Methylenetetrahydrofolate reductase deficiency and high-dose FA supplementation disrupt embryonic development of energy balance and metabolic homeostasis in zebrafish.

Folic acid (synthetic folate, FA) is consumed in excess in North America and may interact with common pathogenic variants in methylenetetrahydrofolate reductase (MTHFR); the most prevalent inborn error of folate metabolism with wide-ranging obesity-related comorbidities. While preclinical murine models have been valuable to inform on diet-gene interactions, a recent Folate Expert panel has encouraged validation of new animal models. In this study, we characterized a novel zebrafish model of mthfr deficiency and evaluated the effects of genetic loss of mthfr function and FA supplementation during embryonic development on energy homeostasis and metabolism. mthfr-deficient zebrafish were generated using CRISPR mutagenesis and supplemented with no FA (control, 0FA) or 100 µm FA (100FA) throughout embryonic development (0-5 days postfertilization). We show that the genetic loss of mthfr function in zebrafish recapitulates key biochemical hallmarks reported in MTHFR deficiency in humans and leads to greater lipid accumulation and aberrant cholesterol metabolism as reported in the Mthfr murine model. In mthfr-deficient zebrafish, energy homeostasis was also impaired as indicated by altered food intake, reduced metabolic rate and lower expression of central energy-regulatory genes. Microglia abundance, involved in healthy neuronal development, was also reduced. FA supplementation to control zebrafish mimicked many of the adverse effects of mthfr deficiency, some of which were also exacerbated in mthfr-deficient zebrafish. Together, these findings support the translatability of the mthfr-deficient zebrafish as a preclinical model in folate research.

Potatoes Compared with Rice in Meals with either Animal or Plant Protein Reduce Postprandial Glycemia and Increase Satiety in Healthy Adults: A Randomized Crossover Study.

BACKGROUND: Rice and pasta are recommended as healthier than potatoes on the basis of their glycemic index when eaten alone. OBJECTIVES: The study objective was to evaluate postprandial glycemia (PPG), appetite, and food intake (FI) at meals with potatoes or rice when consumed with either meatballs or their vegetarian substitute. METHODS: In a randomized, single-blinded, crossover design, 26 (13 males and 13 females) healthy adults (age: 18-45 y; body mass index [kg/m2]: 18.5-29.9) consumed isocaloric fixed amounts of either meatballs or vegetarian-substitute balls with ad libitum access to either baked French fries (BFF), instant mashed potatoes (IMPs), or rice (control). FI was measured at the meal and at an ad libitum pizza meal served 120 min later. Blood glucose (BG), appetite, and plasma insulin responses were measured within the meal (0-30 min), postmeal (30-120 min), within pizza meal (120-140 min), and post-pizza (140-170 min). Effects of protein source, carbohydrate (CHO) source, and sex and their interactions were analyzed using analysis of variance followed by Tukey's post hoc test. RESULTS: Participants consumed 23-25% less treatment meal energy (kcal), 32-34% less CHO energy (kcal), and 13-16% less total energy (kcal) after the BFF and IMP than rice meals (P < 0.0001). Postmeal BG was lower after IMP (6.76 ± 0.15; P < 0.0001) and rice (6.92 ± 0.15; P = 0.0012) compared with BFF (7.19 ± 0.15). Post-pizza BG was higher after rice (6.77 ± 0.09) than that after BFF (6.51 ± 0.09; P = 0.0012) and IMP (6.39 ± 0.09; P < 0.0001). Postmeal meaned insulin was higher after BFF (82.16 ± 8.58) and IMP (77.75 ± 8.60) compared with rice (56.44 ± 8.59; P < 0.002). Insulin during pizza meal was lower after BFF (17.14 ± 6.90) compared with both IMP (39.03 ± 6.90; P = 0.0060) and rice (34.21 ± 6.90; P = 0.0336). Meatballs led to lower BG (6.48 ± 0.09; P = 0.0076) and higher insulin (84.54 ± 5.87; P = 0.0406) post-pizza compared with their plant protein substitute (6.64 ± 0.09 and 73.18 ± 5.87, respectively). CONCLUSIONS: Adults consuming meatballs or plant-based substitute with ad libitum IMP had lower PPG post-treatment and at a later pizza meal compared with rice. Both IMP and BFF resulted in lower energy intake than after rice. This trial was registered at http://clinicaltrials.gov (https://register. CLINICALTRIALS: gov/prs/app/action/SelectProtocol?sid=S000CKIJ&selectaction=Edit&uid=U0000IA4&ts=2&cx=-uf51kf) as NCT05610124. Protocol ID: 43406 (Postprandial Glycemia and Satiety of Meals with Potatoes, with and without Protein).

