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Transaminases (EC 2.6.1.X, TAs) are important biocatalysts in the synthesis of chiral amines, and have significant value in the field of medicine. However, TAs suffer from low enzyme activity and poor catalytic efficiency in the synthesis of chiral amines or non-natural amino acids, which hinders their industrial applications. In this study, a novel TA derived from Paracoccus pantotrophus (ppTA) that was investigated in our previous study was employed with a semi-rational design strategy to improve its enzyme activity to 2-ketobutyrate. By using homology modeling and molecular docking, four surrounding sites in the substrate-binding S pocket were selected as potential mutational sites. Through alanine scanning and saturation mutagenesis, the optimal mutant V153A with significantly improved enzyme activity was finally obtained, which was 578 % higher than that of the wild-type ppTA (WT). Furthermore, the mutant enzyme ppTA-V153A also exhibited slightly improved temperature and pH stability compared to WT. Subsequently, the mutant was used to convert 2-ketobutyrate for the preparation of L-2-aminobutyric acid (L-ABA). The mutant can tolerate 300 mM 2-ketobutyrate with a conversion rate of 74 %, which lays a solid foundation for the preparation of chiral amines., Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Objective: Chronic exposure to cold temperature is known to elevate blood pressure, leading to a condition known as cold-induced hypertension (CIH). Our previous research suggested correlations between alterations in gut microbiota, decrease in butyrate level, and the onset and progression of CIH. However, the role of butyrate in CIH and the underlying mechanisms need further investigation., Methods: We exposed Specific Pathogen Free (SPF) rats to continuous cold temperature (4 ± 1 °C) for 6 weeks to establish a CIH rat model. Rats were divided into different groups by dose and duration, and the rats under cold were administered with butyrate (0.5 or 1 g/kg/day) daily. We assessed hypertension-associated phenotypes, pathological morphological changes, and endocrine-related phenotypes of brown adipose tissue (BAT). The effects of butyrate on gut microbiota and intestinal content metabolism were evaluated by 16s RNA sequencing and non-targeted metabolomics, respectively., Results: The systolic blood pressure (SBP) of rats exposed to cold after supplemented with butyrate were significantly lower than that of the Cold group. Butyrate may increase the species, abundance, and diversity of gut microbiota in rats. Specifically, butyrate intervention enriched beneficial bacterial genera, such as Lactobacillaceae, and decreased the levels of harmful bacteria genera, such as Actinobacteriota and Erysipeiotrichaceae. Cold exposure significantly increased BAT cells and the number of mitochondria. After butyrate supplementation, the levels of peroxisome proliferator-activated receptor gamma coactivator 1a and fibroblast growth factor 21 in BAT were significantly elevated (P < 0.05), and the volume and number of lipid droplets increased. The levels of ANG II and high-density lipoprotein were elevated in the Cold group but decreased after butyrate supplementation., Conclusion: Butyrate may attenuate blood pressure in CIH by promoting the growth of beneficial bacteria and the secretion of beneficial derived factors produced by BAT, thus alleviating the elevation of blood pressure induced by cold. This study demonstrates the anti-hypertensive effects of butyrate and its potential therapeutic mechanisms, offering novel insights to the prevention and treatment of CIH in populations living or working in cold environments., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest., (Copyright © 2024. Published by Elsevier B.V.)

Background and Aims: The PROMINENT trial, a cardiovascular outcome trial of the triglyceride- and remnant cholesterol-lowering agent pemafibrate, has shown neutral results despite reduction in plasma triglycerides and remnant cholesterol. We tested the hypothesis that absolute mass changes in remnant cholesterol, LDL cholesterol, and apolipoprotein B explain the results of the PROMINENT trial., Methods: Among 108,431 individuals from the Copenhagen General Population Study (CGPS), those who met the key inclusion criteria of the PROMINENT trial were analyzed to mimic the trial design. Endpoint atherosclerotic cardiovascular disease (ASCVD) was cardiovascular death, myocardial infarction, ischemic stroke, and coronary revascularization as defined in PROMINENT., Results: In the PROMINENT trial, treatment with pemafibrate resulted in -7 mg/dL (-0.18 mmol/L; -18 %) change in remnant cholesterol, +10 mg/dL (+0.26 mmol/L; +12 %) LDL cholesterol, and +5 mg/dL (+0.05 g/L; +5 %) apolipoprotein B. In the CGPS mimicking PROMINENT, the estimated hazard ratios for ASCVD were 0.97 (95 % confidence interval: 0.94-0.99) for a -7 mg/dL (-0.18 mmol/L) change in remnant cholesterol, 1.04 (1.01-1.07) for a +10 mg/dL (+0.26 mmol/L) change in LDL cholesterol, and 1.02 (1.01-1.03) for a +5 mg/dL (+0.05 g/L) change in apolipoprotein B. When combining absolute changes in remnant cholesterol, LDL cholesterol, and apolipoprotein B, the estimated hazard ratio for ASCVD was 1.05 (0.96-1.14) in the CGPS mimicking PROMINENT compared to 1.03 (0.91-1.15) in the PROMINENT trial., Conclusions: Absolute mass changes in remnant cholesterol, LDL cholesterol, and apolipoprotein B can explain results of the PROMINENT trial. The 3 mg/dL (0.08 mmol/L) higher total atherogenic cholesterol together with 5 mg/dL (0.05 g/L) higher apolipoprotein B seem to explain the trend toward more ASCVD in the pemafibrate arm., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: BGN reports consultancies or talks sponsored by AstraZeneca, Sanofi, Regeneron, Akcea, Amgen, Amarin, Kowa, Denka, Novartis, Novo Nordisk, Esperion, Silence Therapeutics, Abbott, Mankind, Lilly, and Arrowhead. TD reports a consultancy by Novo Nordisk. AL reports a talk sponsored by Amarin and a consultancy by Novartis., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

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., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

