Potential Metabolites Research Articles

Effect of Glycoconjugation on Cytotoxicity and Selectivity of 8-Aminoquinoline Derivatives Compared to 8-Hydroxyquinoline.

Numerous emerging chemotherapeutic agents incorporate N-heterocyclic fragments in their structures, with the quinoline skeleton being particularly significant. Our recent works have focused on glycoconjugates of 8-hydroxyquinoline (8-HQ), which demonstrated enhanced bioavailability and solubility compared to their parent compounds, although they fell short in selectivity. In this study, our objective was to improve the selectivity of glycoconjugates by replacing the oxygen atom with nitrogen by substituting the 8-HQ moiety with 8-aminoquinoline (8-AQ). The 8-AQ derivatives were functionalized through the amino group and linked to sugar derivatives (D-glucose or D-galactose) that were modified with an azide, alkylazide, or propargyl group at the anomeric position by copper(I)-catalyzed 1,3-dipolar azido-alkyne cycloaddition (CuAAC). The resulting glycoconjugates, as well as their potential metabolites, were evaluated for their ability to inhibit the proliferation of cancer cell lines (including HCT 116 and MCF-7) and a healthy cell line (NHDF-Neo). Two of the synthesized glycoconjugates (17 and 18) demonstrated higher cytotoxicity than their oxygen-containing counterparts and showed improved selectivity for cancer cells, thus enhancing their anticancer potential. Furthermore, it was found that glycoconjugates exhibited greater cytotoxicity in comparison to their potential metabolites.

Unveiling molecular DOM reactomics and transformation coupled with multifunctional nanocomposites under anaerobic conditions: Tracking potential metabolomics and pathways.

Anaerobic digestion (AD) offers great potential for pollutant removal and bioenergy recovery. However, it faces challenges when using livestock manure (LSM) as a feedstock given its high content of refractory materials (e.g., lignocellulose, long-chain carbohydrates, lipids, and crude protein). This would significantly inhibit AD-microbial activities, reduce organic transformation efficiency and limit gas production. To overcome this, multifunctional metal-doped hydrochars (HCs) were introduced here as AD supplements/accelerators, given that LSM degradation under AD results in complex dissolved organic matter (DOM). To assess this, the current study investigates the molecular interactions/transformations within DOM during LSM-AD coupled with metal-doped HCs, via batch-mode experiments. Expansive data mining techniques were employed to analyze DOM using Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS). Substantial increments in peptide-like along with decrements in highly unsaturated-like molecules were observed in HC@MnCl2 containing-system. This indicates an increased capability for substrate hydrolysis and potential utilization of soluble microbial products (SMPs) (i.e., highly unsaturated-like molecules), leading to enhanced methane recovery (223.23 mL/g-VSadded, 1.77 times more than the control). However, accumulation of DOM-highly unsaturated molecules (i.e., a lack of SMPs' degradation) accompanied with low methane production (39.68 mL/g-VSadded) was noticed for HC@NiFe2O4. DOM reactivity during LSM-AD was validated via paired mass difference molecular network, indicating predominance of CHO and N-containing groups' transformations for HC@MnCl2 and HC@NiFe2O4, respectively. Potential metabolites and abundant pathways were verified via KEGG database. This study improves our understanding of LSM-AD-DOM complex transformation matrix, the fate of bioavailable/recalcitrant compounds, and identification of potential DOM regulators from thousands of molecules.

Quantifying liver-toxic responses from dose-dependent chemical exposures using a rat genome-scale metabolic model.

