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

Analysis of the gut microbiota in children with gastroesophageal reflux disease using metagenomics and metabolomics

Mon, 30/10/2023 - 11:00
Front Cell Infect Microbiol. 2023 Oct 13;13:1267192. doi: 10.3389/fcimb.2023.1267192. eCollection 2023.ABSTRACTBACKGROUND: There is no direct evidence of gut microbiota disturbance in children with gastroesophageal reflux disease (GERD). This study aimed to provide direct evidence and a comprehensive understanding of gut microbiota disturbance in children with GERD through combined metagenomic and metabolomic analysis.METHODS: 30 children with GERD and 30 healthy controls (HCs) were continuously enrolled, and the demographic and clinical characteristics of the subjects were collected. First, 16S rRNA sequencing was used to evaluate differences in the gut microbiota between children with GERD and HC group, and 10 children with GERD and 10 children in the HC group were selected for metagenomic analysis. Nontargeted metabolomic analysis was performed using liquid chromatography/mass spectrometry (LC/MS), and metagenomic and metabolomic data were analyzed together.RESULTS: There were significant differences in the gut microbiota diversity and composition between children with GERD and HCs. The dominant bacteria in children with GERD were Proteobacteria and Bacteroidota. At the species level, the top three core bacterial groups were Bacteroides stercoris, Bacteroides vulgatus and Alistipes putredinis. The main differential pathways were identified to be related to energy, amino acid, vitamin, carbohydrate and lipid metabolism. LC/MS detected 288 different metabolites in the positive and negative ion modes between children with GERD and HCs, which were mainly involved in arachidonic acid (AA), tyrosine, glutathione and caffeine metabolism.CONCLUSION: This study provides new evidence of the pathogenesis of GERD. There are significant differences in the gut microbiota, metabolites and metabolic pathways between HCs and children with GERD, and the differences in metabolites are related to specific changes in bacterial abundance. In the future, GERD may be treated by targeting specific bacteria related to AA metabolism.PMID:37900308 | PMC:PMC10613033 | DOI:10.3389/fcimb.2023.1267192

Characteristics of amino acid metabolism in colorectal cancer

Mon, 30/10/2023 - 11:00
World J Clin Cases. 2023 Sep 26;11(27):6318-6326. doi: 10.12998/wjcc.v11.i27.6318.ABSTRACTIn recent years, metabolomics research has become a hot spot in the screening and treatment of cancer. It is a popular technique for the quantitative characterization of small molecular compounds in biological cells, tissues, organs or organisms. Further study of the tumor revealed that amino acid changes may occur early in the tumor. The rapid growth and metabolism required for survival result in tumors exhibiting an increased demand for amino acids. An abundant supply of amino acids is important for cancer to maintain its proliferative driving force. Changes in amino acid metabolism can be used to screen malignant tumors and improve therapeutic outcomes. Therefore, it is particularly important to study the characteristics of amino acid metabolism in colorectal cancer. This article reviews several specific amino acid metabolism characteristics in colorectal cancer.PMID:37900242 | PMC:PMC10601002 | DOI:10.12998/wjcc.v11.i27.6318

Baseline metabolites could predict responders with hepatitis B virus-related liver fibrosis for entecavir or combined with FuzhengHuayu tablet

Mon, 30/10/2023 - 11:00
World J Hepatol. 2023 Sep 27;15(9):1043-1059. doi: 10.4254/wjh.v15.i9.1043.ABSTRACTBACKGROUND: After receiving entecavir or combined with FuzhengHuayu tablet (FZHY) treatment, some sufferers with hepatitis B virus (HBV)-related liver fibrosis could achieve a histological improvement while the others may fail to improve even worsen. Serum metabolomics at baseline in these patients who were effective in treatment remain unclear.AIM: To explore baseline serum metabolites characteristics in responders.METHODS: A total of 132 patients with HBV-related liver fibrosis and 18 volunteers as healthy controls were recruited. First, all subjects were divided into training set and validation set. Second, the included patients were subdivided into entecavir responders (E-R), entecavir no-responders (E-N), FZHY + entecavir responders (F-R), and FZHY + entecavir no-responders (F-N) following the pathological histological changes after 48 wk' treatments. Then, Serum samples of all subjects before treatment were tested by high performance liquid chromatography-tandem mass spectrometry (LC-MS) high-performance LC-MS. Data processing was conducted using multivariate principal component analysis and orthogonal partial least squares discriminant analysis. Diagnostic tests of selected differential metabolites were used for Boruta analyses and logistic regression.RESULTS: As for the intersection about differential metabolic pathways between the groups E-R vs E-N and F-R vs F-N, results showed that 4 pathways including linoleic acid metabolism, aminoacyl-tRNA biosynthesis, cyanoamino acid metabolism, alanine, aspartate and glutamate metabolism were screened out. As for the differential metabolites, these 7 intersected metabolites including hydroxypropionic acid, tyrosine, citric acid, taurochenodeoxycholic acid, benzoic acid, 2-Furoic acid, and propionic acid were selected.CONCLUSION: Our findings showed that 4 metabolic pathways and 7 differential metabolites had potential usefulness in clinical prediction of the response of entecavir or combined with FZHY on HBV fibrotic liver.PMID:37900214 | PMC:PMC10600694 | DOI:10.4254/wjh.v15.i9.1043