IlvA1, a pyridoxal phosphate-dependent (PLP) enzyme, catalyzes the deamination of l-threonine and l-serine to yield 2-ketobutyric acid or pyruvate. To gain insights into the function of IlvA1, we determined its crystal structure from Pseudomonas aeruginosa to 2.3 Å. Density for a 2-ketobutyric acid product was identified in the active site and a putative allosteric site. Activity and substrate binding assays confirmed that IlvA1 utilizes l-threonine, l-serine, and L-allo-threonine as substrates. The enzymatic activity is regulated by the end products l-isoleucine and l-valine. Additionally, the efficiency of d-cycloserine and l-cycloserine inhibitors on IlvA1 enzymatic activity was examined. Notably, site-directed mutagenesis confirmed the active site residues and revealed that Gln165 enhances the enzyme activity, emphasizing its role in substrate access. This work provides crucial insights into the structure and mechanism of IlvA1 and serves as a starting point for further functional and mechanistic studies of the threonine deaminase in P. aeruginosa., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Background: Predicting response to exclusive enteral nutrition (EEN) in active Crohn's disease (CD) could lead to therapy personalization and pretreatment optimization., Objectives: This study aimed to explore the ability of pretreatment parameters to predict fecal calprotectin (FCal) levels at EEN completion in a prospective study in children with CD., Methods: In children with active CD, clinical parameters, dietary intake, cytokines, inflammation-related blood proteomics, and diet-related metabolites, metabolomics and microbiota in feces, were measured before initiation of 8 wk of EEN. Prediction of FCal levels at EEN completion was performed using machine learning. Data are presented with medians (IQR)., Results: Of 37 patients recruited, 15 responded (FCal < 250 μg/g) to EEN (responders) and 22 did not (nonresponders). Clinical and immunological parameters were not associated with response to EEN. Responders had lesser (μmol/g) butyrate [responders: 13.2 (8.63-18.4) compared with nonresponders: 22.3 (12.0-32.0); P = 0.03], acetate [responders: 49.9 (46.4-68.4) compared with nonresponders: 70.4 (57.0-95.5); P = 0.027], phenylacetate [responders: 0.175 (0.013-0.611) compared with nonresponders: 0.943 (0.438-1.35); P = 0.021], and a higher microbiota richness [315 (269-347) compared with nonresponders: 243 (205-297); P = 0.015] in feces than nonresponders. Responders consumed (portions/1000 kcal/d) more confectionery products [responders: 0.55 (0.38-0.72) compared with nonresponders: 0.19 (0.01-0.38); P = 0.045]. A multicomponent model using fecal parameters, dietary data, and clinical and immunological parameters predicted response to EEN with 78% accuracy (sensitivity: 80%; specificity: 77%; positive predictive value: 71%; negative predictive value: 85%). Higher taxon abundance from Ruminococcaceae, Lachnospiraceae, and Bacteroides and phenylacetate, butyrate, and acetate were the most influential variables in predicting lack of response to EEN., Conclusions: We identify microbial signals and diet-related metabolites in feces, which could comprise targets for pretreatment optimization and personalized nutritional therapy in pediatric CD., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Parkinson's disease (PD) is intricately linked to abnormal gut microbiota, yet the specific microbiota influencing clinical outcomes remain poorly understood. Our study identified a deficiency in the microbiota genus Blautia and a reduction in fecal short-chain fatty acid (SCFA) butyrate level in PD patients compared to healthy controls. The abundance of Blautia correlated with the clinical severity of PD. Supplementation with butyrate-producing bacterium B. producta demonstrated neuroprotective effects, attenuating neuroinflammation and dopaminergic neuronal death in mice, consequently ameliorating motor dysfunction. A pivotal inflammatory signaling pathway, the RAS-related pathway, modulated by butyrate, emerged as a key mechanism inhibiting microglial activation in PD. The change of RAS-NF-κB pathway in PD patients was observed. Furthermore, B. producta-derived butyrate demonstrated the inhibition of microglial activation in PD through regulation of the RAS-NF-κB pathway. These findings elucidate the causal relationship between specific gut microbiota and PD, presenting a novel microbiota-based treatment perspective for PD., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Megasphaera hexanoica is a bacterial strain following the reverse β-oxidation pathway to synthesize caproate (CA) using lactate (LA) as an electron donor (ED) and acetate (AA) or butyrate (BA) as electron acceptors (EA). Differences in the type and concentration of EA lead to distinctions in product distribution and energy bifurcation of carbon fluxes in ED pathways, thereby affecting CA production. In this study, the effect of various ratios of AA, BA, and AA+BA as EA on carbon flux and CA specific titer during the carbon chain elongation in M. hexanoica was explored. The results indicated that the maximum levels of CA were 18.81 mM and 31.48 mM when the molar ratios of LA/AA and LA/BA were 10:1 and 3:1, respectively. Meanwhile, when AA and BA were used as combined EA (LA, AA, and BA molar amounts of 100, 23, and 77 mM), a maximum CA production of 39.45 mM was obtained. Further analysis revealed that the combined EA exhibited a CA production carbon flux of 49 % (4.3 % and 19.5 % higher compared to AA or BA, respectively) and a CA production specific titer of 45.24 mol (80.89 % and 58.51 % higher compared to AA or BA, respectively), indicating that the effective carbon utilization rate and CA production efficiency were greatly improved. Finally, a scaled-up experiment was conducted in a 1.2 L (working volume) automated bioreactor, implying high biomass (optical density at 600 nm or OD 600  = 1.809) and a slight decrease in CA production (28.45 mM). A decrease in H 2 production (4.11 g/m 3 ) and an increase in CO 2 production (0.632 g/m 3 ) demonstrated the appropriate metabolic adaptation of M. hexanoica to environmental changes such as stirring shear., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.)

l-threonate is the metabolite of vitamin C, while d-erythronate is the metabolite of N-acetyl-d-glucosamine, the nutritional supplement for joint health. They are widely distributed in the environment and human biofluids. Nevertheless, the catabolisms of l-threonate and d-erythronate are sparsely reported. Here we explored the functional diversity of an acid sugar kinase family (Pfam families PF07005-PF17042), and discovered a novel 2-oxo-tetronate kinase. The conserved genome neighborhood of the 2-oxo-tetronate kinase encodes members of class-II fructose-bisphosphate aldolase family (F&#95;bP&#95;aldolase, PF01116) and a dehydrogenase family (PF03446-PF14833). Instructed by this analysis, we experimentally verified that these enzymes are capable of degrading l-threonate into dihydroxyacetone phosphate (DHAP) in Arthrobacter sp. ZBG10, Clostridium scindens ATCC 35704, and Pseudonocardia dioxanivorans ATCC 55486. Meanwhile, a convergent catabolic pathway for d-erythronate was characterized in P. dioxanivorans ATCC 55486. Moreover, the phylogenetic distribution analysis indicates that the biological range of the identified l-threonate and d-erythronate catabolic pathways appears to extend mostly to members of the Actinomycetota, Cyanobacteriota, Bacillota, Pseudomonadota, and Bacteroidota phyla., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Hua Huang reports financial support was provided by National Natural Science Foundation of China. Xinshuai Zhang reports financial support was provided by National Natural Science Foundation of China. Xinshuai Zhang reports financial support was provided by Natural Science Foundation of Guangdong Province. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Background: Dietary fiber is an important health-promoting component of the diet, which is fermented by the gut microbes that produce metabolites beneficial for the host's health., Objectives: We studied the associations of habitual long-term fiber intake from infancy with gut microbiota composition in young adulthood by leveraging data from the Special Turku Coronary Risk Factor Intervention Project, an infancy-onset 20-y dietary counseling study., Methods: Fiber intake was assessed annually using food diaries from infancy ≤ age 20 y. At age 26 y, the first postintervention follow-up study was conducted including food diaries and fecal sample collection (N = 357). Cumulative dietary fiber intake was assessed as the area under the curve for energy-adjusted fiber intake throughout the study (age 0-26 y). Gut microbiota was profiled using 16S ribosomal ribonucleic acid amplicon sequencing. The primary outcomes were 1) α diversity expressed as the observed richness and Shannon index, 2) β diversity using Bray-Curtis dissimilarity scores, and 3) differential abundance of each microbial taxa with respect to the cumulative energy-adjusted dietary fiber intake., Results: Higher cumulative dietary fiber intake was associated with decreased Shannon index (β = -0.019 per unit change in cumulative fiber intake, P = 0.008). Overall microbial community composition was related to the amount of fiber consumed (permutational analysis of variation R 2 = 0.005, P = 0.024). The only genus that was increased with higher cumulative fiber intake was butyrate-producing Butyrivibrio (log 2 fold-change per unit change in cumulative fiber intake 0.40, adjusted P = 0.023), whereas some other known butyrate producers such as Faecalibacterium and Subdoligranulum were decreased with higher cumulative fiber intake., Conclusions: As early-life nutritional exposures may affect the lifetime microbiota composition and disease risk, this study adds novel information on the associations of long-term dietary fiber intake with the gut microbiota. This trial was registered at clinicaltrials.gov as NCT00223600., (Copyright © 2024 American Society for Nutrition. Published by Elsevier Inc. All rights reserved.)

