Background Rice biofortification with Zinc (Zn) can improve the Zn status of rice-consuming populations. However, the metabolic impact in humans consuming Zn-biofortified rice is unknown. Objectives To determine the effects of Zn-biofortified rice on lipid metabolism in normolipidemic men. Methods The men consumed a rice-based diet containing 6 mg Zn/d and 1.5 g phytate (phytate/Zn ratio = 44) for 2 wk followed by a 10-mg Zn/d diet without phytate for 4 wk. An ad libitum diet supplemented with 25 mg Zn/d was then fed for 3 wk. Fasting blood samples were taken at baseline and at the end of each metabolic period for measuring plasma zinc, glucose, insulin, triglyceride (TG), LDL and HDL cholesterol, fatty acids, oxylipins, and fatty acid desaturase activities. Statistical differences were assessed by linear mixed model. Results Fatty acid desaturase (FADS) 1 activity decreased by 29.1% (P = 0.007) when the 6-mg Zn/d diet was consumed for 2 wk. This change was associated with significant decreases in HDL and LDL cholesterol. The alterations in FADS1, HDL cholesterol, and TG remained unchanged when Zn intakes were increased to 10 mg/d for 4 wk. Supplementation with 25 mg Zn/d for 3 wk normalized these metabolic changes and significantly increased LDL cholesterol at the end of this metabolic period compared with baseline. FADS1 activity was inversely correlated with FADS2 (r(mcorr) = -0.52; P = 0.001) and TG (r(mcorr) = -0.55; P = 0.001) at all time points. Conclusions A low-zinc, high-phytate rice-based diet reduced plasma HDL cholesterol concentrations and altered fatty acid profiles in healthy men within 2 wk. Consuming 10 mg Zn/d without phytate for 4 wk did not improve the lipid profiles, but a 25-mg Zn/d supplement corrects these alterations in lipid metabolism within 3 wk.

Excess dietary fat, and associated bile acids, can impair intestinal barrier integrity, produce intestinal or systemic inflammation and promote tumorigenesis. Dietary polyphenols in foods such as berries display antioxidant and other protective effects in many biological systems, but little is known about their protective effects on intestinal epithelial cells exposed to dietary fat. In a Caco-2 cell model of dietary fat-induced intestinal epithelial cell cytotoxicity, oxidative stress and barrier impairment, we investigated the relative protection afforded by an anthocyanin-rich bilberry extract (ARBE) or resveratrol. Exposure of the cells to mixed micelles (MM) of fatty acids and bile acids for 24 h markedly increased intracellular reactive oxygen species (ROS) and mitochondrial superoxide generation, decreased cell viability, increased expression of TNF-α mRNA and disrupted differentiated monolayer integrity. Starting prior to exposure to MM, treatments with ARBE or resveratrol, at polyphenol concentrations from 1.25-20 μM, strongly attenuated MM-induced intracellular ROS generation, and ARBE but not resveratrol decreased mitochondrial superoxide generation. Both ARBE and resveratrol inhibited the MM-induced expression of TNF-α mRNA. In assessments of differentiated monolayer integrity by transepithelial electrical resistance (TEER) and paracellular permeability, resveratrol protection was apparent at 3 h of MM exposure, but less at 6 h and absent by 9 h. In contrast, ARBE largely reversed MM-induced impairment by 9 h, with TEER values reaching 82% of control and the MM-induced paracellular permeability reduced by 78%. While they appeared to act differently, the results suggest that dietary sources of anthocyanins and resveratrol can help confer resistance of intestinal epithelial cells to oxidative stress and inflammation, and resultant barrier dysfunction and tumorigenesis, induced by high dietary fat common in the Western diet.

