Non-targeted Metabolomics Analysis Research Articles

Regulations of Citrus Pectin Oligosaccharide on Cholesterol Metabolism: Insights from Integrative Analysis of Gut Microbiota and Metabolites

(1) Background: Recently, academic studies are demonstrating that the cholesterol-lowering effects of pectin oligosaccharides (POSs) are correlated to intestinal flora. However, the mechanisms of POS on cholesterol metabolisms are limited, and the observations of intestinal flora are lacking integrative analyses. (2) Aim and methods: To reveal the regulatory mechanisms of POS on cholesterol metabolism via an integrative analysis of the gut microbiota, the changes in gut microbiota structure and metabolite composition after POS addition were investigated using Illumina MiSeq sequencing and non-targeted metabolomics through in vitro gut microbiota fermentation. (3) Results: The composition of fecal gut flora was adjusted positively by POS. POS increased the abundances of the cholesterol-related bacterial groups Bacteroidetes, Bifidobacterium and Lactobacillus, while it decreased conditional pathogenic Escherichia coli and Enterococcus, showing good prebiotic activities. POS changed the composition of gut microbiota fermentation metabolites (P24), causing significant changes in 221 species of fermentation metabolites in a non-targeted metabolomics analysis and promoting the production of short-chain fatty acids. The abundances of four types of cholesterol metabolism-related metabolites (adenosine monophosphate, cyclic adenosine monophosphate, guanosine and butyrate) were significantly higher in the P24 group than those in the control group without POS addition. (4) Conclusion: The abovementioned results may explain the hypocholesterolemic effects of POS and promotion effects on cholesterol efflux of P24. These findings indicated that the potential regulatory mechanisms of citrus POS on cholesterol metabolism are modulated by cholesterol-related gut microbiota and specific metabolites.

Bifidobacterium bifidum Ameliorates DSS-Induced Colitis in Mice by Regulating Microbial Metabolome and Targeting Gut Microbiota.

Inflammatory bowel disease (IBD) is a recurrent inflammatory condition affecting the gastrointestinal tract, and its clinical treatment remains suboptimal. Probiotics have shown effectiveness in alleviating dextran sulfate sodium salt (DSS)-induced colitis, exhibiting strain-specific anti-inflammatory properties. In this study, we compared the therapeutic effects of five strains of Bifidobacterium bifidum isolated from healthy adult feces on DSS-induced colitis in mice. Additionally, we investigated the underlying mechanisms by examining gut microbiota composition and microbial metabolome. Our findings highlighted the superior efficacy of B. bifidum M1-3 compared to other strains. It significantly improved colitis symptoms, mitigated gut barrier disruption, and reduced colonic inflammation in DSS-treated mice. Moreover, gut microbiota composition analysis revealed that B. bifidum M1-3 treatment increased the abundance and diversity of gut microbiota. Specifically, it significantly increased the abundance of Muribaculaceae, Lactobacillus, Bacteroides, and Enterorhabdus, while decreasing the abundance of Escherichia-Shigella. Furthermore, our nontargeted metabolomics analysis illustrated that B. bifidum M1-3 treatment had a regulatory effect on various metabolic pathways, including tyrosine metabolism, lysine degradation, and tryptophan metabolism. Importantly, we confirmed that the therapeutic efficiency of B. bifidum M1-3 was dependent on the gut microbiota. These results are conducive to the development of probiotic products for alleviating colitis.

Impact of polyvinyl chloride microplastic exposure on the hepatopancreas metabolism of Pinctada fucata martensii: Insights from metabolomics

Microplastics (MPs) are prevalent throughout the oceans and pose a significant global environmental concern due to their detrimental effects on various living organisms. While researchers have investigated the toxicity mechanisms of MPs in marine species, there is a need for a more comprehensive understanding, particularly regarding the potential adverse effects on the growth and metabolism of bivalves. In this study, indoor exposure experiments were conducted on Pinctada fucata martensii to examine the impact of commercial polyvinyl chloride (PVC)-MPs (50 μm, 15 mg/L). The aim was to determine the effects of MPs on the hepatopancreas metabolism of P. f. martensii. Non-targeted metabolomics analysis indicated alterations in the hepatopancreatic metabolic of P. f. martensii following exposure to PVC-MPs. A total of 32, 27, and 154 differential metabolites (SDM) were identified in EG (1-d) vs CG (1-d), EG (15-d) vs CG (15-d), and EG (1-d) vs EG (15-d), respectively (VIP >1 and P < 0.05). Further examination of metabolic pathways revealed that PVC-MPs disrupted the metabolism of amino acids, particularly alanine, aspartic acid, and glutamic acid metabolism; this suggests that the organism may have developed neurotoxicity. Sphingolipid metabolites showed a significant up-regulation in both EG (15-d) vs CG (15-d), as well as in EG (1-d) vs EG (15-d), indicating lipid peroxidation in the hepatopancreas of P. f. martensii following PVC-MPs exposure. Additionally, in EG (1-d) vs CG (1-d), starch and sucrose metabolism were significantly inhibited, glycolysis or gluconeogenesis pathways were promoted, and changes in energy metabolism were promoted. This study sheds light on the impact of MPs on the metabolism of bivalves, offering fresh insights into the detrimental effects of MPs on aquatic organisms.