Introduction: Chronic obstructive pulmonary disease (COPD) is associated with an intestinal leak and neuromuscular junction (NMJ) degradation, which contributes to physical compromise and accelerated age-related muscle loss, called sarcopenia. However, the relevant interventions partly remain ineffective. We investigated the effects of exogenous butyrate on sarcopenia and physical capacity with relevance to intestinal permeability and NMJ integrity in COPD patients., Methods: COPD patients were randomized into placebo (n = 67) and butyrate (n = 64) groups in a double-blind manner. The patients in the butyrate group received one 300 mg capsule a day for 12 weeks. We measured circulating markers of intestinal leak (zonulin), systemic bacterial load (LBP), and NMJ loss (CAF22), along with handgrip strength (HGS), and short physical performance battery (SPPB) at baseline and 12 weeks., Results: Butyrate supplementation improved HGS and gait speed in COPD patients. Among SPPB indices, butyrate improved the ability to maintain postural balance and walking and prevented a decline in the ability to rise from a chair. Butyrate also reduced the plasma levels of zonulin, LBP, and CAF22 levels in COPD patients (all p < 0.05). Regression analysis revealed significant associations of plasma zonulin and CAF22 with HGS, gait speed, and cumulative SPPB scores in butyrate group. These changes were associated with reduced markers of inflammation and muscle damage., Conclusion: Butyrate may provide a therapeutic approach to sarcopenia and physical dependency in COPD by repairing intestinal leak and NMJ loss., Competing Interests: Declaration of competing interest The authors declare that there is no conflict of interest., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Short-chain fatty acids (SCFAs), generated through microbial fermentation of dietary fibers and proteins in the gut, play a pivotal role in maintaining intestinal integrity, cellular function, and the immune response. SCFAs, including butyrate, acetate, and propionate, are absorbed in the colon or excreted through feces, contributing to essential physiological processes. Butyrate, a primary energy source for colonocytes, exhibits anti-inflammatory properties and regulates key pathways, such as nuclear factor-κB (NF-κB) inhibition. SCFAs' impact extends beyond the intestines, influencing the gut-brain axis, systemic circulation, and folate metabolism. A decline in colonic SCFAs has been linked to gastrointestinal diseases, emphasizing their clinical relevance, while their effects on immune checkpoints, such as ipilimumab, provide intriguing prospects for cancer therapy. This mini-review explores SCFAs' diverse roles, shedding light on their significance in health and potential implications for disease management. Understanding SCFAs' intricate mechanisms enhances our knowledge of their therapeutic potential and highlights their emerging importance in various physiological contexts., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Background: Wholegrain intake is associated with lower risk of cardiometabolic diseases in adults, potentially via changes in the gut microbiota. Although cardiometabolic prevention should start early, we lack evidence on the effects in children., Objectives: This study investigated the effects of wholegrain oats and rye intake on serum low-density lipoprotein (LDL) cholesterol and plasma insulin (coprimary outcomes), other cardiometabolic markers, body composition, gut microbiota composition and metabolites, and gastrointestinal symptoms in children with high body mass index (BMI)., Methods: In a randomized crossover trial, 55 healthy Danish 8- to 13-y-olds received wholegrain oats and rye ("WG") or refined grain ("RG") products ad libitum for 8 wk in random order. At 0, 8, and 16 wk, we measured anthropometry, body composition by dual-energy absorptiometry, and blood pressure. Fasting blood and fecal samples were collected for analysis of blood lipids, glucose homeostasis markers, gut microbiota, and short-chain fatty acids. Gut symptoms and stool characteristics were determined by questionnaires. Diet was assessed by 4-d dietary records and compliance by plasma alkylresorcinols (ARs)., Results: Fifty-two children (95%) with a BMI z-score of 1.5 ± 0.6 (mean ± standard deviation) completed the study. They consumed 108 ± 38 and 3 ± 2 g/d wholegrain in the WG and RG period, which was verified by a profound difference in ARs (P < 0.001). Compared with RG, WG reduced LDL cholesterol by 0.14 (95% confidence interval: -0.24, -0.04) mmol/L (P = 0.009) and reduced total:high-density lipoprotein cholesterol (P < 0.001) and triacylglycerol (P = 0.048) without altering body composition or other cardiometabolic markers. WG also modulated the abundance of specific bacterial taxa, increased plasma acetate, propionate, and butyrate and fecal butyrate and reduced fatigue with no other effects on gut symptoms., Conclusion: High intake of wholegrain oats and rye reduced LDL cholesterol and triacylglycerol, modulated bacterial taxa, and increased beneficial metabolites in children. This supports recommendations of exchanging refined grain with wholegrain oats and rye among children. This trial was registered at clinicaltrials.gov as NCT04430465., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Our objective was to investigate whether short-chain fatty acids (SCFAs), specifically acetate and butyrate, could prevent vascular dysfunction and elevated blood pressure (BP) in mice with systemic lupus erythematosus (SLE) induced by TLR7 activation using imiquimod (IMQ). Treatment with both SCFAs and dietary fibers rich in resistant starch (RS) or inulin-type fructans (ITF) effectively prevented the development of hypertension and cardiac hypertrophy. Additionally, these treatments improved aortic relaxation induced by acetylcholine and mitigated vascular oxidative stress. Acetate and butyrate treatments also contributed to the maintenance of colonic integrity, reduced endotoxemia, and decreased the proportion of helper T (Th)17 cells in mesenteric lymph nodes (MLNs), blood, and aorta in TLR7-induced SLE mice. The observed changes in MLNs were correlated with increased levels of GPR43 mRNA in mice treated with acetate and increased GPR41 levels along with decreased histone deacetylase (HDAC)- 3 levels in mice treated with butyrate. Notably, the effects attributed to acetate, but not butyrate, were nullified when co-administered with the GPR43 antagonist GLPG-0974. T cell priming and differentiation into Th17 cells in MLNs, as well as increased Th17 cell infiltration, were linked to aortic endothelial dysfunction and hypertension subsequent to the transfer of faecal microbiota from IMQ-treated mice to germ-free (GF) mice. These effects were counteracted in GF mice through treatment with either acetate or butyrate. To conclude, these findings underscore the potential of SCFA consumption in averting hypertension by restoring balance to the interplay between the gut, immune system, and vascular wall in SLE induced by TLR7 activation., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Fatigue among patients with head and neck cancer (HNC) has been associated with higher inflammation. Short-chain fatty acids (SCFAs) have been shown to have anti-inflammatory and immunoregulatory effects. Therefore, this study aimed to examine the association between SCFAs and fatigue among patients with HNC undergoing treatment with radiotherapy with or without concurrent chemotherapy. Plasma SCFAs and the Multidimensional Fatigue Inventory-20 were collected prior to and one month after the completion of treatment in 59 HNC patients. The genome-wide gene expression profile was obtained from blood leukocytes prior to treatment. Lower butyrate concentrations were significantly associated with higher fatigue (p = 0.013) independent of time of assessment, controlling for covariates. A similar relationship was observed for iso/valerate (p = 0.025). Comparison of gene expression in individuals with the top and bottom 33% of butyrate or iso/valerate concentrations prior to radiotherapy revealed 1,088 and 881 significantly differentially expressed genes, respectively (raw p < 0.05). The top 10 Gene Ontology terms from the enrichment analyses revealed the involvement of pathways related to cytokines and lipid and fatty acid biosynthesis. These findings suggest that SCFAs may regulate inflammatory and immunometabolic responses and, thereby, reduce inflammatory-related symptoms, such as fatigue., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)

