Alastair B. Ross

Systemic multicompartmental effects of the gut microbiome on mouse metabolic phenotypes

To characterize the impact of gut microbiota on host metabolism, we investigated the multicompartmental metabolic profiles of a conventional mouse strain (C3H/HeJ) (n=5) and its germ‐free (GF) equivalent (n=5). We confirm that the microbiome strongly impacts on the metabolism of bile acids through the enterohepatic cycle and gut metabolism (higher levels of phosphocholine and glycine in GF liver and marked higher levels of bile acids in three gut compartments). Furthermore we demonstrate that (1) well‐defined metabolic differences exist in all examined compartments between the metabotypes of GF and conventional mice: bacterial co‐metabolic products such as hippurate (urine) and 5‐aminovalerate (colon epithelium) were found at reduced concentrations, whereas raffinose was only detected in GF colonic profiles. (2) The microbiome also influences kidney homeostasis with elevated levels of key cell volume regulators (betaine, choline, myo‐inositol and so on) observed in GF kidneys. (3) Gut microbiota modulate metabotype expression at both local (gut) and global (biofluids, kidney, liver) system levels and hence influence the responses to a variety of dietary modulation and drug exposures relevant to personalized health‐care investigations.

Colonization-Induced Host-Gut Microbial Metabolic Interaction

ABSTRACT The gut microbiota enhances the host’s metabolic capacity for processing nutrients and drugs and modulate the activities of multiple pathways in a variety of organ systems. We have probed the systemic metabolic adaptation to gut colonization for 20 days following exposure of axenic mice (n = 35) to a typical environmental microbial background using high-resolution 1H nuclear magnetic resonance (NMR) spectroscopy to analyze urine, plasma, liver, kidney, and colon (5 time points) metabolic profiles. Acquisition of the gut microbiota was associated with rapid increase in body weight (4%) over the first 5 days of colonization with parallel changes in multiple pathways in all compartments analyzed. The colonization process stimulated glycogenesis in the liver prior to triggering increases in hepatic triglyceride synthesis. These changes were associated with modifications of hepatic Cyp8b1 expression and the subsequent alteration of bile acid metabolites, including taurocholate and tauromuricholate, which are essential regulators of lipid absorption. Expression and activity of major drug-metabolizing enzymes (Cyp3a11 and Cyp2c29) were also significantly stimulated. Remarkably, statistical modeling of the interactions between hepatic metabolic profiles and microbial composition analyzed by 16S rRNA gene pyrosequencing revealed strong associations of the Coriobacteriaceae family with both the hepatic triglyceride, glucose, and glycogen levels and the metabolism of xenobiotics. These data demonstrate the importance of microbial activity in metabolic phenotype development, indicating that microbiota manipulation is a useful tool for beneficially modulating xenobiotic metabolism and pharmacokinetics in personalized health care. IMPORTANCE Gut bacteria have been associated with various essential biological functions in humans such as energy harvest and regulation of blood pressure. Furthermore, gut microbial colonization occurs after birth in parallel with other critical processes such as immune and cognitive development. Thus, it is essential to understand the bidirectional interaction between the host metabolism and its symbionts. Here, we describe the first evidence of an in vivo association between a family of bacteria and hepatic lipid metabolism. These results provide new insights into the fundamental mechanisms that regulate host-gut microbiota interactions and are thus of wide interest to microbiological, nutrition, metabolic, systems biology, and pharmaceutical research communities. This work will also contribute to developing novel strategies in the alteration of host-gut microbiota relationships which can in turn beneficially modulate the host metabolism. Gut bacteria have been associated with various essential biological functions in humans such as energy harvest and regulation of blood pressure. Furthermore, gut microbial colonization occurs after birth in parallel with other critical processes such as immune and cognitive development. Thus, it is essential to understand the bidirectional interaction between the host metabolism and its symbionts. Here, we describe the first evidence of an in vivo association between a family of bacteria and hepatic lipid metabolism. These results provide new insights into the fundamental mechanisms that regulate host-gut microbiota interactions and are thus of wide interest to microbiological, nutrition, metabolic, systems biology, and pharmaceutical research communities. This work will also contribute to developing novel strategies in the alteration of host-gut microbiota relationships which can in turn beneficially modulate the host metabolism.