Inhibition mechanism of Rhizoctonia solani by pectin-coated iron metal-organic framework nanoparticles and evidence of an induced defense response in rice

Nanocrop protectants have attracted much attention as sustainable platforms for controlling pests and diseases and improving crop nutrition. Here, we reported the fungicidal activity and disease inhibition potential of pectin-coated metal-iron organic framework nanoparticles (Fe-MOF-PT NPs) against rice stripe blight (RSB). An in vitro bacterial inhibition assay showed that Fe-MOF-PT NPs (80 mg/L) significantly inhibited mycelial growth and nucleus formation. The Fe-MOF-PT NPs adsorbed to the surface of mycelia and induced toxicity by disrupting cell membranes, mitochondria, and DNA. The results of a nontargeted metabolomics analysis showed that the metabolites of amino acids and their metabolites, heterocyclic compounds, fatty acids, and nucleotides and their metabolites were significantly downregulated after treatment with 80 mg/L NPs. The difference in metabolite abundance between the CK and Fe-MOF-PT NPs (80 mg/L) treatment groups was mainly related to nucleotide metabolism, pyrimidine metabolism, purine metabolism, fatty acid metabolism, and amino acid metabolism. The results of the greenhouse experiment showed that Fe-MOF-PT NPs improved rice resistance to R. solani by inhibiting mycelial invasion, enhancing antioxidant enzyme activities, activating the jasmonic acid signaling pathway, and enhancing photosynthesis. These findings indicate the great potential of Fe-MOF-PT NPs as a new RSB disease management strategy and provide new insights into plant fungal disease management.

Nicotinamide riboside ameliorates survival time and motor dysfunction in an MPTP-Induced Parkinson's disease zebrafish model through effects on glucose metabolism and endoplasmic reticulum stress

Nicotinamide riboside (NR) is a precursor and exogenous supplement of nicotinamide adenine dinucleotide (NAD+). NR has been shown to play a beneficial role in a variety of neurodegenerative diseases. A phase 1 clinical trial identified NR as a potential neuroprotective therapy for Parkinson's disease (PD). However, the mechanism of action of NR in PD has not been fully elucidated. Therefore, the present study aimed to investigate the potential effects of NR on a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced PD model in zebrafish and its underlying mechanisms. The results showed that NR improved motor dysfunction, survival time, dopamine neurons, and peripheral neurons, as well as the NAD+ levels in the MPTP-affected PD zebrafish model. In addition, transcriptome sequencing analysis revealed that, after NR treatment, differentially expressed genes were significantly enriched in the glucose metabolism and protein processing pathways in the endoplasmic reticulum (ER). Quantitative PCR (qPCR) revealed that the mRNA levels of the glycoheterotrophic enzyme (involved in glucose metabolism) were significantly decreased, and the glycolytic enzyme mRNA expression levels were significantly increased. The results of the non-targeted metabolomic analysis showed that NR treatment significantly increased the levels of metabolites such as nicotinic acid ,nicotinamide, d-glucose (from the gluconeogenesis and glycolysis metabolism pathways) and some glucogenic amino acids, such as glutamine. Importantly, NR ameliorated MPTP-induced endoplasmic reticulum stress (ERS) in the PD zebrafish model through the Perk-Eif2α-Atf4-Chop pathway. These results highlight the neuroprotective effect of NR in the present PD zebrafish model through modulation of glucose metabolism and ERS via the Perk-Eif2α-Atf4-Chop pathway and provide valuable mechanistic insights into the treatment of PD.

