Isocaloric Intake Research Articles

FRI391 A Metabolomics Analysis Of Moderate Energy Restriction In Healthy Lean Women

Abstract Disclosure: L. Levine: None. K. Overdahl: None. S. Molsberry: None. S. Shekhar: None. A. Jarmusch: None. J.E. Hall: None. Introduction: Metabolomics can elucidate metabolic responses to dietary interventions and has been used in energy restriction studies, primarily in overweight/obese men and women. Although the menstrual cycle affects the metabolome, most dietary intervention studies have not controlled for menstrual cycle phase, nor have they examined the effect of moderate, short-term energy restriction on the metabolic profile of healthy, normal-weight women. Objective: To characterize metabolomic changes in response to a 55% reduction in dietary intake during the early follicular phase of the menstrual cycle. Methods: Study participants included 19 healthy women aged 18-28 years who were of normal BMI and fitness, not restricting nutritional intake and with biochemically confirmed regular ovulatory menstrual cycles. Participants underwent 5-day dietary interventions in the early follicular phase of two menstrual cycles. The first diet consisted of neutral energy availability (NEA; 45 kcal/kg*lean body mass/day) and the second of deficient energy availability (DEA; 20 kcal/kg*lean body mass/day) without changes in macronutrient composition. On the 5th day of each diet, serum was collected at ∼8am following an overnight fast, and immediately before and every 10 minutes for 60 minutes after the consumption of a standardized isocaloric snack at ∼4pm. Untargeted metabolomic profiling was performed using ultra-high performance liquid chromatography/mass spectroscopy. Univariate statistics were performed on all metabolites, using t-tests with Benjamini-Hochberg false discovery rate adjusted p-values to account for multiple testing (p-value ≤ 0.05). Log2-fold change was calculated for all chemical features. Results: At baseline, participants had a weight of 61.9±2.75 kg (mean±SEM), BMI of 23.96±0.69 kg/m2 (mean±SEM), and percent body fat of 28.01±1.61% (mean±SEM). DEA resulted in a 1.25±0.13 kg (mean±SEM) decrease in body weight (p<0.01). While few statistically significant differences were present between fasted NEA and DEA samples, numerous chemical features were significantly upregulated in DEA post-snack samples. Specifically, 11 carnitines, 7 phosphocholines, and 8 fatty acids or fatty acid intermediates exhibited a log2fold change of 0.5 or higher (>40% increase; p ≤ 0.05) in post-snack DEA samples compared to NEA samples. Conclusion: These results strongly suggest that following isocaloric intake, moderate energy restriction in normal-weight, healthy women is associated not only with the use of fat as an energy source, but also with cellular membrane breakdown. Presentation: Friday, June 16, 2023

Additional Resistant Starch from One Potato Side Dish per Day Alters the Gut Microbiota but Not Fecal Short-Chain Fatty Acid Concentrations.

The composition of the gut microbiota and their metabolites are associated with cardiometabolic health and disease risk. Intake of dietary fibers, including resistant starch (RS), has been shown to favorably affect the health of the gut microbiome. The aim of this research was to measure changes in the gut microbiota and fecal short-chain fatty acids as part of a randomized, crossover supplemental feeding study. Fifty participants (68% female, aged 40 ± 13 years, BMI 24.5 ± 3.6 kg/m2) completed this study. Potato dishes (POT) contained more RS than refined grain dishes (REF) (POT: 1.31% wet basis (95% CI: 0.94, 1.71); REF: 0.73% wet basis (95% CI: 0.34, 1.14); p = 0.03). Overall, potato dish consumption decreased alpha diversity, but beta diversity was not impacted. Potato dish consumption was found to increase the abundance of Hungatella xylanolytica, as well as that of the butyrate producing Roseburia faecis, though fecal butyrate levels were unchanged. Intake of one potato-based side dish per day resulted in modest changes in gut microbiota composition and diversity, compared to isocaloric intake of refined grains in healthy adults. Studies examining foods naturally higher in RS are needed to understand microbiota changes in response to dietary intake of RS and associated health effects.

Jejunal mucosa proteomics unravel metabolic adaptive processes to mild chronic heat stress in dairy cows

Climate change affects the duration and intensity of heat waves during summer months and jeopardizes animal health and welfare. High ambient temperatures cause heat stress in dairy cows resulting in a reduction of milk yield, feed intake, and alterations in gut barrier function. The objectives of this study were to investigate the mucosal amino acid, glucose and lactate metabolism, as well as the proteomic response of the small intestine in heat stressed (HS) Holstein dairy cows. Cows of the HS group (n = 5) were exposed for 4 days to 28 °C (THI = 76) in a climate chamber. Percentage decrease in daily ad libitum intake of HS cows was calculated to provide isocaloric energy intake to pair-fed control cows kept at 15 °C (THI = 60) for 4 days. The metabolite, mRNA and proteomic analyses revealed that HS induced incorrect protein folding, cellular destabilization, increased proteolytic degradation and protein kinase inhibitor activity, reduced glycolysis, and activation of NF-κB signaling, uronate cycling, pentose phosphate pathway, fatty acid and amino acid catabolism, mitochondrial respiration, ATPase activity and the antioxidative defence system. Our results highlight adaptive metabolic and immune mechanisms attempting to maintain the biological function in the small intestine of heat-stressed dairy cows.

