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Ajuga turkestanica preparations are used as anti-aging cosmeceuticals and for medicinal purposes. Herein we describe the characterization and quantification of its metabolites in different organs using UHPLC-MS and NMR spectroscopy. A total of 51 compounds belonging to various phytochemical classes (11 flavonoids, 10 ecdysteroids, 9 diterpenes, 6 fatty acids, 5 iridoids, 3 phenylpropanoids, 3 sugars, 2 phenolics, 1 coumarin, 1 triterpene) were annotated and tentatively identified by UHPLC-ESI-QqTOF-MS/MS of methanolic extracts obtained separately from the organs. 1D and 2D NMR spectroscopy independently confirmed the identity of six major compounds. The abundances of these main constituents in flowers, fruits, leaves, roots, seeds, and stems were compared and quantified using 1H NMR. The results showed that 8-O-acetylharpagide, 20-hydroxyecdysone (ecdysterone) and ajugachin B were the most abundant constituents in the species. The two major compounds, 8-O-acetylharpagide and 20-hydroxyecdysone, were chosen as the markers for the quality assessment of A. turkestanica material. The methanolic extract of the aerial parts of A. turkestanica showed no noteworthy anthelmintic (antihelmintic), antifungal, or cytotoxic effect in in vitro assays. [ABSTRACT FROM AUTHOR]

Centella asiatica, or pegagan in Indonesia, is a perennial plant used in Indonesian traditional medicine (jamu) and functional food with many biological activities. Those biological activities come from the bioactive metabolites present in C. asiatica. Differences in metabolite pathways in each part of the plant affect the accumulation of metabolites contained, thus impacting its biological activity. Therefore, this study aims to identify and evaluate differences in the distribution of metabolites in each part of C. asiatica, namely leaves, stems, stolons, and roots. Each plant part was extracted using methanol and sonicated for 30 minutes. The metabolites in the samples were separated and detected using UHPLC-QOrbitrap HRMS. Differences in the distribution of metabolites in each part of the plant were evaluated using chemometrics analysis. UHPLC-Q-Orbitrap HRMS analysis could positively identify 37 metabolites, most of which belong to the phenylpropanoid, triterpenoid, triterpenoid saponin, and flavonoid groups. Principal component analysis was able to clearly distinguish each part of the plant using the peak intensity of the overall chromatogram and the peak area of the identified metabolites. The different biosynthetic pathways of metabolites in plants could cause a difference in the distribution of metabolites in each plant. [ABSTRACT FROM AUTHOR]

Insects' host preferences are regulated by multiple factors whose interactions are only partly understood. Here we make use of an in-depth, untargeted metabolomic approach combining molecular networking (MN) and supervised Analysis of variance Multiblock Orthogonal Partial Least Squares (AMOPLS) to untangle egg-laying preferences of Drosophila suzukii, an invasive, highly polyphagous and destructive fruit pest originating from Southeast Asia. Based on behavioural experiments in the laboratory as well as field observation, we selected eight genetically related Vitis vinifera cultivars (e.g., Ancellotta, Galotta, Gamaret, Gamay, Gamay précoce, Garanoir, Mara and Reichensteiner) exhibiting significant differences in their susceptibility toward D. suzukii. The two most and the two least attractive red cultivars were chosen for further metabolomic analyses of their grape skins. The combination of MN and statistical AMOPLS findings with semi-quantitative detection information enabled us to identify flavonoids as interesting markers for differences in the attractiveness of the four studied grape cultivars towards D. suzukii. Overall, dihydroflavonols were accumulated in unattractive grape cultivars, while attractive grape cultivars were richer in flavonols. Crucially, both dihydroflavonols and flavonols were abundant metabolites in the semi-quantitative analysis of the extracted molecules from the grape skin. We discuss how these two flavonoid classes might influence the egglaying behaviour of D. suzukii females and how they could serve as potential markers for D. suzukii infestations in grapes that can be potentially extended to other fruits. We believe that our novel, integrated analytical approach could also be applied to the study of other biological relationships characterised by multiple evolving parameters. [ABSTRACT FROM AUTHOR]

Gastric cancer (GC) is the fifth most common cause of cancer-related death worldwide. Early detection is crucial for improving survival rates and treatment outcomes. However, accurate GC-specific biomarkers remain unknown. This study aimed to identify the metabolic differences between intestinal metaplasia (IM) and GC to determine the pathways involved in GC. A metabolic analysis of IM and tissue samples from 37 patients with GC was conducted using ultra-performance liquid chromatography with tandem mass spectrometry. Overall, 665 and 278 significant features were identified in the aqueous and 278 organic phases, respectively, using false discovery rate analysis, which controls the expected proportion of false positives among the significant results. sPLS-DA revealed a clear separation between IM and GC samples. Steroid hormone biosynthesis, tryptophan metabolism, purine metabolism, and arginine and proline metabolism were the most significantly altered pathways. The intensity of 11 metabolites, including N1, N2-diacetylspermine, creatine riboside, and N-formylkynurenine, showed significant elevation in more advanced GC. Based on pathway enrichment analysis and cancer stage-specific alterations, we identified six potential candidates as diagnostic biomarkers: aldosterone, N-formylkynurenine, guanosine triphosphate, arginine, S-adenosylmethioninamine, and creatine riboside. These metabolic differences between IM and GC provide valuable insights into gastric carcinogenesis. Further validation is needed to develop noninvasive diagnostic tools and targeted therapies to improve the outcomes of patients with GC. [ABSTRACT FROM AUTHOR]

Many naturally occurring chemical metabolites with significant cytotoxic activities have been isolated from medicinal plants and have become the leading hotspot of anti-cancer research in recent years. Hyptis rhomboidea Mart. et Gal is used as a folk medicine in South China to treat or assist in the treatment of liver disease, ulcers, and edema. But its chemical constituents have not been fully investigated yet. This study aimed to assess the cytotoxicity of H. rhomboidea, which was chemically characterized by chromatography–mass spectrometry methods. The results showed that the 95% ethanol extract of H. rhomboidea has marked inhibitory effects on five human cancer cell lines (HL-60, A549, SMMC-7721, MDA-MB-231, and SW480), with IC50 values ranging from 15.8 to 40.0 μg/mL. A total of 64 compounds were identified by ultra-high-performance liquid chromatography with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) and gas chromatograph–mass spectroscopy (GC-MS) analysis of H. rhomboidea crude extract. Among them, kaempferol, quercetin, rosmarinic acid, squalene, and campesterol were found to be abundant and might be the major metabolites involved to its bioactivity. The cytotoxic characterization and metabolite profiling of H. rhomboidea displayed in this research provides scientific evidence to support its use as medicinal properties. [ABSTRACT FROM AUTHOR]

Microalgae offer numerous benefits, including the production of value-added products like omega-3 fatty acids, pigments, and bioactive compounds, as well as their potential use as a third-generation biofuel. Euglena , a unicellular microalgae, is gaining attention as an industrially relevant organism and an emerging cell factory. It exhibits various trophic growth types depending on cultivation conditions. Light intensity and wavelength are critical factors affecting microalgal growth and productivity. This study explores the effects of different light-emitting diode (LED) wavelengths on the growth models and metabolite production of Euglena gracilis from peatland water. The highest specific growth rates were observed under blue and white light in both the logistic and Gompertz models. White light produced the highest biomass yield (0.813 ± 0.261 g.L−1). Our findings indicate that white light is the most effective treatment for enhancing growth and metabolite content, including lipids, carbohydrates, paramylon, and carotenoids. Conversely, green light resulted in the highest protein content. Different illumination treatments led to variations in lipid profiles and fatty acid composition. The highest SFA content was recorded under red light (59.6%), while the highest PUFA content was observed under blue light (31.1%), and the highest MUFA content was detected under red light (18.0%). In conclusion, this study highlights the impact of LEDs on the growth models of E. gracilis and their influence on the production of biomass, carbohydrates, proteins, chlorophyll, carotenoids, paramylon, and fatty acids. [ABSTRACT FROM AUTHOR]

