A distinct clade of Bifidobacterium longum in the gut of Bangladeshi children thrives during weaning.

The gut microbiome has an important role in infant health and development. We characterized the fecal microbiome and metabolome of 222 young children in Dhaka, Bangladesh during the first two years of life. A distinct Bifidobacterium longum clade expanded with introduction of solid foods and harbored enzymes for utilizing both breast milk and solid food substrates. The clade was highly prevalent in Bangladesh, present globally (at lower prevalence), and correlated with many other gut taxa and metabolites, indicating an important role in gut ecology. We also found that the B. longum clades and associated metabolites were implicated in childhood diarrhea and early growth, including positive associations between growth measures and B. longum subsp. infantis, indolelactate and N-acetylglutamate. Our data demonstrate geographic, cultural, seasonal, and ecological heterogeneity that should be accounted for when identifying microbiome factors implicated in and potentially benefiting infant development.

A reference map of potential determinants for the human serum metabolome.

The serum metabolome contains a plethora of biomarkers and causative agents of various diseases, some of which are endogenously produced and some that have been taken up from the environment1. The origins of specific compounds are known, including metabolites that are highly heritable2,3, or those that are influenced by the gut microbiome4, by lifestyle choices such as smoking5, or by diet6. However, the key determinants of most metabolites are still poorly understood. Here we measured the levels of 1,251 metabolites in serum samples from a unique and deeply phenotyped healthy human cohort of 491 individuals. We applied machine-learning algorithms to predict metabolite levels in held-out individuals on the basis of host genetics, gut microbiome, clinical parameters, diet, lifestyle and anthropometric measurements, and obtained statistically significant predictions for more than 76% of the profiled metabolites. Diet and microbiome had the strongest predictive power, and each explained hundreds of metabolites-in some cases, explaining more than 50% of the observed variance. We further validated microbiome-related predictions by showing a high replication rate in two geographically independent cohorts7,8 that were not available to us when we trained the algorithms. We used feature attribution analysis9 to reveal specific dietary and bacterial interactions. We further demonstrate that some of these interactions might be causal, as some metabolites that we predicted to be positively associated with bread were found to increase after a randomized clinical trial of bread intervention. Overall, our results reveal potential determinants of more than 800 metabolites, paving the way towards a mechanistic understanding of alterations in metabolites under different conditions and to designing interventions for manipulating the levels of circulating metabolites.

Yoghurt consumption is associated with changes in the composition of the human gut microbiome and metabolome.

BACKGROUND: Yoghurt contains live bacteria that could contribute via modulation of the gut microbiota to its reported beneficial effects such as reduced body weight gain and lower incidence of type 2 diabetes. To date, the association between yoghurt consumption and the composition of the gut microbiota is underexplored. Here we used clinical variables, metabolomics, 16S rRNA and shotgun metagenomic sequencing data collected on over 1000 predominantly female UK twins to define the link between the gut microbiota and yoghurt-associated health benefits. RESULTS: According to food frequency questionnaires (FFQ), 73% of subjects consumed yoghurt. Consumers presented a healthier diet pattern (healthy eating index: beta = 2.17 ± 0.34; P = 2.72x10-10) and improved metabolic health characterised by reduced visceral fat (beta = -28.18 ± 11.71 g; P = 0.01). According to 16S rRNA gene analyses and whole shotgun metagenomic sequencing approach consistent taxonomic variations were observed with yoghurt consumption. More specifically, we identified higher abundance of species used as yoghurt starters Streptococcus thermophilus (beta = 0.41 ± 0.051; P = 6.14x10-12) and sometimes added Bifidobacterium animalis subsp. lactis (beta = 0.30 ± 0.052; P = 1.49x10-8) in the gut of yoghurt consumers. Replication in 1103 volunteers from the LifeLines-DEEP cohort confirmed the increase of S. thermophilus among yoghurt consumers. Using food records collected the day prior to faecal sampling we showed than an increase in these two yoghurt bacteria could be transient. Metabolomics analysis revealed that B. animalis subsp. lactis was associated with 13 faecal metabolites including a 3-hydroxyoctanoic acid, known to be involved in the regulation of gut inflammation. CONCLUSIONS: Yoghurt consumption is associated with reduced visceral fat mass and changes in gut microbiome including transient increase of yoghurt-contained species (i.e. S. thermophilus and B. lactis).

