Indoleacrylic Acid Research Articles

Biosynthesis, characterization, and in-vitro anticancer effect of plant-mediated silver nanoparticles using Acalypha indica Linn: In-silico approach

Cancer is a significant global health issue, with rising prevalence and mortality rates demanding urgent attention. The World Health Organization emphasizes the need for effective prevention, early detection, and treatment strategies to address this public health challenge. Current treatment modalities, including surgery, hormonal therapy, immunotherapy, radiation therapy, and chemotherapy, are often associated with considerable side effects and high costs. This study investigates the biosynthesis of silver nanoparticles using Acalypha indica L. (AgNPs), a medicinal plant recognized for its therapeutic benefits, as a potential cancer treatment with minimal side effects and a lower risk of drug resistance. AgNPs exhibit anti-inflammatory properties and the ability to inhibit angiogenesis while counteracting drug resistance mechanisms. Moreover, the use of chitosan as a coating on AgNPs (AgNPs-Chit) enhances their stability and specificity toward cancer cells, thereby improving their anticancer efficacy. Characterization of the synthesized AgNPs was conducted using various techniques, including UV-Vis spectroscopy, X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), particle size analysis (PSA), and transmission electron microscopy (TEM), confirming the successful synthesis, stability, and spherical morphology of the nanoparticles, with an average diameter of 7 nm and a zeta potential of −24.51 mV. In vitro cytotoxicity testing showed that AgNPs-Chit exhibited stronger anticancer activity against T47D cells with an IC50 value of 173 µg/mL, compared to AgNPs (IC50 244 µg/mL) and the Acalypha indica L extract (IC50 826 µg/mL). When compared to the control, treatments with AgNPs-Chit, AgNPs, and the plant extract demonstrated statistically significant differences (∗p < 0.05, ∗∗p < 0.01). These results indicate that the modification of AgNPs with chitosan (AgNPs-Chit) significantly enhances anticancer efficacy compared to both AgNPs and Acalypha indica L. extract. The modification with AgNPs increased anticancer efficiency by 338%, while AgNPs-Chit showed a 446% increase compared to the original extract, highlighting the enhanced potential of these nanoparticles in inhibiting cancer cell growth. Additionally, molecular docking studies of eight key compounds identified through LC-MS analysis (quercetin, kaempferol, catechin, indoline, 4-aminobenzoic acid, 1-(2-quinolinyl)piperazine, 3-indoleacrylic acid, and pyridine-3-carboxamide) revealed strong binding affinities to the cancer target protein 3PP0, with binding energies ranging from −9.4 to −5.9 kcal/mol, compared to doxorubicin's binding energy of −9.0 kcal/mol.

Gut microbiota-derived tryptophan metabolites regulated by Wuji Wan to attenuate colitis through AhR signaling activation

Disruption of the intestinal mucosal barrier caused by gut dysbiosis and metabolic imbalance is the underlying pathology of inflammatory bowel disease (IBD). Traditional Chinese medicine Wuji Wan (WJW) is commonly used to treat digestive system disorders and showed therapeutic potential for IBD. In this interdisciplinary study, we aim to investigate the pharmacological effects of WJW against experimental colitis by combining functional metabolomics and gut-microbiota sequencing techniques. Treatment with WJW altered the profile of the intestinal microbiota and notably increased the abundance of Lactobacillus, thereby facilitating the conversion of tryptophan into indole-3-acetic acid (IAA) and indoleacrylic acid (IA). These indole derivatives activated the aryl hydrocarbon receptor (AhR) pathway, which reduced colonic inflammation and restored the expression of intestinal barrier proteins. Interestingly, the beneficial effects of WJW on gut barrier function improvement and tryptophan metabolism were disappeared in the absence of gut microbiota. Finally, pre-treatment with the AhR antagonist CH-223191 confirmed the essential role of IAA-mediated AhR activation in the therapeutic effects of WJW. Overall, WJW enhanced intestinal barrier function and reduced colonic inflammation in a murine colitis model by modulating Lactobacillus–IAA–AhR signaling pathway. This study provides novel insights into colitis pathogenesis and presents an effective therapeutic and preventive approach against IBD.

Increasing plant protein in the diet induces changes in the plasma metabolome that may be beneficial for metabolic health. A randomized crossover study in males

