Fecal Metagenomics to Identify Biomarkers of Food Intake in Healthy Adults: Findings from Randomized, Controlled, Nutrition Trials.

BACKGROUND: Undigested components of the human diet affect the composition and function of the microorganisms present in the gastrointestinal tract. Techniques like metagenomic analyses allow researchers to study functional capacity, thus revealing the potential of using metagenomic data for developing objective biomarkers of food intake. OBJECTIVES: As a continuation of our previous work using 16S and metabolomic datasets, we aimed to utilize a computationally intensive, multivariate, machine-learning approach to identify fecal KEGG (Kyoto encyclopedia of genes and genomes) Orthology (KO) categories as biomarkers that accurately classify food intake. METHODS: Data were aggregated from 5 controlled feeding studies that studied the individual impact of almonds, avocados, broccoli, walnuts, barley, and oats on the adult gastrointestinal microbiota. Deoxyribonucleic acid from preintervention and postintervention fecal samples underwent shotgun genomic sequencing. After preprocessing, sequences were aligned and functionally annotated with Double Index AlignMent Of Next-generation sequencing Data v2.0.11.149 and MEtaGenome ANalyzer v6.12.2, respectively. After the count normalization, the log of the fold change ratio for resulting KOs between pre- and postintervention of the treatment group against its corresponding control was utilized to conduct differential abundance analysis. Differentially abundant KOs were used to train machine-learning models examining potential biomarkers in both single-food and multi-food models. RESULTS: We identified differentially abundant KOs in the almond (n = 54), broccoli (n = 2474), and walnut (n = 732) groups (q < 0.20), which demonstrated classification accuracies of 80%, 87%, and 86% for the almond, broccoli, and walnut groups using a random forest model to classify food intake into each food group's respective treatment and control arms, respectively. The mixed-food random forest achieved 81% accuracy. CONCLUSIONS: Our findings reveal promise in utilizing fecal metagenomics to objectively complement self-reported measures of food intake. Future research on various foods and dietary patterns will expand these exploratory analyses for eventual use in feeding study compliance and clinical settings.

Honey Added to Yogurt with Bifidobacterium animalis subsp. lactis DN-173 010/CNCM I-2494 Supports Probiotic Enrichment but Does Not Reduce Intestinal Transit Time in Healthy Adults: A Randomized, Controlled, Crossover Trial.

BACKGROUND: Honey improves probiotic survival in vitro. However, if this effect translates to humans has not been investigated. OBJECTIVES: We aimed to determine effects of honey plus yogurt containing the probiotic Bifidobacterium animalis subsp. lactis DN-173 010/CNCM I-2494 (B. animalis) on intestinal transit time, probiotic enrichment, digestive health, mood, and cognition in adults. METHODS: Sixty-six healthy adults (34 female; 33.6 ± 9.8 y; 24.6 ± 3.0 kg/m2) in a crossover trial were randomly assigned to 2-wk yogurt conditions in a counterbalanced order with ≥4-wk washout: 1) Honey (HON): yogurt plus honey and 2) Negative Control (NC): heat-treated yogurt plus sugar. Of the participants, n = 62 completed the trial, and n = 37 (17 female; 32.0 ± 8.3 y; 25.0 ± 2.9 kg/m2) elected to enroll in a third condition (a nonrandomized study extension) after ≥4-wk washout with a reference Positive Control (PC): yogurt plus sugar. At baseline and end of each of the 3 conditions, intestinal transit time was measured with dye capsules; probiotic abundance with fecal DNA 16S sequencing; digestive health with symptom/function records, Bristol stool consistency, Gastrointestinal Tolerability, and Gastrointestinal Quality of Life Index; mood with Positive and Negative Affect Schedule-Short Form, Depression Anxiety Stress Scales-42, Patient-Reported Outcomes Measurement Information System questionnaires, and an emotional image task; and cognition with a spatial reconstruction task. Data were analyzed using linear mixed-effects models (LMMs) with significance at P ≤ 0.05. Baseline and end data were included in the LMM, with fixed effects being treatment, time, treatment by time interaction, and baseline covariate, and the random effect being the participant. RESULTS: B. animalis was enriched in HON (d = 3.54; P = 0.0002) compared to controls with linear discriminant analysis effect size. Intestinal transit time, gastrointestinal health, mood, and cognition did not differ between conditions (LMM: Ps > 0.05). CONCLUSIONS: Yogurt + honey enriched B. animalis but did not reduce intestinal transit time or have other functional gastrointestinal, mood, or cognitive effects in adults. This trial was registered at www. CLINICALTRIALS: gov as NCT04187950 and NCT04901390.

Skin and macular carotenoids and relations to academic achievement among school-aged children.