Abstract Ajuga turkestanica preparations are used as anti-aging cosmeceuticals and for medicinal purposes. Herein we describe the characterization and quantification of its metabolites in different organs using UHPLC-MS and NMR spectroscopy. A total of 51 compounds belonging to various phytochemical classes (11 flavonoids, 10 ecdysteroids, 9 diterpenes, 6 fatty acids, 5 iridoids, 3 phenylpropanoids, 3 sugars, 2 phenolics, 1 coumarin, 1 triterpene) were annotated and tentatively identified by UHPLC-ESI-QqTOF-MS/MS of methanolic extracts obtained separately from the organs. 1D and 2D NMR spectroscopy independently confirmed the identity of six major compounds. The abundances of these main constituents in flowers, fruits, leaves, roots, seeds, and stems were compared and quantified using 1H NMR. The results showed that 8-O-acetylharpagide, 20-hydroxyecdysone (ecdysterone) and ajugachin B were the most abundant constituents in the species. The two major compounds, 8-O-acetylharpagide and 20-hydroxyecdysone, were chosen as the markers for the quality assessment of A. turkestanica material. The methanolic extract of the aerial parts of A. turkestanica showed no noteworthy anthelmintic (antihelmintic), antifungal, or cytotoxic effect in in vitro assays.

Bioactive compounds in functional foods, medicinal plants and others are considered attractive value-added molecules based on their wide range of bioactivity. It is clear that an important role is occupied by polyphenol, phenolic compounds and others. Urine is an effective biofluid to evaluate and monitor alterations in homeostasis and other processes related to metabolism. The current review provides a detailed description of the formation of urine in human body, various aspects relevant to sampling and analysis of urinary metabolites before presenting recent developments leveraging on metabolite profiling of urine. For the profiling of small molecules in urine, advancement of liquid chromatography mass tandem spectrometry (LC/MS/MS), establishment of standardized chemical fragmentation libraries, computational resources, data-analysis approaches with pattern recognition tools have made it an attractive option. The profiling of urinary metabolites gives an overview of the biomarkers associated with the diet and evaluates its biological effects. Metabolic pathways such as glycolysis, tricarboxylic acid cycle, amino acid metabolism, energy metabolism, purine metabolism and others can be evaluated. Finally, a combination of metabolite profiling with chemical standardization and bioassay in functional food and medicinal plants will likely lead to the identification of new biomarkers and novel biochemical insights. [ABSTRACT FROM AUTHOR]

Geosmithia pallida (KU693285) was isolated from Brucea mollis an endangered medicinal plant of North-East India. The secondary metabolites, produced by the endophytic fungi, extracted by ethyl acetate were screened for antimicrobial activity. G. pallida extract displayed the highest antimicrobial activity against Candida albicans with a minimum inhibitory concentration of 80.5 ± 1.25 µg/mL. G. pallida also showed the highest antioxidant activity which differed insignificantly from Penicillium sp. (P > 0.05). The G. pallida extract also exhibited the highest cellulase activity and also amylase and protease activities. The cytotoxicity assay of the ethyl acetate extract of this endophyte showed negligible effect (1.93 ± 0.42%) on chromosomal aberration as compared to the control (cyclophosphamide monohydrate) (7.20 ± 1.51%). The internal transcribed spacer rDNA sequence of G. pallida was submitted to the NCBI (Accession number KU693285) from India for the first time. The FT-IR spectrophotometry of the bioactive metabolite of G. pallida showed the presence of different functional groups such as alcohol, carboxylic acids, amines, aromatics, alkyl halides, aliphatic amines and alkynes. The GC-MS analysis revealed the presence of acetic acid, 2-phenylethyl ester; tetracosane; cyclooctasiloxane hexadecamethyl; cyclononasiloxane octadecamethyl; octadecanoic acid; phthalic acid, di(2-propylpentyl) ester and nonadecane, 2,6,10,14,18-pentamethyl as the major compounds in the metabolite. The findings of the present work indicated G. pallida as a potential source of important biomolecules without mammalian cytotoxic effects, which can be utilized for pharmaceutical purposes. [ABSTRACT FROM AUTHOR]

Tissue culture techniques were used to produce large amounts of bioactive compounds with medicinal potential, overcoming space and time constraints for cancer prevention. Rice callus suspension cultures (RCSC) and seed extracts prepared from aromatic rice varieties were used to evaluate the cytotoxic impact on human colon and lung cancer cell lines, as well as a normal control cell line, using Taxol as a positive control. RCSC and seed extracts from two Indian aromatic rice varieties were applied at different concentrations to treat the cancer cell lines and normal lung fibroblasts over varying time intervals. Apoptosis was assessed in 1:5 dilutions of the A549 and HT-29 cell lines treated with RCSC for 72 h, using propidium iodide staining and flow cytometry. RCSC showed a more potent cytotoxic effect than seed extracts with minimal effect on the normal cell line, in contrast to Taxol. Confocal microscopy and flow cytometry further confirmed the apoptotic effect of RCSC. Gas chromatography-mass spectrometry-based metabolic profiling identified metabolites involved in cytotoxicity and highlighted altered pathways. RCSC is proposed as an alternative source for the development of novel anticancer drugs with reduced side effects.

Abstract Functional foods, such as fermented bamboo shoots, have a long history of consumption among the ethnic communities in northeast India. These locally fermented bamboo shoots contain a wealth of beneficial microbes and metabolites that can help combat metabolic syndromes like obesity. However, the precise effects and mechanism behind fermented bamboo shoot products and their anti‐obesity properties remain unknown. This study aims to explore the different types of fermented bamboo shoot products to determine their potential anti‐obesity effects as well as to analyze their microbial diversity and metabolite profiles. Using both culture‐dependent and culture‐independent methods, we found a high abundance of lactic acid bacteria from the Firmicutes and Proteobacteria phyla in the sample. Gas chromatography–mass spectrometry (GC–MS) based untargeted metabolite profiling detected several aroma‐active compounds, bioactive metabolites, short‐chain fatty acids, and essential amino acids in the samples. The water extract derived from a particular type of fermented bamboo shoot, Melye‐amiley, was found to significantly reduce intracellular lipid accumulation in cultured 3T3‐L1 cells. In addition, this extract increased the expression of lipolytic (hormone‐sensitive lipase, lipoprotein lipase, and adipose triglyceride lipase) and browning regulator genes (uncoupling protein [UCP1], PRDM16, and PGC1‐alpha). By activating the AMPK signaling pathway, the water extract from Melye‐amiley also upregulated thermogenic protein expression and promoted mitochondrial biogenesis and fatty acid β‐oxidation. These findings suggest that fermented bamboo shoot extract has promising anti‐obesity effects by boosting energy expenditure in white adipocytes. Future research is necessary to identify the active ingredient(s) that may lead to new therapies to treat obesity.

An untargeted metabolomic analysis of five Eucalyptus species was used to compare their chemical profiles. Gas chromatography-mass spectrometry (GC-MS) along with multivariate analyses including principal component analysis (PCA) and hierarchical cluster analysis (HCA) were applied to assess variations in the leaf chemical profiles of E. maidenii F. Muell, E. robusta Smith, E. citriodora Hook (syn Corymbia citriodora), E. tereticornis Smith and E. camaldulensis Dehnh. In total, 299 relevant metabolites were identified and subjected to statistical analyses in order to detect either the major common metabolites of the genus or the specific compounds contributing to the characteristic chemical composition of each species. Organic acids, nitrogenous compounds, and phenolic constituents were the main discriminatory groups between the investigated eucalypts. The cluster analysis revealed that fatty acids and fatty alcohols were more concentrated in the leaves of E. citriodora and distinguished it from other species, whereas sugars, polyols, and glycosides were characteristic to E. camaldulensis.

