PubMed

Int J Bioprint. 2023 Feb 21;9(5):691. doi: 10.18063/ijb.691. eCollection 2023.ABSTRACTEdible bird's nests (EBN)-the nests of swiftlet birds harvested from the wild- are high-end healthcare food in East Asia, while their excessive harvesting poses increasing ecological, environmental, and food safety concerns. Here, we report for the first time a tissue-engineering (TE) approach for fabricating EBNs substitutes by integrating the technologies of three-dimensional (3D) printing and live cell culture. The engineered products, tissue-engineered edible bird's nests (TeeBN), comprise two layers. The first is a feeding layer that encapsulates epithelial cells in 3D-printed biocompatible gelation scaffolds. These cells secrete bioactive ingredients, e.g., sialic acid and epidermal growth factors (EGF), recapitulating the natural production of these substances by birds. The second is a receiving layer, consisting of foodgrade natural polymers, e.g., polysaccharides, which mimics the building blocks of natural EBNs while biologically stabilizing the factors released from the feeding layer. In vitro characterizations demonstrate that the feeding layer facilitates 3D cell growth and functions, and the receiving layer (as the end product) contains the necessary nutrients expected from natural EBNs-while without harmful substances commonly detected in natural EBNs. Further, in vivo metabolomics studies in mice indicate that TeeBN showed a similar profile of serum metabolites as natural EBN, reflecting comparable nutritional effects. In summary, we innovatively developed a tissue engineering-based substitute for EBNs with comparable metabolic functions and minimized safety risks, opening a new avenue for producing delicacy food from laboratorial cell culture with 3D printing technology.PMID:37457942 | PMC:PMC10339468 | DOI:10.18063/ijb.691

J Endocr Soc. 2023 Jul 1;7(8):bvad091. doi: 10.1210/jendso/bvad091. eCollection 2023 Jul 3.ABSTRACTCONTEXT: Obesity surveillance is scarce in adolescents, and little is known on whether salivary metabolomics data, emerging minimally invasive biomarkers, can characterize metabolic patterns associated with overweight or obesity in adolescents.OBJECTIVE: This pilot study aims to identify the salivary molecular signatures associated with body mass index (BMI) in Italian adolescents.METHODS: Saliva samples and BMI were collected in a subset of n = 74 young adolescents enrolled in the Public Health Impact of Metal Exposure study (2007-2014). A total of 217 untargeted metabolites were identified using liquid chromatography-high resolution mass spectrometry. Robust linear regression was used to cross-sectionally determine associations between metabolomic signatures and sex-specific BMI-for-age z-scores (z-BMI).RESULTS: Nearly 35% of the adolescents (median age: 12 years; 51% females) were either obese or overweight. A higher z-BMI was observed in males compared to females (P = .02). One nucleoside (deoxyadenosine) and 2 lipids (18:0-18:2 phosphatidylcholine and dipalmitoyl-phosphoethanolamine) were negatively related to z-BMI (P < .05), whereas 2 benzenoids (3-hydroxyanthranilic acid and a phthalate metabolite) were positively associated with z-BMI (P < .05). In males, several metabolites including deoxyadenosine, as well as deoxycarnitine, hyodeoxycholic acid, N-methylglutamic acid, bisphenol P, and trigonelline were downregulated, while 3 metabolites (3-hydroxyanthranilic acid, theobromine/theophylline/paraxanthine, and alanine) were upregulated in relation to z-BMI (P < .05). In females, deoxyadenosine and dipalmitoyl-phosphoethanolamine were negatively associated with z-BMI while deoxycarnitine and a phthalate metabolite were positively associated (P < .05). A single energy-related pathway was enriched in the identified associations in females (carnitine synthesis, P = .04).CONCLUSION: Salivary metabolites involved in nucleotide, lipid, and energy metabolism were primarily altered in relation to BMI in adolescents.PMID:37457847 | PMC:PMC10341611 | DOI:10.1210/jendso/bvad091

