Targeted Metabolite Profiling Research Articles

LC-MS-Based Targeted Metabolite Profiling and Atomic Absorption Spectrophotmeter Based Mineral Analysis of Methanol Extract of Terminalia bellerica (Gaertn.) Leaves

Aim: Natural products in medicinal plants are one of the important sources for modern drug discovery. Plant-based formulations play an important role in management of numerous diseases. To discover the medical potential of natural products, it is important to understand the bioactive compounds present in medicinal plants. Hence the aim of the study is to identify the volatile compounds by LC –MS analysis and minerals by AAS assay present in the leaves of Terminalia bellerica. Study Design: This study investigates metabolites present in leaves of Terminalia bellerica have been studied by AAS and LC-MS techniques. Results: The AAS assay showed that the leaves of T. bellerica are particularly rich in iron (260 mg/kg) compared to their zinc, manganese, and copper content. Meanwhile, the LC-MS analysis identified chebulinic acid, chebulagic acid, punicafolin, myricetin, rutin, and galloyl punicalagin as being present in the highest concentrations. Conclusion: Results highlighted presence of considerable amount of iron and other minerals in methanolic extract of the leaves of T. bellerica, it may be used in the development of health supplements. The results also revealed that bioactive compounds in the methanolic extracts of T. bellerica leave used for drug design and making it safe to human health.

Limited Effectiveness of Penicillium camemberti in Preventing the Invasion of Contaminating Molds in Camembert Cheese.

Mold-ripened cheese acquires a distinctive aroma and texture from mold cultures that mature on a fresh cheese wheel. Owing to its high moisture content (aw = 0.95) and pliability, soft cheese is prone to contamination. Many contaminating mold species are unable to grow at colder temperatures, and the lactic acid produced by the cheese bacteria inhibits further infiltration. Thus, Camembert cheese is generally well protected against contamination by a wide range of species. In this study, cocultures of Penicillium camemberti and widely distributed mycotoxin-producing mold species were incubated on different types of agars, and purchased Camembert samples were deliberately contaminated with mycotoxin-producing mold species capable of growing at both 25 °C and 4 °C. The production of mycotoxins was then monitored by the extraction of the metabolites and their subsequent measurement by means of a liquid chromatography-tandem mass spectrometry (LC-MS/MS) based targeted metabolite profiling approach. The production of cyclopiazonic acid (CPA) was highly dependent on the species cocultivated with Penicillium camemberti, the temperature and the substrate. Contamination of Camembert cheese with Penicillium chrysogenum, Mucor hiemalis, or Penicillium glabrum induced CPA production at 25 °C. Although mold growth on cheese was not always evident on biofilms for certain cultures, except for Penicillium citrinum, which stained the monosaccharide agar yellow, mycotoxins were detected in many agar and cheese samples, as in all monosaccharide agar samples. In conclusion, cheese should be immediately discarded upon the first appearance of mold.

Insights into the role of GhCYP and GhTPS in the gossypol biosynthesis pathway via a multiomics and functional-based approach in cotton

Two Cotton Research Institute (CRI) near-isogenic lines, CRI-12 glanded and CRI-12 glandless, were used to pinpoint potential genes and metabolic pathways linked to gossypol biosynthesis through transcriptome sequencing. We discovered more than 235 million clean reads and 1,184 differentially expressed genes (DEGs). Consecutively, we conducted a weighted gene co-expression network analysis and found strong correlation of white and yellow modules containing GhTPS (GH_D09G0090) and GhCYP (GH_D05G2016) hub genes with the gossypol content. The importance of the GhTPS and GhCYP genes was demonstrated using RT-qPCR, virus-induced gene silencing (VIGS) and target metabolite analysis. Silencing of these genes resulted in fewer glands on both leaves and stems two weeks after the infection, as compared to the wild type. In addition, a total of 152 metabolites were identified through targeted metabolite profiling. Differential metabolite screening revealed 12 and 18 significantly different metabolites in TRV:GhTPS and TRV:GhCYP plants vs. control group, respectively, showing reduction in accumulation of metabolites compared to the control. The content of hemigossypol, the final product of gossypol biosynthesis, was also reduced, as revealed by target metabolite analysis, suggesting the role of these genes in the gossypol biosynthetic pathway. Furthermore, a highly significant difference in gossypol content between the glanded and glandless lines was recorded. The findings of this study reveal a strong link between the gossypol content and GhTPS and GhCYP hub genes, suggesting their role in the gossypol biosynthetic pathway to reduce the accumulation of hemigossypol, which may offer new comprehension into the regulatory checkpoints of the gossypol biosynthesis pathway in cotton.