More than 30 species of benthic Prorocentrum have been identified, some of which produce okadaic acid (OA) and its derivatives, dinophysistoxins (DTXs), which cause diarrhetic shellfish poisoning (DSP). Increasing numbers of benthic Prorocentrum species have been reported in tropical and subtropical waters of China. In contrast, only a few benthic Prorocentrum species have been reported in temperate waters. In this study, morphological descriptions obtained using light microscopy, scanning electron microscopy and molecular characterization of one Prorocentrum clipeus strain isolated from the Yellow Sea are presented. Prorocentrum clipeus cells were nearly circular in shape, with a collar, ridge, and one protrusion. The periflagellar area was wide U-shaped, with two curved projections on platelet 1a. Nine periflagellar platelets of different sizes were observed. The morphology closely fits that of the species isolated from other locations. Phylogenetic analysis based on the molecular sequences of the small subunit (SSU) rDNA, internal transcribed spacer (ITS), and large subunit (LSU) rDNA was performed. A comprehensive metabolomic analysis incorporating target, suspect and non-target screenings was first applied to investigate the intracellular and extracellular metabolite profiles of the current isolate of P. clipeus. According to the results of the target and suspect screenings, 179 metabolites or toxins produced by DSP-related algal species, including OA, dinophysistoxin-1 (DTX1), dinophysistoxin-2 (DTX2) and pectenotoxin-2 (PTX2), were not detected. Non-target screening involving feature-based molecular networking (FBMN) provided a global view of major metabolites produced by the P. clipeus DF128 strain and revealed 23 clusters belonging to at least 13 compound classes, with organometallic compounds, lipids and lipid-like molecules, phenylpropanoids and polyketides, and benzenoids as major types. To date, this is the first record of the characterization of P. clipeus in samples from Chinese waters. Our results support the wide distribution of epibenthic Prorocentrum species. [ABSTRACT FROM AUTHOR]

Metabolic perturbation has been associated with depression. An untargeted metabolomics approach using liquid chromatography-high resolution mass spectrometry was employed to detect and measure the rat serum metabolic changes following chronic social isolation (CSIS), an animal model of depression, and effective antidepressant fluoxetine (Flx) treatment. Univariate and multivariate statistics were used for metabolic data analysis and differentially expressed metabolites (DEMs) determination. Potential markers and predictive metabolites of CSIS-induced depressive-like behavior and Flx efficacy in CSIS were evaluated by the receiver operating characteristic (ROC) curve, and machine learning (ML) algorithms, such as support vector machine with linear kernel (SVM–LK) and random forest (RF). Upregulated choline following CSIS may represent a potential marker of depressive-like behavior. Succinate, stachydrine, guanidinoacetate, kynurenic acid, and 7-methylguanine were revealed as potential markers of effective Flx treatment in CSIS rats. RF yielded better accuracy than SVM–LK (98.50% vs. 85.70%, respectively) in predicting Flx efficacy in CSIS vs. CSIS, however, it performed almost identically in classifying CSIS vs. control (75.83% and 75%, respectively). Obtained DEMs combined with ROC curve and ML algorithms provide a research strategy for assessing potential markers or predictive metabolites for the designation or classification of stress-induced depressive phenotype and mode of drug action. [ABSTRACT FROM AUTHOR]

SUMMARY: Phytohormones are essential signaling molecules regulating various processes in growth, development, and stress responses. Genetic and molecular studies, especially using Arabidopsis thaliana (Arabidopsis), have discovered many important players involved in hormone perception, signal transduction, transport, and metabolism. Phytohormone signaling pathways are extensively interconnected with other endogenous and environmental stimuli. However, our knowledge of the huge and complex molecular network governed by a hormone remains limited. Here we report a global overview of downstream events of an abscisic acid (ABA) receptor, REGULATORY COMPONENTS OF ABA RECEPTOR (RCAR) 6 (also known as PYRABACTIN RESISTANCE 1 [PYR1]‐LIKE [PYL] 12), by integrating phosphoproteomic, proteomic and metabolite profiles. Our data suggest that the RCAR6 overexpression constitutively decreases the protein levels of its coreceptors, namely clade A protein phosphatases of type 2C, and activates sucrose non‐fermenting‐1 (SNF1)‐related protein kinase 1 (SnRK1) and SnRK2, the central regulators of energy and ABA signaling pathways. Furthermore, several enzymes in sugar metabolism were differentially phosphorylated and expressed in the RCAR6 line, and the metabolite profile revealed altered accumulations of several organic acids and amino acids. These results indicate that energy‐ and water‐saving mechanisms mediated by the SnRK1 and SnRK2 kinases, respectively, are under the control of the ABA receptor‐coreceptor complexes. Significance Statement: Phosphoproteomic, proteomic, and metabolite profiles of the Arabidopsis overexpressing an abscisic acid (ABA) receptor were integrated, providing a global overview of the complex network governed by a hormone. The overproduction of an ABA receptor caused the decreased protein levels of its coreceptor phosphatases, activating the central regulatory kinases of energy and stress signaling. [ABSTRACT FROM AUTHOR]

The flower buds of Zingiber striolatum Diels are considered a special vegetable in China, and they are rich in anthocyanins. However, the detailed composition types and the molecular mechanism of anthocyanin biosynthesis in Z. striolatum flower buds are still unclear. In this study, targeted metabolites were used to analyze and identify the anthocyanin types of Z. striolatum in three planting modes: monoculture (CK), intercropping with maize (ZP), and intercropping with soybean (SP). A total of 48 anthocyanins were identified with significant differential accumulation in Z. striolatum flower buds. Among them, cyanidin-3-O-glucoside was the main composition type of anthocyanins. Furthermore, the composition types of blue anthocyanin were identified in flower buds. A total of 15 structure genes were obtained from the transcriptome database of Z. striolatum flower buds. The qRT-PCR results revealed that the expression levels of ZsC4H-1, ZsC4H-2, ZsCHS-2, ZsCHI, ZsF3H, ZsF3′H, ZsDFR, ZsF3′5′H-3, and ZsANS genes were the highest in the ZP model. This study showed that the ZP model contributes to anthocyanin synthesis and accumulation of Z. striolatum flower buds among the three planting modes of Z. striolatum. These findings provide valuable information for research on the planting model and anthocyanin biosynthesis in Z. striolatum flower buds. [ABSTRACT FROM AUTHOR]

Cyanobacteria are photosynthetic prokaryotes with biologically active metabolites. The present work investigated the antimicrobial and antioxidant activities of extracts of six isolates (Cyanobacterium aponinum (0TAÇ3), Cylindrospermum moravicum (0TAÇ4), Nostoc sp. (0TAÇ5), Pseudoanabaena lonchoides (11TAÇ8), Cyanobium sp. (11TAÇ9), and Leptolyngbya sp. (11TAÇ13)) isolated from the Tersakan Stream. By applying the minimum inhibitory concentration (MIC) method, it was found that especially Pseudoanabaena lonchoides and Nostoc sp. extracts displayed strong antimicrobial activity (0.3125-10 mg/mL) on the tested microorganisms. 2,2-Diphenyl-1-picrylhydrazyl (DPPH) scavenging and Ferric reducing antioxidant power assay (FRAP) were investigated for antioxidant activity screening. Cyanobacterium aponinum and Cyanobium sp. extracts showed strong DPPH (IC50: 0.497 and 0.479 mg/mL, respectively) scavenging and Fe3+ reducing activities (16.88 and 13.58 mg GAE/g extract, respectively). Additionally, the total phenolic content was determined spectroscopically. The antioxidant activities of the methanol extracts were found to be related to the total phenolic content. Cyanobacterium aponinum and Leptolyngbya sp. extracts had high phenolic content (28.99 and 26.42 mg GAE/g extract, respectively). Metabolite profiling was performed by gas chromatography-mass spectroscopy (GC-MS) after the methanol extracts were derivatized with a silylation agent. The GC-MS analysis detected primary and secondary metabolites (fatty acids, organic and inorganic acids, hydrocarbons, alkaloids, terpenes, etc.) that might contribute to biological activity. According to these results, it was revealed that the studied cyanobacteria have therapeutic agents that exhibit natural antimicrobial and antioxidant activity. [ABSTRACT FROM AUTHOR]