Background & aimDietary shifts replacing animal protein (AP) with plant protein (PP) sources have been associated with lowering cardiometabolic risk (CMR), but underlying mechanisms are poorly characterized. This nutritional intervention aims to characterize the metabolic changes induced by diets containing different proportions of AP and PP sources in males at CMR. DesignThis study is a 4-week, crossover, randomized, controlled-feeding trial in which 19 males with CMR followed two diets providing either 36% for the control diet (CON-D) or 64% for the flexitarian diet (FLEX-D) of total protein intake from PP sources. Plasma nontargeted metabolomes (LC-MS method) were measured in the fasted state and after a high-fat challenge meal at the end of each intervention arm. Lipogenesis and protein synthesis fluxes, flow-mediated dilatation (FMD) and gluco-lipidic responses were assessed after the challenge meal. Data were analyzed with mixed models, and univariate and multivariate models for metabolomics data. ResultsIn both arms CMR improved with time, with decreased body weight (-0.9%), insulin resistant (-34%, HOMA-IR, Homeostatic Model Assessment for Insulin Resistance) and low-density lipoproteins (LDL)-cholesterol (-11%). Diet had no effect on FMD or metabolic fluxes, but a trend was observed for a stronger decrease in HOMA-IR and lower postprandial glucose after FLEX-D vs CON-D. The abundance of 21 and 37 metabolites differed between diets at fasted and fed states, respectively, including food intake biomarkers of AP (methylhistidine, eicosapentaenoic acid, hydroxyprolines) and PP sources (trigonelline, N-acetyl-ornithine). In fasted or fed states, indole acrylic acid and indole propionic acid, both products of tryptophan catabolism, were higher after FLEX-D vs CON-D, while the indispensable amino acids-related metabolites alpha-aminoadipic acid, hydroxymethylbutyric acids and propionylcarnitine were lower. In the postprandial state only, the ω-oxidation products dodecanedioic, tetradecanedioic and hexadecanedioic acids were higher after FLEX-D vs CON-D. ConclusionsDespite little changes in risk factors after 4 wk, this study evidenced subtle metabolic adaptations in amino acids and lipid metabolism and gut microbiota activity occurring after higher PP source intake that may be beneficial to CMR. ClinicalTrials.gov study identifierNCT04236518 Clinical Trial RegistryNCT04236518 on ClinicalTrials.gov

Intestinal barrier, immunity and gut microbiota-based protective effects of Lactococcus lactis HF08 and its postbiotic derivative on aging and aging colitis mice

The prevalence and severity of gastrointestinal diseases were increased with age. In this study, the intestinal protective effects of Lactococcus lactis HF08 (HF08) and its derived postbiotic (P-HF08) on D-gal-induced aging mice and D-gal/DSS-induced aging colitis mice were investigated. In D-gal-induced aging mice, both HF08 and P-HF08 alleviated aging-related intestinal barrier dysfunction, inflammatory status, and gut microbiota disorder. The effects of probiotic HF08 were superior to those of postbiotic P-HF08, attributed to ability of HF08 to regulate the gut microbiota. However, in D-gal/DSS-induced aging colitis mice, the effects of P-HF08 on colitis surpassed that of HF08. Specifically, both HF08 and P-HF08 could reduce symptoms of age-related colitis, including reduction of lose weight, the DAI score, colonic shortening, and colon tissue damage. The inhibitory effects of P-HF08 on intestinal inflammation surpassed those of HF08, as evidenced by the levels of colon IL-6, IL-1β, and IL-10. Western blot results demonstrated that the anti-inflammatory effects of P-HF08 were attributed to the downregulation of key proteins in the TLR4/NF-κB pathway. And four potential TLR4 inhibitors were identified from HF08 metabolites (eplerenone, genistein, indoleacrylic acid, and turanose) by molecular docking. Nevertheless, HF08 could better regulate gut microbiota and metabolite in aging-related colitis than P-HF08, which was consistent with the results on aging mice. Overall, our finding revealed that when the intestinal barrier was intact (aging), probiotics showed superior regulation of intestinal microbiota, while postbiotics offered greater safety in case of intestinal barrier damage (aging colitis). This study offered a novel perspective into the applications of probiotics and their derivatives in the aging related gastrointestinal diseases adjuvant therapy.

Integrated metabolomics and gut microbiota analysis to explore potential mechanism of Qi-Huo-Yi-Fei formula against chronic obstructive pulmonary disease

Metabolic disorders and gut microbiota dysbiosis contribute to the complicated pathology of chronic obstructive pulmonary disease (COPD). Qi-Huo-Yi-Fei formula (QHYFF) is a Chinese medicine prescription for COPD treatment and has showed beneficial clinical effects, but the underlying mechanism remains elusive. This study integrated metabolomics and gut microbiota analysis to explore potential mechanism of QHYFF against COPD. The therapeutic effects of QHYFF were evaluated using a murine model of COPD induced by cigarette smoke and lipopolysaccharide. QHYFF effectively improved pulmonary function, suppressed inflammation, and relieved lung pathological changes. Serum and urine metabolomics analysis identified 19 differential metabolites, such as L-tyrosine, epinephrine, dopamine, hypotaurine, citric acid, L-tryptophan and indoleacrylic acid, involving tyrosine metabolism, taurine and hypotaurine metabolism, citrate cycle and tryptophan metabolism. QHYFF also enriched Bifidobacterium, Blautia, Faecalibaculum and Parasutterella. Moreover, Spearman’s correlation analysis showed that discriminative metabolites and bacteria were closely correlated with efficacy indices. The findings indicated that QHYFF could be an effective therapeutic measure against COPD by regulating metabolism and gut microbiota.