Significant dysregulation of lipid metabolism in patients with papillary thyroid carcinoma after thyroidectomy

Mon, 30/10/2023 - 11:00
Front Endocrinol (Lausanne). 2023 Oct 12;14:1223527. doi: 10.3389/fendo.2023.1223527. eCollection 2023.ABSTRACTINTRODUCTION: Thyroidectomy and thyrotropin suppressive therapy is the widely used surgical treatment for papillary thyroid carcinoma (PTC) patients. However, systematic metabolic changes of post-operative PTC patients were rarely reported.METHODS: Here, untargeted metabolomic detection of cohorts from PTC before (t0) and 1-month-after (t1) thyroidectomy, were performed to characterize circulating metabolic signatures after surgical treatment.RESULTS: Our results showed PTC patients exhibited lower thyroid stimulating hormone degree, higher total thyroxine, and significant lipid-related metabolic alternations after thyroidectomy, which included 97 upregulations (including 93 lipids) and 5 downregulations (including 2 lipids and 3 nucleotides). Enrichment of metabolic pathways mainly included biosynthesis of fatty acids, purine metabolism, and linoleic acid metabolism. We also demonstrated that differential surgical approaches (hemi- and total thyroidectomy) and post-operative complication phenotypes (insomnia, fatigue), might lead to characteristic metabolic signatures.DISCUSSION: This study revealed dynamic changes of metabolite characteristics of PTC patients after surgical treatment, which were associated with clinical thyroid function parameters, surgical approaches, and complication occurrence. It enlightened us to pay more attention on the post-operative metabolic dysregulation of PTC patients and their long-term qualities of life, so as to provide cautious clinical decisions on surgical choices, treatments, and follow-up details.PMID:37900134 | PMC:PMC10602777 | DOI:10.3389/fendo.2023.1223527

Association of sex hormones and sex hormone-binding globulin with liver fat in men and women: an observational and Mendelian randomization study

Mon, 30/10/2023 - 11:00
Front Endocrinol (Lausanne). 2023 Oct 13;14:1223162. doi: 10.3389/fendo.2023.1223162. eCollection 2023.ABSTRACTBACKGROUND: Sex hormones and sex hormone-binding globulin (SHBG) may play a role in fatty liver development. We sought to examine the association of various endogenous sex hormones, including testosterone (T), and SHBG with liver fat using complementary observational and Mendelian randomization (MR) analyses.METHODS: The observational analysis included a total of 2,239 participants (mean age 60 years; 35% postmenopausal women) from the population-based KORA study (average follow-up time: 6.5 years). We conducted linear regression analysis to investigate the sex-specific associations of sex hormones and SHBG with liver fat, estimated by fatty liver index (FLI). For MR analyses, we selected genetic variants associated with sex hormones and SHBG and extracted their associations with magnetic resonance imaging measured liver fat from the largest up to date European genome-wide associations studies.RESULTS: In the observational analysis, T, dihydrotestosterone (DHT), progesterone and 17α-hydroxyprogesterone (17-OHP) were inversely associated with FLI in men, with beta estimates ranging from -4.23 to -2.30 [p-value <0.001 to 0.003]. Whereas in women, a positive association of free T with FLI (β = 4.17, 95%CI: 1.35, 6.98) was observed. SHBG was inversely associated with FLI across sexes [men: -3.45 (-5.13, -1.78); women: -9.23 (-12.19, -6.28)]. No causal association was found between genetically determined sex hormones and liver fat, but higher genetically determined SHBG was associated with lower liver fat in women (β = -0.36, 95% CI: -0.61, -0.12).CONCLUSION: Our results provide suggestive evidence for a causal association between SHBG and liver fat in women, implicating the protective role of SHBG against liver fat accumulation.PMID:37900132 | PMC:PMC10611498 | DOI:10.3389/fendo.2023.1223162