Because the liver plays a vital role in the clearance of exogenous chemical compounds, it is susceptible to chemical-induced toxicity. Animal-based testing is routinely used to assess the hepatotoxic potential of chemicals. While large-scale high-throughput sequencing data can indicate the genes affected by chemical exposures, we need system-level approaches to interpret these changes. To this end, we developed an updated rat genome-scale metabolic model to integrate large-scale transcriptomics data and utilized a chemical structure similarity-based ToxProfiler tool to identify chemicals that bind to specific toxicity targets to understand the mechanisms of toxicity. We used high-throughput transcriptomics data from a 5-day in vivo study where rats were exposed to different non-toxic and hepatotoxic chemicals at increasing concentrations and investigated how liver metabolism was differentially altered between the non-toxic and hepatotoxic chemical exposures. Our analysis indicated that the genes identified via toxicity target analysis and those mapped to the metabolic model showed a distinct gene expression pattern, with the majority showing upregulation for hepatotoxicants compared to non-toxic chemicals. Similarly, when we mapped the metabolic genes at the pathway level, we identified several pathways in carbohydrate, amino acid, and lipid metabolism that were significantly upregulated for hepatotoxic chemicals. Furthermore, using our system-level integration of gene expression data with the rat metabolic model, we could differentiate metabolites in these pathways that were systematically elevated or suppressed due to hepatotoxic versus non-toxic chemicals. Thus, using our combined approach, we were able to identify a set of potential gene signatures that clearly differentiated liver toxic responses from non-toxic chemicals, which helped us identify potential metabolic pathways and metabolites that are systematically associated with the toxicant exposure.

New acorane-sesequiterpenes and anti-retinoblastoma constituents from the marine algicolous fungus Trichoderma harzianum NTU2180 guided by molecular networking strategy

BackgroundTrichoderma species, known as biocontrol agents against plant diseases, contain diverse compounds, especially terpenoids, with various bioactivities. To facilitate the exploration of bioactive secondary metabolites of Trichoderma harzianum NTU2180, the OSMAC approach MS/MS molecular networking was applied in the current study.ResultsThe feature-based molecular networking (FBMN) analysis showed that T. harzianum NTU2180 fermented on germinated brown rice (GBR) produced more terpenoids. Here, two new acorane-sesequiterpenes, trichospirols A (1) and B (2), and 12 known compounds (3 − 14) were isolated from the EtOAc layer of T. harzianum NTU2180 fermentation on GBR. Structures of these compounds were determined through NMR, UV, IR, and MS analyses. The absolute configuration of trichospirols A (1) was also elucidated by x-ray with Cu K-α radiation. Among them, six compounds (1, 2, 3, 4, 5, and 11) were annotated as terpenoids by the NPClassifier on FBMN. 5-Hydroxy-3-hydroxmethyl-2-methyl-7-methoxychromone (7) and ergosterol peroxide (11) showed significant anti-angiogenic activity in ex vivo experiments with respective 0.57 ± 0.12- and 0.20 ± 0.12-fold changes. In addition, compound 11 displayed cytotoxicity against Y79 retinoblastoma cells with IC50 value of 35.3 ± 6.9 µM.ConclusionsThe current study utilizes FBMN concept with OSMAC approach to accelerate the exploration of potential metabolites of the fungus Trichoderma harzianum NTU2180. Through a series of FBMN-guided isolation and purification, two new acorane-sesequiterpenes and 12 known compounds were obtained. The ex vivo and in vitro experiments were evaluated to assess anticancer isolates. It is worth noting that compound 11 was identified as a dual inhibitor targeting both angiogenesis and proliferation of retinoblastomas. Altogether, the results revealed the novel potential of T. harzianum for developing natural therapeutics against retinoblastomas.

Untargeted Characterization and Biological Activity of Amazonian Aqueous Stem Bark Extracts by Liquid and Gas Chromatography-Mass Spectrometry.

Aqueous stem bark extracts of Aspidosperma rigidum Rusby, Couroupita guianensis Aubl., Monteverdia laevis (Reissek) Biral, and Protium sagotianum Marchand have been reported as traditional remedies in several countries of the Amazonian region. Despite previous research, further investigation to characterize secondary metabolites and the biological activity of extracts is needed to derive potential applications. Metabolic profiling was carried out using liquid and gas chromatography coupled with mass spectrometry (UHPLC-MS/MS and GC-MS). The chemical composition of the studied plants was further compared by principal component analysis (PCA). Additionally, chemical profiles were correlated with antimicrobial and toxicity activities, which suggested potential metabolites for future research. We identified 16 and 32 compounds by UHPLC-MS/MS and GC-MS analysis, respectively. Antimicrobial activity was detected in three stem bark extracts. C. guianensis showed inhibition of all tested microorganisms, including antibiotic-resistant strains. Molecular networking approaches, in silico tools, and Pearson's correlation showed that antifungal compounds could be a terpene glycoside (r = 0.918) and/or a phenolic (r = 0.882) metabolite class. This study highlights the use of the established procedure in exploring the metabolomes of these species, which could be a novel source of antimicrobial drug discovery. Coupling the observed biological potential with UHPLC-MS/MS data has also accelerated the tracing of their bioactive compounds. These findings update the state of the art regarding the chemical composition and biological activity of the plant extracts, defining potential new applications for the pharmaceutical applications.