Age and Sex Interact to Determine the Effects of Commonly Consumed Dairy Products on Postmeal Glycemia, Satiety, and Later Meal Food Intake in Adults.

BACKGROUND: Dairy consumption reduces postprandial glycemia and appetite when consumed with carbohydrates. OBJECTIVES: The objective was to test the effects of frequently consumed dairy products, age, and sex on glycemia, appetite, and food intake. METHODS: In a randomized, unblinded, crossover design, 30 older [60-70 y; BMI (kg/m2): 18.5-29.9] and 28 young (20-30 y; BMI: 18.5-24.9) adults consumed 500 mL of a calorie-free control (water), skim milk and whole milk, 350 g Greek yogurt, and 60 g cheddar cheese. Food intake at an ad libitum meal was measured 120 min later. Glycemia, appetite, and gastric hormone responses were measured premeal (15-120 min), within-meal (120-140 min), and postmeal (140-170 min). Effects of treatment, age, and sex and their interactions were analyzed using ANOVA followed by Tukey's post hoc test. RESULTS: All forms of dairy, compared with water, decreased postmeal glycemia, premeal appetite, and meal intake (P < 0.0001). Premeal glucose, insulin, and glucagon-like peptide 1 increased, and ghrelin decreased, but effects of dairy differed with age and sex. Older adults had 10% higher pre- and postmeal glucose (P < 0.01). Premeal appetite suppression per 100 kcal of treatments was more after yogurt than other dairy, but overall appetite suppression was less in older adults than in young adults and in males than in females (P < 0.05). Pizza intake was reduced by 175 kcal after yogurt and cheese and by 82 kcal after milks compared to water (P < 0.001). Mealtime reduction for treatment calories averaged 62% after yogurt and cheese but was less at 33% after milks (P < 0.05). Compensation was less in older (33%) than in young (63%) adults (P < 0.03). CONCLUSIONS: Dairy products consumed in usual forms before a meal stimulate metabolic responses leading to reduced premeal appetite, later food intake, and postmeal glycemia, but their effects differ in magnitude and with the sex and age of adults.

Choline and Folic Acid in Diets Consumed during Pregnancy Interact to Program Food Intake and Metabolic Regulation of Male Wistar Rat Offspring.

BACKGROUND: North American women consume high folic acid (FA), but most are not meeting the adequate intakes for choline. High-FA gestational diets induce an obesogenic phenotype in rat offspring. It is unclear if imbalances between FA and other methyl-nutrients (i.e., choline) account for these effects. OBJECTIVE: This study investigated the interaction of choline and FA in gestational diets on food intake, body weight, one-carbon metabolism, and hypothalamic gene expression in male Wistar rat offspring. METHODS: Pregnant Wistar rats were fed an AIN-93G diet with recommended choline and FA [RCRF; 1-fold, control] or high (5-fold) FA with choline at 0.5-fold [low choline and high folic acid (LCHF)], 1-fold [recommended choline and high folic acid (RCHF)], or 2.5-fold [high choline and high folic acid (HCHF)]. Male offspring were weaned to an RCRF diet for 20 wk. Food intake, weight gain, plasma energy-regulatory hormones, brain and plasma one-carbon metabolites, and RNA sequencing (RNA-seq) in pup hypothalamuses were assessed. RESULTS: Adult offspring from LCHF and RCHF, but not HCHF, gestational diets had 10% higher food intake and weight gain than controls (P < 0.01). HCHF newborn pups had lower plasma insulin and leptin compared with LCHF and RCHF pups (P < 0.05), respectively. Pup brain choline (P < 0.05) and betaine (P < 0.01) were 22-33% higher in HCHF pups compared with LCHF pups; methionine was ∼23% lower after all high FA diets compared with RCRF (P < 0.01). LCHF adult offspring had lower brain choline (P < 0.05) than all groups and lower plasma 5-methyltetrahydrofolate (P < 0.05) than RCRF and RCHF groups. HCHF adult offspring had lower plasma cystathionine (P < 0.05) than LCHF adult offspring and lower homocysteine (P < 0.01) than RCHF and RCRF adult offspring. RNA-seq identified 144 differentially expressed genes in the hypothalamus of HCHF newborns compared with controls. CONCLUSIONS: Increased choline in gestational diets modified the programming effects of high FA on long-term food intake regulation, plasma energy-regulatory hormones, one-carbon metabolism, and hypothalamic gene expression in male Wistar rat offspring, emphasizing a need for more attention to the choline and FA balance in maternal diets.