In this study, we synthesized and incorporated chitosan nanoparticles (Cs) into polyhydroxy butyrate (PHB) electrospun scaffolds for cartilage tissue engineering. The Cs nanoparticles were synthesized via an ionic gel interaction between Cs powder and tripolyphosphate (TPP). The mechanical properties, hydrophilicity, and fiber diameter of the PHB scaffolds with varying concentrations of Cs nanoparticles (1-5 wt%) were evaluated. The results of these evaluations showed that the scaffold containing 1 wt% Cs nanoparticles (P1Cs) was the optimum scaffold, with increased ultimate strength from 2.6 to 5.2 MPa and elongation at break from 5.31 % to 12.6 %. Crystallinity, degradation, and cell compatibility were also evaluated. The addition of Cs nanoparticles decreased crystallinity and accelerated hydrolytic degradation. MTT assay results showed that the proliferation of chondrocytes on the scaffold containing 1 wt% Cs nanoparticles were significantly higher than that on pure PHB after 7 days of cultivation. These findings suggest that the electrospun P1Cs scaffold has promising potential as a substrate for cartilage tissue engineering applications. This combination offers a promising approach for the fabrication of biomimetic scaffolds with enhanced mechanical properties, hydrophilicity, and cell compatibility for tissue engineering applications., Competing Interests: Declaration of competing interest All authors declare that there is no financial/commercial conflicts of interest., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Medium chain fatty acids (MCFA) generation is attracting growing interest due to fossil fuel depletion. To promote the production of MCFA, especially caproate, hydrochloric acid pretreated activated carbon (AC) was introduced into chain elongation fermentation. In this study, the role of pretreated AC on caproate production was investigated using lactate and butyrate as electron donor and electron acceptor, respectively. The results showed that AC did not improve the chain elongation reaction at beginning but promoted the caproate production at later stage. The addition of 15 g/L AC facilitated reactor reaching the peak of caproate concentration (78.92 mM), caproate electron efficiency (63.13%), and butyrate utilization rate (51.88%). The adsorption experiment revealed a positive correlation between the adsorption capacity of pretreated AC and the concentration as well as the carbon chain length of carboxylic acids. Moreover, the adsorption of undissociated caproate by pretreated AC contributed to a mitigated toxicity towards microorganisms, thereby facilitating the production of MCFA. Microbial community analysis revealed an increasing enrichment of key functional chain elongation bacteria, including Eubacterium, Megasphaera, Caproiciproducens, and Pseudoramibacter, but a suppression on acrylate pathway microorganism Veillonella, as the dosage of pretreated AC increasing. The findings of this study demonstrated the substantial impact of the adsorption effect of acid-pretreated AC on promoting caproate production, which would aid to the development of more efficient caproate production process., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Background: Obesity has reached epidemic dimensions in recent decades. Bariatric surgery (BS) is one of the most effective interventions for weight loss and metabolic improvement in patients with obesity. Short-chain fatty acids (SCFA) are gut microbiota-derived metabolites with a key role in body weight control and insulin sensitivity. Although BS is known to induce significant changes in the gut microbiota composition, its impact on the circulating levels of certain metabolites produced by the gut microbiota such as SCFA remains poorly understood., Objective: To determine the impact of BS on the circulating SCFA levels in patients with severe obesity., Setting: University hospital., Methods: An observational, prospective study was performed on 51 patients undergoing Roux-en-Y gastric bypass. Plasma samples were collected at baseline (1 day before surgery) and at 6 and 12 months after BS. Plasma SCFA levels were determined by liquid chromatography-mass spectrometry., Results: The results revealed significant changes in the circulating levels of SCFA after BS. A marked increase in propionate, butyrate, isobutyrate, and isovalerate levels and a decrease in acetate, valerate, hexanoate, and heptanoate levels were observed 12 months after BS. Furthermore, the changes in the levels of propionate, butyrate, and isobutyrate negatively correlated with changes in body mass index, while those of isobutyrate correlated negatively with changes in the homeostatic model assessment for insulin resistance index., Conclusion: These results suggest that propionate, butyrate, and isobutyrate levels could be related to weight loss and improved insulin sensitivity in patients with severe obesity after BS., (Copyright © 2023 American Society for Metabolic and Bariatric Surgery. Published by Elsevier Inc. All rights reserved.)

The aim of the present work was the selection of aromatic plant essential oils (EOs) and/or ethanolic extracts (EEs) to prevent the late blowing defect (LBD) of cheese caused by Clostridium spp. EEs resulted more effective than EOs to inhibit dairy-borne Clostridium spp. in vitro. Savory, hyssop, lavender and tarragon EEs, which showed the lowest minimal inhibitory concentration against Clostridium tyrobutyricum, were selected to study the prevention of LBD caused by this bacterium in cheese. Addition of savory and lavender EEs to cheese milk delayed LBD by 2 weeks, but at the end of ripening these cheeses showed similar clostridial vegetative cells counts, spoilage symptoms and propionic, and butyric acids levels than blown control cheese. Tarragon EE, with the highest content in caffeic acid, also delayed LBD by 2 weeks, but it was more effective to inhibit Clostridium, since cheese with tarragon EE showed minor LBD symptoms, lower vegetative cells count and lower concentrations of propionic and butyric acids than the rest of cheeses made with EEs. This fact could be also attributable to the greater number of antimicrobial terpenes (1,8-cineole, 4-terpineol, α-terpineol, isoelemicin, methyl eugenol, and methyl trans-isoeugenol) detected in this cheese. This is the first report on the application of EEs to control C. tyrobutyricum in cheese., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The production of volatile fatty acids (VFA) is gaining momentum due to their central role in the emerging carboxylate platform. Particularly, the production of the longest VFA (from butyrate to caproate) is desired due to their increased economic value and easier downstream processing. While the use of undefined microbial cultures is usually preferred with organic waste streams, the use of defined microbial co-culture processes could tackle some of their drawbacks such as poor control over the process outcome, which often leads to low selectivity for the desired products. However, the extensive experimentation needed to design a co-culture system hinders the use of this technology. In this work, a workflow based on the combined use of mathematical models and wet experimentation is proposed to accelerate the design of novel bioprocesses. In particular, a co-culture consisting of Pediococcus pentosaceus and Megaphaera cerevisiae is used to target the production of high-value odd- and even‑carbon VFA. An unstructured kinetic model was developed, calibrated and used to design experiments with the goal of increasing the selectivity for the desired VFA, which were experimentally validated. In the case of even‑carbon VFA, the experimental validation showed an increase of 38 % in caproate yield and, in the case of enhanced odd‑carbon VFA experiments, the yield of butyrate and caproate diminished by 62 % and 94 %, respectively, while propionate became one of the main end products and valerate yield value increased from 0.007 to 0.085 g valearte per g consumed sugar. The workflow followed in this work proved to be a sound tool for bioprocess design due to its capacity to explore and design new experiments in silico in a fast way and ability to quickly adapt to new scenarios., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Author(s). Published by Elsevier B.V. All rights reserved.)

The report describes an updated, fully automated method for the production of [ 11 C]butyrate, validated for use in clinical studies. A commercially available GE Tracerlab FX M synthesis module was reconfigured to allow for air-free introduction of n-propyl magnesium chloride and to incorporate Sep-Pak cartridges to simplify and shorten the purification process, as compared to purifying the product using traditional HPLC. The method takes 20 min from end-of-bombardment and reliably produces injectable doses of [ 11 C]butyrate (8029 ± 1628 MBq (217 ± 44 mCi), 14 % radiochemical yield based on [ 11 C]CO 2, non-decay corrected) in high radiochemical purity (>97 %), n = 3. With radiotracer in hand, PET scans of rats confirmed uptake of the radiopharmaceutical in the brain. Rat biodistribution data was obtained and used in conjunction with OLINDA software to determine an estimated human total body effective dose of 3.20 × 10 -3  mSv/MBq (1.19 × 10 -2  rem/mCi), along with preliminary rodent PET imaging that confirmed brain uptake. Lastly, our first human [ 11 C]butyrate PET studies using a dynamic bolus injection technique (n = 5), with a graphical Logan analysis using a white matter reference region, confirmed good radiotracer uptake in the brain and with relatively more prominent uptake in the cerebellar hemispheres, vermis, cingulum cortex and the thalami., Competing Interests: Declaration of competing interest The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. Given their role as Editor, Peter J. H. Scott had no involvement in the peer-review of this article and has no access to information regarding its peer-review., (Copyright © 2023. Published by Elsevier Inc.)

