable concentrationsof LDL cholesterol.

Methods: In this randomized, crossover study, subjects with afasting LDL cholesterol concentration of 115–<190 mg/dL and triglyc-erides (TG) ≤375 mg/dL were randomly assigned to consume foodsproviding 4 tablespoons/d of corn oil or coconut oil (muffins and rolls)for 4wk. Subjects received diet instruction on the incorporation of study

foods to maintain their habitual energy intake. Fasting lipids and high-sensitivity C-reactive protein (hs-CRP) were assessed at baseline andafter each diet condition. Differences in responses between conditionswere evaluated using repeated-measures ANCOVA with the baselinevalue as a covariate.

Results: Subjects (n = 23; 10 men and 13 women) had a meanage of 45.0 y and mean body mass index of 27.8 kg/m2. Medianbaseline concentrations of LDL cholesterol, non-HDL cholesterol, totalcholesterol (TC), HDL cholesterol, and TG were 123, 144, 188, 46.0,and 92.5 mg/dL, respectively, and the mean TC to HDL cholesterolratio was 4.2. Changes from baseline for corn oil and coconut oilconditions, respectively, were: LDL cholesterol (–2.7% vs.+4.6%), non-HDL cholesterol (–3.0% vs. +5.8%), TC (–0.5% vs. +7.1%), HDLcholesterol (+5.4% vs. +6.5%), TG (–2.1% vs. +6.0%), and TC:HDLcholesterol (–4.3% vs. –3.3%). Non-HDL cholesterol responses weresignificantly different between corn and coconut oils (P = 0.034);differences in LDL cholesterol and TC between conditions approachedsignificance (P = 0.062 and P = 0.057, respectively). Median hs-CRPincreased by 3.8% from a baseline value of 1.6 mg/L with coconut oil vs.0.0% with corn oil (P = 0.338).

Conclusions:These results indicate that, when incorporated into thehabitual diet, consumption of foods providing 4 tablespoons/d of cornoil, which contains a higher quantity of plant sterols than other cookingoils and is rich in polyunsaturated fatty acids, produced amore desirableplasma lipid profile and reduced non-HDL cholesterol compared withcoconut oil (high in 8–12 carbon saturated fatty acids) in men andwomen with above-desirable concentrations of circulating cholesterol,and did not elevate hs-CRP concentrations.

Funding SourcesThis study was funded by ACH Food Companies, Inc. (Oakbrook

Terrace, IL).

Acute Mild Cold Exposure Induces a Measurable Increase inEnergy Expenditure in Normal Weight and Overweight or ObeseIndividuals through Nonshivering Thermogenesis (P10-083)

Alexandra Martin, Soonkyu Chung, Brian Smith, and KarstenKoehler


Background: When ambient temperature decreases beyond thethermoneutral zone, heat generation and a concomitant increase inenergy expenditure (EE) are required to maintain core temperature.The increase in EE occurs in skeletal muscle, where shivering leads tounsustainable increases in EE, and brown adipose tissue (BAT), wherenon-shivering thermogenesis (NST) increases EE through prolongedheat production. However, measuring these organs’ separate metabolicactivities in vivo is challenging.

Objectives: The goal of the present study was to quantify theadaptive increase in whole-body EE as a result of NST in response tomild cold exposure (21ºC) in normalweight and overweight or obese in-dividuals. We hypothesized that EE increases upon mild cold exposurewithout shivering, suggesting a quantifiable contribution of BAT to EE.

Methods: Whole-body resting EE (REE) was measured at roomtemperature (24ºC) and mild cold exposure (21ºC) in normal weight(NW; n = 10) and healthy overweight or obese (OW; n = 6)individuals. Measurements were conducted via indirect calorimetry


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following a 12-h fast during 2 separate visits in random order withdiet replication before each visit. Body composition and supraclavicularheat production (SHP), assessed via infrared thermography, were alsomeasured.

Results: NW and OW participants did not differ in age (P = 0.31).Per study design, body weight (P < 0.001), fat mass (FM; P < 0.001),and fat free mass (FFM; P = 0.037) were significantly greater amongOW than among NW. Whole-body REE at 24°C, normalized for FFM,was similar between NW and OW (26.0 ± 0.3 vs. 25.6 ± 0.7 kcal/kgFFM; P = 0.86). At 21°C, REE increased in both NW (+5.7% ± 0.5%,P = 0.002) and OW (+10.7% ± 2.0%, P = 0.037), but the increase inREE was not significantly different between OW and NW (P = 0.37).When comparedwith 24°C, SHP at 21°C increased to a similar extent inboth NW (+1.2± 0.2°C, P= 0.03) and OW (+1.4± 0.1°C; P< 0.001).

Conclusions:NST is inducible uponmild cold exposure to a similarextent in both NW and healthy OW individuals, and occurs with aconcomitant increase in SHP, possibly reflecting BAT activity. Thissuggests that mild cold exposure may be a practical method to quantifyacute BAT activity in terms of kilocalorie expenditure.

Funding SourcesUSDA National Institute of Food and Agriculture.

Effect of High- or Standard-Protein Low-Calorie Diets on Glu-cose Metabolism and Adipokines in Diabetic Subjects Who AreOverweight or Obese (P10-084)

Rocío Mateo-Gallego,1,2,3,4 Victoria Marco-Benedí,1,2,3 SofiaPérez-Calahorra,1,2,3 Ana M Bea,1,2,3 Itziar Lamiquiz-Moneo,1,2,3Lucia Baila-Rueda,1,2,3 Ana Cenarro,1,2,3 and FernandoCiveira1,2,3,4

1Hospital Universitario Miguel Servet, Spain; 2Instituto de Investi-gación Sanitaria Aragón (IIS Aragón), Spain; 3CIBERCV, Spain; and4Universidad de Zaragoza, Spain

Objective: The aim of this study was to assess the effect of low-calorie diets with normal (18%) vs. high (35%) protein (mainly comingfrom a lean animal source) composition on glucose metabolism andadipokine concentration in overweight and obese subjects with type 2diabetes.

Methods:A total of 73 subjects (43.8%men; age: 55.6± 8.37 y; bodymass index: 32.8 ± 3.67 kg/m2) were randomized to follow for 6 mo 1of 2 calorie-restricted diets with the following distributions of caloriesfrom protein, carbohydrates, and fat: 1) 18%, 52%, and 30%; and 2)35%, 35%, and 30%. Anthropometric, clinical, biochemical (includingadipokine concentration), dietary, and physical activity assessment wasperformed at baseline and after 3 and 6 mo of intervention.

Results: Weight loss decreased homogeneously among the di-ets. Glycemic control showed a greater improvement regardless ofweight loss in subjects consuming the 35% protein diet, includingglucose, insulin, homeostaticmodel assessment insulin resistance index(HOMA-IR), and glycated hemoglobin (HbA1c). HOMA-IR decreasedby 23.4%± 27.7% and 37.7%± 21.3% after 3 mo (P= 0.023 comparingboth diets), and by 20.6% ± 34.6% and 41.7% ± 18.1% after 6 mo(P = 0.004) with the 18% and 35% protein diets, respectively. Retinolbinding protein 4 and leptin significantly decreased in both diets,although no differences were found between the diets. Adiponectinsignificantly increased in both diets; it changed by 6.05% and 29.9% at

3 mo in the 18% and 35% protein diets, respectively (P = 0.167), andby 23.7% and 53.5%, respectively, at 6 mo (P = 0.219). Adiponectinvariation was inversely correlated to HbA1c (R = –0.431, P = 0.043)and HOMA-IR (R = –0.262, P = 0.047) changes at 6 mo.

Conclusions: A restricted diet with 35% protein energy led to agreater improvement in glucose metabolism, especially as observedin HOMA-IR, irrespective of weight loss. Adiponectin concentrationsshowed a higher increase after consumption of the 35% protein diet, al-though statistical significance was not found. This change was inverselyassociated to HbA1c, HOMA-IR, and insulin variations. Our findingsdemonstrate that high-protein diets improve glycemic parametersregardless of weight loss in diabetic subjects. However, this effect seemsnot to be explained by changes in plasma concentrations of adipokines.

Funding SourcesThis study was supported in part by 3 grants from the Carlos III

Research Institute: CIBERCV (cosupported by the European RegionalDevelopment Fund (ERDF) which is allocated by the European Union;IIS16/0114), PI13/0,2507, and PI15/0,1983.

Leukocyte Gene Expression within a Postnatal Rat Undernutri-tion Model (P10-085)

Aaron Mehus and Matthew Picklo

USDA-ARS

Background: Childhood undernutrition is a global issue. Postnatalnutritional status impacts immune system development, and emergingevidence suggests important roles of dietary n–3 (ω-3) polyunsaturatedfatty acids (PUFAs) in immunomodulation. Knowledge is lacking asto which genes are sensitive to dietary n–3 PUFAs in leukocytes fromyoung animals.

Objectives: The primary objective of this study was to determineleukocyte gene expression in juvenile rats that underwent n–3 PUFAdepletion with and without energy restriction (ER). Additionally, wetested whether supplemental docosahexaenoic acid (DHA; 22:6n–3)could reverse modifications related to n–3 PUFA depletion combinedwith ER.

Methods: Male rats, 21 d old (n = 19/group), were fed ad libitum(AL) diets made with soybean oil (SO-AL), which was α-linolenicacid (ALA; 18:3n–3) sufficient, or ALA-deficient corn oil (CO-AL) for4 wk. For CO, ERwas also implemented, whereby rats were individuallypair-fed 75% against CO-AL rats with (CO-ER + DHA) and without(CO-ER) supplemental DHA (1% energy). Hematologic endpointswere determined. Peripheral leukocytes were isolated and livers werecollected. Quantitative polymerase chain reaction was performed onleukocytes measuring several genes important in lipid and energymetabolism, oxidative stress, apoptosis, and autophagy. Hepatic fattyacids (FAs) were analyzed.

Results: n–3 PUFA depletion (SO-AL vs. CO-AL) reduced expres-sion of several genes involved in the oxidative stress pathway (Nfe2l1,Nfe2l2, Gclc, Txnrd1, and Gsk3b) and energy metabolism (Thra andThrb), and the proapoptotic gene Bax. There was an additive reductionto all of these genes when animals were energy restricted (CO-ER);however, DHA supplementation (CO-ER + DHA) did not restore geneexpression back to CO-AL or SO-AL concentrations. Hepatic n–3DHAcontent was reduced in CO-AL and CO-ER compared with SO-AL,whereas DHA content in CO-ER+DHAwas elevated above all groups.


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Conclusions: The expression of oxidative stress genes in leukocytesis sensitive to n–3 PUFA depletion. However, ER proves to have a moreprofound effect on gene expression since the n–3 dependent changescould not be restored with supplemental DHA during ER.

Funding SourcesThis work was supported by USDA-ARS project 3062-51,000-053-

00D.

Maternal and Offspring Supplementation with Fish oil ImprovesMetabolic Health in Diet-Induced Obesity (P10-086)

Kalhara R Menikdiwela, Stephani Clevenger, Tochi Eboh,Boontharick Sopontammarak, London Allen, Shane Scoggin,Iurii Koboziev, Naima Moustaid-Moussa, and Latha Rama-lingam

Texas Tech University

Background: Obesity is a complex disease and a global epidemic.Half of American women of childbearing age have obesity or areoverweight. This, in turn, increases the prevalence of childhood obesityand comorbidities. Thus, early dietary interventions during pregnancymay help prevent both maternal and offspring obesity and its metaboliccomplications. Fish oil, rich in omega-3 (n–3) polyunsaturated fattyacids, exerts various health benefits, like lowering serum triglycerides,inflammation, and insulin resistance. However, whether maternalconsumption of fish oil protects offspring from the adverse effects ofmaternal obesity remains unclear.

Objectives: The aim of this study was to test the hypothesis thatfish oil (FO) supplementation during pregnancy and continuationin offspring will reduce obesity and its associated adipose tissueinflammation in the offspring.

Methods:Mice (dams) were fed a control low-fat (LF) diet, a high-fat diet (HF), or a high-fat diet supplemented with fish oil (HF-FO)for 8 wk prior to pregnancy, during pregnancy, and during lactation.After weaning,male and female offspring fromHF- orHF-FO-fed damswere either randomly assigned to the same diets (HF-HF or FO-FO) orswitched to the other diet (HF-FO or FO-HF) for 12 wk, and comparedwith the control LF diet. Physiologic outcomes included body weight,body composition, glucose, and insulin tolerance tests. Serum, adiposetissue, and other tissues were collected at the end of the feeding periodfor further analyses.

Results: Mice fed on the HF diets had significantly higher bodyweight and were insulin resistant compared with LF mice. Bothmale and female offspring who continued on the FO (FO-FO) hadsignificantly better glucose clearance and insulin sensitivity comparedwith all the HF groups. Moreover, expression of the proinflammatorymarker tumor necrosis factor α was lower in both the FO-FOand LF groups compared with the HF-HF group. Adipocyte sizewas significantly reduced (P < 0.05) in the FO groups (FO-FO,FO-HF, HF-FO) compared with the HF mice, indicating that FOsupplementation has beneficial effects on both obesity and insulinresistance.

Conclusions: FO supplementation during pregnancy and contin-uous feeding to offspring reduces adverse maternal effects of a high-fat diet on offspring. Further mechanistic studies are ongoing to better

understand the contribution of in utero high fat vs. fish oil feeding onoffspring metabolic health.

Funding SourcesAmerican Heart Association (AHA) #17AIREA33700011.

Measuring the Bioavailability of Dietary Protein with the Use ofa Dual Stable Isotope RatioMethod: A First Proof of Principle (P10-087)

Marco Mensink,1 Nikkie van der Wielen,1 NadezdaKhodorova,2,3,4 Walter Gerrits,1 Claire Gaudichon,2,3,4 andDaniel Tomé2,3,4

1Wageningen University & Research, Division of Human Nu-trition, Netherlands; 2UMR PNCA, France; 3INRA, France; and4AgroParisTech, Université Paris-Saclay, France

Background: The quality of a dietary protein depends on its aminoacid composition and bioavailability. Bioavailability can be measuredindirectly by quantifying ileal digestibility. This is, however, quite adifficult and invasive thing to do.

Objective: The objective of the present study was to test thepotential of a less-invasive, blood-meal signature dual stable isotope-based approach to evaluate bioavailability of dietary protein in humans.

Methods: On 2 occasions, 10 healthy subjects ingested a puddingdivided into 9 portions every 20min. The isocaloric puddings containedeither 50 (“high”) or 25 g (“low”) of intrinsically labeled [15N]milkprotein (2% enriched), and both contained 400 mg of intrinsicallylabeled [13C]spirulina (99% enriched). Isotopic enrichment in the mealprotein and in the total serum free amino acid (AA) fraction wasdetermined at baseline and in the postprandial period by elementalanalysis-isotopic ratio mass spectrometry. The study was executed in2 different parts (n = 4 and n = 6).

Results: The high-protein meal resulted in an ∼2-fold higher 15Nenrichment in serum free AA compared with the low-protein meal,whereas the postprandial serum AA 13C enrichment was the same forboth meals (see Figure 1). Unexpectedly, the 13C enrichment appearedto be almost 2-fold lower in the second part of the study for bothconditions, whereas the 15N enrichment was comparable.

The blood-to-meal ratio of the isotopic signature, i.e. (15N:13Cratio—serum)/(15N:13C ratio—meal), reflects the relative digestibility ofthe 2 proteins. The relative digestibility was comparable between thehigh- and low-protein meals, but, again, differed between parts 1 and 2.Assuming a digestibility of milk protein of 95%, the protein digestibilityof spirulina was calculated to be 75.1% ± 28.9% and 74.6% + 19.3%for the high- and low-protein conditions for part 1 of the study and36.5% + 19.6% and 37.4% + 9.8% for part 2.

Conclusions: A clear postprandial response of 13C and 15Nenrichment in the total serum free AA was observed. These values,together with the enrichment levels in the meal, allowed the calculationof the relative digestibility of milk protein and spirulina, which was notaffected by meal composition. Unexpectedly, a difference in serum AA13C enrichment, not 15N, was seen between the 2 parts of our study. Thisclearly deserves further attention.

Funding SourcesDutch DairyAssociation (NZO).


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FIGURE 1 Serum free AA enrichment after frequent ingestion of a meal from t = 0 to t = 160 min. Enrichment is expressed in APEcompared with baseline enrichment. APE, atom percentage excess.

Partial Replacement of Dietary L-Methionine by DL-2-Hydroxy-4-methylthiobutyrate Decreases Transsulfuration and Red BloodCell Reduced Glutathione Fractional Synthesis Rate in Piglets(P10-088)

Cornelia C Metges,1 Ilka Rasch,1 Solvig Goers,1 ArminTuchscherer,1 Behnam Saremi,2 John Htoo,2 and Bjoern Kuhla1

1Leibniz Institute for Farm Animal Biology (FBN), Germany; and2Evonik Nutrition & Care GmbH, Germany

Background: Cysteine (Cys) is synthesized from methionine (Met)via trans-sulfuration (TS). It is the rate-limiting amino acid for thesynthesis of reduced glutathione (GSH). GSH is a component of theantioxidant defense system. DL-2-Hydroxy-methylthiobutyrate (DL-HMTB) is a Met analog, and it has been suggested to increase GSHsynthesis when it partially replaces dietary Met.

Objective: The aim of this study was to determine methyl (Qm) andCys fluxes (Cysflux), transmethylation (TM) and TS rates, as well as redblood cell (RBC) GSH synthesis in a piglet model fed a diet in whichL-Met was partly replaced by DL-HMTB.We hypothesized that dietaryreplacement of L-Met by DL-HMTB alters Qm, Cysflux, TM, TS, andGSH synthesis rates.

Methods: A total of 31 male (German Landrace) 29-d-old weanedpiglets were allocated to a diet meeting 70% of the recommended dailyamount of Met and Cys and supplemented with 30% of molar Metequivalents as either L-Met (n = 12), DL-Met (n = 10), or DL-HMTB(n = 9). The piglets were equipped with jugular and carotid catheters,and received at age 47 d a primed 9-h intravenous infusion of L-[1-13C-methyl-2H3]Met and L-[3,3-2H2]Cys for 3 h in a fasted condition,followed by feeding half-hourly on small meals for 6 h. At age 57 d,fasted piglets were intravenously infused with [2H2]glycine for 8 h.Plasma Met and Cys enrichments were determined by GC-MS, andquantitative kinetics (expressed as µmol · kg–1 · h–1) were calculated.The [2H2]glycine enrichment in red blood cell (RBC)GSHwas analyzedby GC-MS to calculate the GSH fractional synthesis rate (FSR, %/d).Concentrations of RBC GSH and oxidized glutathione (GSSG), and thesum of GSH and GSSG (total GSH, mM), were quantified by HPLC.The differences between diets were analyzed by ANOVA using PROCMIXED of SAS.