Role of Amino Acids in Blood Glucose Changes in Young Adults Consuming Cereal with Milks Varying in Casein and Whey Concentrations and Their Ratio.

BACKGROUND: Increasing the total protein content and reducing the casein to whey ratio in milks consumed with breakfast cereal reduce postprandial blood glucose (BG). OBJECTIVES: We aimed to explore associations between plasma amino acids (AAs), BG, and glucoregulatory hormones. METHODS: In this repeated-measures design, 12 healthy adults consumed cereal (58 g) and milks (250 mL) with 3.1 wt% or high 9.3 wt% protein concentrations and with casein to whey ratios of either 80:20 or 40:60. Blood was collected at 0, 30, 60, 120, 140, 170, and 200 min for measurement of the primary outcome, BG, and for the exploratory outcomes such as plasma AA, gastric emptying, insulin (INS), and glucoregulatory hormones. Measures were made prior to and after an ad libitum lunch at 120 min. Exploratory correlations were conducted to determine associations between outcomes. RESULTS: Pre-lunch plasma AA groups [total (TAA), essential (EAA), BCAA, and nonessential (NEAA)] were higher after 9.3 wt% than 3.1 wt% milks by 12.7%, 21.4%, 20.9%, and 7.6%, respectively (P ≤ 0.05), while post-lunch AA groups were higher by 10.9%, 19.8%, 18.8%, and 6.0%, respectively (P ≤ 0.05). Except for NEAA, pre-lunch AAs were higher after 40:60 than 80:20 ratio milks by 4.5%, 8.3%, and 9.3% (P ≤ 0.05). When pooled by all treatments, pre-lunch AA groups associated negatively with BG (r/ρ ≥ -0.45, P ≤ 0.05), but post-lunch only TAA and NEAA correlated (r ≥ -0.37, P < 0.05). Pre-lunch BG was inversely associated with Leu, Ile, Lys, Met, Thr, Cys-Cys, Asn, and Gln (r/ρ ≥ -0.46, P ≤ 0.05), but post-lunch, only with Thr, Ala, and Gly (r ≥ -0.50, P ≤ 0.05). Pre-lunch associations between AA groups and INS were not found. CONCLUSIONS: Protein concentration and the ratio of casein to whey in milks consumed at breakfast with cereal affect plasma AA concentrations and their associations with decreased BG. The decrease in BG could be explained by INS-independent mechanisms. This trial was registered at www.clinicaltrials.gov as NCT02471092.

Gestational folic acid content alters the development and function of hypothalamic food intake regulating neurons in Wistar rat offspring post-weaning.

Background: Folic acid plays an important role in early brain development of offspring, including proliferation and differentiation of neural stem cells known to impact the function of food intake regulatory pathways. Excess (10-fold) intakes of folic acid in the gestational diet have been linked to increased food intake and obesity in male rat offspring post-weaning.Objective: The present study examined the effects of folic acid content in gestational diets on the development and function of two hypothalamic neuronal populations, neuropeptide Y (NPY) and pro-opiomelanocortin (POMC), within food intake regulatory pathways of male Wistar rat offspring at birth and post-weaning.Results: Folic acid fed at 5.0-fold above recommended levels (5RF) to Wistar dams during pregnancy increased the number of mature NPY-positive neurons in the hypothalamus of male offspring, compared to control (RF), 0RF, 2.5RF, and 10RF at birth. Folic acid content had no effect on expression and maturation of POMC-positive neurons. Body weight and food intake were higher in all treatment groups (2.5-, 5.0-, and 10.0-fold folic acid) from birth to 9 weeks post-weaning compared to control. Increased body weight and food intake at 9-weeks post-weaning were accompanied by a reduced activation of POMC neurons in the arcuate nucleus (ARC).Conclusion: Gestational folic acid content modulates expression of mature hypothalamic NPY-positive neurons at birth and activation of POMC-positive neurons at 9-weeks post-weaning in the ARC of male Wistar rat offspring which may contribute to higher body weight and food intake later in life.