Specific plasma metabolite profile in intestinal Behçet’s syndrome

BackgroundIntestinal Behçet’s syndrome (IBS) has high morbidity and mortality rates with serious complications. However, there are few specific biomarkers for IBS. The purposes of this study were to investigate the distinctive metabolic changes in plasma samples between IBS patients and healthy people, active IBS and inactive IBS patients, and to identify candidate metabolic biomarkers which would be useful for diagnosing and predicting IBS.MethodsIn this study, we performed a global untargeted metabolomics approach in plasma samples from 30 IBS patients and 20 healthy subjects. P value < 0.05 and variable importance projection (VIP) values > 1 were considered to be statistically significant metabolites. Univariate receiver operating characteristic (ROC) curve analysis was plotted as a measure for assessing the clinical performance of metabolites, and area under curve (AUC) were assessed.ResultsA total of 147 differentially abundant metabolites (DAMs) were identified between IBS patients and normal control (NC) group. The potential pathways involved in the pathogenesis of IBS include linoleic acid metabolism; GABAergic synapse; biosynthesis of unsaturated fatty acids; valine, leucine and isoleucine biosynthesis; ovarian steroidogenesis; and others. In addition, a total of 103 significant metabolites were selected to distinguish active IBS from inactive IBS patients. Tyrosine metabolism, dopaminergic synapse and neuroactive ligand-receptor interaction were found to be closely related to the disease activity of IBS. Furthermore, three potential metabolites including quinate, stearidonic acid (SDA) and capric acid (CA) could significantly differ IBS patients from NC group. On the other hand, 1-methyladenosine (m1A), genipin, methylmalonic acid (MMA) and ascorbate could significantly differentiated active IBS from inactive IBS patients.ConclusionIn conclusion, this study demonstrated the characteristic plasma metabolic profiles between IBS group and NC group, as well as between active and inactive IBS patients by using an untargeted LC/MS metabolomics profiling approach. In this study, quinate, SDA and CA were identified as potential diagnostic biomarkers for IBS. Additionally, m1A, genipin, MMA and ascorbate could serve as potential biomarkers for evaluating IBS activity. These findings might provide potential valuable insights for developing therapeutic strategies to manage IBS in the future.

Metabolome and RNA-seq reveal discrepant metabolism and secretory metabolism profile in skeletal muscle between obese and lean pigs at different ages.

Metabolites and metabolism-related gene expression profiles in skeletal muscle change dramatically under obesity, aging and metabolic disease. Since obese and lean pigs are ideal models for metabolic research. Here, we compared metabolome and transcriptome of Longissimus dorsi (LD) muscle between Taoyuan black (TB, obese) and Duroc (lean) pigs at different ages. We defined the "window phase" of intramuscular fat (IMF) deposition in TB pig, which has significantly higher IMF than Duroc pig. Our results displayed discrepant lipid composition and different expression genes (DEGs) enriched in lipid metabolism, and both metabolome and transcriptome analyses revealed stronger energy expenditure and more active amino acid and protein metabolism in Duroc pig. 10 up- and 51 down-regulated biomarker metabolites with age- and breed-specificity were identified. Potential secretory metabolites, including organic acid (fumaric acid, succinate, malic acid, and gamma-aminobutyric acid), amino acid (L-lysine, and L-glutamic acid), lipid (2-hydroxyisovaleric acid, and L-carnitine) were demonstrated a significant correlation with IMF deposition. Our research highlights the huge difference of metabolic spectrum in skeletal muscle between obese and lean model and muscle-derived secretory metabolites might act as an ambassador of intercellular communication to regulate systematic metabolism.

Untargeted Metabolomics Reveals the Metabolic Characteristics and Biomarkers of Antioxidant Properties of Gardeniae Fructus from Different Geographical Origins in China.