Investigating the relationship between non-occupational pesticide exposure and metabolomic biomarkers

Mon, 30/10/2023 - 11:00
Front Public Health. 2023 Oct 11;11:1248609. doi: 10.3389/fpubh.2023.1248609. eCollection 2023.ABSTRACTThe relationship between pesticide exposures and metabolomics biomarkers is not well understood. We examined the changes in the serum metabolome (early biomarkers) and the metabolic pathways associated with various pesticide exposure scenarios (OPE: overall exposure, PEM: exposure in months, PEY: exposure in years, and PEU: reported specific pesticides use) using data from the Northern Finland Birth Cohort 1966 31-year cross-sectional examination. We utilized questionnaire data on pesticide exposures and serum samples for nuclear magnetic resonance (NMR)-based metabolomics analyses. For exposures and metabolites associations, participants size varied between 2,361 and 5,035. To investigate associations between metabolomics biomarkers and exposure to pesticide scenarios compared to those who reported no exposures multivariable regression analyses stratified by sex and adjustment with covariates (season of pesticide use, socioeconomic position (SEP), alcohol consumption, BMI, and latitude of residence) were performed. Multiple testing by Benjamini-Hochberg false discovery rate (FDR) correction applied. Pesticide exposures differed by sex, season of pesticide use, alcohol, SEP, latitude of residence. Our results showed that all pesticide exposure scenarios were negatively associated with decreased HDL concentrations across all lipoprotein subclasses in women. OPE, PEY, and PEU were associated with decreased branched-chain amino acid concentrations in men and decreased albumin concentrations in women. OPE, PEY and PEU were also associated with changes in glycolysis metabolites and ketone bodies in both sexes. Specific pesticides exposure was negatively associated with sphingolipids and inflammatory biomarkers in men. In women, OPE, PEM, and PEU were associated with decreased apolipoprotein A1 and increased apolipoprotein B/apolipoprotein A1 ratio. Our findings suggest that identification of early biomarkers of disease risk related to pesticide exposures can inform strategies to reduce exposure and investigate causal pathways. Women may be more susceptible to non-occupational pesticide exposures when compared to men, and future sex-specific studies are warranted.PMID:37900012 | PMC:PMC10602903 | DOI:10.3389/fpubh.2023.1248609

Widely targeted metabolomics reveals the effect of different raw materials and drying methods on the quality of instant tea

Mon, 30/10/2023 - 11:00
Front Nutr. 2023 Oct 11;10:1236216. doi: 10.3389/fnut.2023.1236216. eCollection 2023.ABSTRACTINTRODUCTION: Instant teas are particularly rich in tea polyphenols and caffeine and have great potential as food ingredients or additives to improve the quality of food and enhance their nutritional and commercial value.METHODS: To determine the relationships between raw material, drying method, and sensory and other quality attributes, instant teas were prepared from three tea varieties, namely black, green and jasmine tea, using two drying methods, namely spray-drying (SD) and freeze-drying (FD).RESULTS: Both the raw tea material and drying method influenced the quality of the finished instant teas. Black tea was quality stable under two drying, while green tea taste deteriorated much after SD. Jasmine tea must be produced from FD due to huge aroma deterioration after SD. FD produced instant tea with higher sensory quality, which was attributed to the lower processing temperature. Chemical compositional analysis and widely targeted metabolomics revealed that SD caused greater degradation of tea biochemical components. The flavonoids content changed markedly after drying, and metabolomics, combined with OPLS-DA, was able to differentiate the three varieties of tea. Instant tea preparations via SD often lost a large proportion of the original tea aroma compounds, but FD minimized the loss of floral and fruity aroma compounds. Changes in the tea flavonoids composition, especially during drying, contributed to the flavor development of instant tea.DISCUSSION: These results will provide an practicle method for high-quality instant tea production through choosing proper raw tea material and lowering down drying temperature with non-thermal technologies like FD.PMID:37899836 | PMC:PMC10600452 | DOI:10.3389/fnut.2023.1236216