The Effects of 2-Week 16:8-Hour Intermittent Fasting on Body Composition: Case Study

Intermittent fasting is a form of nutrition dieting in which an individual is allotted an amount of time to consume food throughout the day. Intermittent fasting has been shown to acutely change an individual’s body composition as an effective intervention. The purpose of this research was to determine the effects of 2-week 16:8-hour intermittent fasting on body composition. Participants ages ranged from 21-24 years, with a total of four (males: 3, female: 1), all college aged students. All participants were involved in regular exercise but had no previous experience of intermittent fasting. Each participant consumed their normal dietary intake till day of pre-intervention testing but were told to be in a fasted state before undergoing baseline testing. A glucometer was used to evaluate fasted blood glucose. Blood pressure was determined by using a manual cuff. A stadiometer was used to measure participants height and weight. A flexible tape measure was used to determine the waist and hip circumferences, which were used to determine the waist to hip ratio. Bioelectrical Impedance Analysis and Skinfold Calipers were used to determine body composition. Participants maintained normal physical activity levels, and logged volume and intensities in the MyFitnessPal application. Participants followed an isocaloric intake. Participants followed a 16:8 ratio, with a 16 hour fast and 8 hours allotted to consume food. Participants followed the intermittent fasting diet for a total of two weeks. Post-intervention was reevaluated at the two-week mark following intervention start. Data was presented as mean ± standard deviation and determined by descriptive analysis. Statistical significance was set at p < .05. The results may indicate that intermittent fasting with a 16:8 ratio can decrease body mass and resting blood pressure in young individuals. Due to COVID-19, limitations were placed allotting only 2 weeks for intervention and for smaller sample size. For future research, study would be increased to 8 weeks with an increased sample size up.

Time-restricted feeding improves the reproductive function of female mice via liver fibroblast growth factor 21.

BackgroundThere has been a significant increase, to epidemic levels, of obese and overweight women of reproductive age, causing impairments to reproductive health. Time‐restricted feeding (TRF) including isocaloric intake has shown to be preventive of obesity‐related disorders. However, its therapeutic ability to improve the reproductive function of female remains largely unknown.MethodsHere, we investigated the ability of TRF to improve the reproductive function in wild‐type and liver‐specific FGF21 knockout female mice. To study fertility, a continuous and a short‐term fertility test, gonadotropin releasing‐hormone (GnRH), and Kisspeptin test were performed. Immortalized GnRH neuron was used to examine the direct role of liver fibroblast growth factor 21 (FGF21) on GnRH secretion.ResultsWe found that TRF rescues female mice from bodyweight gain and glucose intolerance, as well as ovarian follicle loss and dysfunction of estrus cyclicity induced by high‐fat diet. Furthermore, the beneficial effects of the TRF regimen on the reproductive performance were also observed in mice fed both chow and high‐fat diet. However, those beneficial effects of TRF on metabolism and reproduction were absent in liver‐specific FGF21 knockout mice. In vitro, FGF21 directly acted on GnRH neurons to modulate GnRH secretion via extracellular regulated protein kinases (ERK1/2) pathway.ConclusionsOverall, time‐restricted feeding improves the reproductive function of female mice and liver FGF21 signaling plays a key role in GnRH neuron activity in female mice.

Influence of dietary fructose supplementation on visceral organ mass, carbohydrase activity, and mRNA expression of genes involved in small intestinal carbohydrate assimilation in neonatal calves