Comparative Metabolic Profiling of Different Colored Rice Grains Reveals the Distribution of Major Active Compounds and Key Secondary Metabolites in Green Rice.

Pigmented rice grains are important resources for health and nutritional perspectives. Thus, a thorough dissection of the variation of nutrients and bioactive metabolites in different colored rice is of global interest. This study applied LC-MS-based widely targeted metabolite profiling and unraveled the variability of metabolites and nutraceuticals in long grain/non-glutinous black (BR), red (RR), green (GR), and white rice (WR) grains. We identified and classified 1292 metabolites, including five flavonoid compounds specific to BR. The metabolite profiles of the four rice grains showed significant variation, with 275-543 differentially accumulated metabolites identified. Flavonoid (flavone, flavonol, and anthocyanin) and cofactor biosynthesis were the most differentially regulated pathways among the four rice types. Most bioactive flavonoids, anthocyanidins (glycosylated cyanidins and peonidins), phenolic acids, and lignans had the highest relative content in BR, followed by RR. Most alkaloids, amino acids and derivatives, lipids, and vitamins (B6, B3, B1, nicotinamide, and isonicotinic acid) had higher relative contents in GR than others. Procyanidins (B1, B2, and B3) had the highest relative content in RR. In addition, we identified 25 potential discriminatory biomarkers, including fagomine, which could be used to authenticate GR. Our results show that BR and RR are important materials for medicinal use, while GR is an excellent source of nutrients (amino acids and vitamins) and bioactive alkaloids. Moreover, they provide data resources for the science-based use of different colored rice varieties in diverse industries.

The road less taken: Dihydroflavonol 4-reductase inactivation and delphinidin anthocyanin loss underpins a natural intraspecific flower colour variation.

Visual cues are of critical importance for the attraction of animal pollinators, however, little is known about the molecular mechanisms underpinning intraspecific floral colour variation. Here, we combined comparative spectral analysis, targeted metabolite profiling, multi-tissue transcriptomics, differential gene expression, sequence analysis and functional analysis to investigate a bee-pollinated orchid species, Glossodia major with common purple- and infrequent white-flowered morphs. We found uncommon and previously unreported delphinidin-based anthocyanins responsible for the conspicuous and pollinator-perceivable colour of the purple morph and three genetic changes underpinning the loss of colour in the white morph - (1) a loss-of-function (LOF; frameshift) mutation affecting dihydroflavonol 4-reductase (DFR1) coding sequence due to a unique 4-bp insertion, (2) specific downregulation of functional DFR1 expression and (3) the unexpected discovery of chimeric Gypsy transposable element (TE)-gene (DFR) transcripts with potential consequences to the genomic stability and post-transcriptional or epigenetic regulation of DFR. This is one of few known cases where regulatory changes and LOF mutation in an anthocyanin structural gene, rather than transcription factors, are important. Furthermore, if TEs prove to be a frequent source of mutation, the interplay between environmental stress-induced TE evolution and pollinator-mediated selection for adaptive colour variation may be an overlooked mechanism maintaining floral colour polymorphism in nature.

Metabolite Profiling, Biological and Molecular Analyses Validate the Nutraceutical Potential of Green Seaweed Acrosiphoniaorientalis for Human Health.

