The composition of intestinal microbial communities plays a crucial role in maintaining immune homeostasis, influencing both innate and adaptive immune responses. Growing evidence indicates that bidirectional communication between gut bacteria and host immune cells contributes to the development of autoimmune diseases. Disruptions in microbial diversity, known as dysbiosis, are linked to an increased susceptibility to autoimmune disorders such as rheumatoid arthritis (RA), multiple sclerosis (MS), and lupus erythematosus. This review examines the mechanistic connections between microbial dysregulation and abnormal immune activation, focusing on key signaling pathways. Pathways such as nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), Janus kinase/signal transducers and activators of transcription (JAK/STAT), and Toll-like receptor (TLR) networks act as immunological gatekeepers, and their dysregulation-induced by microbial metabolites or shifts in microbial composition-can lead to chronic inflammation and the breakdown of self-tolerance. Additionally, bacterial fermentation products, including short-chain fatty acids (SCFAs), exert immunomodulatory effects by influencing T-cell differentiation and cytokine profiles. Emerging therapeutic strategies targeting microbial restoration, such as precision probiotics, microbiota transplantation, and tailored nutritional interventions, aim to restore immune balance. This review underscores the gut microbiota as a dynamic regulator of immune signaling.

<i>Russula vinosa</i> Lindblad is a traditional food and medicine resource with potential application in the field of functional foods, which is rich in polyphenolic compounds. In this study, the <i>R. vinosa</i> Lindblad was mixed fermented with <i>Saccharomyces boulardii</i> and <i>Lactobacillus lactis</i> and the antioxidant activity, composition and application in functional bread of the ethanol extract of the mixed bacterial fermentation product (EMFP) were investigated. The results showed that after fermentation, 186 new compounds appeared in EMFP, including organic acids and phenolic acids. The addition of EMFP significantly enhanced the quality and antioxidant activity of the bread, including reduction of hardness and chewiness, enhancement of resilience, and improvement of free radical scavenging activity and total reducing power. Furthermore, the EMFP addition affected color, odor, and texture indicators of the bread, and negatively affected at higher additions. 0.5% EMFP addition might be appropriate to positively affect the quality, sensory evaluation, and antioxidant activity of the bread. This study investigated the positive effects of mixed fermentation on the active ingredients of the <i>R. vinosa</i> Lindblad, providing a reference for the practical application of EMFP in the field of functional foods.

Background:Enhancing the nutritional components of milk would improve its quality, especially ewe milk, due to its economic importance to dairy industry companies.Aim:This study evaluated the effects of adding ground dates (Al-Zuhdi) to the concentrated diet on the productive performance of milk components and bacterial fermentation products in the rumen during the lactation period.Methods:Eighteen ewes, aged 2.0–2.5 years old, in good health, with an average weight of 53.60 ± 7.25 kg B.W, were equally divided into three equal groups. The first group (T1) was fed a basal diet as a control, whereas the ewes of the second group (T2) were fed a basal diet with ground dates at a ratio of concentrated diet. The third group (T3) was fed a basal diet supplemented with 15% ground dates from the concentrate diet.Results:The findings demonstrated that rumen fermentation rates were significantly increased (p < 0.05) when dates were added to the diet of ewes. Supplementation with ground dates raised the levels of NH3-N and total volatile fatty acids. When comparing the treated groups with the control, the relative abundance of rumen fluid’s pH acidity decreased (p < 0.05). A diet supplemented with ground dates significantly increased milk protein and fat percentages in T2 and T3 ewes, but no significant difference was observed in milk lactose and solid non-fat.Conclusion:The current study concluded that incorporating ground dates into nutritional supplements can enhance the activation of rumen microorganisms and provide nutrients to ewes, which is reflected in the enrichment of milk components.