The hypothesis of this experiment was that dietary fructose would influence visceral organ mass, carbohydrase activity, and mRNA expression of carbohydrases and nutrient transporters in the small intestine in neonatal calves. Therefore, our objective was to use the neonatal calf as a model to evaluate the effects of postruminal fructose supply on small intestinal carbohydrate assimilation. Ten calves (<7 d of age; 41.2 ± 1.46 kg of body weight) were fed milk replacer at 2.0% of body weight daily (816 ± 90.5 g/d; 272 ± 30.1 g/L; dry-matter basis) in 2 equal portions and assigned to the following dietary treatment groups: (1) milk replacer (control; n = 6) or (2) milk replacer + 2.2 g of fructose/kg of body weight (fructose; n = 4). Calves were fed dietary treatments for 28 d, with jugular blood sampled every 7 d before and after the morning feeding. Calves were slaughtered, and visceral weights were recorded. Postruminal carbohydrase activities were assayed. Quantitative real-time PCR was conducted for small intestinal mRNA expression of nutrient transporters [solute carrier family 2 member 5 (GLUT5), solute carrier family 2 member 2 (GLUT2), and solute carrier family 5 member 1 (SGLT1)], carbohydrases (lactase, maltase-glucoamylase, and sucrase-isomaltase), and ketohexokinase (KHK). Data were analyzed using MIXED procedures in SAS version 9.4 (SAS Institute Inc, Cary, NC). Dietary fructose supplementation decreased serum glucose concentration. Small intestinal mass was greater in calves supplemented with fructose. Dietary fructose supplementation did not influence pancreatic α-amylase, small intestinal isomaltase, or maltase activities. Sucrase activity was undetected in the small intestine. Dietary fructose supplementation increased small intestinal glucoamylase activity per gram of tissue by 30% and increased maltase-glucoamylase mRNA expression by 6.8-fold. Dietary fructose supplementation did not influence mRNA expression of GLUT5, SGLT1, GLUT2, or KHK. Dietary fructose supplementation increased small intestinal lactase mRNA expression by 3.1-fold. Sucrase-isomaltase mRNA expression in the small intestine decreased 5.1-fold with dietary fructose supplementation. Dietary fructose supplementation does not induce sucrase activity in neonatal calves; however, sucrase-isomaltase may be transcriptionally regulated by dietary fructose in neonatal calves. More research is needed to compare glucose and fructose at isocaloric intakes to examine effects of dietary fructose at equal metabolizable energy intake.

Behavioral and cortisol responses to feeding frequency in pregnant sows under isocaloric intake.

The study focused on behavioral and cortisol responses to feeding frequency in pregnant sows under isocaloric intake. Twenty-four sows [(Landrace × Yorkshire); BW 216.70 ± 3.98 kg; parity 3.04 ± 0.53] were balanced for parity and randomly assigned to 1 of 3 feeding frequency regimes. Sows were fed corn–soybean meal-based diet 1× [0730 (Control), T1], 2× [half ration at 0730 and 1530 hours, T2], or 3× [one-third portion at 0730, 1130, and 1530 hours, T3] from days 30 to 60 of gestation. Sows received 7055 kcal ME/d during gestation from 2.21 kg of diet formulated to contain SID Lys/ME of 1.71 g/Mcal. Saliva samples were collected every 2 hr from 0630 to 1830 hours on day 52 and assayed for cortisol using ELISA procedure. Behavior data were collected for 7 d from day 53 of gestation by affixing a remote insights ear tag to each sow. Each sow had 120,960 data points categorized into: “Active”, “Feed,” or “Dormant”. Because of housing constraint, all sows were housed in individual stalls in the same room presenting a potential limitation of the study. The data were analyzed using PROC MIXED and GLIMMIX procedures of SAS 9.4 for cortisol and behavior count data, respectively. Sow was the experimental unit. The area under the curve (AUC) is quantitative evaluation of response as threshold varies over all possible values. The T2 sows had reduced 12-hr cortisol AUC compared with control sows (P = 0.024) and T3 sows (P = 0.004), respectively. The T2 sows had lower 3 hr (P = 0.039) and 5 hr (P = 0.015) postfeeding cortisol AUC compared with control sows. Feed anticipatory activity (FAA), 24-hr total activity, and feeding activities (eating and/or sham chewing) were reduced for T2 sows relative to the control and T3 sows (P < 0.01). Consequently, T2 sows had lower 24-hr total activity (P < 0.001) and feeding activities (P < 0.001) AUC compared with both the control and T3 sows, respectively. The T3 sows had greater FAA (P < 0.001) and 24-hr total activity AUC (P = 0.010) compared with control sows. Our data although inconclusive due to small sample size, twice daily feeding appears to be the threshold that reduces sows’ total activity AUC, feeding activity AUC, and activation of hypothalamic–pituitary–adrenal axis, reduced hunger, and exhibit potential to improve sow welfare in relation to once and thrice daily feeding regimes under isocaloric intake per kilogram live metabolic weight.

Improvement in Protein Delivery for Critically Ill Patients Requiring High-Dose Propofol Therapy and Enteral Nutrition.

