Food & Function

Background: There is a growing availability of plant-based alternatives for bovine milk. However, plant-based drinks differ e.g. in protein quantity, protein quality and carbohydrate composition. Knowledge on the impact this has on digestion-related outcomes such as postprandial amino acid (AA) availability and gastric behavior is still limited. Additionally, data on milk alternatives with higher overall protein concentration are lacking. Objective: We aimed to compare the differences in postprandial AA profiles, glycemic response, and gastric behavior between bovine milk and plant-based drinks with or without added protein using representative commercial drinks. Methods: 12 healthy males participated in a randomized crossover study with three treatments. After an overnight fast, participants drank 750 ml of semi-skimmed milk (Milk), a standard oat-based drink (OBD) or an oat-based drink enriched with pea protein (OBD+Pea). Blood samples were taken for measurement of plasma AA, glucose, insulin and FGF21 over 5 hours. Gastric content changes were assessed using magnetic resonance imaging scans for Milk and OBD+Pea over 2 hours. Results: Compared with Milk, both oat-based drinks elicited lower 5-h total AA iAUC (OBD -169706; OBD+pea -139552 microM*min; both P<0.001) and EAA iAUC (OBD -91748; OBD+pea -78199; both P<0.001), with no differences between oat-based drinks. Gastric content volume was lower for OBD+Pea versus Milk from 60-120 min (range -48 - -84 ml, all P<0.001). Compared to Milk, OBD and OBD+Pea increased glucose more from 15-45 min (range 0.86 - 1.73 mmol/l, all P<0.001) and insulin from 30-60 min (1.84 - 2.14-fold, all P<0.001). Conclusions: Protein enrichment in plant-based milk alternatives can be a valuable strategy to increase protein content and improve amino acid composition. However, such products still show faster gastric emptying, lower postprandial essential amino acid and higher glycemic and insulinemic responses compared to bovine milk because of their different carbohydrate and protein content and quality.

Traditional Mongolian fermented foods have been extensively utilized for dietary regulation and the promotion of gastrointestinal health. However, spontaneous fermentation remains inherently unpredictable, leading to significant variations in microbial community dynamics, metabolite accumulation, and the consistency and quality of the final product. Drawing on the traditional preparation of Mongolian acidic foods, this study established a controlled production strategy for whole-wheat probiotic fermented soup (WWPFS) by combining enzymatic pretreatment with probiotic-directed fermentation. Physicochemical characterization, 16S rRNA gene-based microbial community profiling, LC-MS/MS-based untargeted metabolomics, safety evaluation, and an Escherichia coli-induced gut microbiota dysbiosis model were employed to optimize and comprehensively characterize the fermentation process of WWPFS. The optimized process established a reproducible fermentation system consistently dominated by Lactobacillus and Bacillus across independent fermentation batches. Compared with traditional spontaneous fermentation, probiotic-directed fermentation remodeled the physicochemical properties of the whole-wheat matrix, including carbon, nitrogen, phosphorus, sulfur, and mineral composition, and facilitated the accumulation of putatively annotated LC-MS/MS features, including DL-lactate, 1,4-D-xylobiose, diacetyl, and phenyllactic-acid-related features derivatives. Acute oral and 28-day repeated-dose toxicity evaluations showed no treatment-related adverse effects within the tested dose range and study duration. In the Escherichia coli-induced gut microbiota dysbiosis mouse model, microbial richness, diversity, and community structure differed among the experimental groups, and both low- and high-dose WWPFS groups showed significant shifts in overall gut microbial community composition relative to the model group after multiple-testing correction, together with directional recovery of selected model-responsive bacterial genera. Cross-system integration identified coordinated response patterns between fermentation-derived metabolite features and model-responsive gut bacterial taxa, supporting a potential metabolite-microbiota link in WWPFS-mediated gut microbiota modulation. In summary, probiotic-directed fermentation improved the controllability of the traditional Mongolian fermented food production process, reshaped its metabolic profile, and enhanced its potential to modulate the gut microbiota. These findings provide experimental evidence supporting the modernization of traditional Mongolian fermented foods and the development of probiotic-based functional foods.