Editorial: Metabolic response: nexus or nemesis for the understanding of sports nutrition and doping

Mon, 30/10/2023 - 11:00
Front Nutr. 2023 Oct 12;10:1306052. doi: 10.3389/fnut.2023.1306052. eCollection 2023.NO ABSTRACTPMID:37899835 | PMC:PMC10602773 | DOI:10.3389/fnut.2023.1306052

Soil cadmium pollution facilitated the invasion of alligator weed through enhanced herbivore resistance and competitive ability over a congeneric species

Mon, 30/10/2023 - 11:00
Plant Cell Environ. 2023 Oct 30. doi: 10.1111/pce.14747. Online ahead of print.ABSTRACTA number of invasive plant species, such as Alternanthera philoxeroides, have been documented to be able to accumulate trace metal elements in their tissues. Since metal accumulation in plants can serve as a defence against herbivores, we hypothesized that metal pollution will increase herbivore resistance of metal-accumulating invasive plant species and such a benefit will grant them a competitive advantage over local co-occurring plants. In this study, we compared the differences in plant growth and herbivore feeding preference between A. philoxeroides and its native congener Alternanthera sessilis in single and mixed cultures with and without soil cadmium (Cd) pollution. The results showed that A. philoxeroides plants were more tolerant to Cd stress and accumulated more Cd in the leaves than A. sessilis. Cd exposure increased the resistance of A. philoxeroides against a specialist and a generalist herbivore compared with A. sessilis. Competition experiments indicated that Cd stress largely increased the competitive advantage of A. philoxeroides over A. sessilis with or without herbivore pressures. The differences in herbivore resistance between the two plant species under soil Cd stress are most likely due to the deterring effect of Cd accumulation and Cd-enhanced mechanical defences rather than changes in leaf specialized metabolites.PMID:37899642 | DOI:10.1111/pce.14747

Lipidome and metabolome analyses reveal metabolic alterations associated with MCF-7 apoptosis upon 4-hydroxytamoxifen treatment

Mon, 30/10/2023 - 11:00
Sci Rep. 2023 Oct 29;13(1):18549. doi: 10.1038/s41598-023-45764-2.ABSTRACT4-hydroxytamoxifen (OHT) is an anti-cancer drug that induces apoptosis in breast cancer cells. Although changes in lipid levels and mitochondrial respiration have been observed in OHT-treated cells, the overall mechanisms underlying these metabolic alterations are poorly understood. In this study, time-series metabolomics and lipidomics were used to analyze the changes in metabolic profiles induced by OHT treatment in the MCF-7 human breast cancer cell line. Lipidomic and metabolomic analyses revealed increases in ceramide, diacylglycerol and triacylglycerol, and decreases in citrate, respectively. Gene expression analyses revealed increased expression of ATP-dependent citrate lyase (ACLY) and subsequent fatty acid biosynthetic enzymes, suggesting that OHT-treated MCF-7 cells activate citrate-to-lipid metabolism. The significance of the observed metabolic changes was evaluated by co-treating MCF-7 cells with OHT and ACLY or a diacylglycerol O-acyltransferase 1 (DGAT1) inhibitor. Co-treatment ameliorated cell death and reduced mitochondrial membrane potential compared to that in OHT treatment alone. The inhibition of cell death by co-treatment with an ACLY inhibitor has been observed in other breast cancer cell lines. These results suggest that citrate-to-lipid metabolism is critical for OHT-induced cell death in breast cancer cell lines.PMID:37899460 | DOI:10.1038/s41598-023-45764-2

Neuronal progenitor cells-based metabolomics study reveals dysregulated lipid metabolism and identifies putative biomarkers for CLN6 disease