Patients with traumatic brain (TBI) injury often require a high dosage of propofol, which can provide an excessive caloric intake. We evaluated our strategy of using liquid protein supplement boluses concurrently with high protein-containing enteral nutrition (EN) formulas and formula rate reduction to avoid caloric overfeeding and inadequate protein intake. Adult patients (aged >17 years) with TBI admitted to the trauma intensive care unit (TICU) who received concurrent propofol and EN were retrospectively reviewed. Caloric intakes from propofol and EN were obtained. Actual protein intake was compared with projected protein intakes from high protein content and standard protein content enteral formulas when given at an isocaloric intake. Fifty-one patients were enrolled. Average caloric intake from propofol was 356 ± 243 kcal/d or 5 ± 3 kcal/kg/d (range, <1-15 kcal/kg/d). Daily EN caloric intake ranged from 7 ± 4 kcal/kg/d (day 2) to 16 ± 9 kcal/kg/d (day 5; P < .001). Average protein intake ranged from 0.6 ± 0.4 g/kg/d (day 2) to 1.5 ± 0.7 g/kg/d (day 5; P < .001). The modified EN strategy resulted in daily delivery of 24%-38% more protein than an isocaloric regimen with a high protein-content formula and twice as much protein than the standard protein-content formula (P < .001). The strategy of providing an EN regimen comprised liquid protein boluses, and high and very high protein-containing EN formulas at a reduced rate improved protein delivery without caloric overfeeding.

Identification of Milk and Cheese Intake Biomarkers in Healthy Adults Reveals High Interindividual Variability of Lewis System–Related Oligosaccharides

ABSTRACTBackgroundThe use of biomarkers of food intake (BFIs) in blood and urine has shown great promise for assessing dietary intake and complementing traditional dietary assessment tools whose use is prone to misreporting.ObjectiveUntargeted LC-MS metabolomics was applied to identify candidate BFIs for assessing the intake of milk and cheese and to explore the metabolic response to the ingestion of these foods.MethodsA randomized controlled crossover study was conducted in healthy adults [5 women, 6 men; age: 23.6 ± 5.0 y; BMI (kg/m2): 22.1 ± 1.7]. After a single isocaloric intake of milk (600 mL), cheese (100 g), or soy-based drink (600 mL), serum and urine samples were collected postprandially up to 6 h and after fasting after 24 h. Untargeted metabolomics was conducted using LC-MS. Discriminant metabolites were selected in serum by multivariate statistical analysis, and their mass distribution and postprandial kinetics were compared.ResultsSerum metabolites discriminant for cheese intake had a significantly lower mass distribution than metabolites characterizing milk intake (P = 4.1 × 10−4). Candidate BFIs for milk or cheese included saccharides, a hydroxy acid, amino acids, amino acid derivatives, and dipeptides. Two serum oligosaccharides, blood group H disaccharide (BGH) and Lewis A trisaccharide (LeA), specifically reflected milk intake but with high interindividual variability. The 2 oligosaccharides showed related but opposing trends: subjects showing an increase in either oligosaccharide did not show any increase in the other oligosaccharide. This result was confirmed in urine.ConclusionsNew candidate BFIs for milk or cheese could be identified in healthy adults, most of which were related to protein metabolism. The increase in serum of LeA and BGH after cow-milk intake in adults calls for further investigations considering the beneficial health effects on newborns of such oligosaccharides in maternal milk. The trial is registered at clinicaltrials.gov as NCT02705560.

Effect of feeding frequency and sow parity based on isocaloric intake during gestation on sow performance.

The aim of this study was to investigate the effect of feeding frequency and sow parity based on same rate of maintenance energy intake during gestation on sow performance. One hundred and seventy-seven sows [Topigs Norsvin 70, Landrace × Large White, Topigs Norsvin USA, Burnsville, MN; parity 3.80 ± 0.16; initial BW = 211.34 ± 3.37 kg; backfat (BF) 13.57 ± 0.54 mm] were blocked by parity, balanced for BW, and randomly assigned to 1 of 3 treatments in a randomized complete block design. Treatments included sows fed corn-soybean meal-based diet 1× daily at 0730 h (control, T1), 2× daily [half ration at 0730 and 1530 h (T2)], or 3× daily [a third portion at 0730, 1130, and 1530 h (T3)], with daily feed quantity kept at 1.25 × maintenance energy intake [100 × (BW)0.75] kcal ME/d. Treatments were imposed from day 30 of gestation. Sows received on average 6,921, 7,129, and 7,399 kcal ME/d from days 30 to 60, days 61 to 90, days 91 to 109 of gestation, respectively. Feeding frequency during gestation had no effect on lactation ADFI (P > 0.10). Sows fed 3× daily during gestation had improved lactation G:F (P = 0.040) compared with sows fed 2× but similar to control sows (P = 0.338). Treatment did not alter BW or BW variations during gestation, lactation, or from days 30 to wean (P > 0.10). Sows fed 2× daily had tendency to gain BF from day 30 to day 109 of gestation (P = 0.053) but tended to lose BF during lactation (P = 0.091) relative to the control sows. Feeding frequency (2× and 3× daily) tended to increase the number of piglets weaned by 0.40 (P = 0.056) and 0.53 (P = 0.098) piglets, respectively, compared with control sows. Sows fed 2× daily during gestation had reduced number of stillborn relative to control sows (P = 0.035). From day 30 to wean, gilts had propensity to lose BF relative to P1+ (P = 0.094), but lost BF compared with P3+ and P6+ sows (P = 0.003). Parity P6+ sows had highest percentage of both 72 h and preweaning piglet mortality than P0, P1+, and P3+ sows (P < 0.05). In conclusion, parity (P6+) had greater lactation BW gain but higher mortalities relative to lower parity sows. Sows fed 2× daily tended to gain BF from days 30 to 109 of gestation and had reduced number of stillborn relative to control sows. It appears that increasing gestation sow feeding frequency from 1× daily to 2× and 3× daily may reduce the number of stillborn and increase litter size at weaning although most of the reproductive traits were not affected.