BackgroundObesity is a complex multifactorial disease associated with chronic low grade inflammation, gut microbiota dysbiosis, and impaired gut-brain communication. Oligomeric procyanidins from grape seed extracts (GSE) are promising prebiotic candidates, capable of modulating host metabolism through interactions with the gut microbiota. MethodsC57Bl/6J male mice were rendered obese by feeding them a high fat, high sucrose diet and were orally administered GSE at a dose of 1or 2 g/kg/day for 12 weeks. We assessed body weight, adiposity, glucose tolerance, insulin sensitivity, circulating hormones, brain homeostasis markers, colonic and liver gene expression, 16S rRNA gene sequencing of the gut microbiota profiles, and untargeted cecal metabolomics. ResultsGSE reduced body weight gain, visceral adiposity, adipocyte hypertrophy, and improved oral glucose tolerance and insulin sensitivity. It normalized circulating lipid and glucose levels and lowered fasting insulin and leptin while increasing endogenous GLP-1. Hepatic gene expression analysis revealed a dose-dependent restoration of antioxidant defenses (SOD, CAT) and lipogenic transcription factors (SREBP, ChREBP). In the colon, GSE attenuated pro-inflammatory IL6 cytokine expression and strikingly upregulated GLP-1 and GLP-1 receptor expression. Microbiota analysis revealed a profound, dose-dependent remodeling of gut microbiota composition and diversity, with an expansion of health-associated taxa, such as Akkermansia muciniphila. Brain analyses revealed restoration of NAA and BDNF levels together with markers consistent with improved mitochondrial function. Cecal metabolomics revealed normalization of secondary bile acid metabolism, restoration of arginine bioavailability, and reduction in the accumulation of L-DOPA and spermidine. ConclusionsAn oligomeric procyanidin-rich grape seed extract acts as a multitarget prebiotic that alleviates diet-induced obesity and is associated with coordinated restoration of gut microbiota composition, GLP-1 signaling, and gut-brain and gut-liver communication pathways. Convergent dose-dependent effects on Akkermansia muciniphila abundance, GLP-1, NAA, and BDNF identify key mechanisms underlying its metabolic benefits.

Phosphatidylserine (PS) is a functional phospholipid widely studied for cognitive support, yet evidence in healthy adults with subjective cognitive decline remains limited. This 12-week randomized, double-blind, placebo-controlled trial evaluated the safety and cognitive effects of 300 mg/day of soybean-derived PS in 70 adults aged 35-65 years reporting subjective memory complaints. PS was well tolerated, with no adverse events or clinically significant changes in safety parameters. In the prespecified primary analysis, the primary endpoint of composite memory score did not differ significantly between groups at week 12. Exploratory secondary analyses suggested possible domain-specific effects, including favorable changes in working memory (p = 0.037) and motor speed (p = 0.005), with the latter appearing to vary according to blood fatty-acid profiles. These findings should be interpreted cautiously and require confirmation in future studies designed to evaluate these domains and potential nutritional moderators.

Inflammatory bowel disease challenges modern gastroenterology due to high relapse rates. This study evaluated the effects of medicinal fungal sclerotium Fu Ling (Wolfiporia cocos), its derived {beta}-1,3-1,6-Glucan, Bacillus coagulans GBI-30, 6086, and their combinations in dextran sulfate sodium-induced colitis in male BALB/c mice. The {beta}-glucan-B. coagulans combination significantly improved body weight, disease activity index, colon length and architecture, mucus secretion, and cecal fermentation over single ingredient. Consistently, this combination increased total short-chain fatty acids production, particularly acetate, improved gut tight-junction organization and reduced myeloperoxidase accumulation. Shotgun metagenomic sequencing indicated that this combination reshaped the gut microbiota, notably enriching Anaeromassilibacillus sp., which associated with improved clinical feature. These findings highlight the potential of fungal-derived {beta}-glucan and probiotic combinations as functional food ingredients for mitigating intestinal inflammation and supporting barrier repair.

