ABSTRACT The current study aimed to assess the effect of deep-frying with four different culinary oils on the fatty acid composition and lipid nutritional quality indices (LNQIs) of the striped seabream, Lithognathus mormyrus. As a result of frying, the fat content of the fillets increased, and the moisture content decreased in all evaluated samples. The findings revealed significant differences in fatty acid composition depending on the frying oil used. The nutritional quality indices (NQI) were all within ranges of high-lipid-quality seafood. The n-3/n-6 ratio decreased significantly after frying, mainly in sunflower oil, indicating a reduction in the nutritional value of the fried fish.

In recent decades, interest in nitrated fatty acids (NO2-FAs) has grown due to their role as endogenous signaling molecules involved in health and disease. As a result, their metabolic profiling has gained increasing attention. For metabolite analysis, GC-MS/MS offers greater sensitivity and robustness than LC-MS/MS, with more reliable annotation libraries. This study investigates metabolic dysregulation in cardiovascular disease (CVD) patients by using advanced metabolomics. A novel GC-MS/MS method for profiling NO2-FAs was developed, showing improved precision using 17-BrHDA as an internal standard compared to previous HDA-based methods. It is also the first report of alkylation and silylation derivatization of 17-BrHDA, demonstrating superior GC-MS sensitivity for pentafluorobenzyl-alkylated fatty acids over their silylated counterparts in positive ion mode. Untargeted metabolomics was applied to plasma samples from acute myocardial infarction (AMI) patients and healthy controls using both derivatization techniques. Multivariate analysis (PCA and PLS-DA) revealed distinct metabolic profiles. Key metabolites, identified based on VIP scores, were annotated via the Human Metabolome Database and literature. Findings highlight the complementary nature of both derivatization approaches for comprehensive plasma metabolome analysis. Notably, NO2-OA levels were significantly elevated (p < 0.01) in AMI patients, indicating its possibility to be utilized as a cardiovascular biomarker. This study represents the first use of alkylation derivatization in untargeted metabolomics for AMI and introduces a highly sensitive GC-MS/MS method with an innovative internal standard and optimized derivatization for cardiovascular biomarker discovery. The method demonstrates the potential to discriminate between groups of patients and healthy subjects.

Cadmium (Cd) and lead (Pb) ions are highly toxic elements present in the water, soil and sediments of the Yucatan Peninsula. The use of Cd- and Pb-resistant microorganisms as natural biosorbents could be considered an innovative strategy for the bioremediation of ecosystems contaminated with these ions. In this investigation, halophilic bacteria of the genus Brachyobacterium were identified that were tolerant to high concentrations of metal ions isolated from the coasts of Isla Arena, Mexico. Sediment parameters showed pH values ​​ > 7.6 and < 8.5; temperatures > 30°C and < 33°C; salinity > 2.0% and < 4.2%; conductivity > 2411µs/cm and < 8240µs/cm; and total solids > 1204ppm and < 4193ppm. Isolates S1p and S1a were genetically identified as Brachybacterium paraconglomeratum and Brachybacterium saurashtrense, both with 99.7% identity, according to the software employed. The minimum inhibitory concentration (MIC) values ​​indicated a tolerance of 1656mg/L of Pb for both strains; while for Cd, the tolerance values ​​were 591mg/L and 236mg/L for S1p and S1a, respectively. Additionally, FT-IR analysis demonstrated that, most likely the functional groups involved in this metal-bacteria interaction are OH-, NH-, and/or COOH-, associated with proteins, lipids and fatty acids in cell walls of bacteria, as also reported by other authors. In this study, we observed that, at a pH of 6.5 and a time of 48h, a maximum biosorption capacity of 58mg/L was obtained. This work presents the biosorption capacity of cadmium and leads ions from halophilic bacteria of the genus Brachybacterium isolated from undisturbed sites and opens the possibility of exploring this methodology in other scenarios.