Mon, 30/10/2023 - 11:00
Sci Rep. 2023 Oct 29;13(1):18550. doi: 10.1038/s41598-023-45789-7.ABSTRACTNeuronal ceroid lipofuscinosis 6 (CLN6) is a rare and fatal autosomal recessive disease primarily affecting the nervous system in children. It is caused by a pathogenic mutation in the CLN6 gene for which no therapy is available. Employing an untargeted metabolomics approach, we analyzed the metabolic changes in CLN6 subjects to see if this system could potentially yield biomarkers for diagnosis and monitoring disease progression. Neuronal-like cells were derived from human fibroblast lines from CLN6-affected subjects (n = 3) and controls (wild type, n = 3). These were used to assess the potential of a neuronal-like cell-based metabolomics approach to identify CLN6 distinctive and specific biomarkers. The most impacted metabolic profile is associated with sphingolipids, glycerophospholipids metabolism, and calcium signaling. Over 2700 spectral features were screened, and fifteen metabolites were identified that differed significantly between both groups, including the sphingolipids C16 GlcCer, C24 GlcCer, C24:1 GlcCer and glycerophospholipids PG 40:6 and PG 40:7. Of note, these fifteen metabolites were downregulated in the CLN6 disease group. This study is the first to analyze the metabolome of neuronal-like cells with a pathogenic mutation in the CLN6 gene and to provide insights into their metabolomic alterations. This could allow for the development of novel biomarkers for monitoring CLN6 disease.PMID:37899458 | DOI:10.1038/s41598-023-45789-7

Quantitative analysis of the glutathione pathway cellular metabolites by targeted liquid chromatography - tandem mass spectrometry

Sun, 29/10/2023 - 11:00
J Sep Sci. 2023 Oct 29:e2300780. doi: 10.1002/jssc.202300780. Online ahead of print.ABSTRACTGlutathione, its biosynthesis intermediates and other thiol metabolites are of central relevance for the redox homeostasis of cells. Their analysis is critical due to the facile interconversion of redox pairs during sampling, sample preparation, and data acquisition, in particular in the electrospray ionization interface. In this work we propose a fast targeted LC-MS/MS method to accurately analyze 14 metabolites from the glutathione pathway. N-Ethylmaleimide reagent is added with the extraction solvent and instantly stabilizes the thiol-redox state by derivatization. Liquid chromatographic separation of the analytes was performed on a sub-2μm superficially porous HILIC column with sulfobetaine chemistry. Tandem MS with triple-quadrupole mass spectrometry in multiple-reaction monitoring acquisition mode allowed sensitive detection of the targeted metabolites with LOQs in the range of 5-25 nM. Run times of 3 min enable a high throughput analysis of cellular samples. For calibration a 13 C-labelled cell extract was used as internal standard. The method was validated and the concentrations of glutathione and its biosynthesis intermediates determined in HeLa cells. This article is protected by copyright. All rights reserved.PMID:37898873 | DOI:10.1002/jssc.202300780

Systems biology of industrial oxytetracycline production in Streptomyces rimosus: the secrets of a mutagenized hyperproducer

Sun, 29/10/2023 - 11:00
Microb Cell Fact. 2023 Oct 28;22(1):222. doi: 10.1186/s12934-023-02215-x.ABSTRACTBACKGROUND: Oxytetracycline which is derived from Streptomyces rimosus, inhibits a wide range of bacteria and is industrially important. The underlying biosynthetic processes are complex and hinder rational engineering, so industrial manufacturing currently relies on classical mutants for production. While the biochemistry underlying oxytetracycline synthesis is known to involve polyketide synthase, hyperproducing strains of S. rimosus have not been extensively studied, limiting our knowledge on fundamental mechanisms that drive production.RESULTS: In this study, a multiomics analysis of S. rimosus is performed and wild-type and hyperproducing strains are compared. Insights into the metabolic and regulatory networks driving oxytetracycline formation were obtained. The overproducer exhibited increased acetyl-CoA and malonyl CoA supply, upregulated oxytetracycline biosynthesis, reduced competing byproduct formation, and streamlined morphology. These features were used to synthesize bhimamycin, an antibiotic, and a novel microbial chassis strain was created. A cluster deletion derivative showed enhanced bhimamycin production.CONCLUSIONS: This study suggests that the precursor supply should be globally increased to further increase the expression of the oxytetracycline cluster while maintaining the natural cluster sequence. The mutagenized hyperproducer S. rimosus HP126 exhibited numerous mutations, including large genomic rearrangements, due to natural genetic instability, and single nucleotide changes. More complex mutations were found than those typically observed in mutagenized bacteria, impacting gene expression, and complicating rational engineering. Overall, the approach revealed key traits influencing oxytetracycline production in S. rimosus, suggesting that similar studies for other antibiotics could uncover general mechanisms to improve production.PMID:37898787 | DOI:10.1186/s12934-023-02215-x