Background/Objectives: Gardeniae Fructus (GF) has been widely used as both food and medicinal purposes for thousands of years, but their antioxidant properties and potential metabolite biomarkers remain unclear. Methods: The purposes of this study were to examine antioxidant activities of 21 GF varieties from different geographical origins in China and identify potential biomarkers of antioxidant properties using an untargeted LC-MS-based metabolomics approach. Results: The results demonstrate that metabolomics had the ability to trace the geographical origins of GF. We found that antioxidant activities varied with different varieties of GF, which was dependent on their chemical compositions. The key chemical categories were obtained as the primary contributors of the antioxidant activity, including prenol lipids, flavonoids, coumarins and derivatives, as well as steroids and steroid derivatives. In addition, adouetine Y, coagulin R 3-glucoside and epicatechin 3-glucoside were identified as potential biomarkers for the antioxidant activity of GF. Conclusions: Therefore, our study sheds light on the metabolic characteristics and biomarkers of the antioxidant properties of GF, contributing to the selection and cultivation of a high antioxidant variety.

Xuefu Zhuyu Decoction Improves Blood-Brain Barrier Integrity in Acute Traumatic Brain Injury Rats via Regulating Adenosine.

To explore the neuroprotective effects of Xuefu Zhuyu Decoction (XFZYD) based on in vivo and metabolomics experiments. Traumatic brain injury (TBI) was induced via a controlled cortical impact (CCI) method. Thirty rats were randomly divided into 3 groups (10 for each): sham, CCI and XFZYD groups (9 g/kg). The administration was performed by intragastric administration for 3 days. Neurological functions tests, histology staining, coagulation and haemorheology assays, and Western blot were examined. Untargeted metabolomics was employed to identify metabolites. The key metabolite was validated by enzyme-linked immunosorbent assay and immunofluorescence. XFZYD significantly alleviated neurological dysfunction in CCI model rats (P<0.01) but had no impact on coagulation function. As evidenced by Evans blue and IgG staining, XFZYD effectively prevented blood-brain barrier (BBB) disruption (P<0.05, P<0.01). Moreover, XFZYD not only increased the expression of collagen IV, occludin and zona occludens 1 but also decreased matrix metalloproteinase-9 (MMP-9) and cyclooxygenase-2 (COX-2), which protected BBB integrity (all P<0.05). Nine potential metabolites were identified, and all of them were reversed by XFZYD. Adenosine was the most significantly altered metabolite related to BBB repair. XFZYD significantly reduced the level of equilibrative nucleoside transporter 2 (ENT2) and increased adenosine (P<0.01), which may improve BBB integrity. XFZYD ameliorates BBB disruption after TBI by decreasing the levels of MMP-9 and COX-2. Through further exploration via metabolomics, we found that XFZYD may exert a protective effect on BBB by regulating adenosine metabolism via ENT2.

Discovery of highly active kynureninases for cancer immunotherapy through protein language model.

Tailor-made enzymes empower a wide range of versatile applications, although searching for the desirable enzymes often requires high throughput screening and thus poses significant challenges. In this study, we employed homology searches and protein language models to discover and prioritize enzymes by their kinetic parameters. We aimed to discover kynureninases as a potentially versatile therapeutic enzyme, which hydrolyses L-kynurenine, a potent immunosuppressive metabolite, to overcome the immunosuppressive tumor microenvironment in anticancer therapy. Subsequently, we experimentally validated the efficacy of four top-ranked kynureninases under in vitro and in vivo conditions. Our findings revealed a catalytically most active one with a nearly twofold increase in turnover number over the prior best and a 3.4-fold reduction in tumor weight in mouse model comparisons. Consequently, our approach holds promise for the targeted quantitative enzyme discovery and selection suitable for specific applications with higher accuracy, significantly broadening the scope of enzyme utilization. A web-executable version of our workflow is available at seekrank.steineggerlab.com and our code is available as free open-source software at github.com/steineggerlab/SeekRank.

Exploring the mediating role of blood metabolites in the relationship between gut microbiota and gastric cancer risk: a Mendelian randomization study.