Dihydroartemisinin modulated arachidonic acid metabolism and mitigated liver inflammation by inhibiting the activation of 5-LOX and COX-2

BackgroundDihydroartemisinin (DHA), a derivative of Artemisia annua, has been shown to possess anti-inflammatory properties. Besides, Yes-associated protein 1 (YAP1) plays a crucial role in maintaining liver homeostasis. MethodsThis study used Yap1Flox/Flox, Albumin-Cre mice with hepatocyte-specific Yap1 knockout (referred to as Yap1LKO) and their control mice (Yap1Flox/Flox, referred to as Yap1Flox). The effect of Yap1 on lipid metabolism homeostasis was investigated through non-targeted metabolomic analysis of mouse liver. Subsequently, DHA was administered to Yap1LKO mice to assess its potential as a treatment. Liver pathology was evaluated via H&E staining, and the levels of AST, ALT, and TG were quantified using biochemical assays. The contents of arachidonic acid (AA), prostaglandin E1 (PGE1), and leukotrienes (LT) in the liver were measured using ELISA, while the protein expressions of PLIN2, 5-lipoxygenase (5-LOX), and cyclooxygenase-2 (COX-2) were analyzed through IHC staining. ResultsHepatocyte-specific Yap1 knockout activated the AA metabolic pathway, resulting in increased elevated levels of AA, PGE1, and LT levels, along with inflammatory cytokine infiltration. DHA mitigated the elevation of metabolites such as PGE1 and LT caused by the AA metabolic pathway activation by down-regulating the levels of COX-2 and 5-LOX in the liver of Yap1LKO mice. Moreover, it alleviated the accumulation of lipid vacuoles and reduced triglyceride (TG) and perilipin-2 (PLIN2) levels in the liver of Yap1LKO mice. ConclusionsExcessively low YAP1 expression induces liver inflammation and disturbances in lipid metabolism, whereas DHA modulated AA metabolism and mitigated liver inflammation by inhibiting the activation of 5-LOX and COX-2.

Quality improvement of soybean meal by simultaneous microbial fermentation and enzymolysis and untargeted metabolomic analysis of its metabolites

Twelve lactobacillus strains, four yeast strains, and eight proteases were screened, and their effects on oligosaccharide degradation, lactic acid production, and proteolytic capacity were investigated in soybean meal. The simultaneous microbial fermentation and enzymolysis (SMFE) conditions were optimized as feed/water ratio of 1:0.8 (w/v), yeast inoculum of 2.1×105 CFU/g, lactobacillus inoculum of 2.1×107 CFU/g, protease addition of 200 U/g, and fermentation at 37 °C for 48 h. The results showed that SMFE soybean meal exhibited superior performance over unprocessed soybean meal, showing increases in crude protein content from 49.8% to 56.3%, acid-soluble protein content from 3.7% to 10.4%, lactic acid production from 0 to 71.5 g/kg, and a decrease in oligosaccharide content from 6.4% to 0%. The comprehensive performance of SMFE soybean meal is higher than that from enzymolysis or fermentation alone Non-targeted metabolomic analysis revealed a diversity of metabolites in fermented soybean meal produced by different processes, and 8366 detected metabolites were mainly amino acids and their metabolites. Furthermore, among the six experimental groups, the metabolites of the SMFE group, and the yeast and lactic acid bacteria fermentation groups were the most similar. This study facilitates the assessment of overall quality of soybean meal and reveals the superiority of SMFE over traditional methods in enhancing nutritional profile of soybean meal.

TMT-based quantitative proteomics and non-targeted metabolomic analyses reveal the inactivation mechanism of cold atmospheric plasma against Pseudomonas aeruginosa

Cold atmospheric plasma (CAP) has emerged as a potent nonthermal inactivation strategy in food processing over the past two decades. However, the underlying mechanisms of inactivation remain unclear. In this study, Tandem Mass Tag (TMT)-based quantitative proteomics and non-targeted metabolomic analyses were conducted to investigate the responses of Pseudomonas aeruginosa to CAP. The results revealed significant alterations in 170 differentially expressed proteins (DEPs) and 490 differential metabolites (DMs) upon air-CAP treatment. P. aeruginosa demonstrated a regulatory response at the transcription and translation levels, upregulating proteins to resist external stimuli. Conversely, a predominant down-regulation of proteins indicated that CAP treatment profoundly disrupted the cell structure, inhibiting movement, colonization ability, virulence protein secretion, and bacterial biofilm formation. Moreover, CAP compromised the bacterium's ability to acquire energy, thereby disrupting its defense mechanisms, reducing drug resistance, and potentially leading to bacterial death. This comprehensive study enhances our understanding of the mode of action of air-CAP against P. aeruginosa. The findings provide a robust experimental foundation for considering CAP as an effective inactivation method in the food industry.

Effect of SlSAHH2 on metabolites in over-expressed and wild-type tomato fruit.