ABSTRACTObjectives: Carotenoids are plant pigments that accumulate in human tissue (e.g. macula and skin) and can serve as biomarkers for diet quality; however, knowledge on skin and macular carotenoids in relation to cognition in children is limited. This study aimed to address this gap by assessing links between skin and macular carotenoids and academic achievement in school-aged children.Methods: Children 7-12 years old (n = 81) participated in a crosssectional study. Skin and macular carotenoids were measured with reflection spectroscopy and heterochromatic flicker photometry, respectively. Academic achievement was measured using Woodcock-Johnson IV (WJ-IV). Body Mass Index was calculated using height and weight measurements, demographic information was collected using a family demographics and pediatric health history questionnaire, and carotenoid intake was assessed using 7-day diet records.Results: Skin carotenoids were not related to macular pigment (r = 0.08, p = 0.22). Adjusting for age, sex, BMI percentile, household income, and total carotenoid consumption (mg/1000kcal), skin carotenoids were predictive of math (ß = 0.27, p = 0.02), broad math (ß = 0.36, p < 0.01) and math calculation (ß = 0.38, p < 0.01). Skin carotenoids displayed trending relationships with broad reading (ß = 0.23, p = 0.05) and reading fluency (ß = 0.22, p = 0.07). There were no significant associations between macular pigment and academic achievement (all ß's ≤ 0.07, all p's ≥ 0.56).Discussion: Skin carotenoids were positively associated with academic abilities in children, while macular carotenoids did not display this relationship. Future interventions examining prospective effects of changes in carotenoids in different tissues on childhood academic achievement are warranted.

Dietary fiber intake and fecal short-chain fatty acid concentrations are associated with lower plasma lipopolysaccharide-binding protein and inflammation.

Consuming adequate dietary fiber is a promising strategy for reducing systemic inflammation. The objective was to evaluate relationships between dietary fiber intake, markers of metabolic endotoxemia, and systemic inflammation in adults. This was a cross-sectional study of 129 healthy participants (age 33.6 ± 6.1 yr, BMI 30.5 ± 6.9 kg/m2). Dietary fiber intake was assessed by food frequency questionnaire. Adiposity was measured using dual-energy X-ray absorptiometry (DXA). Fecal short-chain fatty acids (SCFA) were quantified using gas chromatography-mass spectrometry. Fecal microbiota sequence data (V4 region, 16S rRNA gene) were analyzed using DADA2 and QIIME2. Inflammatory cytokines were assessed with enzyme-linked immunosorbent assays; flow cytometry was conducted for monocyte surface marker quantification. Bivariate correlations and generalized step-wise linear modeling were used for statistical analyses. Plasma C-reactive protein (CRP) and interleukin (IL)-6 concentrations were positively related to whole body (CRP r = 0.45, P = <0.0001; IL-6 r = 0.34, P = 0.0002) and visceral adiposity (CRP r = 0.33, P = 0.0003; IL-6 r = 0.38, P = 0.0002). Plasma lipopolysaccharide-binding protein (LBP) concentrations were inversely related to dietary fiber intake (r = -0.22, P = 0.03) and fecal SCFA (acetate r = -0.25, P = 0.01; propionate r = -0.28, P = 0.003; butyrate r = -0.23, P = 0.02). Whole body adiposity, dietary fiber, and fecal SCFA were the most predictive of plasma LBS-BP concentrations. Novel findings included associations between dietary fiber intake, the gastrointestinal microbiota, and systemic inflammation.NEW & NOTEWORTHY Dietary fiber intake may reduce the inflammation associated with obesity and metabolic disease. Our cross-sectional analysis revealed that dietary fiber intake and fecal short-chain fatty acids are inversely associated with lipopolysaccharide-binding protein, a marker of systemic inflammation. In addition, plasma interleukin-6 and C-reactive protein were positively related to markers of adiposity.

Adherence to Screen Time and Physical Activity Guidelines is Associated with Executive Function in US Toddlers Participating in the STRONG Kids 2 Birth Cohort Study.

OBJECTIVE: To test the hypothesis that healthy weight status and adherence to American Academy of Pediatrics (AAP) guidelines for diet and physical activity would extend to greater executive function (EF) at age 24 months. STUDY DESIGN: Parents of 24-month-old children from the STRONG Kids 2 cohort study (n = 352) completed the Behavioral Rating Inventory of Executive Function for Preschoolers (BRIEF-P) and reported physical activities, diet, and screen time. Toddlers met AAP guidelines if they consumed at least 5 servings of fruits and vegetables, were physically active, refrained from sugar-sweetened beverages, and limited daily screen time to <60 minutes. Relationships between EF, 24-month weight status, and meeting AAP guidelines were tested independent of child sex, ethnicity, socioeconomic status, weight status at birth, and maternal pregnancy weight status. RESULTS: Weight-for-length z-score had no effect on EF. Toddlers meeting the screen time guideline had greater EF (ß, -0.125; 95% CI, 0.234 to -0.008), inhibitory self-control (ß, -0.142; 95% CI, -0.248 to -0.029), and emergent metacognition (ß, -0.111; 95% CI, -0.221 to 0.002), indicated by lower BRIEF-P scores. Those with more minutes of screen time had poorer overall EF (ß, 0.257; 95% CI, 0.118-0.384), inhibitory self-control (ß, 0.231; 95% CI, 0.099-0.354), cognitive flexibility (ß, 0.217; 95% CI, 0.082-0.342), and emergent metacognition (ß, 0.257; 95% CI, 0.120-0.381). Daily physical activity was associated with greater emergent metacognition (ß, -0.116; 95% CI, -0.225 to -0.005). CONCLUSIONS: Meeting AAP guidelines for physical activity and screen time was related to greater EF in a demographically homogenous sample of toddlers. Future randomized control trials and more diverse samples are needed to confirm the directionality of this relationship. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov, NCT03341858.