Results: Fed state whole-body Qm (140 ± 2.4), Cysflux (99 ± 2.4),and TM (46 ± 1.8) did not differ between groups (P > 0.05). TS (19 vs.28) and theTS:TMratio (0.4 vs. 0.6)were lower inDL-HMTBcomparedwith L-Met (P < 0.05). TS (19 vs. 28) and the TS:TM ratio (0.4 vs. 0.6)were lower in DL-HMTB compared with L-Met (P < 0.05) but not DL-Met. GSH FSR (34 vs. 64) and total RBC GSH concentrations (1.7 vs.


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1.9) were lower (P < 0.05) in DL-HMTB compared with L-Met, andDL-Met was intermediate. The L-Met andDL-Met groups did not differ.

Conclusions:ADL-HMTB compared with an L-Met supplementeddiet reduced TS, RBC GSH FSR, and total GSH concentrations.

Funding SourcesInstitutional budget to the Leibniz Institute for FarmAnimal Biology

(FBN) and Evonik Nutrition & Care GmbH.

Postprandial Lipemic Responses to a Meal Rich in Interesterifiedor Noninteresterified Palm Stearin/Palm Kernel Fat Blends: ARandomized Controlled Trial in Older Adults (P10-089)

Sarah Berry, Charlotte Mills, Robert M Gray, and Wendy L Hall

King’s College London

Background: The 2003 WHO/FAO technical report on diet,nutrition, and the prevention of chronic diseases stated that the intakeof industrially produced trans-fat should be as low as possible owingto strong evidence demonstrating harmful health effects. Interesterified(IE) fats rich in palmitic acid (produced from palm oil fractions,yielding semisolid fat) are used as an alternative, but health impacts ofthe most commonly consumed IE fat blends are unknown.

Objective: The primary objective of this study was to investigate theimpact of IE fat on postprandial lipemia (PPL), an independent riskfactor for cardiovascular disease.

Methods: We undertook a 3-armed, double-blind, randomizedcontrolled trial in healthy males and females (n = 20), aged 45–75 y,

FIGURE 1 Change from baseline in plasma triacylglycerol over the 8-h study period (n = 20, mean ± SEM).

to compare effects of single test meals (muffin with a milkshake; 897kcal, 50 g of fat, 16 g of protein, 88 g of carbohydrate) on postprandialplasma triacylglycerol (TAG) concentrations. Test fats were IE 80:20palm stearin:palm kernel (PS:PK) fat and the equivalent non-IE fat,which were compared with a control high-MUFA oil known to induce apronounced lipemic response (rapeseed oil, RO). Bloodwas collected atbaseline, and hourly thereafter for 8 h. Data were log transformedwherenot normally distributed. Statistical analysis was performed using linearmixedmodeling, adjusting for baseline values with post hoc adjustmentfor multiple comparisons and chi square test for Tmax. The incrementalarea under the curve (iAUC) was calculated using the trapezoidmethod.

Results: There was a significant time × treatment interaction(P = 0.006) in change from baseline of plasma TAG concentrations(Figure 1), with significant differences between IE fat and RO and non-IE fat and RO (both P = 0.001), but not between IE fat and non-IE fat(P= 0.407). There was no effect of treatment for iAUC0–4 h or iAUC0–8 h

(P= 0.219 andP= 0.107, respectively). Norwere there treatment effectsfor Cmax or Tmax (P = 0.198 and P = 0.571, respectively).

Conclusions: The process of interesterification of an 80:20 PS:PKblend does not impact on PPL in older, healthy adults. However, otherpostprandial markers yet to be assessed in this trial will help us tofully understand the impact of consuming these fats on metabolicparameters.

Funding SourcesBBSRC DRINC.


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Daily Exercise Combined with a High-Protein Diet Promotes FatLoss and Improves Fatty Acid Oxidation during Short-Term CaloricRestriction (P10-090)

Chaise AMurphy, Elly Glazier, Jay Petersen, andKarsten Koehler

University of Nebraska—Lincoln

Background: Effective weight loss requires targeted loss of fat mass(FM). Although both caloric restriction (CR) and exercise can reduceadiposity, the most effective strategies to maximize fat loss remainunclear.

Objectives: The objective of this study was to determine the abilityof a high-protein diet (HPD) to favorably influence body compositionand fatty acid oxidation (FAO) over 5 d of CR in healthy, active men.The HPD was hypothesized to improve retention of lean mass (LM),resulting in greater loss in FM and greater FAO both at rest and duringexercise.

Methods: In this randomized, crossover trial, participants under-went 3 conditions: CR with low protein intake [CRLP; 0.8 g/kg bodyweight (BW)], CR with high protein intake (CRHP; 1.7 g/kg BW),and control (CON; 1.7 g/kg BW). CR was achieved by reducingenergy availability to 15 kcal/kg fat-free mass (FFM), whereas energyavailability was maintained at 40 kcal/kg FFM in CON. Throughoutall conditions, participants completed supervised exercise sessions ona bicycle ergometer set to expend 15 kcal/kg FFM. Before and aftereach 5-d condition, body composition was assessed via bioimpedance,and FAO and peak oxygen uptake (VO2peak) were measured during anincremental exercise test.

Results: Body weight decreased (P < 0.001) in CRLP (–1.97 ±0.22 kg) and CRHP (–1.80 ± 0.52 kg), but remained stable in CON.Loss of FM was significant in CRHP (–1.14 ± 0.23 kg, P < 0.05),but not in CRLP (–0.57 ± 0.48 kg), whereas the reduction in FFMwas only significant in CRLP (–1.4 ± 0.59 kg, P < 0.01), not inCRHP (–0.6 ± 0.57 kg). Maximal rates of FAO increased (P < 0.01)in CRHP (+ 0.22 ± 0.04) and CRLP (+ 0.15 ± 0.06), but not inCON. VO2peak increased only in CRHP (+ 2.66 ± 0.74 mL/kg/min,P < 0.05), not in CRLP (+ 1.00 ± 0.75 mL/kg/min) or CON (–0.46 ±1.41 mL/kg/min).

Conclusions:Despite similar overall weight loss, short-term caloricrestriction combined with exercise resulted only in significant fat losswhen participants were provided with a high-protein diet. Althoughcaloric restriction improved maximal fat oxidation independently ofprotein intake, only a high-protein diet appeared to provide modestperformance improvements during short-term calorie-restrictedweight loss.

Funding SourcesUniversity of Nebraska Foundation and USDA National Institute of

Food and Agriculture.

Improvement Effect of Egg White Hydrolysate on GlucoseTolerance in Mice (P10-092)

Masaru Ochiai

School of Veterinary Medicine, Kitasato University, Japan

Background: Egg white (EW) contains high protein and low fat,and is commonly used in food processing. However, it is also high inallergenic proteins. In order to prevent the risk of onset of allergy forhumans, protein hydrolysis methods have recently been used. SomeEWhydrolysates (EWH) have been reported to suppress oxidative stressand improve fat metabolism. However, improvement effects of EWHonglucose metabolism have not yet observed. From our previous studies,dietary EWH improved fasting plasma glucose concentration andglucose intolerance in type 2 diabetic animals, but their mechanismshave not yet been clarified.

Objective:The aim of this study was to investigate the dietary effectsof EW and EWH on glucose tolerance in different ways to clarify theeffect of EW and EWH on intestinal glucose absorption in mice.

Methods: Type 2 diabetic NSY mice, aged 4 wk, were divided into 4dietary groups: a normal AIN-93G diet group (N); a high-fat and high-sucrose (HFS) casein-based diet group (NH); an HFS EW-based dietgroup (NE); and an HFS EWH-based diet group (NEH). Mice werefed their respective diets for 8 wk. At the end of weeks 6 and 7, anoral glucose tolerance test (OGTT) and insulin tolerance test (ITT)were respectively conducted in the first study. In the second study, anintraperitoneal glucose tolerance test (ipGTT)was conducted at the endof week 7. At the end of the treatment period, biochemical parameterswere measured to clarify the mechanisms involved.

Results: In the first study, the plasma glucose concentration wassuppressed in theNE group during bothOGTTand ITT and suppressedin the NEH group during OGTT, but not during ITT. In second study,the plasma glucose concentration was similarly suppressed in the NEHgroup during ipGTT, but the suppressive effect wasweakened comparedwith OGTT. The final plasma insulin concentration was lower in theNE and NEH groups in both studies. The level of total glucagon-likepeptide-1, which is closely related to insulin secretion from pancreasβ cells, was not different between the groups in both studies. Fecaltriacylglycerol excretion was increased in the NE and NEH groups,whereas liver triacylglycerol content was suppressed in the NE group.

Conclusions: In addition to improving fat metabolism, EWHimproves glucose tolerance, mainly via a small intestine–relatedmechanism in mice.


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FIGURE 1

Branched-Chain Amino Acids and Lipid Profile among NormalWeight Adults with Impaired Glucose Tolerance (P10-093)

Akinkunmmi Paul Okekunle, Liyan Liu, Rennan Feng, Ying Li,and Chang-Hao Sun

Harbin Medical University, China

Objectives: This study explored the relation between serumbranched-chain amino acids (BCAAs) and lipid profiles among normal-weight adults with impaired glucose tolerance (IGT).

Methods: Demography, anthropometry, and blood pressure (BP)measurements, and fasting and 2-h blood samples were obtained from15 newly onset IGT and 15 normoglycemic subjects [(matched for age,sex, and body mass index (BMI)] using standardized methods. Fastingblood glucose (FBG), 2-h plasma glucose (2h-PG), fasting insulin (FI),total cholesterol (TC), triglycerides (TG), HDL cholesterol, and LDLcholesterol concentrations were determined using an autoanalyzer withcommercial kits, and BCAAs (isoleucine, leucine, and valine) wereprofiled using tandem mass spectrometry. A χ ² test or t test, forcategorical (%) or continuous variables (means± SD), respectively, was

applied to compare differences between normal weight without IGT(NW) and normal weight with IGT (NWI) groups. Pearson’s correlation(r) was conducted to evaluate the relations between BCAAs, lipidprofiles, FBG, 2h-PG, and FI, and 2-sided P < 0.05 was consideredstatistically significant.

Results: Age, waist circumference, systolic BP, and BMI weresignificantly higher in the NWI group. Similarly, FBG and 2h-PG weresignificantly (P < 0.01) higher in the NWI group (5.2 ± 1.3 and10.4 ± 4.5 mmol/L, respectively) than in the NW group (3.9 ± 0.3 and5.4 ± 1.0 mmol/L, respectively), but FI differed insignificantly. Thoughserum LDL cholesterol tended to be higher in the NWI group (3.4± 0.8mmol/L) than in the NW group (2.8 ± 0.8mmol/L), TC, TG, HDLcholesterol, and the BCAA profiles were all insignificantly differentbetween the 2 groups. Correlation coefficients between FBG, 2h-PG,FI, and BCAA were insignificantly different (even after stratifying theIGT status). However, HDL cholesterol was negatively correlated withIle (r = –0.37; P < 0.05) and Leu (r = –0.37; P < 0.05).

Conclusion: A higher concentration of BCAAs negatively alters theHDL cholesterol profile.


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Funding SourcesThe China Scholarship Council (2015BSZ778) and the National

12th Five-Year Scientific and Technical Support Program of China(2012BAI02B02).

Do Saturated Fatty Acids Truly Activate Inflammation? Experi-mental Confounders and Context Specificity (P10-094)

Kikumi D On-Moore, Michael Blackburn, and Sean H Adams

Arkansas Children’s Nutrition Center

Background: Excess dietary saturated fat consumption has beenlinked to promotion or exacerbation of cardiometabolic diseases andother conditions associated with chronic low-grade inflammation.Based primarily on cell culture experiments, saturated fatty acids (SFAs)are proposed to promote inflammation and contribute to metabolicdysfunction through toll-like receptor activation. Studies are oftencomplicated by a requirement for carriers [e.g., bovine serum albumin(BSA)] or solvents (e.g., ethanol) to increase SFA solubility, yet BSA canitself induce inflammation and ethanol is a cytotoxin.

Objective: The aim of this study was to ascertain if these factorsinfluence SFA-associated inflammation, which we did by measuringresponses of RAW264.7 monocytes or macrophages and C2C12myotubes to various BSAs, ethanol and cyclodextrin (an alternative toBSA).

Methods: The inflammatory responses of RAW264.7 macrophagesand C2C12 myotubes were determined in response to various lots andconcentrations of commercially available low-endotoxin, fatty acid–free BSA (0.33–2% wt:vol), multiple concentrations of ethanol (0.06–1% vol:vol), and cyclodextrins (0.3–6.0 mM). The cellular response topalmitic acid (PA) (200–600 µM) was assessed.

Results: Low-endotoxin BSA preparations activated, whereas 0.5–1.0% ethanol inhibited, RAW264.7 tumor necrosis factor (TNF)-αrelease. Ethanol modestly increased interleukin (IL)-6 secretion inC2C12 myotubes. Cyclodextrins were tested as an alternative carrier ofPA, but their usefulness in cell culture was limited owing to toxicityand solubility issues. Using a lower-inflammation lot of BSA and noethanol, ∼24 h of sodium PA treatment (up to 600 µM) failed totrigger RAW264.7 TNF-α release, and in fact significantly dampenedBSA-induced inflammation by >50%. In C2C12 myotubes, only highPA concentrations (500–600 µM) elicited IL-6 secretion (>2.5-foldincrease); acute PA (200 or 500 µM) did not activate mitogen-activatedprotein kinase pathways above that of fresh media alone.

Conclusion:These results highlight the importance of experimentalconditions in studies exploring SFA inflammation effects. The limited(or even anti-inflammatory) effects of PA that we observed indicatethat immunomodulatory effects of SFAs are context specific. Thus,caution is needed when interpreting the literature related to putativeproinflammatory effects of SFA.

Funding SourcesARS 6026-51,000-010-05S.

Detection of Plasmalogen Hydroperoxide in Human Plasma(P10-095)

Yurika Otoki,1 Shunji Kato,2 Teruo Miyazawa,1 and KiyotakaNakagawa1

1Tohoku University, Japan; and 2Tokai University School ofMedicine, Japan

Background: Plasmalogen (Pls) is a physiologically important classof vinyl ether–linked phospholipid. Recently, patients suffering fromneurodegenerative disorders have been reported to exhibit reducedconcentrations of choline Pls (PC-Pls) and ethanolamine Pls (PE-Pls) inblood plasma. One of the potential cause of this reduction is oxidation,since Pls is vulnerable to oxidation owing to the presence of the vinylether bond. So far, Pls oxidation has been mainly discussed with regardto cleavage of the vinyl ether bond (i.e., yielding a lysophospholipid),despite the fact that the polyunsaturated fatty acid (PUFA) moiety ofPls is known to be highly susceptible to oxidation, owing to the lack ofa suitable analytical method.

Objectives: In this study, we aimed to construct a novel method fordetection of Pls bearing oxidized PUFA and reveal whether the PUFAmoiety of Pls is oxidized.

Methods: Standard PC-Pls (16:0/18:2) was oxidized by 15-lipoxygenase. The resultant PC-Pls, bearing 13-hydroperoxy-octadecadienoyl at sn-2 (PC-Pls 16:0/13-HpODE), was analyzed bytandemmass spectrometry (MS/MS) in the presence of the sodium ion.A hydroperoxide-specific product ion was used for high-performanceLC-MS/MS operated in multiple reaction monitoring mode. Theplasma phospholipid fraction was then analyzed.

Results: In the presence of sodium ion, the characteristic production from PC-Pls 16:0/13-HpODE was determined. Under the optimalLC-MS/MS conditions, PC-Pls 16:0/13-HpODEwas detected in humanplasma. It became clear that the PUFAmoiety of Pls was also a target foroxidation.

Conclusion: We are now applying the novel method to analyzeclinical samples. The method described here would provide a betterunderstanding the physiologic role of Pls in vivo.

Funding SourcesApplication Procedures for Grants-in-Aid for Scientific Research—

KAKENHI.

EPA-Derived Mediators Are Lowered in Murine Obesity andRescued with Dietary Intervention to Improve Glucose Clearance(P10-096)

Anandita Pal,1 William Guesdon,2 Adam J Amorese,2 MariaJJ Torres,2 P Darrell Neufer,3 Espen Spangenburg,2 MichaelArmstrong,4 Nichole Reisdorph,4 and Saame R Shaikh5

1University of North Carolina at Chapel Hill; 2East CarolinaUniversity; 3Brody School of Medicine, East Carolina Diabetes andObesity Institute, East Carolina University; 4University of Colorado-Denver; and 5Gillings School of Global Public Health and School ofMedicine, University of North Carolina at Chapel Hill

Objectives:Diet-induced obesity impairsmetabolic and physiologicoutcomes through complex mechanisms. We explored the role of lipidmediators synthesized from polyunsaturated fatty acids (PUFA) inregulating glucose and insulin sensitivity.

Methods: We first conducted mass spectrometry analyses ofspecific tissues from mice consuming a high-fat diet, with a focus onpotential lipid mediators synthesized from n–6 and n–3 PUFAs. Wesubsequently assessed the potential of eicosapentaenoic acid (EPA) and


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its metabolites in improving glucose and insulin sensitivity in miceconsuming a high-fat diet. We fed 8-wk-old mice with a high-fat dietor a high-fat diet supplemented with 12-HEPE, 18-HEPE, or EPA for15 wk and compared them with mice on a normal chow diet (CD).

Results: Select arachidonic acid derivativeswere elevated in epididy-mal adipose tissue and liver of obese mice compared with lean controls.Notably, the hydroxylated 12-HEPE and 18-HEPE, synthesized fromEPA, were dramatically lowered in the adipose tissue and liver of theobese mice. In contrast, 14-HDHA and 17-HDHA, synthesized fromdocosahexaenoic acid, were not influenced by the high-fat diet relativeto the controls. Based on these results, we tested the effects of short-term administration of 12-HEPE and 18-HEPE (4 d) in addition tolong-term administration with EPA (15 wk) on glucose clearance andfasting insulin concentrations. EPA, and to some extent 18-HEPE,improved fasting glucose concentrations and the homeostatic modelassessment insulin resistance index. In addition, 18-HEPE increased invitro force production of the skeletal muscle without affecting musclesize, suggesting an improvement in muscle quality.

Conclusions: Taken together, these results suggest that EPA ethylesters, potentially by triggering production of 18-HEPE (a pathwaymetabolite of resolvin E1), increase insulin-glucose sensitivity and forceproduction in skeletal muscle.

Funding SourcesThe research was supported by NIH R01AT008375 (to SRS), NIH

R01 AR06666 (to ES).

The Effect of Dietary Cholesterol on High-Density LipoproteinCholesterol Levels inMen andWomen: AMeta-analysis of Random-ized Controlled Trials (P10-097)

Orsolya M Palacios,1 Melissa Vincent,2 Bruce Allen,3 LynneHaber,2 and Kevin Maki1

1Midwest Biomedical Research/Center for Metabolic and Cardio-vascular Health, MO; 2University of Cincinnati; and 3IndependentConsultant, Chapel Hill, NC

Background: Intakes of dietary cholesterol and dietary fatty acidsaffect circulating cholesterol concentrations, including the concentra-tion of HDL cholesterol, but the impact of dietary cholesterol on HDLcholesterol, after adjustment for dietary fats intake, remains uncertain.