Tomato (Solanum lycopersicum) is an annual or perennial herb that occupies an important position in daily agricultural production. It is an essential food crop for humans and its ripening process is regulated by a number of genes. S-adenosyl-l-homocysteine hydrolase (AdoHcyase, EC 3.3.1.1) is widespread in organisms and plays an important role in regulating biological methylation reactions. Previous studies have revealed that transgenic tomato that over-express SlSAHH2 ripen earlier than the wild-type (WT). However, the differences in metabolites and the mechanisms driving how these differences affect the ripening cycle are unclear. To investigate the effects of SlSAHH2 on metabolites in over-expressed tomato and WT tomato. SlSAHH2 over-expressed tomato fruit (OE-5# and OE-6#) and WT tomato fruit at the breaker stage (Br) were selected for non-targeted metabolome analysis. A total of 733 metabolites were identified by mass spectrometry using the Kyoto Encyclopedia of Genes and Genomes (KEGG) database and the Human Metabolome database (HMDB). The metabolites were divided into 12 categories based on the superclass results and a comparison with the HMDB. The differences between the two databases were analyzed by PLS-DA. Based on a variable important in projection value >1 and P < 0.05, 103 differential metabolites were found between tomato variety OE-5# and WT and 63 differential metabolites were found between OE-6# and WT. These included dehydrotomatine, L-serine, and gallic acid amongst others. Many metabolites are associated with fruit ripening and eight common metabolites were found between the OE-5# vs. WT and OE-6# vs. WT comparison groups. The low L-tryptophan expression in OE-5# and OE-6# is consistent with previous reports that its content decreases with fruit ripening. A KEGG pathway enrichment analysis of the significantly different metabolites revealed that in the OE-5# and WT groups, up-regulated metabolites were enriched in 23 metabolic pathways and down-regulated metabolites were enriched in 11 metabolic pathways. In the OE-6# and WT groups, up-regulated metabolites were enriched in 29 pathways and down-regulated metabolites were enriched in six metabolic pathways. In addition, the differential metabolite changes in the L-serine to flavonoid transformation metabolic pathway also provide evidence that there is a phenotypic explanation for the changes in transgenic tomato. The metabolomic mechanism controlling SlSAHH2 promotion of tomato fruit ripening has been further elucidated.

USP9X-mediated REV1 deubiquitination promotes lung cancer radioresistance via the action of REV1 as a Rad18 molecular scaffold for cystathionine γ-lyase.

Radioresistance is a key clinical constraint on the efficacy of radiotherapy in lung cancer patients. REV1 DNA directed polymerase (REV1) plays an important role in repairing DNA damage and maintaining genomic stability. However, its role in the resistance to radiotherapy in lung cancer is not clear. This study aims to clarify the role of REV1 in lung cancer radioresistance, identify the intrinsic mechanisms involved, and provide a theoretical basis for the clinical translation of this new target for lung cancer treatment. The effect of targeting REV1 on the radiosensitivity was verified by in vivo and in vitro experiments. RNA sequencing (RNA-seq) combined with nontargeted metabolomics analysis was used to explore the downstream targets of REV1. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to quantify the content of specific amino acids. The coimmunoprecipitation (co-IP) and GST pull-down assays were used to validate the interaction between proteins. A ubiquitination library screening system was constructed to investigate the regulatory proteins upstream of REV1. Targeting REV1 could enhance the radiosensitivity in vivo, while this effect was not obvious in vitro. RNA sequencing combined with nontargeted metabolomics revealed that the difference result was related to metabolism, and that the expression of glycine, serine, and threonine (Gly/Ser/Thr) metabolism signaling pathways was downregulated following REV1 knockdown. LC-MS/MS demonstrated that REV1 knockdown results in reduced levels of these three amino acids and that cystathionine γ-lyase (CTH) was the key to its function. REV1 enhances the interaction of CTH with the E3 ubiquitin ligase Rad18 and promotes ubiquitination degradation of CTH by Rad18. Screening of the ubiquitination compound library revealed that the ubiquitin-specific peptidase 9 X-linked (USP9X) is the upstream regulatory protein of REV1 by the ubiquitin-proteasome system, which remodels the intracellular Gly/Ser/Thr metabolism. USP9X mediates the deubiquitination of REV1, and aberrantly expressed REV1 acts as a scaffolding protein to assist Rad18 in interacting with CTH, promoting the ubiquitination and degradation of CTH and inducing remodeling of the Gly/Ser/Thr metabolism, which leads to radioresistance. A novel inhibitor of REV1, JH-RE-06, was shown to enhance lung cancer cell radiosensitivity, with good prospects for clinical translation.