Characterisation, procedures and heritability of acute dietary intake in the Twins UK cohort: an observational study.

BACKGROUND: Estimated food records (EFR) are a common dietary assessment method. This investigation aimed to; (1) define the reporting quality of the EFR, (2) characterise acute dietary intake and eating behaviours, (3) describe diet heritability. METHODS: A total of 1974 one-day EFR were collected from 1858 participants in the TwinsUK cohort between 2012 and 2017. EFR were assessed using a six-point scoring system to determine reporting quality. The frequency and co-occurrence of food items was examined using word clouds and co-occurrence networks. The impact of eating behaviours on weight, BMI and nutrient intake were explored using mixed-effect linear regression models. Finally, diet heritability was estimated using ACE modelling. RESULTS: We observed that 75% of EFR are of acceptable reporting quality (score > 5). Black tea and semi-skimmed milk were the most consumed items, on an individual basis (respectively 8.27, 6.25%) and paired (0.21%) as co-occurring items. Breakfast consumption had a significantly (p = 5.99 × 10- 7) greater impact on energy (kcal) (mean 1874.67 (±SD 532.42)) than skipping breakfast (1700.45 (±SD 620.98)), however only length of eating window was significantly associated with body weight (kg) (effect size 0.21 (±SD 0.10), p = 0.05) and BMI (effect size 0.08 (±SD 0.04), p = 0.04) after adjustment for relevant covariates. Lastly, we reported that both length of eating window (h2 = 33%, CI 0.24; 0.41), and breakfast consumption (h2 = 11%, CI 0.02; 0.21) were weakly heritable. CONCLUSIONS: EFR describing acute dietary intake allow for eating behaviour characterisation and can supplement habitual diet intake assessments. Novel findings of heritability warrant further investigation.

Self-Reported Symptoms of COVID-19, Including Symptoms Most Predictive of SARS-CoV-2 Infection, Are Heritable.

Susceptibility to infection such as SARS-CoV-2 may be influenced by host genotype. TwinsUK volunteers (n = 3261) completing the C-19 COVID-19 symptom tracker app allowed classical twin studies of COVID-19 symptoms, including predicted COVID-19, a symptom-based algorithm to predict true infection, derived from app users tested for SARS-CoV-2. We found heritability of 49% (32-64%) for delirium; 34% (20-47%) for diarrhea; 31% (8-52%) for fatigue; 19% (0-38%) for anosmia; 46% (31-60%) for skipped meals and 31% (11-48%) for predicted COVID-19. Heritability estimates were not affected by cohabiting or by social deprivation. The results suggest the importance of host genetics in the risk of clinical manifestations of COVID-19 and provide grounds for planning genome-wide association studies to establish specific genes involved in viral infectivity and the host immune response.

Antibiotic treatment triggers gut dysbiosis and modulates metabolism in a chicken model of gastro-intestinal infection.

BACKGROUND: Infection of the digestive track by gastro-intestinal pathogens results in the development of symptoms ranging from mild diarrhea to more severe clinical signs such as dysentery, severe dehydration and potentially death. Although, antibiotics are efficient to tackle infections, they also trigger dysbiosis that has been suggested to result in variation in weight gain in animal production systems. RESULTS: Here is the first study demonstrating the metabolic impact of infection by a gastro-intestinal pathogen (Brachyspira pilosicoli) and its resolution by antibiotic treatment (tiamulin) on the host (chicken) systemic metabolism and gut microbiota composition using high-resolution 1H nuclear magnetic resonance (NMR) spectroscopy and 16S rDNA next generation sequencing (NGS). Clear systemic metabolic markers of infections such as glycerol and betaine were identified. Weight loss in untreated animals was in part explained by the observation of a modification of systemic host energy metabolism characterized by the utilization of glycerol as a glucose precursor. However, antibiotic treatment triggered an increased VLDL/HDL ratio in plasma that may contribute to reducing weight loss observed in treated birds. All metabolic responses co-occurred with significant shift of the microbiota upon infection or antibiotic treatment. CONCLUSION: This study indicates that infection and antibiotic treatment trigger dysbiosis that may impact host systemic energy metabolism and cause phenotypic and health modifications.

Metabonomics-based analysis of Brachyspira pilosicoli's response to tiamulin reveals metabolic activity despite significant growth inhibition.