Front Mol Biosci. 2023 Jun 15;10:1182643. doi: 10.3389/fmolb.2023.1182643. eCollection 2023.ABSTRACTEmerging evidence reveals the fundamental role of the gut microbiome in human health. Among various factors regulating our gut microbiome, diet is one of the most indispensable and prominent one. Inulin is one of the most widely-studied dietary fiber for its beneficial prebiotic effects by positively modulating the gut microbiome and microbial metabolites. Recent research underscores sexual dimorphism and sex-specific disparities in microbiome and also diet-microbiome interactions. However, whether and how the prebiotic effects of dietary fiber differ among sexes remain underexplored. To this end, we herein examine sex-specific differences in the prebiotic effects of inulin on gut microbiome and metabolome in a humanized murine model of aging i.e., aged mice carrying human fecal microbiota. The findings demonstrate that inulin exerts prebiotic effects, but in a sex-dependent manner. Overall, inulin increases the proportion of Bacteroides, Blautia, and glycine, while decreasing Eggerthella, Lactococcus, Streptococcus, trimethylamine, 3-hydroxyisobutyrate, leucine and methionine in both sexes. However, we note sex-specific effects of inulin including suppression of f_Enteroccaceae:_, Odoribacter, bile acids, malonate, thymine, valine, acetoin, and ethanol while promotion of Dubosiella, pyruvate, and glycine in males. Whereas, suppression of Faecalibaculum, Lachnoclostridium, Schaedlerella, phenylalanine and enhancement of Parasutterella, Phocaeicola, f_Lachnospiraceae;_, Barnesiella, Butyricimonas, glycine, propionate, acetate and glutamate are observed in females. Altogether, the study reveals that prebiotic mechanisms of dietary fiber vary in a sex-dependent manner, underscoring the importance of including both sexes in preclinical/clinical studies to comprehend the mechanisms and functional aspects of dietary interventions for effective extrapolation and translation in precision nutrition milieus.PMID:37457834 | PMC:PMC10345844 | DOI:10.3389/fmolb.2023.1182643

Front Mol Biosci. 2023 Jun 29;10:1204273. doi: 10.3389/fmolb.2023.1204273. eCollection 2023.ABSTRACTHow the human body reacts to the exposure of HIV-1 is an important research goal. Frequently, HIV exposure leads to infection, but some individuals show natural resistance to this infection; they are known as HIV-1-exposed but seronegative (HESN). Others, although infected but without antiretroviral therapy, control HIV-1 replication and progression to AIDS; they are named controllers, maintaining low viral levels and an adequate count of CD4+ T lymphocytes. Biological mechanisms explaining these phenomena are not precise. In this context, metabolomics emerges as a method to find metabolites in response to pathophysiological stimuli, which can help to establish mechanisms of natural resistance to HIV-1 infection and its progression. We conducted a cross-sectional study including 30 HESN, 14 HIV-1 progressors, 14 controllers and 30 healthy controls. Plasma samples (directly and deproteinized) were analyzed through Nuclear Magnetic Resonance (NMR) metabolomics to find biomarkers and altered metabolic pathways. The metabolic profile analysis of progressors, controllers and HESN demonstrated significant differences with healthy controls when a discriminant analysis (PLS-DA) was applied. In the discriminant models, 13 metabolites associated with HESN, 14 with progressors and 12 with controllers were identified, which presented statistically significant mean differences with healthy controls. In progressors, the metabolites were related to high energy expenditure (creatinine), mood disorders (tyrosine) and immune activation (lipoproteins), phenomena typical of the natural course of the infection. In controllers, they were related to an inflammation-modulating profile (glutamate and pyruvate) and a better adaptive immune system response (acetate) associated with resistance to progression. In the HESN group, with anti-inflammatory (lactate and phosphocholine) and virucidal (lactate) effects which constitute a protective profile in the sexual transmission of HIV. Concerning the significant metabolites of each group, we identified 24 genes involved in HIV-1 replication or virus proteins that were all altered in progressors but only partially in controllers and HESN. In summary, our results indicate that exposure to HIV-1 in HESN, as well as infection in progressors and controllers, affects the metabolism of individuals and that this affectation can be determined using NMR metabolomics.PMID:37457832 | PMC:PMC10339029 | DOI:10.3389/fmolb.2023.1204273

