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BACKGROUND: Inadequate dietary fiber (DF) intake is associated with several human diseases. Bread is commonly consumed, and its DF content can be increased by incorporating defatted rice bran (DRB). OBJECTIVE: This first human study on DRB-fortified bread primarily aims to assess the effect of DRB-fortified bread on the relative abundance of a composite of key microbial genera and species in fecal samples. Secondary outcomes include clinical (cardiovascular risk profile), patient-reported (daily bread consumption and bowel movement, gut comfort, general well-being, and total DF intake), biological (fecal microbiota gene abundances, and fecal and plasma metabolites), and physiome (whole-gut and regional transit time and gas fermentation profiles) outcomes in healthy adults with low DF intake. METHODS: This is a 2-armed, placebo-controlled, double-blinded, crossover randomized controlled trial. The study duration is 14 weeks: 2 weeks of lead-in, 4 weeks of intervention per phase, 2 weeks of washout, and 2 weeks of follow-up. Overall, 60 healthy adults with low DF intake (<18 g [female individuals] or <22 g [male individuals] per day) were recruited in Christchurch, New Zealand, between June and December 2022. Randomly assigned participants consumed 3 (female individuals) or 4 (male individuals) slices of DRB-fortified bread per day and then placebo bread, and vice versa. The DRB-fortified bread provided 8 g (female individuals) or 10.6 g (male individuals) of total DF, whereas the placebo (a matched commercial white toast bread) provided 2.7 g (female individuals) or 3.6 g (male individuals) of total DF. Before and after each intervention phase, participants provided fecal and blood samples to assess biological responses; completed a 3-day food diary to assess usual intakes and web-based questionnaires to assess gut comfort, general and mental well-being, daily bread intake, and bowel movement via an app; underwent anthropometry and blood pressure measurements; and drank blue food dye to assess whole-gut transit time. Additionally, 25% (15/60) of the participants ingested Atmo gas-sensing capsules to assess colonic gas fermentation profile and whole-gut and regional transit time. Mean differences from baseline will be compared between the DRB and placebo groups, as well as within groups (after the intervention vs baseline). For metabolome analyses, comparisons will be made within and between groups using postintervention values. RESULTS: Preliminary analysis included 56 participants (n=33, 59% female; n=23, 41% male). Due to the large dataset, data analysis was planned to be fully completed by the last quarter of 2024, with full results expected to be published in peer-reviewed journals by the end of 2024. CONCLUSIONS: This first human study offers insights into the prospect of consuming DRB-fortified bread to effectively modulate health-promoting gut microbes, their metabolism, and DF intake in healthy adults with low DF intake. TRIAL REGISTRATION: Australian New Zealand Clinical Trials Registry ACTRN12622000884707; https://www.anzctr.org.au/Trial/Registration/TrialReview.aspx?id=383814. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): DERR1-10.2196/59227.
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Pan , Estudios Cruzados , Fibras de la Dieta , Alimentos Fortificados , Microbioma Gastrointestinal , Oryza , Humanos , Oryza/química , Fibras de la Dieta/administración & dosificación , Fibras de la Dieta/metabolismo , Masculino , Femenino , Microbioma Gastrointestinal/efectos de los fármacos , Adulto , Método Doble Ciego , Persona de Mediana Edad , Heces/microbiología , Heces/químicaRESUMEN
Previous clinical studies indicate that monoamine oxidase-B (MAO-B) inhibition by blackcurrants must be predominantly attributed to bioactives other than anthocyanins. In this natural products discovery study, MAO-A/B inhibitory phytochemicals were isolated from blackcurrants, and a double-blind crossover study investigated the efficacy of freeze-dried whole-fruit blackcurrant powder in inhibiting MAO-B compared with blackcurrant juice in healthy adults. Platelet MAO-B inhibition was comparable between powder (89% ± 6) and juice (91% ± 4), and it was positively correlated with MAO-modulated plasma catecholamines, subjective alertness, and reduced mental fatigue, assessed using the Bond-Lader questionnaire. Sarmentosin, a nitrile glycoside, and its hydroxycinnamoyl esters were identified as novel MAO-A/B inhibitors from blackcurrant in vitro, and sarmentosin was demonstrated to inhibit platelet MAO-B activity in vivo. These findings confirm sarmentosin as the primary bioactive for MAO-A/B inhibition in blackcurrants, as well as its bioavailability and stability during freeze-drying, and suggest that consuming blackcurrant powder and juice may positively affect mood in healthy adults.