Serum metabolomics analysis of malnutrition in patients with gastric cancer: a cross sectional study

BackgroundAlthough malnutrition is common in cancer patients, its molecular mechanisms has not been fully clarified. This study aims to identify significantly differential metabolites, match the corresponding metabolic pathways, and develop a predictive model of malnutrition in patients with gastric cancer.MethodsIn this cross-sectional study, we applied non-targeted metabolomics using liquid chromatography-mass spectrometry to explore the serum fingerprinting of malnutrition in patients with gastric cancer. Malnutrition-specific differential metabolites were identified by orthogonal partial least-squares discriminant analysis and t-test and matched with the Human Metabolome Database and the LIPID Metabolites and Pathways Strategy. We matched the corresponding metabolic pathways of malnutrition using pathway analysis at the MetaboAnalyst 5.0. We used random forest analyses to establish the predictive model.ResultsWe recruited 220 malnourished and 198 non-malnourished patients with gastric cancer. The intensities of 25 annotated significantly differential metabolites were lower in patients with malnutrition than those without, while two others were higher in patients with malnutrition than those without, including newly identified significantly differential metabolites such as indoleacrylic acid and lysophosphatidylcholine(18:3/0:0). We matched eight metabolic pathways associated with malnutrition, including aminoacyl-tRNA biosynthesis, tryptophan metabolism, and glycerophospholipid metabolism. We established a predictive model with an area under the curve of 0.702 (95% CI: 0.651–0.768) based on four annotated significantly differential metabolites, namely indoleacrylic acid, lysophosphatidylcholine(18:3/0:0), L-tryptophan, and lysophosphatidylcholine(20:3/0:0).ConclusionsWe identified 27 specific differential metabolites of malnutrition in malnourished compared to non-malnourished patients with gastric cancer. We also matched eight corresponding metabolic pathways and developed a predictive model. These findings provide supportive data to better understand molecular mechanisms of malnutrition in patients with gastric cancer and new strategies for the prediction, diagnosis, prevention, and treatment for those malnourished.

The MACROD2 rs6110695 A>G Polymorphism and the Metabolites Indoleacrylic Acid and Butyrylcarnitine Potentially Have Clinical Relevance to WBC Count Prediction.

Our previous study suggested that the Mono-ADP ribosylhydrolase 2 (MACROD2) rs6110695 A>G polymorphism is significantly associated with white blood cell (WBC) count in the Korean population. The present study aimed to evaluate the clinical relevance of the MACROD2 rs6110695 A>G polymorphism for predicting WBC count by utilizing plasma metabolites and a single-nucleotide polymorphism (SNP). Two groups were characterized by MACROD2 rs6110695 A>G SNP genotypes among 139 healthy subjects based on the genetic information provided in our previous work: rs6110695 AA genotype group (n = 129) and rs6110695 AG genotype group (n = 10). Plasma global metabolic profiling was performed using ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). To estimate the predictive abilities of WBC count models using the rs6110695 genotype and/or significant differential metabolites, multiple linear regression analysis and receiver operating characteristic (ROC) curve analysis were conducted. The AG genotype had greater WBC-to-apolipoprotein (apo) A-I ratios; counts of WBCs, lymphocytes, monocytes, and granulocytes; monocyte-to-lymphocyte ratio (MLR); and monocyte-to-platelet ratio (MPR) than the AA genotype. In terms of metabolic profile, indoleacetic acid, and butyrylcarnitine levels were considerably distinct between the two groups, and these metabolites were considered to be meaningful prognostic variables for the rs6110695 genotype. Finally, ROC curve analysis demonstrated that the model containing the rs6110695 genotype and the two main metabolites was reliable. The present study revealed that individuals carrying the rs6110695 AG genotype with high plasma indoleacrylic acid and butyrylcarnitine levels might have elevated WBC counts. The rs6110695 genotype and the concentrations of indoleacrylic acid and butyrylcarnitine could contribute to reducing the risk of chronic diseases in the future.