Whole Grain Compared with Refined Wheat Decreases the Percentage of Body Fat Following a 12-Week, Energy-Restricted Dietary Intervention in Postmenopausal Women

Observational studies show inverse associations between intake of whole grain and adiposity and cardiovascular risk; however, only a few dietary intervention trials have investigated the effect of whole-grain consumption on health outcomes. We studied the effect of replacing refined wheat (RW) with whole-grain wheat (WW) for 12 wk on body weight and composition after a 2-wk run-in period of consumption of RW-containing food intake. In this open-label randomized trial, 79 overweight or obese postmenopausal women were randomized to an energy-restricted diet (deficit of ~1250 kJ/d) with RW or WW foods providing 2 MJ/d. Body weight and composition, blood pressure, and concentration of circulating risk markers were measured at wk 0, 6, and 12. Fecal output and energy excretion were assessed during run-in and wk 12. Plasma alkylresorcinol analysis indicated good compliance with the intervention diets. Body weight decreased significantly from baseline in both the RW (-2.7 ± 1.9 kg) and WW (-3.6 ± 3.2 kg) groups, but the decreases did not differ between the groups (P = 0.11). The reduction in body fat percentage was greater in the WW group (-3.0%) than in the RW group (-2.1%) (P = 0.04). Serum total and LDL cholesterol increased by ~5% (P < 0.01) in the RW group but did not change in the WW group; hence, the changes differed between the groups (P = 0.02). In conclusion, consumption of whole-grain products resulted in a greater reduction in the percentage fat mass, whereas body weight changes did not differ between the RW and WW groups. Serum total and LDL cholesterol, two important risk factors of cardiovascular disease, increased with RW but not WW consumption, which may suggest a cardioprotective role for whole grain.

Whole grain and body weight changes in apparently healthy adults: a systematic review and meta-analysis of randomized controlled studies

BACKGROUND Whole grains are recognized for their potential role in preventing cardiovascular diseases; however, results from randomized controlled studies on blood lipids are inconsistent, potentially because of compositional differences between individual grain types for some nutrients, including dietary fiber. OBJECTIVE Using a meta-analytic approach, we assessed the effect of whole-grain compared with non-whole-grain foods on changes in total cholesterol (TC), LDL cholesterol, HDL cholesterol, and triglycerides. DESIGN We conducted a systematic literature search in selected databases. Studies were included if they were randomized controlled comparisons between whole-grain foods and a non-whole-grain control in adults. A total of 6069 articles were screened for eligibility, and data were extracted from 24 studies. Weighted mean differences were calculated, and meta-regression analyses were performed for whole-grain dose, study duration, and baseline TC concentration. RESULTS Overall, whole-grain intake lowered LDL cholesterol (weighted difference: -0.09 mmol/L; 95% CI: -0.15, -0.03 mmol/L; P < 0.01) and TC (weighted difference: -0.12 mmol/L; 95% CI: -0.19, -0.05 mmol/L; P < 0.001) compared with the control. Whole-grain oat had the greatest effect on TC (weighted difference: -0.17 mmol/L; 95% CI: -0.10, -0.25 mmol/L; P < 0.001). No effect of whole-grain foods on HDL cholesterol was seen, whereas whole-grain foods tended to lower triglycerides compared with the control (weighted difference: -0.04 mmol/L; 95% CI: -0.08, 0.01; P = 0.10). No association was found between whole-grain dose or baseline TC concentration and any of the outcomes, whereas study duration was positively associated with changes in TC and LDL cholesterol. CONCLUSIONS Consumption of whole-grain diets lowers LDL cholesterol and TC, but not HDL cholesterol or triglycerides, compared with consumption of non-whole-grain control diets. Whole-grain oat appears to be the most effective whole grain for lowering cholesterol.