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Plaquetas , Estudios Cruzados , Inhibidores de la Monoaminooxidasa , Monoaminooxidasa , Extractos Vegetales , Ribes , Humanos , Ribes/química , Adulto , Masculino , Monoaminooxidasa/metabolismo , Inhibidores de la Monoaminooxidasa/farmacología , Inhibidores de la Monoaminooxidasa/química , Extractos Vegetales/química , Extractos Vegetales/farmacología , Femenino , Plaquetas/efectos de los fármacos , Plaquetas/enzimología , Plaquetas/metabolismo , Adulto Joven , Método Doble Ciego , Frutas/química , Persona de Mediana EdadRESUMEN
BACKGROUND: The role of gut microbiota in human health has been intensively studied and more recently shifted from emphasis on composition towards function. Function is partly mediated through formed metabolites. Short-chain fatty acids (SCFAs) such as acetate, propionate, and butyrate as well as their branched analogues represent major products from gut fermentation of dietary fibre and proteins, respectively. Robust and high-throughput analysis of SCFAs in small volume blood samples have proven difficult. Major obstacles come from the ubiquitous presence of SCFAs that leads to contaminations and unstable analytical results because of the high volatility of these small molecules. Comprehensive and comparable data on the variation of SCFAs in blood samples from different blood matrices and mammal species including humans is lacking. Therefore, our aim was to develop and evaluate a stable and robust method for quantitation of 8 SCFAs and related fermentation products in small volume blood plasma samples and to investigate their variation in humans and different animal species. RESULTS: Derivatization was a successful approach for measurement of SCFAs in biological samples but quenching of the derivatization reaction was crucial to obtain long-term stability of the derivatized analytes. In total 9 compounds (including succinic acid) were separated in 5 min. The method was linear over the range 0.6-3200 nM formic (FA), acetic (AA), 0.3-1600 nM propionic (PA), and 0.16-800 nM for butyric (BA)-, isobutyric (IBA)-, valeric (VA)-, isovaleric (IVA)-, succinic (SA) and caproic acid (CA). The precision ranged ≤12 % within days and ≤28 % between days (except for CA and VA) in three different plasma quality control (QC) samples (29 batches analyzed over 3 months). The extraction recovery was on average 94 % for the different SCFAs. Typical interquartile range (IQR) concentrations (µM) of SCFAs in human plasma samples were 168 µM (FA), 64 µM (AA), 2.2 µM (PA), 0.54 µM (BA), 0.66 µM (IBA), 0.18 µM (VA), 0.40 µM (IVA), and 0.34 µM (CA). In total, 55 samples per batch/day were successfully analyzed and in total 5380 human plasma samples measured over a 3-year timespan. SIGNIFICANCE: The developed UHPLC-MS based method was suitable for measuring SCFAs in small blood volume samples and enabled robust quantitative data.
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Amino acids are important in several biochemical pathways as precursors to neurotransmitters which impact biological processes previously linked to functional gastrointestinal disorders (FGIDs). Dietary protein consumption, metabolic host processes, and the gut microbiome can influence the plasma concentration of amino acids and neurotransmitters, and their uptake by tissues. The aim of this analysis was to quantify 19 proteogenic and 4 non-proteogenic amino acids and 19 neurotransmitters (including precursors and catabolites, herein referred to as neurotransmitters) to ascertain if their circulating concentrations differed between healthy participants and those with FGIDs. Plasma proteogenic and non-proteogenic amino acids and neurotransmitters were measured using ultra-performance liquid chromatography and liquid chromatography-mass spectrometry, respectively, from 165 participants (Rome IV: irritable bowel syndrome (IBS-constipation, IBS-diarrhea), functional constipation, functional diarrhea, and healthy controls). There were significant differences (p < 0.05) in pairwise comparisons between healthy controls and specific FGID groups for branched-chain amino acids (BCAAs), ornithine, and alpha-aminobutyric acid. No other significant differences were observed for the neurotransmitters or any other amino acids analyzed. Multivariate and bivariate correlation analyses between proteogenic and non-proteogenic amino acids and neurotransmitters for constipation (constipation (IBS-C and functional constipation) and phenotypes diarrhea (IBS-D and functional diarrhea)) and healthy controls suggested that associations between BCAAs, 5-hydroxytryptophan, and kynurenine in combination with tyrosine, 3,4-dihydroxyphenylalanine, and 3,4-dihydroxyphenylacetic acid and associations with gamma-aminobutyric acid, glutamate, asparagine, and serine are likely disrupted in FGID phenotypes. In conclusion, although correlations were evident between some proteogenic and non-proteogenic amino acids and neurotransmitters, the results showed minor concentration differences in plasma proteogenic and non-proteogenic amino acids, amino acid-derived metabolites, and neurotransmitters between FGID phenotypes and healthy controls.