Increased milk protein content and whey-to-casein ratio in milk served with breakfast cereal reduce postprandial glycemia in healthy adults: An examination of mechanisms of action.

This study describes the effects on glycemic response and the underlying mechanisms of action of increasing the protein concentration and decreasing the casein-to-whey ratio in milk when consumed with a high glycemic breakfast cereal. Twelve healthy men and women, aged 18 to 30 yr and with a body mass index of 20 to 24.9 kg/m2, consumed (in random order) milk beverages (250 mL) containing either 3.1 or 9.3% protein and casein-to-whey ratios of either 80:20 or 40:60. We measured postprandial appetite, glucose, regulatory hormones, and stomach emptying rate over 200 min, as well as food intake at an ad libitum meal at 120 min. Although pre-meal appetite was suppressed to a greater extent with milk beverages that had high (9.3%) compared with regular (3.1%) protein content, food intake was similar among all 4 treatments. Pre-meal mean blood glucose was lower with beverages that had high rather than regular milk protein content, with the lowest glucose peaks after the high milk protein treatment with the 40:60 casein-to-whey ratio. Pre-meal insulin and C-peptide levels were not affected by milk protein content or casein-to-whey ratio, but pre-meal glucagon-like peptide 1 was higher after the treatment containing high milk protein and the 40:60 casein-to-whey ratio, and pre-meal cholecystokinin was higher after the treatments containing high milk protein content. Plasma paracetamol response was also lower after the treatments containing high compared with regular milk protein content. When consumed with carbohydrate, milk beverages with high protein content and (to a lesser extent) a decreased casein-to-whey ratio lowered postprandial glycemia through insulin-independent mechanisms, primarily associated with delayed stomach emptying.

Acute effects of monosodium glutamate addition to whey protein on appetite, food intake, blood glucose, insulin and gut hormones in healthy young men.

AIMS: This study investigated the effects of adding monosodium glutamate (MSG) to carrot soup with or without whey protein, on subjective appetite, food intake (FI) and satiety hormones in healthy young men. METHODS: Two experiments were conducted using a repeated-measures, within-subject, crossover design. In exp-1 healthy young men (n = 28) consumed water alone (500 mL), or carrot soup (500 g) with or without MSG (5 g, 1% w/w) or whey protein enriched (36 g) carrot soup with or without MSG (5 g, 1% w/w). Subjective appetite was measured post-treatment and FI measured at a meal at 120 min. In exp-2 (n = 15) the same treatments except for water were used. In addition to subjective appetite and FI, blood glucose, insulin, glucose like peptide 1 (GLP-1), C-peptide and ghrelin were measured. RESULTS: Adding MSG to carrot soup or whey protein enriched carrot soup did not affect FI. However, in exp-1 the addition of both MSG and protein increased fullness, and when MSG was added to carrot soup reduced desire to eat. In exp-2, average post-treatment appetite (5-120 min) was lower after carrot soup with MSG and protein than all other treatments (P < 0.05). In exp-2, carrot soup with MSG and protein, but not with protein alone, increased post-treatment insulin and C-peptide, and lowered blood glucose in comparison to carrot soup with no additions (P < 0.05). CONCLUSION: Adding MSG alone, or in combination with whey protein, to carrot soups did not affect FI. However, MSG increased fullness and reduced desire to eat, as well as subjective appetite, and when added to protein decreased blood glucose and increased insulin and C-peptide, offering some support for the hypothesis that MSG in the gut signals protein consumption.

Acute decrease in serum testosterone after a mixed glucose and protein beverage in obese peripubertal boys.