Prior studies have established correlations between gut microbiota (GM) dysbiosis, circulating metabolite alterations, and gastric cancer (GC) risk. However, the causal nature of these associations remains uncertain. We utilized summary data from genome-wide association studies (GWAS) on GM (European, n=8,956), blood metabolites (European, n=120,241; East Asian, n=4,435), and GC (European, n=476,116; East Asian, n=167,122) to perform a bidirectional Mendelian randomization (MR) analysis, investigating the causal effects of GM and metabolites on GC risk. Additionally, we conducted mediation analysis (two-step MR) to identify potential metabolite mediators in the GM-GC relationship. We identified twelve negative and seven positive associations between specific GM taxa and GC risk. For blood metabolites, seven traits were found to be significantly associated with reduced GC risk in the European population, with these findings subsequently validated in the East Asian cohort. Three GM taxa showed potential causal associations with five metabolic traits: the Bacteroidia class and Bacteroidales order were positively correlated with five metabolites (all P < 0.013), while Bacteroides OTU97_27 exhibited a negative correlation with one metabolite (P = 0.007). Two-step MR analysis indicated that total lipids in intermediate-density lipoprotein (IDL), IDL particle concentration, phospholipids in medium low-density lipoprotein (LDL), phospholipids in small LDL, and free cholesterol in small LDL indirectly influenced the association between Bacteroidia class/Bacteroidales order and GC, with mediation proportions of 1.71% (P = 0.048), 1.69% (P = 0.048), 2.05% (P = 0.045), 1.85% (P = 0.048), and 1.99% (P = 0.045), respectively. The present study provides suggestive evidence of a causal relationship between specific GM, blood metabolites, and GC risk, potentially offering new insights into GC etiology.

Metabolomic profiling and antibacterial efficacy of probiotic-derived cell-free supernatant encapsulated in nanostructured lipid carriers against canine multidrug-resistant bacteria.

This study aimed to investigate the antibacterial efficacy of probiotic-derived cell-free supernatants (CFS) encapsulated within nanostructured lipid carriers (NLCs) against multidrug-resistant Pseudomonas aeruginosa and Staphylococcus pseudintermedius. Additionally, it aimed to identify specific bioactive compounds that contribute to the reported antibacterial properties by characterizing the metabolite substances present in the CFS using a metabolomic analysis technique. Eight strains of lactic acid bacteria including Lactiplantibacillus plantarum (L22F and L25F), Pediococcus acidilactici (P72N, BF9, BF 14, BYF 20 and BYF 26) and Ligilactobacillus salivarius (BF 12) were selected as probiotic candidates. The inhibitory activity of their cell free supernatant (CFS) was tested against clinical strains of P. aeruginosa and S. pseudintermedius isolated from skin wounds of dogs and cats. An untargeted metabolomic approach based on liquid chromatography-mass spectrometry (LC-MS) identified potential antibacterial metabolites in the CFS. Cell-Free Supernatants-Nanostructured Lipid Carriers (CFS-NLCs) were developed, and their antibacterial activity and minimum bactericidal concentration (MBC) were analysed. Despite the strong multidrug-resistant nature of the pathogens, CFS displayed a moderate antibacterial activity against most tested strains. The acidic nature of the CFS, combined with bioactive antibacterial metabolites like Kanzonol V and 1-Hexanol, likely contributed to its inhibitory effects against pathogenic bacteria; notably, Kanzonol V was abundant in the CFS of L22F, BF12 and BYF26 (L22F_CFS, BF12_CFS and BYF26_CFS), while 1-Hexanol was particularly enriched in CFS of P72N (P72N_CFS), with both compounds effectively targeting bacterial cell membranes to disrupt cell integrity, leading to bacterial cell death. Other beneficial compounds such as Pyroglutamylleucine, Trigoneoside VIII and 18-Nor-4(19),8,11,13-abietatetraene which are likely to have anti-inflammatory, antimicrobial and antioxidant activities, were also detected in the CFS. The CFS-NLCs maintained their antibacterial activity and 30-60% dilutions of product completely inhibited the growth of pathogen strains even after three-months storage at room temperature. These findings suggest that CFS-NLCs could be a promising biotic therapy for treating hospital infections such as canine dermatitis and otitis caused by multidrug-resistant P. aeruginosa and S. pseudintermedius.

