Integrative Molecular Phenotyping
INTEGRATIVE MOLECULAR
PHENOTYPING
WHEELOCK LABORATORY
DEPARTMENT OF MEDICAL
BIOCHEMISTRY AND BIOPHYSICS
WHEELOCK LABORATORY
DEPARTMENT OF MEDICAL
BIOCHEMISTRY AND BIOPHYSICS
WHEELOCK LABORATORY
DEPARTMENT OF MEDICAL
BIOCHEMISTRY AND BIOPHYSICS
WHEELOCK LABORATORY
DEPARTMENT OF MEDICAL
BIOCHEMISTRY AND BIOPHYSICS
WHEELOCK LABORATORY
DEPARTMENT OF MEDICAL
BIOCHEMISTRY AND BIOPHYSICS
WHEELOCK LABORATORY

PubMed

Use of Multivariate Analysis to Unravel the Differences between Two Chamomile Varieties and Their Anticancer and Antioxidant Activities

Wed, 28/06/2023 - 12:00
Plants (Basel). 2023 Jun 12;12(12):2297. doi: 10.3390/plants12122297.ABSTRACTBACKGROUND: Plants from the Asteraceae family were commonly used to treat various diseases. The metabolomic profile of this family consisted of bioactive flavonoids and other phenolics. Chamomile is a member of the Asteraceae family. Jordanian and European chamomile are two varieties of Matricaria chamomilla (German chamomile), which were grown under different environmental conditions, were studied. Many examples of plant varieties with significant distinction in the secondary metabolite they afford have been described in the literature. Multivariate statistical analysis was employed to measure the depth of this variation in two chamomile varieties.METHODS: From both types, crude extracts were prepared using solvents of different polarities and tested for their biological activity. The semipolar fraction of the European variety showed anticancer and antioxidant activity. Meanwhile, the semipolar fraction of the Jordanian type exhibited only antioxidant activity. Both extracts were fractionated, and then the biological activity was again assayed.RESULTS: European and Jordanian chamomile fractions produced dicaffeoylquinic acid isomers exhibiting antioxidant capability. Additionally, Z-glucoferulic acid was produced from the European chamomile, demonstrating antioxidant activity. The European samples afforded two major compounds, chrysosplenetin and apigenin, that displayed anticancer activity.CONCLUSIONS: Different environmental conditions between Jordanian and European chamomile affected the type of isolated compounds. Structure elucidation was performed with HPLC-MS coupled with dereplication techniques and 2D NMR experiments.PMID:37375922 | DOI:10.3390/plants12122297

The Impact of Growth Years on the Medicinal Material Characteristics and Metabolites of <em>Stellaria dichotoma</em> L. var. <em>lanceolata</em> Bge. Reveals the Optimal Harvest Age

Wed, 28/06/2023 - 12:00
Plants (Basel). 2023 Jun 12;12(12):2286. doi: 10.3390/plants12122286.ABSTRACTThe original plant of Chinese medicine Stellariae Radix (Yin Chai Hu) is Stellaria dichotoma L. var. lanceolata Bge (abbreviated as SDL). SDL is a perennial herbaceous plant and a characteristic crop in Ningxia. Growth years are vital factors that affect the quality of perennial medicinal materials. This study aims to investigate the impact of growth years on SDL and screen for the optimal harvest age by comparing the medicinal material characteristics of SDL with different growth years. Additionally, metabolomics analysis using UHPLC-Q-TOF MS was employed to investigate the impact of growth years on the accumulation of metabolites in SDL. The results show that the characteristics of medicinal materials and the drying rate of SDL gradually increase with the increase in growth years. The fastest development period of SDL occurred during the first 3 years, after which the development slowed down. Medicinal materials characteristics of 3-year-old SDL exhibited mature qualities with a high drying rate, methanol extract content, and the highest content of total sterols and total flavonoids. A total of 1586 metabolites were identified, which were classified into 13 major classes with more than 50 sub-classes. Multivariate statistical analysis indicated significant differences in the diversity of metabolites of SDL in different growth years, with greater differences observed in metabolites as the growth years increased. Moreover, different highly expressed metabolites in SDL at different growth years were observed: 1-2 years old was beneficial to the accumulation of more lipids, while 3-5 years old was conducive to accumulating more alkaloids, benzenoids, etc. Furthermore, 12 metabolites accumulating with growth years and 20 metabolites decreasing with growth years were screened, and 17 significantly different metabolites were noted in 3-year-old SDL. In conclusion, growth years not only influenced medicinal material characteristics, drying rate, content of methanol extract, and total sterol and flavonoid contents, but also had a considerable effect on SDL metabolites and metabolic pathways. SDL planted for 3 years presented the optimum harvest time. The screened significantly different metabolites with biological activity, such as rutin, cucurbitacin e, isorhamnetin-3-o-glucoside, etc., can be utilized as potential quality markers of SDL. This research provides references for studying the growth and development of SDL medicinal materials, the accumulation of metabolites, and the selection of optimal harvest time.PMID:37375910 | DOI:10.3390/plants12122286