Abstract Breast cancer (BC) is the most common cause of cancer-related death in women. BC affects ∼2.3 million women worldwide, with ∼685, 000 deaths every year. Therefore, a clear understanding of the molecular mechanism(s) associated with BC formation and the development of more effective therapeutic strategies for the prevention and treatment of this disease are sorely needed. The p53 tumor suppressor protein, encoded by the TP53 gene, prevents multicellular organisms from developing cancer. However, p53 mutations occur in almost all tumors, emphasizing its role as a formidable barrier that must be breached to allow oncogenic transformation. p53 is the most frequently mutated gene in BC, especially in triple-negative breast cancer (TNBC). Although p53 mutation has been seen in 18-25% of primary BC, about 80% of TNBC is associated with p53 mutation or inactivation. p53 is a multi-functional protein that controls the cell cycle, cell proliferation, DNA repair, senescence, and apoptosis, thereby blocking the proliferation of damaged cells and preventing tumor formation. Therefore, the development of therapeutic strategies that will rejuvenate wild-type p53 (WTp53) function or target mutant p53 (p53α) should have a high clinical impact on the prevention and treatment of BC. We found that HCAR2 (Hydroxycarboxylic acid receptor 2) or NIACR1 (Niacin receptor 1) or HM74A or GPR109A, a G-protein coupled receptor for the vitamin B3 Niacin (NA), the bacterial fermentation product butyrate (BTR), and the ketone body β-hydroxybutyrate (BHB), activates WTp53 and also destabilizes p53α in BC cells. p53 is an NAD+-dependent molecule, and deficiency of NA, the precursor of NAD+ and HCAR2 agonist impairs p53 function. NA- and BTR-induced HCAR2 activation can enhance p53 function by decreasing cAMP, thereby attenuating the binding of MDM2 to p53 and the resulting proteasomal degradation and inhibiting SIRT1, a p53 deacetylase. Similarly, BHB, an HCAR2 agonist and the primary ketone body in mammals, potentiates p53 function by increasing p53 acetylation. Further, HCAR2 activation induces cytotoxicity in p53α BC cells via inhibition of HSP90-HDAC chaperone, which stabilizes the transcription of p53α in the mammary epithelium. Therefore, HCAR2 activation should have a high clinical translational potential for the prevention and treatment of BC by rejuvenating WTp53 and destabilizing p53α in BC cells. Overall, our study provides preclinical evidence to show the functional implication of ketogenic signaling in the regulation of p53 and its relevance to BC prevention and treatment. Citation Format: SONIA BATAN, Nanditi Thangaraju, Jabuneesa Khanom, Kunal Kumar, Snigdha Ganjikunta, Subash Sundaram, Nikhil Patel, Santhakumar Manicassamy, Puttur D. Prasad, Muthusamy Thangaraju. HCAR2 activation limits tumor growth by the activation of p53 signaling in breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 3304.

BackgroundCaptive cheetahs are prone to a range of unusual diseases potentially linked to unnatural diets high in muscle meat and low in collagen-rich animal fibre. In the wild, cheetahs typically eat whole prey diets not easily replicated in a captive setting. Glycine is the most abundant amino acid in collagen with a key role in several metabolic pathways such as collagen biosynthesis. Several recent studies suggest that endogenous glycine production may be limited in several species.ObjectivesUsing untargeted 1H- nuclear magnetic resonance, the metabolic changes in the urine and serum of 10 adult captive cheetahs on a glycine-supplemented diet were investigated.MethodsCheetahs were fed either a meat only (control) or glycine-supplemented meat diet (30 g glycine per 1 kg meat) for four weeks, followed by a four-week cross-over. Urine and blood samples were collected at baseline and after each intervention.ResultsA total of 151 and 60 metabolites were identified in the urine and serum, respectively. Specifically, dimethylsulphone, proline, fructose, dimethylamine, trimethylamine, pyroglutamic acid, 1,3-diaminopropane, dihydrothymine, methylmalonic acid and pimelic acid contributed to metabolome differences in the urine. In serum, glutamic acid, threonine, α-aminobutyric acid, glucose-6-phosphate, ethanolamine, methionine and propionic acid were highlighted. These metabolites play various metabolic roles in energy production, immune function, protein and collagen biosynthesis or as products of gut microbiome fermentation.ConclusionGlycine supplementation influenced threonine sparing, pyrimidine biosynthesis pathways and bacterial fermentation products, although the implications of these findings on the health of captive cheetahs is unknown. Future studies should use a targeted approach to further elaborate on these pathways.

The human gut microbiota (HGM) is a complex ecosystem subtly dependent on the interplay between hundreds of bacterial species and numerous metabolites. Dietary phenols, whether ingested (e.g., plant-derived guaiacol, mequinol, or resveratrol) or products of bacterial fermentation (e.g., p-cresol), have been attributed with influencing bacterial growth and host health. They are cleared by phase II metabolism, one form utilizing β-d-glucuronidation, but encounter bacterially derived glucuronidases capable of hydrolyzing them to release their phenolic and glucuronic acid moieties with potential effects on host cells or the surrounding bacterial population. Tools to enable the detailed study of their activity are currently lacking. Syntheses of β-d-glucuronides from methyl 1,2,3,4 tetra-acetyl β-d-glucopyranosyluronate by direct glycosylation with 2-, 3-, or 4-methoxy- and 4-fluorophenol acceptors employing trimethylsilyl triflate catalysis are reported. Yields (methoxy series) were modest. An improved route from methyl 1,2,3,4-tetra-acetyl β-d-glucopyranosyluronate via selective anomeric deprotection (N-methyl piperazine) and conversion to an α-trichloroacetimidate glycosyl donor was employed. Coupling with 2- and 3-methoxyphenol acceptors and deprotection provided 2- and 3-methoxyphenyl β-d-glucuronides in 2-fold improved overall yield. These naturally occurring methoxyphenyl glucuronides augment available model substrates of dietary glucuronides, which include 3- and 4'-linked resveratrol. The use of model glucuronides as substrates was illustrated in studies of β-d-glucuronidase activity employing cell lysates of 9 species of HGM (Bacteroidetes), revealing distinct outcomes. Contrasting effects on bacterial growth were also observed between the free phenolic components, their respective glucuronides, and glucuronic acid. The glucuronide of 4-fluorophenol provided sensitive and background-free detection of β-glucuronidase activity using 19F NMR.