Lactobacillus murinus alleviated lung inflammation induced by PAHs in mice

ObjectiveThis study aimed to investigate the mechanism that Lactobacillus murinus (L. murinus) alleviated lung inflammation induced by polycyclic aromatic hydrocarbons (PAHs) exposure based on metabolomics. MethodsFemale mice were administrated with PAHs mix, L. murinus and indoleacrylic acid (IA) or indolealdehyde (IAId). Microbial diversity in feces was detected by 16 S rRNA gene sequencing. Non-targeted metabolomics analysis in urine samples and targeted analysis of tryptophan metabolites in serum by UPLC-Orbitrap-MS and short-chain fatty acids (SCFA) in feces by GC-MS were performed, respectively. Flow cytometry was used to determine T helper immune cell differentiation in gut and lung tissues. The levels of IgE, IL-4 and IL-17A in the bronchoalveolar lavage fluid (BALF) or serum were detected by ELISA. The expressions of aryl hydrocarbon receptor (Ahr), cytochrome P450 1A1 (Cyp1a1) and forkheadbox protein 3 (Foxp3) genes and the histone deacetylation activity were detected by qPCR and by ELISA in lung tissues, respectively. ResultsPAHs exposure induced lung inflammation and microbial composition shifts and tryptophan metabolism disturbance in mice. L. murinus alleviated PAHs-induced lung inflammation and inhibited T helper cell 17 (Th17) cell differentiation and promoted regulatory T cells (Treg) cell differentiation. L. murinus increased the levels of IA and IAId in the serum and regulated Th17/Treg imbalance by activating AhR. Additionally, L. murinus restored PAHs-induced decrease of butyric acid and valeric acid which can reduce the histone deacetylase (HDAC) level in the lung tissues, enhancing the expression of the Foxp3 gene and promoting Treg cell differentiation. Conclusionour study illustrated that L. murinus alleviated PAHs-induced lung inflammation and regulated Th17/Treg cell differentiation by regulating host tryptophan metabolism and SCFA levels. The study provided new insights into the reciprocal influence between gut microbiota, host metabolism and the immune system, suggesting that L. murinus might have the potential as a novel therapeutic strategy for lung diseases caused by environmental pollution in the future.

Metabolites of blueberry roots at different developmental stages strongly shape microbial community structure and intra-kingdom interactions at the root-soil interface

The rhizosphere microorganisms of blueberry plants have long coexisted with their hosts under distinctively acidic soil conditions, exerting a profound influence on host performance through mutualistic symbiotic interactions. Meanwhile, plants can regulate rhizosphere microorganisms by exerting host effects to meet the functional requirements of plant growth and development. However, it remains unknown how the developmental stages of blueberry plants affect the structure, function, and interactions of the rhizosphere microbial communities. Here, we examined bacterial communities and root metabolites at three developmental stages (flower and leaf bud development stage, fruit growth and development stage, and fruit maturation stage) of blueberry plants. The results revealed that the Shannon and Chao 1 indices as well as community composition varied significantly across all three developmental stages. The relative abundance of Actinobacteria significantly increased by 10 % (p < 0.05) from stage 1 to stage 2, whereas that of Proteobacteria decreased significantly. The co-occurrence network analysis revealed a relatively complex network with 1179 edges and 365 nodes in the stage 2. Niche breadth was highest at stage 2, while niche overlap tended to increase as the plant developed. Furthermore, the untargeted metabolome analysis revealed that the number of differential metabolites of vitamins, nucleic acids, steroids, and lipids increased between stage 1 to stage2 and stage 2 to stage 3, while those for differential metabolites of carbohydrates and peptides decreased. Significant changes in expression levels of levan, L-glutamic acid, indoleacrylic acid, oleoside 11-methyl ester, threo-syringoylglycerol, gingerglycolipid B, and bovinic acid were highly correlated with the bacterial community structure. Collectively, our study reveals that significant alterations in dominant bacterial taxa are strongly correlated with the dynamics of root metabolites. These findings lay the groundwork for developing prebiotic products to enhance the beneficial effects of root microorganisms and boosting blueberry productivity via a sustainable approach.

Human Milk and Dietary Supplements Differentially Modulate Microbial Communities of Stool Isolated from Term and Preterm Infants

Background: After birth, the term neonatal gut becomes colonized by commensal bacteria that benefit host health. Premature infants, however, are prone to abnormal colonization that increases the risk for neonatal intestinal disease such as necrotizing enterocolitis (NEC). Breastfeeding confers a lower risk for NEC, and this is likely due to prebiotics, probiotics, and anti-inflammatory cytokines present in breast milk. Despite the known associations of NEC with gut microbiota and breastfeeding, the exact interactions between preterm gut microbes and various milk types remains relatively unexplored. Furthermore, Lactobacillus are frequently included in probiotic supplements, but the interplay between Lactobacillus, milk, and NEC is not fully understood. Hypothesis: We hypothesize that in in vitro models of preterm gut microbiota, human breast milk exerts an overall beneficial modulatory effect on bacterial composition. We also suspect that the anti-inflammatory properties of Lactobacillus are enhanced when treated with fresh breast milk compared to formula. Methods: Stool samples from preterm infants were incubated anaerobically in mini-bioreactors overnight at 37°C to establish complex communities. The next day, they were supplemented with fresh milk with or without fortifier, donor milk with or without fortifier, formula, vitamin mix, iron mix, or water and incubated for another 24 hrs. Bacterial gDNA was examined by qPCR and back-calculated to colony-forming units (CFUs) to quantify bacterial composition. To examine the effect of milk on Lactobacillus metabolites, we monocultured six Lactobacillus ( L. acidophilus, L. breve, L. johnsonii, L. paracasei, L. reuteri, and L. rhamnosus) in a defined medium supplemented with fresh milk, formula, or water. After overnight incubation, supernatants were collected for nontargeted metabolomics. The supernatant was also applied to preterm intestinal organoids to examine epithelial responses. Results: Formula significantly elevated Bifidobacterium and Streptococcus, while fresh and donor milk caused upward trends in Lactobacillus and Bifidobacterium. LC-MS/MS data revealed that in the presence of fresh breast milk but not formula, at least one Lactobacillus strain demonstrated increased levels of anti-inflammatory metabolites such as γ-glutamylphenylalanine, ascorbic acid, vaccenic acid, hydroxyphenylacetic acid, or indoleacrylic acid. Conclusions: These data indicate that different milk types and supplements variably affect bacterial growth and metabolites in the preterm gut. This study was supported in part by the NIH K01DK123195 (MAE), NATS NIH KL2TR001452 (KEC) and UL1TR001450 (KEC) grants. This study does not represent the offcial view of the NIH. This study was also supported in part by the David and Laura Stone Endowment for Advancement in Neonatal Medicine (KEC). The Texas Children’s Hospital Department of Pathology provides salary support to the Texas Children’s Microbiome Center-Metabolomics and Proteomics Mass Spectrometry Laboratory staff, and purchased all of the reagents, the consumables and durable supplies, and the LC-MS/MS equipment described herein (TH). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Anti-adipogenesis effect of indole-3-acrylic acid on human preadipocytes and HFD-induced zebrafish.