Abstract Aims Trichoderma is a filamentous fungus beneficial to crops and widely used as biofungicide. Its mechanisms of action as a biological control agent against phytopathogenic fungi include mycoparasitism and the induction of plant defense responses. On the other hand, Sclerotinia sclerotiorum is an important pathogen for Brassica crops, effectively suppressed by Trichoderma . The aim of this work is to determine whether T. hamatum can release elicitors from the cell wall of S. sclerotiorum that activate plant systemic defenses against the pathogen. Methods Liquid fermentation of T. hamatum on S. sclerotiorum mycelium was performed. In the resulting fungal filtrates, chitinase and β-endoglucanase activities were quantified, along with the amounts of glucosamine and glucan oligomers produced. These filtrates were subsequently applied to the roots of broccoli plants ( Brassica oleracea var. italica ), which were later foliar-infected with the pathogen. Lesions produced were measured and different systemic defensive responses were evaluated through hormonomics, glucosinolate profiling and non-targeted metabolomics. Results In fungal filtrates of T. hamatum cultured on S. sclerotiorum , chitinase (7.56–8.32 units/mL) and β-endoglucanase (3.45 units/mL) activity was determined. These filtrates also contained the highest amounts of glucosamine (0.75 g/L) and glucan oligomers (43.8 g/L). When applied to broccoli plants, the filtrates triggered a systemic defense response that was effective against the pathogen. This response was mediated by the hormones jasmonic acid, isopentenyladenine and ethylene, leading to the accumulation of antifungal compounds in the leaves, including glucobrassicin, niacin and several fatty acids. This defensive induction was not observed with glucosamine oligomers. Conclusions Therefore, T. hamatum releases glucan oligomers from the cell wall of S. sclerotionum which may act as potential elicitors of systemic plant defenses. Highlights • T. hamatum releases oligomers from the S. sclerotiorum cell wall by enzymatic action. • Glucan oligomers, but not glucosamine oligomers, induce systemic plant resistance. • Systemic resistance is mediated by ethylene/jasmonic acid, a response to necrotrophs. • Reduced S. sclerotiorum -foliar infection is due to an accumulation of glucobrassicin and niacin. • Trichoderma is able to release plant defense elicitors from the pathogen cell wall.

The limited efficacy of programmed death-ligand-1 (PD-L1) monoclonal antibodies (aPD-L1) in triple-negative breast cancer (TNBC) is largely attributable to the immunosuppressive tumor microenvironment (TME). Notably, the abundance of cancer-associated adipocytes (CAAs) constitutes a distinctive feature of the TNBC microenvironment, contributing significantly to its immunosuppressive nature. CAAs upregulate cluster of differentiation 36 (CD36), a fatty acid translocase on tumor cells, thereby promoting excessive fatty acids (FAs) uptake and lipid droplet (LD) accumulation, which starve immune cells and reinforce immunosuppression through this metabolic adaptation. Berberine (BBR), a bioactive alkaloid derived from Rhizoma coptidis, has previously been shown to ameliorate lipid metabolism disorders through downregulation of CD36 in metabolic diseases such as hepatic steatosis. We therefore hypothesize that BBR inhibits CD36-mediated FAs uptake and reduces LD accumulation in tumor cells, representing a novel mechanism that remains unexplored in the context of TNBC. In this study, we demonstrated that BBR counteracts the tumor-promoting effects of CAAs in 4T1 cells by inhibiting CD36 upregulation and its mediated FAs uptake, thereby reducing CAA-induced LD accumulation and ultimately suppressing tumor cell proliferation. Furthermore, BBR remodeled the TME by enhancing CD8+ T cell recruitment and activity, while reducing immunosuppressive factors. In order to improve the sustained release of BBR at the tumor site and overcome its poor aqueous solubility, we created a thermosensitive hydrogel-based nanoparticle system (BBR-NPs-GEL). This injectable hydrogel demonstrated favorable thermosensitive gelation and shear-thinning behavior, making it suitable for localized administration. It exhibited a gelation temperature of 35.3 ± 0.2 °C and a sustained release profile with 52% of BBR released within 48 h. In 4T1Fluc tumor-bearing mice, BBR-NPs-GEL significantly suppressed tumor growth and remodeled the TME, as evidenced by increased infiltration of CD8+ T cells (+8.49%), activation of dendritic cells (+8.39%), and a shift toward M1-macrophages (+39.9%), accompanied by a reduction in M2-macrophages (-19.13%). Importantly, when combined with aPD-L1 therapy, the treatment elicited synergistic antitumor effects, resulting in enhanced tumor regression. This combination strategy effectively overcame metabolic immunosuppression and reversed immune resistance in TNBC.