The addition of carbon (C) substrate often modifies the rate of soil organic matter (SOM) decomposition. This is known as the priming effect. Nitrous oxide (N 2 O) emissions from soil are also linked to C substrate dynamics; however, the relationship between the priming effect and N 2 O emissions from soil is not understood. This study aimed to investigate the effects of C and N substrate addition on the linkages between SOM priming and N 2 O emissions. We applied 13 C-labelled substrates (acetate, butyrate, glucose; 80 μg C g -1 ), with water as a control, and 15 N-labelled N (300 μg N g -1 soil, potassium nitrate) to three different soils, and, after 3 days, we measured the effects on the priming of SOM and sources of N 2 O emission. Carbon substrate addition increased both CO 2 - and SOM-derived N 2 O emissions in the presence of exogenous N. Emissions of CO 2 and N 2 O from soils with added glucose (mean ± standard deviation, 0.73 ± 0.13 μmol m -2 s -1 and 21.4 ± 12.1 mg N m -2 h -1 ) were higher (p < 0.05) than those from soils treated with acetate (0.64 ± 0.11 μmol m -2 s -1 and 10.9 ± 6.5 mg N m -2 h -1 ) or butyrate (0.61 ± 0.11 μmol m -2 s -1 and 11.0 ± 6.6 mg N m -2 h -1 ), respectively. Acetate addition induced a stronger (p < 0.05) priming effect on soil C (0.07 ± 0.09 μmol C m -2 s -1 ) than that for glucose (0.02 ± 0.10 μmol C m -2 s -1 ), while butyrate addition resulted in negative priming (-0.09 ± 0.05 μmol C m -2 s -1 ). SOM-derived N 2 O emissions were relatively low from soils with butyrate addition (1.4 ± 1.5 mg N m -2 h -1 ) compared with acetate (2.9 ± 2.3 mg N m -2 h -1 ) or glucose (9.2 ± 4.5 mg N m -2 h -1 ). There was no clear relationship between the priming effect and SOM-derived N 2 O emissions. The observed priming effect related to the potential electron donor supply of the C substrates was not observed. There is a need to further examine the role of soil priming in relation to soil N 2 O emissions., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Waste valorisation via biological production of widely used in the industry medium chain carboxylates (MCCs) via open culture fermentation (OCF) could be a promising alternative to the commonly used anaerobic digestion. Lactate-rich waste streams are considered as valuable substrates for carboxylate chain elongation (CE), however, there are certain limitations related to the production efficiency. Acetate produced and accumulated in the acetogenesis plays an important role in CE, i.e. acetate is elongated to butyrate and then to caproate which is most popular MCC. Henceforth, it was investigated whether the ratio of lactate to acetate (L:A) affected carboxylates yields and product distribution in the lactate-based CE in OCF. The tested L:A ratios influenced carboxylates selectivity in batch trials. In the ones with lactate as the sole carbon source, propionate production was predominant but when a higher relative acetate concentration was used, the production of butyrate and CE to caproate was favored. The co-utilization of lactate and acetate in a continuous process increased the production of butyrate and caproate compared to the phase with lactate as the sole carbon source, however, controlling the relative concentration of lactate and acetate during co-utilization was not an effective strategy for increasing caproate production. 16S rRNA gene amplicon reads mapping to Caproiciproducens were the most abundant in samples collected throughout the continuous processes regardless of the L:A ratios., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)

Following allogeneic hematopoietic cell transplantation (allo-HCT), the gastrointestinal (GI) tract is frequently affected by acute graft-versus-host disease (aGVHD), the pathophysiology of which is associated with a dysbiotic microbiome. Since microbial composition varies along the length of the GI tract, the authors hypothesized that microbiome features correlate with the pattern of organ involvement after allo-HCT. We evaluated 266 allo-HCT recipients from whom 1303 stool samples were profiled by 16S ribosomal gene sequencing. Patients were classified according to which organs were affected by aGVHD. In the 20 days prior to disease onset, GVHD patients had lower abundances of members of the class Clostridia, lower counts of butyrate producers, and lower ratios of strict-to-facultative (S/F) anaerobic bacteria compared with allograft recipients who were free of GVHD. GI GVHD patients showed significant reduction in microbial diversity preonset. Patients with lower GI aGVHD had lower S/F anaerobe ratios compared with those with isolated upper GI aGVHD. In the 20 days after disease onset, dysbiosis was observed only in GVHD patients with GI involvement, particularly those with lower-tract disease. Importantly, Clostridial and butyrate-producer abundance as well as S/F anaerobe ratio were predictors of longer overall survival; higher abundance of butyrate producers and higher S/F anaerobe ratio were associated with decreased risk of GVHD-related death. These findings suggest that the intestinal microbiome can serve as a biomarker for outcomes of allo-HCT patients with GVHD., (© 2022 by The American Society of Hematology.)