Food Chem (Oxf). 2023 Jul 3;7:100176. doi: 10.1016/j.fochms.2023.100176. eCollection 2023 Dec 30.ABSTRACTAn integrated analysis of the transcriptome and metabolome was conducted to investigate the underlying mechanisms of apple fruit response to impact damage stress. During the post-damage storage, a total of 124 differentially expressed genes (DEGs) were identified, which were mainly annotated in 13 pathways, including phenylpropanoid biosynthesis. Besides, 175 differentially expressed metabolites (DEMs), including 142 up-regulated and 33 down-regulated metabolites, exhibited significant alteration after impact damage. The DEGs and DEMs were simultaneously annotated in 7 metabolic pathways, including flavonoid biosynthesis. Key genes in the volatile esters and flavonoid biosynthesis pathways were revealed, which may play a crucial role in the coping mechanisms of apple fruit under impact damage stress. Moreover, 13 ABC transporters were significantly upregulated, indicating that ABC transporters may contribute to the transportation of secondary metabolites associated with response to impact damage stress. The results may elucidate the comprehension of metabolic networks and molecular mechanisms in apple fruits that have undergone impact damage.PMID:37457816 | PMC:PMC10344661 | DOI:10.1016/j.fochms.2023.100176

Front Immunol. 2023 Jun 29;14:1158455. doi: 10.3389/fimmu.2023.1158455. eCollection 2023.ABSTRACTINTRODUCTION: Severe COVID-19 results initially in pulmonary infection and inflammation. Symptoms can persist beyond the period of acute infection, and patients with Post-Acute Sequelae of COVID (PASC) often exhibit a variety of symptoms weeks or months following acute phase resolution including continued pulmonary dysfunction, fatigue, and neurocognitive abnormalities. We hypothesized that dysregulated NAD metabolism contributes to these abnormalities.METHODS: RNAsequencing of lungs from transgenic mice expressing human ACE2 (K18-hACE2) challenged with SARS-CoV-2 revealed upregulation of NAD biosynthetic enzymes, including NAPRT1, NMNAT1, NAMPT, and IDO1 6 days post-infection.RESULTS: Our data also demonstrate increased gene expression of NAD consuming enzymes: PARP 9,10,14 and CD38. At the same time, SIRT1, a protein deacetylase (requiring NAD as a cofactor and involved in control of inflammation) is downregulated. We confirmed our findings by mining sequencing data from lungs of patients that died from SARS-CoV-2 infection. Our validated findings demonstrating increased NAD turnover in SARS-CoV-2 infection suggested that modulating NAD pathways may alter disease progression and may offer therapeutic benefits. Specifically, we hypothesized that treating K18-hACE2 mice with nicotinamide riboside (NR), a potent NAD precursor, may mitigate lethality and improve recovery from SARS-CoV-2 infection. We also tested the therapeutic potential of an anti- monomeric NAMPT antibody using the same infection model. Treatment with high dose anti-NAMPT antibody resulted in significantly decreased body weight compared to control, which was mitigated by combining HD anti-NAMPT antibody with NR. We observed a significant increase in lipid metabolites, including eicosadienoic acid, oleic acid, and palmitoyl carnitine in the low dose antibody + NR group. We also observed significantly increased nicotinamide related metabolites in NR treated animals.DISCUSSION: Our data suggest that infection perturbs NAD pathways, identify novel mechanisms that may explain some pathophysiology of CoVID-19 and suggest novel strategies for both treatment and prevention.PMID:37457744 | PMC:PMC10344451 | DOI:10.3389/fimmu.2023.1158455

Front Plant Sci. 2023 Jun 29;14:1201486. doi: 10.3389/fpls.2023.1201486. eCollection 2023.ABSTRACTPogostemon cablin is a well-known protected species widely used in medicine and spices, however the underlying molecular mechanisms and metabolite dynamics of P. cablin flower development remain unclear due to the difficulty in achieving flowering in this species. A comparison of the transcriptome and widely targeted metabolome during P. cablin flower development was first performed in this study. Results showed that a total of 13,469 differentially expressed unigenes (DEGs) and 371 differentially accumulated metabolites (DAMs) were identified. Transcriptomic analysis revealed that the DEGs were associated with starch and sucrose metabolism, terpenoid biosynthesis and phenylpropanoid biosynthesis. Among these DEGs, 75 MIKC-MADS unigenes were associated with the development of floral organs. Gibberellins (GAs), auxin, and aging signaling might form a cross-regulatory network to regulate flower development in P. cablin. According to the metabolic profile, the predominant DAMs were amino acids, flavonoids, terpenes, phenols, and their derivatives. The accumulation patterns of these predominant DAMs were closely associated with the flower developmental stage. The integration analysis of DEGs and DAMs indicated that phenylpropanoids, flavonoids, and amino acids might be accumulated due to the activation of starch and sucrose metabolism. Our results provide some important insights for elucidating the reproductive process, floral organ, and color formation of P. cablin flowers at the molecular level. These results will improve our understanding of the molecular and genetic mechanisms involved in the floral development of P. cablin.PMID:37457333 | PMC:PMC10340533 | DOI:10.3389/fpls.2023.1201486

