PubMed

Environ Pollut. 2022 Nov 9:120635. doi: 10.1016/j.envpol.2022.120635. Online ahead of print.ABSTRACTDue to the wide application of plastic products in human life, microplastic pollution in water has recently attracted more attention. Many studies have revealed the size-dependent toxicity of microplastics. Here, we investigated the toxicological effects of polystyrene microplastics (PS-MPs) on the white leg shrimp, Litopenaeus vannamei, a profitable aquaculture species, using a comprehensive histomorphological, microbiome, and metabolomic approach to verify whether smaller particles are more toxic than larger particles. L. vannamei were experimentally exposed to water containing PS-MPs of four sizes (0.1, 1.0, 5.0, and 20.0 μm) for 24 h at 10 mg/L (acute experiment) and 12 d at 1 mg/L (subchronic experiment). After 24 h of acute exposure, PS-MP accumulation in shrimp indicated that the ingestion and egestion of PS-MPs had a size-dependent effect, and smaller particles were more bioavailable. The tissue morphological results of subchronic experiments showed that, for the guts and gills, the smaller sizes of the PS-MPs exhibited greater damage. In addition, 16 S rRNA gene amplicon sequencing showed that the alpha diversity was higher under larger PS-MP exposure. Correlated with changes in intestinal bacteria, we found a greater enrichment of metabolic pathways in hemolymph proteins and metabolites in larger PS-MP groups, such as "arginine and proline metabolism", "protein digestion and absorption", "lysine degradation". Interestingly, the activity or content of biomarkers of oxidative stress showed a peak at 1 μm and 5 μm. Under specific sizes of PS-MPs, the abundance of the pathogen Vibrio and probiotic bacteria Rhodobacter (5-μm) and Bacillus and Halomonas (1-μm) were simultaneously enriched. Our results indicated that PS-MP exposure can cause size-dependent damage to shrimp, yet specific particle size can be influential differently in regard to some research indicators. Therefore, it can enhance our comprehensive understanding of the impacts of microplastics on shrimp health and suggests that specific particle size should be considered when assessing the size-dependent toxicity of microplastics.PMID:36370970 | DOI:10.1016/j.envpol.2022.120635

Environ Pollut. 2022 Nov 9:120626. doi: 10.1016/j.envpol.2022.120626. Online ahead of print.ABSTRACTModern agriculture depends highly on pesticides and pharmaceutical preparations, so controlling exposure to these substances in the feed and food chain is essential. This article presents the first study on residues of a broad spectrum of pesticides and veterinary drugs in the diets of dairy cattle. One hundred and two representative samples of the complete diets, including basal feed rations and additional fed concentrate, were collected in three Austrian provinces (Styria, Lower and Upper Austria) in 2019 and 2020. The samples were tested for >700 pesticides, veterinary drugs and related metabolites using a validated method based on liquid chromatography/electrospray ionization-tandem mass spectrometry (LC/ESI-MS/MS). In total, 16 residues (13 pesticides and three veterinary drug residues) were detected. > 90% of the diets contained pesticide residues and <10% veterinary drug residues, whereas banned pesticides were not found. The most frequent pesticide residues were fluopyram (62%), piperonyl butoxide (39%) and diethyltoluamide (35%). The following pesticides exceed the default EU maximum residue level (MRL) (10 μg kg-1) for products exclusively used for animal feed production: Benzovindiflupyr (proportion of samples > MRLs: 1%), bixafen (2%), fluopyram (6%), ipconazole (1%) and tebuconazole (3%). Three residues (dinitrocarbanilide, monensin and nicarbazin) of veterinary drugs were identified, all below the MRLs. Over 60% of the evaluated samples contained mixtures of two to six residues/sample. Only one pesticide (diethyltoluamide) presented a significant difference among regions, with higher concentrations in Upper Austria. Brewery's spent grains were the dietary ingredient that showed the strongest correlation to pesticide residues. These findings evidence the realistic scenario of highly occurrent low doses of pesticides cocktails in the feed/food chain, which may affect the animal, human and environmental health. Since the risk assessments are based on single pesticides, the potential synergistic effect of co-occurring chemicals ("cocktail effect") requires further investigations.PMID:36370968 | DOI:10.1016/j.envpol.2022.120626