Objective: The goal of this study was to perform a meta-regression analysis, using data from randomized dietary interven-tion trials, to assess the relation between the change in dietarycholesterol and the change in HDL cholesterol, while controlling forintakes of saturated, trans, polyunsaturated, and monounsaturatedfatty acids using the coefficients from the Mensink and Katanequation (2003).

Methods: A literature search (1965–2016) identified 3616 abstracts,of which 51 publications met all inclusion criteria and no exclusioncriteria. Data from the 51 publications were extracted and used forthe Bayesian meta-regression analysis. Two different biological modelshapes were examined: linear and the nonlinear Michaelis-Menten(MM).An additional parameterwas considered in themodel tomediatethe effect of baseline cholesterol intake.

Results: No significant relation was found between the changein dietary cholesterol intake and the change in HDL cholesterolconcentrations in pooled data frommen andwomen.However, both the

linear and MM models indicate that the change in dietary cholesterolintake is inversely related to the change in circulating HDL cholesterolconcentration in men, but positively related to the change in circulatingHDL cholesterol in women. In men, mean predicted changes in HDLcholesterol for an increase of 100mg/d of dietary cholesterol were –0.30and –1.44 mg/dL for the linear and MMmodel, respectively, and for anincrease of 300 mg/d, the corresponding mean predicted values were –0.90 and –2.05 mg/dL, respectively. In women, mean predicted changesin HDL cholesterol for an increase of 100 mg/d of dietary cholesterolwere 0.50 and 1.61 mg/dL for the linear and MM model, respectively,and for an increase of 300 mg/d, the corresponding mean predictedvalues were 1.50 and 2.29 mg/dL, respectively.

Conclusions: These results suggest a possible interaction betweensex and the HDL cholesterol response to dietary cholesterol. However,the clinical implications of changes in HDL cholesterol associated withincreased dietary cholesterol intake remain uncertain.

Funding SourcesFunded by the American Egg Board-Egg Nutrition Center, Park

Ridge, IL.

Effects of Intermittent Energy Restriction with a MediterraneanDiet versus the Dietary Approaches to Stop Hypertension Diet onOverall and Visceral Adiposity: A Pilot Study (P10-098)

Chloe Elizabeth Panizza,1 Unlee Lim,1 KimMYonemori,1 KevinDCassel,1 Loic LeMarchand,1 LynneWilkens,1 Carol J Boushey,1Yurii B Shvetsov,1 and Michelle Harvie2

1University of Hawaii Cancer Center; and 2Manchester UniversityHospital Foundation NHS Trust, United Kingdom

Objectives: The aims of this study were to develop a protocol,and assess adherence and the short-term effects of intermittent energyrestriction with a Mediterranean (IER + MED) diet vs. the DietaryApproaches to Stop Hypertension (DASH) diet on overall and visceraladiposity measures in East Asian adults.

Methods: Men and women in Hawaii (n = 60), aged 35–55 y,with a body mass index of 25–40 kg/m2, a visceral fat area (VFAT)≥90 cm2 for men and ≥80 cm2 for women, of Japanese, Chinese, orKorean ancestry, were randomized equally to the IER+MED or DASHdiet for 12 wk. The IER + MED group was instructed to restrict 70%energy (34%, 33% and 33% distribution of protein, carbohydrate, andfat, respectively) on 2 d and follow the MED diet (25%, 45%, 30%)and meet their estimated energy requirement (EER) for the other 5days each week. The DASH group was instructed to follow the DASHdiet (20%, 53%, 30%) and meet their EER. Dietitians provided initialinstruction for the diets and contacted participants 7 times, primarilyby telephone, for support and compliance assessment. At baseline andat weeks 6 and 11, 4-d mobile food records were completed. Trainedtechnicians measured anthropometric parameters and dual-energyX-ray absorptiometry-based body composition, including estimatedVFAT at L4-L5, at baseline and week 12.

Results: Analyzing the 54 participants who completed the study(IER + MED, n = 26; DASH, n = 28), the IER + MED group reportedcompleting 90% of their restricted diet days by week 12. At week 11,the IER + MED group had a 60% energy deficit on the restricted days(30%, 28%, 44%) and a 50% energy deficit (25%, 37%, 39%) on the non-restricted days. The DASH group had a 30% energy deficit across all


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days (19%, 45%, 37%).Atweek 12, participants in the IER+MEDgrouphad lost greater amounts of body weight (5.9 ± 3.1 vs. 3.2 ± 3.4 kg inDASH;P= 0.006), waist circumference (WC) (7.0± 4.5 vs. 4.4± 3.1 cminDASH;P= 0.018), andVFAT (1.3± 29.5 cm2 reduction vs. 4.2± 20.5cm2 increase in DASH; P = 0.43].

Conclusion: In this short-term pilot study, the IER + MED grouphad greater reduction in weight and WC than the DASH group. Thelarger effect of the IER+MEDdiet on adipositymeasuresmay be owingto the IER, larger energy deficit, and/or different diet composition. AnIER + MED vs. DASH intervention can be implemented in an EastAsian adult population in Hawaii.

Funding SourcesThe William & Ellen Melohn Endowed Research Fund, University

of Hawaii. Supported by the National Cancer Institute at the NIH P30CA071789, P01CA168530, U01CA164973.

Effect ofα-LinolenicAcid, EPA, orDHACombinedwithEstrogenon Cholesterol Metabolism in Ovariectomized Rats (P10-099)

Yongsoon Park,1 Chung Park,1 Soomin Yu,1 and Youri Jin2

1Hanyang University, South Korea; and 2Hanyang University Hos-pital, South Korea

Objectives: The purpose of the study was to determine the hypoc-holesterolemic mechanism of α-linolenic acid (ALA), eicosapentaenoicacid (EPA), and docosahexaenoic acid (DHA) individually with 17β-estradiol-3-benzoate (E) in ovariectomized (OVX) rats.

Methods: Female rats were given 1 of 4 diets containing 0% n–3polyunsaturated fatty acids, 1% ALA, 1% EPA, or 1% DHA of the totalenergy intake for 12 wk. At week 8, rats were sham-operated or OVX,and during the last 3 wk of experimental period the OVX rats wereinjected with corn oil vehicle or E every 4 d to mimic the menstrualcycle.

Results: Rats fed EPA or DHA with E showed a significant decreasein the serum concentrations of triglyceride and LDL cholesterol, andan increase in the serum concentration of HDL cholesterol. Supple-mentation of EPA or DHA significantly increased expression of thephosphorylated adenosine monophosphate–activated protein kinase(p-AMPK) to AMPK ratio, cholesterol 7α-hydroxylase (CYP7A1),sterol 12α-hydroxylase (CYP8B1), and sterol 27-hydroxylase, anddecreased the expressions of 3-hydroxy-3-methylglutaryl coenzyme Areductase, sterol regulatory element-binding protein-2, and proproteinconvertase subtilisin/kexin type 9 in liver. However, E significantlydecreased the hepatic expressions of CYP7A1 andCYP8B1. In addition,E significantly increased the hepatic expressions of estrogen receptor-αandβ . Supplementation ofALAhad no significant hypocholesterolemiceffect and did not changed the hepatic protein expression related withcholesterol synthesis and clearance.

Conclusions: The present study showed that the hypocholes-terolemic effect of EPA or DHA was derived from increased hepaticbile acid synthesis, upregulated LDL-receptor, and decreased hepaticcholesterol synthesis. However, the hypocholesterolemic effect of Ewas derived from decreased hepatic cholesterol synthesis and up-regulated LDL-receptor, but not by the decreased hepatic bile acidsynthesis.

Funding SourcesThis research was supported by Basic Science Research Program

through the National Research Foundation of Korea (NRF) funded bythe Ministry of Education (NRF-2015R1D1A1A09060823).

Influence of Meal Caloric Distribution in Metabolic SyndromeParameters among College Students (P10-100)

Ashley Petitta, Alexis Elinkowski, and David Aguilar-Alvarez

Weber State University, UT

Objectives: The aim of this study was to investigate the influenceof meal calorie distribution on metabolic syndrome (MetS) parametersin college students. We hypothesize that variance in the percentage ofcalories eaten at each meal will affect MetS parameters in both malesand females.

Methods:We assessedMetS parameters in 168 student participants,aged 18–54 y. From each participant, we collected 2-d diet recordsand analyzed them using Diet and Wellness Plus. Participants wereseparated by gender (male, n= 53; female, n= 115) and bymeal caloriedistribution. Groups included high, medium, and low percentage ofcalories in breakfast, lunch, dinner, and snacks. ANOVA tests were usedto determine themean differences inMetS parameters between themealcalorie distribution groups.

Results: Our results show that women in the high calorie breakfastdistribution group presented lower systolic blood pressure than womenin the low or medium breakfast group (μ = 107.3, 113.3, and115.01mmHg, respectively, P≤ 0.05).Men in the high snack consumergroup presented higher HDL cholesterol (μ = 41.3, 35.1, and 32.3mg/dL, respectively, P≤ 0.01) as well as higher blood glucose (μ = 97.8,92.1, and 91.5mg/dL, respectively, P≤ 0.05) than the low andmoderatesnack groups.

Conclusion: In accordance with our results observed in the femalesubgroup, previous studies have shown that skipping breakfast increasescortisol concentrations, which may result in higher blood pressure.Furthermore, many breakfast-associated foods have been shown toexert blood pressure–lowering effects. The differences observed inmale HDL cholesterol may be owing to HDL cholesterol–promotingfoods consumed as snacks. High snack consumption is linked toincreased exercise, which is known to increase HDL cholesterol. Thisstudy suggests that meal calorie distribution affects MetS parametersdifferently between genders.

An Obesity Intervention Emphasizing Balanced Higher Proteinfrom Red Meat Sources Reduces Cardiometabolic Risk (P10-101)

Kathryn N Porter Starr,1,2 Margery Connelly,3 ShelleyMcDonald,1 Richard J Sloane,1 Alyssa D Podwika,4 KimHuffman,1 William Kraus,1 and Connie W Bales1,2

1Duke University School of Medicine, NC; 2Durham VA MedicalCenter, NC; 3LabCorp; and 4Meredith College, NC

Objective: The recognized benefits of generous, balanced intakesof high-quality protein to muscle mass and strength in older adultsis tempered by concerns about the cardiometabolic impact of dietarysources of animal protein. The objective of this study was to explorethe cardiometabolic impact of a protein-enhanced (predominantly lean


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beef and pork) versus an RDA-level protein weight reduction diet inobese middle-aged and older women and men.

Methods: Obese participants (mean body mass index: 37.3 kg/m2;90% female), aged ≥45 y (mean = 64 y), were randomly assigned toeither an RDA-level protein Control (0.8 g protein/kg body weight)or a balanced, higher protein (1.2 g protein/kg body weight; 30 g ofhigh-quality protein per meal) weight loss diet (500 kcal deficit) for6 mo. Participants in the Protein group were provided with proteinas lean and very lean red meat for 2 of 3 meals daily. Blood samplescollected at baseline and study completion were analyzed using nuclearmagnetic resonance spectroscopy to quantify cardiometabolic riskfactors: lipids, lipoprotein size and number, glycoprotein acetylation(GlycA), trimethylamine-N-oxide (TMAO), and lipoprotein insulinresistance index (LP-IR) scores.

Results: Those participants who completed the study in both theControl (n = 27) and Protein (n = 53) groups lost weight (6–7%;P< 0.0001) and had improved function (6-min walk; P< 0.0001); theyalso had reduced fasting triglycerides (P< 0.02), large triglyceride-richlipoprotein particles (P= 0.02), andplasma glucose (P< 0.003).Neithergroup experienced a change in total cholesterol, HDL cholesterol,LDL cholesterol, apolipoprotein A-I, apolipoprotein B, or TMAO (allP> 0.05). GlycA concentrations were reduced in the Control (P= 0.04)but not in the Protein (P > 0.05) group. LDL particle number (LDL-P)and LP-IR scores were reduced in the Protein (P = 0.04 and 0.0004,respectively) but not in the Control (P > 0.05) group.

Conclusions:Although further study in larger samples is warranted,these findings support the conclusion that a balanced, higher-protein(predominantly as lean red meat) hypocaloric diet not only leads toweight loss and improved function, but may also benefit insulin resis-tance (LP-IR) and lessen cardiovascular risk (LDL-P) in obese middle-aged and older adults. These findings have important implications fortreatment of sarcopenic obesity and alignwith recent recommendationsthat older adults consume generous, balanced amounts of high-qualityprotein throughout the day.

Funding SourcesThis study was funded by the Beef Checkoff and Pork Checkoff,

and received additional support from the North Carolina Pork Council,Smithfield Foods, NIH (5T32 AG000029), and the US Departmentof Veterans Affairs Rehabilitation Research and Development ServiceProgram (CDA-2/IK2 RX002348).

Grain SorghumLipidProtects againstAtherogenicDiet–InducedChanges in Energy Metabolism and Short-Chain Fatty Acid Profilein a Hamster Model (P10-102)

HaowenQiu, Sami Althwab, Nguyen TienAn, andVicki Schlegel


Background: Grain sorghum (GS) lipid extract has shown protec-tion against high cholesterol in both plasma and liver, and, to a lesserdegree, remediation in the gut microbiota in response to an atherogenicdiet. However, the impact of GS lipids on cellular stress induced by sucha diet remains largely unknown.

Objective: The objective of this research was to determine theability of GS crude lipid (GS-CL), obtained from the surface of GSwhole kernel, to protect energy metabolism and the short-chain fatty

acid (SCFA) profile produced by the gut microbiome, which may benegatively affected by an atherogenic diet.

Methods:Male hamsters were fed with either a low-fat control diet,an atherogenic diet, or an atherogenic diet supplemented with 1, 3, or5% (wt:wt) GS-CL for 4 wk. Colon tissue was collected after euthanasiaand analyzed by targeted metabolomics.

Results: Analysis showed that the atherogenic diet disrupted thecellular redox balance and impaired energy generation, promotinggluconeogenesis and anaplerosis from pyruvate and amino acids. TheGS-CL supplements were able to maintain energy levels and mitigatedthe accumulation of glycolysis and tricarboxylic acid intermediates, butwere not able to rebalance the redox equilibrium completely. More-over, multivariate analysis, including orthogonal partial least squaresdiscrimination analysis, demonstrated that the GS-CL supplementswere able to gradually shift the atherogenic affected metabolomics tolevels trending or comparable to the control level. In addition, the GS-CL supplements positively affected the SCFA produced by the colonmicrobiome against atherogenic diet–induced changes.

Conclusions: This study provides whole-scale impact of GS-CLon cellular energy and central carbon metabolism using a combinedapproach of metabolomics and multivariant analysis, which effectivelydemonstrate the potentials of GS-CL as a supplement to prevent orcontain chronic metabolic diseases.

Funding SourcesFunding for this research is provided by Nebraska Grain Sorghum

Board.

Fatty Acid, Retinol, and Carotenoid Content of Eggs Differ withHen Strain, Age, and Rearing Environment (P10-103)

Susan K Raatz,1 Laura Heflin-Morgan,2 Ramon D Malheiros,3Kenneth Anderson,3 Michael Bukowski,1 LuAnn Johnson,1 andKeith Voller1

1USDA; 2USDA, Agricultural Research Service, Grand Forks Hu-man Nutrition Research Center; and 3North Carolina State University

Objective: The objective of the current study was to determine theeffect of rearing system, strain, and hen age on fatty acid and carotenoidcontent of whole raw egg. We hypothesized that concentrations ofunsaturated fatty acids and carotenoids are higher in eggs from free-range hens than in all other environments. We did not expect egg fattyacid or carotenoid concentrations to vary between strains or by hen age.

Methods: Eggs from white egg layers (TA Tetra White) and brownegg layers (Hy-Line Brown), aged 44, 68, and 88 wk, were evaluatedfor hens reared in 1) conventional battery cages, 2) enrichable cagesystems, 3) enriched colony housing, 4) cage free (HB hens only),and 5) free-range rearing systems. Six randomly selected eggs fromeach set were homogenized for analysis. Lipids were extracted using amodified Folchmethod and fatty acidmethyl ester analysis for commonsaturated fatty acid (SFA), monounsaturated fatty acid (MUFA), andpolyunsaturated fatty acid (PUFA) species was performed by GC-flameionization detection. The extract was also analyzed for carotenoid andretinol content by HPLC-MS/MS.

Results: Eggs from enriched colony and free-range hens had higherMUFA and SFA content than eggs from conventional hens. The totalPUFA content of eggs from free-range hens was higher than that of eggsfrom conventional hens. However, arachidonic acid (20:4n–6) content


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was higher in eggs from enriched colony and free-range hens. Thetotal carotenoid content of eggs from free-range hens was almost twicethat of eggs from conventional hens. The retinol content was lowerin eggs from 88-wk hens than in eggs from 68-, but not 44-wk hens.Strain by age interaction effects were detected for egg content of α-linolenic acid (18:3n–3). Interaction effects between age, strain, andenvironment were detected for egg content of α- and β- carotene, luteinor zeaxanthin, and total carotenoids.

Conclusions: In our study, rearing condition, strain, and age wereassociated with differences in fatty acid, retinol, and carotenoid contentof eggs. With the exception of 20:4n–6, differences in fatty acid, retinol,and carotenoid are most likely not relevant to human nutrition at thecurrent level of egg consumption.

Funding SourcesThis research was supported by the North Carolina Layer Perfor-

mance andManagement Program, Raleigh, NC, theUSDA,ARS project3062-51,000-053-00D (to SKR and LH-M), and a grant from the EggNutrition Center, Park Ridge, IL (to SKR and LH-M).

Dietary PUFA, Postprandial Total Cholesterol, and TriglycerideConcentrations (P10-104)

Susan K Raatz, Vanessa Thyne, LuAnn Johnson, and MatthewPicklo

USDA

Objectives: The objective of this work was to determine the impactof the consumption of different dietary fatty acids on postprandial (PP)lipemia. We hypothesized that intake of unsaturated fatty acids resultsin lower triglyceride (TG) and total cholesterol (TC) concentrations inthe PP state compared with saturated fatty acid (SFA).

Methods: With preliminary data from an ongoing study, weexamined PP lipid responses over 4 h in volunteer participants. The trialwas a crossover-designed, acute study in which participants consumedeach of 5 treatments [SFA, monounsaturated fatty acid (MUFA),polyunsaturated fatty acid (PUFA)-α-linolenic acid (ALA), PUFA-linoleic acid (LA), and PUFA-long-chain omega-3 (LCn3)] in randomorder with a 1-wk washout between treatments. Participants wereadmitted to the research center the evening before testing, where theyreceived a standardized dinner meal and were then allowed nothingby mouth until testing. A fat tolerance test, provided as a beveragecontaining 30 g of total fat and enriched in the fatty acids of interest,was provided after the time 0 blood draw. Concentrations of TC, LDLcholesterol, HDL cholesterol, and TG were determined at 0, 30, 60, 120,180, and 240 min. The area under the curve (AUC; expressed in mg ·dL–1 ·h–1), corrected for the baseline for each lipoprotein,was calculatedby the trapezoid rule.