BACKGROUND AND OBJECTIVES: Delayed puberty and lower levels of testosterone (T) have been observed in adult obese males and some adolescent males. In adult men, enteral glucose ingestion results in acute lowering of serum testosterone levels; however, this has not been studied in adolescents. We aimed to examine the acute effect of a glucose/protein beverage on serum T concentration changes in obese peripubertal males. A second objective was to determine whether change in T concentration was related to appetite hormone levels. PATIENTS AND METHODS: Twenty-three overweight and obese males aged 8-17 in pre-early (Tanner stage 1-2) and mid-late (Tanner stage 3-5) puberty were included in this cross-sectional study at the Clinical investigative unit at the Hospital for Sick Children. Participants consumed a beverage containing glucose and protein, and blood samples measuring pubertal hormones, ghrelin and glucagon-like peptide-1 (GLP-1) were taken over 60 min. RESULTS: Across pubertal stages, there was a significant decrease in T levels in adolescent boys (-18·6 ± 3·1%, P < 0·01) with no proportional differences between pre-early and mid-late puberty (P = 0·09). Decrease in T was associated with a decrease in LH (r = 0·52, P = 0·02), and fasting T was inversely correlated with fasting ghrelin (r = -0·51, P = 0·03) with no correlation with GLP-1. CONCLUSIONS: Intake of a mixed glucose/protein beverage acutely decreases T levels in overweight and obese peripubertal boys. A potential mechanism for this decrease may be secondary to an acute decrease in LH, but this requires further evaluation.

Canned Navy Bean Consumption Reduces Metabolic Risk Factors Associated with Obesity.

The high prevalence of obesity and its metabolic co-morbidities require dietitians to promote lifestyle modifications that can be effectively implemented into practice and are feasible for customers to adhere to. The objective of this study was to determine the effect of commercially available ready-to-eat canned navy beans added to the habitual diet on risk factors associated with obesity. Fourteen overweight and obese adults consumed 5 cups of canned navy beans per week for 4 weeks. The study results demonstrated that bean consumption results in reduced waist circumference in females by 2.5 cm and males by 2.1 cm (P < 0.001). The effect of beans on pulse rate, total cholesterol (TC), and low-density lipoprotein cholesterol (LDL) were sex dependent (P < 0.05). In males, pulse rate, TC, and LDL were decreased by 6.5%, 11.5%, and 18%, respectively. In females, pulse rate increased by 9.6%, and TC and LDL were relatively unchanged. There was a trend for a decreased glucose AUC (P = 0.06) in response to a glucose load. This study demonstrates that consuming 5 cups per week of ready-to-eat canned navy beans for 4 weeks reduces metabolic risk factors associated with obesity and therefore can be used as a tool in dietetic practice.

Obesity, sex and pubertal status affect appetite hormone responses to a mixed glucose and whey protein drink in adolescents.

BACKGROUND AND OBJECTIVES: Little information is available on how food intake regulatory hormones may be altered during pubertal development and across the weight spectrum in adolescents. Therefore, the effect of obesity, sex and pubertal status on subjective appetite and appetite hormones in response to a mixed glucose and whey protein drink was determined in 8-18 year old adolescents. PATIENTS AND METHODS: A cross-sectional cohort study was conducted at the Hospital for Sick Children, Toronto. After a 12 h fast, normal weight (n = 5 female, 4 male) and obese (n = 5 female, 4 male) adolescents (Experiment 1), and pre-early pubertal (n = 10) and mid-late pubertal (n = 10) obese male adolescents (Experiment 2) consumed a 250 ml glucose (30 g) and whey protein (30 g) beverage. Insulin, PYY, ghrelin and subjective appetite were measured over 120 min. RESULTS: Obese adolescents (Experiment 1) have higher insulin, PYY and lower ghrelin (P < 0·006) than normal weight controls, with a more pronounced effect in males (P < 0·037). Puberty (Experiment 2) did not affect insulin (P = 0·305), but the change in PYY in response to the drink was greater (P = 0·032) and ghrelin was lower (P = 0·026) in mid-late pubertal than pre-early pubertal obese males. Average appetite 60 min post-drink was higher in obese and mid-late pubertal adolescents, but not related to hormone changes. CONCLUSIONS: Obesity, sex and pubertal status affect macronutrient-stimulated appetite hormone secretion and these factors may alter food intake in obese children during pubertal development.