Seaweeds have proven to be nutrient-dense and are rich in antioxidants, like phenolics, flavonoids, and other essential metabolites that help to provide their medicinal benefits. Non-targeted metabolite profiling of the tropical green seaweed Acrosiphonia orientalis showed the presence of numerous groups of contents, including sugars, essential amino acids, and fatty acids. Targeted metabolite profiling using HPLC identified 17 amino acids. The extract exhibited a very low half-maximal effective concentration (EC50) dosage for HeLa and Huh-7 cell lines, indicating a high likelihood of anticancer properties. A significant positive correlation was found between biological activities, such as antioxidation, scavenging, and reducing power with the phenolic and flavonoid contents. The extract revealed augmentation of proliferation in selected cervical cells, as it upregulated p53 1.3-fold, and downregulated important cancerous genes such as Cas-3 and DNMT 12- and 8-fold, respectively. An approximate 55-fold downregulation was observed in selected hepatic cell lines. Microarray analysis of hepatic cells indicated 0.27% and 0.07% upregulation of coding and non-coding genes, respectively, and 0.41% and 0.13% downregulation of coding and non-coding genes, respectively. As a consequence, it can be said that A. orientalis has possible medicinal use, such as anticancer activity, and therefore may be an intriguing food component that has potential as a regular dietary supplement.

Targeted metabolite profiling of Salvia rosmarinus Italian local ecotypes and cultivars and inhibitory activity against Pectobacterium carotovorum subsp. carotovorum.

The development of agriculture in terms of sustainability and low environmental impact is, at present, a great challenge, mainly in underdeveloped and marginal geographical areas. The Salvia rosmarinus "Eretto Liguria" ecotype is widespread in Liguria (Northwest Italy), and farmers commonly use it by for cuttings and for marketing. In the present study, this ecotype was characterized in comparison with other cultivars from the same geographical region and Campania (Southern Italy), with a view to application and registration processes for the designation of protected geographical indications. Moreover, the possibility of using the resulting biomass after removing cuttings or fronds as a source of extracts and pure compounds to be used as phytosanitary products in organic farming was evaluated. Specifically, the potential of rosemary extracts and pure compounds to prevent soft rot damage was then tested. A targeted NMR metabolomic approach was employed, followed by multivariate analysis, to characterize the rosemary accessions. Bacterial soft rot assay and disk diffusion test were carried out to evaluate the activity of extracts and isolated compounds against Pectobacterium carotovorum subsp. carotovorum. Enzymatic assay was performed to measure the in vitro inhibition of the pectinase activity produced by the selected pathogen. Molecular docking simulations were used to explore the possible interaction of the selected compounds with the pectinase enzymes. The targeted metabolomic analysis highlighted those different geographical locations can influence the composition and abundance of bioactive metabolites in rosemary extracts. At the same time, genetic factors are important when a single geographical area is considered. Self-organizing maps (SOMs) showed that the accessions of "Eretto Liguria" appeared well characterized when compared to the others and had a good content in specialized metabolites, particularly carnosic acid. Soft rotting Enterobacteriaceae belonging to the Pectobacterium genus represent a serious problem in potato culture. Even though rosemary methanolic extracts showed a low antibacterial activity against a strain of Pectobacterium carotovorum subsp. carotovorum in the disk diffusion test, they showed ability in reducing the soft rot damage induced by the bacterium on potato tissue. 7-O-methylrosmanol, carnosol and isorosmanol appeared to be the most active components. In silico studies indicated that these abietane diterpenoids may interact with P. carotovorum subsp. carotovorum pectate lyase 1 and endo-polygalacturonase, thus highlighting these rosemary components as starting points for the development of agents able to prevent soft rot progression.