Clear cell renal cell carcinoma (ccRCC) is characterized by disrupted lipid metabolism, traditionally attributed to VHL mutations and HIF stabilization. Here, we identified CEBPB as an epigenetically upregulated, VHL-independent transcription factor driving ccRCC tumorigenesis. CEBPB was regulated by H3K27ac and H3K4me and transcriptionally repressed the tumor-suppressive glycerol-3-phosphate dehydrogenase 1-like protein (GPD1L), thereby elevating dihydroxyacetone phosphate (DHAP)-derived ether lipid synthesis and enhancing Akt signaling. This activation suppressed CPT1A expression, inhibiting fatty acid oxidation (FAO) and leading to lipid accumulation, as found by lipidomics and isotope tracing. Loss of CEBPB reduced ether lipids, reactivated CPT1A, and impaired Akt signaling, diminishing tumor growth and lipid content in vitro and in vivo. Restoration of ether lipids or Akt activity rescued these effects. Importantly, CEBPB expression and enhancer activation were not modulated by VHL status and it could be targeted pharmacologically. The CEBPB-GPD1L-ether lipid-Akt-CPT1A axis is proposed as a new druggable driver in ccRCC integrating epigenetics, transcription, intermediary metabolism and oncogenic signaling.

Introduction Microbial additives can improve silage quality in lowland areas. However, Saccharomyces cerevisiae and Lactic Acid Bacteriacan efficacy on whole-plant maize silage under Tibet’s hypoxic and cold environment, have not been explored. Methods In this experiment, whole corn plants cultivated in Dazi District, Lhasa City, Xizang (Tibet) Autonomous Region, were selected as silage raw materials. The treatment group was added 0.5 kg of microbial additives per ton of silage. The addition levels for both Saccharomyces cerevisiae and Lactic Acid Bacteria were ≥ 1 × 107 CFU·g-1 FM). The quality of silage and its in vitro fermentation characteristics were determined on 0, 30 and 60 days of fermentation, respectively. Subsequently, dairy cows were fed with silage after 60 days of fermentation to evaluate milk production and milk quality. Results The results indicated that the lactic acid content in the treatment group was increased significantly on 30 and 60 days of fermentation ( p &amp;lt; 0.05). In addition to Simpson’s index, alpha diversity was significantly affected by the fermentation day × treatment interaction ( p &amp;lt; 0.05). At 60 days of fermentation, the abundance of Firmicutes phylum in the treatment group was significantly higher than that in the control group ( p &amp;lt; 0.05). The abundance of genera such as Acetobacter and Latilactobacillus was significantly decreased ( p &amp;lt; 0.05), while the abundance of the genus Weissella was significantly increased ( p &amp;lt; 0.05). Dairy cows were fed 60-day maize silage, the milk protein content and total solid content in the treatment group were significantly higher than that in the control group ( p &amp;lt; 0.05). The levels of dry matter degradation rate, ammonia nitrogen and total volatile fatty acids in the in vitro fermentation of maize silage in the treatment group on the 60th day of fermentation were significantly higher than that in the control group ( p &amp;lt; 0.05). Conclusion In Xizang (Lhasa, China), the addition of microbial additives has significantly improved the quality and nutritional value of whole corn silage plants and enhanced the milk quality of local dairy cows. This provides a theoretical basis for the application of microbial additives from the Qinghai-Tibet Plateau to agricultural crops.