Front Public Health. 2023 Jun 28;11:1188673. doi: 10.3389/fpubh.2023.1188673. eCollection 2023.NO ABSTRACTPMID:37457250 | PMC:PMC10338829 | DOI:10.3389/fpubh.2023.1188673

Front Physiol. 2023 Jun 30;14:1082953. doi: 10.3389/fphys.2023.1082953. eCollection 2023.ABSTRACTAltered mito-ribosomal fidelity is an important and insufficiently understood causative agent of mitochondrial dysfunction. Its pathogenic effects are particularly well-known in the case of mitochondrially induced deafness, due to the existence of the, so called, ototoxic variants at positions 847C (m.1494C) and 908A (m.1555A) of 12S mitochondrial (mt-) rRNA. It was shown long ago that the deleterious effects of these variants could remain dormant until an external stimulus triggered their pathogenicity. Yet, the link from the fidelity defect at the mito-ribosomal level to its phenotypic manifestation remained obscure. Recent work with fidelity-impaired mito-ribosomes, carrying error-prone and hyper-accurate mutations in mito-ribosomal proteins, have started to reveal the complexities of the phenotypic manifestation of mito-ribosomal fidelity defects, leading to a new understanding of mtDNA disease. While much needs to be done to arrive to a clear picture of how defects at the level of mito-ribosomal translation eventually result in the complex patterns of disease observed in patients, the current evidence indicates that altered mito-ribosome function, even at very low levels, may become highly pathogenic. The aims of this review are three-fold. First, we compare the molecular details associated with mito-ribosomal fidelity to those of general ribosomal fidelity. Second, we gather information on the cellular and organismal phenotypes associated with defective translational fidelity in order to provide the necessary grounds for an understanding of the phenotypic manifestation of defective mito-ribosomal fidelity. Finally, the results of recent experiments directly tackling mito-ribosomal fidelity are reviewed and future paths of investigation are discussed.PMID:37457031 | PMC:PMC10349377 | DOI:10.3389/fphys.2023.1082953

iScience. 2023 Jun 14;26(7):107127. doi: 10.1016/j.isci.2023.107127. eCollection 2023 Jul 21.ABSTRACTNon-alcoholic fatty liver disease (NAFLD) is a highly prevalent disease with no specific drug therapy. High-throughput metabolomics present an unprecedented opportunity to identify biomarkers and potentially causal risk factors for NAFLD. Here, we determined the impact of 21 circulating metabolites, 17 lipids, and 132 lipoprotein particle characteristics on NAFLD combining prospective observational and two-sample Mendelian randomization (MR) analyses in 121,032 UK Biobank participants. We identified several metabolic factors associated with NAFLD risk in observational and MR analyses including triglyceride-rich and high-density lipoprotein particles composition, as well as the ratio of polyunsaturated fatty acids to total fatty acids. This study, is one of the largest to investigate incident NAFLD, provides concordant observational and genetic evidence that therapies aimed at reducing circulating triglycerides and increasing large HDL particles, as well as interventions aimed at increasing polyunsaturated fatty acid content may warrant further investigation into NAFLD prevention and treatment.PMID:37456853 | PMC:PMC10339047 | DOI:10.1016/j.isci.2023.107127

