Health and Nutrition

ABSTRACTS 2019 AOCS ANNUAL MEETING & EXPO May 5–8, 2019

HEALTH AND NUTRITION

H&N 1: Lipids and Inflammation Chair: Eric Murphy, University of North Dakota, USA

Increase in Plasma Ganglioside Content is significant effect of ganglioside on bowel Associated with Improved Quality of Life in symptoms or social function. Inflammatory Bowel Disease John J. Conclusions. Daily consumption of Miklavcic*1, Glen Shoemaker2, Kareena L. ganglioside increases plasma ganglioside Schnabl3, Alan BR Thomson4, Vera Mazurak2, concentration and constitutes a viable agent for and M Tom Clandinin5,1Chapman University, treatment of IBD or for potential treatment of USA; 2University of Alberta, Canada; other disorders in which altered ganglioside 3Laboratory Medicine and Pathology, University content has been implicated. of Alberta, Canada; 4Gastroenterology, Western University, Canada; 5Centre for Health and FABP1 and Its Emerging Role in Systemic Nutrition, University of Alberta, Canada and Brain Endocannabinoid Systems. Eric Dietary ganglioside has been shown to Murphy, University of North Dakota, USA decrease inflammation, improve gut-barrier Pending. function, and prevent infection by pathogenic micro-organisms. Are Oxidized Linoleic Acid Metabolites Objective. To determine if dietary Bioavailable? Ameer Taha*, University of ganglioside is bioavailable in human California, Davis, USA participants and whether ganglioside consumption improves clinical outcomes in Background: Omega-6 linoleic acid (LA, 18:2n-6) inflammatory bowel disease (IBD). is a precursor to oxidized linoleic acid Methods Used. Healthy control participants metabolites (OXLAMs) which regulate multiple and patients with IBD were recruited for a biological processes. In vivo, OXLAMs can be double-blind, randomized, placebo-controlled synthesized enzymatically or non-enzymatically study. Participants were allocated to receive from LA. Recently, our group characterized their either a 1.0 g milk fat fraction with or without abundance in plant oils. However, little is ganglioside (80% GD3; 20% GM3) daily for 8 known about their bioavailability. weeks. Blood was drawn to measure plasma Objective: The goal of this study is to determine ganglioside content at weeks 0, 2, 4, 6 and 8. the bioavailability of dietary OXLAMs. Participants had bowel symptoms, emotional Methods: Two rodent experiments were health, systemic symptoms, and social function performed to address the question of whether assessed by the Quality of Life in IBD OXLAMs are bioavailable or not. In Experiment Questionnaire. 1, mice were fed for 8 weeks a low LA diet (4% Results. Level of plasma ganglioside GD3 energy), high LA diet (17% energy) or a low LA rapidly increased by 42% after two weeks and diet (4% energy) supplemented with thermally by 49% after eight weeks (P<0.05). Level of oxidized corn oil containing OXLAMs. Plasma ganglioside GM3 also increased by 8% after and brain OXLAM concentrations were eight weeks (not significant). Plasma levels of quantified by UPLC-MS/MS. In Experiment 2, GM3 and GD3 did not change between baseline rats were gavaged with deuterated 13- and study conclusion in subjects that consumed Hydroxyoctadecadienoic acid (13-HODE) and placebo. Dietary ganglioside improved tracer incorporation in plasma measured by emotional health by 39% (P<0.01) and systemic UPLC-MS/MS at 90 minutes. symptoms by 36% (P<0.02) over the course of Results: In Experiment 1, OXLAM consumption study in patients with IBD. There was no (through the low LA + OXLAM diet) did not alter

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plasma or brain OXLAM concentrations kidney diseases revealed increased levels of compared to the low LA diet. The high LA diet lysophospholipid and phospholipase A2, and increased plasma and brain OXLAM increased activities of cyclooxygenases (COX). concentrations compared to the low LA diet. In COXs are one of 3 major enzyme pathways that Experiment 2, labeled 13-HODE was detected in result in the production of oxylipins. plasma one hour post-gavage (n=1), suggesting Subsequent studies showed that drugs and absorption. Ongoing analysis will seek to dietary interventions that reduced disease confirm this finding in two other gavaged rats. progression also reduced the production of Conclusion: Dietary OXLAMs may be absorbed select COX derived oxylipins in these pediatric through the GI tract but accumulation in plasma forms of the disease. With the advent of and brain appear to be minimal. Thus, in vivo technologies that enabled the lipidomic analysis OXLAM concentrations are regulated by dietary of >100 oxylipins by HPLC/MS/MS, we then LA. showed that the COX derived oxylipins were consistently elevated in both pediatric and adult Oxylipins in Kidney Inflammation Harold M. forms of cystic kidney diseases. Further, these Aukema*, University of Manitoba, Canada studies showed that these bioactive lipids were Inflammation plays an important role in the elevated early in the disease, and when the progression of kidney diseases. Oxylipins are disease had only progressed minimally. We lipid mediators that are involved in normal then showed that selective COX2 inhibition kidney physiology and response to physiological reduced disease progression by blocking the challenges, but when present during kidney production of oxylipins in a mouse model of the disease they can contribute to inflammation most common form of cystic kidney disease. and worsening of disease. Cystic kidney Selectively blocking COX2 derived oxylipins diseases are the major cause of inherited kidney therefore provides a potential therapeutic diseases world-wide, but treatment options for intervention that reduces inflammation and this group of kidney disorders is very limited. disease progression in these kidney diseases. Our early studies with forms of pediatric cystic

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EAT 1.1 / H&N 1.1: Structural Determinates of the Metabolic Response for Lipids Chairs: Michael Rogers, University of Guelph, Canada; and Pamela Hutton, Bunge Loders Croklaan, USA

Replacement of Saturated Fat with composition and micronutrient levels result in Unsaturated Fats from Different Food Sources: the varying health effects of different vegetable Implications for Cardiovascular Risk Kristina S. oils. However, the mechanisms underlying this Petersen*, The Pennsylvania State University, phenomenon have largely remained elusive. USA Foodomics is a discipline that studies the food Current dietary guidance for prevention and and nutrition domains through the application management of cardiovascular disease includes of advanced omics technologies to improve recommendations to replace saturated fats consumer’s well-being, health, and confidence. with polyunsaturated fatty acids (PUFA) or The objective of this study was to investigate monounsaturated fatty acids (MUFA). This the health effects of different vegetable oils by presentation will provide an overview of foodomics analysis. The physicochemical current research focusing on how replacement properties and oxidative stability of seven of saturated fat with MUFA and PUFA from commercial vegetable oils were evaluated, different food sources modulates lipids and including soybean oil, rapeseed oil, corn oil, lipoproteins, blood pressure, and vascular sunflower seed oil, rice bran oil, peanut oil and health. Our lab has shown that replacement of tea seed oil. And then SD rat were fed with high saturated fat with walnuts, or vegetable oils rich fat diet contained different vegetable oil for 12 in MUFA and PUFA improves lipids and weeks. Metabolites in serum and liver tissue lipoproteins, and blood pressure. Furthermore, were measured using liquid chromatography– data from a randomized, crossover controlled and gas chromatography–mass spectrometry, feeding study showed canola oil and high-oleic and multivariate statistical analysis was acid canola oil improved lipids and lipoproteins performed by orthogonal projections to latent compared to an oil blend higher in saturated structures discriminant analysis. The finding fat. Based on these data and other recent work showed vegetable oils, especially tea seed oil the magnitude of lipid and lipoprotein lowering and rapeseed oil, decrease the liver lipid expected with different dietary replacement of metabolism disorder in the high fat diet treated saturated fat will be explored. In summary, the mice. This study provided the foundation for research presented will describe the effect of quality, nutrition and safety evaluation of different fatty acids within the food matrix on vegetable oils and reasonable reference for cardiovascular risk and how we can assist consumers. consumers to choose foods and dietary patterns that are heart healthy. Musseling-up Program: Review of Greenshell Mussel Bioactive Lipids and Role in Foodomics Insights in the Health Effects of Inflammation Management and Joint Health Vegetable Oil YongJiang Xu*1, Chen Cao1, Matthew R. Miller*1, Marlena C. Kruger2, Fran Zhaojun Zheng1, and Yuanfa Liu2, 1Jiangnan M. Wolber2, Hong Tian Hong Tian3, Parkpoon University, China; 2School of Food Science and Siriarchavatana2, and Saima Rizwan4, Technology, State Key Laboratory of Food 1Cawthron, New Zealand; 2Massey University, Science and Technology, Jiangnan University, New Zealand; 3Sanford Ltd, New Zealand; China 4Riddet Institute, New Zealand Vegetable oil plays an important role in Lipid extracts from New Zealand’s iconic daily diet. The differences of fatty acid Greenshell mussel (GSM) are the world’s most