A whole-grain cereal-rich diet increases plasma betaine, and tends to decrease total and LDL-cholesterol compared with a refined-grain diet in healthy subjects

Epidemiological studies have repeatedly found that whole-grain (WG) cereal foods reduce the risk of several lifestyle-related diseases, though consistent clinical outcomes and mechanisms are elusive. To compare the effects of a WG-rich diet with a matched refined-grain (RG) diet on plasma biomarkers and bowel health parameters, seventeen healthy subjects (eleven females and six males) completed an exploratory cross-over study with a 2-week intervention diet based on either WG- or RG-based foods, separated by a washout of at least 5 weeks. Both diets were the same except for the use of WG (150 g/d) or RG foods. Subjects undertook a 4 h postprandial challenge on day 8 of each intervention diet. After 2 weeks, the WG diet tended to decrease plasma total and LDL-cholesterol (both P = 0·09), but did not change plasma HDL-cholesterol, fasting glucose, C-reactive protein or homocysteine compared with the RG diet. Plasma betaine and alkylresorcinol concentrations were elevated after 1 week of the WG diet (P = 0·01 and P < 0·0001, respectively). Clostridium leptum populations in faeces were increased after the WG diet, along with a trend for decreased faecal water pH (P = 0·096) and increased stool frequency (P < 0·0001) compared with the RG diet. A short controlled intervention trial with a variety of commercially available WG-based products tended to improve biomarkers of CVD compared with a RG diet. Changes in faecal microbiota related to increased fibre fermentation and increased plasma betaine concentrations point to both fibre and phytochemical components of WG being important in mediating any potential health effects.

Present Status and Perspectives on the Use of Alkylresorcinols as Biomarkers of Wholegrain Wheat and Rye Intake

Alkylresorcinols (ARs) were first proposed as potential biomarkers of wholegrain wheat and rye intake a decade ago. Since then there has been a considerable body of research which suggests that ARs do meet most criteria of a biomarker of these foods. Results from human studies on plasma AR and their plasma and urinary metabolites strongly indicate that these compounds are responsive to whole grain wheat and rye intake and are correlated with various measures of AR consumption. This review briefly summarises work on the bioactivities of AR and focuses on aspects related to their use as biomarkers of whole grain wheat and rye intake. Evidence suggests that they thus far broadly fulfil the criteria to act as biomarkers of these cereals. However, there are still gaps in the knowledge on factors relating to the wide interindividual variation, and application to different epidemiological cohorts. Overall, ARs are highly promising biomarkers of whole grain wheat and rye intake and add to our increasing understanding of whole grains and health.

Plasma alkylresorcinols as a biomarker of whole-grain food consumption in a large population: results from the WHOLEheart Intervention Study

BACKGROUND Small-scale, short-term intervention studies have suggested that plasma alkylresorcinol (AR) concentrations may be biomarkers of whole grain (WG) wheat and rye intakes. OBJECTIVE The objective was to determine whether plasma AR concentrations reflect self-reported WG food intake in a 16-wk WG intervention study and to establish which phenotypic characteristics influence plasma AR concentrations. DESIGN In a randomized parallel-group dietary intervention study, 316 overweight and obese participants with a WG intake of <30 g/d were recruited and randomly assigned to 1 of 3 groups: control (no dietary change), intervention 1 (60 g WG/d for 16 wk), or intervention 2 (60 g WG/d for 8 wk followed by 120 g WG/d for 8 wk). Fasting blood samples were collected at baseline, 8 wk, and 16 wk for the measurement of plasma lipids and ARs. RESULTS Plasma samples from 266 study completers were analyzed. Total plasma AR concentrations increased with the WG intervention and could be used to distinguish between control subjects and those who consumed 60 or 120 g WG, but not between those who consumed 60 and 120 g WG. Plasma AR concentrations were higher in men, were positively associated with plasma triglyceride concentrations, and were negatively associated with nonesterified fatty acids. CONCLUSIONS Plasma AR concentrations were correlated with WG intake and could be used to distinguish between low- and high-WG consumers. Sex and plasma lipid concentrations independently influenced plasma AR concentrations, although plasma triglycerides may explain higher concentrations in men. This trial is registered as ISRCT no. 83078872.