Obesity, defined as excessive or abnormal body fat accumulation, which could significantly increase the risk of cardiovascular disease, type 2 diabetes mellitus (T2DM) diseases and seriously affect people's quality of life. More than 2 billion people are overweight, and the incidence of obesity is increasing rapidly worldwide, it has become a widely concerned public health issue in the world. Diverse evidence show that active metabolites are involved in the pathophysiological processes of obesity. However, whether the downstream catabolite of tryptophan, 3-indole acrylic acid (IA), is involved in obesity remains unclear. We collected the samples of serum from peripheral blood of obesity and health controls, and liquid chromatography-mass spectrometry (LC-MS) was performed to identify the plasma levels of IA. Additionally, we verified the potential benefits of IA on human preadipocytes and HFD- induced zebrafish by cell viability assay, flow cytometry assay, Oil red O staining, total cholesterol (T-CHO), triglyceride (TG) and nonesterified free fatty acids (NEFA) measurements and Nile Red staining. RNA-Seq, functional analysis and western blot revealed the mechanisms underlying the function of IA. We found that the content of IA in peripheral blood serum of overweight people was significantly lower than that of normal people. In addition, supplementation with IA in zebrafish larvae induced by a high fat diet (HFD) dramatically reduced HFD induced lipid accumulation. IA had no effect on proliferation and apoptosis of preadipocytes, but significantly inhibited adipogenesis of preadipocytes by down-regulate CEBPα and PPARγ. RNA-Seq and functional analysis revealed that IA regulated the adipogenesis of preadipocytes through stimulate the phosphorylation of STAT1. Taken together, IA has been identified as a potent metabolite for the prevention or treatment of obesity.

Interactions of HRLCMS-QTOF Detected Plant Metabolites of &lt;i&gt;Abrus precatorius&lt;/i&gt; L. Seeds with EGFR and the Effect of Classically Detoxification Process

Abrus precatorius L. is one of the most important medicinal plants with toxic principles which has been reported for many cases of poisoning throughout the globe. Shodhana is an important technique in Ayurveda which includes a set of procedures to detoxify the toxic herbal raw drugs. Hence three varieties of unprocessed and shodhana processed Abrus precatorius L. seeds were chosen and the hydro alcoholic extracts were prepared by maceration. These extracts were subjected to HRLCMS-QTOF analysis and the receptor-specific action of all the major components selected with the cutoff range of more than 0.2x106 counts was analyzed with GLIDE (Schrodinger 2021-1, maestro v13.6) software. Totally 570 molecules were identified in HRLCMS, and 299 molecules were docked against EGFR downloaded from the RCSB database with the crystal structure of 5XDK. The HRLCMS has revealed the presence of many unknown compounds in extracts and the major compounds identified were Indoleacrylic acid, PABA, Galangin 3- [galactosyl-(1-&gt;4), Chrysoeriol 7-O-neohesperidoside, gallic acid, calendoflaside, etc. In docking studies, the glide scores of the ligands were compared with the standard ligand 8JK and it was found that the compounds present in the extracts have shown good binding scores in comparison with the standard ligand. It is concluded that the shodhana processed seed extracts show a wide range of phytochemical variation and a notable in silico docking score and hence the raw drug after shodhana can be considered promising for research and development of cancer management therapies.

Exposure of Helicobacter pylori to clarithromycin in vitro resulting in the development of resistance and triggers metabolic reprogramming associated with virulence and pathogenicity.