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expensive nutritional oil (~$2000 USD/kg). 2USDA, ARS, Grand Forks Human Nutrition Traditional use of GSM by coastal Maori has Research Center, USA been associated with improved joint health and Objective: Dietary carotenoids are mainly a number of studies have examined the anti- sourced from fruits and vegetables. The inflammatory effects of GSM extracts. bioavailability of carotenoids is dependent on Objectives This review looks at outputs of dose, quantity and dispersion, and presence of Musseling up program a 3-year program under other nutrients in the diet, specifically fat. the New Zealand National Science Challenge However, there is a gap in research on whether “High Value Nutrition” by a) identifying, specific fatty acid classes affect serum β- discriminating and verifying GSM active carotene concentrations. Our primary objective ingredients; b) producing novel GSM was to assess the association between serum β- food/extract products; c) determining the carotene concentrations and reported intake of mechanism and efficacy in pre-clinical and specific fatty acid classes, utilizing data the clinical trials. Methods We have determined What We Eat in America (WWEIA)/National GSM lipid content over seasons, developed Health and Nutrition Examination Surveys rapid near infrared analytical methods and (NHANES). Methods: Data from 9,182 male and developed novel food applications. Efficacy of female participants 20-85 years of age in the GSM was tested 1) in-vitro analysis in NHANES 2003-2006 nationally representative, macrophage, cartilage osteoclast and pre- cross-sectional survey were analyzed to osteoblast cell lines, 2) in-vivo in female estimate the relationships between serum β- Sprague-Dawley rats with obesity-induced carotene concentrations and reported osteoarthritis and 3) in a clinical trial on saturated (SFA), monounsaturated (MUFA), and bioavailability of the lipids fraction. Results We polyunsaturated (PUFA) fatty acid intakes. Due have determined the variation in GSM with to skewing, we log transformed serum β- female spring mussels having the highest lipid carotene. Multiple linear regression estimated content. Novel stable GSM rich food ingredients log(serum β-carotene) based on total reported have been made through emulsification and fatty acid intakes adjusted for age, sex, and spray drying techniques. The inclusion of GSM ethnicity. Results: Mean and standard error (SE) in the rat diet significantly reduced blood levels was 2.51±0.85µg/dL for log(β-carotene). Mean of a cartilage degradation biomarker in rats In and SE for SFA were 26.72±16.9g, MUFA were vitro, a non-polar lipid extract of GSM 30.40±18.4g, and PUFA were 17.22±11.6g. β- significantly reduced osteoclast differentiation carotene concentrations were weakly and in a dose-dependent manner. Clinical trials have inversely associated with fatty acid classes: SFA determined the extent of the bioavailability of (r= -0.15, p the lipid fractions in 4 different formats. Conclusions GSM food products, food Oleogelation of Emulsified Oil Delays in vitro ingredients and extracts provide exciting Intestinal Lipid Digestion Dérick Rousseau*, opportunities for improving joint health and Ryerson University, Canada inflammation management. The influence of oleogelation on the in vitro digestion of oil-in-water (O/W) emulsions was Serum β-carotene Concentrations is Inversely investigated. Rice bran wax (RBX) was used as Associated with Reported Fatty Acid Intake in an oleogelator at concentrations of 0, 0.25, 0.5 U.S. Adults Ambria Crusan*1, Marla Reicks1, and 1 and 4 wt% of the emulsions, respectively. Susan K. Raatz2,1University of Minnesota, USA; O/W emulsions containing 1 wt% whey protein

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and 20 wt% oil were prepared via hot buffer agent (e.g., Mg(OH)2 or CaCO3). This homogenization, and characterized by rheology, buffer was selected because it is insoluble static light scattering, confocal/polarized light under alkaline conditions, but soluble under microscopy, X-ray diffraction and differential acidic conditions, and can therefore maintain a scanning calorimetry. The evolution in particle neutral pH inside the microgels when they are size distribution, structural changes during oral, dispersed in a low pH solution (such as acidic gastric and intestinal digestion, and free fatty gastric fluids). A quantitative ratiometric acid (FFA) release during intestinal digestion method based on laser scanning confocal were all investigated. All emulsions were microscopic imaging was also developed to map generally kinetically stable during the the microclimate pH inside microgels. The experimental timeframe (4 weeks), with the image method was used to detected the pH exception of the 4 wt% emulsion, given the change of the microgels with or without the eventual appearance of large RBX crystals (3-5 encapsulation of buffer agent during digestion µm) in the droplets. The rheology and thermal process. It was indicated that the initial pH behaviour of the emulsions, as well as the SBO- distribution was relatively uniform within the RBX blends used for each emulsion, buffer-free microgels, being in the range from demonstrated that addition of RBX led to the pH 6.8-7. For the buffer-loaded microgels, the formation of rigid oil droplets. During both center pH was slightly higher than the edge gastric and intestinal digestion, oil droplet with the range from 7.2-7.6. After simulated coalescence occurred in all emulsions, and stomach digestion, the microclimate inside the intestinal digestion led to oil droplet microgels changed to acid condition. aggregation during the early stages of intestinal Conversely, the pH value within microgels digestion. The FFA release profiles showed that containing buffer agents was fairly similar to oleogelation of emulsified oil delayed intestinal that of the initial value. The activity of loaded lipid digestion. This effect was enhanced with bioactives (i.e., enzymes, insulin or probiotics) increasing RBX concentration up to 1 wt% due were successfully reserved after encapsulation to the increased oil droplet rigidity. With a within the buffer-loaded microgels during the further increase to 4 wt%, the rate of lipid digestion process. Our results could provide digestion increased, which was ascribed to the valuable information for the design of instability of the emulsions containing 4 wt% microclimate pH mapping technique and RBX during digestion caused by large RBX development of nutraceutical delivery systems crystals. for acid sensitive bioactive ingredients.

Encapsulation, Protection and Controlled Relea The Role of Emulsifiers in Lipid Digestion of Oil- se of Nutraceuticals using Biopolymer Microgel in-Water Emulsions Michael Rogers*, Natalie Zipei Zhang*, and D. Julian J. Ng, Saeed M. Ghazani, Peter Chen, Alejandro G. McClements,University of Massachusetts Marangoni, Amanda Wright, and Douglas Amherst, USA Goff,University of Guelph, Canada Microgels are one promising colloidal Fat digestion significantly influences health. delivery systems widely used for the Both food composition and food structure encapsulation and release of bioactive influence lipid digestion kinetics and the ingredients. In this study, a novel self-regulating bioavailability of components they contain. hydrogel microgels were fabricated that Thus, we need to understand how ALL levels of contained acid sensitive compounds and a food structure (i.e., nano-, micro-, macro- and

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mesoscales) control the lipemic index of foods. pesticide (chlorpyrifos) was studied. The results Since surfactants are ubiquitously found in indicated that the bioaccessibility of processed foods, there is potential benefit to chlorpyrifos (a highly lipophilic pesticide) was design food emulsions with tailored lipid shown to depend on the emulsifier types of co- digestion profiles. Ultimately our aim is to limit ingested excipient emulsion. Highest pesticide the lipemic index of processed foods however, bioaccessibility can be observed for the with the emergence of personalized nutrition excipient emulsion formed with phosphate there are applications where rapid digestion lipid, followed by Tween 80 and WPI. kinetics could be highly beneficial. 200 ± 20 nm, Polysaccharide additions (chitosan, xanthan, β- oil-in-water emulsions, structured using glucan) can significant impact the common chain-length (i.e., C18:0) surfactants bioaccessibility of pesticides that it also (e.g. sn-1, and sn-2 monostearin, Span 60, and depended on the emulsifier type of excipient Tween 60) had their digestion kinetics studied emulsions. Overall, these results suggest that using the TIM-1 simulated gastrointestinal tract. the bioavailability of undesirable pesticides can Lipid digestion rate constants, induction times be controlled by specificity design excipient and bioaccessibility were assessed over 5 hr emulsion. simulated digestions. All parameters are significantly influenced by emulsifier structure, Impact of Indigestible Oils on the creating exciting new opportunities in Bioaccessibility of Vitamin D3 in structuring infant formulas, energy-dense meal Nanoemulsion-based Delivery Systems Yunbing replacements, etc. Tan*1, and D. Julian J. McClements2,1Dept. of Food Science, University of Massachusetts, Bioavailability of Pesticide Residue in Amherst, USA; 2University of Massachusetts Agricultural Products: Impact of Food Amherst, USA Emulsions with Different Surface Properties These is interest in replacing digestible fats Ruojie Zhang*, and D. Julian J. McClements, with indigestible ones to reduce the calorie University of Massachusetts Amherst, USA content of foods. However, utilization of The residue of pesticides in agricultural indigestible oils may have unforeseen products has been a big concern for human nutritional consequences. In this study, the health since high exposure to pesticide could impact of an indigestible oil on the cause numbers of diseases, such as diabetes, bioaccessibility of vitamin D3 (VD) encapsulated neurological disorders, even cancer. The within nanoemulsion-based delivery systems Bioavailability of pesticides residue in human was examined. Four different nanoemulsions body are highly impacted by the food were prepared using different combinations of components that consumed together. Food a digestible oil (corn oil, CO) and indigestible oil emulsions, such as dressings, dips, sauces, and (mineral oil, MO): CO only; MO only; 1:1 CO:MO creams, are commonly co-ingested with fruits system prepared by mixing oils before and vegetables. The purpose of the current homogenization (oil mixture); 1:1 CO:MO study was therefore to examine the potential system prepared by mixing MO and CO impact of co-ingestion of emulsions with nanoemulsions after homogenization (emulsion natural produce on the bioavailability of a mixture). A simulated gastrointestinal tract hydrophobic pesticide. In current study, the (GIT) was used to study the interaction between influence of co-ingestion of food emulsions with pancreatic lipase and the emulsions. The rate of tomatoes on the bioaccessibility of a model free fatty acid release and the VD

– 6 – ABSTRACTS 2019 AOCS ANNUAL MEETING & EXPO May 5–8, 2019 bioaccessibility during 2 h intestine digestion attributed to solubilization of the VD in the decreased in the same order: CO > oil mixture ≈ mixed micelles, followed by their precipitation. emulsion mixture > MO. The digestion of the These results show that lipid digestion, micelle corn oil occurred primarily during the first 30 solubilization, and micelle aggregation all minutes, then gradually increased. Except for impact vitamin bioaccessibility. The presence of the MO nanoemulsion, the bioaccessibility of indigestible oil in the nanoemulsion-based VD increased to a maximum value around 30 delivery systems reduced vitamin minutes of digestion but then decreased during bioaccessibility, but may be useful for giving the following 24 h. This effect might be prolonged release in the colon.