Bioelectrochemical systems were proposed as a promising approach for the efficient valorization of biomass into 6-8 carbon atom medium-chain fatty acids (MCFAs), the precursors for high value-added chemicals or renewable energy, via acetyl-CoA-mediated chain elongation (CE). To achieve CE processes, exogenous electron donors (EDs), e.g., ethanol or lactic acid, were normally prerequisites. This research built a microbial electrolysis cell (MEC) for MCFAs biosynthesis from acetate without exogenous EDs addition. A wide range of applied voltages (0.6-1.2 V) was first employed to investigate the bioelectrocatalyzing response. The results show that caproate and butyrate were the main products formed from acetate under different applied voltages. Maximum caproate concentration (501 ± 12 mg COD/L) was reached at 0.8 V on day 3. Under this applied voltage, hydrogen partial pressure stabilized at about 0.1 bar, beneficial for MCFA production. Electron and carbon balances revealed that the electron-accepting capacity achieved 32% at 0.8 V, showing the highest interspecies electron transfer efficiency. Most of the carbon was recovered in the form of caproate (carbon loss was 9%). MiSeq sequencing revealed Rhodobacter and Clostridium&#95;sensu&#95;stricto playing the crucial role in the biosynthesis of caproate, while Acetobacterium, Acetoanaerobium, and Acetobacter represented the main ED contributors. Four available flora, i.e., homo-acetogen, anaerobic fermentation bacteria, electrode active bacteria, and nitrate-reducing bacteria, interacted and promoted caproate synthesis by molecular ecological network analysis., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Volatile fatty acids (VFAs) derived from biomass are considered to be economical and environmentally friendly feedstocks for microbial fermentation. Converting VFAs to polyhydroxyalkanoate (PHA) could reduce the substrate cost and provide an economically viable route for the commercialization of PHA. The halophilic bacteria Salinivibrio spp. TGB4 and TGB19, newly isolated from salt fields, were found to accumulate poly-3-hydroxybutyrate (PHB) using acetate or butyrate as the substrate. Both strains exhibited considerable cell growth (OD 600 of ~8) even at acetate concentration of 100 g/L. In shake flask cultures, TGB4 produced PHB titers of 0.90 and 1.34 g/L, while TGB19 produced PHB titers of 0.25 and 2.53 g/L with acetate and butyrate, respectively. When acetate and butyrate were both applied, PHB production was significantly increased, and the PHB titer of TGB4 and TGB19 reached 6.14 and 6.84 g/L, respectively. After optimizing the culture medium, TGB19 produced 8.42 g/L PHB, corresponding to 88.55 wt% of cell dry weight. During fed-batch cultivation, TGB19 produced a PHB titer of 53.23 g/L. This is the highest reported PHB titer using acetate and butyrate by pure microbial cultures and would provide promising hosts for the industrial production of PHA from VFAs., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Aims: Short-chain fatty acids (SCFAs) are produced by gut microbiota from dietary fiber. Since absorbed SCFAs could be introduced into the tricarboxylic acid (TCA) cycle in host cells, the relationships between SCFAs and TCA cycle intermediates might influence to energy metabolism in the human body. For this reason, information on profile changes between SCFAs and TCA cycle intermediates could help unveil pathological mechanisms of gastric cancer., Main Methods: A gas chromatography-tandem mass spectrometry (GC-MS/MS) method was developed to simultaneously determine SCFAs and TCA cycle intermediates in human plasma from patients with chronic superficial gastritis (CSG), intestinal metaplasia (IM), and gastric cancer. We applied a tetra-alkyl ammonium pairing method to prevent loss of volatile SCFAs and base decarboxylation of TCA cycle intermediates during sample preparation. To assess gastric diseases, metabolic alterations of SCFAs and TCA cycle intermediates in human plasma with gastric disorders were analyzed by their plasma levels., Key Findings: Significantly different metabolic alterations based on the plasma levels of SCFAs and TCA cycle intermediates were investigated in cancer metabolic pathways. Not only propionate and butyrate, mainly produced by gut microbiota, were significantly decreased, but also cis-aconitate, α-ketoglutarate, and fumarate were significantly increased in plasma with IM or gastric cancer, compared to CSG. Further, based on ratios of product to precursor, three metabolic pathways (succinate/propionate, succinate/α-ketoglutarate, and cis-aconitate/citrate) were supposed to be distorted between gastric diseases., Significance: In conclusion, propionate, cis-aconitate, α-ketoglutarate, and fumarate could be used to assess the progression of gastric cancer., Competing Interests: Declaration of competing interest The authors declare that there are no conflicts of interest., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Background and Aims: Berry (poly)phenol consumption has been associated with cardioprotective benefits, however little is known on the role the gut microbiome may play on such health benefits. Our objective was to investigate the effects of aronia berry (poly)phenol consumption on cardiometabolic health and gut microbiome richness and composition in prehypertensive middle-aged men and women., Methods: A total of 102 prehypertensive participants were included in a parallel 12-week randomized double-blind placebo-controlled trial. Volunteers were randomly allocated to daily consume an encapsulated (poly)phenol-rich aronia berry extract (Aronia, n = 51) or a matched maltodextrin placebo (Control, n = 51). Blood pressure (BP) and arterial function (office and 24 h), endothelial function (measured as flow-mediated dilation), serum biochemistry (including blood lipids), plasma and urine (poly)phenol metabolites as well as gut microbiome composition through shotgun metagenomic sequencing were monitored over the study period. Relationships between vascular outcomes, (poly)phenol metabolites and gut microbiome were investigated using an integrated multi-levels approach., Results: A significant improvement in arterial indices measured as augmentation index (AIx) and pulse wave velocity (PWV) was found in the Aronia compared to Control group (awake Δ PWV = -0.24 m/s; 95% CI: -0.79, -0.01 m/s, P < 0.05; 24 h peripheral Δ AIx = -6.8; -11.2, -2.3, %, P = 0.003; 24 h central Δ AIx = -3.3; -5.5, -1.0, %, P = 0.006). No changes in BP, endothelial function or blood lipids were found following the intervention. Consumption of aronia (poly)phenols led to a significant increase in gut microbiome gene richness and in the abundance of butyrate-producing species such as Lawsonibacter asaccharolyticus and Intestinimonas butyriciproducens species, compared to Control group. Results from an approach including metabolomic, metagenomic and clinical outcomes highlighted associations between aronia-derived phenolic metabolites, arterial stiffness, and gut microbiome., Conclusions: Aronia berry (poly)phenol consumption improved arterial function in prehypertensive middle-aged individuals, possibly via modulation of gut microbiome richness and composition based on the associations observed between these parameters., Clinical Trial Registry: The National Institutes of Health (NIH)-randomized trial records held on the NIH ClinicalTrials.gov website (NCT03434574). Aronia Berry Consumption on Blood Pressure., Competing Interests: Conflict of interest EF, MLecomte, PFB are employees of Naturex SA, part of Givaudan. MLS, YX, FAG, DK, CD, UK, SA, JKC and ARM were supported by a collaborative research agreement with Naturex SA, part of Givaudan. FPO and MLaiola were supported by a service agreement with Naturex SA, part of Givaudan., (Copyright © 2022 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Background: SCFAs have immune-modulating effects in animal models of disease. However, there is limited evidence that this may occur in humans., Objectives: This study aimed to determine the effects of increased exposure to SCFAs via dietary manipulation on colonic fermentation and adaptive immune cells., Methods: Twenty healthy young adults (18-45 y of age) underwent a blinded randomized crossover dietary intervention, consuming a high-SCFA diet and a matched low-SCFA diet for 21-d with a 21-d washout in between. SCFAs were provided through resistant starch, inulin, and apple cider vinegar. Blood and 3-d total fecal output were collected at baseline and at the end of each diet. GC was used to measure fecal and plasma SCFA. Flow cytometry was used for peripheral blood immunophenotyping., Results: According to a paired samples Wilcoxon test, the high-SCFA diet was associated with significantly higher fecal SCFA concentrations [median (IQR); 86.6 (59.0) compared with 75.4 (56.2) µmol/g, P = 0.02] and lower fecal ammonia concentrations [26.2 (14.7) compared with 33.4 (18.5) µmol/g, P = 0.04] than the low-SCFA diet. Plasma propionate [9.87 (12.3) compared with 4.72 (7.6) µmol/L, P = 0.049] and butyrate [2.85 (1.35) compared with 2.02 (1.29) µmol/L, P = 0.03] were significantly higher after the high-SCFA diet than after the low-SCFA diet. Blood total B cells [184 (112) compared with 199 (143) cells/µL, P = 0.04], naive B cells [83 (66) compared with 95 (89) cells/µL, P = 0.02], Th1 cells [22 (19) compared with 29 (16) cells/µL, P = 0.03], and mucosal-associated invariant T cells [62 (83) compared with 69 (114) cells/µL, P = 0.02] were significantly lower after the high-SCFA diet than the low-SCFA diet., Conclusions: Increasing colonic and peripheral blood SCFA has discrete effects on circulating immune cells in healthy humans following 3-wk intervention. Further studies (e.g., in patients with inflammatory disease) are necessary to determine whether 1) these changes have immunomodulatory effects, 2) they are therapeutically beneficial, and 3) prolonged intake might be required. This trial is registered at the Australian New Zealand Clinical Trials Registry as ACTRN12618001054202., (Copyright © 2022 American Society for Nutrition.)

Background: The Mediterranean diet is associated with the prevention of diabetes, cardiovascular disease, and cancer, all of which are linked to intestinal barrier impairment., Objectives: Here, we hypothesize that the Mediterranean diet, possibly via the induction of short-chain fatty acids (SCFAs), improves intestinal barrier integrity. Furthermore, we aim to establish novel personalized nutrition advice based on machine learning algorithms., Methods: We studied 260 women with intestinal barrier impairment. The women were allocated to follow either a Mediterranean diet or a control diet for 3 mo. Intestinal permeability was assessed by measuring lipopolysaccharide binding protein (LBP) in plasma and zonulin in feces. SCFA concentrations were analyzed in feces. Bi- and multivariate analyses and machine learning algorithms (random forest classification) were conducted., Results: Particularly in the intervention group, adherence to the Mediterranean diet increased, whereas plasma LBP and fecal zonulin concentrations decreased (all q < 0.001 for the intervention group, all q < 0.1 for control group). In the intervention group, fecal SCFA concentrations increased (propionate + 19%; butyrate + 44%; both q < 0.001). Multivariate analyses showed that adherence to the Mediterranean diet was associated with SCFA concentrations (all q < 0.001) and inversely associated with LBP and zonulin concentrations (all q < 0.02). Mediation analyses identified propionate and butyrate as the key mechanistic link between diet and intestinal permeability integrity. Accordingly, using baseline SCFA data, we could predict the effect of the Mediterranean diet on intestinal permeability using a machine learning algorithm (receiver operating characteristic AUC: 0.78-0.96)., Conclusions: Our data suggest that SCFAs are key mediators for the relation between diet and gut health. Assessment of SCFAs may form a basis for personalized nutrition in future clinical care. These results need to be verified in larger studies powered for this purpose, comprising different study populations. The trial was registered at clinicaltrials.gov as NCT02087592 and NCT02516540., (© The Author(s) 2022. Published by Oxford University Press on behalf of the American Society for Nutrition.)

