Integrative Molecular Phenotyping
INTEGRATIVE MOLECULAR
PHENOTYPING
WHEELOCK LABORATORY
DEPARTMENT OF MEDICAL
BIOCHEMISTRY AND BIOPHYSICS
WHEELOCK LABORATORY
DEPARTMENT OF MEDICAL
BIOCHEMISTRY AND BIOPHYSICS
WHEELOCK LABORATORY
DEPARTMENT OF MEDICAL
BIOCHEMISTRY AND BIOPHYSICS
WHEELOCK LABORATORY
DEPARTMENT OF MEDICAL
BIOCHEMISTRY AND BIOPHYSICS
WHEELOCK LABORATORY
DEPARTMENT OF MEDICAL
BIOCHEMISTRY AND BIOPHYSICS
WHEELOCK LABORATORY

PubMed

Conjoint analysis of transcriptome and metabolome profiles of normal captivity and arch soil free-range in Meishan pigs

Mon, 14/08/2023 - 12:00
Front Vet Sci. 2023 Jul 27;10:1187877. doi: 10.3389/fvets.2023.1187877. eCollection 2023.ABSTRACTThe hygiene hypothesis has been advanced as a potential explanation for the increasingly high levels of atopy and allergic disease in the general human population. In an effort to conduct a more detailed study of the link between immune activity and the hygiene hypothesis, Meishan pigs raised under normal captivity (NC) or arch soil free-range (ASF) conditions were selected as an experimental model system. Cytokine levels were found to differ significantly between these two groups consistent with a difference in cellular immune status. Integrated transcriptomic and metabolomic profiling of duodenal tissue samples from Meishan pigs were then performed, leading to the identification of differentially expressed genes (DEGs), differentially abundant metabolites (DAMs), and key pathways that were able to distinguish the NC and ASF groups. This approach led to the identification of 1,113 DEGs, as well as 577 and 372 DAMs in positive and negative ion modes, respectively. When an interaction network incorporating DEGs and metabolites associated with immune responsivity was constructed, it included factors such as 9-cis-Retinoic acid, (9Z,11E)-(13S)-13-Hydroxyoctadeca-9,11-dienoic acid and (10E,12Z)-(9S)-9-Hydroxyoctadeca-10,12-dienoic acid. Functional enrichment analyses confirmed that identified DEGs and DAMs were associated with immune-related pathways including the intestinal IgA production and PPAR signaling pathways. Together, these results offer new insight into the roles that particular genes and metabolites enriched in response to environmental stressors in free-range Meishan pigs may play in the regulation of cellular immunity, thus offering a foundation for future efforts to better understand the immunological mechanisms underlying the hygiene hypothesis.PMID:37576833 | PMC:PMC10421962 | DOI:10.3389/fvets.2023.1187877

Metabolomic approach for phytochemical assessment of <em>Murraya koenigii</em> fruits during different maturity stages

Mon, 14/08/2023 - 12:00
Heliyon. 2023 Jul 29;9(8):e18688. doi: 10.1016/j.heliyon.2023.e18688. eCollection 2023 Aug.ABSTRACTA detailed metabolomic study was performed on various maturation stages of Murraya koenigii fruit pulps, seed, and leaf. Among the fruit pulps, stage 6 had the highest TPC (13.27 mg/g of GAE) and TFC content (6.16 mg/g RE). The extracts also showed promising free radical scavenging activity, especially in the seed (IC50DPPH 427 μg/mL). Metabolomics study revealed the identification of 133 metabolites in fruit pulps, seeds and leaves using the METLIN database. In silico PASS software analysis predicted the antimutagenic property of myricetin and bismurrayaquinone A. Pathway analysis revealed the phenylpropanoid biosynthesis pathway as one of the major pathways present in the fruit pulps. This detailed metabolic report of M. koenigii fruit maturation report brings a new insight into phytochemicals and their distribution in seed, pulps and leaves along with nutritive values and can be considered for nutritive and therapeutic purposes.PMID:37576304 | PMC:PMC10415817 | DOI:10.1016/j.heliyon.2023.e18688

Metabolomics: A suitable foodomics approach to the geographical origin traceability of Ethiopian Arabica specialty coffees