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H&N 2: Brain Lipid Biochemistry Chairs: Eric Murphy, University of North Dakota, USA; and Charles Nider, Abitec Corp., USA

How Docosahexaenoic Acid Enters the Brain: enzymatic esterification. Methods: Post- Consensus, Controversies and Updates Richard mortem pre-frontal cortex from pathologically P. Bazinet*, University of Toronto, Canada confirmed AD subjects (n=21) and unaffected The brain requires a constant supply of controls (n=20) was subjected to lipidomic docosahexaenoic acid from blood to maintain analysis with ultra-high pressure liquid its levels within the brain. Several plasma pools chromatography coupled to tandem mass- have been proposed to supply the brain with spectrometry following separation of brain docosahexaenoic acid, including plasma esterified and unesterified lipid pools with solid lipoproteins lysophosphatidylcholine and phase extraction. Results: Compared to unesterified fatty acids. In this lecture, I will controls, concentrations of several pro-repair review the evidence for each plasma pool lipid mediators esterified to neutral lipids were supplying the brain highlighting consensus, significantly reduced by ~50% in AD patients controversies and remaining questions. While (P<0.05). No significant changes were observed circulating lysophosphatidylcholine has a higher in free or phospholipid-bound pro-repair lipid brain/body partition coefficient than mediators. unesterified docosahexaenoic acid, unesterified Conclusion: This study provides novel docosahexaenoic acid entry into the brain is evidence of reduced esterified pro-repair lipid more rapid. The implications of how plasma mediators in post-mortem AD pre-frontal docosahexaenoic acid-containing cortex. Targeting pro-repair lipid mediator lysophosphatidylcholine and unesterified turnover within neutral lipids may stimulate docosahexaenoic acid contribute to maintaining neuronal repair and resolution of inflammation brain docosahexaenoic acid concentrations will in AD. also be discussed as well as several recent findings. Lipid Binding Proteins and Brain Fatty Acid Uptake and Metabolism: So What is the Role Lipidomic analysis of post-mortem Alzheimer's of Liver- Fatty Acid Binding Protein (FABP1)? Disease pre-frontal cortex reveals changes in Eric J. Murphy*, University of North Dakota, pro-repair lipid pathways Ameer Taha*, USA University of California, Davis, USA Over the past 15 years, my laboratory has Background: Alzheimer’s Disease (AD) is a explored the role of traditional fatty acid progressive brain disorder characterized by binding proteins (FABP) and non-traditional extensive inflammation and neuronal damage. fatty acid binding proteins in facilitating fatty Specialized lipid mediators known to resolve acid uptake and trafficking in the brain. We neuroinflammation and repair damaged demonstrated that heart FABP (FABP3), which is neurons have been reported to be reduced in localized in neurons, differentially influences post-mortem brains of AD patients. However, brain fatty acid uptake, facilitating the uptake the underlying cause of this reduction is not and trafficking of arachidonic acid (ARA). Our known. Hypothesis and objective: The present observation in brain is a bit different from what study tested the hypothesis that pro-repair lipid we observed for heart fatty acid uptake, mediators are reduced in post-mortem brains of suggesting greater selectivity in the brain. We AD patients because they become trapped have an extensive body of work on the role of within phospholipid and neutral lipid pools via alpha-synuclein (Snca), a non-traditional fatty

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acid binding protein that also binds other lipids, profound reduction of acylcarnitine and myelin in brain fatty acid uptake and metabolism. lipids in a gradient from sciatic nerve, then While Snca influence ARA uptake, it does to by spinal cord, to brain cortex, occurred as early as modulating its metabolism by long chain acyl- one month of age when the blood glucose level CoA synthetases. Further, this leads to an of mice is still normal. We have expanded the important role in brain prostaglandins and 2- findings from lipidomics studies on db/db mice arachidonyl glycerol (2-AG) production. Our to other T2DM models, such as ob/ob and high recent work, in collaboration with Dr. Fred fat diet-induced mice, as well as post-mortem Schroeder, demonstrates that liver FABP brain tissues of T2DM patients. Our mechanistic (FABP1) also has a role in brain ARA levels. In and functional studies have further Fabp1-/- mice, the level of free and total ARA is demonstrated that abnormal nerve lipid elevated in the gene-ablated male mice. This metabolism present in T2DM mice and human increase is similar to the increase observed in tissue is associated with hyperinsulinemia, but the serum of these mice and the levels of the independently of hyperglycemia as evidenced major brain endocannabinoid, 2-AG is increased with the results obtained from streptozotocin- over 2-fold. This is observation is important as treated mice (a T1DM model), and precedes the T94A mutation in FABP1 disrupts its lipid nerve structure/function damage. Taken binding in the liver, suggesting a potential to together, our results challenge the current modulate brain ARA level. Further, this is the hypothesis that diabetic neuropathy is caused first observation that a non-brain localized FABP by long-term hyperglycemia placing influences brain fatty acids levels, suggesting hyperinsulinemia as a novel driver of nerve that there may be a poorly understood damage, and have important implications for interplay between liver lipid metabolism and the diagnosis and treatment of neuropathy in brain fatty acid uptake and metabolism. T2DM.

Shotgun Lipidomics Sheds Light on Diabetic Cytochrome c is a Plasmalogenase that Neuropathy Xianlin Han*, University of Texas Oxidatively Cleaves the sn-1 vinyl Ether Health Science Center at San Antonio, USA Linkage of Plasmalogens Christopher M. Diabetic neuropathy is the most common Jenkins*1, Kui Yang2, Gaoyuan Liu3, Sung Ho diabetic complication and the greatest source Moon3, Beverly G. Dilthey3, and Richard W. of morbidity and mortality in diabetes. The Gross3,1Washington University School of molecular mechanisms underlying diabetic Medicine, USA; 2U.S. Food and Drug neuropathy of type 2 diabetes mellitus (T2DM) Administration, United States; 3Washington are still not completely understood although University School of Medicine, United States hyperglycemia largely attributes to T1DM Plasmalogens are phospholipids critical for diabetic neuropathy. In order to uncover the cell function and signaling that contain a vinyl molecular mechanism(s) leading to T2DM ether linkage at the sn-1 position and are highly neuropathy, we have recently determined the enriched in arachidonic acid (AA) at the sn-2 changes of lipids in brain and nerve tissue position. However, the enzyme(s) responsible samples from db/db mice in a spatiotemporal for the cleavage of the plasmalogen vinyl ether manner by using our enabling shotgun linkage have remained elusive. Based upon the lipidomics technology platform. We have susceptibility of the vinyl ether bond to revealed substantial accumulation of oxidation, we hypothesized that the peroxidase triglyceride and non-esterified fatty acids, and activity of cardiolipin (CL)-activated cytochrome

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c could oxidatively cleave the plasmalogen vinyl Biology, University of Oklahoma Health Sciences ether linkage. Accordingly, we found that Center, Oklahoma City, OK 73104, USA, 5Harold cytochrome c, in the presence of CL, O2 and H2 Hamm Diabetes Center, University of Oklahoma O2 , or oxidized CL and O2 , catalyzed the Health Sciences Center, Oklahoma City, OK oxidation of the plasmalogen vinyl ether bond, 73104, 6Reynolds Oklahoma Center on Aging, promoting its hydrolytic cleavage and the Department of Geriatric Medicine, University of resultant production of 2-AA-lysolipids and α- Oklahoma Health Sciences Center, Oklahoma hydroxy fatty aldehydes. Using stable isotope City, OK 73104 labeling in synergy with strategic chemical Purpose: ELOngation of Very Long chain derivatizations and high-mass-accuracy MS, we fatty acids-4 (ELOVL4) is an elongase deduced the chemical mechanism of this responsible for biosynthesis of very long chain 18 reaction. Specifically, labeling with either O2 (VLC; ≥C28) fatty acids; it makes VLC 18 or H2 O resulted in M + 2 isotopologues of the polyunsaturated fatty acids (VLC-PUFA) in retina α-hydroxyaldehyde, whereas reactions with and testes, and VLC saturated fatty acids (VLC- 18 18 both O2 and H2 O identified the M + 4 SFA) in skin and brain. A 2011 case study linked isotopologue. Furthermore, incorporation of 18O homozygous inheritance of the Stargardt’s 18 from O2 was predominantly located at the α- (STGD3) mutation in ELOVL4 with a central 18 carbon. In contrast, reactions with H2 O nervous system (CNS) phenotype in humans, yielded 18O linked to the aldehyde carbon. including seizures and death. We demonstrate Importantly, cytochrome c released from that ELOVL4-synthesized VLC-SFA play an myocardial mitochondria subjected to oxidative essential regulatory role in pre-synaptic stress cleaved plasmenylcholine in membrane transmission. Methods: We generated an bilayers, and this reaction was blocked by a animal model for STGD3/STGD3 inheritance specific monoclonal antibody against (mut/mut). ELOVL4 localization within the CNS cytochrome c. Collectively, these results identify was determined in wild type mice (wt/wt) using the first plasmalogenase in biology, reveal the immunofluorescence (IF). Synaptic membrane production of signaling lipids by cytochrome c, fractionation was performed on baboon and present new perspectives on cellular hippocampus for lipid analysis. Hippocampal signaling during oxidative stress. slices from (wt/wt) and (mut/mut) were recorded using a multi-electrode array (MEA). Homozygous Expression of Mutant ELOVL4 Primary neuronal cultures from hippocampus of Leads to Seizures and Death in a Novel Animal (wt/wt) and (mut/mut) mice were subjected to Model of Very Long-Chain Fatty Acid FM1-43 assessment of synaptic vesicle Deficiency Blake R. Hopiavuori1,2, Ferenc exocytosis rates in the presence or absence of Deák1,5,6, Richard S. Brush2,3, David M. Sherry1,4, VLC-SFA supplementation. Results: Our Robert E. Anderson*1,2,3,4,6, Martin-Paul STGD3/STGD3 mice recapitulate the human Agbaga1,2,3,4,5, 1Oklahoma Center for phenotype, developing seizures at P19 followed Neurosciences, University of Oklahoma Health by death at P21. Membrane fractionation of Sciences Center, Oklahoma City, OK 73104, baboon hippocampus revealed enrichment of USA, 2Dean McGee Eye Institute, University of 28:0/30:0 in synaptic vesicle membranes. MEA Oklahoma Health Sciences Center, Oklahoma recordings showed a significant dysregulation of City, OK 73104, 3Department of Ophthalmology, both spontaneous and evoked responses in University of Oklahoma Health Sciences Center, (mut/mut) vs (wt/wt) hippocampal slices. FM1- Oklahoma City, OK 73104, 4Department of Cell 43 studies showed a significant increase in

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synaptic vesicle exocytosis rates in (mut/mut) vs causing a CNS phenotype in an animal model. (wt/wt) primary hippocampal neurons and The described studies suggest a neuron-specific supplementation of 28:0 + 30:0 VLC-SFA to the role for VLC-SFA in the regulation of pre- medium rescued release rates back to wild-type synaptic synaptic function by impacting the rate levels, while the LC-SFA, 24:0, did not have a of synaptic vesicle release. significant effect. . Conclusions: This is the first study to demonstrate mutations in Elovl4

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H&N 3: Dairy Fatty Acids and Health Chairs: Moises Torres-Gonzalez, National Dairy Council, USA; and Ignacio Vieitez, UdelaR, Uruguay