Results:Overweight to obese men (n= 2) and women (n= 6), aged26.1± 2.3 y (mean± SEM)with a bodymass index of 30.9± 0.8 kg/m2,were selected for inclusion in this analysis from an ongoing study. Nodifferences were observed in the AUC for HDL cholesterol or LDLcholesterol. The TC AUC was significantly reduced with PUFA-LCn3compared with SFA (P = 0.02). The TG AUC was significant reducedwith PUFA-ALA, PUFA-LA, and PUFA-LCn–3 intake compared withSFA (P = 0.001, P = 0.05, and P = 0.01, respectively).

Conclusions: Our preliminary data provide evidence that PP TGand TC AUC are reduced after consuming test meals which contain

PUFA compared with SFA. Further investigations are needed todetermine day-long interaction of dietary fatty acids with postprandiallipemia.

Funding SourcesThis research was supported by the USDA, ARS project 3062-

51,000-053-00D.

The Metabolic Availability of Methionine in Cooked CanadianChickpeas in Healthy Adult Men as Shown by the Indicator AminoAcid Oxidation Technique (P10-105)

Mahroukh Rafii,1 Paul Pencharz,1 Ronald Ball,2 ChristopherTomlinson, 3 Rajavel Elango,4 and Glenda Courtney-Martin1

1The Hospital for Sick Children, Canada; 2University of Alberta,Canada; 3University of Toronto, Canada; and 4BC Children’s Hospital,Canada

Objectives: The objective of this study was to determine the proteinquality of Canadian chickpeas by determining themetabolic availability(MA) of methionine. In addition, we aimed to assess the effect ofcomplementation of the chickpeas with rice on the MA of methioninein a mixed-meal format.

Methods: To determine the MA of methionine, 4 amounts ofmethionine were studied: 0.5, 1, 2, and 3 mg · kg–1 · d–1 (= 11.1, 22.2,44.4, and 66.7% of the mean methionine requirement of 4.5 mg · kg–1· d–1) either as L-methionine or from chickpeas in 6 young men ina repeated-measures design using the indicator amino acid oxidationmethod (IAAO), with L-[1-13C]phenylalanine as the indicator. Eachsubject received 4 intakes as L-methionine and 3 levels of intake fromchickpeas. The MA of methionine was estimated by comparing theIAAO response to varying intakes of methionine in cooked Canadianchickpeas compared with the IAAO response to L-methionine intakesin the reference protein (crystalline amino acid mixture patterned afteregg protein) using the slope ratio method.

Results: Preliminary data from 4 of the 6 subjects showed an MA ofmethionine from Canadian cooked chickpeas of 68%.

Conclusions:Our data suggest that, in addition to being limiting inmethionine, diets that provide chickpeas as the protein source need tobe corrected for bioavailability ofmethionine in order to correctly assessprotein quality.

Funding SourcesSeeding Food Innovation Grant, George Weston Ltd.

Hepatic FGF21 Is Required for Dietary Protein Restriction–Mediated Fatty Acid Oxidation by Activated Brown Adipose Tissue(P10-106)

Justin S Reynolds, Humberto Treviño-Villarreal, MichaelMacArthur, Alexandra Yesian, Sarah Mitchell, and JamesMitchell

Harvard TH Chan School of Public Health, MA

Background: Cold-activated brown adipose tissue (BAT) has themetabolic flexibility to oxidize glucose and fatty acids viamitochondrialuncoupling protein 1 (UCP1). Dietary protein restriction (PR) alsoactivates BAT and increases energy expenditure, including glucosedisposal, but whether this applies to fatty acids is unknown.


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Objective: The aim of this study was to test the hypothesis thatPR-induced BAT activation induces fat burning and overall adiposityreduction via the PR-responsive hepatic transcription factor cAMP-responsive element-binding protein H (CREBH) and its target, thehormone FGF21.

Methods: Mice were given ad libitum access to a semipurifiedcomplete diet (18% protein, 22% fat, 60% carbohydrate), or a protein-free diet in which protein was replaced with isocaloric sucrose, for 7 d atroom temperature (22ºC) or at thermoneutrality (30ºC). InterscapularBAT was analyzed using quantitative reverse transcriptase-polymerasechain reaction and immunoblotting. BAT substrate utilization wasinvestigated by measuring BAT [3H]palmitate or [14C]2-deoxyglucoseuptake. The role of FGF21 was explored using adenoviral short-hairpinRNA, and the roles of CD36 and CREBH were analyzed using globalknockout mice. Adiposity was measured with EchoMRI.

Results:PR led to a significant increase in the BAT activationmarkerUCP1, but not the traditional browning markers PRDM16, PGC1A,or CIDEA, at both room temperature and thermoneutrality, whereas

FIGURE 1 The roles of CD36, FGF21, and CREBH in temperature-independent PR-induced BAT activation. Fold change in BAT mRNAlevels (a,d), albumin-conjugated [3H]palmitate uptake in multiple tissues (b) or BAT (c), serum FGF21 (e), or change in body adiposity (f),following 7 days on the indicated diets, in mice at the indicated ambient temperature (a-b) or of the indicated genotypes (d-f) (n=4-10 pergroup). All data are expressed as mean ± SEM. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, using a two-way ANOVA with a Tukey’spost-hoc test.

type II iodothyronine deiodinase (DIO2) increased significantly atroom temperature alone. PR also significantly increased BAT [14C]2-deoxglucose and [3H]palmitate uptake. In mice lacking the fatty acidtransporter CD36, PR-mediated activation of UCP1 and DIO2 in BATwas maintained; however, the increase in [3H]palmitate uptake wassignificantly impaired. FGF21 knockdown significantly reduced PR-mediatedBATUCP1 induction and [3H]palmitate uptake. PR-mediatedhepatic FGF21 inductionwas also dampened inCREBHknockoutmice,concomitant with reduced UCP1 expression and [3H]palmitate uptakein BAT, and failure to reduce adiposity.

Conclusions:Activation of BAT by PR conferred metabolic flexibil-ity to oxidize glucose and fatty acids, but differed from cold-inducedBAT activation in gene expression and temperature independence. PR-mediated BAT activation, fat uptake, and reduction in adiposity allrequired FGF21, a liver-derived hormone controlled in part by CREBH.

Funding SourcesResearch was supported by NIH (R01DK090629, R01AG036712)

and Charoen Pokphand Group.


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SerumConcentrations ofObestatin andGhrelin inChildrenwithObesity and of Normal Weight (P10-107)

Enrique Romero-Velarde,1 Claudia Garcia Gonzalez,2 CarmenM Gurrola-Diaz,3 Martha P Sanchez Muñoz,2 Liuba M Aguirre-Salas,2 and Irma Soto-Luna3

1Universidad de Guadalajara, Mexico; 2Hospital Civil de Guadala-jara Dr. Juan I. Menchaca, Mexico; and 3CUCS, Universidad deGuadalajara, Mexico

Objective: The aim of this study was to demonstrate that the serumconcentrations of ghrelin and obestatin in children with obesity differfrom the concentrations of those with normal weight.

Methods: In a cross-sectional study, 42 children with obesity and 55with normal weight were included, with the consent of their parents,in the outpatient clinic at the Hospital Civil de Guadalajara Dr. Juan I.Menchaca. Subjects, aged 6–19 y, with exogenous obesity [body massindex (BMI) >2 SD] or normal weight (BMI between –1 and +1SD) were included. Non-probabilistic sampling was performed amongchildren who attended the outpatient clinic andmet the inclusion crite-ria. An interview was conducted with the parents and anthropometricmeasurements were performed. The next day, a fasting blood samplewas taken for determination of the hormones ghrelin and obestatin, andthemetabolic profile. Descriptive statistics were performed and ghrelin,obestatin, andmetabolic profile concentrationswere compared betweengroups of children with normal weight and obesity.

Results: The mean age was higher in children with normal weightthan in the obese group (11.6 vs. 9.8 y;P< 0.05). Ghrelin concentrationswere significantly lower in children with obesity (P < 0.05), whereasobestatin concentrations were lower in the normal weight group,but without statistical significance (P = 0.09). We did not find anydifferences according to sex. Concentrations of insulin, triglycerides,LDL, and VLDL were higher in the group of children with obesity(P < 0.05), whereas HDL was higher in the normal-weight children(P < 0.001).

Conclusions: The lower concentration of ghrelin in obese subjectsis apparently associated with negative feedback mechanisms of appetitecontrol. In the case of obestatin, more data are required with a greaternumber of subjects to confirm possible alterations in the concentrationand behavior in overweight or obese subjects. Our results suggest aprobable process of adaptation to weight gain, since ghrelin stimulatesweight gain and obestatin antagonizes the actions of ghrelin.

Funding SourcesConsejo Nacional de Ciencia y Tecnología and Universidad de

Guadalajara.

Sustained Effect Induced by Resistant Starch on the Regulationof the Expression of Genes Related to Digestion or Absorption ofCarbohydrates in the Small Intestine (P10-108)

Aratza Mireya Rosas Perez, Kazue Honma, and Toshinao Goda

University of Shizuoka, Japan

Background: Starch is one of the most abundant components infood products, and is the main source of digestible carbohydrates.Previous studies have demonstrated that feeding rats with dietaryresistant starch (RS) leads to the regulation of the expression of genesrelated to the digestion and absorption of starch.

Objective: The aim of this study was to evaluate whether a dietcontaining RS could lead to epigenetic control by suppression of genesrelated to the digestion and absorption of carbohydrates in the smallintestine, and to determine if this effect persists for several days.

Methods: Sprague-Dawley rats (n = 30) were fed with RS or acontrol diet for 14 d, then were fed a diet without RS for 4 d to assesswhether there were any sustained effects. We measured the enzymaticactivity of maltase-glucoamylase (MGAM) and sucrase-isomaltase (SI),and the expression of the corresponding genes and the Na+ glucosecotransporter (Sglt1), in different parts of the small intestine (upperand lower jejunum and ileum) to evaluate the distribution of thecarbohydrate digestive capability in rats.

Results: The results showed that there were significant decreases inthe activities of maltase, sucrase, and isomaltase in the upper jejunumafter 14 d with the RS treatment compared with the control, andthese correlated with reductions in gene expression of Mgam and Si,respectively. We also found that the gene expressions of Sglt1 and thehepatocyte nuclear factor 1a (Hnf1a) were downregulated by the RSfeeding, although the expression levels recovered during the following4 d. By contrast, the activities of maltase, sucrase, and isomaltase wereenhanced significantly in the lower jejunum after RS feeding, and theenhanced effect was retained for 2 d without further RS treatment.

Conclusion: These results suggest that the RS feeding can exert adownregulation of the genes related to the digestion and absorptionof starch in the upper jejunum, which gradually recovers during thefollowing 4 d.

Protein Intake and Risk of Falls in Older Adults: ProspectiveAnalysis and Meta-analysis of the Literature (P10-109)

Helena Sandoval-Insausti,1,2 Raúl F Pérez-Tasigchana,1,2Esther López-García,1,2 José R Banegas,1,2 Fernando Rodríguez-Artalejo,1,2 and Pilar Guallar-Castillón1,2

1School of Medicine, Universidad Autónoma de Madrid-IdiPaz,Spain; and 2CIBERESP (CIBER of Epidemiology and Public Health),Madrid, Spain

Objectives: The association between protein intake and falls hasbeen scarcely studied, and in older adults could change according totheir nutritional status. The aims of this study were to assess thisassociation in a Spanish cohort and to perform a meta-analysis.

Methods:Weundertook a prospective cohort study of 2464men andwomen aged ≥60 y, recruited in 2008–10 and followed up throughout2012. At baseline, habitual protein intake was determined with avalidated dietary history. At the end of the follow-up, participantsreported the number of falls they had suffered in the preceding year.Participants were stratified according to their nutritional status by usingunintentional weight loss of ≥4.5 kg. Logistic regression or negativebinomial regression was used, as appropriate. A systematic review ofthe literature was performed and the results were combined using fixed-effect meta-analytic models.

Results: A total of 522 participants (21.2%) suffered ≥1 fall, and163 (6.6%) had lost weight unintentionally. A statistically significantinteraction was observed with unintentional weight loss when assessingthe association of protein intake and risk of falls (P-interaction= 0.004).No association was found among participants without weight loss.However, among those with unintentional weight loss, theORs and 95%


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CIs of falling for increasing tertiles of total protein intake were 1.00,0.68 (95% CI: 0.21, 2.23), and 0.23 (95% CI: 0.05, 1.08); P-trend = 0.01.Similar results were obtained for animal and vegetable proteins. In themeta-analysis, the pooled incidence rate ratio for a 1 SD increase intotal protein intake among thosewith unintentionalweight losswas 0.63(95%CI: 0.41, 0.98). The corresponding values for animal and vegetableprotein intake were 0.67 (95% CI: 0.46, 0.96) and 0.57 (95% CI: 0.39,0.83), respectively.

Conclusions:Older adults who have suffered a decline in nutritionalstatus might decrease their risk of falls with increased dietary proteinintake.

Funding SourcesThe data collection was funded by the following grants: FIS

PI16/1512, PI16/1460, PI16/609, PI17/1709 (State Secretary of R + Dand FEDER/FSE), the FRAILOMIC Initiative (EU FP7-HEALTH-2012—Proposal No. 305,483-2), the ATHLOS project (EU H2020—Project ID: 635,316), the SALAMANDER project (PCIN-2016-145),and the CIBERESP, Instituto de Salud Carlos III. Madrid, Spain.

Effect of a High-Protein, High-Fiber Beverage Preload on Ap-petite and Energy Intake in Overweight Men and Women: Role ofAge and Dietary Restraint (P10-110)

Mastaneh Sharafi,1 Nima Alamdari,1 Michael Wilson,1 HeatherJ Leidy,2 and Erin Glynn1

1Beachbody, LLC; and 2Purdue University, IN

Objective: The purpose of this study was to determine the acuteeffects of a high-protein, high-fiber (HP/HFb) beverage taken asa preload compared with an isocaloric lower-protein, lower-fiber(LP/LFb) placebo beverage on subjective appetite ratings and subse-quent energy intake at an ad libitum meal in healthy adults.

Methods:A total of 50 overweight or obesemen andwomen (n= 25men, 25 women; age: 30 ± 2 y; body mass index: 29.6 ± 0.3 kg/m2)received a 160-calorie HP/HFb beverage containing 17 g of proteinand 6 g of fiber on one occasion and an isocaloric LP/LFb placebobeverage containing 1 g of protein and 3 g on fiber on another occasionin a randomized, double-blind, crossover design. At 30 min followingconsumption of the beverage preload, an ad libitum pizza meal wasprovided to be consumed over a 30-min period.Visual analog scalewereused to assess subjective appetite ratings throughout the testing period.The revised restraint scale was used to classify participants as restrainedor unrestrained eaters.

Results:HP/HFb led to greater reductions in postprandial desire toeat and hunger compared with LP/LFb (both, P < 0.05), but did notsignificantly affect postprandial fullness or prospective food consump-tion. Subsequent meal energy intake tended to be lower after HP/HFbcompared with LP/LFb (P = 0.09). A subanalysis showed lower energyintake after HP/HFb in older participants (≥25 y) compared withLP/Lfb, which was not observed in the younger participants (<25 y).

Conclusions: Compared with the LP/LFb, the HP/HFb beveragepreload reduced hunger and desire to eat, and tended to reducesubsequent food intake. Dietary restraint and age appear to influencesubsequent energy intake, and should be taken into account whendesigning nutrition interventions for weight reduction and/or mainte-nance.

This trial was registered at clinicaltrials.gov as NCT02979717.

Funding SourcesThis study was funded by Beachbody, LLC.

Calcium Ion Enhances Inhibition of Branched-Chain α-KetoacidDehydrogenase Kinase by Thiamine Pyrophosphate (P10-111)

Yoshiharu Shimomura, Seisuke Noguchi, and Yasuyuki Kitaura

Nagoya University Graduate School of Bioagricultural Sciences,Japan

Abstract: The main catabolic pathways of branched-chain aminoacids (BCAAs) are located in the mitochondria. The first 2 steps of thepathway are common to the 3 BCAAs (leucine, isoleucine, and valine).The first step is transamination of the BCAAs to form branched-chainα-ketoacids (BCKAs), catalyzed by branched-chain aminotransferase,and the second step is the oxidative decarboxylation of the BCKAs toproduce corresponding CoA esters, catalyzed by the branched-chainα-ketoacid dehydrogenase (BCKDH) complex. Since the latter is acommitted step in the catabolism of BCAAs, the regulation of theBCKDH complex is very important in the homeostasis of the freeBCAA concentrations. The BCKDH complex is regulated by covalentmodification, and BCKDH kinase (BDK) is responsible for inactivationof the complex by phosphorylation. Thiamine pyrophosphate (TPP) isknown as the coenzyme of the complex as well as the BDK inhibitor.In the present study, we found that free calcium ion in the physiologicrange greatly increases the sensitivity of BDK to TPP: inhibition ofthe BDK by TPP with a half maximal inhibitory concentration of 14and 0.11 µM in the absence and presence of 1 µM free calcium ion,respectively. These findings suggest a novel mechanism for the calciumion-dependent regulation of BCKDH complex.

Funding SourcesThis work was supported in part by JSPS KAKENHI (No. 17H03817

from Grant-in-Aid for Scientific Research (B) (to YS).

The Impact of Mechanistic Target of Rapamycin Nutrient-Sensing Metabolic Pathway and the 5’-AMP-Activated ProteinKinase Activator Metformin on the Growth of Pancreatic CancerCells Lines in C57BL/6 Mice (P10-112)

Ghada A Soliman,1 Surendra Shukla,2 Asserewou Etekpo,3Venugopal Gunda,2 Sharalyn Steenson,2 NagsenGautam,2 YazenAlnouti,4 and Pankaj K Singh5

1CUNY, Graduate School of Public Health and Health Policy,Affiliate Faculty, Advanced Science Research Center, City Universityof New York; 2University of Nebraska Medical Center; 3NebraskaDepartment of Health; 4College of Pharmacy and 5Fred & PamelaBuffett Cancer Center, University of Nebraska Medical Center

Objective: The aim of the study was to investigate the impact ofinhibition of the mechanistic target of rapamycin (mTOR) complexesand synergism with the antidiabetic drug metformin on pancreatictumor growth and autophagy. The hypothesis tested is that inhibitors ofmTOR complexes will synergize with 5′ -AMP-activated protein kinaseactivators to augment tumor responsiveness to treatments. mTOR is anutrient-sensing pathway and is dysregulated in pancreatic cancer andtype II diabetes.