Front Oncol. 2023 Jun 30;13:1142838. doi: 10.3389/fonc.2023.1142838. eCollection 2023.ABSTRACTPancreatic ductal adenocarcinoma (PDAC) is the most common exocrine tumor of the pancreas characterized by late diagnosis, adverse overall 5-year survival, a higher propensity for metastatic disease, and lack of efficacy of systemic therapy options. These adverse outcomes can be partly attributed to complex tumor microenvironment (TME). Over the past decade, immunotherapy has revolutionized the management of certain cancers; thus far, the immunologically 'non-inflamed' tumor microenvironment in PDACs has proven to be challenging. Indolamine 2,3-dioxygenase 1 (IDO1) is the rate-limiting enzyme in the catabolic pathway of L-Tryptophan, an essential amino acid, that gives rise to the immunosuppressive metabolite Kynurenine. IDO1, Indolamine 2,3-dioxygenase 2 (IDO2), and Tryptophan 2,3-dioxygenase (TDO) are the key enzymes in the tryptophan catabolic pathway but we focus on the role of the predominant enzyme form IDO1 in this review. Nicotinamide phosphoribosyl transferase (iNAMPT) regulates the intracellular concentration of NAD and is upregulated in the tumor. In light of the potential role of IDO1 as a driver of hostile TME in PDAC and NAD+ as a key coenzyme in anti-tumor immune response, this review urges focus on extensive research and initiation of clinical trials using IDO1 and NAMPT inhibitors in pancreatic cancer in the future.PMID:37456260 | PMC:PMC10348419 | DOI:10.3389/fonc.2023.1142838

Front Microbiol. 2023 Jun 29;14:1195985. doi: 10.3389/fmicb.2023.1195985. eCollection 2023.ABSTRACTMarigold can protect crops against soil-borne diseases. However, the effects of intercropping with marigold on apple rhizosphere soils are not known. In this study, we investigated the metabolite profiles and bacterial community structures in rhizosphere soils of the apple-marigold intercropping system by high-throughput sequencing and soil metabolomics. The results show that intercropping marigold could significantly enhance soil moisture, nitrogen, and enzyme activities compared with clean tillage. The soil metabolite profiles and the soil bacterial community structures in the rhizosphere soils were different between the inter-and mono-cropping systems. Among nine metabolites, carbohydrates were more increased in the intercropping system than in the monocropping system. Pathway enrichment analysis revealed that the greatest differential, in terms of metabolic pathway, was starch and sucrose metabolism. Moreover, intercropping marigold significantly increased the relative abundance of plant growth promoting bacteria in rhizosphere soils, such as Rhizobiales, Pseudomonadales, and Bacillales. These results indicate that marigold intercropping positively affected the apple orchard's soil quality and may provide a new intercropping technique to improve soil fertility in orchards and promote plant growth.PMID:37455738 | PMC:PMC10343436 | DOI:10.3389/fmicb.2023.1195985

Front Microbiol. 2023 Jun 29;14:1217270. doi: 10.3389/fmicb.2023.1217270. eCollection 2023.ABSTRACTINTRODUCTION: The emergence of multidrug-resistant (MDR) Acinetobacter baumannii prompts clinicians to consider treating these infections with polymyxin combination.METHODS: Metabolomic analysis was applied to investigate the synergistic effects of polymyxin-B, amikacin and sulbactam combination therapy against MDR A. baumannii harboring OXA-23 and other drug resistant genes. The drug concentrations tested were based on their clinical breakpoints: polymyxin-B (2 mg/L), amikacin (16 mg/L), polymyxin-B/amikacin (2/16 mg/L), and polymyxin-B/amikacin/sulbactam (2/16/4 mg/L).RESULTS: The triple antibiotic combination significantly disrupted levels of metabolites involved in cell outer membrane structure including fatty acids, glycerophospholipids, nucleotides, amino acids and peptides as early as 15 min after administration. Amikacin and polymyxin-B alone perturbed a large number of metabolites at 15 min and 1 h, respectively, but the changes in metabolites were short-lived lasting for less than 4 h. In contrast, the combination treatment disrupted a large amount of metabolites beyond 4 h. Compared to the double-combination, the addition of sulbactam to polymyxin-B/amikacin combination produce a greater disorder in A. baumannii metabolome that further confer susceptibility of bacteria to the antibiotics.CONCLUSION: The metabolomic analysis identified mechanisms responsible for the synergistic activities of polymyxin-B/amikacin/sulbactam against MDR A. baumannii.PMID:37455727 | PMC:PMC10343439 | DOI:10.3389/fmicb.2023.1217270