Saturated Fat and Cardiometabolic Health cardiometabolic disease risk, based on both Moises Torres-Gonzalez*, National Dairy observational studies and randomized Council, USA controlled trials. Methods Used: Using literature Saturated fat has historically been searches on PubMed and Google Scholar, we discouraged by health authorities because its identified observational studies reporting LDL-cholesterol (LDL-C) rising effects. It is still associations between cardiometabolic disease thought that elevated LDL-C levels increase the endpoints (type 2 diabetes, cardiovascular risk of cardiovascular disease (CVD). However, disease, coronary heart disease, stroke) and (a) recent scientific evidence indicates that LDL-C is both full-fat and low-fat dairy products, and (b) just one of many mediators of the dairy fat intake; and randomized controlled atherosclerosis process and therefore, the trials investigating the effect of interventions prediction of CVD risk is not necessarily rich in full-fat dairy or dairy fat on glucose reflected by the measurements of blood LDL-C homeostasis or cardiovascular disease risk levels. In this sense, emerging scientific factors. Results: Observational studies assessing evidence that indicates that saturated fat the relationship between both low-fat and full- consumption may not be associated with fat dairy food consumption and incident type 2 increased risk of CVD, suggesting that the diabetes or cardiovascular disease are widely relationship between saturated fat, LDL-C and inconsistent. Plasma phospholipid biomarkers CVD is more complex than once thought. The of dairy fat intake are consistently inversely term “saturated fat” has been used to describe associated with incident type 2 diabetes, but mainly the major saturated fatty acids (SFAs) in not with cardiovascular disease. Randomized our diet, e.g. palmitic (C16:0) and stearic (C18:0) controlled trials testing the effect of full-fat acids, however, it results inaccurate because dairy on glucose homeostasis are scarce and oversimplifies the diversity of saturated fatty inconsistent. Intervention trials assessing acids and may mislead consumers to avoid effects of full-fat dairy intake on cardiovascular foods containing “saturated fat” that otherwise disease risk factors show complex effects on the may be more beneficial than detrimental. Thus, serum lipid profile that are hard to interpret, saturated fat is not a single compound but a and that likely depend on the food matrix in family of diverse fatty acids with different which dairy fat is consumed. Conclusions: physiologic, metabolic and health-promoting Overall, the existing knowledge does not properties. During this presentation, a summary suggest consistent negative effects of dairy fat of the emerging scientific evidence on saturated or full-fat dairy product consumption on fat and its relationship with cardiometabolic cardiometabolic disease risk. health will be presented. Effects of Medium- and Long-chain Dairy Fat and Cardiometabolic Health Mario Triacylglycerols on Lipid Metabolism and Host Kratz*, Fred Hutchinson Cancer Research Faecal Microbiota Composition in C57BL/6J Center, USA Mice Shengmin Zhou*1, Yuan Rong Jiang2, and Objective: In this presentation, we will Liangli Yu3,1Wilmar Global R&D Center, China; provide an overview of the current state of 2Wilmar Biotechnology R&D Center (Shanghai) knowledge about the effect of dairy fat on Co., Ltd., China; 3Dept. of Nutrition and Food

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Science, University of Maryland, USA human diet, comprising hundreds of different Objective: In this study, the effects of fatty acids and fatty acid derivatives that fulfill medium- and long-chain triacylglycerols (MLCT) structural, metabolic, and functional roles on lipid metabolism and fecal gut microbiota within the human body. Moreover, the fatty composition of C57BL/6J mice were acid composition of dairy fat is distinct because investigated. Methods: Three MLCT with approximately 70% of the total fatty acids are different contents of medium-chain fatty acids saturated, which arise from the effects of (MCFA) (10%-30%, w/w) were prepared, and ruminal biohydrogenation of dietary fed to C57BL/6J mice. Comprehensive analysis unsaturated fatty acids. Over the past two was used to evaluate the effects of MLCT on the decades, most scientific and public attention lipid metabolism and gut microbiota has focused on this fatty acid class. Yet, dairy fat composition in C57BL/6J mice. Results: MLCT contains a large variety of unique minor fatty with 30% (w/w) MCFA showed the best acid classes or individual fatty acids that are performance in decreasing body weight gain as generated by rumen microbes and the well as optimizing serum lipid parameters and mammary gland. These fatty acids consist of liver triacylglycerol content. The expression short- and medium-chain fatty acids, odd- and levels of genes encoding enzymes for fatty acid branched-chain fatty acids, positional and degradation increased markedly and expression geometric isomers of octadecenoate, and levels of genes encoding enzymes for de novo conjugated linoleic acids. Short-and medium- fatty acid biosynthesis decreased significantly in chain fatty acids are de novo synthesized in the the liver of mice treated with MLCT containing mammary gland utilizing the rumen microbe- 30% (w/w) MCFA. Gut microbiota composition derived fermentation products acetate and showed that the dietary intake of a high fat diet beta-hydroxybutyrate. Branched-chain fatty containing MLCT did significantly decrease the acids arise directly from rumen bacteria via de ratio of Firmicutes to Bacteroidetes and down- novo lipogenesis, whereas isomers of regulate the relative abundance of octadecenoate and conjugated linoleic acids Proteobacteria. In addition, the intake of MLCT originate from incomplete microbial can effectively down-regulate the abundance of biohydrogenation processes of feed-derived Helicobacter. Furthermore, a notable increase polyunsaturated fatty acids in the rumen and was found in the total short-chain fatty acid desaturase activities in the mammary gland. content in feces of mice on a MLCT containing Recent research has indicated that these unique diet. Conclusions: This study shows that MLCT dairy-derived fatty acids possess unusual not only have a direct effect on lipid biological properties and health benefits metabolism by influencing gene expression and beyond their basic nutritional value. The decreasing triacylglycerol content, but also have presentation will summarize the current an effect on gut microbiota composition, which knowledge in the research of dairy-derived fatty may be responsible for the anti-obesity effect of acids with emphasis on potential health MLCT with relatively high contents of MCFA. benefits.

Dairy Fatty Acids: Potential Benefits Jana Other Dairy Fat Components with Potential Kraft*, and Allison Unger, University of Health Benefits: Milk Phospholipids Vermont, USA Christopher N. Blesso*, University of Dairy fat is undeniably the most Connecticut, USA complex and diverse dietary fat source in the Milk fat is encased in a tri-layer milk fat

– 13 – ABSTRACTS 2019 AOCS ANNUAL MEETING & EXPO May 5–8, 2019 globule membrane (MFGM). Phospholipids (C15:0) and Heptadecanoic (C17:0) Odd-chain (PLs), like sphingomyelin (SM), are important Fatty Acids Eneko Ganuza*, and Magdalena components of MFGMs, accounting for about Amezquita, Heliae Development LLC, USA 1% of total milk lipids. The surfactant properties Plasma levels of pentadecanoic (C15:0) of PLs are important for dairy products; and heptadecanoic acid (C17:0) were associated however, dairy products vary considerably in with lower risk of diabetes type-2 and their PL to total lipid content due to processing. cardiovascular disease. Dairy consumption, This presentation will focus on the potential of which is the main dietary source of odd-chain milk PLs in protecting against dysfunctional lipid fatty acid (OCFAs), was also associated with metabolism, inflammation, and gut dysbiosis. lower risks of cardiovascular disease in the Dietary SM has been reported to dose- prospective cohort study “PURE”. These results dependently reduce the intestinal absorption of contrast with the prevailing dietary guidelines cholesterol, triglyceride, and fatty acids in cell against saturated fat intake because roughly 45 culture and rodent studies. Feeding milk PLs has % of it comes from dairy. Milk is predominantly been shown to attenuate some aspects of composed even-chain saturated fatty acids Western diet-induced dyslipidemia, fatty liver, (~70% total fatty acids, TFA) and contains trace gut dysbiosis, and inflammation in rodent amounts of C15:0 and C17:0 (Aurantiochytrium studies. Additionally, feeding milk PLs acetophilum strain-HS399, that is capable of attenuates Western diet-induced accumulating lipids of up to 85% of its weight hyperlipidemia and atherosclerosis in LDL- when grown under heterotrophic conditions. receptor knockout mice. These hypolipidemic This strain naturally produces palmitic (C16:0) effects of milk PLs observed in rodents are also (45%) and docosahexaenoic acid (DHA, C22:6 n- seen in some human studies, although the 3) (40% TFA), but we have modified our extent of reductions in serum cholesterol is fermentation process to synthesize C15:0 typically smaller. This may be explained by the (40%), C16:0 (15%), C17:0 (8%) and DHA (28% more effective digestion of SM in humans and TFA). Hence, we now have the capacity to due to most clinical studies being conducted in produce, at industrial scale, a new nutritional oil healthy populations. Little is known about the that will help us for the first time to impact of milk PLs on inflammation markers and experimentally isolate the effect of dietary odd- the gut microbiota of humans. Milk PLs should chain from even-chain saturated fatty acids. To be considered as food matrix factors that may our knowledge, this is the only natural oil confer health benefits and/or impact effects of containing high concentrations of OCFAs (>49% other dietary lipids, with implications for full-fat TFA), a product that can be used to expand vs. reduced-fat dairy. research on anaplerotic metabolism and biomarkers in epidemiology, and ultimately A Novel Dietary Oil Rich in Pentadecanoic become an important nutritional supplement.

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H&N 4: Health and Nutrition Awards and General Topics Chairs: Matthew Picklo, USDA, ARS, Grand Forks Human Nutrition Research Center, USA; and Elisa Di Stefano, University of Ottawa, Canada