Metabolomic and Lipidomic Signature of Skeletal Muscle with Constitutively Active mTORC1

Sat, 28/10/2023 - 12:00
J Nutr. 2023 Oct 26:S0022-3166(23)72670-9. doi: 10.1016/j.tjnut.2023.10.016. Online ahead of print.ABSTRACTBACKGROUND: Regulation of mTORC1 plays an important role in aging and nutrition. For example, caloric restriction reduces mTORC1 signaling and extends lifespan, whereas nutrient abundance and obesity increase mTORC1 signaling and reduce lifespan. Skeletal muscle specific knockout of DEP domain-containing 5 protein (DEPDC5) results in constitutively active mTORC1 signaling, muscle hypertrophy and an increase in mitochondrial respiratory capacity. The metabolic profile of skeletal muscle, in the setting of hyperactive mTORC1 signaling, is not well known.OBJECTIVES: To determine the metabolomic and lipidomic signature in skeletal muscle from female and male wildtype and DEPDC5 knockout mice.METHODS: Tibialis anterior (TA) muscles from wildtype (WT) and transgenic (conditional skeletal muscle specific DEPDC5 knockout, KO) were obtained from female (F) and male (M) adult mice. Polar metabolites and lipids were extracted using a Bligh-Dyer extraction from 5 samples per group and identified and quantified by LC-MS/MS. Resulting analyte peak areas were analyzed with t-test, ANOVA, and volcano plots for group comparisons (e.g., WT vs KO) and multivariate statistical analysis for genotype and sex comparisons.RESULTS: A total of 162 polar metabolites (organic acids, amino acids and amines and acyl carnitines) and 1141 lipid metabolites were detected in TA samples by LC-MS/MS. Few polar metabolites showed significant differences in KO muscles compared to WT within the same sex group. P-aminobenzoic acid, β-alanine and dopamine were significantly higher in KO male muscle whereas erythrose- 4-phosphate and oxoglutaric acid were significantly reduced in KO females. The lipidomic profile of the KO groups revealed an increase of muscle phospholipids and reduced triacylglycerol and diacylglycerol compared to the WT groups.CONCLUSIONS: Sex differences were detected in polar metabolome and lipids were dependent on genotype. Metabolomic profile of mice with hyperactive skeletal muscle mTORC1 is consistent with an upregulation of mitochondrial function and amino acid utilization for protein synthesis.PMID:37898335 | DOI:10.1016/j.tjnut.2023.10.016

Insights into the mechanism of action of the chlorophyll derivative 13-<sup>2</sup>-hydroxypheophytine a on reducing neutral lipid reserves in zebrafish larvae and mice adipocytes

Sat, 28/10/2023 - 12:00
Eur J Pharmacol. 2023 Oct 26:176158. doi: 10.1016/j.ejphar.2023.176158. Online ahead of print.ABSTRACTObesity is a worldwide epidemic and natural products may hold promise in its treatment. The chlorophyll derivative 13-2-hydroxypheophytine (hpa) was isolated in a screen with zebrafish larvae to identify lipid reducing molecules from cyanobacteria. However, the mechanisms underlying the lipid-reducing effects of hpa in zebrafish larvae remain poorly understood. Thus, investigating the mechanism of action of hpa and validation in other model organisms such as mice represents important initial steps. In this study, we identified 14 protein targets of hpa in zebrafish larvae by thermal proteome profiling, and selected two targets (malate dehydrogenase and pyruvate kinase) involved in cellular metabolism for further validation by enzymatic measurements. Our findings revealed a dose-dependent inhibition of pyruvate kinase by hpa exposure using protein extracts of zebrafish larvae in vitro, and in exposure experiments from 3 to 5 days post fertilization in vivo. Analysis of untargeted metabolomics of zebrafish larvae detected 940 mass peaks (66 increased, 129 decreased) and revealed that hpa induced the formation of various phospholipid species (phosphoinositol, phosphoethanolamine, phosphatidic acid). Inter-species validation showed that brown adipocytes exposed to hpa significantly reduced the size of lipid droplets, increased maximal mitochondrial respiratory capacity, and the expression of PPARy during adipocyte differentiation. In line with our data, previous work described that reduced pyruvate kinase activity lowered hepatic lipid content via reduced pyruvate and citrate, and improved mitochondrial function via phospholipids. Thus, our data provide new insights into the molecular mechanism underlying the lipid reducing activities of hpa in zebrafish larvae, and species overlapping functions in reduction of lipids.PMID:37898286 | DOI:10.1016/j.ejphar.2023.176158