BACKGROUND:Poor diets contribute to metabolic complications of obesity, insulin resistance and dyslipidemia. Metabolomic biomarkers may serve as early nutrition-sensitive health indicators. This family-based lifestyle change program compared metabolic outcomes in an intervention group (INT) that consumed 2 nutrient bars daily for 2-months and a control group (CONT).METHODS:Overweight, predominantly minority and female adolescent (Teen)/parent adult caretaker (PAC) family units were recruited from a pediatric obesity clinic. CONT (8 Teen, 8 PAC) and INT (10 Teen, 10 PAC) groups randomized to nutrient bar supplementation attended weekly classes that included group nutrition counseling and supervised exercise. Pre-post physical and behavioral parameters, fasting traditional biomarkers, plasma sphingolipids and amino acid metabolites were measured.RESULTS:In the full cohort, a baseline sphingolipid ceramide principal component composite score correlated with adiponectin, triglycerides, triglyceride-rich very low density lipoproteins, and atherogenic small low density lipoprotein (LDL) sublasses. Inverse associations were seen between a sphingomyelin composite score and C-reactive protein, a dihydroceramide composite score and diastolic blood pressure, and the final principal component that included glutathionone with fasting insulin and the homeostatic model of insulin resistance. In CONT, plasma ceramides, sphinganine, sphingosine and amino acid metabolites increased, presumably due to increased physical activity. Nutrient bar supplementation (INT) blunted this rise and significantly decreased ureagenic, aromatic and gluconeogenic amino acid metabolites. Metabolomic changes were positively correlated with improvements in clinical biomarkers of dyslipidemia.CONCLUSION:Nutrient bar supplementation with increased physical activity in obese Teens and PAC elicits favorable metabolomic changes that correlate with improved dyslipidemia. The trial from which the analyses reported upon herein was part of a series of nutrient bar clinical trials registered at clinicaltrials.gov as NCT02239198.

Upon completion, participants will be able to understand the power of micronutrients in food-based, targeted, dietary interventions to treat obesity and associated metabolic dysregulation.

Asthma in the obese is often severe, difficult to treat, and characterized by less eosinophilic inflammation than asthma in the nonobese. Obesity-associated metabolic dysregulation may be a causal factor. We previously reported that a nutrient- and fiber-dense bar [Children's Hospital Oakland Research Institute (CHORI)-bar], which was designed to fill gaps in poor diets, improved metabolism in healthy overweight/obese (OW/OB) adults. In this pilot trial, OW/OB adolescents with poorly controlled asthma were randomized to weekly nutrition/exercise classes with or without twice-daily CHORI-bar consumption. Intent-to-treat analysis did not indicate CHORI-bar-specific effects. However, restricting the analysis to participants with acceptable compliance and a relatively low fraction of exhaled nitric oxide (FENO; <50/ ppb, a surrogate for noneosinophilic asthma; study participants: CHORI-bar, n = 16; controls, n = 15) indicated that CHORI-bar-specific, significant improvements in lung function (forced vital capacity, percent-predicted forced expiratory volume in 1 s, and percent-predicted forced expiratory flow between 25 and 75% of forced vital capacity), primarily in participants with low chronic inflammation (high-sensitivity C-reactive protein <1.5 mg/L). (We previously observed that chronic inflammation blunted CHORI-bar-induced metabolic improvements in healthy OW/OB adults.) Lung function improvement occurred without weight loss and was independent of improvements in metabolic and anthropometric end points and questionnaire-based measures of asthma control and quality of life. This study suggests that a nutritional intervention can improve lung function in OW/OB adolescents with asthma and relatively low FENO without requiring major changes in dietary habits, lifestyle, or weight loss and that this effect is blunted by chronic inflammation.-Bseikri, M., McCann, J. C., Lal, A., Fong, E., Graves, K., Goldrich, A., Block, D., Gildengoren, G. L., Mietus-Snyder, M., Shigenaga, M., Suh, J., Hardy, K., Ames, B. N. A novel nutritional intervention improves lung function in overweight/obese adolescents with poorly controlled asthma: the Supplemental Nutrition in Asthma Control (SNAC) pilot study.