Prebiotic Consumption Alters Microbiota but Not Biological Markers of Stress and Inflammation or Mental Health Symptoms in Healthy Adults: A Randomized, Controlled, Crossover Trial.

BACKGROUND: Chronic stress contributes to systemic inflammation and diminished mental health. Although animal work suggests strong links with the microbiota-gut-brain axis, clinical trials investigating the effectiveness of prebiotics in improving mental health and reducing inflammation are lacking. OBJECTIVES: We aimed to determine fructooligosaccharide (FOS) and galactooligosaccharide (GOS) effects on biological markers of stress and inflammation and mental health symptoms in adults. Secondary outcomes included fecal microbiota and metabolites, digestive function, emotion, and sleep. METHODS: Twenty-four healthy adults (25-45 y; 14 females, 10 males; BMI, 29.3 ± 1.8 kg/m2) from central Illinois participated in a 2-period, randomized, controlled, single-blinded crossover trial. Interventions included the prebiotic (PRE) treatment (237 mL/d Lactaid low-fat 1% milk, 5 g/d FOS, 5 g/d GOS) and control (CON) (237 mL/d Lactaid), which were consumed in counterbalanced order for 4 wk each, separated by ≥4-wk washout. Inflammatory markers were measured in blood plasma (>10-h fast) and cortisol in urine. The Depression Anxiety Stress Scales-42 assessed mental health symptoms. Fecal samples were collected for 16S rRNA gene (V4 region) sequencing and analysis. Emotion was measured by rating images from a computer task. Sleep was assessed using 7-d records and accelerometers. Change scores were analyzed using linear mixed models with treatment and baseline covariate as fixed effects and participant ID as the random effect. RESULTS: There were no differences in change scores between PRE and CON treatments on biological markers of stress and inflammation or mental health. PRE increased change in percent sequences (q = 0.01) of Actinobacteriota (CON: 0.46 ± 0.70%; PRE: 5.40 ± 1.67%) and Bifidobacterium (CON: -1.72 ± 0.43%; PRE: 4.92 ± 1.53%). There were also no differences in change scores between treatments for microbial metabolites, digestive function, emotion, or sleep quality. CONCLUSIONS: FOS+GOS did not affect biological markers of stress and inflammation or mental health symptoms in healthy adults; however, it increased Bifidobacterium. CLINICAL TRIAL REGISTRY: NCT04551937, www. CLINICALTRIALS: gov.

Fecal Metabolites as Biomarkers for Predicting Food Intake by Healthy Adults.

BACKGROUND: The fecal metabolome is affected by diet and includes metabolites generated by human and microbial metabolism. Advances in -omics technologies and analytic approaches have allowed researchers to identify metabolites and better utilize large data sets to generate usable information. One promising aspect of these advancements is the ability to determine objective biomarkers of food intake. OBJECTIVES: We aimed to utilize a multivariate, machine learning approach to identify metabolite biomarkers that accurately predict food intake. METHODS: Data were aggregated from 5 controlled feeding studies in adults that tested the impact of specific foods (almonds, avocados, broccoli, walnuts, barley, and oats) on the gastrointestinal microbiota. Fecal samples underwent GC-MS metabolomic analysis; 344 metabolites were detected in preintervention samples, whereas 307 metabolites were detected postintervention. After removing metabolites that were only detected in either pre- or postintervention and those undetectable in ≥80% of samples in all study groups, changes in 96 metabolites relative concentrations (treatment postintervention minus preintervention) were utilized in random forest models to 1) examine the relation between food consumption and fecal metabolome changes and 2) rank the fecal metabolites by their predictive power (i.e., feature importance score). RESULTS: Using the change in relative concentration of 96 fecal metabolites, 6 single-food random forest models for almond, avocado, broccoli, walnuts, whole-grain barley, and whole-grain oats revealed prediction accuracies between 47% and 89%. When comparing foods with one another, almond intake was differentiated from walnut intake with 91% classification accuracy. CONCLUSIONS: Our findings reveal promise in utilizing fecal metabolites as objective complements to certain self-reported food intake estimates. Future research on other foods at different doses and dietary patterns is needed to identify biomarkers that can be applied in feeding study compliance and clinical settings.

Randomized, Placebo-Controlled, Single-Blind Study of Lutein Supplementation on Carotenoid Status and Cognition in Persons with Multiple Sclerosis.