J Inherit Metab Dis. 2023 Jul 16. doi: 10.1002/jimd.12657. Online ahead of print.ABSTRACTSuccinic semialdehyde dehydrogenase deficiency (SSADHD) is a rare neurometabolic disorder caused by disruption of the gamma-aminobutyric acid (GABA) pathway. A more detailed understanding of its pathophysiology, beyond the accumulation of GABA and gamma-hydroxybutyric acid (GHB), will increase our understanding of the disease and may support novel therapy development. To this end, we compared biochemical body fluid profiles from SSADHD patients with controls using next-generation metabolic screening (NGMS). Targeted analysis of NGMS data from cerebrospinal fluid (CSF) showed a moderate increase of aspartic acid, glutaric acid, glycolic acid, 4-guanidinobutanoic acid and 2-hydroxyglutaric acid, and prominent elevations of GHB and 4,5-dihydroxyhexanoic acid (4,5-DHHA) in SSADHD samples. Remarkably, the intensities of 4,5-DHHA and GHB showed a significant positive correlation in control CSF, but not in patient CSF. In an established zebrafish epilepsy model, 4,5-DHHA showed increased mobility that may reflect limited epileptogenesis. Using untargeted metabolomics, we identified 12 features in CSF with high biomarker potential. These had comparable increased fold changes as GHB and 4,5-DHHA. For 10 of these features, a similar increase was found in plasma, urine and/or mouse brain tissue for SSADHD compared to controls. One of these was identified as the novel biomarker 4,5-dihydroxyheptanoic acid. The intensities of selected features in plasma and urine of SSADHD patients positively correlated with the clinical severity score of epilepsy and psychiatric symptoms of those patients, and also showed a high mutual correlation. Our findings provide new insights into the (neuro)metabolic disturbances in SSADHD and give leads for further research concerning SSADHD pathophysiology. This article is protected by copyright. All rights reserved.PMID:37455357 | DOI:10.1002/jimd.12657

Anal Chim Acta. 2023 Sep 15;1274:341578. doi: 10.1016/j.aca.2023.341578. Epub 2023 Jun 29.ABSTRACTAlthough interest in characterizing DNA damage by means of DNA adductomics has substantially grown, the field of DNA adductomics is still in its infancy, with room for optimization of methods for sample analysis, data processing and DNA adduct identification. In this context, the first objective of this study was to evaluate the use of hydrophilic interaction (HILIC) vs. reversed phase liquid chromatography (RPLC) coupled to high resolution mass spectrometry (HRMS) and thermal acidic vs. enzymatic hydrolysis of DNA followed by DNA adduct purification and enrichment using solid-phase extraction (SPE) or fraction collection for DNA adductome mapping. The second objective was to assess the use of total ion count (TIC) and median intensity (MedI) normalization compared to QC (quality control), iQC (internal QC) and quality control-based robust locally estimated scatterplot smoothing (LOESS) signal correction (QC-RLSC) normalization for processing of the acquired data. The results demonstrate that HILIC compared to RPLC allowed better modeling of the tentative DNA adductome, particularly in combination with thermal acidic hydrolysis and SPE (more valid models, with an average Q2(Y) and R2(Y) of 0.930 and 0.998, respectively). Regarding the need for data normalization and the management of (limited) system instability and signal drift, QC normalization outperformed TIC, MedI, iQC and LOESS normalization. As such, QC normalization can be put forward as the default data normalization strategy. In case of momentous signal drift and/or batch effects however, comparison to other normalization strategies (like e.g. LOESS) is recommended. In future work, further optimization of DNA adductomics may be achieved by merging of HILIC and RPLC datasets and/or application of 2D-LC, as well as the inclusion of Schiff base stabilization and/or fraction collection in the thermal acidic hydrolysis-SPE sample preparation workflow.PMID:37455087 | DOI:10.1016/j.aca.2023.341578