Kinetic simulation of convection drying for its effect on nutritional, functional, and phytochemical profiling of Colocasia antiquorum and Nasturtium officinale leaves of the Himalayan region

AbstractColocasia antiquorum (Taro) and Nasturtium officinale (Watercress) leaves are highly potent indigenous crops with rich nutritional and phytochemical profiles. The leaves were mathematically modeled, using thin‐layer models, to study the process kinetics for convection drying and its cumulative effect on the physico‐chemical, nutritional, functional, phytochemical, and targeted metabolite profiles. The best‐fit model for C. antiquorum was the two‐term model, whereas for N. officinale, Page model showed the best adequacy with the determination of least Chi‐square and high co‐efficient along with root mean square error. The color and pigment degradation were the prominent changes observed in the quality characteristics of C. antiquorum and N. officinale leaves depicted by the color degradation study. The nutritional composition was significantly affected by the convective drying with reduced ash, protein, and phenolic contents. The extraction yields were however higher along with some elevated metabolite contents estimated for convection drying in comparison to freeze‐dried leaves.Practical applicationsThis study explores the drying kinetic behavior for the progression of moisture loss occurring over the drying time and moderation of drying to achieve a desired moisture content at certain drying rates. The quality characterization before and after selected drying conditions revealed the sensitivity of the chlorophyll/carotenoid pigments, nutrients, and phenolics retained significantly after the end point of drying. Also, the effect of convective drying on the physicochemical, nutritional, and phytochemical contents was significant, which edifies the extent of alterations occurring due to convective drying on the quality characteristics of Colocasia antiquorum (Taro) and Nasturtium officinale (Watercress) leaves. Consequently, these two leafy vegetables hold good potential as crops to be promoted for nutritional security and health and wellness due to their rich bioactive profiles.

HAD2 Regulates Central Carbon Metabolism in Malaria Parasite P. falciparum

Abstract Background The malaria parasite Plasmodium falciparum causes nearly half a million deaths each year. Widespread resistance to antimalarials prompts an urgent need for novel drug targets both unique and essential to the parasite. The methylerythritol (MEP) pathway of isoprenoid biosynthesis fits this profile. Previous studies showed that P. falciparum controls substrates available to the MEP pathway via the phosphatase HAD2. A point mutation in the HAD2 gene was found to misdirect metabolites from glycolysis to the MEP pathway. Elucidating the mechanism by which HAD2 controls central carbon metabolism will inform our understanding of essential parasite biology and the potential for resistance to metabolic inhibitors under development. Methods In this work, we have generated a P. falciparum strain in which HAD2 expression is under control of anhydrous tetracycline (aTc), such that growth in the absence of aTc leads to loss of HAD2 expression. We have used this tunable knockdown strain to regulate expression of HAD2 in asexual-stage parasites to test whether HAD2 is required for glycolytic and isoprenoid biosynthesis homeostasis. Asexual parasite growth rate was measured through flow cytometry, and dose-response assays were performed against a competitive small molecule inhibitor of the MEP pathway. Targeted metabolite profiling with liquid chromatography-tandem mass spectrometry (LC-MS/MS) is in process to quantify glycolytic and isoprenoid metabolites in this knockdown strain. Results Knockdown parasites off aTc exhibited reduced growth rates relative to the wild-type strain. These parasites also exhibited a 4-fold increase in resistance to a competitive inhibitor of the MEP pathway compared to the wild-type [0.755 +/- 0.12 µM vs. 3.201 +/- 0.35 µM; p<0.001). Conclusions Reduced expression of HAD2 confers resistance to a competitive inhibitor of the MEP pathway, consistent with the effect of elevated levels of MEP pathway intermediates as a result of disrupted homeostasis. Ongoing work will use this strain to understand the role of HAD2 on parasite metabolism, parasite growth, and drug resistance.

The impact of a single dose of whey protein on glucose flux and metabolite profiles in normoglycemic males: insights into glucagon and insulin biology.