Free Radic Biol Med. 2022 Nov 9:S0891-5849(22)00969-8. doi: 10.1016/j.freeradbiomed.2022.11.007. Online ahead of print.ABSTRACTBACKGROUND: The joint influence of metabolic patterns on bone fragility has been rarely evaluated. We assessed the prospective association of plasma metabolic patterns with incident osteoporosis-related bone fractures in the Hortega Study participants. Redox balance plays a key role in bone metabolism. We also assessed differential associations in participant subgroups by redox-related metal exposure levels and candidate genetic variants.MATERIAL AND METHODS: In 467 participants older than 50 years from the Hortega Study, a representative sample from a region in Spain, we estimated metabolic principal components (mPC) for 54 plasma metabolites from NMR-spectrometry. BMD was calculated in the right calcaneus using Peripheral Instantaneous X-ray Imaging system. Metals biomarkers were measured in plasma by AAS and in urine by HPLC-ICPMS. Redox-related SNPs (N = 341) were measured by oligo-ligation assay.RESULTS: The prospective association with incident bone fractures was inverse for mPC1 (non-essential and essential amino acids, including branched-chain, and bacterial co-metabolites, including isobutyrate, trimethylamines and phenylpropionate, versus fatty acids and VLDL) and mPC4 (HDL), but positive for mPC2 (essential amino acids, including aromatic, and bacterial co-metabolites, including isopropanol and methanol). Participants with decreased selenium and increased antimony, arsenic and, suggestively, cadmium exposures showed higher mPC2-associated bone fractures risk. Genetic variants annotated to 19 genes, with the strongest evidence for NCF4, NOX4 and XDH, showed differential metabolic-related bone fractures risk.CONCLUSIONS: Bone was influenced by metabolic patterns associated to amino acids, microbiota co-metabolism and lipid metabolism. Carriers of redox-related variants may benefit from metabolic interventions to prevent the bone frailty consequences based on their antimony, arsenic, selenium, and, possibly, cadmium, exposure levels.PMID:36370960 | DOI:10.1016/j.freeradbiomed.2022.11.007

J Hepatol. 2022 Nov 9:S0168-8278(22)03288-3. doi: 10.1016/j.jhep.2022.10.031. Online ahead of print.ABSTRACTBACKGROUND AND AIMS: Acute liver failure (ALF) is a life-threatening disease characterised by high-grade inflammation and immunoparesis with a high incidence of death from sepsis. Here, we aimed to describe the metabolic dysregulation in ALF and determine whether systemic immune responses are modulated via the lysophosphatidylcholine(LPC)-autotaxin(ATX)-lysophosphatidylcholinic acid (LPA) pathway.METHODS: 96 ALF patients, 71 healthy controls (HC), 104 patients with cirrhosis and 31 septic patients were recruited. The pathways of interest were identified based on multivariate statistical analysis of proton nuclear magnetic resonance (1HNMR) spectroscopy, untargeted ultraperformance liquid chromatography-mass spectrometry (UPLC-MS)-based lipidomics and validated with a targeted metabolomics panel. Peripheral blood mononuclear cells were cultured with LPA 16:0, 18:0, 18:1, and their immune checkpoint surface expression was assessed by flow cytometry. LPA receptor (LPAR) transcript-level expression of monocytes was investigated and the effect of LPAR antagonism was also examined in vitro.RESULTS: LPC 16:0 was found highly discriminant between ALF and HC. There was an increase in ATX and LPA in ALF compared to HC and sepsis. LPCs 16:0, 18:0 and 18:1 were reduced in ALF patients with poor prognosis. Treatment of monocytes with LPA 16:0 increased their PD-L1 expression and reduced CD155, CD163, MerTK levels, without effect on T and NK/CD56+T cells immune checkpoints. LPAR1 and 3 antagonism in culture reversed the LPA effect on monocyte expression of MerTK and CD163. MerTK and CD163, but not LPARs genes, were differently expressed and upregulated in monocytes from ALF patients compared to controls.CONCLUSION: Reduced amounts of LPCs are biomarkers of poor prognosis in patients with ALF. The LPC-ATX-LPA axis appears to modulate innate immune response in ALF via LPAR1 and LPAR3. Further investigations are required to identify novel therapeutic agents targeting these receptors.IMPACT AND IMPLICATIONS: Liver disease is the 5th leading cause of death in the UK and rising in incidence. Acute liver failure occurs on the background of normal liver function and mostly in young adults. Acute admissions to hospital and intensive care units are rising in the UK and worldwide. We identified a metabolic signature of acute liver failure and investigated the immunometabolic role of the Lysophosphatdylcholine(LPC)-Autotaxin (ATX)-Lysophosphatidylcholinic acid (LPA) pathway in order to find a mechanistic explanation for monocyte behaviour and find possible therapeutic target(s) to modulate the systemic immune response in ALF. At present, no selective immune based therapies exist. We were able to modulate monocyte phenotype and function in vitro and aim to extend findings to murine models of ALF before could apply this treatment to patients. Future therapies may be based on the enhancement of resolution through metabolic modulation and therefore the role of specific lipids in this pathway require elucidation and the relative merits of ATX inhibition, LPAR blockade or lipid-based therapies answered. This application aims to make a step change in meeting this knowledge gap and definitively elucidate these immune-metabolic pathways using an experimental medicine approach, thus finding the most effective therapeutic targets.PMID:36370949 | DOI:10.1016/j.jhep.2022.10.031