Objective To measure biomarkers of lipid metabolism in response to a marginal zinc depletion, repletion, and supplementation in healthy male subjects. Methods Eighteen male subjects between ages of 18–45 consumed a controlled diet (80% carbohydrate, 10% fat, 10% protein) with two levels of dietary zinc. Phase I: Low zinc: 6 mg/day with 1.5 mg of phytate (2 weeks); Phase II: Zinc repletion: 10 mg/day with no phytate (4 weeks). Thereafter, a 25 mg/day zinc supplement was administered with an ad libitum diet for 3 weeks (Phase III). Targeted analysis of lipids and lipoprotein particle size were performed using standard methodology. Results Plasma zinc levels remained unchanged during Phases I and II in all subjects; they increased by 17.4 ± 3.8% (mean ± SE) at the end of supplementation (phase III). Apo‐A1 decreased significantly from 119.2 ± 13.2 mg/dL (mean ± SD) to 110.2 ± 13.2 mg/dL (p=0.003) at the end of the low zinc period. Apo‐A1 remained low at the end of phase II, but returned back to baseline after supplementation. A similar pattern was observed in HDL‐c and HDL‐L (large buoyant HDL particles) levels across the three phases. Plasma triglyceride levels increased during the 6‐week high‐carbohydrate diet, but were not modified by shifts in dietary zinc. Based on our previous observation of higher DNA strand breaks in the low zinc metabolic period and literature reports linking cancer to low HDL‐c levels, we performed a correlation analysis of DNA strand breaks and HDL subspecies. Our results show that the percent changes in HDL‐L and DNA strand breaks from phase I to II are negatively correlated (r=−0.45; p=0.05). Conclusion Results from our studies show that low zinc combined with a high carbohydrate diet increased biomarkers of dyslipidemia, but provision of adequate dietary zinc or a zinc supplement for a short period of time mitigated the increase in these biomarkers. Support or Funding Information Harvest Plus

BACKGROUND:Food fortification has been recommended to improve a population's micronutrient status. Biofortification techniques modestly elevate the zinc content of cereals, but few studies have reported a positive impact on functional indicators of zinc status.OBJECTIVE:We determined the impact of a modest increase in dietary zinc that was similar to that provided by biofortification programs on whole-body and cellular indicators of zinc status.DESIGN:Eighteen men participated in a 6-wk controlled consumption study of a low-zinc, rice-based diet. The diet contained 6 mg Zn/d for 2 wk and was followed by 10 mg Zn/d for 4 wk. To reduce zinc absorption, phytate was added to the diet during the initial period. Indicators of zinc homeostasis, including total absorbed zinc (TAZ), the exchangeable zinc pool (EZP), plasma and cellular zinc concentrations, zinc transporter gene expression, and other metabolic indicators (i.e., DNA damage, inflammation, and oxidative stress), were measured before and after each dietary-zinc period.RESULTS:TAZ increased with increased dietary zinc, but plasma zinc concentrations and EZP size were unchanged. Erythrocyte and leukocyte zinc concentrations and zinc transporter expressions were not altered. However, leukocyte DNA strand breaks decreased with increased dietary zinc, and the level of proteins involved in DNA repair and antioxidant and immune functions were restored after the dietary-zinc increase.CONCLUSIONS:A moderate 4-mg/d increase in dietary zinc, similar to that which would be expected from zinc-biofortified crops, improves zinc absorption but does not alter plasma zinc. The repair of DNA strand breaks improves, as do serum protein concentrations that are associated with the DNA repair process. This trial was registered at clinicaltrials.gov as NCT02861352.