Methods: Cell lines derived from pancreatic tumors of the triplemutant KrasG12D; p53R172H; Pdx1-Cre (KPC) transgenic mice were


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implanted into the pancreas of C57BL/6 mice (n = 10/group). After4 wk of tumor growth, the mice were injected intraperitoneally withdaily doses of low or high concentrations of 1) Torin 2 [mechanistictarget of rapamycin complex 1 (mTORC1) + mTORC2 inhibitor] high(TH), 2) Torin 2 low (TL), 3)metformin low (ML), 4) a combination ofTorin 2 and metformin low (TLML), or 5) dimethylsulfoxide (control)for 14 d. At the end of the treatments, the mice were killed, tumorweight and volume were measured, and blood samples and organs wereharvested and flash-frozen for metabolomics analysis.

Results: The drug distribution of Torin 2 and metformin in tissuesand plasma was confirmed by LC-MS/MS. Metabolomics analysis ofthe control and treated plasma samples showed differential metaboliteprofiles. Phenylalanine was significantly elevated in the TLML groupcompared with the control (+426%, P = 0.0004), whereas uracilwas significantly lower (–38%, P = 0.009), which may suggest thatautophagy is altered. The data also revealed that TLML significantlydecreased pancreatic tumor volume (498+ 104mm3; 37%; P< 0.0004)compared with control (1326 + 134 mm3; 100%), ML (853 + 67mm3; 64%), TL (745 + 167 mm3; 54%), and TH (665 + 182 mm3;50%), according to ANOVA and post hoc tests. However, TH wastoxic to the animals (only 3 out of 10 mice survived). Additionally,TLML significantly decreased the tumor weight (0.66 + 0.08 g; 52%),compared with the control (1.28 + 0.19 gm; 100%) (P < 0.002).

Conclusions: The data indicate that the combination of mTOR dualinhibition by Torin 2 and metformin is associated with a significantreduction in pancreatic tumor size and weight, and is better toleratedin mice. The findings suggest that combinations of drugs provide bettertumor reduction and might be a better target for treatment than single-agent therapies.

Funding SourcesCancer Prevention and Control Pilot Grant.

A Descriptive Analysis of Protein Intake Relative to Lean BodyMass (P10-113)

Joseph R Stanzione, Joseph Boullata, Michael L Bruneau, andStella L Volpe

Drexel University, PA

Background: Adult requirements for daily protein intake relyon nitrogen balance data and are described in terms of total bodyweight, regardless of body composition. Given the variability in bodycomposition, and that the nitrogen balance reflects the body cell massor its clinical proxy, lean body mass (LBM), it may be appropriate toconsider LBM when analyzing adult protein needs.

Objectives:The aim of this study was to describe the average dietaryprotein intake, expressed as g/kg LBM, in a group of healthy adultathletes.

Methods: Data were obtained from a cross-sectional study in acohort of healthy masters athletes. Participants’ body weight and heightwere measured using a mechanical column scale with an attachedstadiometer. Body composition was measured using dual-energy X-rayabsorptiometry. Each participant also completed a 2005 Block’s FoodFrequency Questionnaire. Descriptive statistics were used to determinethe average energy intake, the percentage of energy from protein andthe average protein intake in g/kg LBM. An independent samples t testwas used to determine if differences existed in protein intake between

women and men. A one-sample t test was used to compare the averageprotein intake to the Recommended Dietary Allowance (RDA; 0.8 g/kgbody weight) for both women andmen. Alpha levels were set a priori atP < 0.05.

Results:A total of 132 masters athletes (70 women, 62 men; 39± 10y of age; body mass index: 24.6 ± 3.6 kg/m2) were included in ouranalysis. Average total energy intake was 1887.4± 784.5 kcal/d. Averagepercentage energy intake fromproteinwas 15.5± 2.6%.Average proteinintake was 1.42 ± 0.54 g/kg LBM for the group. A significant differencewas evident between women andmen (1.51± 0.56 and 1.32± 0.50 g/kgLBM, respectively; P = 0.043). Both women and men had significantlyhigher protein intakes compared with the RDA (P < 0.001).

Conclusions: We were able to identify the average protein intake(g/kg LBM) for a group of healthy adults. In addition, we identifieddifferences in protein intake between women and men. Our findingsmay contribute to evolving perspectives on how best to determine adultprotein requirements.


Evaluation of Glycemic Response and Breath Hydrogen ofIsomaltooligosaccharides (P10-114)

Maria L Stewart, Maria Stewart, Didem Icoz, and VishnupriyaGourineni

Ingredion Inc.

Background: Ingredients that deliver functional (sweetness andtexture) and nutritional benefits are of interest to food manufacturers.Some oligosaccharides which serve as alternate sweeteners fit intothis category, such as isomalto-oligosaccharides (IMOs). CommercialIMOs are typically a mixture of α-(1→6) and α-(1→4) linked glucoseoligomers, synthesized by enzymatic reaction from starches such ascorn or tapioca.

Objective: The aim of this study was to evaluate the digestibility andfermentability of IMOs in a healthy population.

Methods: Two randomized, double-blind, placebo-controlled,crossover human studies were conducted. In the pilot study(n = 10), a 2-h postprandial glycemic response to BIOLIGO IL5040oligosaccharide and BIOLIGO IL7010 oligosaccharide was evaluatedand compared with dextrose (control). The tested IMOs differed in thecomposition of their mono- and disaccharide sugars. In the secondstudy (n = 26), the glycemic and insulinemic response to BIOLIGOIL5040 oligosaccharide was measured as a primary outcome. Breathhydrogen over 24 h and gastrointestinal tolerance were measured assecondary outcomes.

Results: In both studies, the IMO syrup doses were matched for50 g of total carbohydrates and were consumed by mixing in water(8 oz).

Conclusions: The incremental area under the curve for both IMOswas similar to dextrose. The lack of a difference in glycemic responses tothe IMOs, with no impact on breath hydrogen and intestinal tolerance,demonstrates that IMOs are digestible and can be used to replace sugarsin product formulations.

Funding SourcesIngredion Incorporated.


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Selective Decrease in Pentaenoic Long- and Very Long–ChainFatty Acids in the Lipid Classes of Underdeveloped Testis in ObeseZucker Rats (P10-115)

Miyoung Suh, Jutika Datar, Carla G Taylor, and Peter Zahradka

University of Manitoba

Objective: A previous study has shown that a significant decreasein n–6 (ω-6) docosapentaenoic acid (n–6 DPA) in phosphatidylcholineand phosphatidylethanolamine is associated with testis underdevelop-ment. The aim of this study was to investigate whether the same phe-nomenon takes place in other lipid classes in cases of underdevelopedtestis.

Methods: Adult male obese (fa/fa) Zucker rats presenting bothunder- and normal-sized testis were used for this study. Testis withmore than 30% weight differences in a paired testes was consideredunderdeveloped. n–6 Long-chain fatty acids and their elongated anddesaturated products of very long-chain fatty acids (VLCFA; C >24)weremeasured in phospholipid (PL), triacylglycerides (TG), cholesterolester (CE) and free fatty acids (FFA) in testis.

Results: In comparison to normal-size testis, underdeveloped testesshowed a marked decrease in n–6 DPA in PL and FFA, and total n–6 pentaenoic VLCFA in PL, TG, and FFA. In contrast, the precursorof DPA, 20:4n–6, was significantly increased in PL, and C22:4n–6 andC28:4n–6 were increased in PL, CE, and FFA.

Conclusions: This study indicates that conversion of tetraenoic topentaenoic long-chain fatty acids and VLCFA in underdeveloped testeswas limited in underdeveloped testis PL, TG, and FFA, indicating acritical role of fatty acids in testis development. These findings implicatethe lipid involvement in male fertility.

Funding SourcesUniversity of Manitoba Research Grant Program and Natural

Sciences and Engineering Research Council (NSERC) of CanadaDiscovery Bridge Funding Program.

The ATP-Sensitive K+ Channel KATP Mediates Taste ReceptorType 1–Independent Caloric Sugar Taste Signaling (P10-116)

Sunil K Sukumaran,1 Karen Yee,1 Shusuke Iwata,2 TimGilbertson,3 Buford Nichols,4 Noriatsu Shigemura,2 YuzoNinomiya,2 and Robert F Margolskee1

1Monell Chemical Senses Center, PA; 2Kyushu University, Japan;3Utah State University; and 4Baylor College of Medicine, TX

Abstract: The heterodimeric G-protein coupled receptor formedby the taste receptor type 1 proteins T1R3 and T1R2 is the primarytransducer of sweet taste, and is capable of sensing all categories of sweettastants, i.e., caloric and noncaloric sweeteners and sweet proteins.However, T1R3 knockout mice show significant, albeit diminished,behavioral and taste nerve responses to caloric sugars but not toartificial sweeteners, indicating that additional pathway(s) that aretuned exclusively to caloric sugars may exist. We hypothesized thatglucosidases, glucose transporters, and the ATP-sensitive potassiumK+

channel (KATP), which together mediate caloric sugar sensing in tissuessuch as the intestine, pancreas, andhypothalamusmay also do so in tastecells. In agreement with this hypothesis, cell type–specific expression

of the α-glucosidases α-amylase, sucrase-isomaltase, and maltase-glucoamylase, sodium-glucose cotransporters, several facilitative glu-cose transporters, and subunits of the KATP channel were detected inT1R3-expressing taste cells in mice and humans. Functional studies inmice showed that inhibitors of α-glucosidases partially abrogated tastenerve responses to disaccharides such as sucrose and maltose whileleaving responses tomonosaccharides, noncaloric sweeteners,and othertaste qualities, such as umami, bitter, and sour, unaffected. We couldalso detect KATP-mediated potassium currents in mouse taste cells.Our studies show that sweet taste signaling is complex and raise thepossibility of developing novel types of noncaloric sweeteners targetingcomponents of the KATP-mediated sweet taste signaling pathway.

Funding SourcesThis study was funded by NIH-NIDCD grants RO1 DC014105 and

P30 DC011735 to RFM.

Sodium Butyrate Induces Autophagy in Colorectal Cancer Cellsthrough LKB1/AMPK Signaling (P10-117)

Suxia Sun

No affiliation

Background: Butyrate is produced by the fermentation of undi-gested dietary fibers and is a promising candidate for use in cancertreatment. Autophagy is an evolutionarily conserved physiologicalresponse that is critical for advanced cancer progression. However, themechanisms underlying sodium butyrate (NaB)–induced autophagy incolorectal cancer, the third most common cancer and third leadingcause of cancer death worldwide, are not yet completely understood.

Objective: The aim of this study was to elucidate the mechanismsunderlying NaB-induced autophagy in colorectal cancer.

Methods: The expressions of LC3-II protein and mRNA weredetected by Western blot assay and quantitative reverse transcriptase-polymerase chain reaction, respectively. Autolysosome formationwas observed by transmission electron microscopy. Adenosinemonophosphate–activated protein kinase (AMPK) and liver kinase B1(LKB1) were inhibited by chemical inhibitor or siRNAs and confirmedby Western blot assay.

Results: NaB dose-dependently increased the expressions of LC3in protein and mRNA levels. We also found that the formationof autolysosome, expression of phosphorylated LKB1, AMPK, andacetyl-CoA carboxylase were increased in response to NaB treatment.Treatment with Compound C, an inhibitor of AMPK, as well as geneticinhibition of AMPKand LKB1 by siRNAs, significantly attenuatedNaB-induced autophagy in CRC cells.

Conclusion: Collectively, we indicated that LKB1 and AMPK arecritical for NaB-mediated autophagy and may act as novel targets forcolorectal cancer therapy in the future.

Funding SourcesThis study was supported by the grants from the National Natural

Science Foundation of China (Nos. 81,773,429 and 81,202,204).

The Effect of Very Low–CalorieDiets onHepaticOutcomes (P10-118)

Vijaya Surampudi, Xinkai Zhou, Chi-Hong Tseng, and ZhaopingLi

UCLA


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Objective: Very low–calorie diets (VLCDs) are an effective treat-ment option for obesity. There is evidence for the safe use of VLCDs,but there remains limited evidence of their effect on specific diseases.The aim of the present study was to perform a retrospective analysisin overweight or obese patients with possible nonalcoholic fatty liverdisease (NAFLD) manifested as elevated transaminases in a medicallysupervisedweightmanagement programusing high-proteinVLCDandlow-calorie diets (LCDs).

Methods: Patients were enrolled in a self-paid, university-based,medically supervised outpatient weight loss program that used a VLCD(500–800 kcal) or an LCD (800–1200 kcal). Patients came in weeklyand comprehensivemetabolic panel, complete blood count, lipids, bodycomposition, and electrocardiogram were measured. Patients who hadelevated transaminases without any evidence of hepatitis at baselinewere compared with the control group, who had normal transaminases.

Results: A total of 4620 patients were enrolled in the program from1991 to 2016. Of these patients, 3117 (84% female; 16% male) hadnormal transaminases (normal group) and 1503 (51% female; 49%male) were found to have elevated transaminases (NAFLD group). Themean body weight at baseline was 224.12 ± 60.85 lbs (mean ± SD)for the normal group and 244.38 ± 63.86 lbs for the NALFD group.The percentage weight loss within 6 mo was 10.53% for the normalgroup and 11.36% for the suspected NAFLD group. There was noclinically significant difference in transaminase values from baselineto 6 mo in the normal group, with mean aspartate aminotransferase(AST) and alanine aminotransferase (ALT) concentrations of 18.98 ±5.26 and 18.98 ± 4.60 units/L, respectively, at baseline and 21.39 ±10.92 and 21.34 ± 10.20 units/L, respectively, at 6 mo. In contrast, theNAFLD group had a clinically and statistically significant difference intransaminase values from baseline to 6 mo, with respective mean ASTand ALT concentrations of 49.94 ± 26.46 and 33.77 ± 16.69 units/L atbaseline and 30.93 ± 20.53 and 25.79 ± 16.81 units/L at 6 mo.

Conclusion: Our data suggest that patients who entered the pro-gram with elevated transaminases were able to reduce them effectivelywith a VLCD or LCD over 6 mo via weight loss. The data supportsthat VLCDs and LCDs are safe and can be more widely used in theprevention and treatment of obese patients with possible fatty liverdisease.


Bright Line Eating: An Effective, Online Program for SustainedWeight Loss (P10-119)

Andrew K Thaw and Carley Thaw

Millsaps College

Objective: Very low–calorie diets (VLCDs) are an effective treat-ment option for obesity. There is evidence for the safe use of VLCDsbut there remains limited evidence on specific diseases. The presentstudy was a retrospective analysis in overweight or obese patientswith possible nonalcoholic fatty liver disease (NAFLD) manifested aselevated transaminases in a medically supervised weight managementprogram using high protein VLCD and low calorie diets (LCD).

Methods: Patients were enrolled in self-paid, university based,medically supervised outpatient weight loss program that utilize VLCD

(500–800 kcal) or LCD (800–1200 kcal). Patients came in weekly andCMP,CBC, lipids, body composition andEKGweremonitored. Patientswho had elevated transaminases without any evidence of hepatitisat baseline were compared with the control group who had normaltransaminases.

Results: A total of 4620 patients were enrolled in the program from1991 to 2016. Of these patients 3117 (84% female; 16% male) patientshad normal transaminases (normal group) and 1503 (51% female; 49%male) patients were found to have elevated transaminases (NAFLDgroup). The mean body weight at baseline was 224.12 lbs (sd 60.85)for the normal group and 244.38 lbs (sd 63.86) for the NALFD group.Percent weight loss within 6 mo was 10.53% for the normal groupand 11.36% for the suspected NAFLD group. There was no clinicallysignificant difference in transaminase values from baseline to 6 mo inthe normal group with a mean aspartate aminotransferase (AST) andalanine aminotransferase (ALT) respectively of 18.98 unit/L (sd 5.26)and 18.98 unit/L (sd 4.60) at baseline; 21.39 unit/L (sd 10.92) and 21.34unit/L (sd 10.20) respectively at 6 mo. In contrast, the NAFLD grouphad a clinically and statistically significant difference in transaminasevalues from baseline to 6 mo with mean AST and ALT respectively of49.94 unit/L (sd 26.46) and 33.77 unit/L (sd 16.69) at baseline and ASTand ALT respectively of 30.93 unit/L (sd 20.53) and 25.79 unit/L (sd16.81) at 6 mo.

Conclusion: Our data suggests that patients who entered theprogram with elevated transaminases were able to improve theirtransaminases effectively with VLCD or LCD over 6 mo via weight loss.The data supports that VLCD and LCD are safe and can be more widelyused in the prevention and treatment of obese patients with possiblefatty liver disease.

Funding SourcesN/A.

Potential Effect of Perilla Oil on High Fat Diet–Induced ColonicInflammation in C57BL/6J Mice (P10-120)

Shalom Sara Thomas,1 Youn-Soo Cha,1 and Kyung-Ah Kim2

1Chonbuk National University, South Korea; and 2ChungnamNational University, South Korea

Background: The leaves and seed oil of Perilla frutescens are usedwidely for edible purposes in East Asian countries such as Korea, Japan,and China. Especially in Korea, perilla oil (PO) is used for cooking andseasoning purposes, and the leaves are used tomake pickles. PO is a richsource of polyunsaturated fatty acids (PUFAs). Consumption of a high-fat diet induces obesity and consequent metabolic syndrome associatedwith impaired lipid metabolism and chronic low-grade inflammation.Colitis is an inflammatory disorder that mainly affects the colon part ofthe digestive tract, and is characterized by pathologic mucosal damageand ulceration. Dietary n–3 PUFAs are reported to have an amelioratingeffect in a 2,4,6-trinitrobenzene sulfonic acid–induced colitis mousemodel.

Objective: The aim of the present study was to investigate the effectof α-linolenic acid–rich PO on a high-fat diet–inducedmouse model ofcolitis.

Methods: C57BL/6J mice, 8 wk old, were divided into 3 groups.These were fed a normal diet (ND, 10% kcal fat), a high-fat diet (HD,


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60% kcal fat), or a high-fat diet supplemented with PO (HPO; 8% oil ina 60% high-fat diet) for a period of 16 wk. Body weight was measuredonce a week and feed intake was measured twice a week. Serum wasanalyzed for lipid content, and inflammatory biomarkers such as tumornecrosis factor α, interleukin (IL)-1β and IL-6 were measured in serumand colon using commercially available kits.

Results: There was a significant reduction in the body weight ofthe HPO group compared with the HD group. Significantly reducedconcentrations of proinflammatory cytokines were also observed inboth serum and colon in the HPO group compared with the HD group.

Conclusions: PO treatment prevented weight gain and improvedproinflammatory biomarkers in the high-fat diet–induced colitis mousemodel, suggesting that PO has potential benefits against a number oflifestyle diseases, including obesity and other metabolic disorders.

Funding SourcesThis work was supported by a National Research Foundation of

Korea (NRF) grant funded by the Korea Government (Ministry ofEducation) (2017R1D1A1B03029418).