Research on the effects of intestinal FXR agonists and antibiotics on the regulation of red kidney bean polysaccharides in the liver metabolism in mice with type 2 diabetes

AbstractOur previous study revealed that red kidney bean polysaccharides (RKB) exhibit a pronounced hypoglycemic effect on type 2 diabetic rats, while simultaneously exerting a significant ameliorative impact on hepatic damage in these animals. However, the precise mechanism underlying the effects of RKB on diabetes and liver metabolism remains unproven. In this study, we utilized a mouse model of type 2 diabetes induced by a high‐fat diet combined with streptozotocin to investigate the impact of RKB. We administered a combined intervention involving antibiotics, fexaramine, and RKB to elucidate the mechanism underlying RKB's effects. Our findings demonstrated that RKB significantly ameliorated liver function indices and histopathological injuries. Nevertheless, when antibiotics and fexaramine were introduced as interventions, they hindered the beneficial effects of RKB on liver function in type 2 diabetic mice. Furthermore, our nontargeted metabolomics analysis revealed that antibiotics and fexaramine exerted their inhibitory actions on RKB efficacy through modulation of distinct metabolites involved in glycerophospholipid and purine metabolic pathways.

Gouqi-derived nanovesicles (GqDNVs) inhibited dexamethasone-induced muscle atrophy associating with AMPK/SIRT1/PGC1α signaling pathway

With the increasing trend of global aging, sarcopenia has become a significant public health issue. Goji berry, also known as “Gou qi zi” in China, is a traditional Chinese herb that can enhance the structure and function of muscles and bones. Otherwise, previous excellent publications illustrated that plant-derived exosome-like nanoparticles can exert good bioactive functions in different aging or disease models. Thus, we issued the hypothesis that Gouqi-derived nanovesicles (GqDNVs) may also have the ability to improve skeletal muscle health, though the effect and its mechanism need to be explored. Hence, we have extracted GqDNVs from fresh berries of Lycium barbarum L. (goji) and found that the contents of GqDNVs are rich in saccharides and lipids. Based on the pathway annotations and predictions in non-targeted metabolome analysis, GqDNVs are tightly associated with the pathways in metabolism. In muscle atrophy model mice, intramuscular injection of GqDNVs improves the cross-sectional area of the quadriceps muscle, grip strength and the AMPK/SIRT1/PGC1α pathway expression. After separately inhibiting AMPK or PGC1α in C2C12 cells with dexamethasone administration, we have found that the activated AMPK plays the chief role in improving cell proliferation induced by GqDNVs. Furthermore, the energy-targeted metabolome analysis in the quadriceps muscle demonstrates that the GqDNVs up-regulate the metabolism of amino sugar and nucleotide sugar, autophagy and oxidative phosphorylation process, which indicates the activation of muscle regeneration. Besides, the Spearman rank analysis shows close associations between the quality and function of skeletal muscle, metabolites and expression levels of AMPK and SIRT1. In this study, we provide a new founding that GqDNVs can improve the quality and function of skeletal muscle accompanying the activated AMPK/SIRT1/PGC1α signaling pathway. Therefore, GqDNVs have the effect of anti-aging skeletal muscle as a potential adjuvant or complementary method or idea in future therapy and research.Graphical

Bioconcentration, oxidative stress and molecular mechanism of the toxic effect of acetamiprid exposure on Xenopus laevis tadpoles

Acetamiprid is a neonicotinoid commonly detected in aquatic ecosystems, with residual concentrations of up to 0.41 mg/L in surface water, posing a threat to the health of nontarget aquatic organisms. However, studies on the potential toxicity and underlying mechanisms of action of acetamiprid on nontarget aquatic organisms are limited. This study investigated the acute and short-term toxicity of acetamiprid to Xenopus laevis tadpoles. A 96-h acute toxicity test determined the LC50 of acetamiprid to be 32.1 mg/L. After 28 days of exposure to 1/10 and 1/100 LC50 concentrations, tadpole samples were collected for bioconcentration elimination analysis, biochemical analyses, transcriptomics, and metabolomics studies to comprehensively evaluate the toxic effects of acetamiprid and its underlying mechanisms. The results, indicating bioconcentration factors (BCFs) < 1, suggest that acetamiprid has a low bioconcentration in tadpoles. Additionally, oxidative stress was observed in treated Xenopus laevis tadpoles. Transcriptomic and nontargeted metabolomic analyses identified 979 differentially expressed genes (DEGs) and 95 differentially metabolites in the 0.321 mg/L group. The integrated analysis revealed that disruption of purine and amino acid metabolic pathways potentially accounts for acetamiprid-induced toxic effects in tadpoles. The disruptive effects of acetamiprid on valine, leucine and isoleucine biosynthesis; and aminoacyl-tRNA biosynthesis metabolic pathways in tadpoles were validated through targeted metabolomics analysis. These findings are crucial for assessing the risk of acetamiprid to nontarget aquatic organisms.