Chlorogenic Acid Induced Neuroblastoma Cells Differentiation via the ACAT1-TPK1-PDH Pathway

Wed, 28/06/2023 - 12:00
Pharmaceuticals (Basel). 2023 Jun 14;16(6):877. doi: 10.3390/ph16060877.ABSTRACTBACKGROUND: Chlorogenic acid (CHA) has been shown to have substantial biological activities, including anti-inflammatory, antioxidant, and antitumor effects. However, the pharmacological role of CHA in neuroblastoma has not yet been assessed. Neuroblastoma is a type of cancer that develops in undifferentiated sympathetic ganglion cells. This study aims to assess the antitumor activity of CHA against neuroblastoma and reveal its mechanism of action in cell differentiation.METHODS: Be(2)-M17 and SH-SY5Y neuroblastoma cells were used to confirm the differentiation phenotype. Subcutaneous and orthotopic xenograft mouse models were also used to evaluate the antitumor activity of CHA. Seahorse assays and metabolomic analyses were further performed to investigate the roles of CHA and its target ACAT1 in mitochondrial metabolism.RESULTS: CHA induced the differentiation of Be(2)-M17 and SH-SY5Y neuroblastoma cells in vivo and in vitro. The knockdown of mitochondrial ACAT1, which was inhibited by CHA, also resulted in differentiation characteristics in vivo and in vitro. A metabolomic analysis revealed that thiamine metabolism was involved in the differentiation of neuroblastoma cells.CONCLUSIONS: These results provide evidence that CHA shows good antitumor activity against neuroblastoma via the induction of differentiation, by which the ACAT1-TPK1-PDH pathway is involved. CHA is a potential drug candidate for neuroblastoma therapy.PMID:37375824 | DOI:10.3390/ph16060877

Study on the Intervention Mechanism of Cryptotanshinone on Human A2780 Ovarian Cancer Cell Line Using GC-MS-Based Cellular Metabolomics

Wed, 28/06/2023 - 12:00
Pharmaceuticals (Basel). 2023 Jun 9;16(6):861. doi: 10.3390/ph16060861.ABSTRACTCryptotanshinone (CT), an active component of the traditional Chinese medicine Salvia miltiorrhiza Bunge, exhibits a wide range of biological and pharmacological activities. Although the anticancer activity of CT is well known, the knowledge of its effect on the regulation of cancer cell metabolism is relatively new. The present study investigated the anticancer mechanism of CT in ovarian cancer with a focus on cancer metabolism. CCK8 assays, apoptosis assays, and cell cycle assays were conducted to reveal the growth-suppressive effect of CT on ovarian cancer A2780 cells. To explore the potential underlying mechanisms of CT, the changes in endogenous metabolites in A2780 cells before and after CT intervention were investigated using the gas chromatography-mass spectrometry (GC-MS) approach. A total of 28 important potential biomarkers underwent significant changes, mainly involving aminoacyl-tRNA biosynthesis, energy metabolism, and other pathways. Changes in the ATP and amino acid contents were verified with in vitro and in vivo experiments. Our results indicate that CT may exert an anti-ovarian cancer effect by inhibiting ATP production, promoting the protein catabolic process, and inhibiting protein synthesis, which may lead to cell cycle arrest and apoptosis.PMID:37375808 | DOI:10.3390/ph16060861

Metabolomics and Network Analyses Reveal Phenylalanine and Tyrosine as Signatures of Anthracycline-Induced Hepatotoxicity

Wed, 28/06/2023 - 12:00
Pharmaceuticals (Basel). 2023 May 26;16(6):797. doi: 10.3390/ph16060797.ABSTRACTThe chemotherapy drug doxorubicin (DOX) is an anthracycline with over 30% incidence of liver injury in breast cancer patients, yet the mechanism of its hepatotoxicity remains unclear. To identify potential biomarkers for anthracycline-induced hepatotoxicity (AIH), we generated clinically-relevant mouse and rat models administered low-dose, long-term DOX. These models exhibited significant liver damage but no decline in cardiac function. Through untargeted metabolic profiling of the liver, we identified 27 differential metabolites in a mouse model and 28 in a rat model. We then constructed a metabolite-metabolite network for each animal model and computationally identified several potential metabolic markers, with particular emphasis on aromatic amino acids, including phenylalanine, tyrosine, and tryptophan. We further performed targeted metabolomics analysis on DOX-treated 4T1 breast cancer mice for external validation. We found significant (p < 0.001) reductions in hepatic levels of phenylalanine and tyrosine (but not tryptophan) following DOX treatment, which were strongly correlated with serum aminotransferases (ALT and AST) levels. In summary, the results of our study present compelling evidence supporting the use of phenylalanine and tyrosine as metabolic signatures of AIH.PMID:37375744 | DOI:10.3390/ph16060797