The utilization of insect protein, specifically black soldier fly larvae meal (BSFLM), offers a viable alternative, sustainable, and nutrient-rich ingredient for pet foods. However, limited information is available on optimal inclusion levels and its functional properties. The objective of the study was to evaluate the effects of incremental levels of BSFLM as a partial or full substitute for chicken by-product meal (CM) in extruded diets. Thirty healthy adult Beagles were assigned to one of three dietary treatments: a control diet with CM, a diet containing 15% BSFLM, and a diet with 30% BSFLM, where CM was partially or fully replaced. Following a 14-day adaptation period on the control diet, the dogs were fed the test diets for 56 days. No significant differences were observed in food intake, fecal output, or apparent total tract digestibility (ATTD) of dry matter, crude protein, and fat across the treatment groups (P > 0.05). However, dogs fed the 30% BSFLM diet showed a reduction in ATTD of organic matter (81.2%) and total dietary fiber (18.9%) compared to the control group (83.2% and 30.3%, respectively; P < 0.05), likely due to the chitin content acting as insoluble fiber. Fecal microbiota analysis showed no changes in alpha diversity; however, unweighted UniFrac analysis revealed significant shifts in beta diversity associated with diet and time. The inclusion of BSFLM-modulated fecal metabolites resulted in lower concentrations of short-chain fatty acids and phenols in the 30% BSFLM group (P < 0.05). Serum biomarkers of oxidative stress and inflammation, including a range of cytokines and antioxidant enzymes, were unaffected by the dietary treatments, indicating no adverse health effects. These findings support the use of BSFLM as a viable and environmentally friendly protein ingredient in canine nutrition, offering a promising alternative to traditional protein sources while contributing to the sustainability of pet food production.

Most omega-3 supplements contain eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in defined ratios, yet their pharmacokinetic and anti-inflammatory effects are not fully understood. This systematic review and meta-analysis evaluated how EPA:DHA dosing ratios, alongside EPA+DHA daily dose, influence blood fatty acid profiles and inflammatory markers across 96 clinical trials published before February 2025. Standardized mean differences with 95% confidence intervals were calculated. EPA+DHA supplementation significantly increased EPA:DHA blood ratios and blood EPA+DHA levels, with larger effects in healthy individuals. Supplementation also reduced arachidonic acid (AA) and key inflammatory markers, C-reactive protein (CRP), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6), particularly in participants with underlying health conditions. Ratios <1.0 produced the greatest cytokine reductions, whereas ratios ≥1.0 most effectively increased the EPA:DHA blood ratio and lowered AA. EPA+DHA doses of 1-3 g/day were associated with the most consistent reductions in CRP, TNF-α, and IL-6. Linear regression showed a strong association between the EPA:DHA dosing ratio and the EPA:DHA blood ratio and identified the EPA:DHA blood ratio as the strongest predictor of AA reduction. These findings indicate distinct ratio- and dose-dependent effects of EPA and DHA and highlight the importance of optimizing omega-3 formulations to enhance bioavailability and anti-inflammatory outcomes.

In most animals, excess dietary energy is stored as lipids in specialized tissues, such as the liver in vertebrates or the hepatopancreas and fat body in invertebrates, which function as energy reservoirs for reproduction. In cephalopods, however, dietary energy is rapidly mobilized from the digestive gland for growth rather than stored for reproduction. How excess energy is allocated for reproduction activity in cephalopods remains largely unclear. Lipogenesis is initiated by acetyl-CoA carboxylase (ACC), which converts acetyl-CoA derived from dietary carbon sources into malonyl-CoA; subsequent synthesis of saturated fatty acids is catalyzed by fatty acid synthase (FAS). Using the bigfin reef squid as a model, we investigate the role of fas in female development. fas mRNA was highly expressed in ovaries but weak in other tissues, including the lipid-rich digestive gland. fas showed female-biased expression in gonads, with level highest in juvenile ovaries and progressively decreasing to their lowest in mature ovaries. Expression was also high in primary and multiple follicular oocytes but declined in later stages. In situ hybridization and immunohistochemistry confirmed fas mRNA and protein localization in oocytes, particularly in primary and multiple follicular oocytes. In vitro ovarian culture further showed that inhibiting FAS activity enhanced somatic cell proliferation. Together, these findings suggest that squid ovary is a primary site of fatty acid synthesis, supporting early oocyte growth and membrane biogenesis in the absence of dedicated lipid storage tissues. The decline of FAS activity during oogenesis, and the associated reproduction in fatty acid synthesis, may act as a regulatory signal to promote somatic cell proliferation.