Protein ingestion concurrently stimulates euglycemic glucagon and insulin secretion, a response that is particularly robust with rapidly absorbing proteins. Previously, we have shown that ingestion of repeated doses of rapidly absorbing whey protein equally stimulated endogenous glucose production (EGP) and glucose disposal (Rd), thus explaining the preservation of euglycemia. Here, we aimed to determine if a smaller single dose of whey could elicit a large enough glucagon and insulin response to stimulate glucose flux. Therefore, in normoglycemic young adult males (N=10; Age ~26; BMI ~25), using [6,6-2H2] glucose tracing and quantitative targeted metabolite profiling, we determined the metabolic response to a single 25g 'standard' dose of whey protein. Whey protein ingestion did not alter glycemia, but increased circulating glucagon (peak 4-fold basal), insulin (peak 6-fold basal), amino acids and urea while also reducing free fatty acid and glycerol concentrations. Interestingly, the postprandial insulin response was driven by both a stimulation of insulin secretion and marked reduction in hepatic insulin clearance. Whey protein ingestion resulted in a modest stimulation of EGP and Rd, both peaking at ~20% above baseline 1 h after protein ingestion. These findings demonstrate that the ingestion of a single standard serving of whey protein can induce a euglycemic glucagon and insulin response that stimulates glucose flux. We speculate on a theory that could potentially explain how glucagon and insulin synergistically provide hardwired control of nitrogen and glucose homeostasis.

Flavonols affect the interrelated glucosinolate and camalexin biosynthetic pathways in Arabidopsis thaliana.

Flavonols are structurally and functionally diverse biomolecules involved in plant biotic and abiotic stress tolerance, pollen development, and inhibition of auxin transport. However, their effects on global gene expression and signaling pathways are unclear. To explore the roles of flavonol metabolites in signaling, we performed comparative transcriptome and targeted metabolite profiling of seedlings from the flavonol-deficient Arabidopsis loss-of-function mutant flavonol synthase1 (fls1) with and without exogenous supplementation of flavonol derivatives (kaempferol, quercetin, and rutin). RNA-seq results indicated that flavonols modulate various biological and metabolic pathways, with significant alterations in camalexin and aliphatic glucosinolate synthesis. Flavonols negatively regulated camalexin biosynthesis but appeared to promote the accumulation of aliphatic glucosinolates via transcription factor-mediated up-regulation of biosynthesis genes. Interestingly, upstream amino acid biosynthesis genes involved in methionine and tryptophan synthesis were altered under flavonol deficiency and exogenous supplementation. Quercetin treatment significantly up-regulated aliphatic glucosinolate biosynthesis genes compared with kaempferol and rutin. In addition, expression and metabolite analysis of the transparent testa7 mutant, which lacks hydroxylated flavonol derivatives, clarified the role of quercetin in the glucosinolate biosynthesis pathway. This study elucidates the molecular mechanisms by which flavonols interfere with signaling pathways, their molecular targets, and the multiple biological activities of flavonols in plants.

Gut Microbial and Associated Metabolite Markers for Colorectal Cancer Diagnosis.

Globally, colorectal cancer (CRC) is the second most common cause of mortality worldwide. Considerable evidence indicates that dysbiosis of the gut microbial community and its metabolite secretions play a fundamental role in advanced adenoma (ADA) and CRC development and progression. This study is a systematic review that aims to assess the clinical association between gut microbial markers and/or gut and circulating metabolites with ADA and CRC. Five electronic databases were searched by four independent reviewers. Only controlled trials that compared ADA and/or CRC with healthy control (HC) using either untargeted (16s rRNA gene or whole genome sequencing) or targeted (gene-based real-time PCR) identification methods for gut microbiome profile, or untargeted or targeted metabolite profiling approaches from the gut or serum/plasma, were eligible. Three independent reviewers evaluated the quality of the studies using the Cochrane Handbook for Systematic Reviews of Interventions. Twenty-four studies were eligible. We identified strong evidence of two microbial markers Fusobacterium and Porphyromonas for ADA vs. CRC, and nine microbial markers Lachnospiraceae-Lachnoclostridium, Ruminococcaceae-Ruminococcus, Parvimonas spp., Parvimonas micra, Enterobacteriaceae, Fusobacterium spp., Bacteroides, Peptostreptococcus-Peptostreptococcus stomatis, Clostridia spp.-Clostridium hylemonae, Clostridium symbiosum, and Porphyromonas-Porphyromonas asaccharolytica for CRC vs. HC. The remaining metabolite marker evidence between the various groups, including ADA vs. HC, ADA vs. HC, and CRC vs. HC, was not of sufficient quality to support additional findings. The identified gut microbial markers can be used in a panel for diagnosing ADA and/or CRC. Further research in the metabolite markers area is needed to evaluate the possibility to use in diagnostic or prognostic markers for colorectal cancer.