Essential Oil of Hymenocrater longiflorus Benth. as a Pro-Apoptotic Agent in Human Breast Cancer Cells

Background: Breast cancer is one of the most common cancers in women. Current treatments, such as chemotherapy and radiation therapy, often involve numerous side effects. Consequently, it is essential to discover new anticancer compounds that are safer and more effective. Objectives: The aim of this research is to investigate the anticancer effects of Hymenocrater longiflorus Benth. essential oil. Methods: The anticancer properties of H. longiflorus Benth. essential oil were investigated on MDA-MB-231 and MCF-7 breast cancer cells using MTT assay, double staining with acridine orange/ethidium bromide, and flow cytometry. The essential oil of H. longiflorus Benth. was analyzed using gas chromatography-mass spectrometry (GC-MS). Results: The essential oil significantly reduced the viability of breast cancer cells depending on the concentration (dose) and duration. Results from double fluorescent staining and flow cytometric analysis also indicated that apoptosis is the primary mechanism of cell death. Non-targeted GC-MS metabolomic data analysis of the essential oil revealed the presence of various anticancer compounds, including α-pinene, linalool, and caryophyllene, in the essential oil. Conclusions: In conclusion, the essential oil of H. longiflorus Benth. is a promising source of anticancer metabolites that can be further evaluated for drug therapy or as a co-treatment with conventional chemotherapeutic agents.

Protective effect of soluble dietary fiber from Rosa roxburghii Tratt residue on dextran sulfate sodium-induced ulcerative colitis by regulating serum metabolism and NF-κB pathway in mice.

Ulcerative colitis (UC) refers to an idiopathic chronic inflammatory bowel disease that starts with inflammation of the intestinal mucosa. Dietary fiber plays a crucial role in maintaining the normal architecture of the intestinal mucosa. In this study, the protective effect and potential mechanism of soluble dietary fiber from Rosa roxburghii Tratt residue (SDFR) on dextran sulfate sodium (DSS)-induced UC mice were explored. The results revealed that SDFR could ameliorate body weight loss and pathological injury, improve the structure and crypt destruction in colon in DSS-induced mice. Moreover, the levels of NO, IL-1β, TNF-α, MPO and protein expression of iNOS and COX-2 were decreased after administration of SDFR. Notably, nontargeted metabolomics analysis indicated that there were significant differences in 51 potential metabolites in serum between the DSS and control groups. SDFR intervention could regulate aberrant alterations of these metabolites and mitigate UC via regulating metabolic pathways, including arachidonic acid and glycerophospholipid metabolism. This study provides novel evidence that SDFR could be used as a potential modulator to relieve UC. Also, the results provide a theoretical basis for the utilization of byproducts in Rosa roxburghii Tratt fruit processing. © 2024 Society of Chemical Industry.

Down-regulation of O-GlcNAcylation alleviates insulin signaling pathway impairment following arsenic exposure via suppressing the AMPK/mTOR-autophagy pathway

Impairment of the insulin signaling pathway is a key contributor to insulin resistance under arsenic exposure. Specifically, O-GlcNAcylation, an important post-translational modification, plays a crucial role in insulin resistance. Nevertheless, the concrete effect and mechanism of O-GlcNAcylation in arsenic-induced impairment of the insulin signaling pathway remain elusive. Herein, C57BL/6 mice were continuously fed arsenic-containing food, with a total arsenic concentration of 30 mg/kg. We observed that the IRS/Akt/GSK-3β insulin signaling pathway was impaired, and autophagy was activated in mouse livers and HepG2 cells exposed to arsenic. Additionally, O-GlcNAcylation expression in mouse livers and HepG2 cells was elevated, and the key O-GlcNAcylation homeostasis enzyme, O-GlcNAc transferase (OGT), was upregulated. In vitro, non-targeted metabolomic analysis showed that metabolic disorder was induced, and inhibition of O-GlcNAcylation restored the metabolic profile of HepG2 cells exposed to arsenic. In addition, we found that the compromised insulin signaling pathway was dependent on AMPK activation. Inhibition of AMPK mitigated autophagy activation and impairment of insulin signaling pathway under arsenic exposure. Furthermore, down-regulation of O-GlcNAcylation inhibited AMPK activation, thereby suppressing autophagy activation, and improving the impaired insulin signaling pathway. Collectively, our findings indicate that arsenic can impair the insulin signaling pathway by regulating O-GlcNAcylation homeostasis. Importantly, O-GlcNAcylation inhibition alleviated the impaired insulin signaling pathway by suppressing the AMPK/mTOR-autophagy pathway. This indicates that regulating O-GlcNAcylation may be a potential intervention for the impaired insulin signaling pathway induced by arsenic.