Dietary Docosahexaenoic Acid and Glucose Systemic Metabolic Changes in the Mouse

Wed, 28/06/2023 - 12:00
Nutrients. 2023 Jun 8;15(12):2679. doi: 10.3390/nu15122679.ABSTRACTThe endocannabinoid system (ECS) participates in regulating whole body energy balance. Overactivation of the ECS has been associated with the negative consequence of obesity and type 2 diabetes. Since activators of the ECS rely on lipid-derived ligands, an investigation was conducted to determine whether dietary PUFA could influence the ECS to affect glucose clearance by measuring metabolites of macronutrient metabolism. C57/blk6 mice were fed a control or DHA-enriched semi-purified diet for a period of 112 d. Plasma, skeletal muscle, and liver were collected after 56 d and 112 d of feeding the diets for metabolomics analysis. Key findings characterized a shift in glucose metabolism and greater catabolism of fatty acids in mice fed the DHA diet. Glucose use and promotion of fatty acids as substrate were found based on levels of metabolic pathway intermediates and altered metabolic changes related to pathway flux with DHA feeding. Greater levels of DHA-derived glycerol lipids were found subsequently leading to the decrease of arachidonate-derived endocannabinoids (eCB). Levels of 1- and 2-arachidonylglcerol eCB in muscle and liver were lower in the DHA diet group compared to controls. These findings demonstrate that DHA feeding in mice alters macronutrient metabolism and may restore ECS tone by lowering arachidonic acid derived eCB.PMID:37375583 | DOI:10.3390/nu15122679

Integrated Transcriptomic and Metabolomic Analyses Reveal the Effects of Grafting on Special Metabolites of <em>Acanthopanax senticosus</em> Leaves

Wed, 28/06/2023 - 12:00
Molecules. 2023 Jun 20;28(12):4877. doi: 10.3390/molecules28124877.ABSTRACTAcanthopanax senticosus (A. senticosus) is a member of Acanthopanax Miq. and is used in traditional Chinese medicine, and it has been found that grafting technology can be used to alter plant metabolite composition and transcriptome characteristics. In this study, shoots of A. senticosus were grafted onto the rootstocks of the vigorous Acanthopanax sessiliflorus (A. sessiliflorus) to improve its varietal characteristics. In order to investigate the changes in metabolites and transcriptional patterns in grafted A. senticosus leaves (GSCL), fresh leaves were collected from 2-year-old grafted A. senticosus scions, while self-rooted seedling A. senticosus leaves (SCL) were used as controls to analyse the transcriptome and metabolome. Metabolic profiles and gene expression patterns were further identified and correlated in special metabolite target pathways. The content of chlorogenic acid and triterpenoids in the GSCL was higher than in the control, while the quercetin content was lower. All these metabolic changes were associated with changes in the expression pattern of transcripts. Our results revealed the transcriptome and metabolome characteristics of GSCL. This may help to improve leaf quality in A. senticosus cultivation, suggesting that it is feasible to improve the medicinal quality of GSCL through asexual propagation, but the long-term effects need further investigation. In conclusion, this dataset provides a useful resource for future studies on the effects of grafting on medicinal plants.PMID:37375432 | DOI:10.3390/molecules28124877

Artefact Profiling: Panomics Approaches for Understanding the Materiality of Written Artefacts

Wed, 28/06/2023 - 12:00
Molecules. 2023 Jun 20;28(12):4872. doi: 10.3390/molecules28124872.ABSTRACTThis review explains the strategies behind genomics, proteomics, metabolomics, metallomics and isotopolomics approaches and their applicability to written artefacts. The respective sub-chapters give an insight into the analytical procedure and the conclusions drawn from such analyses. A distinction is made between information that can be obtained from the materials used in the respective manuscript and meta-information that cannot be obtained from the manuscript itself, but from residues of organisms such as bacteria or the authors and readers. In addition, various sampling techniques are discussed in particular, which pose a special challenge in manuscripts. The focus is on high-resolution, non-targeted strategies that can be used to extract the maximum amount of information about ancient objects. The combination of the various omics disciplines (panomics) especially offers potential added value in terms of the best possible interpretations of the data received. The information obtained can be used to understand the production of ancient artefacts, to gain impressions of former living conditions, to prove their authenticity, to assess whether there is a toxic hazard in handling the manuscripts, and to be able to determine appropriate measures for their conservation and restoration.PMID:37375427 | DOI:10.3390/molecules28124872