The Effect of Glycomacropeptide versus Amino Acids on Phenylalanine and Tyrosine Variability over 24 Hours in Children with PKU: A Randomized Controlled Trial.

Introduction: In phenylketonuria (PKU), evidence suggests that casein glycomacropeptide supplemented with rate-limiting amino acids (CGMP-AA) is associated with better protein utilisation and less blood phenylalanine (Phe) variability. Aim: To study the impact of CGMP-AA on blood Phe variability using 3 different dietary regimens in children with PKU. Methods: This was a 6-week randomised controlled cross-over study comparing CGMP-AA vs. Phe-free l-amino acids (l-AA) assessing blood Phe and tyrosine (Tyr) variability over 24 h in 19 children (7 boys) with PKU, with a median age of 10 years (6–16). Subjects were randomised to 3 dietary regimens: (1) R1, CGMP-AA and usual dietary Phe (CGMP + Phe); (2) R2, CGMP-AA − Phe content of CGMP-AA from usual diet (CGMP − Phe); and (3) R3, l-AA and usual dietary Phe. Each regimen was administered for 14 days. Over the last 48 h on days 13 and 14, blood spots were collected every 4 h at 08 h, 12 h, 16 h, 20 h, 24 h, and 04 h. Isocaloric intake and the same meal plan and protein substitute dosage at standardised times were maintained when blood spots were collected. Results: Eighteen children completed the study. Median Phe concentrations over 24 h for each group were (range) R1, 290 (30–580), R2, 220 (10–670), R3, 165 (10–640) μmol/L. R1 vs. R2 and R1 vs. R3 p < 0.0001; R2 vs. R3 p = 0.0009. There was a significant difference in median Phe at each time point between R1 vs. R2, p = 0.0027 and R1 vs. R3, p < 0.0001, but not between any time points for R2 vs. R3. Tyr was significantly higher in both R1 and R2 [70 (20–240 μmol/L] compared to R3 [60 (10–200) μmol/L]. In children < 12 years, blood Phe remained in the target range (120–360 μmol/L), over 24 h, for 75% of the time in R1, 72% in R2 and 64% in R3; for children aged ≥ 12 years, blood Phe was in target range (120–600 μmol/L) in R1 and R2 for 100% of the time, but 64% in R3. Conclusions: The residual Phe in CGMP-AA increased blood Phe concentration in children. CGMP-AA appears to give less blood Phe variability compared to l-AA, but this effect may be masked by the increased blood Phe concentrations associated with its Phe contribution. Reducing dietary Phe intake to compensate for CGMP-AA Phe content may help.

Impact of Free-Choice Diets High in Fat and Different Sugars on Metabolic Outcome and Anxiety-Like Behavior in Rats.

ObjectiveRats were exposed to free‐choice diets (fat plus one of two different sugar solutions, glucose or sucrose), and the metabolic consequences and impact on locomotor activity and anxiety‐like behavior were explored.MethodsFor 3 weeks, 7‐week‐old male rats were offered either chow only or free‐choice high‐fat diets differing in their added sugar: no sugar, sucrose, or glucose. In a second experiment, after 2 weeks on the diets, rats were switched from high sucrose to high glucose for two additional weeks. Metabolic end points included body weight, food intake, food choice, glycemic control, metabolic hormones, fat pad weight, brown adipose tissue weight, and gene expression. Behavioral analysis included locomotor and anxiety‐like activity in the open field and elevated plus maze.ResultsBoth sugar diets enhanced adiposity and induced hyperphagia, favoring unhealthier dietary selection above that of the control diets (chow or free‐choice high‐fat with no sugar). Despite isocaloric intake in the sugar‐containing diets, offering glucose instead of sucrose was associated with improved insulin sensitivity. The sugar‐containing diets reduced activity (but with movements of increased velocity) and induced an anxiety‐like phenotype.ConclusionsAlthough free‐choice diets negatively impacted on metabolism and anxiety‐like behavior, replacing sucrose with glucose improved insulin sensitivity and may therefore be better for health.