Regulation of Amino Acid Catabolism by Hepatocyte NuclearFactor 4α and Peroxisome Proliferator-Activated Receptor α PPARα

or Retinoid X Receptor α in the Liver (P10-121)

Sandra Tobon,1 Nimbe Torres,2 and Armando Tovar2

1UNAM-INCMNSZ, Mexico; and 2INCMNSZ, Mexico

Background: There is evidence that peroxisome proliferator–activated receptor α (PPARα) can function as a physiologic switchto preserve body proteins during a shift in the type of energy fuelby suppressing amino acid oxidation. However, it has been suggestedthat PPARα mediates this effect by interacting with other transcriptionfactors.

Objective: The aim of this study was to assess the potentialinteraction betweenPPARα, hepatocyte nuclear factor 4α (HNF4α) andretinoid X receptor α (RXRα) as a regulator of amino acid catabolism.

Methods: Co-immunoprecipitation assays were used to demon-strate the interaction ofHNF4α andRXRα. The effect of overexpressionof PPARα or RXRα on the expression of HNF4α was analyzed byWestern blot analysis. Additionally, the effect of HNF4α and PPARα

or RXRα on the activity of the serine dehydratase (sds) promoter, anamino acid–catabolizing enzyme, was tested using the luciferase genereporter assay.

Results: We found that PPARα interacts with HNF4α throughRXRα, and that overexpression of PPARα stabilizes the expression ofHNF4α, whereas when RXRα was overexpressed there was a decreasein the stability of HNF4α. These interactions were then tested on thesds promoter coupled to the luciferase reporter gene, and we found thatPPARα or RXRα did not have a direct effect on the luciferase promoter,unlike HNF4α. When HNF4α and PPARα were cotransfected, therewas no repression on sds promoter activation; however, when HNF4α,PPARα and RXRα were cotransfected, there was significant repressionof the sds promoter.

Conclusions:Our results indicate that the regulation of the sds genepromoter was positively activated by HNF4α, but its effect was highlyrepressed by the overexpression of the PPARα or RXRα heterodimer.This evidence indicates that ligands of PPARα, such a fatty acids, can

actively repress amino acid oxidation by stimulating the formation ofthe PPARα or RXRα complex and increasing HNF4α degradation.

Funding SourcesCONACYT.

Nitrogen Balance in Children and YoungAdults with Epidermol-ysis Bullosa (P10-122)

Camille Cristine Gomes Togo,1 Ana Paula Zidorio,1 PatriciaBotelho,2 Kenia de Carvalho,1 and Eliane Dutra1

1Postgraduate Program in Human Nutrition and 2Department ofNutrition, University of Brasília, Brazil

Objective: The aim of this study was to evaluate the proteincatabolism by nitrogen balance (NB) in children and young adults withepidermolysis bullosa (EB).

Methods:A case series report study was developed with 11 childrenand young adults with EBwhowere attending theUniversityHospital ofBrasília. Protein catabolism was evaluated, measuring the NB. This wascalculated using 2 equations: the classic equation for healthy individualsand the equation for burned individuals, as in burn patients, subjectswith EB may have increased catabolism owing to the presence ofwounds. The burn equation was then modified by using the bodysurface area (BSA) of noninfected and infected wounds in place ofthe BSA of second- and third-degree burns, respectively, to attempt toaddress the effects of wounds and infection seen in EB, which havepreviously been compared with burns patients. The protein intake wasevaluated by 24-h recall and the nitrogen excretion rate was evaluatedby 24-h urine analysis.

Results: The mean age was 16.36 ± 8.33 y, 54.54% (n = 6) werefemale, 27.27% (n = 3) clinically presented EB simplex, and 72.72%(n = 8) presented recessive dystrophic EB. Among the 3 participantswith EB simplex, 2 had only noninfectedwounds and the other had bothtypes of wounds. Among the participants with recessive dystrophic EB,5 had only noninfected wounds, 1 had only infected wounds, and theother 2 had both types of wounds. In relation to protein catabolism,10 individuals were classified with anabolism and only 1 was classifiedwith mild catabolism by the classic equation. By the adapted equationfor EB, 5 individuals were classified in anabolism, of which 3 had EBsimplex. In addition, 3 individuals were classified with mild catabolismand 3 individuals with moderate catabolism. All these individuals hadrecessive dystrophic EB. None of the patients were known to have anyrenal dysfunction.

Conclusions: The adapted equation seems to be the most suitableto evaluate the degree of protein catabolism in patients with EB. Usingthis equation, all the individuals whowere assessed to have some degreeof catabolism had recessive dystrophic EB, whereas all subjects with EBsimplex were categorized with anabolism. Thus, in this small selectionof patients, negative NB was observed, especially among subjects withrecessive dystrophic EB.

Funding SourcesThe Fundação de Apoio à Pesquisa do Distrito Federal (FAPDF)

supported this research presentation at the congress in Boston. CamilleTogo and Ana Paula Zidorio have a scholarship from Coordenação deAperfeiçoamento de Pessoal de Nível Superior (CAPES)—Brasil.


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Deuterium Compared with 15N-Labeling in Determining theDigestive and Metabolic Fate of Milk Proteins (P10-123)

Dalila Azzout-Marniche,1,2,3 Juliane Calvez,3 NadezdaKhodorova,3 Joseph Tessier,3 Romain Kapel,4 and ClaireGaudichon1

1AgroParisTech; 2UMR PNCA; 3INRA; and 4CNRS

Background: Bioavailability of indispensable amino acids (IAAs) isa determinant factor of protein quality. The intrinsic labeling of dietaryproteins with 15N is usually used to determine the digestive fate of IAAs,but cannot be applied to all protein sources. Deuterated water is analternative, but its accuracy has not been assessed.

Objective:We aimed to compare the performance of 15N and 2H fordetermination of protein digestibility and AA bioavailability.

Methods: Milk was labeled from 4 goats receiving orally 5 g of15N ammonium sulfate (99%). Either 80 or 160 ml of 2H2O (98%)was given in drinking water for 1 or 3 d, and milk was collected.Proteins were purified from the milk and their 15N and 2H enrichmentsdetermined. Protein digestibility and AA bioavailability were assessedin 6 rats receiving a single test meal. Dietary proteins and AAswere quantified in gastrointestinal segments by elemental analysis-isotopic ratio mass spectrometry (IRMS) and GC-combustion-IRMSrespectively.

FIGURE 1

Results: 15N enrichment was 1.00% ± 0.07% in milk proteins, andthe AAs showed a uniform 15N enrichment of 0.94% ± 0.11% onaverage. The best 2H enrichment in milk proteins (0.18%) was obtainedwith the highest dose of 2H2O administered over 3 d. In contrast to15N, 2H enrichment in milk IAAs was not uniform, varying from 0.03%for phenylalanine to 0.11% for proline. In vivo, no 2H enrichmentwas found in the colon, contrary to 15N, leading to a slightly higherorocecal protein digestibility when assessed with 2H (98.0% ± 0.1% for2H vs. 97.6% ± 0.1% for 15N, P = 0.0004). However, orocecal proteindigestibility was similar with both tracers. With 15N labeling, the meanAA bioavailability of milk was estimated to be 98.0% ± 0.3%, varyingfrom 95.8% ± 1.6% (valine) to 99.1% ± 0.3% (methionine and lysine).The mean AA bioavailability determined with 2H was significantlylower than that determined with 15N (96.0% ± 0.8%, P = 0.0006), andvaried from 80.3% ± 2.4% (threonine) to 98.7% ± 0.4% (methionine).

Conclusions: Deuterium labeling of dietary protein appears asconvenient as 15N to follow the digestive fate of proteins. However,AA bioavailability seems to be underestimated with 2H compared with15N. Differences in cecum and colon bacterial metabolism as well astransamination of hydrogen exchange mechanisms may partly explainthese discrepancies.

Funding SourcesAgroParisTech, INRA, Dutch Dairy Association.


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TABLE 1 Bioavailability of indivisual AA assessed with 15N or 2H

Differentiation of Mediterranean and Western Dietary PatternsinNonhumanPrimates by theUse of BroadMetabolomics (P10-124)

Janet A Tooze, Jaime Speiser, Mara Z Vitolins, Susan Appt,Katherine L Cook, Thomas Register, and Carol Shively

Wake Forest School of Medicine, NC

Background: A Mediterranean diet pattern has been associatedwith health benefits when compared with a Western diet pattern.However, quantifying dietary patterns using self-report of diet may beproblematic, owing to systematic and random errors. Biomarker studiesof dietary intake, including metabolomics, may provide a means tocharacterize dietary patterns, but validation studies are needed.

Objective: We sought to identify broad metabolomic markers todistinguish Mediterranean and Western dietary patterns in a feedingstudy of nonhuman primates.

Methods: Urine and blood were collected from 37 female nonhu-man primates who were randomized to a Mediterranean or Westerndiet; the diet had been consumed for 2 y at the time of study. Bothdiets contained complete vitamin and mineral mixes for primates,ensuring adequate nutrition and comparable compositions of totalprotein, carbohydrates, and fat. Metabolites were measured using ultra-performance liquid chromatography and a high-resolution mass spec-trometer. Random forest analysis was used to identify the metabolitesthat most strongly differentiated the 2 diet groups. Following logtransformation, concentrations of specific metabolites were comparedbetween groups using Welch’s t test.

Results:The random forest classification analysis of both the plasmaand urine metabolic profiles was 100% accurate for differentiating the2 groups. For both plasma and urine analysis, metabolites that distin-guished the Western diet group from the Mediterranean diet groupincluded those associated with lipid intake andmetabolism, amino acidmetabolism, diet-derived phenolic metabolites, and micronutrients.

Conclusion: This long-term well-controlled feeding study demon-strated thatmetabolomics is a promisingmethod for identifying dietarypattern signatures, and warrants further evaluation.

Funding SourcesWake Forest Baptist Comprehensive Cancer Center andNCI Cancer

Center Support Grant P30 CA012197.

The Influences of Starch Fine Structure on the Digestion Mecha-nism of Human Pancreatic α-Amylase (P10-125)

Min-Hui Tsai,1 Chen Chen,1 and Amy Hui-Mei Lin1,2

1University of Idaho; and 2Washington State University

Background: Starch is the major glycemic carbohydrate providingdietary glucose to human. However, overconsuming rapidly digestiblestarch is associated with some health concerns, such as type 2 diabetesand obesity. We hypothesize that the structure of internal amylopectin(iAmp) determines the hydrolysis mechanisms of human pancreaticα-amylase (HPA), which is not well studied yet, and influences theglucogenesis.

Objective: The aim of this study was to investigate the digestionefficiency and mechanism of HPA on starches in vitro to betterunderstand the role of HPA in the small intestine and to provide anutrition strategy for modulating the postprandial glycemic response.

Methods: To characterize the structure of iAmp, starch washydrolyzed by α- and β-amylase, fractioned by size-exclusion columns,and analyzed by anion-exchange chromatography. To examine theα-amylase hydrolysis mechanism, starches were gelatinized and hy-drolyzed in vitro by HPA. The increase of reducing power and thestructure of the hydrolysates were examined. In vitro glucogenesiswas examined by applying mucosal α-glucosidase isolated from ratintestine. The results were evaluated statistically by one-way ANOVA.

Results: Waxy rice, with the shortest chain length of iAmp, hadthe highest production rate of reducing power. Waxy cassava and waxypotato, with long chain lengths of iAmp, had lower rates comparedwith waxy rice. The data on the hydrolysate structure showed that HPAwas capable of hydrolyzing the iAmp. A long iAmp chain (i.e., waxypotato and cassava) was cleaved by HPAmultiple times before the HPAattacked another amylopectin chain; a short iAmp chain (i.e., waxy rice)


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was cleaved only once, and the HPA continually attacked other chainsbecause the chain length was too short to allow formultiple cleaving. Asa result, the hydrolysis ofwaxy rice starch had generated a larger numberof small molecules, which were the favored substrates for mucosal α-glucosidase to generate glucose.

Conclusion: The structure of iAmp plays an important role indetermining HPA digestion. Selecting starch with proper amylopectinis a new strategy for modulating the postprandial glycemic response.

Assessment of the Protein Quality of IndianWheat byMeasuringthe Metabolic Availability of Lysine (P10-126)

Zujaja Tul-Noor,1 Paul Pencharz,2 Ronald Ball,3 Rajavel Elango,4and Glenda Courtney-Martin2

1University of Toronto, Canada; 2The Hospital for Sick Children,Canada; 3University of Alberta, Canada; and 4BC Children’s Hospital,Canada

Objectives: The aims of this study were 1) to assess the proteinquality of Indian wheat by studying the metabolic availability (MA)of the most limiting amino acid, lysine, and 2) to assess the effectof protein complementation by providing a mixed meal of wheat andlentils.

Methods: MA was studied with a repeated-measures design usingthe indicator amino acid oxidation (IAAO) method in 5 young menwith L-[1-13C]phenylalanine as the indicator amino acid at 4 levels oflysine: 5, 8, 12, and 15 mg · kg–1 · d–1 (i.e, 13.6, 21.7, 32.5 and 40.7%of the mean lysine requirement of 36.9 mg · kg–1 · d–1, respectively).Each subject participated in 8 experiments: 4 lysine intakes as L-lysinefrom crystalline amino acid, 3 intakes of lysine from wheat preparedas a bread, and 1 experiment for dietary complementation with lentils.Based on the principles of the slope-ratio assay, the MA of lysinewas calculated by comparing the IAAO response to varying intakes ofprotein-bound lysine in baked wheat bread with the IAAO responseto free crystalline L-lysine in the reference protein (patterned after eggprotein).

Results: Preliminary data from 4 of the 5 subjects show an MA oflysine from baked Indian wheat bread of 90%.

Conclusions:Despite being of limiting concentration inwheat,mostof the lysine present is available for protein synthesis when wheat isbaked to a golden color bread.

Funding SourcesSupported by the Canadian Institute of Health Research (FRN

148,864) and Graduate Restracomp Scholarships by the Hospital forSick Children Research Institute.

A Fads2 Inhibitor in Combination with Dietary n–3 Polyunsatu-rated Fatty Acids Alter Metabolic Parameters, but Not LymphocyteProliferation (P10-127)

Marie van der Merwe, Melissa Puppa, and Sunita Sharma

University of Memphis, TN

Objective: The objective of this study was to determine the effectof inhibition of Fads2, the rate-limiting enzyme in polyunsaturatedfatty acid (PUFA) metabolism, in combination with increased dietaryn–3 (ω-3) PUFAs onmetabolic and immune parameters, when given tohigh-fat diet–fed mice.

Methods: C57BL/6 male mice (n = 20), 7 wk old, were placedon a 45% high-fat diet (HF-n6, with an n–6:n–3 PUFA ratio of13:1) for 6 wk. Half of the animals (n = 10) were then switchedto a 45% high-fat diet containing large amounts of n–3 fatty acids(HF-n3, with an n–6:n–3 PUFA ratio of 1:3), predominantly in theform of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA).Concurrently, a subgroup receiving either diet (n = 5) was given dailyoral doses of a Fads2 inhibitor at a dose of 100 mg · kg–1 · d–1 for2 wk. This inhibitor will reduce synthesis of all Fads2 products,including arachidonic acid (AA), EPA, and DHA. Food consumptionand mouse weight were monitored throughout the experiment. Fastingglucose, glucose tolerance, and body composition were determinedpre- and postintervention. Mice were killed by CO2 inhalation andtheir organs harvested for characterization. Splenocytes were isolatedand stimulated with either lipopolysaccharide or concanavalin A todetermine their in vitro proliferation capacity.

Results:Consistent with the diet-induce obesitymodel, mice fed theHF-n6 diet gained significantly more weight compared with the chowdiet group (P < 0.01), but inhibition of Fads2, with or without the HF-n3 diet, reduced weight gain (P < 0.01). The loss of weight was a resultof fat loss (P < 0.01) as there was no change in lean mass. Interestingly,animals fed the HF-n3 diet in the absence of the Fads2 inhibitor hada slight increase in lean mass (P = 0.06). This was consistent with anincrease in the size of the gastrocnemius muscle in mice fed the HF-n3 diet (P = 0.06). Although no difference in glucose tolerance wasobserved, fasting glucose tended to be lower in the presence of the Fads2inhibitor, irrespective of diet. To determine the effect of inhibition ofFads2 on immunity, splenic B- and T-cell proliferation indices weredetermined. No differences were observed in any of the proliferationindices measured.

Conclusion: Inhibition of Fads2, with or without increased n–3PUFAs, can alter body composition but does not affect the proliferationcapacity of splenic lymphocytes.

Funding SourcesFunding was from aUniversity ofMemphis School of Health Studies

Faculty grant.

Comparison of the Effects of Soluble Corn Fiber and Fructo-oligosaccharide onMetabolism, Inflammation, andGutMicrobiomeof High-Fat Diet–Fed Mice (P10-128)

Matthias Van Hul,1 Hubert Plovier,1 Kavita Karnik,2 KirstieCanene-Adams,2 Mervyn De Souza,2 Amandine Everard,1 andPatrice Cani1

1Université catholique de Louvain, Belgium; and 2Tate & Lyle

Background: The health benefits of dietary fibers are now well rec-ognized. These include direct effects on gut functions and digestion, butalso indirect effects on blood glucose control, cardiovascular functions,and host metabolism. Yet the average intake remains far below rec-ommendations. Fiber supplementation therefore represents a realisticand efficient dietary intervention. Several products are already commer-cially available. Since different fibers may impact host health via differ-ent mechanisms, it remains critical to evaluate their physiologic effects.

Objective: The aim of this study was to assess the effects of 2types of fibers: soluble corn fiber (SCF; Promitor, Tate & Lyle) and


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fructo-oligosaccharide (FOS; Orafti P95, Beneo) in a mouse model ofdiet-induced obesity.

Methods: C57BL/6J male mice (n = 15/group) were fed a control(CT) or high-fat diet (HFD) supplemented without or with SCF or FOS(10% supplemented in water) for 8 wk. Body weight and composition(lean and fat mass) were recorded weekly. Inflammatory markers wereassessed by quantitative polymerase chain reaction and gut microbialcomposition by 16S rRNA sequencing.

Results: SCF and FOS decreased body weight gain and fat masssimilarly without affecting caloric intake. However, energy excretion(measured by bomb calorimetry) was increased by SCF but not byFOS, suggesting different mechanisms of action. For both fibers, therewas a reduction in the inflammation of liver (reduced monocytechemoattractant protein-1 expression) and adipose tissues (reducedlevels of lipopolysaccharide-binding protein, monocyte chemotacticprotein-1, CD11c), as well as an improvement in the glucose tolerance.Interestingly, the overall improved metabolic profile was associatedwith a significant impact on the gut microbial composition, whichshowed similarities between fibers (e.g., restoration of Pseudomonas,Dehalobacterium, Sutterella, and butyrate-producing Allobaculum lev-els; reduction of several HFD-induced bacteria and pathogens Bacillusand Desulfovibrio), but also differences (e.g., differential changes inBlautia, Coprococcus, and Parabacteroides levels, as well as that ofBifidobacterium prevotella and Lactococcus).