Bioresour Technol. 2022 Nov 9:128301. doi: 10.1016/j.biortech.2022.128301. Online ahead of print.ABSTRACTMicroalgae cannot meet the bait demand for aquaculture due to light intensity limitation and other disadvantageous conditions. This research selected 6 mixotrophic microalgae, and the optimal strains and organic carbon were screened. The results showed that Thalassiosira pseudonana and Chlorella sp. are suitable for mixotrophic culture. The maximum cell density of Thalassiosira pseudonana was found to be 1.67 times than that of the photoautotrophic group when glycerol was added. The maximum cell density of Chlorella sp. with acetic acid was 1.69 times than that of the photoautotrophic group. When the concentration of acetic acid was 5.0 g·L-1 and the concentration of KNO3 was 0.2 g·L-1, the maximum biomass of Chlorella sp. could reach 3.54×107 cells·mL-1; the maximum biomass of Thalassiosira pseudonana was 5.53×106 cells·mL-1 with 10.0 g·L-1 glycerol and 0.2 g·L-1 KNO3. Metabolomic analysis further revealed that mixotrophic bait microalgae could promote the accumulation of lipids and amino acids.PMID:36370937 | DOI:10.1016/j.biortech.2022.128301

J Nutr Biochem. 2022 Nov 9:109214. doi: 10.1016/j.jnutbio.2022.109214. Online ahead of print.ABSTRACTObservational study indicated that folic acid (FA) supplementation may protect against tuberculosis-drug-induced liver injury (TBLI). The aim is to investigate the effect and mechanism of FA on TBLI in rats. Liver injury was induced by a daily gavage of isoniazid (INH) and rifampicin (RIF) in the model and FA groups. Rats in the FA group were also treated with 2.5 mg/kg body weight FA. Rats in the control group were not treated. Eight rats were used in each group. The severity of liver injury was measured by the serum levels of hepatic enzymes and histological score. The metabolites in serum and liver tissues were analyzed by HPLC-Q-TOF-MS/MS. FA treatment significantly reduced alanine aminotransferase and liver necrosis. Seventy-nine differential metabolites in the serum and liver tissues were identified among the three groups. N-acylethanolamines, INH and RIF metabolites, phosphatidylcholines, lysophosphatidylcholines, monoglycerides, diglycerides and bile acids were regulated by FA treatment, involving key metabolic pathways, such as N-acylethanolamine metabolism, INH and RIF metabolism, liver regeneration, inflammation alleviation and bile acid metabolism. RT-PCR and western blotting results confirmed the altered N-acylethanolamine metabolism and improved drug metabolism by FA. In conclusion, FA was protective against TBLI, which may be related to the regulation of N-acylethanolamine metabolism and drug detoxification by FA.PMID:36370928 | DOI:10.1016/j.jnutbio.2022.109214

Phytochemistry. 2022 Nov 9:113508. doi: 10.1016/j.phytochem.2022.113508. Online ahead of print.ABSTRACTThe hop plant (Humulus lupulus L.) has been exploited for a long time for both its brewing and medicinal uses, due in particular to its specific chemical composition. These last years, hop cultivation that was in decline has been experiencing a renewal for several reasons, such as a craze for strongly hopped aromatic beers. In this context, the present work aims at investigating the genetic and chemical diversity of fifty wild hops collected from different locations in Northern France. These wild hops were compared to ten commercial varieties and three heirloom varieties cultivated in the same sampled geographical area. Genetic analysis relying on genome fingerprinting using 11 microsatellite markers showed a high level of diversity. A total of 56 alleles were determined with an average of 10.9 alleles per locus and assessed a significant population structure (mean pairwise FST = 0.29). Phytochemical characterization of hops was based on volatile compound analysis by HS-SPME GC-MS, quantification of the main prenylated phenolic compounds by UHPLC-UV as well as untargeted metabolomics by UHPLC-HRMS and revealed a high level of chemical diversity among the assessed wild accessions. In particular, analysis of volatile compounds revealed the presence of some minor but original compounds, such as aromadendrene, allo-aromadendrene, isoledene, β-guaiene, α-ylangene and β-pinene in some wild accessions; while analysis of phenolic compounds showed high content of β-acids in these wild accessions, up to 2.37% of colupulone. Genetic diversity of wild hops previously observed was hence supported by their chemical diversity. Sample soil analysis was also performed to get a pedological classification of these different collection sites. Results of the multivariate statistical analysis suggest that wild hops constitute a huge pool of chemical and genetic diversity of this species.PMID:36370882 | DOI:10.1016/j.phytochem.2022.113508