P22-041-23 A Study Protocol: Targeted Metabolite Profiling and the Effect of Maternal Lean Fish Intake During Pregnancy and Infancy

P22-041-23 A Study Protocol: Targeted Metabolite Profiling and the Effect of Maternal Lean Fish Intake During Pregnancy and Infancy

Determination of short-chain fatty acids as putative biomarkers of cancer diseases by modern analytical strategies and tools: a review.

Short-chain fatty acids (SCFAs) are the main metabolites produced by bacterial fermentation of non-digestible carbohydrates in the gastrointestinal tract. They can be seen as the major flow of carbon from the diet, through the microbiome to the host. SCFAs have been reported as important molecules responsible for the regulation of intestinal homeostasis. Moreover, these molecules have a significant impact on the immune system and are able to affect inflammation, cardiovascular diseases, diabetes type II, or oncological diseases. For this purpose, SCFAs could be used as putative biomarkers of various diseases, including cancer. A potential diagnostic value may be offered by analyzing SCFAs with the use of advanced analytical approaches such as gas chromatography (GC), liquid chromatography (LC), or capillary electrophoresis (CE) coupled with mass spectrometry (MS). The presented review summarizes the importance of analyzing SCFAs from clinical and analytical perspective. Current advances in the analysis of SCFAs focused on sample pretreatment, separation strategy, and detection methods are highlighted. Additionally, it also shows potential areas for the development of future diagnostic tools in oncology and other varieties of diseases based on targeted metabolite profiling.

Metabolomic and Transcriptomic Analysis Reveal the Role of Metabolites and Genes in Modulating Flower Color of Paphiopedilum micranthum.

Food-deceptive flowers primarily use visual signals (such as color) to mimic model plants and deceive insects into achieving pollination. Paphiopedilum micranthum is a food-deceptive orchid that has a pink labellum and two purple petals with a yellow base and has been proven to be pollinated by bumblebees. However, the chemical and molecular bases of the floral color are not well understood. We conducted targeted metabolite profiling and transcriptomic analysis to determine the color signal and its genetic basis in P. micranthum. We found that both anthocyanins and carotenoids contribute significantly to the formation of floral color that determines the color signal. Higher concentrations of anthocyanins (cyanidin and peonidin) and carotenoids (primarily lutein and zeaxanthin) were detected in the petal compared to the labellum. The upregulation of structural genes of CHS, F3'H, DFR and ANS on the anthocyanin biosynthesis pathway in petals was identified, as well as three genes of LCYE, BCH, and CCD4 on the carotenoid biosynthesis pathway. Furthermore, we discovered that three R2R3-MYBs and one bHLH transcription factors were co-expressed with the expression of different genes. These genes and transcription factors may be responsible for the spatial color difference of P. micranthum. Our study emphasizes that the color of this food-deceptive orchids is achieved through specific genes and transcription factors associated with the pigment biosynthesis pathway.

The associations between metabolic profiles and sexual and physical abuse in depressed adolescent psychiatric outpatients: an exploratory pilot study

ABSTRACT Background: Sexual and physical abuse have been associated with long-term systemic alterations such as low-grade inflammation and changes in brain morphology that may be reflected in the metabolome. However, data on the metabolic consequences of sexual and physical abuse remain scarce. Objective: This pilot study sought to investigate changes in the metabolite profile related to sexual and physical abuse in depressed adolescent psychiatric outpatients. Method: The study included 76 patients aged 14–18 years, whose serum samples were analysed with a targeted metabolite profiling methodology. We estimated the associations between metabolite concentrations and the Trauma and Distress Scale (TADS) Sexual and Physical Abuse factor scores using three linear regression models (one unadjusted and two adjusted) per metabolite and trauma type pair. Additional variables in the two adjusted models were 1) the lifestyle indicators body mass index, tobacco use, and alcohol use, and 2) depression scores and the chronicity of depression. Results: TADS Sexual Abuse scores associated positively with homogentisic acid, as well as cystathionine, and negatively with choline in linear regression analysis, whereas TADS Physical Abuse scores associated negatively with AMP, choline, γ-glutamyl cysteine and succinate, and positively with D-glucuronic acid. Conclusions: This pilot study did not include a healthy control group for comparison and the cohort was relatively small. Nevertheless, we observed alterations in metabolites related to one-carbon metabolism, mitochondrial dysfunction, oxidative stress, and inflammation in depressed patients with a history of sexual or physical abuse.