The acute effect of commercially available pulse powders on postprandial glycaemic response in healthy young men.

Whole pulses (beans, peas, chickpeas and lentils) elicit low postprandial blood glucose (BG) responses in adults; however, their consumption in North America is low. One potential strategy to increase the dietary intake of pulses is the utilisation of commercial pulse powders in food products; however, it is unclear whether they retain the biological benefits observed with whole pulses. Therefore, the present study examined the effects of commercially prepared pulse powders on BG response before and after a subsequent meal in healthy young men. Overall, three randomised, within-subject experiments were conducted. In each experiment, participants received whole, puréed and powdered pulses (navy beans in Expt 1; lentils in Expt 2; chickpeas in Expt 3) and whole-wheat flour as the control. All treatments were controlled for available carbohydrate content. A fixed-energy pizza meal (50·2 kJ/kg body weight) was provided at 120 min. BG concentration was measured before (0-120 min) and after (140-200 min) the pizza meal. BG concentration peaked at 30 min in all experiments, and pulse forms did not predict their effect on BG response. Compared with the whole-wheat flour control, navy bean treatments lowered peak BG concentrations (Expt 1, P< 0.05), but not the mean BG concentration over 120 min. The mean BG concentration was lower for all lentil (Expt 2, P= 0.008) and chickpea (Expt 3, P= 0.002) treatments over 120 min. Processing pulses to powdered form does not eliminate the benefits of whole pulses on BG response, lending support to the use of pulse powders as value-added food ingredients to moderate postprandial glycaemic response.

Recent advances in dietary proteins and lipid metabolism.

PURPOSE OF REVIEW: There is growing interest in defining the dietary approaches for the management of lipid disorders. This review focuses on dietary proteins. RECENT FINDINGS: Increasing protein intake was coupled with improved lipid profiles in humans and animals. However, most studies increased the protein content by reducing that of fat or carbohydrate or both, making unclear the role of protein alone. Mechanisms of action differ with the sources of proteins, because of amino acid composition and bioactive peptides encrypted in their sequences. Soy protein was investigated the most, and many studies show that its consumption reduces blood cholesterol. The role of other constituents including isoflavones is debated. Short-term studies are consistent in showing lipid-lowering properties of whey proteins, attributed to their relative high content in branched chain amino acids. A limited number of studies, the majority being on animals, have shown hypocholesterolemic activities of fish proteins. SUMMARY: Dietary proteins regulate lipid metabolism in a manner dependent on their quantity and composition. There is a general consensus that proteins slow lipid absorption and synthesis, and promote lipid excretion. The benefits of dietary proteins remain to be confirmed in individuals with lipid abnormalities, for formulation of optimal dietary alternatives for the management of lipid disturbances.

The Effect of Micronutrients on Obese Phenotype of Adult Mice Is Dependent on the Experimental Environment.

The environment of the test laboratory affects the reproducibility of treatment effects on physiological phenotypes of rodents and may be attributed to the plasticity of the epigenome due to nutrient-gene-environment interactions. Here, we explored the reproducibility of adding a multi-vitamin-mineral (MVM) mix to a nutrient-balanced high-fat (HF) diet on obesity, insulin resistance (IR), and gene expression in the tissues of adult male mice. Experiments of the same design were conducted in three independent animal facilities. Adult C57BL/6J male mice were fed an HF diet for 6 weeks (diet induced-obesity model) and then continued for 9-12 weeks on the HF diet with or without 5-fold additions of vitamins A, B1, B6, B12, Zn, and 2-fold Se. The addition of the MVM affected body weight, fat mass, gene expression, and markers of IR in all three locations (p < 0.05). However, the direction of the main effects was influenced by the interaction with the experimental location and its associated environmental conditions known to affect the epigenome. In conclusion, MVM supplementation influenced phenotypes and expression of genes related to adipose function in obese adult male mice, but the experimental location and its associated conditions were significant interacting factors. Preclinical studies investigating the relationship between diet and metabolic outcomes should acknowledge the plasticity of the epigenome and implement measures to reproduce studies in different locations.

Serum measures of docosahexaenoic acid (DHA) synthesis underestimates whole body DHA synthesis in male and female mice.