Chemosphere. 2022 Nov 9:137213. doi: 10.1016/j.chemosphere.2022.137213. Online ahead of print.ABSTRACTThe biosynthesis of amino acids (AAs) in plants is affected by different nitrogen (N) sources. The effects of exogenous cyanide (KCN) on the concentrations and profiles of AAs in rice seedlings was carried out in the presence of nitrate (+NO3-)/ammonium (+NH4+) or N deficiency (-N). Targeted metabolomics analysis indicated that the highest accumulation of AAs in CN--treated rice seedlings was detected in the "CN-+NH4+" treatments than in other treatments, wherein the doses of exogenous KCN did not significantly affect the total amount of AAs in rice seedlings at the same N fertilized condition. The total content of AAs in rice shoots under "CN-+NH4+" treatments was higher than other treatments, while the total content of AAs in rice roots under "CN-+NO3-" treatments was higher than other treatments. Also, the profiles of 21 AAs in CN--treated rice seedlings showed tissue-specific under different N fertilization. The relative importance index (RII) of AA was used to evaluate the importance of AAs in CN--treated rice seedlings under different N fertilization. The common AAs with higher RII values were compared between three different treatments of KCN (e.g., 0, 1, and 2 mg CN/L). Under "CN-+(-N)" treatments, Ala, Asp, Glu, Val, and Gly (Ala, Gly, Val, and Lys) were the common AAs in rice roots (shoots). Under "CN-+NO3-" treatments, Ala, Glu, Asp, Ser, and Thr (Asp, Ala, Thr, Ser, and Asn) were the common AAs with higher RII values in rice roots (shoots) between all CN- treatments. Under "CN-+NH4+" treatments, Asp, Gln, Asn, and Ala (Asp, Glu, and Thr) are the common AAs with higher RII values in rice roots (shoots) between all CN- treatments. These results suggested that using the RII to describe the change and fluctuation of AAs in rice plants may reflect the different N utilization strategies in response to exogenous CN- exposure.PMID:36370756 | DOI:10.1016/j.chemosphere.2022.137213

Food Chem. 2022 Nov 2;405(Pt A):134807. doi: 10.1016/j.foodchem.2022.134807. Online ahead of print.ABSTRACTFruit of Rosa roxburghii Tratt (FRR) is widely used in functional food industry while short of metabolites research. Herein, we firstly identified 251 metabolites in FRR based on untargeted liquid chromatography-mass spectrometry (LC-MS) approach. Then, 42 differential compounds were sought out to avoid the confusing use of FRR and fruit of R. sterilis S. D. Shi (FRS), and FRR was evaluated exhibiting higher biofunction potential. Moreover, a quantitative LC-MS approach was established to determine the contents of 3 ascorbyl hexosides, and FRR with higher contents should be better source than FRS. Additionally, 17 ascorbic acid (AA) derivatives formed by conjugation of ascorbyl unit with organic acids, flavonoids, or glucuronic acid were also discovered in FRR through characteristic ions of AA and feature-based molecular networking (FBMN), enlightening that AA derivatives were not limited to ascorbyl glycosides. This study provided abundant metabolites information of FRR, laying the basis for exploitation of FRR.PMID:36370576 | DOI:10.1016/j.foodchem.2022.134807