NMR-Based Metabolomic Study on <em>Phaseolus vulgaris</em> Flour Fermented by Lactic Acid Bacteria and Yeasts

Wed, 28/06/2023 - 12:00
Molecules. 2023 Jun 20;28(12):4864. doi: 10.3390/molecules28124864.ABSTRACTIn recent years, fermented foods have attracted increasing attention due to their important role in the human diet, since they supply beneficial health effects, providing important sources of nutrients. In this respect, a comprehensive characterization of the metabolite content in fermented foods is required to achieve a complete vision of physiological, microbiological, and functional traits. In the present preliminary study, the NMR-based metabolomic approach combined with chemometrics has been applied, for the first time, to investigate the metabolite content of Phaseolus vulgaris flour fermented by different lactic acid bacteria (LAB) and yeasts. A differentiation of microorganisms (LAB and yeasts), LAB metabolism (homo- and heterofermentative hexose fermentation), LAB genus (Lactobacillus, Leuconostoc, and Pediococcus), and novel genera (Lacticaseibacillus, Lactiplantibacillus, and Lentilactobacillus) was achieved. Moreover, our findings showed an increase of free amino acids and bioactive molecules, such as GABA, and a degradation of anti-nutritional compounds, such as raffinose and stachyose, confirming the beneficial effects of fermentation processes and the potential use of fermented flours in the production of healthy baking foods. Finally, among all microorganisms considered, the Lactiplantibacillus plantarum species was found to be the most effective in fermenting bean flour, as a larger amount of free amino acids were assessed in their analysis, denoting more intensive proteolytic activity.PMID:37375419 | DOI:10.3390/molecules28124864

Exploring Proton-Only NMR Experiments and Filters for Daphnia In Vivo: Potential and Limitations

Wed, 28/06/2023 - 12:00
Molecules. 2023 Jun 20;28(12):4863. doi: 10.3390/molecules28124863.ABSTRACTEnvironmental metabolomics provides insight into how anthropogenic activities have an impact on the health of an organism at the molecular level. Within this field, in vivo NMR stands out as a powerful tool for monitoring real-time changes in an organism's metabolome. Typically, these studies use 2D 13C-1H experiments on 13C-enriched organisms. Daphnia are the most studied species, given their widespread use in toxicity testing. However, with COVID-19 and other geopolitical factors, the cost of isotope enrichment increased ~6-7 fold over the last two years, making 13C-enriched cultures difficult to maintain. Thus, it is essential to revisit proton-only in vivo NMR and ask, "Can any metabolic information be obtained from Daphnia using proton-only experiments?". Two samples are considered here: living and whole reswollen organisms. A range of filters are tested, including relaxation, lipid suppression, multiple-quantum, J-coupling suppression, 2D 1H-1H experiments, selective experiments, and those exploiting intermolecular single-quantum coherence. While most filters improve the ex vivo spectra, only the most complex filters succeed in vivo. If non-enriched organisms must be used, then, DREAMTIME is recommended for targeted monitoring, while IP-iSQC was the only experiment that allowed non-targeted metabolite identification in vivo. This paper is critically important as it documents not just the experiments that succeed in vivo but also those that fail and demonstrates first-hand the difficulties associated with proton-only in vivo NMR.PMID:37375418 | DOI:10.3390/molecules28124863