Tissue culture techniques were used to produce large amounts of bioactive compounds with medicinal potential, overcoming space and time constraints for cancer prevention. Rice callus suspension cultures (RCSC) and seed extracts prepared from aromatic rice varieties were used to evaluate the cytotoxic impact on human colon and lung cancer cell lines, as well as a normal control cell line, using Taxol as a positive control. RCSC and seed extracts from two Indian aromatic rice varieties were applied at different concentrations to treat the cancer cell lines and normal lung fibroblasts over varying time intervals. Apoptosis was assessed in 1:5 dilutions of the A549 and HT-29 cell lines treated with RCSC for 72 h, using propidium iodide staining and flow cytometry. RCSC showed a more potent cytotoxic effect than seed extracts with minimal effect on the normal cell line, in contrast to Taxol. Confocal microscopy and flow cytometry further confirmed the apoptotic effect of RCSC. Gas chromatography-mass spectrometry-based metabolic profiling identified metabolites involved in cytotoxicity and highlighted altered pathways. RCSC is proposed as an alternative source for the development of novel anticancer drugs with reduced side effects. [ABSTRACT FROM AUTHOR]

Introduction: Changes in skin phenotypic characteristics are based on skin tissue. The study of the metabolic changes in skin tissue can help understand the causes of skin diseases and identify effective therapeutic interventions. Objectives: We aimed to establish and optimize a non-targeted skin metabolome extraction system for skin tissue metabolomics with high metabolite coverage, recovery, and reproducibility using gas chromatography/mass spectrometry. Methods: The metabolites in skin tissues were extracted using eleven different extraction systems, which were designed using reagents with different polarities based on sequential solid-liquid extraction employing a two-step strategy and analyzed using gas chromatograph/mass spectrometry. The extraction efficiency of diverse solvents was evaluated by coefficient of variation (CV), multivariate analysis, metabolites coverage, and relative peak area analysis. Results: We identified 119 metabolites and the metabolite profiles differed significantly between the eleven extraction systems. Metabolites with high abundances in the organic extraction systems, followed by aqueous extraction, were involved in the biosynthesis of unsaturated fatty acids, while metabolites with high abundances in the aqueous extraction systems, followed by organic extraction, were involved in amino sugar and nucleotide sugar metabolism, and glycerolipid metabolism. MeOH/chloroform-H2O and MeOH/H2O-chloroform were the extraction systems that yielded the highest number of metabolites, while MeOH/acetonitrile (ACN)-H2O and ACN/H2O-IPA exhibited superior metabolite recoveries. Conclusion: Our results demonstrated that our research facilitates the selection of an appropriate metabolite extraction approach based on the experimental purpose for the metabolomics study of skin tissue. [ABSTRACT FROM AUTHOR]

Perennial ryegrass (Lolium perenne L.) has potential in the phytoremediation of cadmium (Cd)-contaminated soil due to its strong Cd accumulation capacity and high biomass. In this study, we investigated the growth physiology, Cd accumulation, and metabolites of L. perenne roots under different soil acid stress levels (pH 4.0, 4.5, 5.0 and 6.0) and Cd concentrations (100 and 0 mg/kg) after 90 days of growth. The results showed that soil acid stress significantly impacts the remediation capability and physiological metabolic properties of L. perenne. Based on root Cd content and enrichment coefficient, soil pH between 4.5 and 5.0 was more conducive to Cd accumulation. The growth physiology and Cd accumulation of L. perenne were inhibited under high soil acid stress (pH 4.0). High soil acid stress caused a decrease in root length, root volume, and root biomass of L. perenne. Root malondialdehyde (MDA) content and the activity of antioxidant enzymes (catalase (CAT) and peroxidase (POD)) increased significantly in response to high soil acid stress to enhance tolerance. Metabolomics analysis revealed that acid stress resulted in significant changes in certain metabolites. Tartaric acid, fructose and amino acids (glutamate and lysine) in the roots of L. perenne were compatible solutes under acid stress. This study indicated that L. perenne has strong physiological and metabolic tolerance, as well as Cd accumulation ability, in response to soil acid stress. [ABSTRACT FROM AUTHOR]

Neural networks (NNs) are emerging as a rapid and scalable method for quantifying metabolites directly from nuclear magnetic resonance (NMR) spectra, but the nonlinear nature of NNs precludes understanding of how a model makes predictions. This study implements an explainable artificial intelligence algorithm called integrated gradients (IG) to elucidate which regions of input spectra are the most important for the quantification of specific analytes. The approach is first validated in simulated mixture spectra of eight aqueous metabolites and then investigated in experimentally acquired lipid spectra of a reference standard mixture and a murine hepatic extract. The IG method revealed that, like a human spectroscopist, NNs recognize and quantify analytes based on an analyte's respective resonance line-shapes, amplitudes, and frequencies. NNs can compensate for peak overlap and prioritize specific resonances most important for concentration determination. Further, we show how modifying a NN training dataset can affect how a model makes decisions, and we provide examples of how this approach can be used to de-bug issues with model performance. Overall, results show that the IG technique facilitates a visual and quantitative understanding of how model inputs relate to model outputs, potentially making NNs a more attractive option for targeted and automated NMR-based metabolomics. [ABSTRACT FROM AUTHOR]

Abstract Piper sarmentosum Roxb. (Piperaceae) is a traditional medicinal and food plant widely distributed in the tropical and subtropical regions of Asia, offering both health and culinary benefits. In this study the secondary metabolites in different organs of P. sarmentosum were identified and their relative abundances were characterized. The metabolic profiles of leaves, roots, stems and fruits were comprehensively investigated by liquid chromatography high-resolution mass spectrometry (LC-HR-MS) and the data subsequently analyzed using multivariate statistical methods. Manual interpretation of the tandem mass spectrometric (MS/MS) fragmentation patterns revealed the presence of 154 tentatively identified metabolites, mostly represented by alkaloids and flavonoids. Principle component analysis and hierarchical clustering indicated the predominant occurrence of flavonoids, lignans and phenyl propanoids in leaves, aporphines in stems, piperamides in fruits and lignan-amides in roots. Overall, this study provides extensive data on the metabolite composition of P. sarmentosum, supplying useful information for bioactive compounds discovery and patterns of their preferential biosynthesis or storage in specific organs. This can be used to optimize production and harvesting as well as to maximize the plant’s economic value as herbal medicine or in food applications. Graphical Abstract

Black tea's quality and flavor are largely influenced by its processing stages, which affect its volatile and non-volatile phytochemicals. This study aimed to optimized black tea manufacturing by investigating withering time, fermentation time, and temperature's impact on sensory quality. Using a U*15 (157) uniform design, optimal conditions were determined: 14 h of withering, 5.6 h of fermentation, and a 34 °C temperature. A verification experiment analyzed the volatile and non-volatile profiles. HPLC, GC–MS, and LC-MS revealed dynamic changes in phytochemicals. Among 157 VOCs and 2642 metabolites, 19 VOCs (VIP > 1.5) were crucial for aroma, while 50 (VIP > 1.5, p

In previous investigations, we have identified a rhizobacterium (Klebsiella sp. MBE02) that confers host protection against several phytopathogenic fungi. For instance, this rhizobacterium prevents Aspergillus flavus infection and promotes peanut growth and fitness in controlled and field-conditions. The mechanistic basis of the protective function offered by this rhizobacterium is not completely understood. MBE02 directly restricts the growth of the pathogenic fungi, which led us to hypothesize that it may strongly dysregulate the metabolism of A. flavus, and inhibit critical metabolic processes of the fungus, which severely restricts pathogen growth. We have tested this hypothesis by using untargeted metabolite profiling. Sixty-nine A. flavus metabolites accumulated differentially due to the presence of the MBE02. MBE02 could inhibit several important metabolic pathways, which include the biosynthesis of critical primary metabolites such as amino acids and fatty acids. It also impacts energy metabolism of the fungus, and that the accumulation of several structural components, including of the cell wall, were strongly inhibited. MBE02 abrogated the accumulation of disease-causing metabolites in A. flavus, whereas the accumulation of metabolites that inhibit fungal growth were enhanced. On the other hand, A. flavus did not strikingly impact the accumulation of metabolites of the MBE02. Our investigation supports the hypothesis that Klebsiella sp. MBE02 mediates protective function by directly impairing the pathogen’s metabolism.