Background: Prediabetes is an intermediate state of increased glucose concentrations above the normal range but below the diagnostic threshold for type 2 diabetes (T2D). Adults with overweight and prediabetes are at increased risk of progression to T2D and metabolic imbalance. Feiolix -a whole-fruit feijoa powder rich in abscisic acid, polyphenols and dietary fibre which may support metabolic health with complementary pathways. Objective: To evaluate the effects of low and high-dose Feiolix compared with placebo, on HbA1C, glucagon like peptide (GLP-1) and other metabolic parameters. Methods: Adults aged 21-65 years with prediabetes i.e. glycosylated hemoglobulin (HbA1c) 5.7-6.4% and overweight with body mass index (BMI)- 25.0-29.9 kg/m2 were randomised to receive Feiolix-LD (300 mg/day) or Feiolix-HD (1,150 mg/day), or placebo (maltodextrin) for 12 weeks. Anthropometric measurements were obtained monthly, and fasting blood samples were collected at baseline and 12 weeks post intervention. Postprandial blood samples were collected at baseline and 12 weeks. Outcomes included HbA1c, GLP-1, fasting plasma glucose, insulin resistance lipid markers, body weight and BMI. Analyses were conducted using ANCOVA model with baseline values as covariates. Result: Of 41 individuals screened, 39 were randomised (13 per group); one participant withdrew from the Feiolix-LD. At week 12, HbA1c decreased by 0.5% in the Feiolix-HD and by 0.4% in the Feiolix-LD, with both reductions significantly greater than placebo (p < 0.05); Compared with placebo, Feiolix supplementation significantly lowered fasting glucose, insulin, HOMA-IR, total cholesterol and LDL-cholesterol. The Feiolix-HD significantly attenuated the increase in triglycerides observed in the placebo group and was associated with significant reductions in body weight and BMI versus placebo by month 2. However, no change in GLP-1 was observed between the treatments. No treatment-related adverse events were reported. Conclusion: This study supports the potential of Feiolix supplementation to improve metabolic health, including glycemic and lipid outcomes, with additional benefits for body weight and BMI at the higher dose. This study supports the potential of Feiolix supplementation to improve metabolic health, including glycemic and lipid outcomes, with additional benefits for body weight and BMI at the higher dose

Background & Aims Personalized nutrition has the potential to improve nutritional intake and clinical outcomes in hospitalized patients by adapting to individual dietary needs and preferences. Despite its promise, no personalized nutrition product (PNP) for hospitalized patients is currently available. This study aimed to assess the acceptability of PNPs among hospitalized patients and hospital staff. Methods A prospective, ten-days, proof-of-concept study was conducted among in-hospital adult patients ([&ge;]18y). Study patients received a freshly 3D printed PNP, customized to match individual taste preferences and health profiles. Patients could reorder the product daily if they remained hospitalized >24h after consumption. Consumption was assessed by weighing the PNP before and after consumption. Acceptability, sensory feedback, and overall experience were evaluated using questionnaires. Hospital staff also evaluated sensory feedback and acceptability. Factors influencing consumption and liking among patients were evaluated by testing within-group differences. Correlations between consumption rates, liking and patient characteristics were assessed. Results Eighty-seven patients received 137 PNPs during the study period. The average overall liking score was 5.7 ({+/-}2.1) on a 1-9 scale, and median consumption rate was 51% [Interquartile Range 19-100]. Consumption rates positively correlated with overall liking (R2 = 0.50, p < 0.001). Patients' willingness to reorder was 3.5 ({+/-}2.3) on a 1-7 scale, and reorder rate 63%. The perceived value of a PNP service in a hospital setting (scale 1-7) was scored 5.3 ({+/-}1.7) by patients and 4.5 ({+/-}1.1) by staff. Conclusions PNPs were evaluated as moderately positive in terms of acceptability and value by both patients and staff. However, further optimization of the product is needed before evaluating clinical efficacy.

Nisin is a promising antimicrobial peptide widely used in food preservation due to its efficacy against Gram-positive spoilage and pathogenic bacteria. Although Nisin is increasingly applied in the food sector, the biopeptide suffers from instability within food matrixes and can rapidly lose its antimicrobial potential following interaction with food biomolecules. Thus, it is necessary to investigate approaches that can be employed to extend the stability and activity of Nisin. Hence, the aim of this study was to develop and characterise a chitosan-alginate polyelectrolyte microencapsulation system capable of enhancing Nisin stability while retaining antimicrobial activity. The microencapsulation of Nisin was achieved by pre-gelation of alginate using calcium chloride and subsequent direct electrostatic interaction between cationic Nisin and chitosan with pre-gelled anionic alginate at pH 5.0. Following microcapsule formation, physicochemical and structural characterisation was performed using Zeta potential determination and measurement of the polydispersity index (PDI) via dynamic light scattering. SEM micrographs were used to confirm morphology, while Fourier-transform infrared (FTIR) spectroscopy and high-performance liquid chromatography (HPLC) were utilised to assess chemical integrity and functional group preservation of encapsulated Nisin. Following this, stable microcapsules with diameters ranging from 150-200 nm and smooth surface morphology were obtained. Microcapsule formation was strongly influenced by formulation parameters, particularly pH, calcium ion concentration, and chitosan content, with deviations from optimal acidic conditions (< pH 5.0) resulting in aggregation, increased polydispersity, and reduced encapsulation efficiency. The microcapsules were monodispersed (PDI {approx} 0.30) and electrostatically stable, exhibiting a Zeta potential of approximately +36 mV. These microcapsules remained stable over a prolonged storage period of 21 days under refrigerated conditions while retaining antimicrobial activity against Bacillus cereus. Encapsulation efficiency reached approximately 65%, confirming effective retention of Nisin within the polymer matrix. Overall, the findings demonstrate that chitosan-alginate ionic gelation is a non-denaturing and effective encapsulation strategy for extending the functional stability of Nisin. These microcapsules show strong potential as natural antimicrobial delivery systems for food and beverage applications, particularly in acidic food matrices, with implications for improved food safety and shelf-life extension.