Mediterranean diet related metabolite profiles and cognitive performance

Evidence suggests that adherence to the Mediterranean diet (MedDiet) affects human metabolism and may contribute to better cognitive performance. However, the underlying mechanisms are not clear. We generated a metabolite profile for adherence to MedDiet and evaluated its cross-sectional association with aspects of cognitive performance. A total of 1250 healthy Greek middle-aged adults from the Epirus Health Study cohort were included in the analysis. Adherence to the MedDiet was assessed using the 14-point Mediterranean Diet Adherence Screener (MEDAS); cognition was measured using the Trail Making Test, the Verbal Fluency test and the Logical Memory test. A targeted metabolite profiling (n=250 metabolites) approach was applied, using a high-throughput nuclear magnetic resonance platform. We used elastic net regularized regressions, with a 10-fold cross-validation procedure, to identify a metabolite profile for MEDAS. We evaluated the associations of the identified metabolite profile and MEDAS with cognitive tests, using multivariable linear regression models. We identified a metabolite profile composed of 42 metabolites, mainly lipoprotein subclasses and fatty acids, significantly correlated with MedDiet adherence (Pearson r=0.35, P-value=5.5×10-37). After adjusting for known risk factors and accounting for multiple testing, the metabolite profile and MEDAS were not associated with the cognitive tests. A plasma metabolite profile related to better adherence to the MedDiet was not associated with the tested aspects of cognitive performance, in a middle-aged Mediterranean population.

Widely targeted metabolite profiling of mango stem apex during floral induction by compond of mepiquat chloride, prohexadione-calcium and uniconazole.

Insufficient low temperatures in winter and soil residues caused by paclobutrazol (PBZ) application pose a considerable challenge for mango floral induction (FI). Gibberellin inhibitors SPD (compound of mepiquat chloride, prohexadione-calcium and uniconazole) had a significant influence on enhancing the flowering rate and yield of mango for two consecutive years (2020-2021). Researchers have indicated that FI is regulated at the metabolic level; however, little is known about the metabolic changes during FI in response to SPD treatment. Here, ultra-performance liquid chromatography-electrospray ionization-tandem mass spectrometry (UPLC-ESI-MS/MS)-based widely targeted metabolomic analysis was carried out to assess the metabolic differences in the mango stem apex during different stage of mango FI (30, 80, 100 days after SPD/water treatment). A total of 582 compounds were annotated and 372 metabolites showed two-fold differences in abundance (variable importance in projection, VIP ≥ 1 and fold change, FC≥ 2 or≤ 0.5) between buds at 30, 80, 100 days after SPD/water treatment or between buds under different treatment. Lipids, phenolic acids, amino acids, carbohydrates, and vitamins were among metabolites showing significant differences over time after SPD treatment. Here, 18 out of 20 lipids, including the lysophosphatidylethanolamine (12, LPE), lysophosphatidylcholine (7, LPC), and free fatty acids (1, FA), were significantly upregulated from 80 to 100 days after SPD treatment comared to water treatment. Meanwhile, the dormancy release of mango buds from 80 to 100 days after SPD treatment was accompanied by the accumulation of proline, ascorbic acid, carbohydrates, and tannins. In addition, metabolites, such as L-homocysteine, L-histidine, and L-homomethionine, showed more than a ten-fold difference in relative abundance from 30 to 100 days after SPD treatment, however, there were no significant changes after water treatment. The present study reveals novel metabolites involved in mango FI in response to SPD, which would provide a theoretical basis for utilizing SPD to induce mango flowering.