This review explores the critical role of advanced metabolomic methodologies in interpreting the complex relationship between the bioactive compounds in olive oil and their health benefits. Olive oil, the cornerstone of the Mediterranean diet, is renowned for its numerous health benefits, including anti-inflammatory, antioxidant, and cardioprotective properties. This review begins with an overview of the importance of olive oil for nutrition and health, followed by a detailed discussion of the key metabolites that contribute to its therapeutic potential. It further explores the evolution of analytical techniques in metabolomics from traditional methods to cutting-edge technologies, such as mass spectrometry and nuclear magnetic resonance spectroscopy, which have enhanced our understanding of the complex composition of olive oil. This review highlights how these advanced analytical findings correlate with the known health properties of olive oil and presents case studies in which these methodologies have led to new insights. Finally, we address the challenges in standardizing these techniques and discuss future research directions, underscoring the significance of metabolomics in nutritional science and public health. This comprehensive analysis aims to provide a deeper understanding of the health benefits of olive oil, informed by the latest advancements in analytical methodologies. [ABSTRACT FROM AUTHOR]

• Phaleria macrocarpa is widely used as a remedy to reduce blood glucose levels. • GC–MS is a distinctive method dominating the metabolomics research area. • Chemometric analysis can be applied to resolve large data sets. Phaleria macrocarpa is a medicinal plant widely available in Malaysia. The plant parts have been traditionally used as an antidiabetic remedy. This study aimed to identify the putative inhibitors of the α-glucosidase enzyme from P. macrocarpa using a gas chromatography-mass spectrometry (GC–MS)-based metabolomics approach and further subjected them to in silico molecular docking analysis to elucidate the possible mechanism of action. This study assessed the inhibitory potential of P. macrocarpa fruit extracts (aqueous, 20 %, 40 %, 60 %, 80 %, and 100 % ethanol) against the α-glucosidase enzyme using GC–MS and chemometric analysis. Orthogonal partial least squares (OPLS) combined with GC–MS analysis were applied to correlate the inhibition of enzyme activity to the metabolites profile of P. macrocarpa. All the potential inhibitors identified were further docked to the yeast (Saccharomyces cerevisiae) protein crystal structure (PDB3A4A). The generated score scatter plot of OPLS showed a recognizable separation of all the extracts into six different clusters. GC–MS, incorporated with multivariate data analysis techniques, was used to identify significant chemical markers. Methyl α- d -glucopyranoside, squalene, palmitic acid, myo-inositol and isoquinoline metabolites were identified as putative inhibitors against α-glucosidase activity from P. macrocarpa. In conclusion, the GC–MS-based metabolomics approach identified potential chemical markers of P. macrocarpa that could be utilized in the development of herbal based medicine. [ABSTRACT FROM AUTHOR]

BACKGROUND: Symphytum (comfrey) genus, particularly Symphytum officinale, has been empirically used in folk medicine mainly for its potent anti‐inflammatory properties. In an attempt to shed light on the valorization of less known taxa, the current study evaluated the metabolite profile and antioxidant and enzyme inhibitory effects of nine Symphytum species. RESULTS: Phenolic acids, flavonoids and pyrrolizidine alkaloids were the most representative compounds in all comfrey samples. Hierarchical cluster analysis revealed that, within the roots, S. grandiflorum was slightly different from S. ibericum, S. caucasicum and the remaining species. Within the aerial parts, S. caucasicum and S. asperum differed from the other samples. All Symphytum species showed good antioxidant and enzyme inhibitory activities, as evaluated in DPPH (up to 50.17 mg Trolox equivalents (TE) g−1), ABTS (up to 49.92 mg TE g−1), cupric reducing antioxidant capacity (CUPRAC, up to 92.93 mg TE g−1), ferric reducing antioxidant power (FRAP, up to 53.63 mg TE g−1), acetylcholinesterase (AChE, up to 0.52 mg galanthamine equivalents (GALAE) g−1), butyrylcholinesterase (BChE, up to 0.96 mg GALAE g−1), tyrosinase (up to 13.58 mg kojic acid equivalents g−1) and glucosidase (up to 0.28 mmol acarbose equivalents g−1) tests. Pearson correlation analysis revealed potential links between danshensu and ABTS/FRAP/CUPRAC, quercetin‐O‐hexoside and DPPH/CUPRAC, or rabdosiin and anti‐BChE activity. CONCLUSIONS: By assessing for the first time in a comparative manner the phytochemical–biological profile of a considerably high number of Symphytum samples, this study unveils the potential use of less common comfrey species as novel phytopharmaceutical or agricultural raw materials. © 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. [ABSTRACT FROM AUTHOR]

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Recently, we could show that scallops show limitations of muscular performance like a reduced force under ocean warming and acidification. However, the underlying mechanisms at the cellular level are not completely understood. Metabolomics has become a valuable tool to evaluate the responses of marine organisms to various stressors. In the present study we therefore used a semi-targeted, multi tissue NMR based metabolomic approach to analyze metabolite patterns in the Atlantic king scallop, Pecten maximus, that were long-term acclimated to different end of century conditions of ocean warming (OW), ocean acidification (OA) and their combination (OWA). We investigated tissue specific metabolic profiles and metabolite concentrations in frozen tissues from gills, mantle and phasic and tonic adductor muscle of P. maximus under present conditions using ¹H-HR-MAS NMR spectroscopy. A set of 33 metabolites revealed a clear tissue-specific pattern which can be attributed to the individual functions of the respective tissue type. We then evaluated the impact of OW, OA and OWA on the metabolic profiles of the different tissues. OW was the main driver of the changes in metabolites. In particular, energy-related metabolites seem to play an important role in the physiological response of scallops to OW and OWA. In combination with pathway analysis and network exploration we propose a possible correlation between metabolic changes in the adductor muscle and limited swimming performance of P. maximus under future climate. While the metabolic response of the phasic muscle seems to mainly depend on net consumption of energy related metabolites such as ATP and phospho-L-arginine, the tonic muscle seems to rely on metabolizing specific amino acids and beta-oxidation to account for the elevated energetic requirements under ocean warming and acidification. [ABSTRACT FROM AUTHOR]

Dark–light and light–dark transitions during the day are switching points of leaf metabolism that strongly affect the regulatory state of the cells, and this change is hypothesized to affect the translatome. The cytosolic glyceraldehyde-3-phosphate dehydrogenases GAPC1 and GAPC2 function in glycolysis, and carbohydrate and energy metabolism, but GAPC1/C2 also shows moonlighting functions in gene expression and post-transcriptional regulation. In this study we examined the rapid reprogramming of the translatome that occurs within 10 min at the end of the night and the end of the day in wild-type (WT) Arabidopsis and a gapc1/c2 double-knockdown mutant. Metabolite profiling compared to the WT showed that gapc1/c2 knockdown led to increases in a set of metabolites at the start of day, particularly intermediates of the citric acid cycle and linked pathways. Differences in metabolite changes were also detected at the end of the day. Only small sets of transcripts changed in the total RNA pool; however, RNA-sequencing revealed major alterations in polysome-associated transcripts at the light-transition points. The most pronounced difference between the WT and gapc1/c2 was seen in the reorganization of the translatome at the start of the night. Our results are in line with the proposed hypothesis that GAPC1/C2 play a role in the control of the translatome during light/dark transitions. [ABSTRACT FROM AUTHOR]