Conclusion: Although the dietary fibers SCF and FOS have similarphysiologic outcomes on mice fed an HFD, their mechanisms of actionappear to be different.

Funding SourcesThe research is supported by Tate & Lyle Ingredients Americas LLC.

Does Shape of Visceral Adipose Tissue InfluenceCardiometabolic Risk? Observations in a Multiethnic Cohort(P10-129)

Rosa C Villegas-Valle,1 Unhee Lim,2 Thomas Ernst,3 Bo Fan,4Steven D Buchthal,2 Bennett Ng,5 Amir PashaMahmoudzadeh,4Gerardo Alvarez-Hernandez,6 Carmen Candia-Plata,6 RolandoGiovanni Diaz-Zavala,6 Lynne Wilkens,2 Mauro E Valencia,6Loïc Le Marchand,2 and John Shepherd2

1University of Sonora, Graduate Program on Chemical Biologicaland Health Sciences, Mexico; 2University of Hawaii Cancer Cen-ter; 3University of Hawaii; 4University of California San Francisco;

5University of California Berkeley and University of California SanFrancisco Graduate Program in Bioengineering; and 6University ofSonora, Mexico

Objectives:Wehypothesized that the shape of visceral adipose tissue(VAT) is associated with metabolic syndrome (MetSx) in adults fromdifferent ethnic groups and aimed to determine this association.

Methods: Data from 1096 participants in a cross-sectional studywere analyzed to associate the abdominal shape, texture, and variationin appearance with the presence of MetSx. Magnetic resonance imagesof the abdomen at the L4-L5 intervertebral space were annotated using55 landmark points to define the abdominal tissues (VAT, subcutaneousfat, muscle) using statistical appearance modeling (SAM) as describedby Cootes. The SAM generated principal components (PCs) that reflectdominant modes of variation in the abdominal structures. In thisanalysis, only VAT PCs were included as independent variables inmultiple logistic regression models to predict the presence of MetSx.Demographic, body size, and adiposity variables (including VAT cross-sectional area as a measure of VAT quantity) were included in themodels to test if the PCs provide independent metabolic risk predictionbeyond VAT amount.

Results: A preliminary analysis of VAT shape, texture, and ap-pearance in a subsample of 580 participants (mean age = 68.6 y),from 3 different ethnic groups, showed that 30 shape modes explained95% of the VAT shape variance, 40 modes for texture (grayscalevariance), and 30 modes for appearance (combination of the shapeand texture variances). Shape, texture, and appearance PCs remainedin the models (P < 0.05) even after inclusion of the covariates. Themodel including VAT-texture PCs had a better R2 than the models forshape or appearance (0.41 vs. 0.38 and 0.38, respectively). These R2srepresent a slight improvement over the prediction of MetSx using onlydemographic, body size, and body adiposity variables, including VATamount (R2 = 0.33).

Conclusions: The shape and distribution of visceral fat in theabdomen may improve the prediction of cardiometabolic risk, evenafter considering total VAT amount. Further work in this projectwill include more participants, additional race or ethnic groups, andadditional landmark points to refine the models.

Funding SourcesFunded by NCI-P01CA168530 “Obesity, body fat distribution, and

cancer risk in theMultiethnic Cohort” andNIH/NIDDKR01DK109008“Optical Body Composition and Health Assessment”.


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FIGURE 1 A 55-landmark point model was used to define the abdominal structures. The triangles used for visceral adipose tissue texturewarping are shown.

Nonlinear Models Best Characterize the Relationship betweenDietary Cholesterol Intake and Circulating Low-Density Lipopro-tein Cholesterol Levels (P10-130)

Melissa J Vincent,1 Bruce Allen,2 Kevin C Maki,3 Orsolya MPalacios,3 and Lynne T Haber1

1University of Cincinnati; 2Independent Consultant, Chapel Hill,NC; and 3Midwest Biomedical Research/Center for Metabolic andCardiovascular Health, MO

Background: Elevated LDL cholesterol is a major contributor tocardiovascular disease risk, and dietary recommendations focus onlowering intakes of saturated fatty acids (SFAs), trans-fatty acids (TFAs)anddietary cholesterol, all of which tend to raise circulating LDL choles-terol concentrations. Higher intakes of poly- and monounsaturatedfatty acids (PUFAs and MUFAs, respectively) have been found to lowercirculating LDL cholesterol concentrations.

Objectives: The objective of this study was to use flexible dose-response models and data from randomized dietary intervention trialsto characterize the relation between dietary cholesterol intake andcirculating LDL cholesterol, while controlling for SFA, TFA,MUFA, andPUFA intakes using coefficients from the Mensink and Katan equation(2003).

Methods: A literature search (1965–2016) identified 3616 abstracts,of which 55 publications met all inclusion criteria and no exclusion

criteria. Data from the 55 publications were extracted and usedfor a Bayesian meta-regression analysis. The linear, the low-doselinear Michaelis-Menten (MM), and the nonlinear Hill models wereevaluated. Bayesian updating was based on log-likelihoods, defined interms of themagnitude and variability of changes in lipid concentration.An additional parameter was considered in the models to mediate theeffect of baseline cholesterol intake.

Results: A dose-related trend was observed between dietarycholesterol intake and LDL cholesterol values. The results were bestdescribed by the nonlinear MM and Hill models when assessed acrossthe full spectrum of dietary cholesterol intakes studied (0–1500 mg/d).Baseline cholesterol intake was not significantly associated with changein LDL cholesterol for any of the 3 models employed. Mean predictedchanges in LDL cholesterol for an increase of 100 mg/d of dietarycholesterol were 1.90, 4.46, and 4.58 mg/dL for the linear, MM, and Hillmodels, respectively. Corresponding values for an increase of 300 mg/dwere 5.70, 7.76, and 8.06 mg/dL for the linear, MM, and Hill models,respectively.

Conclusions: These results indicate that the relation betweendietary cholesterol intake and LDL cholesterol concentrations are bestcharacterized by nonlinear models and that baseline cholesterol intakewas not a significant predictor of the LDL cholesterol response to achange in dietary cholesterol.

Funding SourcesFunded by American Egg Board-Egg Nutrition Center, Park Ridge,

IL.


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Dietary Fat Influences Epicardial Adipose Tissue Fatty AcidComposition in the OssabawMiniature Pig (P10-131)

Maura E Walker,1 Nirupa Matthan,2 Gloria Solano-Aguilar,3Audrey Goldbaum,2 Sukla Lakshman,3 Huicui Meng,3 SaebyeolJang,2 Joseph F Urban,2 Stefania Lamon-Fava,2 and Alice HLichtenstein2

1Tufts University Friedman School of Nutrition Science and Policy,MA; 2Jean Mayer USDA Human Nutrition Research Center on Aging,MA; and 3USDA, ARS, Beltsville Human Nutrition Research Center,MD

Background: Epicardial adipose tissue (EAT), a visceral fat depotbetween the heart and the pericardium, is thought to promote thedevelopment of coronary artery disease (CAD).However, little is knownabout the influence of dietary fat and statin therapy on EAT fatty acid(FA) composition.

Objective: Our objective was to examine EAT FAs in Ossabaw pigsfed a heart-healthy diet (HHD) or Western diet (WD), with or withoutatorvastatin (S).

Methods:A total of 30Ossabawpigswere randomized into 4 groups,HHD, HHD + S, WD, and WD + S, and fed the respective isocaloricdiets for 6 mo. The diets were matched for macronutrient compositionbut differed in type of dietary fat (unsaturated vs. saturated) andcarbohydrate (whole vs. refined). Pigs fed the HHDwere supplementedwith fish oil [1 g of Epanova, with 550 mg of eicosapentaenoic acid(EPA) + 200 mg of docosahexaenoic acid (DHA)] 3 times a week.EAT adjacent to the left anterior descending coronary artery wascollected and its FA composition was determined by GC. Desaturaseenzyme activities were estimated using the following FA ratios: stearoyl-CoA-desaturase (SCD1; 161n–7/16:0), SCD2 (18:1n-9/18:0), delta-6-desaturase (D6D; 20:3n–6/18:2n–6), and delta-5-desaturase (D5D;20:4n–6/20:3n–6). The results were analyzed by 2-way ANOVA.

Results:There was no significant effect of S on EATFA composition.Relative to the HHD, the WD-fed pigs had higher proportions of totalSFA and trans-FAs, including palmitic (16:0), stearic (18:0), vaccenic(18:1n–7t), and conjugated linoleic (18:2 CLA) acids (all P < 0.01).There were no significant differences in total monounsaturated fattyacids, but pigs fed theHHDhad higher proportions of oleic acid (18:1n–9) (P< 0.01). Pigs fed theHHDalso had higher proportions of total n–6polyunsaturated fatty acids (PUFAs) and n–3 PUFAs, including linoleic(18:2n–6),α-linolenic (18:3n–3), EPA (20:5n–3), anddocosapentaenoic(22:5n–3) acids (all P < 0.01). Estimated activities for SCD1 and D6Dwere higher and SCD2 was lower in pigs fed the HHD compared withthe WD. DHA content and D5D activity were similar between the dietgroups.

Conclusion: These data document, for the first time, that EAT FAcomposition is influenced by dietary fat type. Given the potential effectof FAs and FA-derived lipidmediators on inflammation, themodulationof EAT by dietary fat could influence the association between EAT andCAD.

Funding SourcesFunding for this study was supported by the NHLBI T32 Nutrition

and Cardiometabolic Disorders Pre-doctoral Research Training Grant(NIH T32HL069772-15).

Associations between Epicardial Adipose Tissue Fatty AcidComposition and Gene Expression in the Ossabaw Miniature Pig(P10-132)

Maura E Walker,1 Nirupa Matthan,2 Gloria Solano-Aguilar,3Audrey Goldbaum,2 Saebyeol Jang,3 Joseph Urban,3 AlekseyMolokin,3 Sukla Lakshman,3 Stefania Lamon-Fava,2 and Alice HLichtenstein2

1Tufts University Friedman School of Nutrition Science and Policy,MA; 2Jean Mayer USDA Human Nutrition Research Center on Aging,MA; and 3USDA, ARS, Beltsville Human Nutrition Research Center,MD

Background: Dietary fat type influences epicardial adipose tissue(EAT) fatty acid (FA) composition inOssabaw pigs. Saturated FAs (SFA)can increase inflammation and some polyunsaturated FAs (PUFA)can decrease inflammation via direct/indirect modification of geneexpression.

Objective: The aim of this study was to examine the associations ofselect EAT SFAs and PUFAs with gene expression in the Ossabaw pig.

Methods: A total of 30 Ossabaw pigs were randomized into 4groups and fed isocaloric amounts of a heart-healthy diet (high inunsaturated fat, whole grain, and fruits and vegetables) or aWestern diet(high in saturated fat, cholesterol, and refined grain), with or withoutatorvastatin, for 6 mo. EAT adjacent to the left anterior descendingcoronary arterywas collected. Gene expressionwas determined byRNAsequencing and FA composition was determined by GC. Pigs fromall groups were pooled and Spearman’s correlation coefficients werecalculated between selected EAT FAs (mol%) and EAT gene expression(rpkm).

Results: The following associations were significant (P < 0.0003).SFAs (total SFA, capric, lauric, palmiticm and stearic acids) hadmoderate (r= 0.2–0.49) positive associationswith IRF7 and IFIT1. SFAsalso had moderate (r = 0.2–0.49) positive associations with PTGS2,but associations were weak (r < 0.2) for capric and lauric acids. Totaln–6 (ω-6) PUFAs, linoleic acid, and n–3 PUFAs [total n–3 PUFAs,α-linolenic, eicosapentaenoic, docosapentaenoic, and docosahexaenoicacids (DHA)] had strong positive (r > 0.50) associations with FFAR4and PPARG. Weak (r < 0.20) to moderate (r = 0.20–0.49) positiveassociations emerged between ALOX5 and all PUFAs. DHA wasnegatively associated with IL-1β (r = 0.35), and capric and lauricacids were negatively and weakly associated with IL-6 (r < 0.20). Nosignificant associations were found between any FAs and TLR2, TLR4,MCP1, and IRF3.

Conclusion: EAT SFAs were positively associated with expression of2 interferon signaling genes (IRF7 and IFIT1) and PTGS2, whereas PU-FAs were positively associated with expression of 2 anti-inflammatorygenes (FFAR4 and PPARG). Associations between proportions of EATSFAs and PUFAs with the expression of genes related to inflammationprovide a link between dietary fat type and EAT inflammation.Changing the dietary fat type represents a potential nutritional strategyto reduce EAT inflammation.


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Funding SourcesFunding for this study was supported by the NHLBI T32 Nutrition

and Cardiometabolic Disorders Pre-doctoral Research Training Grant(NIH T32HL069772-15).

Antioxidant Profile, First-Pass Metabolism and AntiproliferativeCapacity of Fruits withGraded Levels of Phenolic Compounds (P10-133)

Abraham Wall-Medrano,1 Francisco Olivas-Aguirre,2 SandraO Mendoza-Díaz,3 Arely I Cárdenas-Robles,3 Emilio Álvarez-Parrilla,2 RamónRobles-Zepeda,4 GustavoAGonzalez-Aguilar,5and Nina R Martinez-Ruiz2

1Universidad Autonoma de Ciudad Juarez—Instituto de CienciasBiomedicas, Mexico; 2Universidad Autonoma de Ciudad Juarez, Mex-ico; 3Universidad Autonoma de Queretaro, Mexico; 4Universidad deSonora, Mexico; and 5Centro de Investigacion en Alimentacion yDesarrollo, A.C., Mexico

Objective: The aim of this study was to evaluate theantioxidant profile, first-pass metabolism, and antiproliferativecapacity of 3 fruits with graded concentrations of phenoliccompounds.

Methods: Phenolic compounds (PCs) and the antioxidant ca-pacity of globe grape, raspberry, and blackberry were evaluatedby spectrophotometry and HPLC coupled to electrospray ioniza-tion quadropole time-of-flight mass spectrometry (HPLC-ESI-Q-TOF-MS). In vitro bioaccessibility of PC subclasses was followedunder simulated gastointestinal conditions (oral, gastric, intestinal).Real-time ex vivo (gut everted sac) monitoring of first-pass phe-nolic metabolites and PCs’ apparent permeability (Papp) were fol-lowed by differential pulse voltammetry (0–120 min), spectropho-tometry, and HPLC-ESI-Q-TOF-MS (120 min). The antiprolifer-ative capacity of main PCs and metabolites was ascertained byMTT assay.

Results: PC richness/fingerprint (cyanidin or pelargodin derivates,catechin, epicatechin), antioxidant (by DPPH or FRAP assay), and elec-trochemical reactivity and bioaccessibility (total PCs and flavonoids)were ranked in the order red blackberry > raspberry > globe grape.The in vitro stability of anthocyanins was pH-dependent. The Papp

of PCs from globe grape favored their absorption, whereas PC effluxwas favored in raspberry and blackberry. The absorption and ex vivobiotransformation of PCs was fruit-dependent. Quinic acid (raspberryand blackberry), malvidin-3-O-glucoside, and caffeic acid (blackberry)were major metabolites, but they did not show antiproliferativecapacity in normal retinal, breast, lung, or colorectal carcinomacells.

Conclusions: The bioaccessibility, chemical stability, and first-passmetabolism of PCs are fruit richness–dependent, but the anticancerpotential of their metabolites (bioequivalence) is not.

Funding SourcesMexican Council of Science and Technology (CONACyT; project

CB-2015-1/254,063).

Depletion of Prohibitin 1 Maintains a Low Glutathione LevelDespite Treatment of S-Adenosylmethionine and Its Combinationwith Taurine and/or Betaine (P10-134)

Yanran Wen, Min Hyung Kim, Gi Eun Heo, and Kwang Suk Ko

Ewha Womans University, South Korea

Background: Liver cancer is the sixth most frequently diagnosedcancer worldwide, but the second most frequent cause of cancer death.Prohibitin 1 (PHB1), a protein in the mitochondrial inner membrane,is a chaperone protein for proper mitochondrial functions. Liver injury,fibrosis, and hepatocellular carcinoma occur in liver-specific Phb1–/–

mice spontaneously, starting from 3 wk after birth. Sulfur-containingamino acid metabolites, S-adenosylmethionine (SAMe), taurine, andbetaine all have a hepatoprotective effect, via regulation of glutathione(GSH) metabolism.

Objective: The aim of this research was to examine the relationsbetween Phb1 expression, treatments of SAMe, and its combinationwith taurine and/or betaine (SCTB), and hepatic GSH concentration.

Methods: The AML 12 cell line, originated from normal mousehepatocytes, was used in this experiment. Cell viability was measuredto confirm the toxicity of each material. mRNA expression of Phb1 andGSH synthesizing and decomposing enzymes were measured in thisstudy.

Results: All materials had no toxicity. However, Phb1 deficiencylowered cell proliferation compared with the control group. GSHconcentration was increased in the SCTB treatment groups comparedwith the control group. In contrast, there was no difference in GSHconcentration in spite of SCTB treatments in the Phb1 knockdowngroups.Gclc expression had no significant difference in all groups.Gclmand Ggt expression was decreased in both the SCTB treatments andthe Phb1 depletion. Gss expression was decreased only in the SCTBtreatments groups.

Conclusions: Treatments with SAMe, and with its combinationwith taurine and/or betaine, have a hepatoprotective effect throughincreasing the GSH concentration by regulating GSH synthesizing anddecomposing enzyme expression. However, this liver protective effectis limited when PHB1 is reduced in mouse liver. Therefore, Phb1expression has important effects on GSH metabolism.


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FIGURE 1 Phb1 knockdown.

FIGURE 2 GSH concentration.


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FIGURE 3 GSH enzyme.