Transportation is a major stressor in livestock production, adversely affecting animal welfare, physiological status, and meat quality. Oxidative stress and metabolic imbalances induced by transport conditions can lead to significant economic losses. The use of natural antioxidants has been proposed as a strategy to mitigate these negative effects. This study aimed to evaluate the effects associated with intramuscularly administered quercetin (QUE) and grape seed extract (GSE) prior to transport during the winter season on body weight, serum biochemical responses, oxidative stress markers, and meat quality parameters, including carcass pH and fatty acid profile in sheep subjected to road transport. Twenty-four 12-month-old Akkaraman sheep were randomly allocated to four groups (n = 6): control, GSE50 (50 mg kg-1 grape seed extract), GSE100 (100 mg kg-1 grape seed extract), and QUE100 (100 mg kg-1 quercetin). Malondialdehyde (MDA) levels differed among groups, with lower values observed in antioxidant-treated animals; however, baseline variability may have contributed to post-transport differences. Transport led to significant body weight loss in all groups (P < 0.001), with the least reduction observed in the QUE100 group (P = 0.105 among groups). In contrast, carcass pH values showed minimal variation among groups, with differences not exceeding 0.22 units (P = 0.155), and fatty acid composition of longissimus thoracis (LT) muscle (P > 0.05 for all fatty acids) did not differ significantly among groups. These findings suggest that natural bioactive compounds may be associated with improved physiological responses under transport-related stress conditions. © 2026 Society of Chemical Industry.

Salinity stress represents a critical environmental constraint that significantly limits the development of tilapia aquaculture in brackish water environments. Its substantial impacts on fundamental physiological processes in fish, particularly osmotic balance, energy metabolism, and antioxidant defense mechanisms, have become a major scientific concern in aquaculture research. To systematically elucidate the molecular mechanisms underlying the response of genetically improved farmed tilapia (Oreochromis niloticus) to salinity stress and to test the hypothesis that it adapts through metabolic reprogramming for energy reallocation under such conditions, this study employed an integrated transcriptomic and metabolomic approach. Through a rigorously controlled experimental design with freshwater (0‰) as the control group and brackish water (24‰) as the experimental group, we conducted a comprehensive analysis of dynamic changes in gene expression profiles and metabolite spectra in the liver tissues of experimental fish. The study yielded the following key findings: First, salinity stress significantly suppressed growth performance indicators, including body weight and length, while simultaneously inducing extensive transcriptomic restructuring and profound metabolic remodeling in liver tissue. A total of 1529 differentially expressed genes (including 399 up-regulated and 1130 down-regulated genes) and 127 significantly differential metabolites were identified. Second, the organism achieved strategic reallocation of energy resources through coordinated suppression of multiple energy-consuming anabolic pathways, particularly steroid biosynthesis and fatty acid metabolism, with the remarkable down-regulation of Fasn, a key gene in the fatty acid synthesis pathway, being especially prominent. Energy-sensing and metabolic homeostasis regulatory networks played a central coordinating role in this process, guiding the organism through metabolic reprogramming by regulating downstream metabolic nodes. From a multi-omics integrative perspective, this study provides in-depth insights into the sophisticated metabolic remodeling and energy allocation strategies employed by GIFT to cope with salinity stress. These findings, particularly the suppression of fatty acid biosynthesis and the reprogramming of glycolysis/gluconeogenesis pathways, not only elucidate the molecular mechanisms by which teleosts achieve environmental adaptation through energy reallocation, but also provide actionable molecular targets for the selective breeding of salinity-resilient tilapia strains.