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Plant metabolomics within field-based food production systems is challenging owing to environmental variability and the complex architecture and metabolic growth cycles of plants. Kiwifruit cultivars of Actinidia chinensis are vigorous perennial vines grown as clones in highly structured orchard environments, intensively managed to maximize fruit yield and quality. To understand the metabolic responses of vines to orchard management practices, we needed to better understand the various sources of metabolic variability encountered in the orchard. Triplicate composite leaf, internode and fruit (mature and immature) samples were collected from each of six Actinidia chinensis var. deliciosa 'Hayward' and A. chinensis var. chinensis 'Zesy002' kiwifruit vines at three times during the growing season and measured by LC-MS. In general, there was more variation in metabolite concentrations within vines than between vines, with 'Hayward' showing a greater percentage of within-vine variability than 'Zesy002' (c. 90 vs. 70% respectively). In specific tissues, the sampler, infection by Pseudomonas syringae var. actinidiae and the rootstock also influenced metabolite variability. A similar pattern of metabolic variability was observed from quantitative analysis of specific carbohydrates and phytohormones. High within-vine metabolic variability indicates that it is more important to obtain sufficient replicate samples than to sample from multiple vines. These data provide an objective basis for optimizing metabolite sampling strategies within kiwifruit orchards.
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Whole kiwifruit ('Hayward' and 'Zesy002') were examined for their bioaminergic potential after being subjected to in vitro gastrointestinal digestion and colonic fermentation. Controls included the prebiotic inulin and water, a carbohydrate-free vehicle. The dopamine precursor l-dihydroxyphenylalanine (L-DOPA) and the serotonin precursor 5-hydroxytryptophan were increased in the kiwifruit gastrointestinal digesta ('Hayward' > 'Zesy002') in comparison to the water digesta. Fermentation of the digesta with human fecal bacteria for 18 h modulated the concentrations of bioamine metabolites. The most notable were the significant increases in L-DOPA ('Zesy002' > 'Hayward') and γ-aminobutyric acid (GABA) ('Hayward' > 'Zesy002'). Kiwifruit increased Bifidobacterium spp. and Veillonellaceae (correlating with L-DOPA increase), and Lachnospira spp. (correlating with GABA). The digesta and fermenta were incubated with Caco-2 cells for 3 h followed by gene expression analysis. Effects were seen on genes related to serotonin synthesis/re-uptake/conversion to melatonin, gut tight junction, inflammation and circadian rhythm with different digesta and fermenta from the four treatments. These indicate potential effects of the substrates and the microbially generated organic acid and bioamine metabolites on intestinal functions that have physiological relevance. Further studies are required to confirm the potential bioaminergic effects of gut microbiota-kiwifruit interactions.
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Following cell division, fruit growth is characterized by both expansion through increases in cell volume and biomass accumulation in cells. Fruit growth is limited by carbon starvation; however, the mechanism controlling fruit growth under restricted carbohydrate supply is poorly understood. In a previous study using red-fleshed kiwifruit, we showed that long-term carbon starvation had detrimental effects on carbohydrate, anthocyanin metabolism, and fruit growth. To elucidate the mechanisms underlying the reduction in fruit growth during kiwifruit development, we integrated phytohormone profiling with transcriptomic and developmental datasets for fruit under high or low carbohydrate supplies. Phytohormone profiling of the outer pericarp tissue of kiwifruit showed a 6-fold reduction in total cytokinin concentrations in carbon-starved fruit, whilst other hormones were less affected. Principal component analysis visualised that cytokinin composition was distinct between fruit at 16 weeks after mid bloom, based on their carbohydrate supply status. Cytokinin biosynthetic genes (IPT, CYP735A) were significantly downregulated under carbon starvation, in agreement with the metabolite data. Several genes that code for expansins, proteins involved in cell wall loosening, were also downregulated under carbon starvation. In contrast to other fleshy fruits, our results suggest that cytokinins not only promote cell division, but also drive fruit cell expansion and growth in kiwifruit.