Mon, 14/08/2023 - 12:00
Food Sci Nutr. 2023 May 24;11(8):4419-4431. doi: 10.1002/fsn3.3434. eCollection 2023 Aug.ABSTRACTCoffee arabica, originated in Ethiopia, is considered a quality bean for its high sensory qualities, and has a special price in the world coffee market. The country is a pool of genetic diversity for Arabica coffee, and coffee from different regions has a distinct flavor profile. Their exceptional quality is attributed to their genetic diversity, favorable environmental conditions, and agroforestry-based production system. However, the country still needs to benefit from its single-origin product due to a lack of appropriate traceability information to register for its geographical indication. Certification of certain plants or plant-derived products emerged to inform consumers about their exceptional qualities due to their geographical origin and protect the product from fraud. The recently emerging foodomics approaches, namely proteomics, genomics, and metabolomics, are reported as suitable means of regional agri-food product authentication and traceability. Particularly, the metabolomics approach provides truthful information on product traceability. Despite efforts by some researchers to trace the geographical origin of Ethiopian Arabica coffees through stable isotope and phenolic compound profiling and elemental analysis, foodomics approaches are not used to trace the geographical origin of Arabica specialty coffees from various parts of the country. A metabolomics-based traceability system that demonstrates the connection between the exceptional attributes of Ethiopian Arabica specialty coffees and their geographic origin is recommended to maximize the benefit of single-origin coffees.PMID:37576063 | PMC:PMC10420859 | DOI:10.1002/fsn3.3434

Effect of short-term consumption of yellow peas as noodles on the intestinal environment: A single-armed pre-post comparative pilot study

Mon, 14/08/2023 - 12:00
Food Sci Nutr. 2023 May 10;11(8):4572-4582. doi: 10.1002/fsn3.3416. eCollection 2023 Aug.ABSTRACTLegumes contain dietary fiber and resistant starch, which are beneficial to the intestinal environment. Here, we investigated the effects of yellow pea noodle consumption on the gut microbiota and fecal metabolome of healthy individuals. This single-armed pre-post comparative pilot study evaluated eight healthy female participants who consumed yellow pea noodles for 4 weeks. The gut microbiota composition and fecal metabolomic profile of each participant were evaluated before (2 weeks), during (4 weeks), and after (4 weeks) daily yellow pea noodle consumption. 16S rRNA gene sequencing was performed on stool samples, followed by clustering of operational taxonomic units using the Cluster Database at High Identity with Tolerance and integrated QIIME pipeline to elucidate the gut microbiota composition. The fecal metabolites were analyzed using capillary electrophoresis time-of-flight mass spectrometry and liquid chromatography time-of-flight mass spectrometry. Compared to day 0, the relative abundances of five bacterial genera (Bacteroides, Bilophila, Hungatella, Parabacteroides, and Streptococcus) in the intestinal microbiota significantly decreased, wherein those of Bifidobacterium longum and Ruminococcus bromii were increased on day 29 and decreased to the basal level (day 0) on day 57. Fecal metabolomic analysis identified 11 compounds showing significant fluctuations in participants on day 29 compared to day 0. Although the average levels of short-chain fatty acids in participants did not differ significantly on day 29 compared to those on day 0, the levels tended to increase in individual participants with >8% relative abundance of R. bromii in their gut microbiota. In conclusion, incorporating yellow peas as a daily staple may confer human health benefits by favorably altering the intestinal environment.PMID:37576055 | PMC:PMC10420782 | DOI:10.1002/fsn3.3416

Discrimination for geographical origin of <em>Panax quinquefolius</em> L. using UPLC Q-Orbitrap MS-based metabolomics approach

Mon, 14/08/2023 - 12:00
Food Sci Nutr. 2023 Jul 10;11(8):4843-4852. doi: 10.1002/fsn3.3461. eCollection 2023 Aug.ABSTRACTAmerican ginseng, Panax quinquefolius L., is an important medicinal plant with multiple pharmacological effects and high nutritional value. American ginseng from different geographical origins varies in quality and price. However, there was no approach for discriminating American ginseng from different geographical origins to date. In this study, a metabolomic method based on the UPLC-Orbitrap fusion platform was established to comprehensively determine and analyze metabolites of American ginseng from America and Canada, Heilongjiang, Jilin, Liaoning, and Shandong provinces in China. A total of 382 metabolites were detected, including 230 saponins, 30 amino acids and derivatives, 27 organic acids and derivatives, 25 lipids, 17 carbohydrates and derivatives, 10 phenols, 8 nucleotides, and derivatives, as well as 35 other metabolites. Metabolite differences between North America and Asia producing areas were more obvious than within Asia. Twenty metabolites, contributed most to the differentiation of producing areas, were identified as potential markers with prediction accuracy higher than 91%. The results provide new insights into the metabolite composition of American ginseng from different origins, which will help discriminate origins and promote quality control of American ginseng.PMID:37576031 | PMC:PMC10420767 | DOI:10.1002/fsn3.3461