ObjectiveTo determine the effects of marginal zinc deficiency and repletion on essential fatty acid, sphingolipid, and lipoprotein metabolism.MethodsSixteen apparently healthy male subjects between ages of 18–45 were subjected to three sequential phases of dietary zinc intake modulation: Phase 1: Low Zinc intake: 6 mg/day with 1.5 g of phytate (2 wks); Phase II: Zinc Repletion: 10 mg/day with no phytate (4 wks); Phase III: Zinc Supplementation: Ad lib diet supplemented with 20 mg zinc (2 wks). Subject weight, and compliance with diet were monitored every 3–4 days during the first two phases. Plasma free fatty acids and sphingolipids were measured by mass spectrometry. FADS1 and FADS2 activities were calculated by g‐linolenic (GLA)/linolenic acid (LA) or arachidonic acid (AA)/dihomo‐g‐linolenic acid (DGLA) ratios, respectively.ResultsPlasma zinc levels remained unchanged during Phases I and II in all subjects and increased only at the conclusion of phase III. FADS1 activity significantly (p=0.005) decreased from 5.2 ± 0.5 to 3.6 ± 0.5 (mean ± SEM) following Zn depletion. In contrast, FADS2 activity was insensitive to effects of low zinc intake. As a consequence of FADS1 activity loss, concentrations of AA‐containing phosphatidylcholine (PC), phosphatidylethanolamine (PE), sphingomyelins and plasmalogen antioxidant lipids decreased significantly. Furthermore, plasma concentrations of C16:0 ceramide increased following Zn depletion (p = 0.06) and was inversely correlated with FADS1 activity (p = 0.04; r2 = 0.6). Zn depletion also increased plasma triglyceride (TG) by 26 ± 15% (p< 0.05) and decreased high‐density lipoprotein (HDL) by 18.5 ± 3%; P<0.05. Interestingly, changes in FADS1 activity observed during the depletion was not normalized by dietary Zn repletion (Phase II) but, it was completely normalized with Zn supplementation (20 mg/day; Phase III).ConclusionDietary Zn depletion (6 mg/day) decreases FADS1 activity and lowered AA incorporation into major lipid sub‐classes. Loss in FADS1 activity was inversely correlated with C16:0 ceramide and was further associated with dyslipidemia as evidenced by significant alterations in plasma TG and HDL. These effects were not corrected with dietary Zn repletion (10 mg/day); supplementation with 20 mg Zn/d for an additional 3 weeks was required. These data suggest that FADS1 activity may be a sensitive biomarker of inadequate zinc intake and they also implicate marginal Zn intake as a novel risk factor for dyslipidemia and insulin resistance.Support or Funding InformationHarvest Plus, NIH S10OD0018070‐01

This study determined if twice-daily consumption of a nutrient-dense bar intended to fill gaps in Western diets, without other dietary/lifestyle requirements, favorably shifted metabolic/anthropometric indicators of dysregulation in a healthy direction. Three 8-wk clinical trials in 43 healthy lean and overweight/obese (OW/OB) adults, who served as their own controls, were pooled for analysis. In less inflamed OW/OB [high-sensitivity C-reactive protein (hsCRP) <1.5], statistically significant decreases occurred in weight (-1.1 ± 0.5 kg), waist circumference (-3.1 ± 1.4 cm), diastolic blood pressure (-4.1 ± 1.6 mmHg), heart rate [HR; -4.0 ± 1.7 beats per minute (bpm)], triglycerides (-72 ± 38.2 mg/dl), insulin resistance (homeostatic model of insulin resistance) (-0.72 ± 0.3), and insulin (-2.8 ± 1.3 mU/L); an increase in HDL-2b (+303 ± 116 nM) and realignment of LDL lipid subfractions toward a less atherogenic profile [decreased small LDL IIIb (-44 ± 23.5 nM), LDL IIIa (-99 ± 43.7 nM), and increased large LDL I (+66 ± 28.0 nM)]. In the more inflamed OW/OB (hsCRP >1.5), inflammation was reduced at 2 wk (-0.66 mg/L), and HR at 8 wk (-3.4 ± 1.3 bpm). The large HDL subfraction (10.5-14.5 nm) increased at 8 wk (+346 ± 126 nM). Metabolic improvements were also observed in lean participants. Thus, favorable changes in measures of cardiovascular health, insulin resistance, inflammation, and obesity were initiated within 8 wk in the OW/OB by replacing deficiencies in Western diets without requiring other dietary or lifestyle modifications; chronic inflammation blunted most improvements.