J Ind Microbiol Biotechnol. 2022 Nov 12:kuac025. doi: 10.1093/jimb/kuac025. Online ahead of print.ABSTRACTNitrogen is a critical nutrient in beverage fermentations, influencing fermentation performance and formation of compounds that affect organoleptic properties of the product. Traditionally, most commercial wine fermentations rely on Saccharomyces cerevisiae but the potential of alternative yeasts is increasingly recognised because of the possibility to deliver innovative products and process improvements. In this regard, Saccharomyces uvarum is an attractive non-traditional yeast that, while quite closely related to S. cerevisiae, displays a different fermentative and aromatic profile. Although S. uvarum is used in cider-making and in some winemaking, better knowledge of its physiology and metabolism is required if its full potential is to be realised. To address this gap, we performed a comparative analysis of the response of S. uvarum and S. cerevisiae to 13 different sources of nitrogen, assessing key parameters such as growth, fermentation performance, the production of central carbon metabolites and aroma volatile compounds. We observed that the two species differ in the production of acetate, succinate, medium-chain fatty acids, phenylethanol, phenylethyl acetate and fusel/branched acids in ways that reflect different distribution of fluxes in the metabolic network. The integrated analysis revealed different patterns of yeast performance and activity linked to whether growth was on amino acids metabolised via the Ehrlich pathway or on amino acids and compounds assimilated through the central nitrogen core. This study highlights differences between the two yeasts and the importance that nitrogen metabolism can play in modulating the sensory profile of wine when using S. uvarum as the fermentative yeast.PMID:36370452 | DOI:10.1093/jimb/kuac025

Methods Mol Biol. 2023;2603:163-171. doi: 10.1007/978-1-0716-2863-8_13.ABSTRACTCultured primary neurons are a well-established model for the study of neuronal function. Conventional stable isotope labeling with amino acids in cell culture (SILAC) requires nearly complete metabolic labeling of proteins and therefore is difficult to apply to cultured primary neurons, which do not divide in culture. In a multiplex SILAC strategy, two different sets of heavy amino acids are used for labeling cells for the different experimental conditions. This allows for straightforward SILAC quantitation using partially labeled cells because the two cell populations are always equally labeled. When combined with bioorthogonal noncanonical amino acid tagging (BONCAT), it allows for comparative proteomic analysis of de novo protein synthesis. Here we describe protocols that utilize the multiplex SILAC labeling strategy for primary cultured neurons to study steady-state and nascent proteomes.PMID:36370278 | DOI:10.1007/978-1-0716-2863-8_13

Appl Microbiol Biotechnol. 2022 Nov 12. doi: 10.1007/s00253-022-12261-x. Online ahead of print.ABSTRACTBacterial pathogenicity is greatly affected by nutrient recognition and utilization in the host microenvironment. The characterization of enteral nutrients that promote intestinal pathogen virulence is helpful for developing new adjuvant therapies and inhibiting host damage. Ethanolamine (EA), as a major component of intestinal epithelial cells and bacterial membranes, is abundant in the intestine. Here, we provide the first demonstration that the critical human and porcine pathogen enterotoxigenic Escherichia coli (ETEC) can utilize EA as a nitrogen source, which affects its virulence phenotype. We found that compared with that in M9 medium (containing NH4Cl), EA inhibited ETEC growth to a certain extent; however, the relative expression levels of virulence-related genes, such as ltA (3.0-fold), fimH (2.9-fold), CfaD (2.6-fold), gspD (3.6-fold), and qesE (1.3-fold), increased significantly with 15 mM EA as a nitrogen source (P < 0.05), and the adhesion efficiency of ETEC to Caco-2 cells increased approximately 4.2-fold. In Caco-2 cells, the relative cell viability decreased from 74.8 to 63.4%, and the transepithelial electrical resistance (TEER) cells decreased to 74.8% with intestinal EA (4 mM). In addition, the relative expression levels of proinflammatory factors, such as TNF-α (3.2-fold), INF-γ (2.9-fold), and IL-1β (1.98-fold), in ETEC-infected Caco-2 cells were significantly upregulated (P < 0.05) under EA exposure; however, the above virulence changes were not found in ΔeutR and ΔeutB ETEC. A gas chromatography-mass spectrometry (GC-MS)-based untargeted metabolomics approach was then employed to reveal EA-induced metabolic reprogramming related to ETEC virulence. The data showed that most metabolites related to carbohydrate, aspartate and glutamate metabolism, shikimic acid metabolism, and serine metabolism in ETEC exhibited a decreasing trend with increases in the EA concentration from 0 to 15 mM, but the branched-chain amino acid (BCAA) levels in ETEC increased in a dose-dependent manner under EA exposure. Our data suggest that the intestinal EA concentration can significantly affect the virulence phenotype, metabolic profile, and pathogenicity of ETEC. KEY POINTS: • ETEC growth and virulence gene expression could be regulated by ethanolamine. • The intestinal concentration of EA promoted the damaging effect of ETEC on the host epithelial barrier. • The promoting effect of EA on ETEC toxicity may be related to BCAA metabolism.PMID:36370159 | DOI:10.1007/s00253-022-12261-x

