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Item A period of 10 weeks of increased protein consumption does not alter faecal microbiota or volatile metabolites in healthy older men: a randomised controlled trial(Cambridge University Press on behalf of The Nutrition Society, 2020-07-02) Mitchell SM; McKenzie EJ; Mitchell CJ; Milan AM; Zeng N; D'Souza RF; Ramzan F; Sharma P; Rettedal E; Knowles SO; Roy NC; Sjödin A; Wagner K-H; O'Sullivan JM; Cameron-Smith DDiet has a major influence on the composition and metabolic output of the gut microbiome. Higher-protein diets are often recommended for older consumers; however, the effect of high-protein diets on the gut microbiota and faecal volatile organic compounds (VOC) of elderly participants is unknown. The purpose of the study was to establish if the faecal microbiota composition and VOC in older men are different after a diet containing the recommended dietary intake (RDA) of protein compared with a diet containing twice the RDA (2RDA). Healthy males (74⋅2 (sd 3⋅6) years; n 28) were randomised to consume the RDA of protein (0⋅8 g protein/kg body weight per d) or 2RDA, for 10 weeks. Dietary protein was provided via whole foods rather than supplementation or fortification. The diets were matched for dietary fibre from fruit and vegetables. Faecal samples were collected pre- and post-intervention for microbiota profiling by 16S ribosomal RNA amplicon sequencing and VOC analysis by head space/solid-phase microextraction/GC-MS. After correcting for multiple comparisons, no significant differences in the abundance of faecal microbiota or VOC associated with protein fermentation were evident between the RDA and 2RDA diets. Therefore, in the present study, a twofold difference in dietary protein intake did not alter gut microbiota or VOC indicative of altered protein fermentation.Item The Importance of Dietary Protein Quality in Mid- to High-Income Countries(Elsevier Inc on behalf of American Society for Nutrition, 2024-03) Moughan PJ; Fulgoni VL; Wolfe RRIn wealthy countries, the protein intake of adults is usually considered to be adequate, and considerations of protein quality are often deemed irrelevant. The objective was to examine dietary protein intakes of adults in developed countries in the context of dietary protein quality. An analysis of NHANES population data on actual protein intakes in the United States (a developed country) demonstrated that for a dietary Digestible Indispensable Amino Acid Score (DIAAS) of 100%, 11% of the adult (19-50 y) population had habitual protein intakes below the Estimated Average Requirement (EAR) and 22% below the Recommended Dietary Allowance. The percentage of the population with utilizable protein intakes potentially falling below the EAR increased as the assumed DIAAS declined. Analysis of the NHANES data and several other datasets also indicated that total protein intakes can be limiting or close to limiting for the elderly and some vegetarians and vegans. Here, lower dietary protein quality can potentially lead to inadequate utilizable protein intakes. For many people in specific physiological states (e.g., weight loss, endurance sports, resistance exercise) attempting to meet higher dietary protein targets often with accompanying lowered energy intakes, low dietary protein quality can lead to protein calories expressed as a proportion of total calories, falling outside what may be acceptable limits (maximum of 30% protein calories from total calories). In general, individuals within the adult population may be susceptible to macronutrient imbalance (whenever total protein intakes are high, daily energy intakes low) and for diets with lower protein quality (DIAAS <100%). Our analysis shows that dietary protein quality is relevant in mid- to high-income countries.Item Protein Intake at Twice the RDA in Older Men Increases Circulatory Concentrations of the Microbiome Metabolite Trimethylamine-N-Oxide (TMAO)(MDPI (Basel, Switzerland), 2019-09-12) Mitchell SM; Milan AM; Mitchell CJ; Gillies NA; D'Souza RF; Zeng N; Ramzan F; Sharma P; Knowles SO; Roy NC; Sjödin A; Wagner K-H; Zeisel SH; Cameron-Smith DHigher dietary protein intake is increasingly recommended for the elderly; however, high