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Emerging evidence from human intervention trials indicates health benefits of consuming blackcurrant fruit, including improvements to cognitive performance, modulation of blood flow, regulation of blood glucose and inhibition of enzymes underpinning normal cognitive function. Of particular relevance is our previous demonstration of monoamine oxidase (MAO)-A and B inhibition after the consumption of a New Zealand "Blackadder" blackcurrant juice in humans. The current study uses a double-blind, placebo-controlled, randomised cross- over design to assess the pharmacodynamics of the effects on platelet MAO-B inhibition and associated substrates, plasma prolactin levels and blood glucose levels after consumption of a single serve of "Blackadder" blackcurrant juice standardised to 500â mg polyphenols. Eight healthy male (20--35 years) participants completed the trial. Measurements were obtained at baseline 15, 30, 45, 60, 100, 120, 150, 180, 240 mins and 24â h post dose. A fast, absolute and reversible inhibition of blood platelet MAO-B (P < 0.001) and a significant but delayed reduction in plasma prolactin (P < 0.001) were observed following the consumption of "Blackadder" blackcurrant juice when compared to a placebo control. No interpretable changes in substrates of MAO or associated metabolites were seen. These data provide a clear time course of the reversible inhibition of MAO-B after the single consumption of a of New Zealand "Blackadder" blackcurrant juice standardised at 500â mg of polyphenols and, therefore, provide a therapeutic window on which to base future nutritional interventions.
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Inhibidores de la Monoaminooxidasa/administración & dosificación , Ácido gammalinolénico/farmacocinética , Adulto , Glucemia/efectos de los fármacos , Plaquetas , Estudios Cruzados , Método Doble Ciego , Humanos , Masculino , Polifenoles/farmacocinética , Prolactina/sangre , Adulto JovenRESUMEN
BACKGROUND: Affective responses experienced during exercise are a significant determinant on exercise adherence. We have previously demonstrated that consumption of New Zealand (NZ) blackcurrants preserves cognition by attenuating the feeling of fatigue. This positive affective response correlated with the ability of blackcurrant polyphenols to support monoamine neurotransmission via inhibition of monoamine oxidase-B (MAO-B) activity. Here we explore how the consumption of a NZ blackcurrant juice (BJ) influenced affective responses and potential ergogenic action on the motivation to adhere to a low impact walking exercise. METHODS: In a parallel randomized controlled study (Trial registration #: ACTRN12617000319370p, registered 28th February 2017, http://www.anzctr.org.au/ ), 40 healthy sedentary male and female participants drank a BJ or matched placebo (PLA) (n = 20 per group), 1 h prior to a self-motivated treadmill walk, where heart rate and affective responses (exertion [ES] or feeling / mood [FS]) scores) were recorded at 3 or 5 min intervals. Blood glucose, lactate, malondialdehyde (MDA) and platelet MAO-B activity were measured pre- and post-exercise and comparisons were conducted using with Student's t-tests. Subjective data were analysed using 2-way ANOVA with appropriate post hoc tests. RESULTS: Consuming a BJ 1 h prior to exercise caused a 90% decline in platelet MAO-B activity. The exercise had no significant (p > 0.05) effect on blood lactate, glucose or plasma MDA levels. Assessment of affective responses over the first 60 mins (adjusting for participant drop-out) revealed a time-dependent ES increase in both groups, with ES reported by participants in the BJ group consistently lower than those in the PLA group (p < 0.05). FS declined in PLA and BJ groups over 60 mins, but an inverse relationship with ES was only observed within the PLA group (r2 = 0.99, p = 0.001). Whilst the average time walked by participants in the BJ group was 11 mins longer than the PLA group (p = 0.3), and 30% of the BJ group achieving > 10 km compared to only 10% for the PLA group (p = 0.28), statistical significance was not achieved. CONCLUSION: Our findings demonstrate that drinking a polyphenolic-rich NZ blackcurrant juice 1 h prior to exercise supports positive affective responses during a self-motivated exercise.