Introduction: Study to evaluate the pharmacokinetics (PK), subject satisfaction, and gastrointestinal (GI) tolerability of single and multiple doses of a novel oral iron formulation based on iron enriched microalgae Spirulina platensis supplemented with inactivated yeast. Methods: Twelve healthy women with baseline serum iron 4-12 mol/L (22.3-67.0 g/dL) were admitted to the study site for 3 days in this single centre, multiple-dose, open-label study. Subjects received an oral dose of Monurelle(R) Ferro Naturale (MFN) on Days 1, 2, and 3. Blood for PK analysis was collected pre-dose (0) and at 1, 2, 3, 4, 5, 8 hours after dosing on Days 1 and 3, with assessment of subject satisfaction and GI tolerability performed 3 hours after each dose. Results: Both single and multiple dosing with MFN led to a defined peak level (Cmax) and increased systemic exposure (AUC0-t), with serum iron rising to almost double the circulating amount at 5 h post-dose (tmax). Mean ({+/-}SD) percent change from baseline at 5 h post-dose was 94.7% ({+/-}56.1) on Day 1 and 72.0% ({+/-}70.7) on Day 3. The majority of subjects reported being satisfied with the product and would use it again in the future. MFN was very well tolerated, with only one adverse event (AE) reported (episode of vomiting after the first dose, that lasted for one minute and did not recur after the two further doses of MFN). No other subject experienced any GI side effects. Discussion: This first report of oral Monurelle(R) Ferro Naturale in humans shows this novel formulation to provide a favourable PK profile with respect to serum iron concentrations, with excellent tolerability and high subject satisfaction.

Previous research has shown that mammalian fatty acids (FAs) can influence antibiotic tolerance of Staphylococcus aureus, yet many of these studies overlook the sources of these FAs, which are primarily esterified into glycero- and phospholipids, and the impact S. aureus lipase activity has on host lipids. Here we attempt to gain insight into the complex interplay between the S. aureus lipidome and its environment using culture media supplemented tissue-specific phospholipid mixtures. Phospholipid profiles of heart, liver, and brain-derived lipids revealed distinct distributions of headgroup and fatty acyl tail structures within the phospholipids. Following the growth of S. aureus in lipid-enriched broth, PG species containing mono- and poly-unsaturated acyl tails were detected with abundances that correlated strongly with the FA profile of the tissue extract. We found that S. aureus cultured with liver-derived lipid extract, which yielded the most unsaturated PGs, promoted growth in high concentrations of the membrane-targeting antimicrobial daptomycin. To explore the influence of lipase activity on the extracellular lipids, comparative analysis of fresh versus spent media revealed that the lipase-mediated degradation of complex phospholipid mixtures was influenced by both head group structure and acyl tail linkage. Concurrently, the spent media contained elevated levels of mono- and polyunsaturated lysophospholipids that were predominantly of the 2-acyl form rather than the 1-acyl form observed in the fresh media. Together, these results demonstrate the extent to which the lipase activity of S. aureus remodels both its own lipidome as well as the structures of the phospholipids in the surrounding environment. IMPORTANCES. aureus releases a secreted glycerol ester hydrolase, Geh, into the extracellular environment, which enables the bacterium to generate free FA from glycerolipids, phospholipids, and cholesterol esters that are present in surrounding tissue of an infection. The liberated FAs can be incorporated into the phospholipids of S. aureus, thereby altering its membrane physiology with mono- and poly-unsaturated FAs it cannot otherwise synthesize. Simultaneously, the action of Geh on lipids in the host environment leads to higher levels of bioactive lysophospholipids that participate in mammalian signaling pathways. This work reveals the preferences of S. aureus Geh across phospholipids with different head group and acyl tail structures found within tissue-derived lipid extracts, as well as the fate of the liberated FAs within the staphylococcal membrane lipids. The impacts of these processes on both the host and bacterium have implications for the immune response to and antibiotic treatment of S. aureus infections.