Pathogenic anaerobes Brachyspira spp. are responsible for an increasing number of Intestinal Spirochaetosis (IS) cases in livestock against which few approved treatments are available. Tiamulin is used to treat swine dysentery caused by Brachyspira spp. and recently has been used to handle avian intestinal spirochaetosis (AIS). The therapeutic dose used in chickens requires further evaluation since cases of bacterial resistance to tiamulin have been reported. In this study, we evaluated the impact of tiamulin at varying concentrations on the metabolism of B. pilosicoli using a 1H-NMR-based metabonomics approach allowing the capture of the overall bacterial metabolic response to antibiotic treatment. Based on growth curve studies, tiamulin impacted bacterial growth even at very low concentration (0.008 µg/mL) although its metabolic activity was barely affected 72 h post exposure to antibiotic treatment. Only the highest dose of tiamulin tested (0.250 µg/mL) caused a major metabolic shift. Results showed that below this concentration, bacteria could maintain a normal metabolic trajectory despite significant growth inhibition by the antibiotic, which may contribute to disease reemergence post antibiotic treatment. Indeed, we confirmed that B. pilosicoli remained viable even after exposition to the highest antibiotic dose. This paper stresses the need to ensure new evaluation of bacterial viability post bacteriostatic exposure such as tiamulin to guarantee treatment efficacy and decrease antibiotic resistance development.

Brachyspira pilosicoli-induced avian intestinal spirochaetosis.

Avian intestinal spirochaetosis (AIS) is a common disease occurring in poultry that can be caused by Brachyspira pilosicoli, a Gram-negative bacterium of the order Spirochaetes. During AIS, this opportunistic pathogen colonises the lower gastrointestinal (GI) tract of poultry (principally, the ileum, caeca, and colon), which can cause symptoms such as diarrhoea, reduced growth rate, and reduced egg production and quality. Due to the large increase of bacterial resistance to antibiotic treatment, the European Union banned in 2006 the prophylactic use of antibiotics as growth promoters in livestock. Consequently, the number of outbreaks of AIS has dramatically increased in the UK resulting in significant economic losses. This review summarises the current knowledge about AIS infection caused by B. pilosicoli and discusses various treatments and prevention strategies to control AIS.

Bifunctional epitope-agonist ligands of the bradykinin B(2) receptor.

Two bradykinin (BK) B(2) receptor agonists N-terminally extended with the myc epitope were synthesized and evaluated: myc-KPG-BK and myc-KGP-B-9972. The latter was modeled on the inactivation-resistant agonist B-9972 (D-Arg(0), Hyp(3), Igl(5), Oic(7), Igl(8)-BK) and is also resistant to endosomal inactivation. Despite a large loss of affinity relative to the parent peptide, the tagged analogs are conventional agonists in the umbilical vein contractility assay and compete for [(3)H]BK binding at the rabbit B(2) receptor. Endocytosed myc-KGP-B-9972 most effectively carried AlexaFluor-488-conjugated anti-myc monoclonal antibodies into intact cells expressing the B(2) receptor. Results support the prospects of functionally-active cargoes entering cells in a pharmacologically controlled manner.

High affinity capture and concentration of quinacrine in polymorphonuclear neutrophils via vacuolar ATPase-mediated ion trapping: comparison with other peripheral blood leukocytes and implications for the distribution of cationic drugs.

Many cationic drugs are concentrated in acidic cell compartments due to low retro-diffusion of the protonated molecule (ion trapping), with an ensuing vacuolar and autophagic cytopathology. In solid tissues, there is evidence that phagocytic cells, e.g., histiocytes, preferentially concentrate cationic drugs. We hypothesized that peripheral blood leukocytes could differentially take up a fluorescent model cation, quinacrine, depending on their phagocytic competence. Quinacrine transport parameters were determined in purified or total leukocyte suspensions at 37 °C. Purified polymorphonuclear leukocytes (PMNLs, essentially neutrophils) exhibited a quinacrine uptake velocity inferior to that of lymphocytes, but a consistently higher affinity (apparent KM 1.1 vs. 6.3 µM, respectively). However, the vacuolar (V)-ATPase inhibitor bafilomycin A1 prevented quinacrine transport or initiated its release in either cell type. PMNLs capture most of the quinacrine added at low concentrations to fresh peripheral blood leukocytes compared with lymphocytes and monocytes (cytofluorometry). Accumulation of the autophagy marker LC3-II occurred rapidly and at low drug concentrations in quinacrine-treated PMNLs (significant at ≥2.5 µM, ≥2 h). Lymphocytes contained more LAMP1 than PMNLs, suggesting that the mass of lysosomes and late endosomes is a determinant of quinacrine uptake Vmax. PMNLs, however, exhibited the highest capacity for pinocytosis (uptake of fluorescent dextran into endosomes). The selectivity of quinacrine distribution in peripheral blood leukocytes may be determined by the collaboration of a non-concentrating plasma membrane transport mechanism, tentatively identified as pinocytosis in PMNLs, with V-ATPase-mediated concentration. Intracellular reservoirs of cationic drugs are a potential source of toxicity (e.g., loss of lysosomal function in phagocytes).