Tumour catabolism independent of malnutrition and inflammation in upper GI cancer patients revealed by longitudinal metabolomics.

The detrimental impact of malnutrition and cachexia in cancer patients subjected to surgical resection is well established. However, how systemic and local metabolic alterations in cancer patients impact the serum metabolite signature, thereby leading to cancer-specific differences, is poorly defined. In order to implement metabolomics as a potential tool in clinical diagnostics and disease follow-up, targeted metabolite profiling based on quantitative measurements is essential. We hypothesized that the quantitative metabolic profile assessed by 1 H nuclear magnetic resonance (NMR) spectroscopy can be used to identify cancer-induced catabolism and potentially distinguish between specific tumour entities. Importantly, to prove tumour dependency and assess metabolic normalization, we additionally analysed the metabolome of patients' sera longitudinally post-surgery in order to assess metabolic normalization. Forty two metabolites in sera of patients with tumour entities known to cause malnutrition and cachexia, namely, upper gastrointestinal cancer and pancreatic cancer, as well as sera of healthy controls, were quantified by 1 H NMR spectroscopy. Comparing serum metabolites of patients with gastrointestinal cancer with healthy controls and pancreatic cancer patients, we identified at least 15 significantly changed metabolites in each comparison. Principal component and pathway analysis tools showed a catabolic signature in preoperative upper gastrointestinal cancer patients. The most specifically upregulated metabolite group in gastrointestinal cancer patients was ketone bodies (3-hydroxybutyrate, P<0.0001; acetoacetate, P<0.0001; acetone, P<0.0001; false discovery rate [FDR] adjusted). Increased glycerol levels (P<0.0001), increased concentration of the ketogenic amino acid lysine (P=0.03) and a significant correlation of 3-hydroxybutyrate levels with branched-chained amino acids (leucine, P=0.02; isoleucine, P=0.04 [FDR adjusted]) suggested that ketone body synthesis was driven by lipolysis and amino acid breakdown. Interestingly, the catabolic signature was independent of the body mass index, clinically assessed malnutrition using the nutritional risk screening score, and systemic inflammation assessed by CRP and leukocyte count. Longitudinal measurements and principal component analyses revealed a quick normalization of key metabolic alterations seven days post-surgery, including ketosis. Together, the quantitative metabolic profile obtained by 1 H NMR spectroscopy identified a tumour-induced catabolic signature specific to upper gastrointestinal cancer patients and enabled monitoring restoration of metabolic homeostasis after surgery. This approach was critical to identify the obtained metabolic profile as an upper gastrointestinal cancer-specific signature independent of malnutrition and inflammation.

Combination of DNA barcoding, targeted metabolite profiling and multispectral imaging to identify mold species and metabolites in sliced bread

Mould is a ubiquitous threat to safe food. In this work, a combination of three reliable methods to identify mould present in food at the species level is presented, to estimate the safety of the apparently non-moldy slices of bread and therefore contribute to the prevention of food waste. In overstored sliced bread, three mould species on the first slice of the stack (Chaetomium globosum, Penicillium chrysogenum and Scopulariopsis brevicaulis) were identified by DNA barcoding. The obtained sequences of the PCR products matched 100% in BLAST alignments with an e-value of 0. Targeted metabolite profiling confirmed the presence of the mycotoxin producers Chaetomium globosum and Penicillium chrysogenum. Known mycotoxins from these moulds (chaetoglobosin A and meleangrin, respectively) were quantitated (8.2 µg/g chaetoglobosin A and 2.2 µg/g meleangrin) in bread spot A. The second slice was less infected, which was revealed by only two mouldy spots, the lower amounts of determined mould metabolites and by normalized canonical discriminant analysis (nCDA) after multispectral imaging. On the third slice of the stack, no mould traces were detected by either metabolite profiling, or comparative multispectral imaging. These results suggest that it might be safe to consume bread slices in lower layers of bread stacks, even if mould growth is visible on the outer slices.