The genus Senna contains globally distributed plant species of which the leaves, roots, and seeds have multiple traditional medicinal and nutritional uses. Notable chemical compounds derived from Senna spp. include sennosides and emodin which have been tested for antimicrobial effects in addition to their known laxative functions. However, studies of the effects of the combined chemical components on intact human gut microbiome communities are lacking. This study evaluated the effects of Juemingzi (Senna sp.) extract on the human gut microbiome using SIFR® (Systemic Intestinal Fermentation Research) technology. After a 48-hour human fecal incubation, we measured total bacterial cell density and fermentation products including pH, gas production and concentrations of short chain fatty acids (SCFAs). The initial and post-incubation microbial community structure and functional potential were characterized using shotgun metagenomic sequencing. Juemingzi (Senna seed) extracts displayed strong, taxon-specific anti-microbial effects as indicated by significant reductions in cell density (40%) and intra-sample community diversity. Members of the Bacteroidota were nearly eliminated over the 48-hour incubation. While generally part of a healthy gut microbiome, specific species of Bacteroides can be pathogenic. The active persistence of the members of the Enterobacteriaceae and selected Actinomycetota despite the reduction in overall cell numbers was demonstrated by increased fermentative outputs including high concentrations of gas and acetate with correspondingly reduced pH. These large-scale shifts in microbial community structure indicate the need for further evaluation of dosages and potential administration with prebiotic or synbiotic supplements. Overall, the very specific effects of these extracts may offer the potential for targeted antimicrobial uses or as a tool in the targeted remodeling of the gut microbiome.

Constipation may accelerate the progression of Parkinson's disease (PD). The quality of life in PD patients can be significantly improved when constipation is treated, hence the disease progression may be delayed. At present, the mechanism research is still at the initial stage for acupuncture in treatment of PD, focusing on neurohumoral factors, intestinal flora, bacterial fermentation products and intestinal inflammation； and there are the problems such as single intervention, thinking limitation and insufficient cooperation among disciplines. This paper systematically reviews the mechanism research progress of acupuncture for the treatment of constipation in PD so as to provide the references for the subsequent studies.

Short-chain fatty acids (SCFAs) are products of bacterial fermentation that help maintain important gut functions such as maintenance of the intestinal barrier, cell signaling, and immune homeostasis. The main SCFAs acetate, propionate, and butyrate have demonstrated beneficial effects for the host, including its importance in alleviating infections caused by pathogens such as Clostridioides difficile. Despite the potential role of SCFAs in mitigating C. difficile infection, their direct effect on C. difficile remains unclear. Through a set of in vitro experiments, we investigated how SCFAs influence C. difficile growth, sporulation, and toxin production. Similar to previous studies, we observed that butyrate decreased growth of C. difficile strain 630 in a dose-dependent manner. The presence of butyrate also increased C. difficile sporulation, with minimal increases in toxin production. RNA-Seq analysis validated our experimental results, demonstrating increased expression of sporulation-related genes in conjunction with changes in metabolic and regulatory genes, such as a putative carbon starvation protein, CstA. Collectively, these data suggest that butyrate may induce alternative C. difficile survival pathways, modifying its growth ability and virulence to persist in the gut environment. IMPORTANCE Several studies suggest that butyrate may modulate gut infections, such as reducing inflammation caused by the healthcare-associated Clostridioides difficile. While studies in both animal models and human studies correlate high levels of butyrate with reduced C. difficile burden, the direct impact of butyrate on C. difficile remains unclear. Our study demonstrates that butyrate directly influences C. difficile by increasing its sporulation and modifying its metabolism, potentially using butyrate as a biomarker to shift survival strategies in a changing gut environment. These data point to additional therapeutic approaches to combat C. difficile in a butyrate-directed manner.