Anal Chim Acta. 2023 Sep 15;1274:341574. doi: 10.1016/j.aca.2023.341574. Epub 2023 Jun 29.ABSTRACTBACKGROUND: Gonadotropins are a class of heavily glycosylated protein hormones, thus extremely challenging to characterize by mass spectrometry. As biopharmaceuticals, gonadotropins are prescribed for the treatment of infertility and are derived from different sources: either from pooled urine of pregnant women or upon production in genetically modified Chinese Hamster Ovary cells. Human chorionic gonadotropin (hCG) is sold as a biopharmaceutical under the name Pregnyl® (urinary hCG, u-hCG) and Ovitrelle® (recombinant hCG, r-hCG), and recombinant human follicle stimulating hormone (r-hFSH) is marketed as Gonal-f®. Recently, we reported the exhaustive characterization of r-hCG at different structural levels.RESULTS: We implement size exclusion (SE) HPLC-MS to automatize the acquisition of native mass spectra of r-hCG dimer, but also u-hCG and r-hFSH, comparing the drug products up to intact heterodimer level. A hybrid HPLC-MS approach was employed for the characterization of r-hCG, u-hCG and r-hFSH drug products at different structural levels. Released glycans were analyzed by porous graphitized carbon (PGC)-HPLC-MS/MS, glycopeptides by reversed-phase (RP)-HPLC-MS/MS, subunits by RP-HPLC-MS and finally the intact native heterodimers by semi-automated online buffer exchange SE-HPLC-MS. The data were integrated using bioinformatic tools, to finally unravel the composition of 1481 co-existing dimeric glycoforms for r-hCG, 1167 glycoforms for u-hCG, and 1440 glycoforms for r-hFSH, and to compare critical quality attributes of the different drug products such as their degree of sialylation and O-glycosylation.SIGNIFICANCE AND NOVELTY: The strong alliance of bioanalytics and bioinformatics data integration at the different structural levels allowed the identification of more than thousand different glycoforms of r-hCG, u-hCG, and r-hFSH. The results showed that these biopharmaceuticals differ considerably in their glycosylation patterns and highlight the importance of in-depth characterization of biopharmaceuticals for quality control. © 2017 Elsevier Inc. All rights reserved.PMID:37455084 | DOI:10.1016/j.aca.2023.341574

Anal Chim Acta. 2023 Sep 15;1274:341570. doi: 10.1016/j.aca.2023.341570. Epub 2023 Jun 28.ABSTRACTDipeptides (DPs) have attracted more and more attention in many research fields due to their important biological functions and promising roles as disease biomarkers. However, the determination of DPs in biological samples is very challenging owing to the limited availability of commercial standards, high structure diversity, distinct physical and chemical characteristics, wide concentration range, and the extensive existence of isomers. In this study, a pseudotargeted liquid chromatography-tandem mass spectrometry (LC-MS/MS) method coupled with chemical derivatization for the simultaneous analysis of 400 DPs and their constructing amino acids (AAs) in biospecimens is established. Dansyl chloride (Dns-Cl) chemical derivatization was introduced to provide characteristic MS fragments for annotation and improve the chromatographic separation of DP isomers. A retention time (RT) prediction model was constructed using 83 standards (63 DPs and 20 AAs) based on their quantitative structural retention relationship (QSRR) after the Dns-Cl labeling, which largely facilitated the annotation of the DPs without standards. Finally, we applied this method to investigate the profile change of DPs in a cisplatin-induced acute kidney injury (AKI) rat model. The established workflow provides a platform to profile DPs and expand our understanding of these little-studied metabolites.PMID:37455081 | DOI:10.1016/j.aca.2023.341570

Sci Total Environ. 2023 Jul 14:165499. doi: 10.1016/j.scitotenv.2023.165499. Online ahead of print.ABSTRACTThe consumption of rice contaminated with arsenic on a long-term basis has emerged as a pressing public health issue of global significance. Arsenic-induced urinary injury, particularly kidney damage, has received widespread attention. In this study, mice model under long-term arsenic exposure was established, mouse were exposed to rice arsenic (30 mg/kg) for 14 months. Changes of related metabolites were observed based on kidney metabolomics and lipidomics, and major biomarkers were screened by urine metabolomics. The results showed that phosphatidylethanolamine (PE) was significantly increased and phosphatidycholine (PC) and phosphatidylglycerol (PG) were significantly reduced after arsenic exposure, leading to related downstream lipid metabolism disorders. The metabolic pathways for amino acid and energy were observed to be impacted. In addition, metabolic disorders due to arsenic exposure may be associated with inherited neurometabolic disorders, such as D-2-hydroxyglutaric aciduria (D-2-HGA), and pyruvate carboxylase deficiency (PCD), which is predicted based on significant difference biomarkers (2-oxoglutarate, malic acid, and succinic acid) screened for urine. This study elucidates the mechanism of toxicity in the urinary system induced by arsenic exposure at nearly half life cycle, which furnishes crucial scientific evidence pertaining to the toxicity and risk evaluation associated with chronic exposure to the arsenic.PMID:37454865 | DOI:10.1016/j.scitotenv.2023.165499

