Metabolomic Analysis

Heart failure (HF) with preserved ejection fraction (HFpEF) is increasingly recognized as an important clinical entity. Preclinical studies have shown differences in the pathophysiology between HFpEF and HF with reduced ejection fraction (HFrEF). Therefore, we hypothesized that a systematic metabolomic analysis would reveal a novel metabolomic fingerprint of HFpEF that will help understand its pathophysiology and assist in establishing new biomarkers for its diagnosis.

This graph represents the differential expression of the 252 bacteriocyte enriched genes (relative to the midgut) in the whole gut tissue of aposymbiotic flies (x-axis) and trypanosome infected flies (y-axis). Data point sizes represent the expression level (Log2 of counts per million (CPM) values) in the bacteriome-specific transcriptome. Transcripts were categorized as significantly up- or down-regulated if they were scored as differentially expressed by edgeR analysis with a P-value of less than 0.05 and a false discovery rate (FDR) score of less than 0.05 (see also Table S6 for detailed annotations).

Insects with restricted diets rely on obligate microbes to fulfil nutritional requirements essential for biological function. Tsetse flies, vectors of African trypanosome parasites, feed exclusively on vertebrate blood and harbour the obligate endosymbiont Wigglesworthia glossinidia. Without Wigglesworthia, tsetse are unable to reproduce. These symbionts are sheltered within specialized cells (bacteriocytes) that form the midgut-associated bacteriome organ. To decipher the core functions of this symbiosis essential for tsetse's survival, we performed dual-RNA-seq analysis of the bacteriome, coupled with metabolomic analysis of bacteriome and haemolymph collected from normal and symbiont-cured (sterile) females. Bacteriocytes produce immune regulatory peptidoglycan recognition protein (pgrp-lb) that protects Wigglesworthia, and a multivitamin transporter (smvt) that can aid in nutrient dissemination. Wigglesworthia overexpress a molecular chaperone (GroEL) to augment their transl...

Background Heart failure (HF) with preserved ejection fraction (HFpEF) is increasingly recognized as an important clinical entity. Preclinical studies have shown differences in the pathophysiology between HFpEF and HF with reduced ejection fraction (HFrEF). Therefore, we hypothesized that a systematic metabolomic analysis would reveal a novel metabolomic fingerprint of HFpEF that will help understand its pathophysiology and assist in establishing new biomarkers for its diagnosis.

Objective To interrogate the pathogenesis of intrauterine growth restriction (IUGR) and apply Artificial Intelligence (AI) techniques to multi-platform i.e. nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry (MS) based metabolomic analysis for the prediction of IUGR.

Ischemic stroke is one of the most common causes of death worldwide and is a major cause of acquired disability in adults. However, there is still a need for an effective drug for its treatment. Buyang Huanwu decoction (BHD), a traditional Chinese medicine (TCM) prescription, has long been used clinically to aid neurological recovery after stroke. To establish potential clinical indicators of BHD efficacy in stroke treatment and prognosis, we conducted a combined proteomic and metabolomic analysis of cerebrospinal fluid (CSF) samples in a mouse stroke model. CSF samples were obtained from male mice with acute ischemic stroke induced by middle cerebral ischemic/reperfusion (CI/R) injury, some of which were then treated with BHD. Label-free quantitative proteomics was conducted using nano-LC-MS/ MS on an LTQ Orbitrap mass and metabolomic analysis was performed using nanoprobe NMR and UHPLC-QTOF-MS. The results showed that several proteins and metabolites were present at significantly different concentrations in the CSF samples from mice with CI/R alone and those treated with BHD. These belonged to pathways related to energy demand, inflammatory signaling, cytoskeletal regulation, Wnt signaling, and neuroprotection against neurodegenerative diseases. In conclusion, our in silico data suggest that BHD treatment is not only protective but can also ameliorate defects in pathways affected by neurological disorders. These data shed light on the mechanism whereby BHD may be effective in the treatment and prevention of stroke-related neurodegenerative disease.

Ischemic stroke is one of the most common causes of death worldwide and is a major cause of acquired disability in adults. However, there is still a need for an effective drug for its treatment. Buyang Huanwu decoction (BHD), a traditional Chinese medicine (TCM) prescription, has long been used clinically to aid neurological recovery after stroke. To establish potential clinical indicators of BHD efficacy in stroke treatment and prognosis, we conducted a combined proteomic and metabolomic analysis of cerebrospinal fluid (CSF) samples in a mouse stroke model. CSF samples were obtained from male mice with acute ischemic stroke induced by middle cerebral ischemic/reperfusion (CI/R) injury, some of which were then treated with BHD. Label-free quantitative proteomics was conducted using nano-LC-MS/ MS on an LTQ Orbitrap mass and metabolomic analysis was performed using nanoprobe NMR and UHPLC-QTOF-MS. The results showed that several proteins and metabolites were present at significantly different concentrations in the CSF samples from mice with CI/R alone and those treated with BHD. These belonged to pathways related to energy demand, inflammatory signaling, cytoskeletal regulation, Wnt signaling, and neuroprotection against neurodegenerative diseases. In conclusion, our in silico data suggest that BHD treatment is not only protective but can also ameliorate defects in pathways affected by neurological disorders. These data shed light on the mechanism whereby BHD may be effective in the treatment and prevention of stroke-related neurodegenerative disease.