Avian infectious bronchitis virus (IBV) belongs to the genus Gammacoronavirus (family Coronaviridae), causes severe multi-system disease in chickens, inflicting major global economic losses. The molecular interplay between IBV and host metabolic networks remains poorly understood. Through integrated transcriptomic, metabolomic, and lipidomic profiling of oviduct tissues from specific-pathogen-free (SPF) chickens infected with the IBV QXL strain, we demonstrate tripartite metabolic reprogramming: (1) redirected glucose flux through the pentose phosphate pathway (PPP) to fuel nucleotide synthesis, (2) rewired lipid metabolism to prioritize de novo membrane biogenesis over fatty acid β-oxidation, and (3) orchestrated glycerophospholipid remodeling. This integrated analysis revealed a coordinated upregulation of fatty-acid biosynthesis genes and accumulation of specific glycerophospholipids and eicosanoids. Mechanistically, IBV co-opts the Warburg effect and PPP activation while uniquely suppressing fatty acid β-oxidation to channel fatty acids toward lipid droplets (LDs) biogenesis. Phosphatidylserine (PS) overproduction (e.g. 2.55-fold increase in PS(22:0/22:6)) and phospholipase A2 (PLA2)-mediated lysophospholipids (Lyso-PLs) and eicosanoids generation (e.g. 7.09-fold increase in prostaglandin E2 (PGE2)) emerged as critical regulators of membrane dynamics and inflammatory signaling. This process was centrally coordinated by the significant activation of peroxisome proliferator-activated receptor (PPAR) (e.g. 1.74-fold increase in ACSL1) and transforming growth factor-beta (TGF-β) (e.g. significant increase in p-SMAD2) signaling pathways, directly linking lipid remodeling to immunomodulation. Functionally, targeting acetyl-CoA carboxylase (ACC) or glucose-6-phosphate dehydrogenase (G6PD), alongside TGF-β pathway modulation, synergistically curtailed viral replication in vitro. Our findings delineate a critical PPAR-TGF-β cross-talk that governs lipid remodeling during infection and identify host metabolic nodes that are potentially targetable for antiviral intervention.

Ambient air pollution is associated with heart failure (HF), but underlying biological mechanisms remain unclear. We aimed to elucidate metabolic pathways linking air pollution exposure with HF. This prospective cohort study analysed 229 812 UK Biobank participants with nuclear magnetic resonance metabolomics data. Air pollution score was constructed by fine particulate matter, coarse particulate matter, nitrogen dioxide and nitrogen oxides. Air pollution-associated metabolic signatures were identified using elastic net regression among 251 circulating metabolites. Cox regression evaluated associations between metabolic signatures and incident HF risk. Mediation analysis quantified metabolic signatures' role in air pollution-HF relationships. During median 13.1-year follow-up, 8986 participants (3.9%) developed HF. We identified 53 metabolic metabolites reflecting air pollution exposure, comprising lipoprotein metabolism markers (22.6%), fatty acids (17.0%) and amino acids (13.2%), which were used to construct the air pollution-related metabolic signatures score. After adjustment for confounding factors, each SD increase in the metabolic signatures was associated with 8% elevated HF risk (HR 1.08, 95% CI 1.06 to 1.11). Participants in the highest quantile showed a 24% increased HF risk compared with those in the lowest quantile (HR 1.24, 95% CI 1.16 to 1.3). The metabolic signatures mediated 13.08% (95% CI 12.15% to 15.71%) of air pollution-HF associations, with lipoprotein metabolism and fatty acid signatures as primary mediators. Air pollution was associated with increased HF risk, with metabolic perturbations appearing to play a mediating role. These metabolic signatures provide insights into potential mechanisms linking air pollution to cardiovascular outcomes.

Lipin, a phosphatidic acid phosphatase, catalyzes the conversion of phosphatidic acid to diacylglycerol, a key step in triacylglycerol (TAG) biosynthesis, and acts as a transcriptional coregulator of lipid metabolism. In this study, the Lipin-encoding gene (McLipin) was overexpressed in Mucor circinelloides WJ11 to evaluate its impact on lipid accumulation under submerged (SmF) and solid-state fermentation (SSF). Under SSF, McLipin overexpression increased cell dry weight by 19% in spores and 36.9% in mycelia compared to the control strain. Total fatty acid (TFA) content increased by 35.7% in mycelia, while in spores it increased from 10% to 18% of dry cell weight, indicating a substantial enhancement in lipid yield. Fatty acid profiling showed elevated proportions of C16:0, C18:2, and C18:3 and a reduced C18:1 content. RT-qPCR indicated that McLipin expression peaked at 24 h under SSF, coinciding with upregulation of ACC, FAS, cME, and NADPH-generating genes. These results suggest that McLipin overexpression reprograms lipid metabolism, significantly enhancing TAG biosynthesis under SSF for potential industrial lipid production.