Clin Pharmacol Ther. 2022 Nov 12. doi: 10.1002/cpt.2794. Online ahead of print.ABSTRACTIntravenous (IV) busulfan doses are often personalized to a target plasma exposure (targeted busulfan) using an individual's busulfan clearance (BuCL). We evaluated whether BuCL could be predicted by a predose plasma panel of 841 endogenous metabolomic compounds (EMCs). In this prospective cohort of 132 hematopoietic cell transplant (HCT) patients, all had samples collected immediately before to busulfan administration (preBU) and 96 had samples collected two weeks before busulfan (2-week-preBU). BuCL was significantly associated with 37 EMCs after univariate linear regression analysis and controlling for false discovery (<0.05) in the 132 preBU samples. In parallel, with preBU samples, we included all 841 EMCs in a Lasso penalized regression which selected 13 EMCs as predominantly associated with BuCL. Then, we constructed a prediction model by estimating coefficients for these 13 EMCs, along with sex, using ordinary least-squares. When the resulting linear prediction model was applied to the 2-week-preBU samples, it explained 40% of the variation in BuCL (adjusted R2 =0.40). Pathway enrichment analysis revealed 18 pathways associated with BuCL. Lysine degradation followed by Steroid Biosynthesis, which aligned with the univariate analysis, were the top two pathways. BuCL can be predicted before busulfan administration with a linear regression model of 13 EMCs. This pharmacometabolomics method should be prioritized over use of a busulfan test dose or pharmacogenomics to guide busulfan dosing. These results highlight the potential of pharmacometabolomics as a precision medicine tool to improve or replace pharmacokinetics to personalize busulfan doses.PMID:36369996 | DOI:10.1002/cpt.2794

Vet Q. 2022 Nov 12:1-18. doi: 10.1080/01652176.2022.2145619. Online ahead of print.ABSTRACTBACKGROUND: The complex etiopathology of retained placenta (RP) and hazards associated with it has made it crucial for researchers and clinical veterinarians to study pathogenesis, early-warning diagnosis, and treatment.OBJECTIVES: This study aimed to screen the potential prognostic markers of RP in dairy cows using plasma metabolomics coupled with clinical laboratory indicators.METHODS: Blood samples were collected from 260 dairy cows at 21, 14, 7, and 0 days before parturition and 7, 14, and 21 days after parturition. Consequently, 10 healthy cows and 10 cows with RP with similar parity, body condition score, and age were included in the study. The changes in clinical laboratory indicators of the enrolled cows from 21 before parturition to 21 days after parturition were assessed. After initial overview of the multivariate statistical data using PCA analysis, the data were subjected to orthogonal partial least-squares discriminant analysis.RESULTS: Compared with cows with RP at 7 days before parturition, the levels of endothelin and 6-keto-prostaglandin F1α were increased in healthy cows while the level of estradiol and progesterone decreased. Adenine dinucleotide phosphate, hypoxanthine, guanine dinucleotide phosphate, inosine monophosphate, and L-arginine were revealed as potential prognostic markers of cows with RP at 7 days before parturition involved in the regulation of taste transduction, purine and glutathione metabolism, and autophagy.CONCLUSIONS: The best period for the early-warning diagnosis of RP in dairy cows is 7 days before parturition, and purine metabolism and autophagy may play a vital role in the occurrence and development of RP in dairy cows.PMID:36369933 | DOI:10.1080/01652176.2022.2145619

Anal Bioanal Chem. 2022 Nov 12. doi: 10.1007/s00216-022-04427-3. Online ahead of print.ABSTRACTMedulloblastoma is a malignancy of the central nervous system that occurs most frequently in childhood and is often difficult to diagnose due to its similarities to conventional imaging findings for other pediatric intracranial tumors such as astrocytomas and ependymomas. The purpose of this study was to identify new metabolites and differential metabolic pathways by analyzing the significantly different metabolites present in the plasma of children with medulloblastoma in comparison with those with other intracranial tumors. Plasma was collected from 37 children with medulloblastoma and 34 children with other intracranial tumors. Targeted and non-targeted metabolomics based on ultra-performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) analyses were performed to determine metabolic changes in pediatric medulloblastomas versus other intracranial tumors. Based on multivariate statistical analysis and regression models, we identified differential metabolites in the plasma and investigated different metabolic pathways. A total of 61 differential metabolites in the plasma of children with medulloblastoma were identified by non-targeted metabolomics analysis. In addition, targeted metabolomics analysis identified four differential amino acids, thus allowing us to establish a diagnostic model for children with medulloblastoma. Metabolic pathway analysis showed that there were significant differences in patients with medulloblastoma in terms of glycerophospholipid and α-linolenic acid metabolism pathways as well as several amino acid metabolism pathways (phenylalanine, tyrosine, and tryptophan biosynthesis). We identified differential profiles of key plasma metabolites between children with medulloblastoma and other forms of intracranial tumor, thus providing a basis for identifying early diagnostic markers of medulloblastoma and new therapeutic targets and strategies.PMID:36369592 | DOI:10.1007/s00216-022-04427-3