Naringenin Supplementation to a Chow Diet Enhances Energy Expenditure and Fatty Acid Oxidation, and Reduces Adiposity in Ldlr-/- Mice with Isocaloric Food Intake

Naringenin Supplementation to a Chow Diet Enhances Energy Expenditure and Fatty Acid Oxidation, and Reduces Adiposity in Ldlr-/- Mice with Isocaloric Food Intake

Effect of parenteral glutamine supplementation combined with enteral nutrition on Hsp90 expression and Peyer's patch apoptosis in severely burned rats.

The aim of this study was to investigate the effects of parenteral glutamine (GLN) supplementation combined with enteral nutrition (EN) on heat shock protein (Hsp) 90 expression and Peyer's patch (PP) apoptosis in severely burned rats. Male Sprague-Dawley (SD) rats were randomly assigned to four groups: Sham burn+EN+GLN-free amino acid (AA; n=10), sham burn+EN+GLN (n=10), burn+EN+AA (n=10), and burn+EN+GLN (n=10). Two hours after a 30% total body surface area (TBSA), full-thickness scald burn injury on the back, burned rats in two of the experimental groups (burn+EN+AA and burn+EN+GLN groups) were fed with a conventional EN solution by oral gavage for 7d. Simultaneously, rats in the burn+EN+GLN group were given 0.35g GLN/kg body weight/d once via a tail vein injection for 7d and rats in the burn+EN+AA group were administered isocaloric/isonitrogenous GLN-free amino acid solution (Tyrosine) for comparison. Rats in two sham burn control groups (sham burn+EN+AA and sham burn+EN+GLN groups) were treated in the same manner except for the burn injury. All rats in the four groups were given 175kcal/kg body wt/d. There was isonitrogenous, isovolumic, and isocaloric intake among the four groups. At the end of the seventh day after completion of the nutritional program, all rats were anesthetized and samples were collected for further analysis. PP apoptosis was measured by terminal deoxyuridine nick-end labeling (TUNEL). The expression of Hsp90 in PPs was analyzed by western blotting. Caspase-3 activity of PPs was also assessed. Levels of proinflammatory cytokines of gut tissues were evaluated by enzyme-linked immunosorbent assay (ELISA). The intestinal immunoglobulin A (IgA) content was also determined by ELISA. The results revealed that intestinal IgA content in rats of the burn+EN+GLN group were significantly increased compared with those in the burn+EN+AA group (P<0.05). The expression of Hsp90 of PPs in rats in the burn+EN+GLN group was significantly upregulated compared with those in the burn+EN+AA group (P<0.05). On the other hand, levels of proinflammatory cytokines of gut tissues, caspase-3 activity, and the number of TUNEL-stained cells of PPs in rats of the burn+EN+GLN group were markedly decreased compared with those of the burn+EN+AA group (P<0.05). The results of this study show that parenteral glutamine supplementation combined with EN may upregulate the expression of Hsp90, reduce caspase-3 activity, lessen the release of proinflammatory cytokines, attenuate PP apoptosis, and improve intestinal IgA response in burned rats. Clinically, therapeutic efforts to improve intestinal immunity may contribute to a favorable outcome in severely burned patients.

Effect of parenteral glutamine supplementation combined with enteral nutrition on Hsp90 expression and lymphoid organ apoptosis in severely burned rats

ObjectiveThe aim of this study is to investigate the effects of parenteral glutamine(GLN) supplementation combined with enteral nutrition (EN) on heat shock protein 90(Hsp90) expression, apoptosis of lymphoid organs and circulating lymphocytes, immunological function and survival in severely burned rats. MethodsMale SD rats were randomly assigned into 4 groups: a sham burn+EN+GLN-free amino acid (AA) group (n=10), a sham burn+EN+GLN group (n=10), a burn+EN+AA group (n=10), and a burn +EN +GLN group (n=10). Two hours after a 30% total body surface area (TBSA), full-thickness scald burn injury on the back was made, the burned rats in two experimental groups (the burn+EN+AA group and the burn+EN +GLN group) were fed with a conventional enteral nutrition solution by oral gavage for 7 days. Simultaneously, the rats in the burn+EN+GLN group were given 0.35g GLN/kg body weight/day once via a tail vein injection for 7 days, whereas those in the burn+EN+AA group were administered isocaloric/isonitrogenous GLN-free amino acid solution (Tyrosine) for comparison. The rats in two sham burn control groups (the sham burn+EN+AA group and the sham burn+EN +GLN group) were treated in the same procedure as above, except for burn injury. All rats in each of the four groups were given 175kcal/kg body wt/day. There was isonitrogenous, isovolumic and isocaloric intake among four groups. At the end of the 7th day after nutritional programme were finished, all rats were anesthetized and samples were collected for further analysis. Serum immunoglobulin quantification was conducted by ELISA. Circulating lymphocyte numbers were counted by Coulter LH-750 Analyzer. The percentages and apoptotic ratio of CD4 and CD8T lymphocytes in circulation were determined by flow cytometry (FCM). The neutrophil phagocytosis index (NPI) was examined. The GLN concentrations in plasma, thymus, spleen and skeletal muscle were measured by high performance liquid chromatography (HPLC). The organ index evaluation and TUNEL analysis of thymus and spleen were carried out. The expression of Hsp90 in thymus and spleen was analyzed by western blotting. Moreover, the survival in burned rats was observed. ResultsThe results revealed that parenteral GLN supplementation combined with EN significantly increased the GLN concentrations of plasma and tissues, the serum immunoglobulin content, the circulating lymphocyte number, the CD4/CD8 ratio, the indexes of thymus and spleen, NPI and survival as compared with the burn+EN+AA group (p<0.05). The expression of Hsp90 in thymus and spleen in the burn+EN+GLN group was significantly up-regulated as compared with the burn+EN+AA group (p<0.05). The apoptosis in circulating CD4 and CD8 lymphocytes, thymus and spleen in the burn+EN+GLN group was significantly decreased as compared with the burn+EN+AA group (p<0.05). ConclusionThe results of this study show that parenteral GLN supplementation combined with EN may increase the GLN concentrations of plasma and tissues, up-regulate the expression of Hsp90, attenuate apoptosis in lymphoid organ and circulating lymphocyte, enhance the immunological function and improve survival in severely burned rats. Clinically, therapeutic efforts at the modulation of the immune dysfunction may contribute to a favorable outcome in severely burned patients.