FAALs (fatty acyl‐AMP ligases) recruit and incorporate fatty acids during the biosynthesis of secondary metabolites. Their diversity, distribution, and substrate specificity remain poorly understood, which limits functional predictions from sequence data. In this study, we explored the prevalence and diversity of FAAL enzymes across the tree of life and show that these are widely distributed in secondary metabolite‐rich bacteria and predominantly in the context of polyketide and non‐ribosomal peptide biosynthetic pathways. FAALs were also found to be present in certain archaea and eukaryotic groups. The phylogenetic placement of FAALs was not correlated to the chain length of the fatty acids that they activate and load. Therefore, we developed a bioinformatics and AI workflow (FAALPred) to predict the chain length of the fatty‐acid substrate of a given FAAL sequence. The robustness and accuracy of the predictions generated by FAALPred were validated using independent in vitro and in silico data. We anticipate that FAALPred will not only accelerate secondary metabolite structural predictions and subsequent discovery from FAAL‐associated pathways but also facilitate engineering of lipoylation. FAALPred is available at https://faalpred.ciimar.up.pt/.

Metabolic dysfunction-associated steatohepatitis related hepatocellular carcinoma (MASH-HCC) is a distinct HCC subtype characterized by lipid accumulation, impaired fatty acid oxidation (FAO), immune evasion, and resistance to immunotherapy. In this study, we observed elevated levels of L-carnitine-a classical FAO activator-and its transporter OCTN2 in MASH-HCC. Mechanistically, L-carnitine is redirected from FAO promotion to buffering intracellular acetyl groups via conversion to acetyl-L-carnitine, leading to acetyl group depletion. This disrupts protein acetylation through two distinct pathways: reduced acetylation of p53 weakens its tumor-suppressive signaling and promotes tumor progression, while decreased acetylation of histone H3 impairs MHC-I antigen presentation, facilitating immune evasion. We further identified that the lncRNA LINCMD1 competitively bound the E3 ligase DZIP3, sequestering it in the nucleus and preventing its interaction with cytoplasmic OCTN2. This inhibited K48-linked ubiquitination of OCTN2 and stabilized its protein expression, further amplifying L-carnitine accumulation. To therapeutically target this axis, we developed a liver-specific lipid nanoparticle (LNP)-delivered antisense oligonucleotide against the DZIP3-binding region of LINCMD1, which restored p53 and MHC-I pathways and enhanced anti-PD-1 efficacy in vivo. Together, our findings uncover a noncanonical carnitine-driven metabolic-epigenetic-immune bypass in MASH-HCC and identify the LINCMD1/DZIP3/OCTN2-L-carnitine axis as a potential therapeutic target.

Introduction Asthma is a multifactorial disease influenced by genetic and environmental factors, including diet. The gut microbiome contributes to airway inflammation via the gut-lung axis, partly through production of short chain fatty acids (SCFAs) from bacterial fermentation of dietary fiber. We hypothesized that dietary fiber supplementation could modulate the gut microbiome and increase SCFAs in children with asthma. Methods This is a double-blind, placebo-controlled trial of children who were randomized to consume 12 g of soluble corn fiber (SCF) as a supplement to their usual daily diet (50% the recommended daily fiber intake) or placebo for 4–6 weeks ( clinicaltrials.gov NCT03673618). Dietary surveys, asthma symptom questionnaires, fecal, blood and nasal samples were collected before and after the intervention period to quantify fiber intake, asthma control, nasal and gut microbiome, and serum short chain fatty acids (SCFAs). Results Of the 20 children enrolled, 15 completed the intervention with an average adherence rate of 83%. SCFA concentrations and gut microbiome changes varied by individual and treatment group. No significant differences in gut or nasal alpha or beta diversity were observed between groups post-intervention. However, differential abundance analysis showed a trend toward increased Bifidobacterium in the SCF group compared to placebo (ANCOM-BC p = 0.0004, FDR q = 0.073). Discussion Supplementation of 50% of recommended daily fiber intake had minimal impact on asthma symptoms, the microbiome, or SCFA levels. Future studies should consider higher fiber doses, different fiber types, or targeting individuals with low baseline fiber intake to account for observed variability in microbiome and SCFA responses. Clinical Trial Registration https://clinicaltrials.gov/study/NCT03673618 , identifier NCT03673618.