Gut dysbiosis poses a significant public health concern, and malnutrition plays a central role in its development. In our recent clinical trial, we explored the effects of regulated walnut intake on the levels of 11 fecal metabolites to determine whether (I) walnut consumption promotes changes in metabolite profiles consistent with reduced gut dysbiosis, and (II) whether these changes are associated with the levels of urolithin A, a bioactive metabolite produced by colonic microbes following walnut consumption. DNA was extracted from fecal samples using a DNeasy 96 PowerSoil Pro QIAcube HT for 16S rRNA sequencing, and fecal short-chain fatty acids (SCFAs) and p-cresol were extracted fecal specimens by liquid chromatography/mass spectrometry (LC/MS-MS). At baseline, obese individuals (BMI > 30) had lower alpha diversity, higher p-cresol and valeric acid levels, and a higher trending Firmicutes to Bacteroidetes (F/B) ratio compared to non-obese (BMI < 30) participants, indicating a level of inherent gut dysbiosis. Walnut intake was associated with an overall 2.8% increase in alpha diversity, a 17.6% reduction in estimated marginal mean F/B ratio, and an overall reduction in three luminal proteolytic metabolites associated with gut dysbiosis: isobutyric acid, valeric acid, and isovaleric acid. Interestingly, the effects on the gut metabolome were more pronounced in obese individuals, with significant decreases observed in the same three SCFAs, as well as p-cresol. Given the known associations between these luminal proteolytic metabolites and gut dysbiosis, these results suggest that walnut intake may help to attenuate overall gut dysbiosis, particularly in obese populations.

Consumption of ultra-processed food (UPF) is increasing and is causally linked with noncommunicable disease onset, however specific compounds within UPFs that influence disease risk remain poorly defined. The acronym 'SCars', for Stable Carbonyl Adducts, is introduced herein as an umbrella term encompassing advanced glycation end-products (AGEs), advanced lipoxidation end-products (ALEs), and other carbonyl-derived modifications. These molecules arise as products of spontaneous carbonyl chemistry, a reaction accelerated by industrial food processing. Dietary SCars have been associated with metabolic dysfunction; however, their impact on microbiome composition during windows of developmental vulnerability has not been well studied. Puberty in mice represents a period of developmental plasticity during which the gut microbiome is susceptible to dietary exposures. We tested whether transient exposure to high levels of dietary SCars during puberty remodels the developing gut microbiome. Using metagenomic sequencing of fecal samples from mice exposed to high-SCars or control diets, we found that high-SCars exposure profoundly impacted microbial community structure and metabolic potential. Dietary SCars reduced microbial diversity and depleted short-chain fattyacid producing taxa while enriching pathways related to membrane remodeling, branched-chain amino acid biosynthesis, and nucleotide anabolism. These data identify SCars as features of UPFs capable of reprogramming the developing gut microbiota.

Growing evidence links ultra-processed food intake according to the Nova classification with poor health, yet no comprehensive database of additives and food substances of non-culinary use that are markers of ultra-processing exists for the UK. Building the database involved seven steps: (1) obtaining all approved additives in the UK, (2) retrieving all of their technical functions, (3) coding each technical function as cosmetic (marker of ultra-processing) or non-cosmetic (not a marker of ultra-processing), (4) adding 'flavour extracts', 'natural flavourings', and 'flavourings', (5) excluding additives as ultra-processing markers if they served fortification purposes, or were a herb/spice, and (6) adding food substances of non-culinary use defined as markers of ultra-processing in the Nova classification. Lastly, (7) two UPF definitions were constructed based on the ambiguity/overlap of additive technical functions from previous approaches. The database is freely accessible for researchers to improve consistency in using the Nova classification in the UK.

Lactoferrin is a multifunctional iron-binding glycoprotein that supports intestinal barrier function, immune regulation, and a favorable gut microbial environment. However, the contribution of gut microbial biotransformation to its gastrointestinal activity remains poorly understood. We investigated whether effera(R), a precision fermentation-derived recombinant human lactoferrin, supports intestinal barrier function through microbiome-mediated mechanisms. effera(R) underwent simulated upper gastrointestinal digestion followed by ex vivo colonic fermentation using the validated SIFR(R) technology pipeline, which employs bioreactors that are inoculated with fecal microbiota from six healthy adult donors. Microbial activity was evaluated by measuring short-chain fatty acid (SCFA) production, bacterial cell density, and microbiome composition. effera(R) produced dose-dependent increases in the production of SCFAs and bacterial cell density demonstrating enhanced microbial metabolic activity. These metabolic changes were accompanied by shifts in key microbial groups within Bacillota_A and Bacteroidota. Intact effera(R) and cell-free post-colonic fermentation-derived products were evaluated in a Caco-2/THP-1 epithelial-immune co-culture model under basal and lipopolysaccharide-challenged conditions. Whereas intact protein did not significantly improve epithelial barrier integrity, effera(R)s post-colonic fermentation-derived products significantly enhanced transepithelial electrical resistance (TEER) under basal conditions and produced an even stronger barrier-protective response following LPS challenge. Across matched doses, effera(R) consistently generated greater TEER responses than bovine lactoferrin. Improved barrier function was accompanied by increased expression of tight-junction-associated targets ZO-1 and occludin and reduced secretion of CXCL-10 and IL-8. Together, these findings demonstrate that microbial biotransformation enhances the biological activity of effera(R), linking increased microbial metabolism with improved epithelial barrier integrity and modulation of inflammatory signaling. This integrated study provides a strong mechanistic foundation for the use of human lactoferrin in adult gut-health applications and offers valuable guidance for future adult clinical studies and infant-relevant investigations.