Determination of well-being-related markers in nails by liquid chromatography tandem mass spectrometry

Sat, 28/10/2023 - 12:00
Ecotoxicol Environ Saf. 2023 Oct 26;267:115586. doi: 10.1016/j.ecoenv.2023.115586. Online ahead of print.ABSTRACTWell-being is a multifactorial positive state that is highly influenced by some endogenous molecules that control happiness and euphoric feelings. These molecules, e.g., neurotransmitters, hormones and their derivatives, play a crucial role in metabolism and may be referred to as "well-being-related markers". The deregulation of well-being-related markers can lead to organism malfunctions and life-threatening states. In this research, we aimed to evaluate the potential of nails for the chronic production of several well-being-related markers. For this purpose, we developed an LCMS /MS-based method for the determination of 10 well-being-related markers, including melatonin, serotonin, cortisol, kynurenine and several precursors and metabolites. The method was optimized regarding different analytical steps: required sample amount, extraction time, number of required extractions, preconcentration, injection volume and MS conditions. Method validation was performed by two different approaches: (i) using surrogate nail matrix and (ii) using authentic nail samples by standard additions. The method was found to be linear in the expected endogenous range and sensitive enough to determine the low endogenous concentration levels in nails. Accuracy and precision were appropriate in both validation approaches. As proof of concept, the method was used (i) to correlate fingernail and toenail levels for all metabolites in 22 volunteers, (ii) to establish the endogenous concentration range of all metabolites in females (n = 50) and males (n = 34) and (iii) to correlate the metabolite levels with age. For some metabolites, the calculated ranges have been reported for the first time. In summary, the present strategy to evaluate well-being-related markers in nails may be a useful tool for the evaluation of the production of these important compounds with high potential for a wide range of clinical purposes.PMID:37897979 | DOI:10.1016/j.ecoenv.2023.115586

Environmental concentrations of cadmium and zinc and associating metabolomics profile alternations in urine of pregnant women in the first trimester: A prospective cohort study in Taiyuan, North China

Sat, 28/10/2023 - 12:00
Ecotoxicol Environ Saf. 2023 Oct 26;267:115611. doi: 10.1016/j.ecoenv.2023.115611. Online ahead of print.ABSTRACTespecially to pregnant women. In recent years, zinc (Zn) supplementation has attracted increasing attention among pregnant women. Thus, understanding the effects and interactions of Cd and Zn in pregnant women is critical. This study aimed to assess the urinary levels of Cd and Zn in pregnant women during early pregnancy, examine associated alterations in urine metabolomics, and identify potential metabolic biomarkers among distinct Cd and Zn groups. Urine samples from 185 pregnant women were collected, and inductively coupled plasma mass spectrometry (ICP-MS) was used to detect Cd and Zn contents. The women were then divided into four groups according to median contents of Cd and Zn. Alterations in the metabolite profile were assessed using a liquid chromatograph mass spectrometer (LC-MS). The results showed that the gravidity of pregnant women was closely related to urinary Cd levels and that the urinary Zn contents of pregnant women with morning sickness in the first trimester were lower than that of non-morning-sick pregnant women. A total of 51 metabolites exhibited significant differential expression in the high level of Cd and Zn (HCdHZn) compared with low level of Cd and Zn (LCdLZn), the diagnostic performance of these 51 metabolites were assessed using receiver operating characteristic curve analysis and revealed that octadecylamine was a promising diagnostic indicator for evaluating the combined effects of Zn and Cd. Metabolomics analysis showed that the arginine and proline pathways were upregulated in HCdHZn compared with that in LCdLZn, suggesting a potential risk of obesity. Although higer levels of bovinic acid in HCdHZn vs. HCdLZn (high level of Cd and low level of Zn) indicated that Zn has antioxidant and anti-inflammatory properties, excessive Zn may still cause harmful effect to the human health and should be supplemented with caution. The study findings may be valuable for potential risk ahissessment of the combined effects of Cd-Zn and their interactions in pregnant women.PMID:37897977 | DOI:10.1016/j.ecoenv.2023.115611

Protein-coding potential of non-canonical open reading frames in human transcriptome