Oxidized dietary fat: a novel risk factor of unknown. A better understanding of the inflammatory bowel disease and colon cancer pathophysiological components involved in IBD via altering gut microbiota Guodong Zhang*, could help to develop novel strategies to reduce UMass-Amherst, USA its risk. Methods and Results: We show that Vegetable oils, such as corn, soybean, systemic treatment with low-dose 4-HNE safflower, and canola oils, consist a substantial exacerbates dextran sulfate sodium (DSS)- part of our diet. Rich in polyunsaturated fats, induced IBD in C57BL/6 mice, suggesting its pro- the vegetable oils are highly prone to oxidation IBD actions in vivo. Treatment with 4-HNE and these oxidation products are commonly suppressed colonic expressions of tight junction found in the diet. However, the effects of protein occludin, impaired intestinal barrier oxidized oils on human health are not well function, and enhanced translocation of understood. Here we show that dietary lipopolysaccharide (LPS) and bacterial products administration of oxidized corn or soybean oil, from the gut into systemic circulation, leading even with low concentrations of lipid oxidation to increased activation of Toll-like receptor 4 products (within the recommended industrial (TLR4) signaling in vivo. Furthermore, 4-HNE limit of oxidative status of fresh vegetable oil), failed to promote DSS-induced IBD in Tlr4-/- exaggerates dextran sodium sulfate (DSS)- mice, supporting that TLR4 signaling contributes induced colitis and exacerbates azoxymethane to the pro-IBD effects of 4-HNE. Conclusions:4- (AOM)/DSS-induced colon tumorigenesis in HNE exacerbate progression of IBD through mice. Oxidized oil reduces the diversity and activation of TLR4 signaling and therefore could alters the composition of gut microbiome, and contribute to the pathogenesis of IBD. fails to promote colitis in antibiotic cocktail- treated mice. These results suggest that Anti-inflammatory Effect of a Novel Metabolite oxidized vegetable oil, even at low oxidative from Marine Carotenoid Fucoxanthin Masashi status, could be a novel risk factor of Hosokawa*, Naoki Takatani, Daisuke Taya, inflammatory bowel disease (IBD) and IBD- Fumiaki Beppu, and Kazuo Miyashita, Hokkaido associated colon cancer. University, Japan Fucoxanthin is a major marine carotenoid 4-HNE, an Endogenous Lipid Peroxidation containing in edible brown seaweeds. It has Product, Exacerbates Colonic Inflammation unique structures such as allenic bond and through Activation of Toll-like Receptor 4 epoxy group in the molecule. We previously Signaling Yuxin Wang*1, Weicang Wang2, and reported that dietary fucoxanthin exhibited Guodong Zhang3,1UMASS, USA; 2UMass- anti-obesity and anti-diabetic effects through Amherst, USA; 3UMass-Amherst, USA prevention of chronic inflammation via Objective: Human and animal studies have adipokine regulation in white adipose tissue shown that the colonic concentrations of lipid (WAT) in diabetic/obese mice. Dietary peroxidation products, such as 4- fucoxanthin is known to be metabolized to hydroxynonenal (4-HNE), are elevated in fucoxanthinol and amarouciaxanthin A in the inflammatory bowel disease (IBD). However, body. Furthermore, we identified an apo- the actions and mechanisms of these carotenoid paracentrone as a novel metabolite compounds on the development of IBD are of fucoxanthin in the mice. Paracentrone

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significantly down-regurated interleukin-6 and hours. Changes in the abundances of cellular monocyte chemoattractant protein-1 mRNA metabolites were identified by high-resolution expressions in macrophage-like RAW264.7 cells MS and MS/MS fragmentation by applying non- activated by LPS. Other apo-fucoxanthinoids targeted metabolomics. In moderately and such as apo-10’-fucoxanthinal also down- highly oxidized grape seed oil, epoxidized and regulated mRNA expression of proinflammatory hydroperoxidized triacylglycerols, such as cytokines in activated RAW264.7 cells. These TG(54:4)OOH, TG(54:5)OOH, TG(54:6)OOH, results suggest that apo-fucoxanthinoids TG(54:0)O, TG(54:1)O, TG(54:2)O, TG(54:3)O, derived from dietary fucoxanthin have anti- TG(54:4)O, TG(54:5)O could be identified, inflammatory effect. whereas the abundances of the epoxidized lipids were markedly diminished in in-vitro Oxidized Triacylglycerols from Grape Seed Oil digested oil samples. Incubation of HGT-1 cells Modulate Phospholipid Pool in Gastric Cells with oxidized triacylglycerols from non- Sarah Frühwirth1, Sofie Zehetner1, Mohammed digested, oxidized grape seed oil induced the Salim1, Sonja Sterneder1, Barbara Lieder1, Luca formation of phospholipids, such as PC(18:2), Nicolotti2, Martin Zehl3, Markus Herderich2, PC(40:7) and PI(20:4). Polar lipids from in-vitro Veronika Somoza1, and Marc digested oil did not affect the phospholipid Pignitter*1,1Department of Physiological metabolism. In-vitro digestion-induced Chemistry, Faculty of Chemistry, University of decrease of epoxidized triacylglycerols might be Vienna, Austria; 2Metabolomics South Australia, responsible for the loss of the effect of dietary Australian Wine Research Institute, Adelaide, oxidized lipids on intracellular phospholipid Australia; 3Department of Analytical Chemistry, pool. Faculty of Chemistry, University of Vienna, Austria Comparison of Postprandial Triacylglycerols Consumption of vegetable oils rich in and Satiety Ratings by Healthy Men Following polyunsaturated fatty acids is considered Ingestion of Tempered Palm Stearin-based healthy. However, these oils are characterized Emulsions Containing Droplets in Different by a lack of oxidative stability, leading to the Physical States Samar Hamad*1, Surangi KPH accumulation of lipid oxidation products, which Thilakarathna2, Amanda Cuncins3, Melissa are supposed to have detrimental effects on Brown2, and Amanda Wright2,1University of health. Studies investigating the metabolic Gueloh, Canada; 2University of Guelph, Canada; response of gastric cells exposed to oxidized 3University of Guelphph, Canada lipids are scarce. We aimed at identifying the With the prevalence of obesity and the effects of oxidized triacylglycerols from relationship between postprandial metabolism differently oxidized and in-vitro digested grape and chronic disease risk, there is growing need seed oils on the cellular metabolism in human to better understand how dietary lipid gastric tumor cells (HGT-1). Moderately properties, including physical state, relate to (peroxide value: 18 meq O2/kg) and highly their digestion and ability to impact satiety. (peroxide value: 28 meq O2/kg) oxidized grape Emulsions containing solid fat droplets versus seed oil was in-vitro digested and analyzed for compositionally equivalent undercooled liquid their antioxidant constituents as well as droplets experienced acid-induced partial oxidized triacylglycerol profile by LC-MS/MS. coalescence and resisted lipolysis under in-vitro HGT-1 cells were treated with isolated fractions digestion conditions. To test the hypothesis that containing oxidized triacylglycerols for up to six this would translate into differences in lipemic

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response and satiety inductions, a double- plasmaglogen (PlsCho), due to little blinded randomized crossover study was fragmentation in MS/MS and co-elution with performed. Fifteen fasting healthy male (age = other phospholipids during column 27.5 ± 5.7 y; BMI = 24.1 ± 2.5; fasting chromatography separations. Recently, we triacylglycerols (TAG) = 0.9 mmol/L) non- overcame those limitations by developing a restrained eaters consumed 50 g of palm sensitive and selective method for the stearin in a 500 mL emulsion beverage which quantitative determination of plasmalogen contained droplets tempered in either the molecular species using LC-MS/MS in the partially crystalline (i.e. SE) or undercooled (i.e. presence of sodium ion. Methods: In this study, LE) state, one at each of two visits. Throughout we quantified plasmalogens including PlsCho the 6-hour postprandial period, participants and ethanolamine (PlsEtn) species, in serum provided blood samples and completed visual (n=25-29 per group) and postmortem pre- analogue satiety scales. Preliminary results frontal cortex (n=20-21 per group) of AD reveal that LE had a higher AUC lipemic patients and age-matched healthy controls. We response and earlier rise from baseline hypothesized that peripheral and central compared to SE. Change from baseline values disturbances in plasmalogen species would be for Hunger, Appetite and overall Average observed in AD patients. Results: In serum, Appetite were consistently lower, and Fullness PlsCho and PlsEtn species bearing was consistently higher, for LE compared to SE, docosahexaenoic acid (DHA) were significantly but no significant differences were observed increased in AD patients compared to controls. (P>0.05). Therefore, emulsion droplet solid fat In post-mortem AD brains, PlsCho bearing DHA was associated with small differences in and PlsEtn bearing arachidonic acid (ARA) were postprandial TAG, although these did not reduced, whereas alkyl phosphatidylcholine correspond with differences in participant bearing DHA was increased. The results suggest ratings of satiety. dysregulated metabolism of DHA and ARA bearing plasmalogens in serum and brain of AD Analysis of Plasmalogen Species in Serum and patients. Post-mortem Brain Tissue of Alzheimer’s Disease Patients Yurika Otoki*1, Shunji Kato1, Applying Carbon-13 Natural Abundance in Kiyotaka Nakagawa1, Di Yu2, Danielle J. Harvey3, Human Plasma as a Tool for the Assessment of Lee-Way Jin3, Britany N. Dugger3, Walter n-3 Polyunsaturated Fatty Acid Metabolism Swardfager2, and Ameer Taha4,1Tohoku Adam H. Metherel*1, Maha Irfan1, Shannon L. University, Japan; 2Sunnybrook Research Klingel2, David M. Mutch2, and Richard P. Institute, Canada; 3University of California, Bazinet1,1University of Toronto, Canada; Davis, USA; 4University of California, Davis, USA 2University of Guelph, Canada Background and hypothesis: Alzheimer’s Carbon-13 isotopic abundance (δ13C) of disease (AD) is a progressive brain disorder tissue and blood n-3 polyunsaturated fatty acids characterized by the deposition of abnormal (PUFA) can answer complex metabolic protein aggregates and loss of neurons. In AD questions in rodent models; however, to date, brains and blood, marked changes in ether- these methods have not been extensively linked phospholipids known as plasmalogens, applied in humans. Our objective was to track have been reported. The complement of the changes in δ13C in human plasma following molecular plasmalogen species have been eicosapentaenoic acid (EPA) or difficult to measure, especially choline docosahexaenoic acid (DHA) supplementation.