High Intrinsic Aerobic Capacity Protects against Ethanol-Induced Hepatic Injury and Metabolic Dysfunction: Study Using High Capacity Runner Rat Model.

Rats artificially selected over several generations for high intrinsic endurance/aerobic capacity resulting in high capacity runners (HCR) has been developed to study the links between high aerobic fitness and protection from metabolic diseases (Wisloff et al., Science, 2005). We have previously shown that the HCR strain have elevated hepatic mitochondrial content and oxidative capacity. In this study, we tested if the elevated hepatic mitochondrial content in the HCR rat would provide “metabolic protection” from chronic ethanol-induced hepatic steatosis and injury. The Leiber-Decarli liquid diet with ethanol (7% v/v; HCR-E) and without (HCR-C) was given to HCR rats (n = 8 per group) from 14 to 20 weeks of age that were weight matched and pair-fed to assure isocaloric intake. Hepatic triglyceride (TG) content and macro- and microvesicular steatosis were significantly greater in HCR-E compared with HCR-C (p < 0.05). In addition, hepatic superoxide dismutase activity and glutathione levels were significantly (p < 0.05) reduced in the HCR-E rats. This hepatic phenotype also was associated with reduced total hepatic fatty acid oxidation (p = 0.03) and β-hydroxyacyl-CoA dehydrogenase activity (p = 0.01), and reductions in microsomal triglyceride transfer protein and apoB-100 protein content (p = 0.01) in HCR-E animals. However, despite these documented hepatic alterations, ethanol ingestion failed to induce significant hepatic liver injury, including no changes in hepatic inflammation, or serum alanine amino transferase (ALTs), free fatty acids (FFAs), triglycerides (TGs), insulin, or glucose. High intrinsic aerobic fitness did not reduce ethanol-induced hepatic steatosis, but protected against ethanol-induced hepatic injury and systemic metabolic dysfunction in a high aerobic capacity rat model.

Potential link between excess added sugar intake and ectopic fat: a systematic review of randomized controlled trials.

The effect of added sugar intake on ectopic fat accumulation is a subject of debate. A systematic review and meta-analysis of randomized controlled trials (RCTs) was conducted to examine the potential effect of added sugar intake on ectopic fat depots. MEDLINE, CAB Abstracts, CAB Global Health, and EBM (Evidence-Based Medicine) Reviews - Cochrane Central Register of Controlled Trials databases were searched for studies published from 1973 to September 2014. RCTs with a minimum of 6 days' duration of added sugar exposure in the intervention group were selected. The dosage of added sugar intake as a percentage of total energy was extracted or calculated. Means and standard deviations of pre- and post-test measurements or changes in ectopic fat depots were collected. Fourteen RCTs were included. Most of the studies had a medium to high risk of bias. Meta-analysis showed that, compared with eucaloric controls, subjects who consumed added sugar under hypercaloric conditions likely increased ectopic fat, particularly in the liver (pooled standardized mean difference = 0.9 [95%CI, 0.6-1.2], n = 6) and muscles (pooled SMD = 0.6 [95%CI, 0.2-1.0], n = 4). No significant difference was observed in liver fat, visceral adipose tissue, or muscle fat when isocaloric intakes of different sources of added sugars were compared. Data from a limited number of RCTs suggest that excess added sugar intake under hypercaloric diet conditions likely increases ectopic fat depots, particularly in the liver and in muscle fat. There are insufficient data to compare the effect of different sources of added sugars on ectopic fat deposition or to compare intake of added sugar with intakes of other macronutrients. Future well-designed RCTs with sufficient power and duration are needed to address the role of sugars on ectopic fat deposition.