Omics-Based Investigations of Breast Cancer

Wed, 28/06/2023 - 12:00
Molecules. 2023 Jun 14;28(12):4768. doi: 10.3390/molecules28124768.ABSTRACTBreast cancer (BC) is characterized by an extensive genotypic and phenotypic heterogeneity. In-depth investigations into the molecular bases of BC phenotypes, carcinogenesis, progression, and metastasis are necessary for accurate diagnoses, prognoses, and therapy assessments in predictive, precision, and personalized oncology. This review discusses both classic as well as several novel omics fields that are involved or should be used in modern BC investigations, which may be integrated as a holistic term, onco-breastomics. Rapid and recent advances in molecular profiling strategies and analytical techniques based on high-throughput sequencing and mass spectrometry (MS) development have generated large-scale multi-omics datasets, mainly emerging from the three "big omics", based on the central dogma of molecular biology: genomics, transcriptomics, and proteomics. Metabolomics-based approaches also reflect the dynamic response of BC cells to genetic modifications. Interactomics promotes a holistic view in BC research by constructing and characterizing protein-protein interaction (PPI) networks that provide a novel hypothesis for the pathophysiological processes involved in BC progression and subtyping. The emergence of new omics- and epiomics-based multidimensional approaches provide opportunities to gain insights into BC heterogeneity and its underlying mechanisms. The three main epiomics fields (epigenomics, epitranscriptomics, and epiproteomics) are focused on the epigenetic DNA changes, RNAs modifications, and posttranslational modifications (PTMs) affecting protein functions for an in-depth understanding of cancer cell proliferation, migration, and invasion. Novel omics fields, such as epichaperomics or epimetabolomics, could investigate the modifications in the interactome induced by stressors and provide PPI changes, as well as in metabolites, as drivers of BC-causing phenotypes. Over the last years, several proteomics-derived omics, such as matrisomics, exosomics, secretomics, kinomics, phosphoproteomics, or immunomics, provided valuable data for a deep understanding of dysregulated pathways in BC cells and their tumor microenvironment (TME) or tumor immune microenvironment (TIMW). Most of these omics datasets are still assessed individually using distinct approches and do not generate the desired and expected global-integrative knowledge with applications in clinical diagnostics. However, several hyphenated omics approaches, such as proteo-genomics, proteo-transcriptomics, and phosphoproteomics-exosomics are useful for the identification of putative BC biomarkers and therapeutic targets. To develop non-invasive diagnostic tests and to discover new biomarkers for BC, classic and novel omics-based strategies allow for significant advances in blood/plasma-based omics. Salivaomics, urinomics, and milkomics appear as integrative omics that may develop a high potential for early and non-invasive diagnoses in BC. Thus, the analysis of the tumor circulome is considered a novel frontier in liquid biopsy. Omics-based investigations have applications in BC modeling, as well as accurate BC classification and subtype characterization. The future in omics-based investigations of BC may be also focused on multi-omics single-cell analyses.PMID:37375323 | DOI:10.3390/molecules28124768

LC-MS Metabolomic Profiling of Five Types of Unrefined, Cold-Pressed Seed Oils to Identify Markers to Determine Oil Authenticity and to Test for Oil Adulteration

Wed, 28/06/2023 - 12:00
Molecules. 2023 Jun 14;28(12):4754. doi: 10.3390/molecules28124754.ABSTRACTThe authenticity of food products marketed as health-promoting foods-especially unrefined, cold-pressed seed oils-should be controlled to ensure their quality and safeguard consumers and patients. Metabolomic profiling using liquid chromatography coupled to quadrupole time-of-flight mass spectrometry (LC-QTOF) was employed to identify authenticity markers for five types of unrefined, cold-pressed seed oils: black seed oil (Nigella sativa L.), pumpkin seed oil (Cucurbita pepo L.), evening primrose oil (Oenothera biennis L.), hemp oil (Cannabis sativa L.) and milk thistle oil (Silybum marianum). Of the 36 oil-specific markers detected, 10 were established for black seed oil, 8 for evening primrose seed oil, 7 for hemp seed oil, 4 for milk thistle seed oil and 7 for pumpkin seed oil. In addition, the influence of matrix variability on the oil-specific metabolic markers was examined by studying binary oil mixtures containing varying volume percentages of each tested oil and each of three potential adulterants: sunflower, rapeseed and sesame oil. The presence of oil-specific markers was confirmed in 7 commercial oil mix products. The identified 36 oil-specific metabolic markers proved useful for confirming the authenticity of the five target seed oils. The ability to detect adulterations of these oils with sunflower, rapeseed and sesame oil was demonstrated.PMID:37375308 | DOI:10.3390/molecules28124754

Effect of Frost on the Different Metabolites of Two Mulberry (<em>Morus nigra</em> L. and <em>Morus alba</em> L.) Leaves