BACKGROUND: Multiple sclerosis (MS) is traditionally managed using disease-modifying pharmaceutical therapies as a first line approach for treatment, yet there is increasing interest in lifestyle factors, particularly diet, for managing disease outcomes. Lutein has neuroprotective properties in healthy adults, but no previous research has examined the effects of lutein supplementation in persons with MS. OBJECTIVES: This study aimed to investigate the efficacy of 4-mo lutein supplementation on carotenoid status and cognition in persons with relapse-remitting MS (RRMS). METHODS: A randomized controlled, single-blind research design was used among adults with RRMS (N = 21). Participants were randomized into placebo (n = 9) or treatment (20-mg/d lutein, n = 12) groups with outcomes measured before and after 4 mo. Macular pigment optical density (MPOD) was assessed using heterochromatic flicker photometry. Skin carotenoids were assessed using reflection spectroscopy. Serum lutein was measured using high-performance liquid chromatography. Cognition was assessed via the Eriksen flanker with event-related potentials, spatial reconstruction, and the symbol digit modalities tests. RESULTS: There was a significant group by time interaction for MPOD (F = 6.74, P = 0.02), skin carotenoids (F = 17.30, P < 0.01), and serum lutein (F = 24.10, P < 0.01), whereby the treatment group improved in all carotenoid outcomes. There were no significant group by time interactions for cognitive and neuroelectric outcomes. However, increase in MPOD was positively associated with accuracy during the flanker incongruent trials (r = 0.55, P = 0.03) and the spatial memory task (r = 0.58, P = 0.02) among treatment participants. CONCLUSIONS: Lutein supplementation increases carotenoid status among persons with RRMS. There is no significant effect on cognitive function but change in macular carotenoids is selectively associated with improved attention and memory. This study provides preliminary support for a fully powered study targeting retinal and neural carotenoids for cognitive benefits in persons with MS. This trial was registered at clinicaltrials.gov as NCT04843813.

MIND Dietary Pattern Adherence Is Selectively Associated with Cognitive Processing Speed in Middle-Aged Adults.

BACKGROUND: Most previous work has focused on a single-nutrient, bottom-up approach when examining the influence of diet on neurocognition. Thus, the impact of dietary patterns on cognitive health is underinvestigated. OBJECTIVES: We aimed to investigate the relation between different diet indices [i.e., Mediterranean, Dietary Approaches to Stop Hypertension (DASH), Healthy Eating Index-2015 (HEI-2015), and Mediterranean-DASH Intervention for Neurodegenerative Delay (MIND)] and attentional inhibition and neuroelectric function. METHODS: We undertook a cross-sectional analysis of adults aged 34.1 ± 6.0 y (n = 207, 116 females) with a BMI of 18.5 to >40 kg/m2 who completed the Dietary History Questionnaire II (DHQII) FFQ (Past Month and Year with Serving Sizes) to assess adherence to different diet quality indices. Attentional inhibition was assessed using a modified Eriksen flanker task during which event-related potentials (ERPs) were recorded. The amplitude and latency of the P3/P300 ERP were used to index attentional resource allocation and information processing speed, respectively. RESULTS: P3 peak latency was inversely associated with greater adherence to the MIND dietary pattern during incongruent flanker trials (∆R2 = 0.02, ß = -0.14, P = 0.043) but not during congruent trials. Adherence to the Mediterranean, DASH, and HEI-2015 patterns was not associated with P3 latency (P > 0.05). No associations were observed between the diet indices and attentional inhibition at the behavioral level (i.e., accuracy or reaction time) or P3 amplitude (all P values >0.05). CONCLUSIONS: Greater adherence to the MIND diet was selectively associated with faster information processing speed in middle-aged adults with healthy to obese BMI. Further, the influence of the MIND diet for faster information processing speed might be particularly beneficial when cognitive control demands are increased. Future intervention trials testing the effects of consuming a MIND diet on cognitive function are warranted to help inform dietary recommendations for healthy cognitive aging.

Macular Pigment Optical Density and Skin Carotenoids in a Childhood Sample.

BACKGROUND: Carotenoids are plant pigments with light filtering and antioxidant properties that deposit in human tissues, including retina and skin. Descriptive characteristics and covariates of carotenoid status in macula and skin have been examined in adults; however, similar studies in children are limited. Thus, this study aimed to delineate how factors of age, sex, race, weight status, and dietary carotenoid intake relate to macular and skin carotenoids in children. METHODS: Children (7-13 y, N = 375) completed heterochromatic flicker photometry to assess macular pigment optical density (MPOD). Participants underwent anthropometrics to measure weight status (BMI percentile [BMI%]), and parent/guardian provided demographic information. Subsample data were available for skin carotenoids (N = 181), assessed using reflection spectroscopy, and dietary carotenoids (N = 101) using the Block Food Frequency Questionnaire. Relationships between skin and macular carotenoids were assessed using partial Pearson's correlations controlling for age, sex, race, and BMI%. Relationships between dietary carotenoids and macular and skin carotenoids were assessed using stepwise linear regression including age, sex, race, and BMI% in the model. RESULTS: Mean MPOD was 0.56 ± 0.22 and skin carotenoid score was 282 ± 94.6. There was no significant correlation between MPOD and skin carotenoids (r = 0.02, P = 0.76). BMI% was negatively associated with skin (stdß = -0.42, P < 0.001), but not macular carotenoids (stdß = -0.04, P = 0.70). Neither MPOD nor skin carotenoids were associated with age, sex, or race (all P > 0.10). MPOD was positively associated with energy-adjusted reported lutein + zeaxanthin intake (stdß = 0.27, P = 0.01). Skin carotenoids were positively associated with energy-adjusted reported carotenoid intake (stdß = 0.26, P = 0.01). CONCLUSIONS: The mean MPOD values in children were higher than what has been reported in adult populations. Previous studies in adult samples report an average MPOD of 0.21. Although macular and skin carotenoids were not related, they were associated with dietary carotenoids relevant to the respective tissues; however, skin carotenoids may be more susceptible negative influence from higher weight status.