Time-fixed feeding prevents obesity induced by chronic advances of light/dark cycles in mouse models of jet-lag/shift work

Recent findings have uncovered intimate relationships between circadian clocks and energy metabolism. Epidemiological studies have shown that the frequency of obesity and metabolic disorders increases among shift-workers. Here we found that a chronic shift in light/dark (LD) cycles comprising an advance of six hours twice weekly, induced obesity in mice. Under such conditions that imitate jet lag/shift work, body weight and glucose intolerance increased, more fat accumulated in white adipose tissues and the expression profiles of metabolic genes changed in the liver compared with normal LD conditions. Mice fed at a fixed 12 h under the LD shift notably did not develop symptoms of obesity despite isocaloric intake. These results suggest that jet lag/shift work induces obesity as a result of fluctuating feeding times and it can be prevented by fixing meal times. This rodent model of obesity might serve as a useful tool for understanding why shift work induces metabolic disorders.

Nutrient-adjusted high-fat diet is associated with absence of periepididymal adipose tissue inflammation: is there a link with adequate micronutrient levels?

The aim of this study was to investigate the real impact of dietary lipids on metabolic and inflammatory response in rat white adipose tissue. Male healthy Wistar rats were fed ad libitum with a control diet (CON, n=12) or with an adjusted high-fat diet (HFD, n=12) for 12 weeks. Oral glucose and insulin tolerance tests were performed during the last week of the protocol. Plasma fatty acid, lipid profile, body adiposity, and carcass chemical composition were analyzed. Plasma concentration of leptin, adiponectin, C-reactive protein (CRP), TNF-α, IL-6, and monocyte chemotactic protein (MCP-1) was measured. Periepididymal adipose tissue was employed to evaluate TNF-α, MCP-1, and adiponectin gene expression as well as NF-κB pathway and AKT proteins. Isocaloric intake of the adjusted HFD did not induce hyperphagia, but promoted an increase in periepididymal (HFD = 2.94 ± 0.77 vs. CON = 1.99 ± 0.26 g/100 g body weight, p = 0.01) and retroperitoneal adiposity (HFD = 3.11 ± 0.81 vs. CON = 2.08 ± 0.39 g/100 g body weight, p = 0.01) and total body lipid content (HFD = 105.3 ± 20.8 vs. CON = 80.5 ± 7.6 g carcass, p = 0.03). Compared with control rats, HFD rats developed glucose intolerance (p=0.01), dyslipidemia (p = 0.02) and exhibited higher C-reactive protein levels in response to the HFD (HFD = 1002 ± 168 vs. CON = 611 ± 260 ng/mL, p = 0.01). The adjusted HFD did not affect adipokine gene expression or proteins involved in inflammatory signaling, but decreased AKT phosphorylation after insulin stimulation in periepididymal adipose tissue (p = 0.01). In this study, nutrient-adjusted HFD did not induce periepididymal adipose tissue inflammation in rats, suggesting that the composition of HFD differently modulates inflammation in rats, and adequate micronutrient levels may also influence inflammatory pathways.

Three Dissimilar High Fat Diets Differentially Regulate Lipid and Glucose Metabolism in Obesity‐Resistant Slc:Wistar/ST Rats

Epidemiologic and ecologic studies suggest that dietary fat plays an important role in the development of obesity. Certain Wistar rat strains do not become obese when fed high-fat diets unlike others. In a preliminary study, we confirmed that Slc:Wistar/ST rats did not become obese when fed high-fat diets. The mechanisms governing the response of hepatic lipid-metabolizing enzymes to large quantities of dietary lipids consumed by obesity-resistant animals are unknown. The aim of the present study is to examine how obesity-resistant animals metabolize various types of high-fat diets and why they do not become obese. For this purpose, male Slc:Wistar/ST rats were fed a control low-fat diet (LS) or a high-fat diet containing fish oil (HF), soybean oil (HS), or lard (HL) for 4 weeks. We observed their phenotypes and determined lipid profiles in plasma and liver as well as mRNA expression levels in liver of genes related to lipid and glucose metabolism using DNA microarray and quantitative reverse transcriptase polymerase chain analyses. The body weights of all dietary groups were similar due to isocaloric intakes, whereas the weight of white adipose tissues in the LS group was significantly lower. The HF diet lowered plasma lipid levels by accelerated lipolysis in the peroxisomes and suppressed levels of very-low-density lipoprotein (VLDL) secretion. The HS diet promoted hepatic lipid accumulation by suppressed lipolysis in the peroxisomes and normal levels of VLDL secretion. The lipid profiles of rats fed the LS or HL diet were similar. The HL diet accelerated lipid and glucose metabolism.