Am J Clin Nutr. 2023 Jul 14:S0002-9165(23)66027-7. doi: 10.1016/j.ajcnut.2023.07.006. Online ahead of print.ABSTRACTBACKGROUND: Long-chain polyunsaturated fatty acids (LCPUFA) and their metabolites are closely related to neovascular eye diseases. However, the clinical significance of their oxylipins in retinal vein occlusion (RVO) remains inconclusive.OBJECTIVE: This case-control study aimed to explore metabolomic profiles of LCPUFA oxidation in RVO, and to identify potential indicators for diagnosis and pathological progression.METHODS: The plasma levels of ω-3 and ω-6 LCPUFA and their oxylipins in 44 adults with RVO and 36 normal controls were analyzed using ultra-performance liquid chromatography tandem mass spectrometry. Univariate analysis combined with principal component and orthogonal projections to latent structure discriminant analysis was used to screen differential metabolites. Aortic ring and choroidal explant sprouting assays were used to investigate the effects of 5-oxo-eicosatetraenoic acids (ETE) on angiogenesis ex vivo. Tubule formation and wound healing assays were performed to verify its effects on human retinal microvascular endothelial cell (HRMEC) functions.RESULTS: Higher ω-6 and lower ω-3 LCPUFA plasma levels were measured in the adults with RVO vs. control (OR: 2.34, 95%CI [1.42, 3.86], P < 0.001; OR: 0.28, 95%CI [0.15, 0.51], P < 0.001). Metabolomic analysis revealed 20 LCPUFA and their oxylipins dysregulated in RVO, including increased arachidonic acid (ω-6, OR: 1.85, 95%CI [1.18, 2.90], P < 0.001) and its lipoxygenase product 5-oxo-ETE (OR: 11.76, 95%CI [3.73, 37.11], P < 0.001), as well as decreased docosahexaenoic acid (ω-3, OR: 0.13, 95%CI [0.05, 0.33], P < 0.001). Interestingly, 5-oxo-ETE were downregulated in ischemic vs. nonischemic central RVO. Exogenous 5-oxo-ETE attenuated aortic ring and choroidal explant sprouting, and inhibited tubule formation and migration of HRMECs in a dose-dependent manner, possibly via suppressing the vascular endothelial growth factor signaling pathway.CONCLUSIONS: The plasma levels of ω-6 and ω-3 LCPUFA and their oxylipins were associated with RVO. The ω-6 LCPUFA-derived metabolite 5-oxo-ETE was a potential marker of RVO development and progression.PMID:37454758 | DOI:10.1016/j.ajcnut.2023.07.006

Prog Neuropsychopharmacol Biol Psychiatry. 2023 Jul 14:110830. doi: 10.1016/j.pnpbp.2023.110830. Online ahead of print.ABSTRACTAlzheimer's disease (AD) is often not recognized or is diagnosed very late, which significantly reduces the effectiveness of available pharmacological treatments. Metabolomic analyzes have great potential for improving existing knowledge about the pathogenesis and etiology of AD and represent a novel approach towards discovering biomarkers that could be used for diagnosis, prognosis, and therapy monitoring. In this study, we applied the untargeted metabolomic approach to investigate the changes in biochemical pathways related to AD pathology. We used gas chromatography and liquid chromatography coupled to mass spectrometry (GC-MS and LC-MS, respectively) to identify metabolites whose levels have changed in subjects with AD diagnosis (N = 40) compared to healthy controls (N = 40) and individuals with mild cognitive impairment (MCI, N = 40). The GC-MS identified significant differences between groups in levels of metabolites belonging to the classes of benzene and substituted derivatives, carboxylic acids and derivatives, fatty acyls, hydroxy acids and derivatives, keto acids and derivatives, and organooxygen compounds. Most of the compounds identified by the LC-MS were various fatty acyls, glycerolipids and glycerophospholipids. All of these compounds were decreased in AD patients and in subjects with MCI compared to healthy controls. The results of the study indicate disturbed metabolism of lipids and amino acids and an imbalance of metabolites involved in energy metabolism in individuals diagnosed with AD, compared to healthy controls and MCI subjects.PMID:37454721 | DOI:10.1016/j.pnpbp.2023.110830