Biopreservatives including nisin and its derivatives are becoming more desirable in the food processing industry because of the growing demand for naturally preserved and minimally processed foods free from artificial preservatives. However, ensuring microbiological safety while meeting these consumer preferences remains a major challenge. This has necessitated the continuous investigation of potential new antimicrobial agents produced by naturally occurring microorganisms. Hence, this study explored the synthesis, characterisation, and optimisation of a bacteriocinogenic lactic acid bacterium and its antimicrobial product, possibly novel bacteriocin (Nisin 2A) from Lactococcus lactis isolated from commercial brined cheese. The isolation was achieved by screening for wild-type bacteriocin-producing lactic acid bacteria from dairy products using MRS media. Screening was performed using antagonism assays, yielding five producer organisms. Of these, the isolate whose metabolites exhibited the most potent antimicrobial activity was identified as Lactococcus lactis, which synthesised an active antimicrobial peptide designated as Nisin 2A, with a molecular mass of approximately 3.3 kDa as determined by UHPLC-MS and SDS-PAGE. Production of Nisin 2A was scaled up through fed-batch fermentation of Lactococcus lactis in modified MRS broth following process optimisation using a Plackett-Burman experimental design and purified by ammonium sulphate precipitation and solid-phase extraction (SPE). Furthermore, the antimicrobial potential of the bacteriocin was evaluated by the agar well diffusion assay and quantified using the tube dilution method. The purified peptide demonstrated broad-spectrum antimicrobial activity, particularly against the test Gram-positive bacteria Bacillus cereus and retained its bioactivity across a wide pH range (3-9) and high thermal conditions (up to 100 {degrees}C). Furthermore, it had high sensitivity to proteolytic enzymes (Proteinase K and Trypsin). Notably, the peptide was thermostable and retained up to 90% of its initial activity after thermal treatment and maintained consistent inhibitory performance after extended storage. These findings highlight the potential application of Nisin 2A as a natural biopreservative in food systems.

AO_SCPLOWBSTRACTC_SCPLOWO_ST_ABSIntroductionC_ST_ABSPolyunsaturated fatty acid (PUFA) status is an important marker of metabolic health across the lifespan, and omega-3 (n-3) PUFA were shown to have anti-inflammatory and cardioprotective effects. The omega-3 index, a composite of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), is widely used to assess n-3 PUFA supplementation status, and product-to-precursor fatty acid ratios are used as proxy indices of desaturation and elongation. Previous data indicate a higher proportion of n-3 PUFA with age. Whether the same pattern can be found in an NHANES cohort, and whether long-chain n-3 status relates to phenotypic age (PhenoAge), a composite measure of biological age, has not been established. MethodsWe analyzed erythrocyte fatty acid profiles measured by electron capture negative-ion mass spectrometry in erythrocytes of participants of the National Health and Nutrition Examination Survey (NHANES) 2021-2023. Adults aged 20-79 years with a complete 21-fatty-acid panel were included (n = 4,863) and stratified into five age groups (20-34, 35-44, 45-54, 55-64, 65-79 years). In the fasting subsample with a measured fasting glucose (n = 2,822) we additionally computed PhenoAge according to Levine and assessed PhenoAge acceleration (PhenoAge regressed on chronological age) in relation to EPA and DHA. ResultsPercentage of summarized PUFA and n-6 PUFA decreased with age while percentage of n-3 PUFA and EPA + DHA increased. In contrast, total saturated fatty acids did not differ across age groups. EPA and DHA rose with age, whereas linoleic acid (LA), dihomo-{gamma}-linolenic acid (DGLA) and -linolenic acid (ALA) declined. The delta-5-desaturase (D5D) index (AA/DGLA) and the AA/LA ratio were positively associated with age, while elongation-related proxies ELOVL2 (DTA/AA, DPA/EPA) and ELOVL5 (DGLA/GLA) were inversely associated. Higher EPA + DHA was associated with lower PhenoAge acceleration with the most pronounced effect of - 4,7 years in the group with highest ([&ge;]8%) EPA+DHA content. DiscussionAge-related shifts in erythrocyte PUFA composition and in desaturase and elongase proxy indices reported previously in a clinical cohort from Germany are reproduced in a large US population. In addition, long-chain n-3 status was inversely associated with PhenoAge acceleration.