In H. pylori infection, antibiotic-resistance is one of the most common causes of treatment failure. Bacterial metabolic activities, such as energy production, bacterial growth, cell wall construction, and cell-cell communication, all play important roles in antimicrobial resistance mechanisms. Identification of microbial metabolites may result in the discovery of novel antimicrobial therapeutic targets and treatments. The purpose of this work is to assess H. pylori metabolomic reprogramming in order to reveal the underlying mechanisms associated with the development of clarithromycin resistance. Previously, four H. pylori isolates were induced to become resistant to clarithromycin in vitro by incrementally increasing the concentrations of clarithromycin. Bacterial metabolites were extracted using the Bligh and Dyer technique and analyzed using metabolomic fingerprinting based on Liquid Chromatography Quadrupole Time-of-Flight Mass Spectrometry (LC-Q-ToF-MS). The data was processed and analyzed using the MassHunter Qualitative Analysis and Mass Profiler Professional software. In parental sensitivity (S), breakpoint isolates (B), and induced resistance isolates (R) H. pylori isolates, 982 metabolites were found. Furthermore, based on accurate mass, isotope ratios, abundances, and spacing, 292 metabolites matched the metabolites in the Agilent METLIN precise Mass-Personal Metabolite Database and Library (AM-PCDL). Several metabolites associated with bacterial virulence, pathogenicity, survival, and proliferation (L-leucine, Pyridoxone [Vitamine B6], D-Mannitol, Sphingolipids, Indoleacrylic acid, Dulcitol, and D-Proline) were found to be elevated in generated resistant H. pylori isolates when compared to parental sensitive isolates. The elevated metabolites could be part of antibiotics resistance mechanisms. Understanding the fundamental metabolome changes in the course of progressing from clarithromycin-sensitive to breakpoint to resistant in H. pylori clinical isolates may be a promising strategy for discovering novel alternatives therapeutic targets.

Uterine Commensal Peptostreptococcus Species Contribute to IDO1 Induction in Endometrial Cancer via Indoleacrylic Acid.

Microbial dysbiosis has an increasingly appreciated impact on carcinogenesis, and the cervicovaginal microbiome plays a critical role in microenvironmental inflammation. Here, we investigated the involvement of the female genital tract Peptostreptococcus species in gynecological cancer via indoleacrylic acid (IAA). IAA production from Peptostreptococcus species and the effect of bacterial culture on tumor growth in vivo were examined. The impact of IAA on cytokine production and indoleamine-2,3-dioxygenase 1 (IDO1) expression in an endometrial cancer (EC) cell line, as well as their effect on Treg and Teff cells, and M1 and M2 macrophage populations were examined in EC patients and tumor-grafted mice. Clinically, Peptostreptococcus species abundance, IAA, and IDO1 expression were verified in EC patients. The results showed that IAA production was induced in the uteri of BALB/c nude mice by Peptostreptococcus species transplantation, and the intratumoral injection of a conditioned medium from Peptostreptococcus cultures into tumor-grafted mice promoted tumor growth. IL-10 expression was upregulated by IAA; IFN-γ expression was increased by IL-10. IFN-γ induced IDO1 expression in the EC cell line. The co-culture of IDO1-expressing EC cells with peripheral blood mononuclear cells upregulated the Treg proportion and decreased the M1/M2 ratio. Clinically, P. anaerobius was more abundant amongst the uterine microbiota of EC patients than the control. The IAA, IDO1, and kynurenine/tryptophan ratios were all higher in EC tissue, and the M1/M2 ratio was lower. Our study sheds light on the link between IDO1 induction and uterine Peptostreptococcus dysbiosis and provides a potential rationale for the role of Peptostreptococcus species in immune tolerance induction in type I endometrial cancer.

Danshen ligustrazine injection ameliorates MCAO/R rats through the AhR-SOCS2 axis in association with gut microbiota-derived tryptophan metabolism

BackgroundDanshen Ligustrazine Injection (DSLI), composed of the extract of Salvia miltiorrhiza Bunge and Conioselinum anthriscoides 'Chuanxiong', is commonly used in the clinical treatment of ischemic stroke, which is the most common type of cerebrovascular disease, affected by multiple factors. The brain-gut axis is a new type of influencing factor for ischemic stroke, however, there is a lack of systematic research on the relationship between microbiota and host metabolic phenotype. This study aimed to investigate the host metabolic phenotype-microbiome interface in ischemic stroke and the immunomodulatory effects of DSLI. MethodsIn the study, liquid chromatography-mass spectrometry was used to primarily analyze the quality markers (Q-markers) and the polyphenols of DSLI. Based on 16S rRNA gene sequencing and metabolomics, the variations of microbial composition in different groups were characterized. In addition, a network diagram was constructed to reveal the interrelationship between DSLI-regulated gut microbiota changes and DSLI-regulated brain metabolites in MCAO/R rats. The expression of the aryl hydrocarbon receptor (AhR) in the colon and brain by immunofluorescence and suppressors of cytokine signaling 2 (SOCS2) by western blot were further detected. ResultsDiminished metabolic diversity and alterations in the tryptophan metabolism pathway were a feature of IS in the study. The results revealed that Clostri_UCG-014 was positively correlated with indoleacrylic acid, indole-3-methylacetate, and Ruminococcaceae was strongly related with L-tryptophan, 3-methylindole, and indole. The results of immunofluorescence showed that the expression of AhR in the colon and brain tissues of rats in the model group was greatly reduced compared with that in the sham-operated group, and was heavily activated with the treatment of DSLI. The levels of aryl hydrocarbon nuclear receptor ARNT2 and SOCS2 were down-regulated in the brains of Model rats, whereas increased by DSLI, further confirming that the brain-gut microbiota-Trp metabolism exists in the pathological changes of stroke and the intervention mechanism of DSLI. ConclusionsThe present study findings suggest Clostri_UCG-014, Ruminococcaceae, and Firmicute_RF39 play an important role in improving the regulation of immune function in stroke through tryptophan metabolites by acting on the AhR-SOCS2 axis.