Background: Tree nuts have glucoregulatory effects and influence gut microbiota composition. The effect of peanuts on the microbiota has not been investigated., Objectives: The aim was to examine the effect of 28 g/d of peanuts for 6-wks, compared to an isocaloric lower-fat higher-carbohydrate (LFHC) snack, on gut microbiota composition. A secondary aim was to identify functional and active compositional differences in a subset of participants using metatranscriptomics., Methods: In a randomized, crossover trial, 50 adults (48% female; 42 ± 15 y; BMI 28.3 ± 5.6 kg/m 2 ; plasma glucose 100 ± 8 mg/dL) consumed 28 g/d of dry roasted, unsalted, peanuts (164 kcal; 11% E carbohydrate, 17% E protein, 73% E fat, and 2.4 g fiber) or a LFHC snack (164 kcal; 53% E carbohydrate, 17% E protein, 33% E fat, and 3 g fiber) for 6-wk (4-wk washout period). Gut bacterial composition was measured using 16S rRNA sequencing in the whole cohort. Exploratory metatranscriptomic analyses were conducted on a random subset (n = 24) of samples from the Peanut condition., Results: No between-condition differences in α- or β- diversity were observed. Following peanut intake, Ruminococcaceae were significantly more abundant [Linear discriminant analysis score (LDA) = 2.8; P = 0.027)] compared to LFHC. Metatranscriptomics showed increased expression of the K03518 (aerobic carbon-monoxide dehydrogenase small subunit) gene following peanut intake (LDA = 2.0; P = 0.004) and Roseburia intestinalis L1-82 was identified as a contributor to the increased expression., Conclusion: An increased abundance of Ruminococcaceae was observed following consumption of 28 g/d of peanuts in adults with elevated fasting glucose after 6-wks. Metatranscriptomics revealed increased expression of the K03518 gene. These results suggest peanut intake enriches a known butyrate producer and the increased expression of a gene implicated in butyrate production adds further support for peanut-induced gut microbiome modulation. NCT: 03654651., (Copyright © 2022 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Cold exposure has been recognized as an important risk factor for hypertension, and altered gut microbiota has been reported to be associated with hypertension. We hypothesized that there is a plausible relationship between gut microbiota and cold-induced hypertension (CIH). Therefore, we explored the potential link between the gut microbiota and its metabolites with CIH. Male Sprague-Dawley (SD) rats were randomly divided into the normal temperature group (NT, 20 ± 2 °C) and the cold exposure group (CE, 4 ± 1 °C), and faecal bacteria cross-transplantation was performed after six weeks. We analyzed the gut microbiota of rats using the 16S rDNA sequence and measured the blood pressure of rats and the content of short-chain fatty acids in rat faeces. After six weeks of cold exposure, the CIH rat model was successfully established. The cold exposure reduced the diversity of the gut microbiota, increased the abundance of potentially pathogenic and conditionally pathogenic bacteria (e.g., Quinella, Rothia, and Senegalimassilia genera), and reduced the abundance of beneficial bacteria (e.g., Lactobacillus genus) and butyric acid-producing bacteria (e.g., Lachnospiraceae UCG-008 and Ruminococcaceae UCG-013 genera). Faecal bacteria cross-transplantation altered gut microbiota composition and regulated blood pressure levels. The NT group rats transplanted with CIH rats' faecal bacteria were enriched with certain conditional pathogenic bacteria such as Prevotellaceae UCG-003 genus. The CIH rats transplanted with faecal bacteria from the NT group rats were enriched with beneficial bacteria such as Bacteroides genus. In addition, we found a significant reduction in butyric acid levels in CIH rats, which may be related to the increase in blood pressure. In conclusion, CIH is associated with altered gut microbiota and reduced butyric acid. Our findings provide novel insights for the prevention and treatment of CIH by modulating the gut microbiota through supplementation of beneficial bacteria/butyrate., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022. Published by Elsevier B.V.)

Microbial electrosynthesis (MES) is a renewable energy platform capable of reducing the carbon footprint by converting carbon dioxide/bicarbonate to useful chemical commodities. However, the development of feasible electrode structures, inefficient current densities, and the production of unfavorable electrosynthesis products remain a major challenge. To this end, a three-dimensional (3D) macroporous sponge coated with a carbon nanotube/MXene composite (CNT-MXene@Sponge) was evaluated as an MES cathode. The macroporous scaffold, together with intrinsic electrical conductivity, enhanced the charge transfer efficiency and selective microbial enrichment characteristics of the CNT-MXene@Sponge cathode resulted in an average current density of -324 mA m -2 , which was substantially higher than that of the uncoated (-100 mA m -2 ), CNT (-141 mA m -2 ), and MXene (-214 mA m -2 ) coated sponge electrode. The uniform 3D structure and abundant active sites of the coated material facilitated mass diffusion and microbial growth, which produced 1.5 orders of magnitude higher butyrate than the uncoated sponge. The high-throughput sequencing results showed the selective enrichment of electrogenic and butyrate-producing phylum, Firmicutes. These results suggest that the MES performance could be enhanced using the collective features of large-pore network structure, such as better conductivity, improved capacitance, and selective microbial enrichment., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Background and Aims: Despite animal studies revealing a causal link between the gut microbiota and skeletal muscle mass, the role of the gut microbiome and its metabolites in humans having low muscle mass remains unclear., Methods: Eighty-eight subjects older than 65 years were measured for sarcopenia-related parameters, including body composition, grip strength, gait speed and flexibility. Participants were divided into normal muscle mass group (NM, n = 52) and low muscle mass group (LM, n = 36) and fresh fecal samples were collected for metagenome and short chain fatty acids (SCFAs) analysis., Results: The richness and evenness of gut microbiota diversity were significantly decreased in the subjects with low muscle mass, including observed ASVs, Shannon and Chao 1 index. A significant difference of gut microbiota profile was noted between NM group and LM group. The Firmicutes/Bacteroidetes ratio was significantly reduced in the LM group. A significant decrease in the abundance of a SCFA-producer, Marvinbryantia spp., whereas a remarkable enrichment of a flavonoid degrader, Flavonifractor spp., was observed in the LM elders. Comparing with the NM group, the fecal butyrate significantly diminished in the LM group and correlated with skeletal muscle mass index., Conclusions: This is the first study that demonstrates the reduced fecal butyrate in elders with low muscle mass and highlights the associated gut microbiome changes. The identified gut microbial features and fecal butyrate level may serve as potential biomarkers for early detection of sarcopenic patients., Competing Interests: Conflict of interest The authors declare that they have no conflict of interest., (Copyright © 2022 Elsevier Ltd and European Society for Clinical Nutrition and Metabolism. All rights reserved.)

Different combinations of gut health-promoting dietary interventions were tested to support broilers during different stages of Eimeria infection. One-day-old male Ross 308 broilers (n = 720) were randomly assigned to one of 6 dietary treatments, with 6 pens per treatment and 20 birds per pen, for 35 d. At 7 d of age (d7), all birds were inoculated with 1000, 100, and 500 sporulated oocysts of E. acervulina, E. maxima, and E. tenella, respectively. A 4-phase feeding schedule was provided. The dietary treatments (TRT) 1 to 4 included the basal diet supplemented with multispecies probiotics from d0 to 9 and coated butyrate and threonine from d28 to 35 but received four different combinations of prebiotics and phytochemicals from d9 to 18 and d18 to 28. The basal diet for the positive control (PC, TRT5) included diclazuril as a anticoccidial. The negative control (NC, TRT6) contained no anticoccidial. Performance was assessed for each feeding phase, and oocyst output, Eimeria lesion scores, cecal weight, litter quality, and footpad lesions were assessed at d14, d22, d28, and d35. Body weight gain (BWG) and feed intake (FI) were not affected by dietary treatment. PC broilers had the best feed conversion ratio (FCR) of all treatments from d0 to 35 (P < 0.001). None of the dietary treatments resulted in better litter quality or reduced footpad lesions compared to the PC. Moreover, the PC was most effective in reducing oocyst output and lesion scores compared to all other treatments. However, broilers that received the multispecies probiotics (d0 to 9), saponins (d9 to 18), saponins, artemisin, and curcumin (d18 to 28), and coated butyrate and threonine (d28 to 35) had the best FCR (P < 0.001) and lowest oocyst output and lesion scores compared to other dietary treatments. This study suggests that although the tested compounds did not perform as well as the anticoccidial, when applied in the proper feeding period, they may support bird resilience during coccidiosis infection., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)

The intestinal microbiota is essential for the fermentation of dietary fiber into short-chain fatty acids (SCFA) such as butyrate, acetate, and propionate. SCFAs can bind to the G-protein-coupled receptors GPR43 and GPR109A (HCAR2), with varying affinities to promote cellular effects in metabolism or changes in immune function. We explored the role of GPR109A as the main receptor for butyrate in mouse models of allogeneic hematopoietic cell transplantation (allo-HCT) and graft-versus-host disease (GVHD). Deletion of GPR109A in allo-HCT recipients did not affect GVHD, but transplantation of T cells from GPR109A knockout (KO) (Gpr109a-/-) mice into allo-HCT recipient mice significantly reduced GVHD morbidity and mortality compared with recipients of wild-type (WT) T cells. Recipients of Gpr109a-/- T cells exhibited less GVHD-associated target organ pathology and decreased proliferation and homing of alloreactive T cells to target tissues. Although Gpr109a-/- T cells did not exhibit immune deficits at a steady state, following allo-activation, Gpr109a-/- T cells underwent increased apoptosis and were impaired mitochondrial oxidative phosphorylation, which was reversible through antioxidant treatment with N-acetylcysteine (NAC). In conclusion, we found that GPR109A expression by allo-activated T cells is essential for metabolic homeostasis and expansion, which are necessary features to induce GVHD after allo-HCT., (© 2022 by The American Society of Hematology.)