protein diets have also been linked to increased cardiovascular disease (CVD) risk. Trimethylamine-N-oxide (TMAO) is a bacterial metabolite derived from choline and carnitine abundant from animal protein-rich foods. TMAO may be a novel biomarker for heightened CVD risk. The purpose of this study was to assess the impact of a high protein diet on TMAO. Healthy men (74.2 ± 3.6 years, n = 29) were randomised to consume the recommended dietary allowance of protein (RDA: 0.8 g protein/kg bodyweight/day) or twice the RDA (2RDA) as part of a supplied diet for 10 weeks. Fasting blood samples were collected pre- and post-intervention for measurement of TMAO, blood lipids, glucose tolerance, insulin sensitivity, and inflammatory biomarkers. An oral glucose tolerance test was also performed. In comparison with RDA, the 2RDA diet increased circulatory TMAO (p = 0.002) but unexpectedly decreased renal excretion of TMAO (p = 0.003). LDL cholesterol was increased in 2RDA compared to RDA (p = 0.049), but no differences in other biomarkers of CVD risk and insulin sensitivity were evident between groups. In conclusion, circulatory TMAO is responsive to changes in dietary protein intake in older healthy males.Item Evaluation of Protein Adequacy From Plant-Based Dietary Scenarios in Simulation Studies: A Narrative Review(Elsevier Inc on behalf of the American Society for Nutrition, 2024-02) Soh BXP; Smith NW; R von Hurst P; McNabb WCAlthough a diet high in plant foods can provide beneficial nutritional outcomes, unbalanced and restrictive plant-based diets may cause nutrient deficiencies. Protein intake from these diets is widely discussed, but the comparison of animal and plant proteins often disregards amino acid composition and digestibility as measurements of protein quality. Poor provision of high-quality protein may result in adverse outcomes, especially for individuals with increased nutrient requirements. Several dietary modeling studies have examined protein adequacy when animal-sourced proteins are replaced with traditional and novel plant proteins, but no review consolidating these findings are available. This narrative review aimed to summarize the approaches of modeling studies for protein intake and protein quality when animal-sourced proteins are replaced with plant foods in diet simulations and examine how these factors vary across age groups. A total of 23 studies using dietary models to predict protein contribution from plant proteins were consolidated and categorized into the following themes-protein intake, protein quality, novel plant-based alternatives, and plant-based diets in special populations. Protein intake from plant-based diet simulations was lower than from diets with animal-sourced foods but met country-specific nutrient requirements. However, protein adequacy from some plant-sourced foods were not met for simulated diets of children and older adults. Reduced amino acid adequacy was observed with increasing intake of plant foods in some scenarios. Protein adequacy was generally dependent on the choice of substitution with legumes, nuts, and seeds providing greater protein intake and quality than cereals. Complete replacement of animal to plant-sourced foods reduced protein adequacy when compared with baseline diets and partial replacements.Item In vitro digestibility of dietary proteins and in vitro DIAAS analytical workflow based on the INFOGEST static protocol and its validation with in vivo data(Elsevier Ltd, 2023-03-15) Sousa R; Recio I; Heimo D; Dubois S; Moughan PJ; Hodgkinson SM; Portmann R; Egger LThe FAO recommends the digestible indispensable amino acid score (DIAAS) to determine protein quality in foods, preferably tested in vivo. Here, the INFOGESTin vitrodigestion protocol was applied and supplemented with an analytical workflow allowing the assessment of protein digestibility and DIAAS calculation. The protocol was applied to selected samples WPI, zein, collagen, black beans, pigeon peas, All-Bran®, and peanuts. The total protein digestibility, digestibility of individual amino acids (AA), and DIAAS values were established and compared with in vivo data for the same substrates. Total protein digestibility (total Nitrogen, r = 0.7, P < 0.05; primary amines (OPA), r = 0.6, P < 0.02; total AA, r = 0.6, P < 0.02) and digestibility of individual AA (r = 0.6, P < 0.0001) were in good agreement, between in vitro and in vivo, with a mean difference of 1.2 %. In vitro DIAAS was highly correlated with DIAAS obtained from in vivo true ileal digestibility values (r = 0.96, R2 = 0.89, P < 0.0001) with a mean difference of 0.1 %.Item Using meta-analysis to understand the impacts of dietary protein and fat content on the composition of fecal microbiota of domestic dogs (Canis lupus familiaris): A pilot study(John Wiley and Sons Ltd, 2024-04) Phimister FD; Anderson RC; Thomas DG; Farquhar MJ; Maclean P; Jauregui R; Young W; Butowski CF; Bermingham ENThe interplay between diet and fecal microbiota composition is garnering increased interest across various host species, including domestic dogs. While the influence of dietary macronutrients and their associated microbial communities have been extensively reviewed, these reviews are descriptive and do not account for differences in microbial community analysis, nor do they standardize macronutrient content across studies. To address this, a meta-analysis was performed to assess the impact of dietary crude protein ("protein") and dietary crude fat ("fat") on the fecal microbiota composition in healthy dogs. Sixteen publications met the eligibility criteria for the meta-analysis, yielding a final data set of 314 dogs. Diets were classed as low, moderate, high, or supra in terms of protein or fat content. Sequence data from each publication were retrieved from public databases and reanalyzed using consistent bioinformatic pipelines. Analysis of community diversity indices and unsupervised clustering of the data with principal coordinate analysis revealed a small effect size and complete overlap between protein and fat levels at the overall community level. Supervised clustering through random forest analysis and partial least squares-discriminant analysis indicated alterations in the fecal microbiota composition at a more individual taxonomic level, corresponding to the levels of protein or fat. The Prevotellaceae Ga6A1 group and Enterococcus were associated with increasing levels of protein, while Allobaculum and Clostridium sensu stricto 13 were associated with increasing levels of fat. Interestingly, the random forest analyses revealed that Sharpea, despite its low relative abundance in the dog's fecal microbiome, was primarily responsible for the separation of the microbiome for both protein and fat. Future research should focus on validating and understanding the functional roles of these relatively low-abundant genera.Item A randomized cross-over trial to determine the effect of a protein vs. carbohydrate preload on energy balance in ad libitum settings(BioMed Central Ltd, 2019-11-09) Gibson MJ; Dawson JA; Wijayatunga NN; Ironuma B; Chatindiara I; Ovalle F; Allison DB; Dhurandhar EJBACKGROUND: Although high protein diets have been tested in controlled environments for applications to weight management, it is not understood if adding high protein foods to the diet would impact ad libitum energy balance in the absence of other lifestyle changes. METHODS: This double-blinded randomized crossover trial compared the effects of a protein shake (PS) to a carbohydrate shake (CS), consumed prior to each major meal to equate to 20% of total energy needs over the course of the day, on energy balance over two 5-day treatment periods in healthy adults with BMI 20-30 kg/m2. Tri-axial accelerometers estimated physical activity energy expenditure. Ad libitum energy intake was measured in a laboratory kitchen. RESULTS: Energy balance was positive during both treatment periods but was not different between periods. There were no interactions between treatment and preload caloric dose or treatment and BMI status on energy balance. Satiety ratings did not differ for any pairwise comparisons between treatment and caloric dose. Controlling for gender and basal metabolic rate, thermic effect of food was greater for PS than CS. CONCLUSIONS: Preload periods significantly altered the macronutrient composition of the overall diet. This study found limited evidence that carbohydrate or protein preloads have differential effects