Editorial: Integrative analysis approaches for understanding plant metabolism, metabolite, chemodiversity and their respective regulation

Mon, 14/08/2023 - 12:00
Front Plant Sci. 2023 Jul 27;14:1248983. doi: 10.3389/fpls.2023.1248983. eCollection 2023.NO ABSTRACTPMID:37575933 | PMC:PMC10415068 | DOI:10.3389/fpls.2023.1248983

Long non-coding RNA-mediated competing endogenous RNA regulatory network during flower development and color formation in <em>Melastoma candidum</em>

Mon, 14/08/2023 - 12:00
Front Plant Sci. 2023 Jul 27;14:1215044. doi: 10.3389/fpls.2023.1215044. eCollection 2023.ABSTRACTM. candidum, an evergreen shrubby flower known for its superior adaptation ability in South China, has gained increased attention in garden applications. However, scant attention has been paid to its flower development and color formation process at the non-coding RNA level. To fill this gap, we conducted a comprehensive analysis based on long non-coding RNA sequencing (lncRNA-seq), RNA-seq, small RNA sequencing (sRNA-seq), and widely targeted metabolome detection of three different flower developmental stages of M. candidum. After differentially expressed lncRNAs (DElncRNAs), differentially expressed mRNAs (DEmRNAs), differentially expressed microRNAs (DEmiRNAs), and differentially synthesized metabolites (DSmets) analyses between the different flower developmental stages, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) were conducted to identify some key genes and metabolites in flavonoid, flavone, anthocyanin, carotenoid, and alkaloid-related GO terms and biosynthetic pathways. Three direct-acting models, including antisense-acting, cis-acting, and trans-acting between lncRNAs and mRNAs, were detected to illustrate the direct function of lncRNAs on target genes during flower development and color formation. Based on the competitive endogenous RNA (ceRNA) regulatory theory, we constructed a lncRNA-mediated regulatory network composed of DElncRNAs, DEmiRNAs, DEmRNAs, and DSmets to elucidate the indirect role of lncRNAs in the flower development and color formation of M. candidum. By utilizing correlation analyses between DERNAs and DSmets within the ceRNA regulatory network, alongside verification trials of the ceRNA regulatory mechanism, the study successfully illustrated the significance of lncRNAs in flower development and color formation process. This research provides a foundation for improving and regulating flower color at the lncRNA level in M. candidum, and sheds light on the potential applications of non-coding RNA in studies of flower development.PMID:37575929 | PMC:PMC10415103 | DOI:10.3389/fpls.2023.1215044

Systems-level proteomics and metabolomics reveals the diel molecular landscape of diverse kale cultivars

Mon, 14/08/2023 - 12:00
Front Plant Sci. 2023 Jul 28;14:1170448. doi: 10.3389/fpls.2023.1170448. eCollection 2023.ABSTRACTKale is a group of diverse Brassicaceae species that are nutritious leafy greens consumed for their abundance of vitamins and micronutrients. Typified by their curly, serrated and/or wavy leaves, kale varieties have been primarily defined based on their leaf morphology and geographic origin, despite having complex genetic backgrounds. Kale is a very promising crop for vertical farming due to its high nutritional content; however, being a non-model organism, foundational, systems-level analyses of kale are lacking. Previous studies in kale have shown that time-of-day harvesting can affect its nutritional composition. Therefore, to gain a systems-level diel understanding of kale across its wide-ranging and diverse genetic landscape, we selected nine publicly available and commercially grown kale cultivars for growth under near-sunlight LED light conditions ideal for vertical farming. We then analyzed changes in morphology, growth and nutrition using a combination of plant phenotyping, proteomics and metabolomics. As the diel molecular activities of plants drive their daily growth and development, ultimately determining their productivity as a crop, we harvested kale leaf tissue at both end-of-day (ED) and end-of-night (EN) time-points for all molecular analyses. Our results reveal that diel proteome and metabolome signatures divide the selected kale cultivars into two groups defined by their amino acid and sugar content, along with significant proteome differences involving carbon and nitrogen metabolism, mRNA splicing, protein translation and light harvesting. Together, our multi-cultivar, multi-omic analysis provides new insights into the molecular underpinnings of the diel growth and development landscape of kale, advancing our fundamental understanding of this nutritious leafy green super-food for horticulture/vertical farming applications.PMID:37575922 | PMC:PMC10421703 | DOI:10.3389/fpls.2023.1170448