BackgroundEmerging evidence suggests that PFAS can cross blood-brain barrier and lead to neurotoxicity. Recent evidence also suggest that PFAS can bioaccumulate in gut microbiota resident in the gut. However, how gut microbes influence PFAS-driven reorganization of metabolic homeostasis in the brain is poorly understood. MethodsTo address this gap, we investigated how gut microbiota influences brain metabolomic and lipidomic responses to PFAS exposure. Specific pathogen-free (SPF) and germ-free (GF) mice were fed an obesogenic diet for 8 weeks to promote metabolic disturbance. After 1 week of acclimation, half received control water and half received water containing a PFAS mixture (PFHxS, GenX, PFOA, PFOS, and FTOH mixture). Plasma and brain samples (cortex, subcortex, cerebellum, olfactory bulb, and brainstem) were collected after 8 weeks. Untargeted analyses were performed for lipidomic, metabolomic and PFAS using high resolution liquid chromatography tandem mass spectrometry (LC-MS/MS). Data was processed using MassCube with open-sources libraries. ResultsPFHxS, GenX, PFOA, PFOS, PFDA, and PFDS were detected in plasma. PFHxS, PFOA, PFOS, and PFDS were detected across all five brain regions, with PFOS as the predominant brain-enriched species. Pathway analysis identified nicotinate and nicotinamide metabolism as the most consistently PFAS-altered pathway in both SPF and GF mice. PFAS exposure induced region-specific metabolic remodeling, with gut microbiota differentially modulating responses in the cortex, cerebellum, and brainstem, whereas the olfactory bulb showed a largely microbiota-independent response. In addition to local effects within individual brain regions, plasma-brain analysis suggested systemic metabolic responses across tissues, with association strength varying by brain region and microbiome status. Gut microbiota also shaped PFAS-induced lipid dysregulation in the brain, and methylnicotinamide and delta-valerobetaine were among the most responsive metabolites. ConclusionThis study is the first to demonstrate that resident microbiota impact PFAS-associated metabolic remodeling across the gut-plasma-brain axis. HighlightsO_LIPFAS-induced metabolic remodeling in the brain is modified by gut microbiota. C_LIO_LIPFAS exposure alters nicotinate and nicotinamide metabolism throughout the brain. C_LIO_LIPFAS-induced brain metabolic responses are region specific and microbiota dependent. C_LIO_LIPlasma-brain analysis suggests potential systemic metabolic disruption by PFAS. C_LI Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=104 SRC="FIGDIR/small/743341v1_ufig1.gif" ALT="Figure 1"> View larger version (38K): org.highwire.dtl.DTLVardef@15301deorg.highwire.dtl.DTLVardef@9fac0aorg.highwire.dtl.DTLVardef@d7f0f4org.highwire.dtl.DTLVardef@10c29c2_HPS_FORMAT_FIGEXP M_FIG C_FIG