Non-nutritive Sweetener Depth of Knowledge among UniversityStudents (P10-135)

Ted Wilson, Denzel Atherton, Grace Chamberlin, MirandaSaathoff, Breanne MMurray, Lee Schmalz, and Tisha Hooks

Winona State University, MN

Abstract: Understanding consumer knowledge of non-nutritivesweeteners (NNSs), also referred to as artificial or reduced caloriesweeteners, may be important for understandingNNS effects on dietaryheath. NNSs can be identified by the trade name (TN) on packagingor by the chemical or plant name (CN) on ingredient labels. For thisstudy, NNSs were defined as ingredients or chemicals that give a sweettaste to foods and beverages, have no nutritive benefits, and contain fewor no calories. NNS knowledge was evaluated with a survey that wasdelivered to 1248 university students and completed by 493 respondents(age: 19 ± 2 y; ACT score: 24 ± 3) between 1500 on 11 September and1500 on 13 September 2017. To evaluate NNS familiarity, respondentsprovided a fill-in-the-blank NNS definition and NNS examples frommemory. NNS definition depth was graded based for the presenceof 4 different definition categories [score 0 (none) to 4 (excellent);average 1.1 ± 1.1], which was statistically correlated with prior use ofNNSs, food ingredient label use, and class rank. Respondents provided1.0 ± 1.1 examples of true NNSs (CN or TN), which was correlatedwith prior use of NNSs, food ingredient label use, weight loss attempts,and class rank. After being provided with the NNS definition, a 30-item click-drag-box sorting exercise was completed consisting of arandomized list of 6 pairedNNSs (CN and TN), 6 decoyNNSs (not trueNNSs), 6 nutritive sweeteners (not true NNSs), and 6 items that werefoods (not trueNNSs). Click-drag-box exercise food items and nutritive

sweeteners were incorrectly identified as true NNSs only 0.98 ± 1.45and 1.69 ± 1.67 times. In contrast, TN, CN, and decoy items wereidentified as NNSs 4.92 ± 0.98, 3.89 ± 1.87, and 4.70 ± 1.34 times.Significance with a TukeyHSD (P< 0.0001) was TN>CN, TN> decoyname, and decoy name > CN. Although respondents were able toidentify NNSs by TN, presumably from marketing or media exposure,respondents were more likely to incorrectly identify a decoy name asbeing an NNS than a true NNS identified by CN. Respondents werenot able to provide an in-depth NNS definition or able to name theNNSs typically encountered on food packaging or ingredients labels.If consumers do not know what an NNS is, or if they consume them, itmay be difficult to evaluate how NNSs influence our dietary health.

L-Lysine and L-Arginine Supplementations Alter the Expressionof β-actin, Pancreatic Polypeptide, and Enzymes Involved in Glu-tamine Metabolism in Rats (P10-136)

Chaowu Xiao,1 Caroline Faddoline,2 and Carla Wood1

1Health Canada; and 2Carleton University, Canada

Background: L-Lysine (Lys) can be added up to 1.2% in fabricatedsnack products to reduce the formation of acrylamide during cookingin the United States. Lys competes with L-arginine (Arg) for thesame transporter to enter cells. The potential adverse impacts ofexcess Lys intake are poorly understood, and an upper limit of safeintake remains to be established. Our previous study showed thatsupplementation with Lys in a low-protein (7% casein) diet elevatedserum pancreatic polypeptide (PP), a potential modulator of foodintake, and reduced glutamine (Gln) concentrations in rats. However,the potential mechanism(s) underlying these effects remain unclear.


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Objective: The aim of this study was to investigate the effects of Lysand Arg supplementations on the expression of PP in pancreas, and ofβ-actin, glutamine synthetase (GlnS), and glutaminase (Gls) in muscle,liver, and kidney of rats.

Methods: Male Sprague-Dawley rats at 10 wk of age were fed dietscontaining 7% casein with either 0% (CTL), 1%, 1.5%, or 3% Lys, or 3%Lys + 1.5% Arg, for 1 wk. Food consumption and body weight wererecorded. Blood and tissues were collected from all rats at necropsy.Expression of PP, GlnS, Gls, and β-actin in liver, skeletal muscle, andkidney were determined by Western blotting.

Results: Serum Lys and Arg concentrations increased with dietarysupplementation. The rats fed 3% supplemental Lys had lower bodyweight gain and food intake than CTL, but had no effect on pancreaticPP. Serum Gln concentrations were lowered with ≥1% Lys and werefurther decreased by Arg.

Conclusions: Dietary Lys and Arg may regulate serum Gln viadifferent mechanisms, as Lys suppresses muscle synthesis and Argstimulates renal degradation of Gln. Lys and Arg tissue-dependentlymodulate β-actin expression. Supplemental Lys at 3% in a low-proteindiet caused growth retardation and ≥1.5% Lys showed adverse effectsat cellular level. The physiologic significance of these cellular eventswarrants further investigation.

Funding SourcesHealth Canada.

Elevated Intake of PUFA from an Obesogenic Diet Shifts theLipidomicCompositionof Lewis LungCarcinoma inMice (P10-137)

Lin Yan, Sneha Sundaram, Petr Zacek, Michael Bukowski, AaronMehus, and Matthew Picklo

ARS USDA

Background: Metabolic reprogramming of lipid metabolism isa hallmark of cancer. Diet-induced obesity enhances pulmonarymetastasis of Lewis lung carcinoma (LLC) in mice.

Objective: The objective of this study was to determine the impactof feeding a high-fat, obesogenic diet upon the lipidomic compositionof LLC in mice.

Methods: LLC primary tumors were obtained from mice fed acontrol AIN-93Gdiet or a high-fat diet (HF), with 16%or 45%of energyfrom soybean oil, respectively. Four lipid species [phosphatidylcholines(PC), phosphatidylethanolamines (PE), ceramides (CER), and triacyl-glycerols (TAG)] were quantified using a shotgun mass spectrometryapproach.

Results: Concentrations of 51 PCs were different in tumors fromthe mice consuming the 2 diets. Six of these PCs were increased and2 were decreased ≥2-fold by the HF diet. There were 24 PEs thatwere responsive to the different diets; one of them, PE(18:2_20:2),was 2-fold greater in HF tumors than in the controls. The content ofCER(d18:1/24:2) was 49% higher and CER(d18:1/24:1) was 33% lowerin HF tumors than in the controls. We identified 62 TAG structuresranging from 46:0 to 60:15, 12 of which were increased and 3 decreased≥2-fold by theHF diet. Hierarchical clustering heatmap analysis of PCsand PEs and neutral loss analysis of TAGs showed that polyunsaturatedfatty acid lipid species were higher in tumors from mice fed the HFdiet and that monounsaturated fatty acid (MUFA) lipids were higherin tumors from the control mice. Although there was a reduction in

MUFAs, there were no changes in the de novo lipogenesis enzymes fattyacid synthase, stearoyl-CoA desaturase 1, or acetyl CoA carboxylase 1in tumors from HF-fed mice compared with the controls.

Conclusions:These results indicate that the lipidomic profile of LLCprimary tumor reflects the fatty acid composition of the diet. It suggeststhat lipidomic alteration of the primary tumor by an obesogenic dietmay contribute to the aggressiveness of LLC and its metastasis to thelungs.

Funding SourcesThis work was supported by the USDA, Agriculture Research

Service, Research Project 3062-51,000-050-00D.

Foodomics Database: A New Tool for Estimating MolecularProfiles of Nutrient Intake during Precision Ketogenic Therapy(P10-138)

Lujia Yang and Peggy Borum


Objective: Precision ketogenic therapy (PKT) is a high-fat, low-carbohydrate, adequate-protein diet that is prescribed using themacronutrient information on nutrition facts labels (NFL) of brand-name products. The purpose of this study was to create a foodomicsdatabase to estimate themolecular compounds (individual amino acids,fatty acids, and carbohydrates) of brand-name products and to evaluatepatients’ nutrient intake.

Methods: The units of the molecular compounds in USDA SR 28are converted from g/100 g of food to g/1 g of the macronutrient,resulting in the pattern of molecular compounds in correspondingmacronutrients. Similar types of NFL products share the same baseproduct calculated from the mean of nutrients of all related genericproducts. By using the amount of macronutrients listed on the NFL fordifferent brand-name products and for the base product, the amountsof molecular compounds are listed in the foodomics database. Daily di-etary intake data of 2 patients (KG0232 andKG0222), who receivedPKTfrom2015 to 2016, were used to examinewhether this database could beused for documenting the amount of molecular compounds consumed.

Results: The foodomics database was created for brand-nameproducts using very recent NFL data from products collected in Floridaand used to document the amount of molecular compounds consumedby patients (Table 1). Data from these 2 patients illustrate that use of thefoodomics data permits researchers to address questions about whethertube-fed and G-tube–fed patients on PKT have a similar or differentintake of: total branched-chain amino acids; profile of branched-chainamino acids; medium-chain saturated fatty acids versus long-chainsaturated fatty acids; ratio ofω-6 (n–6) toω-3 fatty acids; and percentageof ω-6 and ω-3 fatty acids that are 18 carbons versus >18 carbons long.

Conclusions: Researchers can use the foodomics database toestimate the amount of molecular compounds in different brand-nameproducts and to estimate and document the molecular compounds in apatient’s daily intake.

Funding SourcesResearch reported in this publication was supported by the Uni-

versity of Florida Clinical and Translational Science Institute, whichis supported in part by the NIH National Center for AdvancingTranslational Sciences under award number UL1TR001427, and also bythe University of Florida Foundation (KetoGator, Fund ID: 01,8555).


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TABLE 1 Molecular compounds intake of patients who receivedPKT

The Differences of Dietary Intakes and Dietary Habits in KoreanLean and Obese Diabetes Patients (P10-139)

Jung-Eun Yim, Ha-Neul Choi, and Su-Mi Suk

Changwon National University, South Korea

Background: The majority of patients with type 2 diabetes (T2D)are obese or overweight. The percentage of lean T2D patients in Koreais higher than in Western countries, but the percentage of obese T2Dpatients is rapidly increase in Korea. Lean diabetes patients might havehigher total mortality and rapid beta cell failure. Poor dietary habits andlifestyle are the major causes of T2D.

Objective: The aim of this study was to characterize and comparethe dietary intake and dietary habits of Korean lean and obese T2Dpatients.

Methods: A total of 36 T2D patients were recruited. These wereclassified as obese T2D [body mass index (BMI): ≥25 kg/m2] and leanT2D (BMI:<25 kg/m2). We used 3-d food record evaluation to analyzedietary intake using computer aided nutritional program (version 5.0),and eating habits and health-related behaviors were investigated. Thecollected data were analyzed statistically using SPSS statistics version24.

Results:Obese T2D tend to have a high percentage of family historyof disease compared with lean T2D. Comparing the health-relatedlifestyles of the 2 groups, lean T2D had higher intakes of functionalfoods, non-smoking, and exercise than obese T2D. The intakes ofenergy, fat, cholesterol, and protein in the obese T2D tended to behigher than in the lean T2D. However, the intakes of vitamin C and

α-linolenic acid were significantly lower in obese than in leanT2D. As for the dietary habits of both groups, the nonobese T2D atebreakfast more regularly than the obese T2D. The lean T2D had longermealtimes than the obese T2D. The obese T2D had a higher frequencyof overeating and snacking than the lean T2D. The obese T2D preferredto eat salty, spicy, and greasy foods. Therefore, the overall eating habitsof the lean T2D tend to be better.

Conclusions: The group of lean T2D patients showed a tendency tohave a better lifestyle and eating habits than the obese T2D, leading tohealthy intake of nutrients.

Funding SourcesAn NRF grant funded by the Korea Government (NRF-

2016R1D1A1B03935660).

Plasma Metabolomics Profiles Reveal Novel Metabolites in EarlyPregnancy in Association with Gestational Diabetes Risk (P10-140)

Cuilin Zhang,1 Yen-Chen Anne Feng,2 Oliver Fiehn,3 MichaelTsai,4 Yeyi Zhu,5 Paul Albert,6 and Liming Liang7

1National Institute of Child and Human Development, NIH;2Massachusetts General Hospital; 3UC Davis, CA; 4University ofMinnesota; 5Kaiser Permanente Northern California; 6NCI, NIH; and7Harvard Chan School of Public Health, MA

Objective: Although metabolomics may shed light on the patho-physiology of glucose intolerance in pregnancy, prospective andlongitudinal studies on their roles in the development of gestationaldiabetes (GDM), a common pregnancy complication that has adverse


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implications onwomen and the index offspring, are sparse.We aimed toprospectively examine metabolomics (both targeted and nontargeted)and GDM risk.

Methods: A matched case-control study of 107 GDM and 214non-GDM women was conducted in a multiracial cohort, the NICHDFetal Growth Studies. GDM diagnosis was based on the Carpenter& Coustan Criteria. A total of 22 amino acids (AAs) were quantifiedby a Hitachi-8900 Amino Acid Analyzer using plasma collected atgestational weeks (GW) 10–14, 15–26, 23–31, and 33–39. In addition,331 primary metabolites were quantified by GC-time of flight-MS. Thepresent report focuses on findings in early pregnancy (GW 10–14).Adjusted odds ratios (aORs) and 95%CIs ofGDMrelated tometabolites[in quartiles (Qs)] were estimated using conditional logistic regressionafter adjusting for age, prepregnancy body mass index, and other GDMrisk factors. Multiple testing was corrected using false discovery ratecontrol.

Results: Among the AAs, alanine concentrations in GW 10–14were significantly higher in GDM women than in controls (mean:30.5 vs. 27.6 µmol/dL) and were positively related to GDM risk; aORsacross increasing Qs were 1.00, 1.69, 2.86, and 3.05 (P-trend = 0.02).By contrast, asparagine and glycine concentrations were significantlylower in GDM women than in controls, and were inversely relatedto GDM risk; aORs across increasing Qs were 1.00, 0.92, 0.74,and 0.48 (P = 0.045) for asparagine and 1.00, 0.42, 0.36, and 0.24(P -4).

Conclusion: Our study revealed several novel metabolites that maybe implicated in the early pathogenesis of GDM. These findings mightprovide new etiologic insight into the development of GDM.

Funding SourcesFunded by an intramural research program, EuniceKennedy Shriver

National Institute of Child Health and Human Development, NIH.

Grain Fortification with Vitamins and Iron May Be a MajorCause of the Prevalence of Obesity and Diabetes in the United States(P10-141)

Yiming Zhou,1,2,3 Nana Chen,4 Da Li,5 Wuping Sun,4 and Shi-Sheng Zhou4

1Harvard Medical School, MA; 2Brigham and Women’s Hospital,MA; 3Broad Institute, MA; 4Medical School of Nagoya University,Japan; and 5Shengjing Hospital of China Medical University

Background: The United States was the first country to fortifygrains with fat synthesis–promoting B vitamins (thiamin, riboflavin,and niacin) and iron, which it initiated in the late 1930s. It is also thefirst country to experience an obesity and diabetes epidemic, especiallyin its southern states, where grain fortification is mandatory. However,whether grain fortification may play a causal role in the prevalence ofobesity and diabetes remains to be determined.

Objective: The objective of this study is to investigate whether grainfortification plays a causal role in the prevalence of obesity and diabetes.

Methods: The present study investigates the relation between grainfortification and the prevalence of obesity and diabetes by Pearson’scorrelation analysis. Per capita nutrient consumption data from 1909to 2010 were obtained from the USDA Economic Research Service, andobesity and diabetes prevalence data were obtained from the Center forDisease Control from 1958 to 2015.

Results: Grain fortification has increased the levels of per capitaconsumption of thiamin, riboflavin, niacin, and iron, from ∼1.4, 1.9,17, and 12.5 mg/d, respectively, in the early 1930s to peak values of 3.1,3.2, 35, and 25.1 mg/d in the early 2000s. The increased consumptionof thiamin, riboflavin, niacin, and iron has been accompanied by an∼30% increase in US per capita calorie consumption (from 3250 kcal/din the 1930s to 4200 kcal/d in the early 2000s). The prevalence of bothobesity and diabetes is strongly correlated with the increased US percapita consumption of thiamin, riboflavin, niacin, and iron, with timelags of ∼10 and 23 y, respectively (all Pearson’s correlation coefficientr > 0.91).

Conclusions: Given that our previous studies have shown thatexcess nicotinamide (a niacin form most commonly used in grainfortification) may increase the concentration of reactive oxygen species,cause insulin resistance, and alter the body weight in rats and humans,we think that grain fortification with vitamins and iron may be a majorcause of the increased prevalence of obesity and diabetes in the UnitedStates.

Funding SourcesNational Natural Science Foundation of China.

A Prospective and Longitudinal Study of Plasma Phospholipidn–3 and n–6 PUFAs in Relation to Gestational Diabetes Risk (P10-142)

Yeyi Zhu,1 Mengying Li,2 Shristi Rawal,3 Stefanie N Hinkle,2Jing Wu,4 Jagteshwar Grewal,2 Huixiang Yang,5 Michael Y Tsai,6Assiamira Ferrara,1 and Cuilin Zhang2

1Kaiser Permanente Northern California Division of Research;2Eunice Kennedy ShriverNational Institute of ChildHealth andHumanDevelopment; 3Rutgers School of Health Professions, NJ; 4Glotech Inc.;5Peking University First Hospital, China; and 6University of Minnesota

Background: Despite recommendations on dietary intakes ofpolyunsaturated fatty acids (PUFAs) for cardiometabolic health, data onn–3 (ω-3) and n–6 PUFAs in relation to diabetes risk remain debated.Further, data are lacking in pregnant women.

Objective: The purpose of this study was to prospectivelyinvestigate individual plasma phospholipid n–3 and n–6 PUFAsthroughout pregnancy in relation to risk of gestational diabetes(GDM).

Methods: Within the NICHD Fetal Growth Studies-SingletonCohort of 2802 pregnancies, 107 GDM cases were ascertained bymedical record review and matched to 214 non-GDM controls onage, race or ethnicity, and gestational week (GW) at blood collection.Individual plasma phospholipid n–3 and n–6 PUFA concentrationswere measured by GC at GW 10–14, 15–26, 23–31, and 33–39.Conditional logistic regressionwas used, adjusting formajor risk factorsfor GDM.

Results: Overall, n–3 PUFAs decreased slightly and n–6 PUFAsdid not change appreciably across gestation. Among the n–3 PUFAs,docosapentaenoic acid (22:5n–3) at GW 15–26 was significantly andinversely associated with subsequent risk of GDM [adjusted odds ratiocomparing the highest vs. lowest quartile (aORQ4-Q1): 0.29 (95% CI:0.10, 0.85); P-trend = 0.048]. Among the n–6 PUFAs, γ -linolenic acid(18:3n–6) and dihomo-γ -linolenic acid (20:3n–6) at GW 10–14 werepositively associated with risk of GDM [aORQ4-Q1: 2.53 (95% CI: 1.12,


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5.68) and 3.42 (1.37, 8.54), respectively, both P-trendQ4-Q1 = 0.30 (0.11,0.83), P-trend = 0.015]. Further, the per SD increase in the PUFAn–6:n–3 ratio at GW 15–26 was related to a 1.64-fold (1.09, 2.46)increased risk of GDM. Jointly, high concentrations of (≥median) n–3PUFAs and low concentrations of (<median) n–6 PUFAs were relatedto a 64% [aORQ4-Q1: 0.36 (0.14, 0.95)] decreased risk of GDM comparedwith women with low n–3 PUFA concentrations and high n–6 PUFAconcentrations.

Conclusions:Our novel findings highlight the potentially importantdifferential roles of individual n–3 and n–6 PUFAs and the PUFA n–6:n–3 ratio during early to mid-pregnancy for GDM development. Ourfindings may underlie distinct nutritional, metabolic, or physiologicprocesses, and inform potential prevention strategies.

Funding SourcesNICHD intramural funding; YZ is also supported by

3K12HD052163-18S1.


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Energy and Macronutrient Metabolism

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