Association of gut microbiota, plasma and fecal metabolite profiles with intellectual development in school-age children

Mon, 14/08/2023 - 12:00
Transl Pediatr. 2023 Jul 31;12(7):1292-1304. doi: 10.21037/tp-22-610. Epub 2023 Jun 29.ABSTRACTBACKGROUND: Little is known about how the gut microbiota and metabolic profiles are related to cognitive outcomes in young children until now. It was hypothesized that the gut microbiota, the plasma and fecal metabolites significantly correlated with intelligence quotient (IQ) in school-age children in current study.METHODS: This cross-sectional study enrolled 452 children aged 6-9 years old. IQ was measured using the Wechsler Intelligence Scale for Children-Fourth Edition. Fecal microbiota, plasma and fecal metabolites were analyzed using 16S rRNA amplicon sequencing and targeted metabolomic technologies, respectively.RESULTS: Restricted maximum likelihood (REML) analyses showed that microbiota composition and fecal metabolites were associated with neither subscale nor full-scale IQ (P: 0.059-0.500). However, plasma metabolites were significantly correlated with the processing speed (P=0.008). In multiple regression analysis after adjusting for confounders and multiple test correction, benzoic acid, azelaic acid, adipic acid, suberic acid and malonic acid selected by the multivariate methods with unbiased variable selection were positively associated with processing speed index (PSI) [Pfalse discovery rate (FDR): 0.006-0.024], whereas pyruvic acid was negatively associated with the PSI and full-scale IQ (PFDR: 0.014-0.030).CONCLUSIONS: In normal school-age children, certain plasma metabolites concentrations but not the gut microbiota composition nor fecal metabolites are correlated with intelligence.PMID:37575906 | PMC:PMC10416130 | DOI:10.21037/tp-22-610

Paclitaxel Induces Neurotoxicity by Disrupting Tricarboxylic Acid Cycle Metabolic Balance in the Mouse Hippocampus

Mon, 14/08/2023 - 12:00
J Toxicol. 2023 Aug 5;2023:5660481. doi: 10.1155/2023/5660481. eCollection 2023.ABSTRACTOBJECTIVE: It is well known that paclitaxel (PTX)-induced neurotoxicity seriously affects the quality of life of patients and is the main reason for reducing the dose of chemotherapy or even stopping chemotherapy. The current data are limited, and further information is required for practice and verification. The aims of this study were to clarify the molecular mechanism underlying PTX-induced neurotoxicity by combining in vivo and in vitro metabolomics studies and provide new targets for the prevention and treatment of PTX-induced neurotoxicity.METHODS: In the in vivo study, a PTX-induced neurotoxicity mouse model was established by intraperitoneal injection of PTX (6 mg/kg every three days) for two consecutive weeks. After verification by water maze tests and HE staining of pathological sections, hippocampal metabolites were measured and the differential metabolites and related metabolic pathways were identified by multivariate statistical analysis. In the in vitro study, we investigated the effects of PTX on mouse hippocampal neuron cells, assessing the concentration and time of administration by MTT assays. After modeling, the relevant metabolites in the TCA cycle were quantified by targeted metabolomics using stable isotope labeling. Finally, the key enzymes of the TCA cycle in tissues and cells were verified by RT-PCR.RESULTS: Administration of PTX to model mice resulted in neurological damage, shown by both water-maze tests and hippocampal tissue sections. Twenty-four metabolites and five associated metabolic pathways were found to differ significantly between the hippocampal tissues of the model and control groups. These included metabolites and pathways related to the TCA cycle and pyruvate metabolism. Metabolomics analysis using stable isotope labeling showed significant changes in metabolites associated with the TCA cycle compared with the control group (P < 0.05). Finally, RT-PCR verified that the expression of key enzymes in the TCA cycle was changed to different degrees in both hippocampal tissues and cells.CONCLUSION: Our results showed that PTX neurotoxicity in hippocampal tissue and neuron cells was associated with inhibition of the TCA cycle. This inhibition leads to brain insufficiency and impaired metabolism, resulting in various neurotoxic symptoms.PMID:37575636 | PMC:PMC10423086 | DOI:10.1155/2023/5660481