In the context of "Peak CO 2 emissions & Carbon neutrality", H 2 energy, as the green and clean energy, will make an important contribution to the carbon emission reduction and carbon neutralization. Bio-H 2 production from organic wastewater achieved not only pollutants removal, but also the H 2 energy recovery and carbon emission reduction. In this study, a maltose-preferring producer of Clostridium butyricum NH-02 was investigated for the potential and performance of bio-H 2 production from brewery wastewater in batch and semi-continuous fermentation. Appropriate initial pH 7.0 and organic loading of 21,173 mg/L chemical oxygen demand (COD) (2670 mg/L reducing sugar (RS)) stimulated the batch H 2 fermentation efficiency with a maximum H 2 yield of 1.89 mol-H 2 /mol-RS and cumulative H 2 production of 479.3 mL/L. Comparing to the batch fermentation, semi-continuous fermentation showed significant improvement in H 2 productivity and yield. The maximum cumulative H 2 yield of 5.21 mol-H 2 /mol-RS and production of 254.78 mL were obtained with the optimal hydraulic retention time (HRT) at 47 h after a 120 h fermentation. This study demonstrated the potential of H 2 production from brewery wastewater with C. butyricum, and a great improvement in H 2 production in semi-continuous fermentation., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Background & Aims: Although long-chain omega-3 fatty acids (LCn-3FAs) regulate inflammatory pathways of relevance to non-alcoholic steatohepatitis (NASH), their susceptibility to peroxidation may limit their therapeutic potential. We compared the metabolism of eicosapentaenoic acid (EPA) with an engineered EPA derivative (icosabutate) in human hepatocytes in vitro and their effects on hepatic glutathione metabolism, oxidised lipids, inflammation, and fibrosis in a dietary mouse model of NASH, and in patients prone to fatty liver disease., Methods: Oxidation rates and cellular partitioning of EPA and icosabutate were compared in primary human hepatocytes. Comparative effects of delayed treatment with either low- (56 mg/kg) or high-dose (112 mg/kg) icosabutate were compared with EPA (91 mg/kg) or a glucagon-like peptide 1 receptor agonist in a choline-deficient (CD), L-amino acid-defined NASH mouse model. To assess the translational potential of these findings, effects on elevated liver enzymes and fibrosis-4 (FIB-4) score were assessed in overweight, hyperlipidaemic patients at an increased risk of NASH., Results: In contrast to EPA, icosabutate resisted oxidation and incorporation into hepatocytes. Icosabutate also reduced inflammation and fibrosis in conjunction with a reversal of CD diet-induced changes in the hepatic lipidome. EPA had minimal effect on any parameter and even worsened fibrosis in association with depletion of hepatic glutathione. In dyslipidaemic patients at risk of NASH, icosabutate rapidly normalised elevated plasma ALT, GGT and AST and reduced FIB-4 in patients with elevated ALT and/or AST., Conclusion: Icosabutate does not accumulate in hepatocytes and confers beneficial effects on hepatic oxidative stress, inflammation and fibrosis in mice. In conjunction with reductions in markers of liver injury in hyperlipidaemic patients, these findings suggest that structural engineering of LCn-3FAs offers a novel approach for the treatment of NASH., Lay Summary: Long-chain omega-3 fatty acids are involved in multiple pathways regulating hepatic inflammation and fibrosis, but their susceptibility to peroxidation and use as an energy source may limit their clinical efficacy. Herein, we show that a structurally modified omega-3 fatty acid, icosabutate, overcame these challenges and had markedly improved antifibrotic efficacy in a mouse model of non-alcoholic steatohepatitis. A hepatoprotective effect of icosabutate was also observed in patients with elevated circulating lipids, in whom it led to rapid reductions in markers of liver injury., Competing Interests: Conflict of interest NorthSea Therapeutics BV acquired the commercial rights for icosabutate. DS and JK are paid consultants and have stock options in NorthSea Therapeutics. DF and TS are employees of NorthSea Therapeutics BV but had no role in data collection. All other authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. Please refer to the accompanying ICMJE disclosure forms for further details., (Copyright © 2021. Published by Elsevier B.V.)

In addition to offering methionine, 2-hydroxy-4-methylthiobutyric acid (HMTBa) is also an organic acid and shows excellent bacteriostasis. Therefore, 3 experiments were conducted to determine the influence of drinking water supplemented HMTBa in combination with acidifier on performance, intestinal development, and microflora in broilers. The addition of different concentration (0.02-0.20%) of the blend of HMTBa and other acids significantly reduced the pH of water and exerted antimicrobial activity in dose-dependent manner in vitro. The outcomes from animal trial consisting of the drinking water with blended acidifier at 0.00, 0.05, 0.10, 0.15, and 0.20% indicated that the water with 0.15 or 0.20% acidifier resulted in linear and quadratic higher body weight at 42 d, gain and water consumption during 1 to 42 d (P < 0.05). In experiment 3, responding to graded blended acidifier in drinking water, birds receiving 0.10, 0.15, and 0.20% acidifier decreased the internal pH of gastrointestinal tract and muscle, and exhibited increased duodenal weight, length, villus high, and the ratio of villus high to crypt depth. Drinking water with 0.2% blended acidifier increased the abundance of probiotics (Bacteroidaceae, Ruminococcaceae, and Lachnospiraceae) and decreased the account of pathogenic bacteria such as Desulfovibrionaceae. Alternations in gut microflora were closely related to the metabolism of carbohydrate, amino acid, and vitamins. These findings, therefore, suggest that drinking water with 0.10 to 0.13% the combination HMTBa with acidifier might benefit to intestinal development and gut microbiota, and the subsequent produce a positive effect on the performance of broilers., (Copyright © 2021 The Authors. Published by Elsevier Inc. All rights reserved.)

Several species of novel marine bacteria from the genus Marinobacterium, including M. nitratireducens, M. sediminicola, and M. zhoushanense were found to be capable of producing polyhydroxyalkanoates (PHA) using sugars and volatile fatty acids (VFAs) as the carbon source. M. zhoushanense produced poly-3-hydroxybutytate (PHB) from sucrose, achieving a product titer and PHB content of 2.89 g/L and 64.05 wt%, respectively. By contrast, M. nitratireducens accumulated 3.38 g/L PHB and 66.80 wt% polymer content using butyrate as the substrate. A third species, M. sediminicola showed favorable tolerance to propionate, butyrate, and valerate. The use of 10 g/L valerate yielded 3.37 g/L poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV), with a 3-hydroxyvalerate (3 HV) monomer content of 94.75 mol%. Moreover, M. sediminicola could be manipulated to produce PHBV with changeable polymer compositions by feeding different mixtures of VFAs. Our results indicate that M. sediminicola is a promising halophilic bacterium for the production of PHA., (Copyright © 2021 Elsevier B.V. All rights reserved.)