The ultimate function of an organism’s physiology is the performance of complex behaviors that facilitate reproduction. Here we use Organismal Performance Assays (OPAs) to quantify declines in Darwinian fitness of house mice due to experimental exposures. OPAs are sensitive phenotyping approaches that use semi‐natural conditions to challenge the physiology of differentially treated animals in direct competition with each other. Using OPAs we demonstrated that consumption of human‐relevant levels of added sugar (25% kcal) decreases reproduction of male mice by 25% while doubling the mortality rate of females relative to starch‐fed controls. Furthermore, we showed that many clinical measures associated with proximate effects of added sugar consumption were not predictive of organismal‐level effects. We expand upon these studies by using OPAs to test for differential health consequences of the two most common forms of added sugar. We report that females fed a diet containing an equal ratio of fructose and glucose monosaccharides (modeling high fructose corn syrup; HFCS) experienced a mortality rate 1.9 times higher and produced 26% fewer offspring than those fed sucrose (fructose‐glucose disaccharide). This experiment provides evidence that fructose and glucose monosaccharides (HFCS) are more deleterious to mammalian health than isocaloric sucrose.Grant Funding Source: Supported by NIH and NSF

Zinc deficiency continues to be a major concern of international nutrition, yet there are still few sensitive and reliable biomarkers for identifying moderate zinc deficiency or monitoring changes in zinc status. Plasma zinc content is used most frequently, but it is influenced by conditions other than zinc intake and, therefore, is not reliable except when zinc intake is very low. In this study, the sensitivity of leukocytic zinc transporter gene expression and genomic integrity were evaluated in a moderate depletion/repletion study. Eighteen healthy men were provided a low zinc (6 mg/day) diet with added phytate for two weeks followed by an adequate zinc (10 mg/day) diet for four weeks. They then consumed an ad libitum diet while taking a 25mg zinc supplement for three weeks. Leukocyte DNA damage increased (p<0.0001) during the depletion phase, as detected by the Comet Assay. ZIP1, ZIP4, and ZnT1 zinc transporter gene expression was evaluated in isolated leukocytes and measured using qPCR. ZnT1 gene expression increased (p=0.02) during the depletion phase, although total plasma and leukocyte zinc levels did not change. These results suggest that a short‐term, low zinc intake alters genomic integrity and zinc transporter expression, but not plasma zinc. These potential biomarkers may be useful to evaluate zinc deficiency and the effectiveness of zinc interventions in conditions where plasma zinc remains unchanged.Grant Funding Source: This study was funded by a grant from HarvestPlus.

Objective: To determine whether twice daily intake of a low-calorie (110 kcal), high-fiber, fruit and dark chocolate based nutrient-dense bar with supplemental vitamins/minerals, β-glucan, and docosahexaenoic acid might serve as an effective adjunct to lifestyle counseling for weight management in an inner city population. Methods: 18 overweight, predominantly female adolescent/parent guardian dyads and 2 triads (21 adults, 22 teens, randomized as 12 intervention (INT, with bars), 8 control (C) family units were recruited from a pediatric obesity clinic. The cohort was 48.7% Nonhispanic Black, 34.1% Hispanic, and 17.0% Caucasian. Two adults dropped out. The remaining 41 subjects participated in six identical weekly exercise and nutrition sessions by group. Full assessment of physical (BMI, blood pressure), behavioral (diet, activity), metabolic (cardiovascular and diabetes risk biomarkers), and metabolomic status was conducted at baseline and end-of-study. Results: There was excellent attendance in both INT and C groups with all family units participating in more than 80% of group sessions and 100% of baseline and follow-up assessment visits. Compliance with nutrition bar intake was 86 ± 11% and 87 ± 14% among INT group adults and teens respectively. There was considerable obesity, hypertension, dyslipidemia, inflammation, and insulin resistance in all subjects and baseline diets were universally poor. Self-report activity increased and dietary habits improved in both INT and C groups, adults and teens, most notably decreased saturated fat and total carbohydrates (especially added sugars). Weight was stable, even in the INT group despite the addition of 220 additional kcal in two daily nutrition bars. In paired analyses, systolic blood pressure (SBP) improved significantly in INT teens (-6.7 ± 9.2 mm Hg, p = 0.02), and worsened in C teens (+6.3 ± 7.7 mm Hg, p = 0.04); p = 0.002 for unpaired comparison by teen group. Plasma homocysteine levels and two amino acids, citrulline and sarcosine, implicated in the arginine-urea cycle pathway fell in both INT parents and teens, but not in controls. The drop in citrulline was more significant in INT teens ( p = 0.005) than in adults ( p = 0.047). In a diet-induced obesity mouse model, elevated plasma citrulline has been associated with cardiometabolic complications attributed to decreased systemic arginine bioavailability that may affect capacity to produce nitric oxide. The observed metabolomic changes are consistent with more efficient mitochondrial processing and may relate to the systolic BP improvement observed in teens. Conclusions: A nutritional supplement bar may be a valuable adjunct for weight management, resulting in early favorable metabolomic changes.