Metabolic Responses of Normal Rat Kidneys to a High Salt Intake

Mon, 14/08/2023 - 12:00
Function (Oxf). 2023 Jun 22;4(5):zqad031. doi: 10.1093/function/zqad031. eCollection 2023.ABSTRACTIn this study, novel methods were developed, which allowed continuous (24/7) measurement of arterial blood pressure and renal blood flow in freely moving rats and the intermittent collection of arterial and renal venous blood to estimate kidney metabolic fluxes of O2 and metabolites. Specifically, the study determined the effects of a high salt (HS; 4.0% NaCl) diet upon whole kidney O2 consumption and arterial and renal venous plasma metabolomic profiles of normal Sprague-Dawley rats. A separate group of rats was studied to determine changes in the cortex and outer medulla tissue metabolomic and mRNAseq profiles before and following the switch from a 0.4% to 4.0% NaCl diet. In addition, targeted mRNA expression analysis of cortical segments was performed. Significant changes in the metabolomic and transcriptomic profiles occurred with feeding of the HS diet. A progressive increase of kidney O2 consumption was found despite a reduction in expression of most of the mRNA encoding enzymes of TCA cycle. A novel finding was the increased expression of glycolysis-related genes in Cx and isolated proximal tubular segments in response to an HS diet, consistent with increased release of pyruvate and lactate from the kidney to the renal venous blood. Data suggests that aerobic glycolysis (eg, Warburg effect) may contribute to energy production under these circumstances. The study provides evidence that kidney metabolism responds to an HS diet enabling enhanced energy production while protecting from oxidative stress and injury. Metabolomic and transcriptomic analysis of kidneys of Sprague-Dawley rats fed a high salt diet.PMID:37575482 | PMC:PMC10413938 | DOI:10.1093/function/zqad031

Urine-based Detection of Congenital Portosystemic Shunt in C57BL/6 Mice

Mon, 14/08/2023 - 12:00
Function (Oxf). 2023 Jul 28;4(5):zqad040. doi: 10.1093/function/zqad040. eCollection 2023.ABSTRACTSporadic occurrence of congenital portosystemic shunt (PSS) at a rate of ∼1 out of 10 among C57BL/6 J mice, which are widely used in biomedical research, results in aberrancies in serologic, metabolic, and physiologic parameters. Therefore, mice with PSS should be identified as outliers in research. Accordingly, we sought methods to, reliably and efficiently, identify PSS mice. Serum total bile acids ≥ 40 µm is a bona fide biomarker of PSS in mice but utility of this biomarker is limited by its cost and invasiveness, particularly if large numbers of mice are to be screened. This led us to investigate if assay of urine might serve as a simple, inexpensive, noninvasive means of PSS diagnosis. Metabolome profiling uncovered that Krebs cycle intermediates, that is, citrate, α-ketoglutarate, and fumarate, were strikingly and distinctly elevated in the urine of PSS mice. We leveraged the iron-chelating and pH-lowering properties of such metabolites as the basis for 3 urine-based PSS screening tests: urinary iron-chelation assay, pH strip test, and phenol red assay. Our findings demonstrate the feasibility of using these colorimetric assays, whereby their readout can be assessed by direct observation, to diagnose PSS in an inexpensive, rapid, and noninvasive manner. Application of our urinary PSS screening protocols can aid biomedical research by enabling stratification of PSS mice, which, at present, likely confound numerous ongoing studies.PMID:37575479 | PMC:PMC10413929 | DOI:10.1093/function/zqad040

Functional Analysis of Plant Monosaccharide Transporters Using a Simple Growth Complementation Assay in Yeast