Inhibitory effects of cytoskeleton disrupting drugs and GDP-locked Rab mutants on bradykinin B2 receptor cycling.

The bradykinin (BK) B2 receptor (B2R) is G protein coupled and phosphorylated upon agonist stimulation; its endocytosis and recycling are documented. We assessed the effect of drugs that affect the cytoskeleton on B2R cycling. These drugs were targeted to tubulin (paclitaxel, or the novel combretastatin A-4 mimetic 3,4,5-trimethoxyphenyl-4-(2-oxoimidazolidin-1-yl)benzenesulfonate [IMZ-602]) and actin (cytochalasin D). Tubulin ligands did not alter agonist-induced receptor endocytosis, as shown using antibodies reactive with myc-tagged B2Rs (microscopy, cytofluorometry), but rather reduced the progression of the ligand-receptor-ß-arrestin complex from the cell periphery to the interior. The 3 fluorescent probes of this complex (B2R-green fluorescent protein [B2R-GFP], the fluorescent agonist fluorescein-5-thiocarbamoyl-D-Arg-[Hyp³, Igl5, Oic7, Igl8]-BK and ß-arrestin2-GFP) were condensed in punctuate structures that remained close to the cell surface in the presence of IMZ-602. Cytochalasin D selectively inhibited the recycling of endocytosed B2R-GFP (B2R-GFP imaging, [³H]BK binding). Dominant negative (GDP-locked)-Rab5 and -Rab11 reproduced the effects of inhibitors of tubulin and actin, respectively, on the cycling of B2R-GFP. GDP-locked-Rab4 also inhibited B2R-GFP recycling to the cell surface. Consistent with the displacement of cargo along specific cytoskeletal elements, Rab5-associated progression of the endocytosed BK B2R follows microtubules toward their (-) end, while its recycling progresses along actin fibers to the cell surface. However, tubulin ligands do not suppress the tested desensitization or resensitization mechanisms of the B2R.

Defining and Directing Gut Microbiota Maturation in Early Life: A Nutrition Perspective.

The development of the microbiome within the human digestive tract starts at birth and continues up to approximately 3 years of age when the microbial ecosystem resembles a more adulthood-like state. The pace of colonization and diversification of the gut microbiota in the early stages of life has been linked to short- and long-term health outcomes. Characterizing optimal maturation of the ecosystem may help identifying adverse events that impair the process and also factors that support and guide it, such as diet. To date, researchers have looked at the evolution over time of gut microbiota parameters such as diversity, taxa abundance, or specific functions. A more global approach has used "microbiota age" to capture maturation trajectory through machine learning models. In this review, the use and limitations of the latest methods to capture and understand microbiota maturation will be discussed. Then the role of nutrition in directing gut microbiota maturation in early life will be described together with the challenges that limit our comprehension of the effects of diet on the gut microbiota.

Assessment of the Feasibility of Converting the Liquid Fraction Separated from Fruit and Vegetable Waste in a UASB Digester.

Anaerobic digestion of food waste still faces important challenges despite its world-wide application. An important fraction of food waste is composed of organic material having a low hydrolysis rate and which is often not degraded in digesters. The addition of this less hydrolysable fraction into anaerobic digesters requires a longer hydraulic residence time, and therefore leads to oversizing of the digesters. To overcome this problem, the conversion of the highly biodegradable liquid fraction from fruit and vegetable waste in a up-flow anaerobic sludge blanket (UASB) digester is proposed and demonstrated. The more easily biodegradable fraction of the waste is concentrated in the liquid phase using a 2-stage screw press separation. Then, this liquid fraction is digested in a 3.5 L UASB digester at a high organic loading rate. A good and stable performance was observed up to an organic loading rate (OLR) of 12 g COD/(Lrx.d), with a specific methane production of 2.6 L CH4/(Lrx.d) and a degradation of 85% of the initial total COD. Compared to the conversion of the same initial waste with a continuously stirred tank reactor (CSTR), this new treatment strategy leads to 10% lower COD degradation, but can produce the same amount of methane with a digester that is twice as small. The scale-up of this process could contribute to reduced costs related to the anaerobic digestion of food waste, while reducing management efforts associated with digestate handling and increasing process stability at high organic loading rates.