Wed, 28/06/2023 - 12:00
Molecules. 2023 Jun 12;28(12):4718. doi: 10.3390/molecules28124718.ABSTRACTMulberry leaves are a well-known traditional Chinese medicine herb, and it has been observed since ancient times that leaves collected after frost have superior medicinal properties. Therefore, understanding the changes in critical metabolic components of mulberry leaves, specifically Morus nigra L., is essential. In this study, we conducted widely targeted metabolic profiling analyses on two types of mulberry leaves, including Morus nigra L. and Morus alba L., harvested at different times. In total, we detected over 100 compounds. After frost, 51 and 58 significantly different metabolites were identified in the leaves of Morus nigra L. and Morus alba L., respectively. Further analysis revealed a significant difference in the effect of defrosting on the accumulation of metabolites in the two mulberries. Specifically, in Morus nigra L., the content of 1-deoxynojirimycin (1-DNJ) in leaves decreased after frost, while flavonoids peaked after the second frost. In Morus alba L., the content of DNJ increased after frost, reaching its peak one day after the second frost, whereas flavonoids primarily peaked one week before frost. In addition, an analysis of the influence of picking time on metabolite accumulation in two types of mulberry leaves demonstrated that leaves collected in the morning contained higher levels of DNJ alkaloids and flavonoids. These findings provide scientific guidance for determining the optimal harvesting time for mulberry leaves.PMID:37375273 | DOI:10.3390/molecules28124718

Post-Ripening and Key Glycosyltransferase Catalysis to Promote Sweet Mogrosides Accumulation of <em>Siraitia grosvenorii</em> Fruits

Wed, 28/06/2023 - 12:00
Molecules. 2023 Jun 11;28(12):4697. doi: 10.3390/molecules28124697.ABSTRACTSweet mogrosides are not only the primary bioactive ingredient in Siraitia grosvenorii fruits that exhibit anti-tussive properties and expectorate phlegm, but they are also responsible for the fruit's sweetness. Increasing the content or proportion of sweet mogrosides in Siraitia grosvenorii fruits is significant for improving their quality and industrial production. Post-ripening is an essential step in the post-harvest processing of Siraitia grosvenorii fruits, but the underlying mechanism and condition of post-ripening on Siraitia grosvenorii quality improvement need to be studied systematically. Therefore, this study analyzed the mogroside metabolism in Siraitia grosvenorii fruits under different post-ripening conditions. We further examined the catalytic activity of glycosyltransferase UGT94-289-3 in vitro. The results showed that the post-ripening process of fruits could catalyze the glycosylation of bitter-tasting mogroside IIE and III to form sweet mogrosides containing four to six glucose units. After ripening at 35 °C for two weeks, the content of mogroside V changed significantly, with a maximum increase of 80%, while the increase in mogroside VI was over twice its initial amount. Furthermore, under the suitable catalytic condition, UGT94-289-3 could efficiently convert the mogrosides with less than three glucose units into structurally diverse sweet mogrosides, i.e., with mogroside III as the substrate, 95% of it can converted into sweet mogrosides. These findings suggest that controlling the temperature and related catalytic conditions may activate UGT94-289-3 and promote the accumulation of sweet mogrosides. This study provides an effective method for improving the quality of Siraitia grosvenorii fruits and the accumulation of sweet mogrosides, as well as a new economical, green, and efficient method for producing sweet mogrosides.PMID:37375251 | DOI:10.3390/molecules28124697

Metabolic Fingerprinting of Muscat of Alexandria Grape Musts during Industrial Alcoholic Fermentation Using HS-SPME and Liquid Injection with TMS Derivatization GC-MS Methods

Wed, 28/06/2023 - 12:00
Molecules. 2023 Jun 8;28(12):4653. doi: 10.3390/molecules28124653.ABSTRACTMuscat of Alexandria is one of the most aromatic grape cultivars, with a characteristic floral and fruity aroma, producing popular appellation of origin wines. The winemaking process is a critical factor contributing to the quality of the final product, so the aim of this work was to study metabolomic changes during the fermentation of grape musts at the industrial level from 11 tanks, 2 vintages, and 3 wineries of Limnos Island. A Headspace Solid-Phase Microextraction (HS-SPME) and a liquid injection with Trimethylsilyl (TMS) derivatization Gas Chromatography-Mass Spectrometry (GC-MS) methods were applied for the profiling of the main volatile and non-volatile polar metabolites originating from grapes or produced during winemaking, resulting in the identification of 109 and 69 metabolites, respectively. Multivariate statistical analysis models revealed the differentiation between the four examined time points during fermentation, and the most statistically significant metabolites were investigated by biomarker assessment, while their trends were presented with boxplots. Whilst the majority of compounds (ethyl esters, alcohols, acids, aldehydes, sugar alcohols) showed an upward trend, fermentable sugars, amino acids, and C6-compounds were decreased. Terpenes presented stable behavior, with the exception of terpenols, which were increased at the beginning and were then decreased after the 5th day of fermentation.PMID:37375206 | DOI:10.3390/molecules28124653

The Transport and Uptake of Resveratrol Mediated via Glucose Transporter 1 and Its Antioxidant Effect in Caco-2 Cells