Analysis was conducted to compare levodopa/carbidopa pharmacokinetics and drug‐related material in plasma of healthy participants after receiving a continuous infusion of Levodopa/Carbidopa Intestinal Gel (LCIG) to a continuous subcutaneous infusion of foslevodopa/foscarbidopa. Study samples were from a randomized, open‐label, 2‐period crossover study in 20 healthy participants. Participants received either 24‐h foslevodopa/foscarbidopa SC infusion to the abdomen or LCIG delivered for 24 h to the jejunum through a nasogastric tube with jejunal extension. Serial blood samples were collected for PK. Comparability of the LD PK parameters between the two treatment regimens was determined. Selected plasma samples were pooled per treatment group and per time point for metabolite profiling. LC–MSn was performed using high‐resolution mass spectrometry to identify drug‐related material across the dosing regimens and time points. The LD PK parameter central values and 90% confidence intervals following the foslevodopa/foscarbidopa subcutaneous infusion were between 0.8 and 1.25 relative to the LCIG infusion. With LCIG administration, LD, CD, 3‐OMD, DHPA, DOPAC, and vanillacetic acid were identified in plasma at early and late time points (0.75 and 24 h); the metabolic profile after administration of foslevodopa/foscarbidopa demonstrated the same drug‐related compounds with the exception of the administered foslevodopa. 3‐OMD and vanillacetic acid levels increased over time in both treatment regimens. Relative quantification of LC–MS peak areas showed no major differences in the metabolite profiles. These results indicate that neither the addition of monophosphate prodrug moieties nor SC administration affects the circulating metabolite profile of foslevodopa/foscarbidopa compared to LCIG. [ABSTRACT FROM AUTHOR]

In this study, flower and leaf extracts of Colvillea racemosa were considered a source of bioactive compounds. In this context, the objective of the study focused on investigating the anticancer potential as well as the phytochemical composition of both extracts. The extracts were analyzed by UPLC-ESI-QTOF-MS, and the bioactivity was tested using in vitro antioxidant assays (FRAP, DPPH, and ABTS) in addition to cytotoxic assays on non-small cell lung cancer cell line (A549). Our results clearly indicated the potent radical scavenging capacity of both extracts. Importantly, the flower extract exhibited a greater antioxidant capacity than the leaf extract. In terms of cytotoxic activity, leaf and flower extracts significantly inhibited cell viability with IC50 values of 17.0 and 17.2 µg/mL, respectively. The phytochemical characterization enabled the putative annotation of 42 metabolites, such as saccharides, phenolic acids, flavonoids, amino acids, and fatty acids. Among them, the flavonoid C-glycosides stand out due to their high relative abundance and previous reports on their anticancer bioactivity. For a better understanding of the bioactive mechanisms, four flavonoids (vitexin, kaempferol-3-O-rutinoside, luteolin, and isoorientin) were selected for molecular docking on hallmark protein targets in lung cancer as represented by γ-PI3K, EGFR, and CDK2 through in-silico studies. In these models, kaempferol-3-O-rutinoside and vitexin had the highest binding scores on γ-PI3K and CDK2, followed by isoorientin, so they could be highly responsible for the bioactive properties of C. racemosa extracts. [ABSTRACT FROM AUTHOR]

In this study, a chilli pepper (Capsicum annuum) panel for post-harvest carotenoid retention was studied to elucidate underlying mechanisms associated with this commercial trait of interest. Following drying and storage, some lines within the panel had an increase in carotenoids approaching 50% compared with the initial content at the fresh fruit stage. Other lines displayed a 25% loss of carotenoids. The quantitative determination of carotenoid pigments with concurrent cellular analysis indicated that in most cases, pepper fruit with thicker (up to 4-fold) lipid exocarp layers and smooth surfaces exhibit improved carotenoid retention properties. Total cutin monomer content increased in medium/high carotenoid retention fruits and subepidermal cutin deposits were responsible for the difference in exocarp thickness. Cutin biosynthesis and cuticle precursor transport genes were differentially expressed between medium/high and low carotenoid retention genotypes, and this supports the hypothesis that the fruit cuticle can contribute to carotenoid retention. Enzymatic degradation of the cuticle and cell wall suggests that in Capsicum the carotenoids (capsanthin and its esters) are embedded in the lipidic exocarp layer. This was not the case in tomato. Collectively, the data suggest that the fruit cuticle could provide an exploitable resource for the enhancement of fruit quality. [ABSTRACT FROM AUTHOR]

Phytohormones are essential signalling molecules globally regulating many processes of plants, including their growth, development, and stress responses. The promotion of growth and the enhancement of stress resistance have to be balanced, especially under adverse conditions such as drought stress, because of limited resources. Plants cope with drought stress via various strategies, including the transcriptional regulation of stress-responsive genes and the adjustment of metabolism, and phytohormones play roles in these processes. Although abscisic acid (ABA) is an important signal under drought, less attention has been paid to other phytohormones. In this review, we summarize progress in the understanding of phytohormone-regulated primary metabolism under water-limited conditions, especially in Arabidopsis thaliana , and highlight recent findings concerning the amino acids associated with ABA metabolism and signalling. We also discuss how phytohormones function antagonistically and synergistically in order to balance growth and stress responses. [ABSTRACT FROM AUTHOR]

The Calvin-Benson cycle (CBC) evolved over 2 billion years ago but has been subject to massive selection due to falling atmospheric carbon dioxide, rising atmospheric oxygen and changing nutrient and water availability. In addition, large groups of organisms have evolved carbon-concentrating mechanisms (CCMs) that operate upstream of the CBC. Most previous studies of CBC diversity focused on Rubisco kinetics and regulation. Quantitative metabolite profiling provides a top-down strategy to uncover inter-species diversity in CBC operation. CBC profiles were recently published for twenty species including terrestrial C 3 species, terrestrial C 4 species that operate a biochemical CCM, and cyanobacteria and green algae that operate different types of biophysical CCM. Distinctive profiles were found for species with different modes of photosynthesis, revealing that evolution of the various CCMs was accompanied by co-evolution of the CBC. Diversity was also found between species that share the same mode of photosynthesis, reflecting lineage-dependent diversity of the CBC. Connectivity analysis uncovers constraints due to pathway and thermodynamic topology, and reveals that cross-species diversity in the CBC is driven by changes in the balance between regulated enzymes and in the balance between the CBC and the light reactions or end-product synthesis. • Calvin-Benson cycle metabolites were profiled in 20 species. • Calvin-Benson cycle has co-evolved with CO 2 -concentrating mechanisms. • Substantial diversity between species with the same photosynthesis mode. [ABSTRACT FROM AUTHOR]

Erythrina bidwillii Lindl., Leguminosae, constitutes a valuable crop for horticulture and medicine; however, it is rarely investigated. Menopause is a crucial transitional period in women's health. Women worldwide consider the use of phytoestrogens as a safe hormone replacement therapy to alleviate detrimental menopausal symptoms. Thus, the discovery of novel phytoestrogens is highly demanded. The present study aimed to investigate, for the first time, the metabolomic profile and the estrogenic potential of E. bidwillii Lindl. leaf. Ultra‐performance liquid chromatography–electrospray ionization‐tandem mass spectrometry and gas chromatography–mass spectrometry metabolite profiling revealed the prevalence of alkaloids, flavonoids, isoflavonoids and fatty acids. Additionally, five erythrinan alkaloids, cristanine A (1), 8‐oxoerythraline (2), (+)‐erythrinine (3), (+)‐erythraline (4) and 8‐oxoerythrinine (5), along with the isoflavonoid genistin (6), were isolated. Erythrina bidwillii leaf extract exhibited significant in vivo estrogenic, anti‐osteoporotic, anti‐hyperlipidemic, hepatoprotective, and nephroprotective activities, utilizing ovariectomized rat model. Moreover, ethyl acetate and hexane fractions possessed significant in vitro estrogeic potential on MCF‐7 cell lines. An in silico study of the isolated metabolites revealed that (+)‐erythrinine (3) and 8‐oxoerythrinine (5) exhibited the highest affinity for ERα and ERβ, respectively, modeling them as potential estrogenic lead metabolites. Therefore, E. bidwillii leaf could be employed as promising hormone replacement therapy for postmenopausal women after thorough clinical trials. [ABSTRACT FROM AUTHOR]