Females have higher docosahexaenoic acid (DHA) levels than males, proposed to be a result of higher DHA synthesis rates from α-linolenic acid (ALA). However, DHA synthesis rates are reported to be low, and have not been directly compared between sexes. Here, we apply a new compound specific isotope analysis model to determine n-3 PUFA synthesis rates in male and female mice and assess its potential translation to human populations. Male and female C57BL/6N mice were allocated to one of three 12-week dietary interventions with added ALA, eicosapentaenoic acid (EPA) or DHA. The diets included low carbon-13 (δ13C)-n-3 PUFA for four weeks, followed by high δ13C-n-3 PUFA for eight weeks (n=4 per diet, time point, sex). Following the diet switch, blood and tissues were collected at multiple time points, and fatty acid levels and δ13C were determined and fit to one-phase exponential decay modeling. Hepatic DHA synthesis rates were not different (P>.05) between sexes. However, n-3 docosapentaenoic acid (DPAn-3) synthesis from dietary EPA was 66% higher (P<.05) in males compared to females, suggesting higher synthesis downstream of DPAn-3 in females. Estimates of percent conversion of dietary ALA to serum DHA was 0.2%, in line with previous rodent and human estimates, but severely underestimates percent dietary ALA conversion to whole body DHA of 9.5%. Taken together, our data indicates that reports of low human DHA synthesis rates may be inaccurate, with synthesis being much higher than previously believed. Future animal studies and translation of this model to humans are needed for greater understanding of n-3 PUFA synthesis and metabolism, and whether the higher-than-expected ALA-derived DHA can offset dietary DHA recommendations set by health agencies.

A Comparison of the Effects of Dairy Products with their Plant-based Alternatives on Metabolic Responses in Healthy Young Canadian Adults: a randomized cross-over study.

Plant-based food demand is rapidly increasing. However, the metabolic responses of plant proteins within their commercially available form remains unclear. Two randomized crossover experiments compared plant-based alternatives to dairy on postprandial glycemia, metabolic hormones, and appetite before and after a fixed size (12 kcal/kg body weight) pasta meal in sixteen healthy young adults (eight males and eight females). In experiment one, participants (22.8±2.3y) consumed one-serving of Greek yogurt (175g), cheddar cheese (30g), plant-based cheese (30g), or plant-based yogurt (175g). In experiment two, participants (22.3±2.4y) consumed one-serving (250 mL) of cow's milk, vanilla soy beverage or vanilla almond beverage, and (30 g) of cheddar cheese or plant-based cheese. Blood glucose, insulin, and appetite were measured at baseline, post-treatment, and following a fixed-size pasta meal (post-meal) within 15-30 min. In experiment two, C-peptide, GLP-1, and ghrelin were measured. Greek yogurt and cheddar cheese lowered post-meal blood glucose more than their plant-based alternatives (p <0.01) and post-treatment blood glucose was higher following almond beverage than cheddar cheese and plant-based cheese (p <0.01). In experiment 1, post-treatment insulin was higher after Greek yogurt than cheddar cheese and plant-based cheese and all treatments post-meal (p <0.02). Post-meal appetite was lower after plant-based yogurt than cheddar cheese and plant-based cheese (p <0.01). In experiment 2, post-treatment insulin was higher after almond beverage compared to all treatments (p <0.01) and post-meal GLP-1 was higher after milk than almond beverage (p =0.03). We conclude that the physiological functionality of plant-based alternatives as measured by blood glucose, insulin, C-peptide, and GLP-1 did not replicate the metabolic functions of dairy products.

Methyl donor micronutrients, hypothalamic development and programming for metabolic disease.

Nutriture in utero is essential for fetal brain development through the regulation of neural stem cell proliferation, differentiation, and apoptosis, and has a long-lasting impact on risk of disease in offspring. This review examines the role of maternal methyl donor micronutrients in neuronal development and programming of physiological functions of the hypothalamus, with a focus on later-life metabolic outcomes. Although evidence is mainly derived from preclinical studies, recent research shows that methyl donor micronutrients (e.g., folic acid and choline) are critical for neuronal development of energy homeostatic pathways and the programming of characteristics of the metabolic syndrome in mothers and their children. Both folic acid and choline are active in one-carbon metabolism with their impact on epigenetic modification of gene expression. We conclude that an imbalance of folic acid and choline intake during gestation disrupts DNA methylation patterns affecting mechanisms of hypothalamic development, and thus elevates metabolic disease risk. Further investigation, including studies to determine translatability to humans, is required.