Wed, 28/06/2023 - 12:00
Molecules. 2023 Jun 6;28(12):4569. doi: 10.3390/molecules28124569.ABSTRACTResveratrol has anti-inflammatory, anti-cancer, and anti-aging pharmacological activities. There is currently a gap in academic research regarding the uptake, transport, and reduction of H2O2-induced oxidative damage of resveratrol in the Caco-2 cell model. This study investigated the role of resveratrol in the uptake, transport, and alleviation of H2O2-induced oxidative damage in Caco-2 cells. In the Caco-2 cell transport model, it was observed that the uptake and transport of resveratrol (10, 20, 40, and 80 μM) were time dependent and concentration dependent. Different temperatures (37 °C vs. 4 °C) could significantly affect the uptake and transportation of resveratrol. The apical to basolateral transport of resveratrol was markedly reduced by STF-31, a GLUT1 inhibitor, and siRNA intervention. Furthermore, resveratrol pretreatment (80 μM) improves the viability of Caco-2 cells induced by H2O2. In a cellular metabolite analysis combined with ultra-high performance liquid chromatography-tandem mass spectrometry, 21 metabolites were identified as differentials. These differential metabolites belong to the urea cycle, arginine and proline metabolism, glycine and serine metabolism, ammonia recycling, aspartate metabolism, glutathione metabolism, and other metabolic pathways. The transport, uptake, and metabolism of resveratrol suggest that oral resveratrol could prevent intestinal diseases caused by oxidative stress.PMID:37375124 | DOI:10.3390/molecules28124569

Detection of Antagonistic Compounds Synthesized by <em>Bacillus velezensis</em> against <em>Xanthomonas citri</em> subsp. <em>citri</em> by Metabolome and RNA Sequencing

Wed, 28/06/2023 - 12:00
Microorganisms. 2023 Jun 8;11(6):1523. doi: 10.3390/microorganisms11061523.ABSTRACTBiological control of plant diseases has gained attraction for controlling various bacterial diseases at a field trial stage. An isolated endophytic bacterium, Bacillus velezensis 25 (Bv-25), from Citrus species had strong antagonistic activity against Xanthomonas citri subsp. citri (Xcc), which causes citrus canker disease. When Bv-25 was incubated in Landy broth or yeast nutrient broth (YNB), the ethyl acetate extract of Landy broth exhibited higher levels of antagonistic activity against Xcc compared to that of YNB. Therefore, the antimicrobial compounds in the two ethyl acetate extracts were detected by high performance liquid chromatography-mass spectrometry. This comparison revealed an increase in production of several antimicrobial compounds, including difficidin, surfactin, fengycin, and Iturin-A or bacillomycin-D by incubation in Landy broth. RNA sequencing for the Bv-25 grown in Landy broth were performed, and the differential expressions were detected for the genes encoding the enzymes for the synthesis of antimicrobial compounds, such as bacilysin, plipastatin or fengycin, surfactin, and mycosubtilin. Combination of metabolomics analysis and RNA sequencing strongly suggests that several antagonistic compounds, especially bacilysin produced by B. velezensis, exhibit an antagonistic effect against Xcc.PMID:37375024 | DOI:10.3390/microorganisms11061523

Multi-Omics Analysis Demonstrates the Critical Role of Non-Ethanolic Components of Alcoholic Beverages in the Host Microbiome and Metabolome: A Human- and Animal-Based Study

Wed, 28/06/2023 - 12:00
Microorganisms. 2023 Jun 5;11(6):1501. doi: 10.3390/microorganisms11061501.ABSTRACTIt is known that alcoholic beverages alter the human gut microbiome. This study focused on the potential impact of non-ethanolic ingredients in whisky on the gut bacteriome. A pilot study was carried out on 15 whisky drinkers, 5 rice beer drinkers, and 9 non-drinkers to determine the effect of alcoholic beverages on the host microbiome and metabolome. Additionally, a mouse model was used to assess the differential impact of three whisky brands (each with an equal ethanol concentration). The results indicate that the non-ethanolic components have an impact on the gut microbiome, as well as on the metabolites in blood and feces. The amount of Prevotella copri, a typical core Indian gut bacterium, decreased in both the human and mouse groups of whisky type 1, but an increase in abundance of Helicobacteriaceae (p = 0.01) was noticed in both groups. Additionally, the alcohol-treated cohorts had lower levels of short-chain fatty acids (SCFAs), specifically butyric acid, and higher amounts of lipids and stress marker IL1-ß than the untreated groups (p = 0.04-0.01). Furthermore, two compounds, ethanal/acetaldehyde (found in all the whisky samples) and arabitol (unique to whisky type 1), were tested in the mice. Similar to the human subjects, the whisky type 1 treated mouse cohort and the arabitol-treated group showed decreased levels of Prevotella copri (p = 0.01) in their gut. The results showed that non-ethanolic compounds have a significant impact on host gut bacterial diversity and metabolite composition, which has a further vital impact on host health. Our work further emphasizes the need to study the impact of non-ethanolic ingredients of alcoholic beverages on host health.PMID:37375003 | DOI:10.3390/microorganisms11061501