Consumption of added sugar has increased over recent decades and is correlated with numerous diseases. Rodent models have elucidated mechanisms of toxicity, but only at concentrations beyond typical human exposure. Here we show that comparatively low levels of added sugar consumption have substantial negative effects on mouse survival, competitive ability, and reproduction. Using Organismal Performance Assays--in which mice fed human-relevant concentrations of added sugar (25% kcal from a mixture of fructose and glucose, modeling high fructose corn syrup) and control mice compete in seminatural enclosures for territories, resources and mates--we demonstrate that fructose/glucose-fed females experience a twofold increase in mortality while fructose/glucose-fed males control 26% fewer territories and produce 25% less offspring. These findings represent the lowest level of sugar consumption shown to adversely affect mammalian health. Clinical defects of fructose/glucose-fed mice were decreased glucose clearance and increased fasting cholesterol. Our data highlight that physiological adversity can exist when clinical disruptions are minor, and suggest that Organismal Performance Assays represent a promising technique for unmasking negative effects of toxicants.

Dietary intake modulates disease risk, but little is known how components within food mixtures affect pathophysiology. A low-calorie, high-fiber, fruit-based nutrient-dense bar of defined composition (e. g., vitamins and minerals, fruit polyphenolics, beta-glucan, docosahexaenoic acid) appropriate for deconstruction and mechanistic studies is described and evaluated in a pilot trial. The bar was developed in collaboration with the U. S. Department of Agriculture. Changes in cardiovascular disease and diabetes risk biomarkers were measured after 2 wk twice-daily consumption of the bar, and compared against baseline controls in 25 healthy adults. Plasma HDL-cholesterol (HDL-c) increased 6.2% (P = 0.001), due primarily to a 28% increase in large HDL (HDL-L; P<0.0001). Total plasma homocysteine (Hcy) decreased 19% (P = 0.017), and glutathione (GSH) increased 20% (P = 0.011). The changes in HDL and Hcy are in the direction associated with decreased risk of cardiovascular disease and cognitive decline; increased GSH reflects improved antioxidant defense. Changes in biomarkers linked to insulin resistance and inflammation were not observed. A defined food-based supplement can, within 2 wk, positively impact metabolic biomarkers linked to disease risk. These results lay the groundwork for mechanistic/deconstruction experiments to identify critical bar components and putative synergistic combinations responsible for observed effects.-Mietus-Snyder, M. L., Shigenaga, M. K., Suh, J. H., Shenvi, S. V., Lal, A., McHugh, T., Olson, D., Lilienstein, J., Krauss, R. M., Gildengoren, G., McCann, J. C., Ames, B. N. A nutrient-dense, high-fiber, fruit-based supplement bar increases HDL cholesterol, particularly large HDL, lowers homo-cysteine, and raises glutathione in a 2-wk trial. FASEB J. 26, 3515-3527 (2012). www.fasebj.org