Mon, 14/08/2023 - 12:00
Bio Protoc. 2023 Aug 5;13(15):e4733. doi: 10.21769/BioProtoc.4733. eCollection 2023 Aug 5.ABSTRACTThe study of genes and their products is an essential prerequisite for fundamental research. Characterization can be achieved by analyzing mutants or overexpression lines or by studying the localization and substrate specificities of the resulting proteins. However, functional analysis of specific proteins in complex eukaryotic organisms can be challenging. To overcome this, the use of heterologous systems to express genes and analyze the resulting proteins can save time and effort. Yeast is a preferred heterologous model organism: it is easy to transform, and tools for genomics, engineering, and metabolomics are already available. Here, we describe a well-established and simple method to analyze the activity of plant monosaccharide transporters in the baker's yeast, Saccharomyces cerevisiae, using a simple growth complementation assay. We used the famous hexose-transport-deficient yeast strain EBY.VW4000 to express candidate plant monosaccharide transporters and analyzed their transport activity. This assay does not require any radioactive labeling of substrates and can be easily extended for quantitative analysis using growth curves or by analyzing the transport rates of fluorescent substrates like the glucose analog 2-NBDG. Finally, to further simplify the cloning of potential candidate transporters, we provide level 0 modular cloning (MoClo) modules for efficient and simple Golden Gate cloning. This approach provides a convenient tool for the functional analysis of plant monosaccharide transporters in yeast. Key features Comprehensive, simple protocol for analysis of plant monosaccharide transporters in yeast Includes optional MoClo parts for cloning with Golden Gate method Includes protocol for the production and transformation of competent yeast cells Does not require hazardous solutions, radiolabeled substrates, or specialized equipment.PMID:37575400 | PMC:PMC10415198 | DOI:10.21769/BioProtoc.4733

Current advances of liquid biopsies in prostate cancer: Molecular biomarkers

Mon, 14/08/2023 - 12:00
Mol Ther Oncolytics. 2023 Jul 19;30:27-38. doi: 10.1016/j.omto.2023.07.004. eCollection 2023 Sep 21.ABSTRACTProstate cancer (PCa) incidence is increasing and endangers men's lives. Early detection of PCa could improve overall survival (OS) by preventing metastasis. The prostate-specific antigen (PSA) test is a popular screening method. Several advisory groups, however, warn against using the PSA test due to its high false positive rate, unsupported outcome, and limited benefit. The number of disease-related biopsies performed annually far outweighs the number of diagnoses. Thus, there is an urgent need to develop accurate diagnostic biomarkers to detect PCa and distinguish between aggressive and indolent cancers. Recently, non-coding RNA (ncRNA), circulating tumor DNA (ctDNA)/ctRNA, exosomes, and metabolomic biomarkers in the liquid biopsies (LBs) of patients with PCa showed significant differences and clinical benefits in diagnosis, prognosis, and monitoring response to therapy. The analysis of urinary exosomal ncRNA presented a substantial correlation among Exos-miR-375 downregulation, clinical T stage, and bone metastases of PCa. Furthermore, the expression of miR-532-5p in urine samples was a vital predictive biomarker of PCa progression. Thus, this review focuses on promising molecular and metabolomic biomarkers in LBs from patients with PCa. We thoroughly addressed the most recent clinical findings of LB biomarker use in diagnosing and monitoring PCa in early and advanced stages.PMID:37575217 | PMC:PMC10415624 | DOI:10.1016/j.omto.2023.07.004

Fetal metabolic adaptations to cardiovascular stress in twin-twin transfusion syndrome

Mon, 14/08/2023 - 12:00
iScience. 2023 Jul 20;26(8):107424. doi: 10.1016/j.isci.2023.107424. eCollection 2023 Aug 18.ABSTRACTMonochorionic-diamniotic twin pregnancies are susceptible to unique complications arising from a single placenta shared by two fetuses. Twin-twin transfusion syndrome (TTTS) is a constellation of disturbances caused by unequal blood flow within the shared placenta giving rise to a major hemodynamic imbalance between the twins. Here, we applied TTTS as a model to uncover fetal metabolic adaptations to cardiovascular stress. We compared untargeted metabolomic analyses of amniotic fluid samples from severe TTTS cases vs. singleton controls. Amniotic fluid metabolites demonstrated alterations in fatty acid, glucose, and steroid hormone metabolism in TTTS. Among TTTS cases, unsupervised principal component analysis revealed two distinct clusters of disease defined by levels of glucose metabolites, amino acids, urea, and redox status. Our results suggest that the human fetal heart can adapt to hemodynamic stress by modulating its glucose metabolism and identify potential differences in the ability of individual fetuses to respond to cardiovascular stress.PMID:37575192 | PMC:PMC10415929 | DOI:10.1016/j.isci.2023.107424