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Afecto , Jugos de Frutas y Vegetales , Motivación , Caminata , Adulto , Glucemia/análisis , Plaquetas/enzimología , Femenino , Frecuencia Cardíaca , Humanos , Ácido Láctico/sangre , Masculino , Malondialdehído/sangre , Monoaminooxidasa/metabolismo , Nueva Zelanda , Polifenoles/administración & dosificación , Ribes , Fenómenos Fisiológicos en la Nutrición Deportiva , Factores de TiempoRESUMEN
We have identified a range of food phytochemicals that inhibit Janus Kinase 2 (JAK2) and Adenosine Monophosphate Kinase (AMPK). A mutated and dysregulated form of JAK2, a tyrosine kinase, is associated with several diseases including Crohn's disease. Using an in vitro, time-resolved fluorescence (TR-FRET) assay, we tested 49 different types of food extracts, plus 10 concentrated fractions of increasing hydrophobicity from each extract, to find foods containing JAK2 inhibitors. The food extracts tested included grains, meat, fish, shellfish, dairy products, herbs, mushrooms, hops, fruits and vegetables. Several fruits were potent inhibitors of JAK2: blackberry, boysenberry, feijoa, pomegranate, rosehip and strawberry, which all contain ellagitannins, known inhibitors of kinases. These fruits are in the Rosales and Myrtales plant orders. No other foods gave >1% of the maximal JAK2 inhibitory activities of these fruits. AMPK, a sensor and regulator of energy metabolism in cells, is a serine-threonine kinase which is reported to be activated by various flavonoid phytochemicals. Using a TR-FRET assay, we tested various fruit extracts for AMPK activation and inhibition. Ellagitannin containing foods scored highly as AMPK inhibitors. Despite several reports of AMPK activation in whole cells by phytochemicals, no extracts or pure compounds activated AMPK in our assay.
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Adenilato Quinasa/antagonistas & inhibidores , Antiinflamatorios no Esteroideos/aislamiento & purificación , Inhibidores Enzimáticos/aislamiento & purificación , Frutas/química , Alimentos Funcionales , Janus Quinasa 2/antagonistas & inhibidores , Fitoquímicos/aislamiento & purificación , Actinidia/química , Adenilato Quinasa/metabolismo , Antiinflamatorios no Esteroideos/química , Antiinflamatorios no Esteroideos/metabolismo , Enfermedad de Crohn/dietoterapia , Enfermedad de Crohn/enzimología , Enfermedad de Crohn/genética , Inhibidores Enzimáticos/química , Inhibidores Enzimáticos/metabolismo , Análisis de los Alimentos , Humanos , Taninos Hidrolizables/aislamiento & purificación , Taninos Hidrolizables/metabolismo , Isoenzimas/antagonistas & inhibidores , Isoenzimas/genética , Isoenzimas/metabolismo , Janus Quinasa 2/genética , Janus Quinasa 2/metabolismo , Lythraceae/química , Mutación , Nueva Zelanda , Fitoquímicos/química , Fitoquímicos/metabolismo , Extractos Vegetales/química , Extractos Vegetales/aislamiento & purificación , Extractos Vegetales/metabolismo , Inhibidores de Proteínas Quinasas/química , Inhibidores de Proteínas Quinasas/aislamiento & purificación , Inhibidores de Proteínas Quinasas/metabolismo , Rosales/químicaRESUMEN
We investigated the biotransformation of four common dietary polyphenols, rutin, quercetin, chlorogenic acid and caffeic acid, in an in vitro mixed culture model of human intestinal microbiota, to determine effects on human gut bacteria. All four compounds were biotransformed rapidly, disappearing from the medium within 0.5 h and later replaced by known phenolic acid breakdown products, at concentrations up to hundreds of micromolar, much higher than in no-polyphenol control experiments. Quantitative PCR was used to measure effects of the polyphenols on the balance between the major groups of intestinal bacteria that are known to influence gut health, i.e., Bifidobacterium spp., Bacteroidetes, and Firmicutes. Fermentation of polyphenols stimulated proliferation of bifidobacteria and decreased the ratio of Firmicutes to Bacteroidetes, relative to controls. Polyphenols also stimulated short chain fatty acid production by the bacteria. Pure bifidobacterial cultures were treated separately with either fermented media isolated from the incubations, the pure test polyphenols, or the biotransformation products detected in the fermentations. Growth stimulation was observed only with fermented polyphenol media and the pure biotransformation products. It appears that dietary polyphenols may have the ability to modify the gut microbial balance, but this effect is indirect, i.e., it is mediated by biotransformation products, rather than the original plant compounds.