PSVI-18 Low and high feed efficiency grower-finisher pigs fed three different levels of dietary crude protein showed no immediate difference in blood metabolomics profile

Abstract Improving feed efficiency (FE) of grower-finisher pigs is of great importance considering its impact on production economy, nutrient excretions as well as carbon-footprint. The biological background describing variation in FE between pen mates, is difficult to establish as feed intake (FI) and FE are not estimated on an individual level. In this study advanced feeding stations were used to estimate FI and further calculate feed conversion ratio (FCR) on individual level. The study aimed to investigate the possible difference in the blood metabolic profile between low and high FE grower-finisher pigs fed three different levels of dietary crude protein (CP). The study included 60 newly weaned gilts divided in two series, housed as 10 pigs/pen. The 30 pigs per series were distributed between three pens (10 pigs/pen) and were balanced by body weight and litter. Throughout the entire weaner period all pigs had ad libitum access to the same diet. Pigs were redistributed among the grower-finisher pens to balance FCR levels between dietary treatments (CP levels) and to reduce any carry over effect from the weaner to the grower-finisher pens. From 30-115 kg body weight (BW), pigs were fed either a low (L; 14.8% CP), standard (15.2% CP) or high (H; 15.8% CP) CP diet equal in energy, where BW, average weekly gain (AWG), and FCR were determined on weekly basis, while FI was estimated on daily basis. Three low FE (20% least) and three high FE (20% greatest) pigs were selected based on the final 4 wk of growth within station and CP level. On the final day of the experimental period all low-FE and high-FE pigs within each dietary treatment were euthanized and blood plasma was sampled and subjected to a nontargeted LC-MS metabolomics analysis. Growth performance and metabolite data were, respectively, analyzed in R and MetaboAnalyst 5.0 with FE and CP as fixed effects. High-FE and H-CP pigs continuously displayed numerically greater BW, while AWG and FCR were, respectively, numerically greater, and less in L-CP than H-CP group during last 2 wk of the experimental period. Principal Component Analysis (PCA) showed no grouping (P &amp;gt; 0.05) of metabolites and no specific pattern of the metabolic profile from either the two FE categories or the three dietary CP levels. Furthermore, no correlation (P &amp;gt;0.05) of metabolites to any other observation was found, and only little variation, measured by distance to model showing deviation of each pig to overall model, was observed. In conclusion, at first glance the metabolomics profile did not appear to present any specific pattern between low and high feed efficiency grower-finisher pigs or between pigs exposed to different levels of dietary CP in the grower finisher period.

297 Effects of plasma zinc concentration on longissimus thoracis metabolic status of beef steers

Abstract The objective of this study was to examine the role of Zn status on muscle glucose and other metabolites. Angus steers [n = 144; body weight (BW) = 525 ± 30 kg] with varying plasma Zn concentration and implant status served as the initial pool for this secondary experiment. Steers were assigned to one of two implant (IMP) treatments: no implant (NO) or Component TE-200 (TE-200; Elanco, Greenfield, IN) on d 0. Zinc was supplemented as ZnSO4 at 0 mg Zn/kg dry matter (DM; analyzed 54 mg Zn/kg DM), 30 mg Zn/kg DM, or 100 mg Zn/kg DM, starting 60 d before implant day. Steers were fed via GrowSafe bunks (GrowSafe Systems Ltd., Airdie, AB, Canada), and steer was experimental unit. Steers were fed in two blocks with a 14-d stagger for sample logistics. Blood and muscle biopsies of the longissimus thoracis were collected on d 40 after implant, a minimum of 2 h post-feed delivery. Plasma Zn was quantified via ICP-OES and 144 samples were stratified into quantiles by plasma Zn concentration. Samples (n = 12 low and 12 high from NO and TE-200 groups) were identified and designated as low plasma Zn (LOW, 1.1 mg Zn/L average) and high plasma Zn (HI, 1.6 mg Zn/L average) treatments (TRT). Muscle samples from these 48 steers were homogenized and derivatized for subsequent analysis via gas chromatography-mass spectrometry (GCMS) for non-targeted metabolomic analysis. Data were analyzed as a complete randomized design using the MIXED procedure of SAS 9.4 (SAS Inst. Inc., Cary, NC) with fixed effects of TRT, IMP, BLOCK, and TRT×IMP. A TRT×IMP effect was observed for muscle glycine (P = 0.02) where LOW/IMP was similar to HI/NO but had greater glycine concentrations than LOW/NO and HI/IMP. There was a tendency for a TRT×IMP (P = 0.07) effect where LOW/IMP and HI/NO tended to have greater concentrations of hydroxyisovaleric acid. Given the role of glycine in protein synthesis and hydroxyisovaleric acid in leucine metabolism these data suggest differential protein responses to implants in cattle of lesser vs. greater plasma Zn concentrations. Hydroxybutyric acid, a ketone body which can be utilized for energy, tended to be greater in muscle of steers receiving an IMP (P = 0.09). Contrary to our hypothesis that greater plasma Zn would support differential muscle glucose metabolism, no effects of plasma Zn TRT or IMP were noted for glucose, lactate, and other detected metabolites. Both LOW and HI plasma Zn concentrations were still considered adequate, and future research should explore the impact of distinctly deficient and adequate plasma Zn concentration on glucose metabolism pathways in muscle.