Animal and human studies show that intracellular zinc levels are sensitive to changes in dietary zinc. The protein metallothionein (MT) is maintained in proportion to intracellular zinc content, and therefore may be a useful marker of zinc status. Measuring MT levels is difficult due to the unique properties of the protein, so most studies focus on MT gene expression. Our objective was to develop new methods to quantify MT protein and MT‐bound zinc content. Zinc deficiency or excess was modeled in the Jurkat leukocytic cell line using the zinc‐selective chelator TPEN or exogenous zinc sulfate, respectively. Cellular protein was separated using size‐exclusion filtration to isolate an MT‐rich fraction. Then, endogenous zinc content was released, apo‐MT was loaded with cadmium, and MT‐bound cadmium levels were quantified by ICP; this method permitted the determination of cellular MT‐bound zinc content (MBZC) and MT zinc saturation (%MTsat). Gene expression of the major MT and zinc transporter isoforms were measured in parallel by RT‐PCR, which reflected the change in zinc balance in these cells. MBZC and %MTsat levels correlated with intracellular zinc content and changes in MT gene expression, showing bimodal responses to zinc deficiency and zinc excess. These measures of the cellular zinc‐metallothionein ratio may have utility as markers of zinc homeostasis. This work was supported by the HarvestPlus.

Achieving recommended dietary intakes of vitamins and minerals, fiber, and other health-promoting small molecules, such as fruit polyphenolics and essential fatty acids, is difficult, even for healthy adults consuming generally good diets. We assessed the hypothesis that short term consumption of a comprehensive nutrition bar can favorably impact metabolic biomarkers linked to future disease risk. Methods: An approximately 100 kcal nutrition bar containing a vitamin, mineral, and docosahexaenoic acid (DHA)-fortified polyphenolic-rich whole food matrix composed of fruit, walnuts, and dark chocolate blended with insoluble and soluble fiber was developed. Pooled results are presented from 3 pilot trials in which 25 generally healthy adults (15 F, 10 M) were enrolled in 2-wk interventions to study the effect of twice-daily consumption of the bar on plasma lipoprotein subfractions (ion mobility analysis), and redox status (thiol amino acids) in addition to conventional biomarkers associated with dyslipidemia, insulin resistance, and inflammation. All variables were continuous, and paired t-tests were used to test for significant change between baseline and follow-up measures. Results: Mean HDL cholesterol increased by 6.6 + 9.7% ( p = 0.001), predominantly due to a rise in the large buoyant HDL2b subclass which rose from 1665.9 ± 1017.7 to 2089.4 ± 1197.6 nmol/L ( p < 0.0001; 28% increase). Plasma triglyceride and LDL cholesterol levels did not change significantly but there was a trend towards increased LDL peak particle diameter (p = 0.086). Mean plasma homocysteine decreased from 12.6 ± 6.4 to 10.2 ± 3.7 µM (p=0.006, 19.2% decline). Measures of insulin resistance and inflammation did not change. Weight remained stable despite intake of 200 additional daily calories. Participants reported a significant decrease in hunger on an analog scale and compensatory decreased intake of other foods. Summary: Twice daily consumption of a low-calorie complete nutritional supplement increases satiety and large HDL particle concentrations within 2 weeks without weight gain.

Adverse effects of fructose at human‐relevant levels have been suspected from epidemiology, but empirical confirmation has been lacking. Here we use a novel methodology (OPAs) to demonstrate health declines from diets of free fructose and glucose (modeling HFCS) compared to diets of sucrose or no added sugar. During OPAs mice raised on experimental and control diets compete directly for resources, territories and mates in seminatural enclosures. Due to the competitive nature of mouse ecology, differences in individual performance result in quantitative fitness differentials. OPA data are reported from two studies comparing a free fructose and glucose diet (25 % Kcal) vs. either sucrose (25% Kcal) or a diet free of added sugar. OPAs revealed that females on the monosaccharide diet experience a 3‐fold increase in mortality compared to females raised on either sucrose or sugar‐free diets. Likewise, OPAs revealed that males on the sugar‐free diet obtain social dominance at 4‐times the rate and produce 50% more offspring than males fed simple sugar. These findings represent the lowest observed adverse effect level for dietary fructose (12.5% Kcal) and indicate that the free form of fructose is more deleterious than sucrose.