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Bacterias/efectos de los fármacos , Biota , Tracto Gastrointestinal/microbiología , Polifenoles/metabolismo , Bacterias/clasificación , Bacterias/genética , Bacterias/crecimiento & desarrollo , Biotransformación , Medios de Cultivo/química , Heces/microbiología , Voluntarios Sanos , Humanos , Masculino , Modelos Teóricos , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
This study was conducted to improve the ability of indigenous New Zealand white-rot fungi to remove pentachlorophenol (PCP) from contaminated field soil. The effects of different bioaugmentation conditions on PCP removal and extracellular enzyme expression were measured in the laboratory. The conditions were fungal growth substrate and co-substrate composition, culture age, and Tween 80 addition to the contaminated soil. The fungi used were Trametes versicolor isolate HR131 and Trametes sp. isolate HR577. Maximum PCP removal was 70% after 7 wk from a 1043 mg kg(-1) PCP-contaminated soil inoculated with an 11-d-old fungal culture of T. versicolor isolate HR131. There was minimal production of undesirable pentachloroanisole by the fungi. Tween 80 addition had no affect on PCP removal. Poplar sawdust was more suitable as a fungal growth substrate and a co-substrate amendment for PCP removal and extracellular enzyme expression than the locally available pine and fir sawdust. Pentachlorophenol removal was not necessarily correlated with extracellular enzyme expression. The research results demonstrate that PCP biodegradation was affected by inoculum culture age, by the presence of a co-substrate amendment, and by growth substrate composition after white-rot fungal bioaugmentation into PCP-contaminated field soils.
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Matriz Extracelular/enzimología , Pentaclorofenol/química , Pentaclorofenol/metabolismo , Peroxidasas/metabolismo , Polyporales/enzimología , Contaminantes del Suelo/metabolismo , beta-Galactosidasa/metabolismo , Nueva Zelanda , Contaminantes del Suelo/química , Factores de TiempoRESUMEN
This study was conducted to improve the pentachlorophenol (PCP) bioremediation ability of white-rot fungi in highly contaminated field soils by manipulating bioaugmentation variables. These were the dry weight percentage of fungal inoculum addition (31-175 g kg(-1)), PCP concentration (100-2137 mg kg(-1) PCP), fungal inoculum formulation, and time (1-7 wk). Five fungal isolates were used: the New Zealand isolates Trametes versicolor (L.: Fr.) HR131 and Trametes sp. HR577; the North American isolates Phanerochaete chrysosporium Burds. (two isolates) and Phanerochaete sordida (Karst.) Erikss. & Ryv. Pentachlorophenol removal, manganese peroxidase, and laccase activity, and the formation of chloroanisoles in the contaminated field soils were measured. The majority of PCP removed by the Trametes isolates was in the first week after bioaugmentation. The maximum PCP removal by the fungi varied from 50 to 65% from a 1065 mg kg(-1) PCP contaminated field soil. Pentachlorophenol was preferentially converted to pentachloroanisole (PCA) by the Phanerochaete isolates (>60%), while 2 to 9% of the PCP removed by two Trametes isolates was converted to PCA. A pH increase was measured following bioaugmentation that was dependent on PCP concentration, fungal inoculum addition, and formulation. This, together with rapid initial PCP removal, possibly changed the bioavailability of the remaining PCP to the fungi and significantly decreased the sequestering of PCP in the contaminated field soils. The research supports the conclusion that New Zealand Trametes spp. can rapidly remove PCP in contaminated field soils. Bioavailability and extractability of PCP in the contaminated field soil may significantly increase after bioaugmentation.