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Participants (aged 21.6 ± 2.2 y) were randomly approach, ~100 mg of colon tissues from 8 mice allocated into one of three supplement groups per group were mixed with an antioxidant for 12 weeks: 1) olive oil control, 2) ~3 g/d EPA solution (0.2 mg/mL butylated hydroxytoluene or 3) ~3 g/d DHA. Blood was collected before and 0.2 mg/mL triphenylphosphine in and after the supplementation period, and methanol), 10 µL of deuterated internal plasma fatty acid concentrations and δ13C of n-3 standards (500 nM of d4-6-keto PGF1a, d4- PUFA were determined. EPA supplementation TXB2, d4-PGE2, d4-LTB4, d11-14,15-DHET, d4-9- increased (p < 0.05) plasma docosapentaenoic HODE, d8-5-HETE, d11-11,12-EET), and 400 μL acid (DPAn-3) concentrations by 9-fold and 2- extract solution (0.1% acetic acid with 0.2 fold, respectively, but did not affect DHA mg/mL butylated hydroxytoluene in methanol), concentrations. However, both plasma δ13C- and then homogenized. The lipid metabolites in DPAn-3 (-28.9 ± 0.3 to -25.0 ± 0.1, milliUrey ± the combined solutions were loaded onto pre- SEM) and δ13C-DHA increased (-27.9 ± 0.2 to - washed Waters® Oasis solid phase extraction 25.6 ± 0.1, p < 0.05) to values close to δ13C-EPA HLB cartridges, washed with 95:5 v/v in the supplement (-23.5 ± 0.22). DHA water/methanol with 0.1% acetic acid, the supplementation increased (p < 0.05) plasma analytes were eluted with methanol and ethyl EPA by 130%, but plasma δ13C-EPA was acetate, dried using a centrifugal vacuum unaffected by supplementation. Our study is evaporator, then reconstituted in methanol for the first to assess changes in human δ13C-PUFA LC-MS/MS analysis. The LC-MS/MS analysis was following either EPA or DHA supplementation. carried out on an Agilent 1200SL HPLC system We provide clear evidence for significant (Agilent, Santa Clara, CA) coupled to a 4000 metabolic conversion of EPA to DHA with EPA QTRAP MS/MS (AB Sciex, Foster City, CA). The supplementation, and the absence of peaks were identified according to the retroconversion of DHA to EPA with DHA retention time and specific multiple reaction supplementation. δ13C of plasma n-3 PUFA in monitoring transitions of the standards of lipid humans is a powerful tool that can track dietary metabolite. The concentrations of the lipid n-3 PUFA to elucidate complex metabolic metabolites were calculated against the questions. calibration curve with standards. To study the functional role of sEH in obesity-induced colonic Lipidomic Profiling Reveals Soluble Epoxide inflammation, we treated mice with sEH Hydrolase as a Therapeutic Target of Obesity- inhibitors, TPPU or t-TUCB, in a high-fat diet induced Colonic Inflammation Weicang (HFD)-induce obesity model. Result: Using an Wang*1, Jun Yang, Jianan Zhang , Yuxin Wang1, LC-MS/MS-based lipidomics approach, we Katherine Z. Sanidad2, Zhenhua Liu2, Bruce D. found that obesity-induced colonic Hammock, and Guodong Zhang3,1UMass- inflammation is associated with increased Amherst, USA; 2University of Massachusetts- expression of soluble epoxide hydrolase (sEH) Amherst, USA; 3UMass-Amherst, USA and its eicosanoid metabolites, termed fatty Background: Obesity is associated with acid diols, in colon tissue. Pharmacological enhanced colonic inflammation, which is a inhibition of sEH attenuated obesity-induced major risk factor of colorectal cancer. colonic inflammation by diminishing the Considering the obesity epidemic in the United infiltration of inflammatory cells and decreasing States, there is an urgent need to identify novel the gene expressions of the pro-inflammatory therapeutic targets for obesity and its related cytokines in colon tissues in obese mice. disease. Method: In LC-MS/MS-based lipidomics Furthermore, we examined the sEH genetic

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knockout mice in obesity model, and found caloric intake was far more important than fat genetic ablation of sEH reduced colonic intake for promotion of tumor growth around concentrations of fatty acid diols and decreased the time the low-fat message became widely the gene expressions of pro-inflammatory accepted. Both colon and breast cancers were cytokines. Finally, HFD treatment increased inhibited by caloric restriction (CR) no matter expression of phosphorylated GSK3β in colon how much fat was fed. Other labs tissue, while such effect was attenuated in sEH demonstrated similar effects of CR in a variety genetic knockout mice, or by treatment with of tumor models. We went on to determine sEH inhibitors, suggesting that pharmacological that approximately 25% CR was required to inhibition and genetic ablation of sEH significantly inhibit tumor promotion, that attenuated HFD-induced activation of Wnt genetic obesity markedly promoted tumor signaling in the colon. Conclusion: Together, growth, and that body fat was also not a these results support that sEH could be a novel significant factor in tumor promotion. These therapeutic target for obesity-induced colonic results helped set the stage for the Women’s inflammation and associated diseases. Health Initiative that tested low-fat diet against Moreover, our study would lead to rapid human the average American diet in 50,000 people for translation, as pharmacological inhibitors of sEH reductions in multiple chronic diseases and are being evaluated in human clinical trials found no significant benefit on any primary targeting multiple disorders. endpoint. Collectively, animal studies and human diet interventions did not support the Why Fat Was the Dietary Devil … and Then it observational associations of high dietary fat Wasn’t David M. Klurfeld*, USDA Agricultural and cancer. The same is true today for claims Research Service, USA about meat and cancer. This example of Epidemiologic and animal studies experimental data as a catalyst for overturning beginning in the 1960’s suggested that dietary accepted dogma in nutrition demonstrates that fat increased the risk of several types of cancer. the field must embrace higher standards for For the last four decades, the number one reaching conclusions about causality. dietary advice was to eat less fat. Our lab began publishing in the 1980’s a series of studies that

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H&N-P: Health and Nutrition Poster Session Chair: Matthew Picklo, USDA, ARS, Grand Forks Human Nutrition Research Center, USA

1. Effect of Whey Peptides on Metabolism peptides promote differentiation and PPARγ and Insulin Signaling in Muscle and Fat Cells. activation in adipocytes. In myotubes exposed Kenneth D'Souza*1, Angella Mercer1, Hannah to an obese-diabetic milieu, whey peptides Mawhinney2, Thomas Pulinilkunnil1, Chibuike C. ameliorate insulin resistance, potentially by Udenigwe2, and Petra Kienesberger1, 1Dalhousie reducing inflammation and ER stress. Taken Medicine New Brunswick, Canada; 2University of together, our data suggest that whey peptides Ottawa, Canada directly enhance lipid metabolism and insulin RATIONALE: Adipose tissue and skeletal function in adipocytes and muscle cells. muscle dysfunction are hallmarks of obesity and insulin resistance. Bioactive peptides derived 2. Ursolic Acid Attenuates 6- from food sources including milk and dairy Hydroxydopamine Induced-Apoptosis through products have gained interest for their roles in Up-Regulation of OPA1 in SH-SY5Y Cells. Fang- obesity and insulin resistance. However, it Ting Chang, Chia-Yuan Lin, and Chia-Wen Tsai*, remains unclear whether and how whey China Medical University, Taiwan impacts adipose and muscle metabolism and Ursolic acid attenuates 6-hydroxydopamine insulin function. HYPOTHESIS: Bioactive whey induced-apoptosis through up- regulation of peptides have an insulin-sensitizing effect on OPA1 in SH-SY5Y cells Fang-Ting Chang, Chia- adipocytes and muscle cells. METHODS: Whey Yuan Lin and Chia-Wen Tsai* Department of peptide mixture was generated via the Nutrition, China Medical University, Taiwan hydrolysis of whey protein with pepsin and *[email protected] Hypothesis: pancreatin. 3T3-L1 pre-adipocytes were Mitochondrial dynamic imbalance is related to incubated with 2.5 mg/ml bovine serum the pathogenesis of Parkinson’s disease (PD). albumin (BSA) or whey peptides during Optic atrophy 1 (OPA1), a mitochondrial fusion differentiation. Lipid accumulation and the protein, is responsible for maintenance of expression of proteins involved in lipid mitochondrial integrity and protection from cell metabolism was analyzed. Insulin resistant death. In this study, we explored whether C2C12 myotubes were incubated with BSA or ursolic acid (UA) obtained from herbs and fruits whey peptides for 16 h followed by insulin could protect SH-SY5Y neuronal cells against 6- signaling analysis. RESULTS: In 3T3-L1 cells, OHDA induced apoptosis by up-regulation of whey peptides increased expression of the OPA1. Methods: Western blotting was used to master-regulators of adipogenesis, C/EBPα and detect the protein expression. The method of PPARγ. This was associated with upregulation of small interfering RNA transfection was used to perilipin and adiponectin, markers of enhanced explore the effect of knockdown of parkin and lipid storage and insulin sensitivity, and OPA1. Immunoprecipitation was used to increased triacylglycerol accumulation. In C2C12 explore whether IκB kinase (IKKγ) interacted myotubes, whey peptides protected from with ubiquitin by UA. Results: Cells pretreated palmitate-induced insulin resistance, as with UA attenuated the cell death, nuclear determined by improved AKT phosphorylation condensation, and cleavage of caspase 3 and Glut4 expression. Insulin sensitization of induced by 6-OHDA. Moreover, UA reversed 6- C2C12 myotubes was accompanied by OHDA-reduced the protein expression of parkin decreased inflammation and ER stress following and OPA1. UA alone increased the parkin whey peptide treatment. CONCLUSIONS: Whey protein, enhanced the interaction of ubiquitin

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and IKKγ, activated the p65 pathway and 0.58%) and docosahexaenoic (C22:6ω3, 0.49 vs induced the OPA1 protein. Suppression of 0.85 or 1.13%) acids in LD muscle. The results parkin reduced the ability of CA on p65 indicate that feeding lambs with the protecting activation and OPA1 induction. In cells-treated dietary supplements prepared with GRAS with OPA1 siRNA, UA could no longer chemicals can increase the deposition of α- significantly reverse the apoptosis and nuclear tocopherols and ω-3 fatty acids in their muscle. condensation by 6-OHDA. Conclusions: UA increases the protein expression of OPA1 4. Identification of Angiotensin-converting through parkin regulated NFκB pathway, Enzyme Inhibitory Peptides Resisting leading to prevent the apoptosis by 6-OHDA. Gastrointestinal Digestion from Thermolysin- digested Egg White Hydrolysate. Hongbing 3. Incorporation of Antioxidants and Omega- Fan*, Jiapei Wang, and Jianping Wu, University 3 Fatty Acids in Fresh Lambs by Feeding of Alberta, Canada Rumen-protected Dietary Supplements. As a high quality protein, the hen’s egg Chelsea Jeanjulien1, Jung Hoon Lee*1, and proteins are recognized as excellent sources of Stephan Wildues2, 1Fort Valley State University, bioactive peptides, such as angiotensin- USA; 2Virginia State University, USA converting enzyme inhibitory (ACEi) peptides. Incorporation of antioxidants and ω-3 fatty Our previous research showed that egg white acids in fresh meat can provide healthier meat hydrolysate (EWH), prepared by thermolysin for human and extend the shelf life of fresh and then pepsin, decreased blood pressure meat. Twenty-four lambs were used to assess significantly in spontaneously hypertensive rats the effects of rumen-protected dietary (SHRs); further investigation showed that supplements (RPSD) containing ω-3 fish oil (140 thermolysin-digested EWH (T-EWH) could also g/kg) and DL-α-tocopheryl acetate (TOA: 759 reduce blood pressure significantly in SHRs. In mg/kg) with or without L-ascorbic acid (ASC: addition, in vitro ACEi assay showed that the 784 mg/kg), prepared with a GRAS chemical on ACEi activity of T-EWH could be enhanced by fresh lambs. Lambs were assigned to one of simulated gastrointestinal (GI) digestion. These three dietary treatments, which consisted of results indicated that T-EWH might contain ACEi corn basal diet (CBD) with TOA and ASC, and peptides with excellent GI stability. Therefore, 86.7% CBD plus either RPSD containing TOA the purpose of this study is to identify ACEi only or TOA and ASC. Lambs were harvested peptides in T-EWH that resist GI digestion. After after a 49-d of feeding trial, and muscle tissues conventionally activity-guided fractionation, all were obtained from each carcass to analyze α- fractions were subjected to simulated GI tocopherol and ascorbic acid contents, and fatty digestion, before and after which their ACEi acid compositions. Compared with lambs fed a activities were both determined. Finally, six new CBD diet, lambs fed both RPDS containing diets ACEi peptides (LAPYK, LKISQ, LKYAT, INKVVR, had higher (P < 0.05) concentrations of α- LFLIKH, and LGHWVY), resisting gastrointestinal tocopherol (LD muscle, 1.15 vs 2.73 and 3.84 digestion, were identified and validated, with μg/g; psoas major, 1.36 vs 3.57 and 4.34 μg/g; IC50 values < 15 uM, respectively. gluteus medius, 1.51 vs 4.18 and 4.50 μg/g) in muscle samples. Compared with lambs fed CBD, 5. Effects of Dietary Scallop Oil Prepared lambs fed either RPDS containing TOA or TOA + from Scallop By-product on Lipid Metabolism ASC had higher (P < 0.05) concentrations of in Type II Diabetic/Obese KK-Ay Mice. Koki eicosapentaenoic (C20:5ω3, 0.24 vs 0.48 or Sugimoto*1, Junpei Nakamura1, Ryota Hosomi1,