<sub>Inducible Nitric Oxide Synthase Activity Mediates TNF-α-Induced Endothelial</sub> <sub>Cell Dysfunction</sub>

Mon, 14/08/2023 - 12:00
Am J Physiol Cell Physiol. 2023 Aug 14. doi: 10.1152/ajpcell.00153.2023. Online ahead of print.ABSTRACTInducible nitric oxide synthase (iNOS) and vascular endothelial dysfunction have been implicated in the development and progression of atherosclerosis. This study aimed to elucidate the role of iNOS in vascular endothelial dysfunction. Ultrahigh performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC/QTOF-MS) combined with multivariate data analysis was used to characterize the metabolic changes in human umbilical vein endothelial cells (HUVECs) in response to different treatment conditions. In addition, molecular biology techniques were employed to explain the molecular mechanisms underlying the role of iNOS in vascular endothelial dysfunction. Tumor necrosis factor-alpha (TNF-α) enhances the expression of iNOS, TXNIP, and the level of reactive oxygen species (ROS), facilitates the entry of nuclear factor-kappa B (NF-κB) into the nucleus; and promotes injury in HUVECs. iNOS deficiency reversed the TNF-α-mediated pathological changes in HUVECs. Moreover, TNF-α increased the expression of TNFR-2, and the levels of p-IκBα and IL-6 proteins, and CD31, ICAM-1, and VCAM-1 proteins expression which were significantly reduced in HUVECs with iNOS deficiency. In addition, treating HUVECs in the absence or presence of TNF-α or iNOS, respectively, enabled the identification of putative endogenous biomarkers associated with endothelial dysfunction. These biomarkers were involved in critical metabolic pathways, including glycosylphosphatidylinositol-anchor biosynthesis, amino acid metabolism, sphingolipid metabolism, and fatty acid metabolism. iNOS deficiency during vascular endothelial dysfunction may affect the expression of TNFR-2, vascular adhesion factors and the level of ROS via cellular metabolic changes, thereby attenuating vascular endothelial dysfunction.PMID:37575057 | DOI:10.1152/ajpcell.00153.2023

H/D Exchange Coupled with <sup>2</sup>H-labeled Stable Isotope-Assisted Metabolomics Discover Transformation Products of Contaminants of Emerging Concern

Mon, 14/08/2023 - 12:00
Anal Chem. 2023 Aug 13. doi: 10.1021/acs.analchem.3c02833. Online ahead of print.ABSTRACTStable isotope-assisted metabolomics (SIAM) is a powerful tool for discovering transformation products (TPs) of contaminants. Nevertheless, the high cost or lack of isotope-labeled analytes limits its application. In-house H/D (hydrogen/deuterium) exchange reactions enable direct 2H labeling to target analytes with favorable reaction conditions, providing intuitive and easy-to-handle approaches for environmentally relevant laboratories to obtain cost-effective 2H-labeled contaminants of emerging concern (CECs). We first combined the use of in-house H/D exchange and 2H-SIAM to discover potential TPs of 6PPD (N-1,3-dimethylbutyl-N'-phenyl-p-phenylenediamine), providing a new strategy for finding TPs of CECs. 6PPD-d9 was obtained by in-house H/D exchange with favorable reaction conditions, and the impurities were carefully studied. Incomplete deuteride, for instance, 6PPD-d8 in this study, constitutes a major part of the impurities. Nevertheless, it has few adverse effects on the 2H-SIAM pipeline in discovering TPs of 6PPD. The 2H-SIAM pipeline annotated 9 TPs of 6PPD, and commercial standards further confirmed the annotated 6PPDQ (2-anilino-5-(4-methylpentan-2-ylamino)cyclohexa-2,5-diene-1,4-dione) and PPPD (N-phenyl-p-phenylenediamine). Additionally, a possible new formation mechanism for 6PPDQ was proposed, highlighting the performance of the strategy. In summary, this study highlighted a new strategy for discovering the TPs of CECs and broadening the application of SIAM in environmental studies.PMID:37574906 | DOI:10.1021/acs.analchem.3c02833

Variations in metabolite profiles of serum coronas produced around PEGylated liposomal drugs by surface property