Metabolic changes preceding bladder cancer occurrence among Korean men: a nested case-control study from the KCPS-II cohort

BackgroundBladder cancer (BLCA) research in Koreans is still lacking, especially in focusing on the prediction of BLCA. The current study aimed to discover metabolic signatures related to BLCA onset and confirm its potential as a biomarker.MethodsWe designed two nested case-control studies using Korean Cancer Prevention Study (KCPS)-II. Only males aged 35–69 were randomly selected and divided into two sets by recruitment organizations [set 1, BLCA (n = 35) vs. control (n = 35); set 2, BLCA (n = 31) vs. control (n = 31)]. Baseline serum samples were analyzed by non-targeted metabolomics profiling, and OPLS-DA and network analysis were performed. Calculated genetic risk score (GRS) for BLCA from all KCPS participants was utilized for interpreting metabolomics data.ResultsCritical metabolic signatures shown in the BLCA group were dysregulation of lysine metabolism and tryptophan-indole metabolism. Furthermore, the prediction model consisting of metabolites (lysine, tryptophan, indole, indoleacrylic acid, and indoleacetaldehyde) reflecting these metabolic signatures showed mighty BLCA predictive power (AUC: 0.959 [0.929–0.989]). The results of metabolic differences between GRS-high and GRS-low groups in BLCA indicated that the pathogenesis of BLCA is associated with a genetic predisposition. Besides, the predictive ability for BLCA on the model using GRS and five significant metabolites was powerful (AUC: 0.990 [0.980–1.000]).ConclusionMetabolic signatures shown in the present research may be closely associated with BLCA pathogenesis. Metabolites involved in these could be predictive biomarkers for BLCA. It could be utilized for early diagnosis, prognostic diagnosis, and therapeutic targets for BLCA.

Thyme Extract Alleviates High-Fat Diet-Induced Obesity and Gut Dysfunction

Prolonged intake of a high-fat diet (HFD) disturbs the composition of gut microbiota, contributing to the development of metabolic diseases, notably obesity and increased intestinal permeability. Thyme (Thymus vulgaris L.), an aromatic plant, is known for its several therapeutic properties. In this study, we explored the potential of thyme extract (TLE) to mitigate HFD-induced metabolic derangements and improve the gut environment. Eight-week-old C57BL/6 mice were administered 50 or 100 mg/kg TLE for eight weeks. Administration of 100 mg/kg TLE resulted in decreased weight gain and body fat percentage, alongside the regulation of serum biomarkers linked to obesity induced by a HFD. Moreover, TLE enhanced intestinal barrier function by increasing the expression of tight junction proteins and ameliorated colon shortening. TLE also altered the levels of various metabolites. Especially, when compared with a HFD, it was confirmed that 2-hydroxypalmitic acid and 3-indoleacrylic acid returned to normal levels after TLE treatment. Additionally, we investigated the correlation between fecal metabolites and metabolic parameters; deoxycholic acid displayed a positive correlation with most parameters, except for colon length. In contrast, hypoxanthine was negatively correlated with most parameters. These results suggest a promising role for thyme in ameliorating obesity and related gut conditions associated with a HFD.

Changes in urine metabolite concentration as a minimally invasive marker of ovarian serous adenocarcinoma