Consumption of a fermented dairy beverage improves hippocampal-dependent relational memory in a randomized, controlled cross-over trial.

Objectives: We aimed to determine whether consumption of a fermented dairy beverage containing probiotic microorganisms influences negative mood states, stress, and hippocampal memory performance in healthy adults. Methods: Adults (25-45 yrs, N = 26) free of gastrointestinal and mental illness were enrolled in a single-blind, randomized, controlled, crossover trial. Participants completed testing prior to and after 4-week consumption, with a 2-4 week washout between treatments of: (1) 8 oz of a dairy-based fermented beverage containing 25-30 billion colony forming units of live and active kefir cultures or (2) 8 oz isocaloric, non-fermented, 1% low-fat lactose-free dairy-based control beverage. Hippocampal-dependent relational memory was assessed using a spatial reconstruction task. Negative mood states of depression and anxiety were assessed using the Depression Anxiety Stress Scales-42 (DASS-42). Pooled 24-hour urine samples were analyzed using an enzyme-linked immunosorbent assay to determine urinary free-cortisol (UFC) concentrations. Fecal microbiota composition was assessed using 16S rRNA gene sequencing. Results: Lactobacillus was increased by 235% following fermented dairy consumption compared to the control (p < .01). Furthermore, the fermented dairy beverage improved performance on two metrics of relational memory, misplacement (p = .04) and object-location binding (p = .03). UFC and DASS-42 scores (all p's > .08) were not significantly changed by either arm of the intervention. No correlations were observed between the change in Lactobacillus and memory performance. Conclusions: Fermented dairy consumption increased the presence of certain microorganisms in the gut and improved relational memory in healthy adults. However, the benefits observed for relational memory were not related to changes in Lactobacillus.Trial registration: ClinicalTrials.gov identifier: NCT02849275.

Dietary lutein plus zeaxanthin and choline intake is interactively associated with cognitive flexibility in middle-adulthood in adults with overweight and obesity.

BACKGROUND: The xanthophyll carotenoids lutein+zeaxanthin and the dietary component choline have been linked to benefits in cognition. However, knowledge on the interactive influence of these dietary components on cognitive function is sparse. DESIGN: 80 middle-aged adults with overweight and obesity (Body Mass Index: (BMI) ≥25.0 kg/m²), completed 7-day diet records, venous blood draws, heterochromatic flicker photometry, assessment of intelligence quotient (IQ), and a cognitive flexibility task while undergoing electroencephalographic recording for event-related potential (ERP) extraction. Multiplicative interaction terms and hierarchical linear regressions, controlling for age, BMI, sex, annual household income, and IQ were utilized to assess independent and interactive contributions of dietary and biomarker data on Switch task outcomes. RESULTS: Higher intake of lutein+zeaxanthin and choline was associated interactively, but not independently, with faster reaction time (RT), after controlling for pertinent covariates. Dietary intake of lutein+zeaxanthin and choline was associated with serum lutein concentrations, but not with plasma choline metabolites nor macular pigmentation. Plasma phosphatidylcholine (PC) concentrations were associated with higher accuracy in Switch trials, while no other biomarkers were associated with cognitive outcomes. Dietary intake and biomarker data were not related to the N2 nor P3 ERP component. CONCLUSIONS: Among a sample of adults with overweight and obesity, greater intake of choline and lutein+zeaxanthin was associated with faster performance on a cognitive flexibility task. Future work examining methods of increasing consumption of both of these dietary components as a possible means of improving or maintaining cognitive flexibility among adults with overweight and obesity is therefore warranted.

Higher protein intake during resistance training does not potentiate strength, but modulates gut microbiota, in middle-aged adults: a randomized control trial.