NPJ Sci Food. 2022 Nov 11;6(1):52. doi: 10.1038/s41538-022-00168-w.ABSTRACTThis study mainly studied sensory and metabolite migration from the skin to the soup in the boiling process of tilapia skin using content analysis, electronic nose technique, electronic tongue technique, and metabolomics technique based on ultra-high performance liquid chromatography-mass spectrometry/mass spectrometry and gas chromatography-time-of-flight-mass spectrometry. The content changes, flavor changes, taste changes, metabolite numbers and differential metabolite numbers for both tilapia skin and soup mainly occurred in the initial 30 min. Moreover, the initial 10 min was the key period for the metabolite changes in the boiling process. Further, the differential metabolites in these three periods (0-10, 10-30, and 30-60 min) were identified to show the metabolites migration process. Six (adenine, gingerol, terephthalic acid, vanillin, pentanenitrile, and 2-pyrrolidinonede) and seven (butyramide, lysope(0:0/20:4(5z,8z,11z,14z)), lysope(22:6(4z,7z,10z,13z,16z,19z)/0:0), linoleic acid, N-acetylneuraminic acid, L-threose, and benzoin) chemicals were screened out in the differential metabolites of tilapia skin and soup, respectively, with Variable Importance in the Projection of >1 and p value of <0.05. This work would be beneficial to understand the sensory and metabolite migration in the preparation process of fish soup and provided a metabolomic analysis route to analyze metabolites migration in food.PMID:36369181 | DOI:10.1038/s41538-022-00168-w

Nat Commun. 2022 Nov 11;13(1):6845. doi: 10.1038/s41467-022-34228-2.ABSTRACTTargeting the intrinsic metabolism of immune or tumor cells is a therapeutic strategy in autoimmunity, chronic inflammation or cancer. Metabolite repair enzymes may represent an alternative target class for selective metabolic inhibition, but pharmacological tools to test this concept are needed. Here, we demonstrate that phosphoglycolate phosphatase (PGP), a prototypical metabolite repair enzyme in glycolysis, is a pharmacologically actionable target. Using a combination of small molecule screening, protein crystallography, molecular dynamics simulations and NMR metabolomics, we discover and analyze a compound (CP1) that inhibits PGP with high selectivity and submicromolar potency. CP1 locks the phosphatase in a catalytically inactive conformation, dampens glycolytic flux, and phenocopies effects of cellular PGP-deficiency. This study provides key insights into effective and precise PGP targeting, at the same time validating an allosteric approach to control glycolysis that could advance discoveries of innovative therapeutic candidates.PMID:36369173 | DOI:10.1038/s41467-022-34228-2

Anal Chim Acta. 2022 Dec 1;1235:340521. doi: 10.1016/j.aca.2022.340521. Epub 2022 Oct 21.ABSTRACTSample preparation is a labor-intensive and time-consuming procedure, especially for the bioanalysis of small-volume samples with low-abundant analytes. To minimize losses and dilution, sample preparation should ideally be hyphenated to downstream on-line analysis such as liquid chromatography-mass spectrometry (LC-MS). In this study, an automated three-phase electro-extraction (EE) method coupled to machine vision was developed, integrated with a robotic autosampler hyphenated to LC-MS. Eight model compounds, i.e. amitriptyline, clemastine, clomipramine, haloperidol, loperamide, propranolol, oxeladin, and verapamil were utilized for the optimization and evaluation of the automated EE setup. The stability of automated EE was evaluated by monitoring the acceptor droplet size by machine vision and recording the current during EE. A Design of Experiment approach (Box-Behnken design) was utilized to optimize the critical parameters of the EE method, i.e., the ratio of formic acid in the sample to acceptor phase, extraction voltage, and extraction time. The developed quadratic models showed good fitness (p < 0.001, R2 > 0.95). Automated EE could be achieved in less than 2 min with enrichment factors (EF) up to 387 and extraction recoveries (ER) up to 97% for academic samples. Finally, the optimized EE method was successfully applied to both spiked human urine and plasma samples with low-concentration (50 ng mL-1) analytes and a low starting sample volume of 20 μL of plasma and urine in 10-fold diluted samples. The developed automated EE setup is easy to operate, provides a fast extraction method for analytes from volume-limited biological samples, and is hyphenated with on-line LC-MS analysis. Therefore, this method can provide fast and automated sample preparation to solve bottlenecks in high-throughput bioanalysis workflows.PMID:36368820 | DOI:10.1016/j.aca.2022.340521