The characterization of local improved varieties as well as the reduction of synthetic chemical fertilizers are sustainable approaches in the vision of a new precision Farming. Aim of our study was to improve the geographical characterization of local ecotypes and to identify peculiar features of new crops in terms of bioactive compounds. NMR and LC-MS metabolite profiling approaches followed by multivariate data analysis were applied to characterize local rosemary and garlic ecotypes. With the aim of applying for a protected designation of origin, orthogonal partial least squares discriminant analysis (OPLS-DA) was used to identify representative sensory quality indicators for Vessalico garlic and rosemary “Eretto Liguria” local ecotypes, Variable Influence on Projections (VIP) values of OPLS-DA indicated six metabolites as quality indicators for Vessalico garlic and sixteen metabolites as quality indicators for rosemary “Eretto Liguria”. Finally, to discover and utilize new ecotypes in a sustainable way, Vessalico garlic extracts antiviral activity, previously evaluated against Tomato brown rugose fruit virus (ToBRFV), a Tobamovirus affecting tomato crops, was extended to Pepino mosaic virus (PepMV) with positive results. [ABSTRACT FROM AUTHOR]

Insects’ host preferences are regulated by multiple factors whose interactions are only partly understood. Here we make use of an in-depth, untargeted metabolomic approach combining molecular networking (MN) and supervised Analysis of variance Multiblock Orthogonal Partial Least Squares (AMOPLS) to untangle egg-laying preferences of Drosophila suzukii, an invasive, highly polyphagous and destructive fruit pest originating from Southeast Asia. Based on behavioural experiments in the laboratory as well as field observation, we selected eight genetically related Vitis vinifera cultivars (e.g., Ancellotta, Galotta, Gamaret, Gamay, Gamay précoce, Garanoir, Mara and Reichensteiner) exhibiting significant differences in their susceptibility toward D. suzukii. The two most and the two least attractive red cultivars were chosen for further metabolomic analyses of their grape skins. The combination of MN and statistical AMOPLS findings with semi-quantitative detection information enabled us to identify flavonoids as interesting markers for differences in the attractiveness of the four studied grape cultivars towards D. suzukii. Overall, dihydroflavonols were accumulated in unattractive grape cultivars, while attractive grape cultivars were richer in flavonols. Crucially, both dihydroflavonols and flavonols were abundant metabolites in the semi-quantitative analysis of the extracted molecules from the grape skin. We discuss how these two flavonoid classes might influence the egg-laying behaviour of D. suzukii females and how they could serve as potential markers for D. suzukii infestations in grapes that can be potentially extended to other fruits. We believe that our novel, integrated analytical approach could also be applied to the study of other biological relationships characterised by multiple evolving parameters.

The utilization of metallic nanoparticles in bio-nanofabrication holds significant potential in the field of applied research. The current study applied and compared integrated ultrasonic-microwave-assisted extraction (US/MICE), ultrasonic extraction (USE), microwave-assisted extraction (MICE), and maceration (MAE) to extract total phenolic content (TPC). In addition, the study examined the antioxidant activity of Commiphora gileadensis (Cg) leaf. The results demonstrated that the TPC of US/MICE exhibited the maximum value at 59.34 ± 0.007 mg GAE/g DM. Furthermore, at a concentration of 10 μg/mL, TPC displayed a significant scavenging effect on DPPH (56.69 %), with an EC50 (6.48 μg/mL). Comprehensive metabolite profiling of the extract using UPLC-qTOF-MS/MS was performed to identify active agents. A total of 64 chromatographic peaks were found, out of which 60 were annotated. The most prevalent classes of metabolites found were polyphenols (including flavonoids and lignans), organic compounds and their derivatives, amides and amines, terpenes, and fatty acid derivatives. Transmission electron microscopy (TEM) revealed the aggregate size of the synthesized nanoparticles and the spherical shape of C. gileadensis-mediated silver nanoparticles (Cg-AgNPs). The nanoparticles had a particle size ranging from 7.7 to 42.9 nm. The Cg-AgNPs exhibited more inhibition zones against S. aureus and E. coli. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of Cg-extract, AgNPs, and Cg-AgNPs were also tested. This study demonstrated the feasibility of using combined ultrasonic-microwave-assisted extraction to separate and extract chemicals from C. gileadensis on a large scale. These compounds have potential use in the pharmaceutical industry. Combining antibacterial and biocompatible properties in materials is vital for designing new materials for biomedical applications. Additionally, the results showed that the biocompatibility of the Ag-NPs using C. gileadensis extracts demonstrated outstanding antibacterial properties.

Garcinia dulcis and Garcinia forbesii King are native plants from Indonesia and have tremendous potential as a source of raw medicines based on local wisdom. However, scientific data for strengthening pharmaceuticals are still limited. Therefore, it is necessary to conduct a study to strengthen and develop the potential of both plants using the approach of traditional medicine. This study aimed to explore the secondary metabolite composition and biological activity (antioxidant and antielastase) of both plants. Both samples were extracted using 70% ethanol and microwave-assisted extraction with a microwave power of 120 watts for 15 min. The extract obtained was then screened for phytochemicals using specific reagents. The total phenolic content (TPC) was determined using spectrophotometry with a 96-well microplate reader method. The total flavonoid content (TFC) was determined using the colorimetric method, whereas metabolite profiling analysis was conducted using the UPLC-QToF-MS/MS system. Meanwhile, biological activity was tested for antioxidant activity and antielastase as measured by a microplate reader 96-well spectrophotometry method at specific wavelengths. According to the results, G. dulcis and G. forbesii fruit peel extracts showed positive detection of particular secondary metabolites. TPC and TFC values were 13.98 ± 1.90 mg GAE/g and 10.33 ± 1.90 mg QE/g for G. dulcis and 11.98 ± 2.04 mgGAE/g and 1.96 ± 0.36 mgQE/g for G. forbesii. Metabolite profiling detected some compounds from G. dulcis, including ephedrannin B, hinokiflavone, mahuannin J, and candidate mass C9H12O8, and G. forbesii, including 5-Hydroxy-7,8,2'-trimethoxyflavone, lucialdehyde B, candidate mass C21H39NO4, candidate mass C14H10O6, and candidate mass C14H12O6. Meanwhile, the biological activities (antioxidant and antielastase) were 137.721 μg/mL and 108.893 μg/mL for G. dulcis and 481.948 μg/mL and 250.611 μg/mL for G. forbesii, respectively. Both plants showed different profiles of secondary metabolites and biological activities (antioxidant and antielastase) according to their respective characteristics.

Abstract The evaluation of plant-based feedstocks is an important aspect of biorefining. Nicotiana glauca is a solanaceous, non-food crop that produces large amounts of biomass and is well adapted to grow in suboptimal conditions. In the present article, compatible sequential solvent extractions were applied to N. glauca leaves to enable the generation of enriched extracts containing higher metabolite content comparing to direct leaf extracts. Typically, between 60 to 100 metabolite components were identified within the fractions. The occurrence of plant fatty acids, fatty acid alcohols, alkanes, sterols and terpenoids was detected by gas liquid chromatography–mass spectrometry (GC–MS) and metabolite identification was confirmed by comparison of physico-chemical properties displayed by available authentic standards. Collectively, co-products such waxes, oils, fermentable sugars, and terpenoids were all identified and quantified. The enriched fractions of N. glauca revealed a high level of readily extractable hydrocarbons, oils and high value co-products. In addition, the saccharification yield and cell wall composition analyses in the stems revealed the potential of the residue material as a promising lignocellulosic substrate for the production of fermentable sugars. In conclusion a multifractional cascade for valuable compounds/commodities has been development, that uses N. glauca biomass. These data have enabled the evaluation of N. glauca material as a potential feedstock for biorefining. Graphical Abstract

ABSTRACTThe present study determined total phenolic content (TPC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, α-glucosidase, and nitric oxide (NO) inhibitory activities of black (BR), purple (PR), and red rice (RR) extracted with different polarity solvents and characterized the metabolites by using the 1H NMR-based metabolomics approach. The methanol extract of RR had the highest DPPH scavenging and α-glucosidase inhibitory activities. The water extracts of PR and RR exhibited the highest NO inhibitory activity with IC50 values of 176.28 ± 18.78 and 160.69 ± 14.83 µg/mL, respectively. The metabolites that contribute to radical scavenging and α-glucosidase inhibitory activities of RR methanol extract were chlorogenic acid, protocatechuic acid, and tyrosine, whereas nicotinic acid, γ-aminobutyric acid, and citric acid were the metabolites that contributed to the NO inhibitory activity of RR water extract. The RR extracts are potential sources of bioactive compounds with antioxidant and NO inhibitory properties.