Protein intake above the recommended dietary allowance (RDA) and resistance training are known anabolic stimuli to support healthy aging. Specifically, protein supplementation after resistance exercise and nightly are strategies to maximize utilization of protein intake above the RDA in healthy adults. As such, the primary objective was to examine the efficacy of protein supplementation and nutritional counseling resulting in either moderate (MOD: ∼1.0 g·kg-1·day-1) or higher (HIGH: ∼1.6 g·kg-1·day-1) protein intake during resistance training on strength (one-repetition maximum, 1-RM; isokinetic and isometric peak torque) in healthy middle-aged adults. Exploratory analyses include diet-exercise effects on lean body mass (LBM), clinical biomarkers, gut microbiota, and diet composition. In all, 50 middle-aged adults (age: 50 ± 8 yr, BMI: 27.2 ± 4.1 kg/m2) were randomized to either MOD or HIGH protein intake during a 10-wk resistance training program (3 × wk). Participants received dietary counseling and consumed either 15 g (MOD) or 30 g (HIGH) of protein from lean beef in the immediate postexercise period and each evening. Maximal strength (1-RM) for all upper and lower body exercises significantly increased with no effect of protein intake (P < 0.050). There was a main effect of time for LBM (P < 0.005). Cardiovascular, renal, or glycemic biomarkers were not affected by the intervention. Gut microbiota were associated with several health outcomes (P < 0.050). In conclusion, higher protein intake above moderate amounts does not potentiate resistance training adaptations in previously untrained middle-aged adults. This trial was registered at clinicaltrials.gov as NCT03029975.NEW & NOTEWORTHY Our research evaluates the efficacy of higher in comparison with moderate animal-based protein intake on resistance exercise training-induced muscle strength, clinical biomarkers, and gut microbiota in middle-aged adults through a dietary counseling-controlled intervention. Higher protein intake did not potentiate training adaptations, nor did the intervention effect disease biomarkers. Both diet and exercise modified gut microbiota composition. Collectively, moderate amounts of high-quality, animal-based protein is sufficient to promote resistance exercise adaptations at the onset of aging.

The relationships between prolonged sedentary time, physical activity, cognitive control, and P3 in adults with overweight and obesity.

BACKGROUND/OBJECTIVES: To assess the relationships between daily sedentary time (ST), prolonged ST, moderate-to-vigorous physical activity (MVPA), and behavioral and neuroelectric indices of cognitive control in adults with overweight and obesity (OW/OB). SUBJECTS/METHODS: A cross-sectional design was used. Overall, 89 adults (BMI = 31.9 ± 4.9 kg/m2) provided measures of ST, prolonged ST (i.e., ST accumulated in ≥20 min), and MVPA from a hip-worn accelerometer worn over 7 days. Inhibitory control was measured with a modified Eriksen flanker task and cognitive flexibility with task switching. The amplitude and the latency of the P3 component of event-related potentials during each task were used as measures of attentional resource allocation and information processing speed, respectively. RESULTS: After adjusting for ST and MVPA, prolonged ST was related to greater interference (i.e., a larger decrement in accuracy between congruent and incongruent trials of the flanker task) indicative of a specific relationship between prolonged ST and poorer inhibitory control. Before adjusting for ST, MVPA was related to a smaller Global Switch Cost expressed as larger (more positive) amplitude of the P3 difference wave (mixed-task minus single-task condition of the switch task). Adjustment for ST attenuated this association to non-significance. CONCLUSIONS: Our findings suggest that future interventions focused on improving inhibitory control in adults with OW/OB should target restructuring ST in addition to current efforts to increase MVPA.

Macular Xanthophylls and Markers of the Anterior Visual Pathway among Persons with Multiple Sclerosis.

BACKGROUND: Multiple sclerosis (MS) can cause retinal thinning among persons with MS with optic neuritis (MS-ON). Macular xanthophylls are carotenoids that comprise the macular pigment, filtering blue light and countering photo-oxidation. However, macular xanthophyll status and its implications for markers of neuroaxonal degeneration have not been examined in MS. OBJECTIVES: This study characterized differences in macular and serum xanthophylls, and retinal morphometry [retinal nerve fiber layer thickness at the macular (mRNFL) and optic disc (odRNFL) and total macular volume (TMV)] in individuals with MS and healthy controls (HC). Associations between macular pigment optical density (MPOD) and retinal morphometry were also examined. METHODS: Adults aged 45-64 y (HC, n = 42; MS, n = 40) participated in a cross-sectional study. MPOD was measured via heterochromatic flicker photometry. Retinal morphometry was measured via optical coherence tomography (OCT). Serum carotenoids were quantified using HPLC. Dietary carotenoids were collected using 7-d records. One-factor ANOVA was conducted to determine group effects on macular, serum, and dietary carotenoids. Partial correlations examined the relations between MPOD, retinal morphometry, diet, and serum carotenoids. RESULTS: Relative to HC, persons with MS-ON had lower MPOD (Cohen's d = 0.84, P = 0.014), lower odRNFL (Cohen's d = 2.16, P <0.001), lower mRNFL (Cohen's d = 0.57, P = 0.028), and lower TMV (Cohen's d = 0.95, P = 0.011). MS without ON (MS) had lower odRNFL (Cohen's d = 0.93, P = 0.001) than HC and lower serum lutein than MS-ON subjects (Cohen's d = 0.65, P = 0.014). Among MS, MPOD was positively correlated with odRNFL thickness (ρ = 0.43, P = 0.049) and TMV (ρ = 0.45, P = 0.039), whereas odRNFL was negatively correlated with serum lutein (ρ = -0.68, P = 0.016) and zeaxanthin (ρ = -0.62, P = 0.028). CONCLUSIONS: Persons with MS-ON exhibited poorer xanthophyll status in the macula and serum. MPOD was associated with beneficial anatomical features in the MS group. These findings warrant confirmation with larger cohorts and prospective trials to evaluate xanthophyll effects on the anterior visual pathway in MS.