J Hepatol. 2022 Nov 9:S0168-8278(22)03285-8. doi: 10.1016/j.jhep.2022.10.028. Online ahead of print.ABSTRACTBACKGROUND & AIMS: Acetaminophen (APAP) is the most common cause of drug-induced liver injury (DILI); however, treatment options are limited. Mas is a G protein-coupled receptor whose role in APAP-induced hepatotoxicity has not yet been examined.METHODS: Intrahepatic Mas expression was determined in both human and mouse DILI models. Mas1-/-, AlbcreMas1f/f, Ppara-/-, Mas1-/-Ppara-/- and wild-type mice were challenged with APAP for the in vivo analyses of Mas-AKT-FOXO1 axis-dependent lipophagy and fatty acid oxidation (FAO), using pharmacological compounds and genetic tools. Liver samples were collected for RNA-sequencing, proteomics, metabolomics, lipidomics, and metabolic flux analysis. Live-imaging of liver and histological, biochemical, and molecular studies were performed to evaluate APAP-induced hepatotoxicity in mice. Primary hepatocytes and hepatic cell lines were exposed to APAP for in vitro analysis.RESULTS: Intrahepatic Mas expression was significantly upregulated in human and mouse DILI models. Mice with systemic, liver-specific, or hepatocyte-specific Mas1 deficiency were vulnerable to APAP-induced hepatotoxicity. They exhibited substantially impaired lipophagy and downstream FAO, which was accompanied by the activation of AKT and suppression of FOXO1. In addition, the prophylactic activation of Mas showed unbelievably ideal effects to protect mice from APAP challenge, with remarkably enhanced lipophagy and FAO dependent on the AKT-FOXO1 axis. Moreover, the protective effects of AVE0991 were substantially diminished by the inhibition of either lipophagy or FAO.CONCLUSIONS: The activation of Mas on hepatocytes enhanced AKT-FOXO1-dependent lipophagy and downstream FAO to protect mice from APAP-induced hepatotoxicity, suggesting that hepatocyte-specific Mas might be a novel therapeutic target for DILI.IMPACT AND IMPLICATIONS: Mas signaling arises as a novel therapeutic target for patients with APAP overdose. Mas-AKT/FOXO1-fatty acid degradation pathway is critical for researchers to develop treatment strategies of APAP overdose. When Mas signaling is targeted, the extent of liver injury should be taken into account at the time of administration. These findings obtained from APAP-challenged mice still need to be confirmed in the clinics.PMID:36368597 | DOI:10.1016/j.jhep.2022.10.028

Gene. 2022 Nov 8:147039. doi: 10.1016/j.gene.2022.147039. Online ahead of print.ABSTRACTThe prevalence of non-alcoholic fatty liver diseases (NAFLD) has reached epidemic levels during recent years and a major driver of NAFLD are diets high in fat and fructose. A common practice in the treatment of NAFLD are life-style interventions including for example increased physical activity. The transcriptional coactivator peroxisome proliferator-activated receptor gamma coactivator 1α (PGC-1α) has been shown to be central in mediating the beneficial effects of exercise training by regulating the expression of key metabolic genes. However, the significance of hepatic PGC-1α for the high fat high fructose (HFFD) induced condition of NAFLD has not been elucidated. Therefore the aim of the present study was to investigate the effect of hepatic PGC-1α on HFFD and exercise-induced changes in the hepatic transcriptome and metabolome in mice. Using gene-arrays and 1H NMR spectroscopy, the liver transcriptome and metabolome of liver-specific PGC-1α knock-out mice receiving either standard chow, HFFD or HFFD + exercise (HFFD+Ex) were determined. In total 122 genes were identified as differently expressed in mice receiving HFFD for 13 weeks compared to chow, while the loss of hepatic PGC-1α only had very minor effects on the transcriptome. The same was observed for the liver metabolome. The effect of 4 weeks exercise training in combination with 13 weeks of HFFD, had small effects on the transcriptome compared to HFFD alone, while more effects were observed for the metabolome. Together our results highlights a minor regulatory effect of hepatic PGC-1α on the liver transcriptome during high fat high fructose diet and exercise training.PMID:36368573 | DOI:10.1016/j.gene.2022.147039