Effects of dietary zinc/copper ratios on the metabolism of zinc and copper in weaned pigs.

This study compared the effects of different dietary zinc/copper ratios on zinc (Zn) and copper (Cu) metabolism in weaned pigs. One hundred and sixty piglets (7.81 ±â€…0.25 kg; 21 d of age) were used in a completely randomized 2 × 2 factorial design composed with high (H) and low (L) levels of added dietary Zn (100 and 3,000 mg/kg) and dietary Cu (6 and 130 mg/kg). Piglets were slaughtered at 21, 28, 35, and 42 d of age for blood and tissues collection. Serum, jejunum mucosa, liver, and kidney concentrations of Zn and Cu were analyzed as well as tissues mRNA abundance of genes related to their metabolism. Serum and liver Zn concentrations increased at days 28, 35, and 42 in HZn groups compared to pre-treatment levels (day 21; P ≤ 0.01) but for LZn animals, values decreased at days 28, 35, and 42 in liver (P ≤ 0.01) but remained stable vs. day 21 levels in serum (P ≥ 0.37). Serum, jejunum mucosa, liver, and kidney Zn concentrations were greater in HZn groups from day 28 (P ≤ 0.01). In jejunum mucosa, the mRNA expression of ZIP4 was lower in HZn piglets at day 28 (P ≤ 0.01) and at day 42 whereas HCu supplementation increased ZIP4 expression in LZn but not in HZn diets (P = 0.05). For ZNT1, MT3, and MT1, values of relative mRNA expression were greater for HZn animals in jejunum mucosa, liver, and kidney (P ≤ 0.01) from day 28. In kidney (P < 0.01) at day 42, HZn supplementation increased MTs expression in both LCu or HCu groups. Serum and liver Cu concentrations decreased at days 35 and 42 in all treatments compared to day 21 (P ≤ 0.04), except LZnHCu in liver that was not different from day 21 (P ≥ 0.17). Serum Cu concentrations were lower in HZn and greater in HCu groups at days 35 and 42 (P ≤ 0.01) whereas hepatic Cu was reduced by HZn diets in both LCu and HCu groups at days 35 and 42 (P ≤ 0.01). Jejunum Cu concentrations were increased by HCu diets in HZn but not in LZn groups at days 28 and 42 (P ≤ 0.04). Renal Cu concentrations were greater in HZn groups at day 28 (P < 0.01) whereas at day 42 HZn diets increased Cu values in both LCu and HCu groups (P ≤ 0.01). The expression of ATP7A in kidney at day 42 was greater for HZn groups (P = 0.02). In conclusion, high dietary Zn levels were not efficiently regulated by homeostatic mechanisms and significantly impaired Cu homeostasis. Low dietary Zn/Cu ratios allow a more efficient regulation of the metabolism of these trace minerals in post-weaning piglets. The current official recommendations for Zn and Cu to post-weaning piglets apparently do not fulfill their requirements.

Zinc oxide and copper sulfate are commonly used as growth promoters and alternatives to antibiotics to prevent diarrhea in weaned piglets but their use in post-weaning pigs diets has been challenged due to environmental issues and concerns related to bacterial resistance to antibiotics and heavy metals. Recently, it was reported that high dietary zinc levels interfere with copper status and may be detrimental to post-weaning piglets' health. In fact, the optimal dietary zinc/copper ratios need to be determined. Therefore, this experiment was conducted to evaluate the effects of different dietary zinc/copper ratios (3,000/130, 3,000/6, 100/130, and 100/6 mg/kg) on zinc and copper metabolism in weaned piglets. This study demonstrated that high dietary zinc/copper ratios impaired zinc and copper homeostasis but also that 100 mg/kg of dietary zinc and 6 mg/kg of dietary copper are apparently not sufficient to fulfill the piglets' requirements during the first weeks post-weaning.