Sat, 28/10/2023 - 12:00
Biochem Biophys Res Commun. 2023 Oct 13;684:149040. doi: 10.1016/j.bbrc.2023.09.068. Online ahead of print.ABSTRACTIn recent years, proteogenomics and ribosome profiling studies have identified a large number of proteins encoded by noncoding regions in the human genome. They are encoded by small open reading frames (sORFs) in the untranslated regions (UTRs) of mRNAs and long non-coding RNAs (lncRNAs). These sORF encoded proteins (SEPs) are often <150AA and show poor evolutionary conservation. A subset of them have been functionally characterized and shown to play an important role in fundamental biological processes including cardiac and muscle function, DNA repair, embryonic development and various human diseases. How many novel protein-coding regions exist in the human genome and what fraction of them are functionally important remains a mystery. In this review, we discuss current progress in unraveling SEPs, approaches used for their identification, their limitations and reliability of these identifications. We also discuss functionally characterized SEPs and their involvement in various biological processes and diseases. Lastly, we provide insights into their distinctive features compared to canonical proteins and challenges associated with annotating these in protein reference databases.PMID:37897910 | DOI:10.1016/j.bbrc.2023.09.068

Effect of postharvest processing on quality traits of Radix Gentianae Macrophyllae: A integrative analysis of metabolomics and proteomics

Sat, 28/10/2023 - 12:00
Plant Physiol Biochem. 2023 Oct 15;204:108099. doi: 10.1016/j.plaphy.2023.108099. Online ahead of print.ABSTRACTThe dried roots and rhizomes of Radix Gentianae Macrophyllae are widely used as food material or medicinal crops. "Sweating" is a traditional postharvest processing method, the basic processing procedure consists of softening, stacking and drying. The aim of this paper is to unveil the scientific connotation responsible for the "Sweating" processing in Radix Gentianae Macrophyllae during postharvest. Thus, the effect of different postharvest processing methods on the metabolic pathways of Radix Gentiasnae Macrophyllae was studied by the non-targeted metabolomic technique in combination with the label-free proteomics approach. The results showed that the differentially accumulated metabolites and abundant proteins were mainly enriched in the pathways of phenylalanine, tyrosine and tryptophan biosynthesis, polyphenols and terpenoids biosynthesis. "Sweating" has a greater up-regulation effect on these pathways than "Non-sweating", and can induce protein expression and metabolite accumulation associated with the quality traits of Radix Gentianae Macrophyllae. The results provide a detailed explanation of the scientific connotation of crucial steps of "Sweating" processing wherein opportunities existed for taking appropriate measures to enhance the accumulation of bioactive ingredients. These findings will serve as significant references for enhancing the postharvest processing technology of Radix Gentianae Macrophyllae and similar plants, resulting in higher product quality for food or plant materials production.PMID:37897890 | DOI:10.1016/j.plaphy.2023.108099

Investigation of the effect of over-fired drying on the taste and aroma of Lu'an Guapian tea using metabolomics and sensory histology techniques

Sat, 28/10/2023 - 12:00
Food Chem. 2023 Oct 25;437(Pt 1):137851. doi: 10.1016/j.foodchem.2023.137851. Online ahead of print.ABSTRACTLu'an Guapian (LAGP) tea, a representative Chinese roasted green tea, undergoes significant changes in taste and aroma during over-fired drying. However, limited studies have been conducted on these effects. This study employed metabolomics and sensory histology techniques to analyze non-volatile and volatile compounds the second drying and pulley liquefied gas drying (PLD) samples. The results revealed that after PLD, the samples exhibited lower umami, bitterness, and astringency; whereas floral, sweet, roasted, cooked corn-like, and cooked chestnut-like aromas became stronger. Among them, the content of (-)-epigallocatechin gallate, glutamic acid, and theogallin, which were closely related to taste, decreased by 4.5 %, 12.3 %, and 10.4 %, respectively. Eight key aroma components were identified as the main contributors to the sample aroma changes: (E)-β-ionone, dimethyl sulfide, (E,E)-2,4-heptadienal, geraniol, linalool, benzeneacetaldehyde, 2-ethyl-3,5-dimethylpyrazine, and hexanal. This study provides a theoretical basis for enhancing the quality of LAGP teas.PMID:37897824 | DOI:10.1016/j.foodchem.2023.137851

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