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Munehiro Yoshida1, and Kenji Fukunaga2, 6. Dietary n-3 polyunsaturated Fatty Acid 1Kansai University, Japan; 1Faculty of Chemistry, Ethyl Ester Influences the Composition of Materials and Bioengineering, Kansai Bacteria and Their Metabolites in Rat Cecal University, Japan Content. Ryota Hosomi*1, Anna Matsudo1, Internal organs of scallops, such as, the Tadahiro Tsushima2, Yoshihisa Misawa2, Takaki midgut glands are rich in eicosapentaenoic and Shimono3, Seiji Kanda3, Toshimasa Nishiyama3, docosahexaenoic acid. Scallop by-products have Munehiro Yoshida3, and Kenji Fukunaga4, not yet been utilized to their full potential 1Kansai University, Japan; 2Bizen Chemical C. Co. because they have high concentrations of toxic Ltd., Japan; 3Kansai Medical University, Japan; substances including cadmium and arsenic. We 4Faculty of Chemistry, Materials and have successfully prepared scallop oil (SCO) Bioengineering, Kansai University, Japan containing low toxic substances from scallop by- Numerous studies have evaluated the gut products. In this study, we prepared SCO from microbial flora of high-fat or low-fiber diet fed scallop by-products obtained from two different animals under an experimental set-up. On the production areas and referred to them as SCOM other hand, only a few reports have focused on (SCO from Mutsu, Aomori), and SCOU (SCO the effect of dietary fatty acids on bacterial from Uchiura, Hokkaido). The aim of this study flora and their metabolites. Our previous study was to investigate the mechanism by which showed that menhaden oil, a eicosapentaenoic dietary SCO influences lipid metabolism in KK- acid (EPA) rich oil, had a different effect on the Ay mice. Four-week old male KK-Ay mice were composition of bacteria and their metabolites in sub-divided into four groups. Mice in the rat cecal content compared to soybean oil (SO), control group were fed a AIN93G modified high- lard, and tuna oil (2018 AOCS Annual Meeting). fat (3% soybean oil plus 17% lard) diet, and Therefore, the aim of this study was to other groups were fed high-fat diet, where 7% investigate the effect of dietary n-3 of the lard was substituted with SCOM, SCOU, polyunsaturated fatty acid (PUFA) ethyl ester or tuna oil. After 7 weeks, serum triacylglycerol (EE) on the bacterial composition and their levels decreased significantly in the SCOM, metabolites in rat cecal content. Male Wistar SCOU, and tuna oil-fed mice compared to the rats (4-weeks old) were divided four groups and control group. Epididymal adipose tissue weight fed a high-fat diet containing 15% SO, 10% SO + significantly decreased in the SCOU group 5% SO-EE, 10% SO + 5% EPA-EE, or 10% SO + 5% compared to the control group. The hepatic docosahexaenoic acid (DHA)-EE. After 28 days, mRNA levels of fatty acid synthase, acetyl-CoA the bacterial composition of cecal content was carboxylase, and stearoyl-CoA desaturase assessed by sequencing 16S ribosomal RNA tended to decrease while expression of gene, and bacterial metabolites including carnitine palmitoyltransferase-1, peroxisome neutral and acidic steroids were measured. Rats proliferator-activated receptor alpha, and acyl- fed DHA-EE diet had a high relative abundance CoA oxidase tended to increase in the SCOM of Bacteroidetes and low proportion of and SCOU groups, respectively, compared to Firmicutes compared to the rats fed SO-EE and the control group. Thus, dietary SCOM and EPA-EE diet. Fecal secondary bile acids SCOU may ameliorate lifestyle-related diseases significantly decreased in EPA-EE and DHA-EE such as hyperlipidemia and obesity. diet compared to SO-EE diet. Interestingly, the EPA-EE diet significantly decreased the fecal deoxycholic acid and lithocholic acid contents compared to the SO-EE diet. Thus, not only the

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amount of dietary fat but also the fatty acid in HUVECs, due to the abundant presence of composition altered the composition of bacteria CaSR in the cell membrane and cytosol of and their metabolites. HAECs. Thus, the study concluded that γ-EV could significantly reduce vascular inflammation 7. Evaluate the Efficacy of γ-glutamyl Peptide in HAECs, but not in HUVECs, through CaSR in Reducing Hypertension-associated Vascular dependent pathways. Inflammation. Snigdha Guha*1 and Kaustav Majumder2, 1University of Nebraska, Lincoln, 8. Metabolomics Revealed the Selective USA; 2University of Nebraska, USA Effects of Feeding Oxidized Oil on Amino Acid Nearly 54% of the 75 million adults in the Metabolism in Nursery Pigs. Yue Guo*1, Andrea U.S who are suffering from hypertension lack in Hanson1, Lei Wang1, Brian Kerr2, Pedro Urriola1, appropriate preventative intervention to tackle Gerald Shurson1, and Chi Chen1, 1University of the associated risk factors such as heart failure Minnesota, USA; 2USDA-ARS, USA and atherosclerosis. It is well-documented that Oxidized oils from rendering and recycling hypertension develops from a complex are an economic source of energy and lipids in interaction between genetic and environmental swine feeding. Negative effects of feeding factors and increased levels of vascular oxidized oils on pigs have been observed, but inflammation is a significant contributor to the the underlying metabolic events remain to be pathogenesis of hypertension. Dry-edible beans defined. In this study, oxidized corn oil (OCO) derived γ-glutamyl peptide (γ-EV) can reduce was produced by heating control corn oil (CCO) gastrointestinal inflammation via allosteric at 185 oC for 12 h. Weanling pigs were assigned activation of Calcium-Sensing Receptors (CaSR). to 4 isocaloric diets containing 9% CCO, 6% CCO Since vascular cells also express CaSR, it was + 3% OCO, 3% CCO + 6% OCO, and 9% OCO for hypothesized that the intervention of γ- 35 days, respectively. Feeding OCO reduced glutamyl peptide (γ-EV) might modulate gain:feed ratio. Serum and liver samples inflammatory responses in vascular cells via the collected on d 35 were analyzed by the liquid activation of CaSR. To determine the anti- chromatography-mass spectrometry (LC-MS)- inflammatory effects, Human Aortic Endothelial based metabolomics analysis. In the liver, OCO Cells (HAEC) and Human Umbilical Vein decreased the levels of hepatic glutamic acid, Endothelial Cells (HUVEC) were first pre-treated methionine, and glycine, and also the ratio of (2h) with different concentrations of the γ-EV glutathione (GSH) / glutathione disulfide (0.01mM, 0.1mM, and 1 mM), followed by the (GSSG), while increased the gene expression treatment with human TNF-ɑ (5ng/mL, 6h) to levels of GSH metabolism enzymes. Similarly, induce the inflammation. Western blot and OCO decreased the level of tryptophan in serum ELISA were performed to measure the as well as the levels of carnosine in both serum expressions of pro-inflammatory cell adhesion and liver, while increased the gene expression molecules and chemokines respectively. It was levels of the enzymes for tryptophan and observed that TNF-ɑ treatment significantly carnosine catabolism in the liver. In contract, increased the expression of the pro- feeding OCO increased the level of serum inflammatory cell adhesion molecules (ICAM-1 threonine and decreased the levels of threonine and VCAM-1) and chemokine (MCP-1). metabolites in the liver. Overall, these results However, the γ-EV pre-treatment at 1mM suggests that consuming oxidized oils can significantly down-regulated the expression of selectively modify amino acid metabolism of ICAM-1, VCAM-1, and MCP-1 in HAECs, but not pigs, which may negatively affect health and