Development of a liquid chromatography-mass spectrometry based targeted metabolomics method for discovering diagnostic biomarkers in Kawasaki disease.

Kawasaki disease (KD) has emerged as the leading cause of acquired heart disease in children, primarily due to the absence of highly sensitive and specific biomarkers for early and accurate diagnosis. To address this issue, a simple and comprehensive targeted metabolomics method employing ultra high-performance liquid chromatography coupled with Q-TRAP mass spectrometry has been developed to identify new metabolite biomarkers for KD. This method enables the simultaneous quantification of 276 metabolites, covering 60 metabolic pathways, with a particular emphasis on metabolites relevant to KD. The use of nine ISs and commercial quality control samples significantly enhances both accuracy and precision. Through validation and application to serum samples from patients with KD, seventeen differential serum metabolites were identified. The altered metabolites are primarily associated with three functional metabolic pathways: tricarboxylic acid cycle, tryptophan metabolism, and bile acid metabolism, all of which are believed to be involved in the inflammatory and immune responses in KD patients. Ultimately, eight differential metabolites (indole-3-propionic acid, thiamine, indolepyruvic acid, levodopa, l-selenomethionine, isocitric acid, trans-aconitate, and N-acetylasparagine) were identified that could potentially serve as diagnostic biomarkers with the area under the curve values exceeding 0.9. Our targeted metabolomics approach demonstrates applicability in identifying potential metabolite biomarkers for KD and holds great promise in unraveling the intricate pathophysiology of the disease.

Malate initiates a proton-sensing pathway essential for pH regulation of inflammation

Metabolites can double as a signaling modality that initiates physiological adaptations. Metabolism, a chemical language encoding biological information, has been recognized as a powerful principle directing inflammatory responses. Cytosolic pH is a regulator of inflammatory response in macrophages. Here, we found that L-malate exerts anti-inflammatory effect via BiP-IRF2BP2 signaling, which is a sensor of cytosolic pH in macrophages. First, L-malate, a TCA intermediate upregulated in pro-inflammatory macrophages, was identified as a potent anti-inflammatory metabolite through initial screening. Subsequent screening with DARTS and MS led to the isolation of L-malate-BiP binding. Further screening through protein‒protein interaction microarrays identified a L-malate-restrained coupling of BiP with IRF2BP2, a known anti-inflammatory protein. Interestingly, pH reduction, which promotes carboxyl protonation of L-malate, facilitates L-malate and carboxylate analogues such as succinate to bind BiP, and disrupt BiP-IRF2BP2 interaction in a carboxyl-dependent manner. Both L-malate and acidification inhibit BiP-IRF2BP2 interaction, and protect IRF2BP2 from BiP-driven degradation in macrophages. Furthermore, both in vitro and in vivo, BiP-IRF2BP2 signal is required for effects of both L-malate and pH on inflammatory responses. These findings reveal a previously unrecognized, proton/carboxylate dual sensing pathway wherein pH and L-malate regulate inflammatory responses, indicating the role of certain carboxylate metabolites as adaptors in the proton biosensing by interactions between macromolecules.

OligoDistiller: A Platform Agnostic Software Tool for MS and MS2 Data Analysis of Complex Oligonucleotides and Their Impurities.

Oligonucleotides are currently one of the most rapidly advancing classes of therapeutic modalities. Understanding critical quality attributes, such as the impurity profile, stability, potential metabolites, and sequence conformity, is the key to their ultimate success. To obtain the information presented above, liquid chromatography-mass spectrometry (LC-MS) is often employed. However, data interpretation can be challenging due to multiple charge states, unknown species, chemical noises, and overlapping isotope envelopes (OIEs) of distinct but unresolved species. To address these challenges, we have developed an MS-platform agnostic, multifunctional R package and a web-tool for automated and interactive MS and MS2 data analysis, OligoDistiller. From the MS spectrum of a complex mixture of two synthetic strands, our tool OligoDistiller annotated and quantified 45 oligonucleotide-related features including 13 unknown impurities and 6 OIEs, which all together explained 90.8% of the detected peaks. Also, major product ions were assigned from the MS2 spectrum of a 47-mer DNA strand, covering 80.3% of the oligonucleotide sequence. We provide not only diverse isotope quality metrics for each annotated feature but also an interactive data review module allowing for direct inspection of the part of raw spectrum linked to a selected feature. This tool OligoDistiller is freely available at https://github.com/daniellyz/OligoDistiller.