Moringa oleifera is an important medicinal plant in several countries, e.g. Nigeria, USA, Turkey, Germany, Greece, and Ukraine. The abundant bioactive and nutritional properties of this plant make it useful in many and diverse areas of life including the health, cosmetic, agricultural and food industries to mention but a few. Research has found out that the presence of proteins, carbohydrates, lipids, vitamins, minerals, flavonoids, phenols, alkaloids, fatty acids, saponins, essential oils, folate, aromatic hydrocarbons, sterols, glucosinolates, glycosides among others characterise the Moringa- nutrient profile and as a result, give rise to its remedial effects on ailments such like wounds, stomach and duodenal ulcers, allergies, obesity, diabetics, inflammation, asthma, etcetera. It is the aim of this review to provide an insight into such medicinal and pharmacological remedies attributed to Moringa stating both the past and recent discoveries. This review article also takes a look into the botanical features, bioactive compounds, antinutrients, food applications, bacterial fermentation products, biosafety, industrial applications and other uses of Moringa. Finally, with belief that knowledge is progressive, we acknowledge that there are things yet undiscovered about this wonder plant that will be of value both to medicine and general life, we therefore recommend that research work continue on the plant Moringa. This article is protected by copyright. All rights reserved.

<b>Abstract ID 19075</b> <b>Poster Board 368</b> <b>Introduction:</b> Recently, evidence from both preclinical¹ and clinical² studies indicates that chronic opioid use results in intestinal dysbiosis; eliminating the dysbiotic bacteria with antibiotics prevents the onset of tolerance to the antinociceptive effects¹. However, the mechanism underlying opioid-induced dysbiosis (OID) is unclear. One mechanism by which the intestinal epithelium maintains the microbiota is through the release of antimicrobial peptides (AMPs). In the present study we tested if chronic morphine or fentanyl altered the antimicrobial activity of the intestinal epithelium. We also tested if the short-chain fatty acid, butyrate, a bacterial fermentation product known for its protective effects on the intestinal epithelium³, can prevent OID and tolerance. <b>Methods:</b> Male Swiss Webster mice (6-8 weeks) were implanted for 7-days with a morphine (75-mg) or placebo pellet and orally administered sodium butyrate (275 mg/kg) b.i.d. Tolerance to antinociception was tested on day-7 using the warm-water tail-withdrawal assay. Resected ileum tissue was incubated in DMEM/F12 for 15-18-hours to allow for the dissolution of AMPs into the media. %Antibacterial activity of the conditioned media was determined by counting the colonies of <i>E.coli</i> (Gram-negative) or <i>L.reuteri</i> (Gram-positive) on agar plates. AMPs, Regenerating islet-derived protein 3-gamma (Reg3g), active against Gram-positive bacteria, and alpha-defensin-5 (Defa5), active against Gram-positive and Gram-negative bacteria, were measured in the ileum using qRT-PCR. Experiments were also conducted in mice treated for 6 days with 0.3 mg/kg fentanyl ± butyrate (275 mg/kg) i.p. b.i.d. <b>Results:</b> %Antibacterial activity against <i>E.coli</i> or <i>L.reuteri</i> of ileum tissue supernatants from chronic morphine-treated mice was significantly reduced compared to that from placebo controls. Chronic morphine or fentanyl administration also significantly down-regulated Reg3g and Defa5 in the ileum. Butyrate prevented opioid-induced decrease in %antibacterial activity and downregulation of AMPs in the ileum. Butyrate-treated mice also did not exhibit tolerance in the warm-water tail-withdrawal assay. <b>Conclusion:</b> Chronic opioid treatment reduced AMPs in the ileum, implicating a potential mechanism underlying OID. Preventing the opioid-induced decrease in antibacterial activity with sodium butyrate precluded the development of tolerance to antinociception, thus implicating sodium butyrate as a prophylactic treatment for OID. <b>References</b> Kang M, Mischel RA, Bhave S, et&nbsp;al. The effect of gut microbiome on tolerance to morphine mediated antinociception in mice. <i>Sci Rep</i>. 2017;7(1):42658. doi:10.1038/srep42658 Cruz-Lebrón A, Johnson R, Mazahery C, et&nbsp;al. Chronic opioid use modulates human enteric microbiota and intestinal barrier integrity. <i>Gut Microbes</i>. 2021;13(1). doi:10.1080/19490976.2021.1946368 Tan J, McKenzie C, Potamitis M, Thorburn AN, Mackay CR, Macia L. The role of short-chain fatty acids in health and disease. <i>Adv Immunol</i>. 2014;121:91-119. doi:10.1016/B978-0-12-800100-4.00003-9 This work was supported by the National Institute of Health grant: P30 DA033934