Abstract: Background: Dietary intervention can modulate the composition of gut microbial community. However, the consistency and magnitude of these changes may differ between individuals. Emerging evidence indicates that such variation is influenced by the baseline microbial composition and host characteristics, while variation in diet, lifestyle, and environmental exposures can further contribute to this heterogeneity. Shared-living settings, in which participants co-reside at a common site and consume a standardized diet, provide an opportunity to minimize variability arising from these external factors and to investigate the factors of microbiome composition and response to diet. In this study, we characterized the determinants of baseline gut microbial composition and evaluated the response to a short-term gut-friendly dietary intervention in a multi-ethnic cohort living under shared-living conditions. Methods. Forty-three adults from North India, South India and international backgrounds co-lived in a "pop-up village" for 15 days in a single-arm design. Participants followed a gut-friendly dietary regimen. Participants were stratified by body mass index (BMI) as overweight/obese (OW, [&ge;] 25) or non-overweight (Non-OW, < 25). Stool samples collected at baseline and at the end of the dietary intervention (43 participants, 86 samples) were profiled by Oxford Nanopore shotgun metagenomics. Samples were evaluated for taxonomic composition, alpha diversity (per-participant linear mixed-effects models), differential abundance (consensus across three tools: MaAsLin2, LinDA, ALDEx2), and assembly-based functional gene content for six important metabolic pathways. Results. At baseline, geographic region (p = 0.046) and BMI (p = 0.024) were the main factors to structure the community composition, whereas habitual diet type was not (p = 0.792). The intervention produced a small but highly reproducible shift, significant across all five distance metrics (p [&le;] 0.003). The response was stronger among overweight (OW) participants, who showed a significant shift (p = 0.001), whereas non-overweight (Non-OW) participants did not (p = 0.181). The Timepoint with BMI interaction was significant for Bray-Curtis (p = 0.021) and Aitchison (p = 0.028) distances. Whole-cohort differential-abundance analysis identified depletion of the oral-associated species Streptococcus parasanguinis (q = 0.010) and Gemella sanguinis (q = 0.021), together with differential abundance of six genera, including decreased Actinomyces (q = 0.004) and increased Faecalibacterium (q = 0.016). Functional community composition of six major metabolic pathways remained largely stable at the whole-cohort level. Conclusion. A short, shared gut-friendly diet elicited a modest but coordinated shift in gut microbiome composition, characterized by depletion of several oral-associated pathobiont taxa and enrichment of fibre- fermenting taxa, with the community-level compositional shift observed predominantly among overweight participants. Keywords: Gut Microbiome, Dietary intervention, Shotgun Metagenomics, Pop-Up Village Study.

Kakadu plum (Terminalia ferdinandiana), an Australian native fruit, is among the richest known dietary sources of vitamin C and hydrolysable tannins, yet its capacity to protect the intestinal epithelium against oxidative stress remains largely unexplored. This study optimised the extraction of bioactive compounds from freeze-dried Kakadu plum powder and evaluated their antioxidant activity using both chemical and cellular antioxidant in vitro assay. Phenolic compounds were extracted using three solvents (water, 80% ethanol, and 80% methanol) combined with shaking, ultrasound, or microwave assistance. Solvent, rather than processing technique, was the dominant determinant of antioxidant capacity: ethanol and methanol maximised total phenolic content, total flavonoid content, and DPPH radical-scavenging activity, whereas water extracts showed the highest ferric-reducing antioxidant power. Twenty-four phenolic compounds identified by HPLC-ESI-QTOF-MS/MS were mapped by network pharmacology to nine core oxidative-stress targets, and cross-species molecular docking predicted conserved binding of key phenolics to canine orthologs of PTGS2 and MMP2. In an H2O2-induced oxidative-stress in vitro cell model using canine small intestinal epithelial cells, both water (less than 25 ug/mL) and ethanol (less than 250 ug/mL) extracts significantly suppressed intracellular reactive oxygen species (ROS) in a dose-dependent manner, with the ethanol extract effective across a wider concentration range. This work demonstrated that Kakadu plum extract could be a promising natural, multi-target antioxidant ingredient for canine intestinal health, and provided a reference for future in vivo research.

Fiber supplements are the first line of treatment for all patients with benign anorectal disease. In addition, fiber has been found to support gut microbiota and reduce inflammation; however, their integrated effect on the colonic mucosal environment is incompletely defined. We evaluated how mixed-fiber formulations may collectively shape the rectal mucosal environment through coordinated effects on microbiota composition, short-chain fatty acid (SCFA) handling, and host transcription. We evaluated multi-compartment responses to a 28-day mixed-fiber intervention in 37 participants with benign anorectal disease (~15 g/day; 7.5g psyllium husk, 8g wheat dextrin) and assessed the microbiota (16S rRNA), the metabolome (GC-MS and LC-MS), and transcriptome (subset n=10) before (PRE) and after (POST) supplementation. Changes in KEGG pathways were tested with gene set enrichment analysis (GSEA). Associations were tested by Spearman correlation, and microbiota-based prediction of SCFA responses by random-forest regression. Microbiota diversity was stable after the fiber intervention, however, 10 mucosal ASVs were differentially abundant including Parabacteroides and Lachnospiraceae NK4A136 group while SCFAs, propionate and isobutyrate, both decreased in rectal tissue and butyrate decreased in serum. To understand these shifts in the metabolome, we proceeded with the transcriptome where we identified 130 differentially expressed genes and enrichment of metabolic and SCFA-related pathways. Random forest regression captured a modest signal for serum butyrate. Taken together, short-term mixed-fiber supplementation produced selective rectal mucosal taxonomic shifts and enriched mucosal metabolic and SCFA-metabolism program despite reduced SCFA pools, suggesting that even brief over-the-counter fiber supplementation may reshape the mucosal metabolic environment.