To investigate gut microbial alteration and their functional consequences in obesity (OB)-related knee osteoarthritis (OA) by integrating microbiome with metabolomic, proteomic, and dietary data. Fecal and fasting plasma samples were collected from 91 knee OA patients and 12 OA-free controls, classified into four subgroups based on OB and OA status: 66 OB+OA+, 25 OB-OA+, 5 OB+OA-, and 7 OB-OA-. 16S rRNA gene sequencing was performed to profile gut microbiota. MaAsLin2 modelling was applied, and dietary intake was incorporated into the models. Plasma metabolomics (n=630 metabolites) and proteomics (n=5,416 proteins) were integrated with microbial signatures to assess functional associations. OB+OA+ patients exhibited significantly lower a- and β-diversity than OB-OA+ (p<0.05). Seventeen microbial taxa were identified to be significantly associated with OB+OA+ (all p<7.65×10-5 after correcting tests for 654 ASVs), and 16 of them remained significant after adjustment for age, sex, antibiotic use, and dietary intake. PICRUSt2-based predictive analysis on these taxa suggested that bile acid biosynthesis was upregulated in OB+OA+ group. These taxa were correlated with 376 metabolites (p<0.05) with enrichment in fatty acid biosynthesis, linoleic/arachidonic acid metabolism, and propanoate metabolism pathways. They were also associated with 146 proteins (p<0.001) with enrichment in PI3K-Akt signalling, ECM-receptor interaction, and lipid/atherosclerosis pathways. OB+OA+ patients exhibited significant gut microbial dysbiosis associated with systemic metabolic and proteomic alterations relevant to OA pathophysiology. The microbiome-metabolome-proteome axis may provide mechanistic insights into worsened OA outcomes in OB individuals and could inform microbiome-targeted interventions.

Feather follicles are specialized skin appendages that are essential for thermoregulation, protection, and down production in birds, forming through complex genetic and epigenetic interactions during embryogenesis. In this study, we examined skin and follicle development in Hungarian white goose embryos, focusing on dynamic epigenetic-transcriptomic changes. Histology showed smooth epidermis at E10, feather buds at E13, and columnar follicles with medullary tissue and secondary follicles at E18. Transcriptomics revealed 1327 and 1847 DEGs enriched in epidermal development, differentiation, and adhesion. Primordial initiation at E10-E13 featured Wnt, TGF-β, and melanogenesis, whereas follicle formation at E13-E18 involved lipid metabolism and VEGF signaling. Keratinization genes were continuously upregulated, and Wnt, Shh, and muscle pathways were activated late. Key regulators included LEF1, MSX2, and FOXN1. ATAC-seq showed dynamic chromatin accessibility, stage-specific promoter openness, and motifs for YY1, KLF5, and KLF4. Differentially accessible regions enriched genes in Wnt and TGF-β signaling, cell adhesion, and mitophagy, shifting from proliferation and basic metabolism at E10-E13 to differentiation, lipid metabolism, and homeostasis at E13-E18. Integrated analyses linked fatty acid metabolism, MAPK, and FoxO signaling to differentiation, while downregulation of redox and migration pathways preserved homeostasis. Some fatty acid metabolism and cell polarity genes increased expression despite reduced accessibility at E13-E18, indicating epigenetic pre-programming and post-transcriptional interplay. This work delineates coordinated epigenetic-transcriptional regulation of goose embryonic skin and feather follicle morphogenesis, offering insights for avian and vertebrate skin appendage studies.