Effects of dietary zinc oxide levels on the metabolism of zinc and copper in weaned pigs.

This study compared different dietary zinc oxide (ZnO) levels on zinc (Zn) and copper (Cu) metabolism in weaned pigs. One hundred twenty weaned piglets (7.96 ± 1.17 kg; 21 d of age) were used in a completely randomized 3 × 4 factorial design composed with three levels of dietary ZnO at 100 (100Zn), 1,000 (1,000Zn), or 3,000 mg/kg (3,000Zn) and four ages at slaughter at 21 (day 21), 23 (day 23), 35 (day 35), and 42 d (day 42). Dietary Cu levels were constant at 130 mg/kg. Serum, jejunum, liver, and kidney levels of Zn and Cu as well as mRNA abundance of genes related to Zn and Cu metabolism were analyzed. Zinc levels were greatest in 3,000Zn piglets from day 35 in all tissues (P ≤ 0.01). In 3,000Zn piglets, mRNA expression of ZIP4 was reduced in jejunum whereas ZnT1 and MT3 were stimulated in jejunum and liver and MT1 in kidney (P ≤ 0.04) from day 35. Copper levels were greatest in jejunum (P = 0.06) and kidney (P ≤ 0.01; days 35 and 42 only) and lowest in liver and serum (P ≤ 0.01) of 3,000Zn piglets. In conclusion, the treatment containing 3,000 mg ZnO/kg triggered Zn homeostatic mechanisms in weaned pigs and impaired Cu metabolism through high enterocyte and kidney Cu sequestration.

Zinc oxide (ZnO) is commonly used in post-weaning pig diets as growth promoter alternative to antibiotics to prevent diarrhea. The use of supranutritional levels of ZnO in post-weaning pigs diets has been challenged due to environmental issues and concerns related to bacterial resistance to antibiotics and heavy metals. However, the limited knowledge of the consequences of high levels of dietary ZnO on the metabolism of trace minerals has hampered advances to replace this nutritional strategy without compromising piglets health. Therefore, this experiment was conducted to evaluate the effects of increasing levels of dietary ZnO (i.e., 100, 1,000, and 3,000 mg/kg) on Zn and Cu metabolism in weaned piglets. In this experiment, it was demonstrated that systemic Zn levels were not effectively regulated with supplementation levels at 3,000 mg of ZnO/kg of diet. In addition, this level of dietary ZnO increased the intestinal intracellular sequestration of Cu and impaired its renal reabsorption, negatively impacting hepatic, and systemic serum Cu concentrations. These results emphasize the potential risk of Cu deficiency under long-term supranutritional supplementation of dietary ZnO during the post-weaning period, with potentially detrimental impacts on piglets growth.

Application of Computational Data Modeling to a Large-Scale Population Cohort Assists the Discovery of Inositol as a Strain-Specific Substrate for Faecalibacterium prausnitzii.

Faecalibacterium prausnitzii (F. prausnitzii) is a bacterial taxon in the human gut with anti-inflammatory properties, and this may contribute to the beneficial effects of healthy eating habits. However, little is known about the nutrients that enhance the growth of F. prausnitzii other than simple sugars and fibers. Here, we combined dietary and microbiome data from the American Gut Project (AGP) to identify nutrients that may be linked to the relative abundance of F. prausnitzii. Using a machine learning approach in combination with univariate analyses, we identified that sugar alcohols, carbocyclic sugar, and vitamins may contribute to F. prausnitzii growth. We next explored the effects of these nutrients on the growth of two F. prausnitzii strains in vitro and observed robust and strain-dependent growth patterns on sorbitol and inositol, respectively. In the context of a complex community using in vitro fermentation, neither inositol alone nor in combinations with vitamin B exerted a significant growth-promoting effect on F. prausnitzii, partly due to high variability among the fecal microbiota community from four healthy donors. However, the fecal communities that showed an increase in F. prausnitzii on inulin also responded with at least 60% more F. prausnitzii on any of inositol containing media than control. Future nutritional studies aiming to increase the relative abundance of F. prausnitzii should explore a personalized approach accounting for strain-level genetic variations and community-level microbiome composition.

Interplay between the (Poly)phenol Metabolome, Gut Microbiome, and Cardiovascular Health in Women: A Cross-Sectional Study from the TwinsUK Cohort.