The impact of continuous cultivation of Ganoderma lucidum on soil nutrients, enzyme activity, and fruiting body metabolites

To explore the impacts of continuous Ganoderma lucidum cultivation on soil physicochemical factors, soil enzyme activity, and the metabolome of Ganoderma lucidum fruiting bodies, this study conducted two consecutive years of cultivation on the same plot of land. Soil physicochemical factors and enzyme activity were assessed, alongside non-targeted metabolomic analysis of the Ganoderma lucidum fruiting bodies under continuous cultivation. The findings unveiled that in the surface soil layer (0–15 cm), there was a declining trend in organic matter, ammonium nitrogen, available phosphorus, available potassium, pH, polyphenol oxidase, peroxidase, alkaline phosphatase, and sucrase, whereas nitrate nitrogen, electrical conductivity (EC), and salt content exhibited an upward trend. Conversely, in the deeper soil layer (15–30 cm), organic matter, ammonium nitrogen, available potassium, alkaline phosphatase, and sucrase demonstrated a decreasing trend, while nitrate nitrogen, available phosphorus, pH, EC, salt content, polyphenol oxidase, and soil peroxidase showed an increasing trend. Metabolomic analysis of Ganoderma lucidum fruiting bodies distinguished 64 significantly different metabolites between the GCK and GT groups, with 39 components having markedly higher relative contents in GCK and 25 components having significantly lower relative contents in GCK compared to GT. Moreover, among these metabolites, there were more types with higher contents in the fruiting bodies harvested in the first year (GCK) compared to those harvested in the second year (GT), with pronounced differences. KEGG pathway analysis revealed that GCK exhibited more complex metabolic pathways compared to GT. The metabolites of Ganoderma lucidum fruiting bodies were predominantly influenced by soil physicochemical factors and soil enzyme activity. In the surface soil layer (0–15 cm), the metabolome was significantly affected by soil pH, soil organic matter, available phosphorus, and soil alkaline phosphatase, while in the deeper soil layer (15–30 cm), differences in the Ganoderma lucidum metabolome were more influenced by soil alkaline phosphatase, soil catalase, pH, nitrate nitrogen, and soil sucrase.

Gas chromatography-mass spectrometry and non-targeted metabolomics analysis reveals the flavor and nutritional metabolic differences of cow's milk fermented by Lactiplantibacillus plantarum with different phenotypic

This study comprehensively evaluated the effects of Lactiplantibacillus plantarum T1 and CSK (ropy and non-ropy phenotypes), on flavour, and nutrient metabolites of fermented milks using gas chromatography-mass spectrometry and non-targeted metabolomics. The microrheological analysis showed that the protein gelation time of CSK fermented milk was earlier (5.2 h) than that of T1 (7.0 h), while the fermented milk by T1 had higher elastic modulus. Fermentation by both strains significantly enhanced the antioxidant activity of the fermented milk, including scavenging capabilities against ABTS, DPPH, and hydroxyl radicals. Meanwhile, fermentation by two strains produced various volatile flavor compounds (hexanoic acid, caprylic acid, capric acid, nonanal, decanal, 2-heptanone and 2-nonanone, etc.), and L. plantarum CSK displayed more stronger acid production capacity. In addition, a total of 397 shared metabolites and 81 unique metabolites were identified by untargeted metabolomics analysis of the two groups of fermented milks, mainly including lipids and lipid-like molecules (66, 27.27%), organic heterocyclic compounds (40, 16.53%), phenylketones and polyketones (23, 9.5%), and organic acids and their derivatives (49, 20.25%). Furthermore, both strains significantly elevated the amino acid nutritional levels in the fermented milk, conferred an abundance of active metabolites such as niacin and nicotinamide. During the fermentation, L. plantarum CSK notably produced antioxidant fumaric acid and exhibited higher fatty acid metabolism compared to L. plantarum T1. Overall, this study provides some practical basis for the development of fermented cow's milk with good antioxidant activity and high nutritional quality using different phenotypes of L. plantarum.