Mechanisms of carbonate precipitation induced by two model bacteria

Nanometric scale size oscillations seem to be a fundamental feature of all living organisms on Earth. Their detection usually requires complex and very sensitive devices. However, some recent studies demonstrated that very simple optical microscopes and dedicated image processing software can also fulfill this task. This novel technique, termed as optical nanomotion detection (ONMD), was recently successfully used on yeast cells to conduct rapid antifungal sensitivity tests. In this study, we demonstrate that the ONMD method can monitor motile sub-cellular organelles, such as mitochondria. Here, mitochondrial isolates (from HEK 293 T and Jurkat cells) undergo predictable motility when viewed by ONMD and triggered by mitochondrial toxins, citric acid intermediates, and dietary and bacterial fermentation products (short-chain fatty acids) at various doses and durations. The technique has superior advantages compared to classical methods since it is rapid, possesses a single organelle sensitivity, and is label- and attachment-free.

The deterrent ability of Xenorhabdus nematophila and Photorhabdus laumondii compounds as a potential novel tool for Lobesia botrana (Lepidoptera: Tortricidae) management

Low expression of the intestinal metabolite butyric acid and the corresponding memory pattern regulate HDAC4 to promote apoptosis in rat hippocampal neurons

Cryptosporidium infection induced the dropping of SCFAS and dysbiosis in intestinal microbiome of Tibetan pigs.

Studies in human microbiota dysbiosis have shown that short-chain fatty acids (SCFAs) like propionate, acetate, and particularly butyrate, positively affect energy homeostasis, behavior, and inflammation. This positive effect can be demonstrated in the reduction of butyrate-producing bacteria observed in the gut microbiota of individuals with type 2 diabetes (T2DM) and other energy-associated metabolic alterations. Butyrate is the major end product of dietary fiber bacterial fermentation in the large intestine and serves as the primary energy source for colonocytes. In addition, it plays a key role in reducing glycemia and improving body weight control and insulin sensitivity. The major mechanisms involved in butyrate regulation include key signaling pathways such as AMPK, p38, HDAC inhibition, and cAMP production/signaling. Treatment strategies using butyrate aim to increase its intestine levels, bioavailability, and improvement in delivery either through direct supplementation or by increasing dietary fiber in the diet, which ultimately generates a higher production of butyrate in the gut. In the final part of this review, we present a summary of the most relevant studies currently being carried out in humans.

To obtain insights into how bacterial and fungal microbiota and fermentation products composition are affected by storage temperature for TMR silage, which can be manufactured year-round. TMR silage was stored at 10°C, 25°C, ambient temperature (AT; 20-35°C) and 40°C. Lactic acid production was delayed when stored at 10°C, and acid production stagnated after 2 weeks when stored at 40°C. The patterns of acetic acid and ethanol production were inversely related, with ethanol production promoted at 10°C and 25°C and acetic acid production promoted at AT and 40°C. The bacterial diversity was reduced in TMR silage with high lactic acid and acetic acid content, and the fungal diversity was reduced in TMR silage with high ethanol content. The intensity of lactic acid production was accounted for by the high abundance of Lactobacillus, and its stagnated production at a substantially high storage temperature was related to an increased abundance of Bacillus. The enhanced production of acetic acid or ethanol can be explained by differences in the fungal microbiota. The integrated analysis of bacterial and fungal microbiota can provide in-depth insights into the impact of storage temperature on TMR silage fermentation.

Plasma Short-Chain Fatty Acid (SCFA) Concentrations Are Related to Cognition and Adipose Tissue Mass but Not Muscle Function in Older Adults

Influenza vaccination is an effective public health measure to reduce the risk of influenza illness, particularly when the vaccine is well matched to circulating strains. Notwithstanding, the efficacy of influenza vaccination varies greatly among vaccinees due to largely unknown immunological determinants, thereby dampening population-wide protection. Here, we report that dietary fibre may play a significant role in humoral vaccine responses. We found dietary fibre intake and the abundance of fibre-fermenting intestinal bacteria to be positively correlated with humoral influenza vaccine-specific immune responses in human vaccinees, albeit without reaching statistical significance. Importantly, this correlation was largely driven by first-time vaccinees; prior influenza vaccination negatively correlated with vaccine immunogenicity. In support of these observations, dietary fibre consumption significantly enhanced humoral influenza vaccine responses in mice, where the effect was mechanistically linked to short-chain fatty acids, the bacterial fermentation product of dietary fibre. Overall, these findings may bear significant importance for emerging infectious agents, such as COVID-19, and associated de novo vaccinations.