Caloric beverages consumed freely at meal-time add calories to an ad libitum meal.

The objective was to compare the effects of ad libitum consumption of commonly consumed meal-time beverages on energy and fluid intakes and post-meal average subjective appetite and blood glucose in healthy adults. In a randomized controlled design, 29 males and females consumed to satiation an ad libitum pizza meal with one of five beverages in unlimited amount including water (0 kcal), 1% milk (44 kcal/100 ml), regular cola (44 kcal/100 ml), orange juice (44 kcal/100 ml) and diet cola (0 kcal). Food and fluid intakes were measured at the meal. Average subjective appetite and blood glucose were measured before and for 2h after the meal. Although energy intake from pizza was similar among all beverage treatments, the amount of fluid consumed (g) varied among the beverages with intake of orange juice higher than regular and diet cola, but not different from water or milk. Meal-time ingestion of caloric beverages, milk, orange juice and regular cola, led to higher total meal-time energy intakes compared to either water or diet cola. Post-meal blood glucose area under the curve (AUC) was lower after milk than after meals with water, orange juice and regular cola and post-meal average subjective appetite AUC was lower after milk than after meals with water. Meal intakes of nutrients including protein, calcium, phosphorus, zinc, vitamins B12, A and D were higher at the meal with milk compared to the other beverages. Thus, caloric beverages consumed ad libitum during a meal add to total meal-time energy intake, but 1% milk favors a lower post-meal blood glucose and average subjective appetite score and adds to nutrient intake.

Micronutrients in High-Fat Diet Modify Insulin Resistance and Its Regulatory Genes in Adult Male Mice.

SCOPE: Obesity and insulin resistance (IR) are associated with epigenetic changes of gene expression. However, the relationship between micronutrients, epigenetic regulation of gene expression, and IR during development of diet-induced obesity has yet to be defined. Our objective is to describe the effect of micronutrient addition to diets on IR and its related genes during obesity development. METHODS AND RESULTS: Male C57BL/6J mice are fed a high-fat (HFD) or low-fat (LFD) diets with or without a multi-vitamin mineral mix (MVM) addition containing vitamins A, B1, B6, B12, and Zn, and Se for 9 weeks. Compared to LFD mice, HFD mice have higher body weight, IR, fasting glucose, insulin, C-peptide, leptin, and hepatic triglyceride concentrations, and dysregulated gene expression in liver, muscle, pancreas, and fat tissues (p < 0.05). The addition of MVM reduces these HFD-induced effects. HFD downregulates 27 genes associated with insulin regulation and adipose tissue function across all tissues by an average of 47% and upregulates five genes by 230% (p < 0.001). Adding MVM downregulates five genes and upregulates one in HFD-fed mice. Both HFD and MVM alter one-carbon metabolites. CONCLUSION: Addition of micronutrients to the HFD decreases IR and modifies associated gene expression in obese and lean mice.

The use of low-calorie sweeteners by adults: impact on weight management.

The application of low-calorie sweeteners (LCS) in foods and beverages has increased over the past 35 y. At the same time, many characteristics of the American diet have changed, including variations in fat and carbohydrate content and composition, increased nutrient additions, and new dietary patterns due to changing lifestyles and attitudes toward food and the changing cost of food. During this same time period, the prevalence of overweight and obesity has increased from ~30 to 70% of adults in the United States. Clearly, these trends lead to a variety of hypotheses and efforts to explain the role of LCS in this association. The aim of this review is to gain clarity on the role of LCS in weight management and their impact on diet quality. In addition, because the majority of studies aimed at identifying associations between LCS and these outcomes are based on observational data, the pitfalls in designing and evaluating data from observational studies are also discussed. We conclude that there is no evidence that LCS can be claimed to be a cause of higher body weights in adults. Similarly, evidence supporting a role for LCS in weight management is lacking. Due to the confounders in most observational studies, randomized controlled trials are needed to advance understanding.