Gastric cancer (GC) is the fifth most common cause of cancer-related death worldwide. Early detection is crucial for improving survival rates and treatment outcomes. However, accurate GC-specific biomarkers remain unknown. This study aimed to identify the metabolic differences between intestinal metaplasia (IM) and GC to determine the pathways involved in GC. A metabolic analysis of IM and tissue samples from 37 patients with GC was conducted using ultra-performance liquid chromatography with tandem mass spectrometry. Overall, 665 and 278 significant features were identified in the aqueous and 278 organic phases, respectively, using false discovery rate analysis, which controls the expected proportion of false positives among the significant results. sPLS-DA revealed a clear separation between IM and GC samples. Steroid hormone biosynthesis, tryptophan metabolism, purine metabolism, and arginine and proline metabolism were the most significantly altered pathways. The intensity of 11 metabolites, including N1, N2-diacetylspermine, creatine riboside, and N-formylkynurenine, showed significant elevation in more advanced GC. Based on pathway enrichment analysis and cancer stage-specific alterations, we identified six potential candidates as diagnostic biomarkers: aldosterone, N-formylkynurenine, guanosine triphosphate, arginine, S-adenosylmethioninamine, and creatine riboside. These metabolic differences between IM and GC provide valuable insights into gastric carcinogenesis. Further validation is needed to develop noninvasive diagnostic tools and targeted therapies to improve the outcomes of patients with GC.

Many naturally occurring chemical metabolites with significant cytotoxic activities have been isolated from medicinal plants and have become the leading hotspot of anti-cancer research in recent years. Hyptis rhomboidea Mart. et Gal is used as a folk medicine in South China to treat or assist in the treatment of liver disease, ulcers, and edema. But its chemical constituents have not been fully investigated yet. This study aimed to assess the cytotoxicity of H. rhomboidea, which was chemically characterized by chromatography–mass spectrometry methods. The results showed that the 95% ethanol extract of H. rhomboidea has marked inhibitory effects on five human cancer cell lines (HL-60, A549, SMMC-7721, MDA-MB-231, and SW480), with IC50 values ranging from 15.8 to 40.0 μg/mL. A total of 64 compounds were identified by ultra-high-performance liquid chromatography with quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) and gas chromatograph–mass spectroscopy (GC-MS) analysis of H. rhomboidea crude extract. Among them, kaempferol, quercetin, rosmarinic acid, squalene, and campesterol were found to be abundant and might be the major metabolites involved to its bioactivity. The cytotoxic characterization and metabolite profiling of H. rhomboidea displayed in this research provides scientific evidence to support its use as medicinal properties.

Harsh climates, i.e., drought, extreme temperatures, and toxic gases, pose issues to agriculture by altering plants' growth and yield. Biotechnology with biochemical defense approaches is beneficial for generating new plants/varieties with extra resilience to adverse conditions. In response to stress, cultures show an enriched level of secondary metabolite synthesis. Here, an efficient in vitro propagation method using axillary shoot proliferation, along with callus formation, was established in Glycyrrhiza glabra L. The phytochemical composition of in vitro and in vivo grown tissues was analyzed using a gas chromatography–mass spectrometry (GC–MS) technique, and the biochemical attributes were measured and compared in different investigated tissues. Callus formation from root explants was achieved with a frequency of 88.89% on MS medium containing 2.0 mg/L BAP and 0.5 mg/L 2,4-D. Axillary shoot proliferation was obtained from dormant buds when cultured onto MS supplemented with BAP alone, or in combination with, IAA. The maximum shoot proliferation (94.44%) was recorded on MS with 1.0 mg/L BAP with an average shoot length of 10.5 cm. The regenerated shoots were subcultured and transferred to the root induction medium, supplemented with various concentrations of IAA/IBA, wherein 2.0 mg/L IBA resulted in the best rooting frequency (88.89%). The GC–MS-based phytocompounds analysis of the methanolic extracts of root-derived callus and in vivo- and in vitro- grown root tissues was conducted. These samples revealed the presence of more than 35 therapeutically important bioactive compounds, such as methylglabridin, sitosterol, lupeol, squalene, stearic acid, linoleic acid, etc. The biochemical parameters, like total phenolic content, flavonoid content, DPPH scavenging activity, superoxide dismutase, and peroxidase activity were also measured. All the biochemical attributes were found to be higher in in vitro derived roots than the callus and in vivo grown root (donor) samples. These findings demonstrated that callus (root derived) and in vitro roots are a stable and potent source of multiple phytocompounds, encompassing medical significance with wide applications. This study may serve as an alternative opportunity in the sustained and continuous synthesis of important compounds without harming natural vegetation and normal environment. [ABSTRACT FROM AUTHOR]

The discovery of new natural products has become more challenging because of the re-isolation of compounds and the lack of new sources. Microbes dwelling in extreme conditions of high salinity and temperature are huge prospects for interesting natural metabolites. In this study, the endophytic bacteria Bacillus velezensis 7NPB-3B isolated from the halophyte Salicornia brachiata was screened for its biofilm inhibition against methicillin-resistant Staphylococcus aureus (MRSA). The fractionation of the crude extract was guided by bioassay and LC-HRMS-based metabolomics using multivariate analysis. The 37 fractions obtained by high-throughput chromatography were dereplicated using an in-house MS-Excel macro coupled with the Dictionary of Natural Products database. Successive bioactivity-guided separation yielded one novel compound (1), a diketopiperazine (m/z 469.258 [M − H]−) with an attached saturated decanoic acid chain, and four known compounds (2–5). The compounds were identified based on 1D- and 2D-NMR and mass spectrometry. Compounds 1 and 5 exhibited excellent biofilm inhibition properties of >90% against the MRSA pathogen at minimum inhibition concentrations of 25 and 35 µg/mL, respectively. The investigation resulted in the isolation of a novel diketopiperazine from a bacterial endophyte of an untapped plant using an omics approach. [ABSTRACT FROM AUTHOR]

Objective: YZJ-4729 is a novel G protein-biased μ-opioid receptor agonist for the treatment of acute pain in adult patients who require intravenous opioid analgesic therapy. The aim of this study was to assess the pharmacokinetics, metabolite profiling, safety and tolerability of YZJ-4729 in healthy Chinese subjects following the single intravenous doses ranged from 0.2 mg to 6 mg. Methods: This single-center, randomized, double-blind, placebo-controlled clinical study was conducted in 54 healthy male and female Chinese subjects after single ascending doses of YZJ-4729 tartrate (0.2, 0.5, 1.5, 3, 4.5, and 6 mg). Subjects in each cohort were assigned randomly to receive a single intravenous dose of YZJ-4729 tartrate injection or placebo at a ratio of 4:1. Pharmacokinetic characteristics, metabolite profiling, safety and tolerability profiles of the study drug were evaluated. Results: Overall, YZJ-4729 was safe and well tolerated in healthy Chinese subjects. The study drug reached peak plasma concentrations nearly at the end of the infusion. After administration, YZJ-4729 was eliminated rapidly with a terminal elimination half-life of 0.862-2.50 h, and excreted little in human excreta. The maximum drug concentration and area under the plasma concentration-time curve increased with dose escalation across the entire dose range. YZJ-4729 experienced extensive metabolism in human body. A total of 19 metabolites were identified and the characteristic metabolic pathways involved hydroxylation, ketone formation, N-dealkylation and glucuronide conjugation. Metabolite M10 was the most abundant circulating metabolite, and represented over 10% of total drug-related systemic exposure. Further PK and safety evaluation of M10 was necessary. Conclusion: The clinical study results laid a foundation for the further clinical studies of YZJ-4729 in patients. [ABSTRACT FROM AUTHOR]