on energy balance in short-term ad libitum settings. TRIAL REGISTRATION: This trial was pre-registered on clinicaltrials.gov as NCT02613065 on 11/30/2015.Item A higher-protein nut-based snack product suppresses glycaemia and decreases glycaemic response to co-ingested carbohydrate in an overweight prediabetic Asian Chinese cohort: the Tū Ora postprandial RCT(Cambridge University Press on behalf of The Nutrition Society, 2021-04-23) Lu LW; Silvestre MP; Sequeira IR; Plank LD; Foster M; Middleditch N; Acevedo-Fani A; Hollingsworth KG; Poppitt SDNut-based products may aid low-glycaemic dietary strategies that are important for diabetes prevention in populations at increased risk of dysglycaemia, such as Asian Chinese. This randomised cross-over trial assessed the postprandial glycaemic response (0-120 min) of a higher-protein nut-based (HP-NB) snack formulation, in bar format (1009 kJ, Nutrient Profiling Score, NPS, -2), when compared with an iso-energetic higher-carbohydrate (CHO) cereal-based bar (HC-CB, 985 kJ, NPS +3). It also assessed the ability to suppress glucose response to a typical CHO-rich food (white bread, WB), when co-ingested. Ten overweight prediabetic Chinese adults (mean, sd: age 47⋅9, 15⋅7 years; BMI 25⋅5, 1⋅6 kg/m2), with total body fat plus ectopic pancreas and liver fat quantified using dual-energy X-ray absorptiometry and magnetic resonance imaging and spectroscopy, received the five meal treatments in random order: HP-NB, HC-CB, HP-NB + WB (50 g available CHO), HC-CB + WB and WB only. Compared with HC-CB, HP-NB induced a significantly lower 30-120 min glucose response (P < 0⋅05), with an approximately 10-fold lower incremental area under the glucose curve (iAUC0-120; P < 0⋅001). HP-NB also attenuated glucose response by approximately 25 % when co-ingested with WB (P < 0⋅05). Half of the cohort had elevated pancreas and/or liver fat, with 13-21 % greater suppression of iAUC0-120 glucose in the low v. high organ fat subgroups across all five treatments. A nut-based snack product may be a healthier alternative to an energy equivalent cereal-based product with evidence of both a lower postprandial glycaemic response and modulation of CHO-induced hyperglycaemia even in high-risk, overweight, pre-diabetic adults.Item Blood Pressure and Heart Rate Responses following Dietary Protein Intake in Older Men(MDPI (Basel, Switzerland), 3/05/2022) Oberoi A; Giezenaar C; Lange K; Jones KL; Horowitz M; Chapman I; Soenen SPostprandial hypotension (PPH) occurs frequently in older people >65 years old. Protein-rich supplements, particularly whey protein (WP), are increasingly used by older people for various health benefits. We have reported that 70 g WP drinks cause significant, and in some cases marked, falls in blood pressure (BP) in older men. The effects of lower, more widely used, doses (~30 g) on systolic (SBP) and diastolic (DBP) blood pressure and heart rate (HR) are not known. In a randomized order, eight older men (age: 72 ± 1 years; body mass index (BMI): 25 ± 1 kg/m2) after overnight fast ingested a drink containing (i) a non-caloric control (~2 kcal), (ii) 30 g of whey protein (120 kcal; ‘WP30’), or (iii) 70 g of whey protein (280 kcal; ‘WP70’). The BP and HR were measured in this pilot study with an automated device before and at 3-min intervals for 180 min following drink ingestion. Drink condition effects were determined by repeated-measures ANOVA. The SBP decreased after both WP drinks compared to the control (p = 0.016), particularly between 120 and 180 min, with no difference in the effects of WP30 and WP70. The SBP decreased by ≥20 mmHg in more than 50% of people after both WP drinks (WP30: 63%; WP70: 75%) compared to 38% after the control. The maximum fall in the SBP occurred during the third hour, with the nadir occurring latest after WP70. The DBP decreased non-significantly by several mmHg more after the WP drinks than after the control. The maximum HR increases occurred during the third hour, with the greatest increase after WP70. The SBP decreased after both WP drinks compared to the control, with the effects most evident between 120 and 180 min. Accordingly, ingestion of even relatively modest protein loads in older men has the potential to cause PPH.