BACKGROUND: Dietary (poly)phenol consumption is inversely associated with cardiovascular disease (CVD) risk in epidemiological studies, but little is known about the role of the gut microbiome in this relationship. METHODS: In 200 healthy females, aged 62.0 ± 10.0 years, from the TwinsUK cohort, 114 individual (poly)phenol metabolites were measured from spot urine using ultra-high-performance liquid chromatography-mass spectrometry. The associations between metabolites, the gut microbiome (alpha diversity and genera), and cardiovascular scores were investigated using linear mixed models adjusting age, BMI, fibre, energy intake, family relatedness, and multiple testing (FDR < 0.1). RESULTS: Significant associations were found between phenolic acid metabolites, CVD risk, and the gut microbiome. A total of 35 phenolic acid metabolites were associated with the Firmicutes phylum, while 5 metabolites were associated with alpha diversity (FDR-adjusted p < 0.05). Negative associations were observed between the atherosclerotic CVD (ASCVD) risk score and five phenolic acid metabolites, two tyrosol metabolites, and daidzein with stdBeta (95% (CI)) ranging from -0.05 (-0.09, -0.01) for 3-(2,4-dihydroxyphenyl)propanoic acid to -0.04 (-0.08, -0.003) for 2-hydroxycinnamic acid (FDR-adjusted p < 0.1). The genus 5-7N15 in the Bacteroidetes phylum was positively associated with the same metabolites, including 3-(3,5-dihydroxyphenyl)propanoic acid, 3-(2,4-dihydroxyphenyl)propanoic acid, 3-(3,4-dihydroxyphenyl)propanoic acid), 3-hydroxyphenylethanol-4-sulfate, and 4-hydroxyphenylethanol-3-sulfate)(stdBeta (95% CI): 0.23 (0.09, 0.36) to 0.28 (0.15, 0.42), FDR-adjusted p < 0.05), and negatively associated with the ASCVD score (stdBeta (95% CI): -0.05 (-0.09, -0.01), FDR-adjusted p = 0.02). Mediation analysis showed that genus 5-7N15 mediated 23.8% of the total effect of 3-(3,4-dihydroxyphenyl)propanoic acid on the ASCVD score. CONCLUSIONS: Coffee, tea, red wine, and several vegetables and fruits, especially berries, are the most abundant food sources of phenolic acids that have the strongest associations with CVD risk. We found that the gut microbiome, particularly the genus 5-7N15, partially mediates the negative association between urinary (poly)phenols and cardiovascular risk, supporting a key role of the gut microbiome in the health benefits of dietary (poly)phenols.

Prolonged signalling and trafficking of the bradykinin B2 receptor stimulated with the amphibian peptide maximakinin: insight into the endosomal inactivation of kinins.

Maximakinin, a 19-residue peptide from the amphibian Bombina maxima, incorporates the full sequence of bradykinin (BK) at its C-terminus with a hydrophilic 10-residue N-terminal extension. As a putative venom component, it may stimulate BK B(2) receptors (B(2)Rs) in a distinct manner relative to the fragile mammalian agonist BK. Maximakinin affinity for B(2)Rs and angiotensin converting enzyme (ACE) and its pharmacological profile have been compared to those of BK. Maximakinin is an agonist of the human and rabbit B(2)R with a 8-12 fold lesser potency, but a prolonged duration of action relative to BK (ERK MAP kinase activation, c-Fos induction in HEK 293 cells). Maximakinin had a moderately inferior affinity (∼6-fold vs. BK) for recombinant ACE based on [(3)H]enalaprilat binding displacement. Unlike BK, maximakinin induced the internalization of the fusion protein B(2)R-green fluorescent protein (GFP) and the downregulation of this construction over a 12-h stimulation period, reproducing the effect of inactivation-resistant B(2)R agonists. Alternate homologues of BK extended at the N-terminus showed intermediate behaviours between BK and maximakinin in the B(2)R-GFP downregulation assay. The recycling of B(2)R-GFP at the cell surface after a 3-h BK treatment was notably inhibited by cotreatment with E-64 or bafilomycin A1, supporting that an endosomal cysteine protease degrades kinins in a process that determines the cycling and fate of the B(2)R. Maximakinin is the first known natural kinin sequence that elicits a prolonged cellular signalling, thus suggesting a possible basis for a venomous action and a naturally selected one for the design of B(2)R-transported biotechnological cargoes.