Fecal Bacteria as Biomarkers for Predicting Food Intake in Healthy Adults.

BACKGROUND: Diet affects the human gastrointestinal microbiota. Blood and urine samples have been used to determine nutritional biomarkers. However, there is a dearth of knowledge on the utility of fecal biomarkers, including microbes, as biomarkers of food intake. OBJECTIVES: This study aimed to identify a compact set of fecal microbial biomarkers of food intake with high predictive accuracy. METHODS: Data were aggregated from 5 controlled feeding studies in metabolically healthy adults (n = 285; 21-75 y; BMI 19-59 kg/m2; 340 data observations) that studied the impact of specific foods (almonds, avocados, broccoli, walnuts, and whole-grain barley and whole-grain oats) on the human gastrointestinal microbiota. Fecal DNA was sequenced using 16S ribosomal RNA gene sequencing. Marginal screening was performed on all species-level taxa to examine the differences between the 6 foods and their respective controls. The top 20 species were selected and pooled together to predict study food consumption using a random forest model and out-of-bag estimation. The number of taxa was further decreased based on variable importance scores to determine the most compact, yet accurate feature set. RESULTS: Using the change in relative abundance of the 22 taxa remaining after feature selection, the overall model classification accuracy of all 6 foods was 70%. Collapsing barley and oats into 1 grains category increased the model accuracy to 77% with 23 unique taxa. Overall model accuracy was 85% using 15 unique taxa when classifying almonds (76% accurate), avocados (88% accurate), walnuts (72% accurate), and whole grains (96% accurate). Additional statistical validation was conducted to confirm that the model was predictive of specific food intake and not the studies themselves. CONCLUSIONS: Food consumption by healthy adults can be predicted using fecal bacteria as biomarkers. The fecal microbiota may provide useful fidelity measures to ascertain nutrition study compliance.

Avocado Consumption Alters Gastrointestinal Bacteria Abundance and Microbial Metabolite Concentrations among Adults with Overweight or Obesity: A Randomized Controlled Trial.

BACKGROUND: Avocados are rich in dietary fiber and monounsaturated fatty acids (MUFAs), nutrients that have been independently connected to metabolic health benefits and the gastrointestinal microbiota. OBJECTIVES: We aimed to evaluate the impact of avocado consumption on the gastrointestinal microbiota and microbial metabolites, secondary outcomes of the Persea americana for Total Health (PATH) study, and conduct exploratory analyses to assess relations between the fecal microbiota, fecal metabolites, and health markers. METHODS: Adults [n = 163, 25-45 y, BMI (kg/m2) ≥ 25.0] were enrolled in the PATH study, a 12-wk investigator-blinded trial where participants were batch randomized to match the 2 groups by age, sex, visceral adiposity, and fasting glucose concentrations. Participants consumed isocaloric meals with or without avocado (175 g, men; 140 g, women) once daily for 12 wk. The fecal microbiota was assessed with 16S ribosomal RNA gene (V4 region) sequencing and analysis using DADA2 and QIIME2. Fecal fatty acid and bile acid concentrations were quantified using GC and LC-MS. Per-protocol (≥80% meal consumption) and intent-to-treat analyses were conducted using univariate ANOVA and Mann-Whitney U tests. Bivariate correlations were conducted between fecal microbiota, fecal metabolites, and health measures. RESULTS: The avocado treatment increased ɑ diversity and enriched Faecalibacterium, Lachnospira, and Alistipes between 26% and 65% compared with the control group. The avocado group had 18% greater fecal acetate, 70% greater stearic acid, and 98% greater palmitic acid concentrations than the control group, while the concentrations of the bile acids cholic and chenodeoxycholic acid were 91% and 57% lower, respectively. CONCLUSIONS: Daily avocado consumption resulted in lower fecal bile acid concentrations, greater fecal fatty acid and SCFAs, and greater relative abundances of bacteria capable of fiber fermentation, providing evidence that this nutrient-dense food affects digestive physiology, as well as the composition and metabolic functions of the intestinal microbiota. This trial was registered at www.clinicaltrials.gov as NCT02740439.

Avocado Consumption, Abdominal Adiposity, and Oral Glucose Tolerance Among Persons with Overweight and Obesity.