AVALIAÇÃO DA ATIVIDADE HEPATOPROTETORA DO EXTRATO AQUOSO DAS FOLHAS DE CHRYSOBALANUS ICACO

The metabolization of substances in the human body is carried out mainly by the liver, but some substances when in excess, such as Paracetamol, can lead to liver toxicity. Chrysobalanus icaco L., a plant with potent antioxidant metabolites, shows promise in combating antioxidant imbalance and hepatoprotection action. The objective of this study was to evaluate the protective action of the aqueous extract of C. icaco leaves (EAFCi) against paracetamol-induced hepatotoxicity. For this, the plant material was collected and the leaf extract was prepared through decoction in distilled water (5:100 w/v). For the in vivo evaluation of the hepatoprotective activity, Wistar rats were divided into 3 groups of 5 animals each, which were treated with saline, silymarin and EAFCi at a dose of 100 mg/kg (groups I, II and III, respectively). On the seventh day of treatment, Paracetamol was administered and, after twelve days, euthanasia was performed to remove the liver. Reduced glutathione levels, lipid peroxidation, and histopathological examination were then evaluated. With the methodology used, a reduction in MDA levels and an increase in GSH levels were observed in the groups that were administered silymarin and the 100mg/kg dose of EAFCi, when compared to the control group. In addition, in the histological analysis of the liver, it was observed that the animals treated with silymarin and with the dose of 100mg/kg of EAFCi had normal liver morphology, unlike the negative control group, which showed significant changes. In this sense, it is concluded that the administration of EAFCi at a dose of 100mg/kg in Wistar rats was able to inhibit the hepatotoxicity induced by Paracetamol by decreasing MDA, increasing GSH and preserving liver morphology.

Whole genome sequence and LC-Mass for identifying antimicrobial metabolites of Bacillus licheniformis endophyte

Antimicrobial resistance (AMR) represents a critical public health issue that requiring immediate action. Wild halophytic plants can be the solution for the AMR crisis because they harbor unique endophytes capable of producing potent antimicrobial metabolites. This study aimed at identifying promising and antimicrobial metabolites produced by endophytic/epiphytic bacteria recovered from the wild Bassia scoparia plant. Standard methods were employed for the isolation of endophytes/epiphytes. Whole genome sequence (WGS) using Oxford Nanopore technology followed by antiSMASH analysis coupled with advanced LC-MS spectroscopic analysis were used for identification of the active antimicrobial metabolites. This study identified Bacillus licheniformis strain CCASU-B18 as a promising endophytic bacterium from the Bassia scoparia plant. In addition, the strain showed broad-spectrum antibacterial activity against three standard and five MDR clinical Gram-positive and Gram-negative isolates, and antifungal activity against the standard C. albicans strain. Six main antimicrobial metabolites—thermoactinoamide A, bacillibactins, lichenysins, lichenicidins, fengycin, and bacillomycin—were verified to exist by whole genome sequencing for identifying the respective conserved biosynthetic gene clusters in conjunction with LC/MS-MS analysis. The complete genomic DNA (4125835) and associated plasmid (205548 bp) of the promising endophytic isolate were sequenced, assembled, annotated, and submitted into the NCBI GenBank database under the accession codes, CP157373. In conclusion, Bacillus licheniformis strain CCASU-B18, a promising endophytic bacterium exhibiting broad-spectrum antimicrobial activities, was isolated. Future research is highly recommended to optimize the culture conditions that will be employed to enhance the production of respective antimicrobial metabolites, as well as testing these compounds against a broader range of MDR-resistant pathogens.