Mon, 14/08/2023 - 12:00
Colloids Surf B Biointerfaces. 2023 Aug 1;230:113488. doi: 10.1016/j.colsurfb.2023.113488. Online ahead of print.ABSTRACTUnderstanding biomolecular coronas that spontaneously occur around nanocarriers (NCs) in biological fluids is critical to nanomedicine as the coronas influence the behaviors of NCs in biological systems. In contrast to extensive investigations of protein coronas over the past decades, understanding of the coronas of biomolecules beyond proteins, e.g., metabolites, has been rather limited despite such biochemicals being ubiquitously involved in the coronas, which may influence the bio-nano interactions and thus exert certain biological impacts. In this study, serum biomolecular coronas, in particular the coronas of metabolites including lipids, around PEGylated doxorubicin-loaded liposomes with different surface property were investigated. The surface properties of liposomal drugs varied in terms of surface charge and PEGylation density by employing different ionic lipids such as DOTAP and DOPS and different concentrations of PEGylation lipids in liposome formulation. Using the liposomal drugs, the influence of the surface property on the serum metabolite profiles in the coronas was traced for target molecules of 220 lipids and 88 hydrophilic metabolites. From the results, it was found that metabolites rather than proteins mainly constitute the serum coronas on the liposomal drugs. Most of the serum metabolites were found to be retained in the coronas but with altered abundances. Depending on their class, lipids exhibited a different dependence on the surface property. However, overall, lipids appeared to favor corona formation on more negatively charged and PEGylated surfaces. Hydrophilic metabolites also exhibited a similar propensity for corona formation. This study on the surface dependence of metabolite corona formation provides a fundamental contribution toward attaining a comprehensive understanding of biomolecular coronas, which will be critical to the development of efficient nanomedicine.PMID:37574616 | DOI:10.1016/j.colsurfb.2023.113488

Impact of low temperature on the chemical profile of sweet corn kernels during post-harvest storage

Sun, 13/08/2023 - 12:00
Food Chem. 2023 Aug 3;431:137079. doi: 10.1016/j.foodchem.2023.137079. Online ahead of print.ABSTRACTFresh sweet corn has a limited shelf-life due to its high moisture and high sugar content. Low temperature storage is an effective technique employed to extend the shelf-life. However, changes in the chemical composition of sweet corn kernels at low temperatures are not fully understood. In this study, kernels stored at low temperature exhibited higher levels of soluble sugars and lower starch content. In total, 1365 metabolites were characterized in sweet corn kernels. 593 and 308 differentially accumulated metabolites (DAMs) were identified in sweet corn kernels stored at normal and low temperature, respectively. 607 DAMs were identified at low temperature compare to normal temperature. DAMs were consistently enriched in flavonoid biosynthesis, linoleic acid metabolism and sphingolipid metabolism. Moreover, dozens of metabolites were identified as potential biomarkers for post-harvest storage effects in sweet corn. These results extend our knowledge of the dynamic changes in sweet corn kernels stored at low temperatures.PMID:37573745 | DOI:10.1016/j.foodchem.2023.137079

In-depth mass spectrometry analysis of rhGH administration altered energy metabolism and steroidogenesis

Sun, 13/08/2023 - 12:00
Talanta. 2023 Aug 9;266(Pt 2):125069. doi: 10.1016/j.talanta.2023.125069. Online ahead of print.ABSTRACTGrowth hormone, as a proteohormone, is primarily known of its dramatic effect on longitudinal growth. Recombinant DNA technology has provided a safe, abundant and comparatively cheap supply of human GH for growth hormone-deficient individuals. However, many healthy subjects, especially athletics, administrate GH for enhanced athletic performance or strength. A better and more comprehensive understanding of rhGH effect in healthy individuals is urgent and essential. In this study, we recruited 14 healthy young male and injected rhGH once. Untargeted LC-MS metabolomics profiling of serum and urine was performed before and after the rhGH injection. The GH-induced dysregulation of energy related pathways, such as amino acid metabolism, nucleotide metabolism, glycolysis and TCA cycle, was revealed. Moreover, individuals supplemented with micro-doses of rhGH exhibited significantly changed urinary steroidal profiles, suggesting a role of rhGH in both energy metabolism and steroidogenesis. We expect that our results will be helpful to provide new evidence on the effects of rhGH injection and provide potential biomarkers for rhGH administration.PMID:37574608 | DOI:10.1016/j.talanta.2023.125069

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