BACKGROUND: Although intake of Hass avocado has been cross-sectionally linked to lower abdominal obesity, knowledge of the effects of avocado consumption on abdominal adiposity and glycemic outcomes remains limited. OBJECTIVE: The effects of avocado consumption on abdominal adiposity, insulin resistance, oral-glucose-tolerance test (OGTT), and estimated ß-cell function were evaluated. METHODS: A total of 105 adults aged 25-45 y (61% female) with BMI ≥25 kg/m2 were randomly assigned to an intervention (N = 53) that received a daily meal with 1 fresh Hass avocado or a control (N = 52) that received an isocaloric meal with similar ingredients without avocado for 12 wk. DXA was used to assess the primary outcomes of abdominal adiposity [visceral adipose tissue (VAT), subcutaneous abdominal adipose tissue (SAAT), and the ratio of VAT to SAAT (VS Ratio)]. Fasted glucose and insulin were used to assess the primary outcomes of insulin resistance (HOMA-IR), and insulin sensitivity (Matsuda index) and ß-cell function (Insulinogenic index) were estimated using an OGTT. Changes between groups were compared using an ANCOVA. Secondary analyses were conducted based on sex. RESULTS: The control group exhibited a greater reduction in SAAT [-54.5 ± 155.8 g (control) compared with 17.4 ± 155.1 g (treatment), P = 0.017] and increase in VS Ratio [0.007 ± 0.047 (control) compared with -0.011 ± 0.044 (treatment), P = 0.024]. Among females, the treatment group exhibited a greater reduction in VAT [1.6 ± 89.8 g (control) compared with -32.9 ± 81.6 g (treatment), P = 0.021] and VS Ratio [0.01 ± 0.05 (control) compared with -0.01 ± 0.03 (treatment), P = 0.001]. Among males, there was no significant difference between groups in changes in abdominal adiposity or glycemic outcomes. CONCLUSIONS: Daily consumption of 1 fresh Hass avocado changed abdominal adiposity distribution among females but did not facilitate improvements in peripheral insulin sensitivity or ß-cell function among adults with overweight and obesity.This study was registered at clinicaltrials.gov as NCT02740439.

Single Nucleotide Polymorphisms in CD36 Are Associated with Macular Pigment among Children.

BACKGROUND: High macular pigment optical density (MPOD) has been associated with improved eye health and better cognitive functions. Genetic variations have been associated with MPOD in adults. However, these associations between genetic variations and MPOD have not been studied in children. OBJECTIVES: This was a secondary analysis of the FK2 (Fitness Improves Thinking in Kids 2) trial (n = 134, 41% male). The aim was to determine differences in MPOD among children (aged 7-9 y) based on genetic variants that either are biologically relevant to lutein (L) and zeaxanthin (Z) accumulation or have been associated with MPOD in adults. METHODS: MPOD was measured using customized heterochromatic flicker photometry via a macular densitometer. DXA was used to assess whole-body and visceral adiposity. DNA was extracted from saliva samples and was genotyped for 26 hypothesis-driven single nucleotide polymorphisms and 75 ancestry-informative markers (AIMs). Habitual diet history was obtained via 3-d food logs completed by parents (n = 88). General linear models were used to compare MPOD between different genotypes. Principal component analysis was performed for the AIMs to account for ethnic heterogeneity. RESULTS: Children carrying ≥1 minor allele on ß-carotene-15,15'-monooxygenase (BCO1)-rs7501331 (T allele) (P = 0.045), cluster of differentiation 36(CD36)-rs1527483 (T allele) (P = 0.038), or CD36-rs3173798 (C allele) (P = 0.001) had significantly lower MPOD (range: 14.1%-26.4%) than those who were homozygotes for the major alleles. MPOD differences based on CD36-rs3173798 genotypes persisted after adjustment for dietary L and Z intake. CONCLUSIONS: The findings indicate that genetic variations of CD36 and BCO1 contribute to MPOD in children. The influence of genetic variation in CD36-rs3173798 persisted after adjusting for variation in dietary intake.This trial was registered at clinicaltrials.gov as NCT01619826.

Dietary choline is related to neural efficiency during a selective attention task among middle-aged adults with overweight and obesity.

Objectives: Obesity is associated with poorer brain health and cognitive function. However, it is not clear whether specific dietary factors may provide neuroprotective effects among individuals with overweight and obesity. The aim of this study was to examine the impact of choline intake on neurophysiological markers of attentional control among young and middle-aged adults with overweight or obesity.Methods: 146 adults with BMI ≥25 kg/m2 (34.0 ± 5.9 years, 57 males) participated in the study. Behavioral performance (accuracy and reaction time) and neuroelectric indices (event-related brain potentials [ERPs]) of attentional inhibition were assessed during a Flanker task. Specifically, the amplitude and latency of the P3 waveform in a central-parietal region of interest (ROI) were used to index attentional resource allocation and information processing speed, respectively. Choline intake and overall diet quality (Healthy Eating Index [HEI-2015]) were assessed using 7-day diet records. Intelligence Quotient was assessed using the Kaufman-Brief Intelligence Test. Regression analyses were conducted to examine the relationship between habitual dietary choline intake and cognitive outcomes following adjustment of demographic factors, IQ, HEI-2015, and BMI.Results: Choline intake was selectively associated with a lower peak amplitude of the P300 waveform during incongruent trials (ß = -0.25, p = <0.01). No significant relationships were observed for accuracy or reaction time.Discussion: Higher choline intake is associated with more efficient neural processing among adults with overweight and obesity. Intervention are necessary to determine whether choline consumption provides neuroprotective effects for executive function among individuals with elevated weight status.