Introduction. Detection of ovarian cancer (OC) at the earliest possible stages is a priority for gynecological oncology, since 5-year survival rates decrease significantly with the progression of the disease. Currently, there is a huge need for more effective diagnostic methods and approaches. In recent years, fluid biopsy has received increasing attention in precision medicine because it is minimally invasive and can be repeated many times, allowing for realtime disease monitoring.Aim. Study of the urine metabolomic profile of patients with ovarian carcinoma.Materials and methods. To perform metabolomic analysis, 50 urine samples from patients with a diagnosis of serous ovarian carcinoma and 20 samples from apparently healthy individuals were selected. For protein precipitation, 300 mkl of urine was mixed with 600 mkl of a solution of acetonitrile LC-MS (Merck, Germany) and methanol LC-MS (Merck, Germany) (3:1 ratio). Chromatographic separation of metabolites was performed on a Vanquish Flex UHPLC System chromatograph (Thermo Scientific, Germany). The chromatograph was coupled to an Orbitrap Exploris 480 mass spectrometer (Thermo Scientific, Germany) equipped with an electrospray ionization source. Chromatographic separation was carried out on a Hypersil GOLD™ C18 column (1.9mkm, 10 x 2.1 mm) using the following eluents: A, 0.1 % formic acid; B, acetonitrile containing 0.1 % formic acid.Results. A total of 417 metabolites of various classes were identified by HPLC-MS. It was shown that in the urine of patients with OC 14 metabolites (kynurenine, phenylalanyl-valine, lysophosphatidylcholine (18:3), lysophosphatidylcholine (18:2), alanyl-leucine, lysophosphatidylcholine (20:4), L-phenylalanine, phosphatidylinositol (34:1), 5-methoxytryptophan, 2-hydroxymyristic acid, 3-oxocholic acid, lysophosphatidylcholine (14:0), indoleacrylic acid, lysophosphatidylserine (20:4)) had a significantly higher concentration compared to apparently healthy individuals. The content of 12 compounds, on the contrary, was reduced (L-beta-aspartyl-L-phenylalanine, myristic acid, decanoylcarnitine, aspartyl-glycine, malonylcarnitine, 3-hydroxybutyrylcarnitine, 3-methylxanthine, 2,6-dimethylheptanoylcarnitine, 3-oxododecanoic acid, N-acetylproline, L-octanoylcarnitine, capryloylglycine). This indicates a significant metabolomic imbalance in patients with OC.Conclusion. The metabolomic profile study of urine by UHPLC-MS showed that in patients with serous ovarian carcinoma there is an imbalance in the content of certain fatty acids and their derivatives, acylcarnitines, phospholipids, amino acids and their derivatives, as well as some derivatives of nitrogenous bases. At the same time, 26 metabolites with abnormal concentrations in urine may have some potential as non-invasive biomarkers of OC in women belonging to high-risk groups.

A proof-of-concept study on the versatility of liquid chromatography coupled to high-resolution mass spectrometry to screen for various contaminants and highlight markers of floral and geographical origin for different honeys

The high-resolution mass spectrometry is a powerful analytical tool for improving food safety and authenticity, but still underused in official control laboratories. The present work is a proof-of-concept study overviewing how liquid-chromatography coupled to high-resolution mass spectrometry could be used simultaneously for large-scale screening of contaminants and differentiation of honey samples. Within this study, the samples were extracted using all-in-one QuEChERS-based protocol that allowed for analysis of various anthropogenic contaminants and endogenous compounds. First, targeted-analysis of 52 honey samples led to unequivocal identification of 23 chemicals, including neonicotinoids, triazole fungicides and synergist. Then, suspect-screening using MSDial software allowed for tentative identification of 30 chemicals including plasticizers, flame-retardants and additives. Suspect-screening also made it possible to highlight tentative markers of chestnut honey (deoxyvasicinone, 2-quinolone, indoleacrylic acid and kynurenic acid) and citrus honey (caffeine, 2-oxindole and indole-3-carbinol). Lastly, non-targeted analysis enabled to separate honeys by their type, floral and geographical origins.

Integrated physiological and metabolomic analyses reveal changes during the natural senescence of Quercus mongolica leaves.

Quercus mongolica is a common landscape, afforestation, and construction timber species in northern China with high ecological, economic, and ornamental value. Leaf senescence is a complex process that has important implications for plant growth and development. To explore changes of metabolites during the ageing of Quercus mongolica leaves, we investigated physiological responses and metabolite composition in ageing leaves harvested from 15-20-year-old Quercus mongolica. Leaf samples of Q. mongolica were collected when they were still green (at maturity) (stage 1), during early senescence (stage 2), and during late senescence (stage 3). These leaves were then subjected to physiological index and metabolome sequencing analyses. The physiological analysis showed that the leaves of Q. mongolica changed from green to yellow during senescence, which induced significant accumulation of soluble sugar and significant reductions in the concentration of soluble protein and chlorophyll. Peroxidase and catalase were the main antioxidant enzymes mitigating leaf senescence. Metabolomic analysis identified 797 metabolites during leaf senescence. Compared to stage 1, 70 differential metabolites were screened in stage 2 and 72 were screened in stage 3. Differential metabolites in the two senescent stages were principally enriched in amino acid metabolism, lipid metabolism and secondary metabolite biosynthesis. The contents of N-oleoylethanolamine and N, N-dimethylglycine were significantly increased only in stage 2, while the contents of trifolin, astragalin, valine, isoleucine, leucine, and citric acid were significantly increased only in stage 3. Histidine, homoserine, tryptophan, tyrosine, phenylalanine, proline, norleucine, N-glycyl-L-leucine, linoleic acid, linolenic acid, gallic acid, 3-indoleacrylic acid, 3-amino-2-naphthoic acid, 3-hydroxy-3-methylpentane-1,5-dioic acid, 2,3,4-trihydroxybenzoic acid, trifolin, astragalin, DL-2-aminoadipic acid, pinoresinol dimethyl ether, dimethylmatairesinol, and lysophosphatidylcholine increased during both stage 2 and stage 3. Increasing contents of these metabolites may constitute the main mechanism by which Q. mongolica leaves adapt to senescence.