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growth performance. intervention test, among 18 subjects was control group and others were experimental 9. The Mechanisms of Hypoglycemic Effect of group. Subject was asked to take foxtail millet- Foxtail Millet Based on Transcriptomics. Fu derived products containing 50 g of foxtail Yongxia*, Fan Zhang, Xin Ren, and Qun Shen, millet each day for 90 days. The blood glucose, China Agricultural University, China blood lipid, blood pressure, kidney function, Our previous animal experiment have cytokines and other body indexes were fully proved that foxtail millet feeding significantly investigated at week 0, 6, and 12, respectively. improved the blood glucose metabolism in The intake of 50g of foxtail millet per day after high fat diet and STZ-induced (HFD/STZ) 12 weeks significantly decreased fasting blood diabetic rats. To further clarify the mechanisms glucose compare with the very beginning (5.74 of the hypoglycemic effect of foxtail millet, the ± 0.12 at week 0 vs 5.30 ± 0.09 at week 12), 2h- difference on liver transcriptional profiles glucose after oral glucose tolerance test (OGTT, between diabetic rats, normal rats and foxtail 10.21 ± 0.33 at week 0 vs 9.36 ± 0.28 at week millet feeding rats were investigated. The 12), HOMA-IR (4.16 ± 0.33 at week 0 vs 3.34 ± results shown that 4 weeks of foxtail millet 0.25 at week 12) and obesity degree (110.03 ± feeding in this study could mitigate negative 1.90 at week 0 vs 108.33 ± 1.73at week 12) and variations of liver transcriptional profile in significantly increased leptin concentration HFD/STZ diabetic rats. Specifically, foxtail millet (8.30 ± 0.82at week 0 vs 9.55 ± 0.89 at week 12) feeding activated the insulin-stimulated in subjects with IGT. The intake of 50 g of foxtail PI3K/AKT signaling pathway by up-regulating millet also has trend to lower the blood lipid expression of IRS, PI3K and AKT in liver, which and pressure, and increasing kidney function, will inhibit gluconeogenesis by down-regulating bone density and body inflammation. Foxtail expression of G6P, FBP and PEPCK, and millet diet significantly improved the glycemic stimulate glycolysis by up-regulating expression control, especially the postprandial glucose, by of GK and PK subsequently. Moreover, foxtail increasing insulin sensitivity and leptin millet feeding inhibited NF-κB signaling pathway concentration and alleviating inflammatory and reduced expression of inflammatory condition in subjects with IGT. factors, which will weaken the inhibition of insulin signaling pathway and improve blood 11. Time-Dependent Differences in Essential glucose metabolism ultimately. Keywords: Omega-3 Fatty, during Lactation in the foxtail millet; blood glucose metabolism; insulin Guatemalan Highlands. Doug Bibus1*, Heike signaling pathway; NF-κB signaling pathway Rolker2, Alejandra Zamora2, Rosario Garcia- Meza2, Claudia Arriaga2, and Noel W. 10. Improving Glycemic Control with Foxtail Solomons2, 1Lipid Technologies, LLC and The Millet Foods and How to Get Them in the Center for Spirituality and Healing, University of Marketplace. Fan Zhang*, Fu Yongxia, Ren Xin, Minnesota, USA, 2Center for the Studies of and Qun Shen, China Agricultural University, Sensory Impairment, Aging and Metabolism, China Guatemala The study was to investigate the effect of Objective: To examine time-dependent foxtail millet on glycemic control in subjects differences in the long-chain omega-3 fatty with impaired glucose tolerance (IGT). 82 acids, DHA and EPA, in lactating Guatemalan subjects with IGT were separated into two women. Methods: Full-breast milk extractions groups for a 12-week foxtail millet diet were obtained from 40 women with healthy

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infants. Breast milk was sampled at or near four Minnesota, USA, 2Center for the Studies of time-points including 40 days (44±3); 80 days Sensory Impairment, Aging and Metabolism, (73±4); 120 days (120±7) and 160 days (160±9). Guatemala Four spots of breast milk were applied to a Objective: To assess serial values of DHA stabilized PerkinElmer 226 five-spot RUO spot and EPA in human milk using self-sampling card, dried at room temperature for 60 min and dried milk spots in response to daily then frozen-stored until shipping to Austin, MN, administration of these fatty acids. Methods: USA. Breast milk samples were directly Ten women between 28 and 56 days of esterified with acidic methanol and analyzed by lactation were stratified to receive either capillary gas chromatography, reporting 42 fatty 450mg DHA/90 mg EPA (full dose ‘FD’) or 225 acids, from C-14 to C-24 and expressed as mg DHA/45mg EPA (half dose ‘HD’) as a weight %. We modeled the daily amount of formulated supplement over a 4-week period. DHA and EPA consumed by infants using the Subjects self-collected breast milk samples at assumptions of 780ml of milk consumed per home on a stabilized PerkinElmer 226 RUO spot day with a milk fat content of 34 g/L equally card on an alternate-day sampling schedule: 26.5g of fat per day. Results: The serial amounts baseline; x6 in the first 2 weeks; and x4 in the of DHA (wt%) were: 0.14±0.03 (T40); 0.16±0.07 last 2 weeks for a total of 11 card-based (T80); 0.10±0.03 (T120) and 0.21±0.16 (T160). samples. After frozen-shipment to Austin, MN, By Kruskal-Wallis analysis, the value at T120 breast milk samples were directly esterified was significantly lower than the value at T160 . with acidic methanol and analyzed by capillary Based on the corresponding daily median gas chromatography, reporting 42 fatty acids intakes, DHA(mg) intake models to: 35.9 (T40); from C-14 to C-24. Data are expressed as weight 35.9 (T80); 23.8 (T120) and 35.9 (T160). For %. Results: Average DHA content at baseline EPA, the parallel data are: 0.03±0.01 (T40); was 0.19 +/-0.08 (median 0.19 and range 0.09- 0.04±0.02 (T80); 0.02±0.004 (T120) and 0.34 (n=10)). Corresponding data for EPA was: 0.03±0.03 (T160), corresponding to daily 0.04+/-0.03(median 0.04 and range 0.01-0.10 median intakes in mg of: 7.8 (T40); 10.4(T80); (n=10)). Median change from baseline to end 5.2 (T120) and 5.2 (T160). Conclusions: In the for FD supplementation for DHA was Δ0.35, context of living conditions and lactation in from 0.19 to 0.54 (p=0.0033, n=5), representing Guatemala, there is significant variation in the an increment from 20th to 90th percentiles. abundance of milk DHA and EPA. In comparison With HD, response was Δ0.16, from 0.19 to 0.35 to populational breast milk DHA data, (p=0.019, n=5), to the 65th percentile. The Guatemalan women have lower levels of DHA corresponding median EPA responses were and may benefit from increased dietary DHA. Δ0.06 (p=0.004, n=5) and Δ0.02 (p=0.03, n=5). Conclusion: Empowering Guatemalan women 12. Using Dried Human-milk Spots to with the dried-milk-spot approach allowed Document Time-Dependent Omega-3 Content demonstration of the substantial impact of oral with Oral DHA and EPA Supplementation of EPA and DHA supplementation into upper Guatemalan Mothers. Doug Bibus1*, Rosario percentiles of human milk content in only 4 Garcia-Meza2, Marta Escobar2, Debora weeks. Improved breast milk DHA status may Fuentes2, Alejandra Maldonado2, Alejandra offer improved neural developmental and Zamora2, Claudia Arriaga2, and Noel W. cognitive outcomes in Guatemalan children. Solomons2, 1Lipid Technologies, LLC and The Center for Spirituality and Healing, University of

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13. Extracellular vesicles isolated from two scale to use them as antioxidant delivery types of olive coproducts Jerôme Lecomte1, vesicles. Bruno Baréa2, Marie-Caroline Michalski3, Claire Bourlieu4, Pierre Villeneuve5, Amal Fenaghra*6, 14. Characterization of Phenolic Profile in Pascal Colosetti7, and Anne Mey7,1CIRAD, Wheat Co-Products Determined by UPLC-MS Greece; 2CIRAD, France; 3INRA, Francce; 4UMR Erwann Durand1, Claire Bourlieu2, Véronique IATE - INRA/CIRAD/UM2/SupAgro, France; Vié3, Pierre Villeneuve4, Millena Barros 5UMR IATE, CIRAD, France; 6UMR IATE, France; Santos*5, Cécile Barron5, Valérie Micard5, 7UMR CarMeN, France Valérie Lullien-Pellerin5, L Cameron6, and Extracellular vesicles (EVs) are nanoscopic Mariana Ferreira6,1CIRAD, France; 2UMR IATE - membranous structures which are involved in INRA/CIRAD/UM2/SupAgro, France; 3Institute of intercellular communication between cells. Physics Rennes, University of Rennes 1, France; Recent works highlighted the existence of EVs 4UMR IATE, CIRAD, France; 5UMR IATE, France; secreted by plant cells. The objective of our 6Federal University of State of Rio de Janeiro, study, is to characterize the main physico- Brazil chemical properties of EVs originated from olive Wheat co-products are rich in secondary coproducts (pomace -PEVs- and wastewaters - metabolites that show a wide spectrum of WWEVs-). Isolation was carried out using bioactivities, such as phenolic compounds (PC). successive centrifugations. The main physical PC have a moderate to low solubility in apolar characterizations were size and charge using media that restricts their application in oil- Dynamic Light Scattering (DLS). The chemical based products. The choice of the extraction characterization included phenolic compounds solvent is essential to maximize the yield, characterization using HPLC and lipid classes recovery and subsequent application of PC. with TLC. Furthermore, test of their antioxidant Among green solvents, deep eutectic solvents effect was conducted using CAT assay. EVs (DES) have emerged as effective media to isolated from the two co-products presented extract PC because of their ability to improve distinct physico-chemical properties. WWEVs their solubility, stability and compatibility with showed a larger diameter than PEVs that enzymatic reactions, especially when used as a ranged between 20-140 nm versus 20-50nm. media of lipase functionalization. In this work, On a [2-7] pH range, evolution of charge PC were extracted by three conventional resulting in different curve shapes for the two solvent mixtures (ethanol:water, VEs but indicated similar isoelectric point [2-3]. isopronanol:water and acetone:water) and two WWEVs were more concentrated with phenolic formulations of DES (choline chloride:2- compounds (0.5mM eq HT) and contained more propanediol – DES1 and choline chloride:latic neutral lipids than PEVs. However similar polar acid – DES3) and characterized by UPLC-ESI- lipids classes were found in the two VEs. QTOF-MSE in negative mode. Data were WWEVs was the fraction with the highest processed using Progenesis QI with a antioxidant activity. EVs originated from customized databank of polyphenols applying wastewaters showed a better profile in lipid exact mass error (<10 ppm) and isotope and phenolic profile, but due to environmental similarity (>80%). A total of 75 compounds were considerations, working with pomace may be tentatively identified. The flavonoid apigenin 7- preferable. In perspectives, EVs characteristics O-apiosyl glucoside was the most abundant will be mimicked and improved at laboratory compound, present in all extracts. Wheat bran showed a higher number of PC identifications

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than aleurone extracts. Conventional solvents showed a higher number of PC identifications, probably due to some sort of incompatibility of DES based extracts in MS approaches. However and remarkably, DES3 extracted 9 unique PC, demonstrating that it is a good media to extract specifics wheat PC and to add value to this abundant low-cost by-product of the milling industry.

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