Anti-Hypercholesterolemia Effects of Edible Seaweed Extracts and Metabolomic Changes in Hep-G2 and Caco-2 Cell Lines

Wed, 28/06/2023 - 12:00
Life (Basel). 2023 Jun 5;13(6):1325. doi: 10.3390/life13061325.ABSTRACTHypercholesterolemia is a major risk for the development of cardiovascular diseases (CVDs), the main cause of mortality worldwide, and it is characterized by high levels of circulating cholesterol. The drugs currently available for hypercholesterolemia control have several side effects, so it is necessary to develop new effective and safer therapies. Seaweeds serve as sources of several bioactive compounds with claimed beneficial effects. Eisenia bicyclis (Aramé) and Porphyra tenera (Nori) are edible seaweeds that were previously recognized as rich in bioactive compounds. In the present study, we aim to evaluate the anti-hypercholesterolemia effect of these two seaweed extracts and their health potential. Both extracts, but more efficiently Aramé extract, have liver 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) inhibitory activity as well as the capability to reduce approximately 30% of cholesterol permeation through human Caco-2 cells by simulating the intestinal lining, which is a target for hypercholesterolemia treatments. An untargeted metabolomic assay on human intestinal Caco-2 and liver Hep-G2 cell lines exposed to Aramé and Nori extracts revealed changes in the cells' metabolism, indicating the extracts' health beneficial effects. The metabolic pathways affected by exposure to both extracts were associated with lipid metabolism, such as phospholipids, and fatty acid metabolism, amino acid pathways, cofactors, vitamins, and cellular respiration metabolism. The effects were more profound in Aramé-treated cells, but they were also observed in Nori-exposed cells. The metabolite modifications were associated with the protection against CVDs and other diseases and to the improvement of the cells' oxidative stress tolerance. The results obtained for the anti-hypercholesterolemia properties, in addition to the revelation of the positive impact on cell metabolism, offer an important contribution for further evaluation of these seaweed extracts as functional foods or for CVD prevention.PMID:37374108 | DOI:10.3390/life13061325

Metabolomics in Acute Kidney Injury: The Clinical Perspective

Wed, 28/06/2023 - 12:00
J Clin Med. 2023 Jun 16;12(12):4083. doi: 10.3390/jcm12124083.ABSTRACTBACKGROUND: Acute kidney injury (AKI) affects increasing numbers of hospitalized patients worldwide. The diagnosis of AKI is made too late in most individuals since it is still based on dynamic changes in serum creatinine. In recent years, new AKI biomarkers have been identified; however, none of these can reliably replace serum creatinine yet. Metabolomic profiling (metabolomics) allows the concomitant detection and quantification of large numbers of metabolites from biological specimens. The current article aims to summarize clinical studies on metabolomics in AKI diagnosis and risk prediction.METHODS: The following databases were searched for references: PubMed, Web of Science, Cochrane Library, and Scopus, and the period lasted from 1940 until 2022. The following terms were utilized: 'AKI' OR 'Acute Kidney Injury' OR 'Acute Renal Failure' AND 'metabolomics' OR 'metabolic profiling' OR 'omics' AND 'risk' OR 'death' OR 'survival' OR 'dialysis' OR 'KRT' OR 'kidney replacement therapy' OR 'RRT' OR 'renal replacement therapy' OR 'recovery of kidney function' OR 'renal recovery' OR 'kidney recovery' OR 'outcome'. Studies on AKI risk prediction were only selected if metabolomic profiling allowed differentiation between subjects that fulfilled a risk category (death or KRT or recovery of kidney function) and those who did not. Experimental (animal-based) studies were not included.RESULTS: In total, eight studies were identified. Six studies were related to the diagnosis of AKI; two studies were performed on metabolic analysis in AKI risk (death) prediction. Metabolomics studies in AKI already helped to identify new biomarkers for AKI diagnosis. The data on metabolomics for AKI risk prediction (death, KRT, recovery of kidney function), however, are very limited.CONCLUSIONS: Both the heterogenous etiology and the high degree of pathogenetic complexity of AKI most likely require integrated approaches such as metabolomics and/or additional types of '-omics' studies to improve clinical outcomes in AKI.PMID:37373777 | DOI:10.3390/jcm12124083

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