ANTI-INFLAMMATORY AND ANTICANCER POTENTIAL OF SALSOLA KALI L. AGAINST SKIN CANCER IN ALBINO RAT MODEL

In the realm of cancer treatment, plants are offering effective curative compounds and most of the anticancer compounds have been obtained from the plants. Salsola kali L. is an important ethnomedicinal plant of the Chenopodiaceae family. In folkloric healthcare practices, it is being used to treat skin diseases. In the current study, plant extracts were prepared in solvents of different polarities through soxhlation and subjected to the detection of potential bioactive metabolites through Fourier Transform Infrared Spectroscopy and Gas Chromatography-Mass Spectrometry. The anti-inflammatory efficacy of plant extract was also determined through carrageenan and histamine assays. The anticancer potential of the plant was tested against the cancerous skin of albino mice. The studied plant had encouraging anti-inflammatory efficacy. The carrageenan percent inhibition occurred in a dose-response manner and peak edema volume reduction occurred at 4 hr. Findings of the carrageenan assay were also confirmed with the histamine assay. At 120 min maximum edema inhibition was detected and inflammation was reduced upto 65%. Animal study on rats, presented S. kali L. as a potential local biological resource for skin cancer treatment. Chemotherapy of animals with n-Hexane extract showed excellent results in curing benign lesions. Squamous cell carcinoma in situ and malignant fibrous hyperplasia were cured mildly, and malignancy progression did not occur. Plant extract’s anticancer activity also coincided with the presence of potent bioactive compounds that were recorded in IR-spectra; triterpenoid and glycosidic saponins as O-H, C=O, C-H and C=C and C-O groups, respectively. Ethyl acetate also had good efficacy against the skin lesions. However, methanol extract showed less outcomes. GC-MS analysis of n-Hexane extract revealed the presence of an anticancer epoxide Cis-2.3-Epoxycotane. Ethyl acetate extract also showed the presence of an anticancer carbohydrate 1,6-Dideoxy-1-mannitol. These two anticancer compounds have been reported in the literature as having cancer cure potential. Further, after cytotoxic analysis studied plant may be proven as a potential chemotherapeutic resource for skin cancer treatment.

Girard Derivatization-Based Enrichment Strategy for Profiling the Carbonyl Submetabolome in Biological Samples.

Numerous bioactive compounds containing carbonyl groups, including aldehydes and ketones, are widely acknowledged as potential biomarkers for several diseases and are implicated in the development of metabolic disorders. However, the detection of carbonyl metabolites is hindered by challenges, such as poor ionization efficiency, low biological concentration, instability, and complexity of the sample matrix. To overcome these limitations, we developed a Girard derivatization-based enrichment (GDBE) strategy for capturing and comprehensively profiling carbonyl metabolites in biological samples. A functionalized resin, named carbonyl capture and reporter-ion installation (CCRI) resins, was synthesized to selectively capture carbonyl metabolites via a Girard reaction. After unwanted metabolites were removed, the hydrazone derivatives were cleaved from the solid-phase resins and subjected to LC-MS analysis. The proposed GDBE strategy exhibits exceptional selectivity for capturing and enriching carbonyl metabolites. Moreover, this method surpasses current detection limits by enhancing the MS sensitivity and facilitating structural characterization of hydrazone derivatives by a specific MS/MS fragmentation signature. Using the GDBE method, 957 potential carbonyl metabolites were successfully identified in liver tissue from alcohol-fed mice. Among them, 76 carbonyl metabolites were annotated, indicating the potential of this strategy for the efficient nontargeted profiling of the carbonyl submetabolome in complex biological samples.

Gas Chromatography-Tandem Mass Spectrometry Method for Analyses of Iodoform and Diiodomethane in Multiple Biological Matrices from Dairy Cattle.

A quantitative method was developed and validated to analyze iodoform and its potential metabolite, diiodomethane, in biological fluids from dairy cows, including rumen fluid, duodenal fluid, blood serum, milk, and urine, using liquid-liquid extraction (LLE) and GC-MS/MS. The method showed no matrix effects across different samples, recoveries of spiked samples between 70 and 120%, and relative standard deviations (RSD%) ranging from 0.7 to 14%. Inter- and intravariations in quality control samples were within 3.1-12.3%, with the highest variation noted in iodoform spiking. Normalized internal standard (IS) and matrix factor (MF) calculations confirmed the suitability of diiodomethane-d2 as an internal standard for quantifying both compounds across different matrices. These results validate the method's application for ADME (absorption, distribution, metabolism, and excretion) studies, enabling the investigation of iodoform digestion, metabolism, and excretion in dairy cattle.