Rapid and Sensitive Analysis of Alkylresorcinols from Cereal Grains and Products Using HPLC―Coularray-Based Electrochemical Detection

Alkylresorcinols (AR) present in the bran fraction of wheat, rye, and barley grains are of current interest as biomarkers of wholegrain cereal intake. This paper reports the development of a rapid quantitative HPLC method allowing sensitive analysis of AR in cereals in 14 min using coularray (CA) detection. The LOD was 1 ng injected and the LOQ 2.5 ng injected, whereas the intrarun and inter-run CVs were 0.8 and 6.4%, respectively. Analytical recovery determined by spiking ranged from 98 to 107% for AR homologues C15:0-C25:0. The method was used to analyze 79 flour and cereal products, with results in the same range as previous studies using GC analysis. Analysis of wholegrain white wheat were in the same range as other wheat varieties (420-556 microg/g). Use of the CA detector allowed detection of low concentrations of AR in white wheat flour (13-51 microg/g) that had not been reliably detected using previous methods. This method allows rapid throughput analysis of cereal samples, required for validation studies on the use of AR as biomarkers of wholegrain cereal intake.

Quantitative Measurement of Betaine and Free Choline in Plasma, Cereals and Cereal Products by Isotope Dilution LC-MS/MS

Betaine and choline are important components of the one-carbon metabolism cycle, linked with the amino acid homocysteine and lipid metabolism. Analyses of broad ranges of foods point to cereal based foods being important sources of betaine and choline, however to date there has been no detailed analysis of these compounds in cereal flours or cereal products. An analytical method based on optimization of an existing extraction followed by LC-MS/MS analysis was used to analyze 47 plasma samples, 32 cereal flours and cereal fractions, and 51 cereal products. For the method validation LLOQ, recovery, inter- and intraday repeatability were all performed. Whole-grain wheat and rye flours, and products based on these were the best whole cereal sources of betaine (747-1508 microg/g) and to a lesser extent choline (76-159 microg/g), while the bran fraction contained the highest concentrations of betaine and free-choline (2350-2899 microg/g and 366-384 microg/g respectively). Refined wheat flour and products contained lower concentrations, while rice and maize contained only very low and no detectable amounts of betaine respectively (0-10 microg/g), and low amounts of free-choline (<31 microg/g). These results were mirrored in cereal products analyzed, with whole-grain wheat or rye-based cereal products having the highest concentrations of the two metabolites. Plasma concentrations for betaine and free-choline in a group of 47 subjects ranged from 15.2-66.3 and 9.8-18.5 micromol/L respectively, within the range of previous reports. This LC-MS/MS method can be used to rapidly and sensitively quantify betaine and free-choline in plasma and cereal products. Whole-grain cereal products and products containing cereal bran appear to be excellent dietary sources of betaine and free-choline.

Intake of alkylresorcinols from wheat and rye in the United Kingdom and Sweden

Alkylresorcinols, phenolic lipids present in high amounts in whole-grain wheat and rye but not present in appreciable amounts in other foods, are candidates as biomarkers of whole-grain intake from these cereals. We estimated the intake of alkylresorcinols in Sweden and the UK using two different methods: food supply data (FSD); food consumption data (FCD; based on individual and household survey data). The average per capita intake of alkylresorcinols in Sweden was 17.5 mg/d (FSD) and 22.9 (sd 16.6) mg/d (FCD), while in the UK it was 11.9 mg/d (FSD) and 11.8 (sd 18.62) mg/d (FCD). Ninety-six per cent of all Swedes consumed some alkylresorcinols, compared with 50 % of British people surveyed. Both women and men over the age of 40 years had a higher alkylresorcinol intake than younger people. The average results from the two methods were similar, but the FCD data provided more detail about the range of alkylresorcinol intake, and indicate that because the intake of alkylresorcinols varies so widely, they may be good markers of diets rich or poor in whole-grain wheat/rye products.