Spiroimines are a class of compounds produced by marine dinoflagellates with a wide range of toxicity and therapeutic potential. The smallest of the cyclic imines, portimine, is far less toxic than other known members in several animal models. Portimine has also been shown to induce apoptosis and reduce the growth of a variety of cancer cell lines at low nanomolar concentrations. In an effort to discover new spiroimines, the current study undertook a metabolomic analysis of cultures of cyclic imine-producing dinoflagellates, and a new analog of portimine was discovered in which the five-membered cyclic ether is open. Further scrutiny with human oral cavity squamous cell carcinoma (OCSCC) cell lines revealed that the open ring congener was less potent than portimine A but could still lead to the accumulation of apoptotic gene transcripts, fragment genomic DNA, and reduce cancer cell proliferation in the range of 100−200 nM.

Understanding cellular metabolism is a major challenge in current systems biology and has triggered extensive metabolomics research, which in most cases involves destructive analysis. However, the information obtainable only in a nondestructive manner will be required for accurately mapping the global structure of the organism's metabolic network at a given instant. Here we report that metabolic pathways can be explored in vivo by mixed stable isotopelabeled Raman microspectroscopy in conjunction with multivariate curve resolution analysis. As a model system, we studied ergosterol biosynthesis in single living fission yeast cells grown in mixtures of normal and 13 C-labeled glucose as the sole carbon source. The multivariate spectral data analysis of space-resolved Raman spectra revealed the intrinsic spectra and relative abundances of all isotopomers of ergosterol whose carbon atoms in the 5,7-diene moiety of the sterol skeleton are either partly or fully substituted with 13 C. Our approach is applicable to other metabolites and will earn a place in the toolbox of metabolomic analysis.

Introduction Grapevine protection is an important issue in viticulture. To reduce pesticide use, sustainable disease control strategies are proposed, including a promising alternative method based on the elicitor-triggered stimulation of the grapevine natural defense responses. However, detailed investigations are necessary to characterize the impact of such defense induction on the primary metabolism. Objectives Our aim was to use a metabolomics approach to assess the impact on grapevine of different elicitors dependent on the salicylic acid (SA) and/or jasmonic acid (JA) pathway. For this purpose, leaves of grapevine foliar cuttings were treated with methyl jasmonate, acibenzolar-S-methyl or phosphonates. Methods According to the elicitor, common and discriminating metabolites were elucidated using 1 H NMR measurements and principal component analysis. Results A wide range of compounds including carbohydrates, amino acids, organic acids, phenolics and amines were identified. The score plots obtained by combining PC1 versus PC2 and PC1 versus PC3 allowed a clear separation of samples, so metabolite fingerprinting showed an extensive reprogramming of primary metabolic pathways after elicitation. Conclusion The methods applied were found to be accurate for the rapid determination and differential characterization of plant samples based on their metabolic composition. These investigations can be very useful because the application of plant defense stimulators is gaining greater importance as an alternative strategy to pesticides in the vineyard.

Background:  A dietary supplement developed in Japan, OM-X®, is the result of extended fermentation of dozens of edible vegetables, fruits, seaweeds, mushrooms, and includes 12 strains of Lactic acid bacteria (LAB) and Bifidobacterium. The supplements powerful antioxidant, anti-inflammatory properties, and regulatory effect on metabolizing liver enzymes have been reported.
Senescence marker protein-30 (SMP30)/Gluconolactonase (GNL) Knockout (KO) mice, which lack the ability to biosynthesize Vitamin C (VC) in the body, were used in this study. The mice exhibited decreased antioxidant capacity in the blood, decreased liver function, and had poor hair growth due to the VC deficiency. The aim of the study was to examine the effects of oral administration of OM-X® on VC deficiency from the viewpoint of comprehensive analysis of liver metabolism.
Methods: SMP30/GNL KO and C57BL/6 (WT) mice were used in this study. The KO mice were divided into the following three groups: VC non-administration (VC minus), VC (0.15 W/V %) administration (VC plus), and no VC plus 0.6% OM-X® administration (OM-X®) groups (n = 6 per group). Mice were kept under experimental conditions until they were euthanized at week 8 (age: 16 weeks old). Body weight and plasma biochemistry levels were measured.. Metabolomic analyses of harvested livers were performed.
Results: In the VC minus group, a significant decrease in body weight and significant changes in plasma biochemistry parameters compared with the WT control or VC plus groups was observed. In the OM-X® group, significant suppression or a suppressive trend was observed compared with the VC minus group. Liver metabolomic analysis showed that total adenylate levels decreased in the VC minus group, whereas OM-X® administration significantly elevated total adenylate levels. In addition, the ratios of Glutathione (GSH)/ Glutathione disulfide (GSSG) and S-adenosylmethionine (SAM)/ S-adenosyl-L-homocysteine (SAH) and total glutathione levels were all significantly higher in the OM-X® group than in the VC minus group. The glycerol 3-phosphate/ Dihydroxyacetone phosphate (DHAP) and putrescine/spermidine ratios were elevated in the VC minus group, whereas it was significantly lower in the OM-X® group.
Conclusion: The results showed that in SMP30/GNL KO mice, some organ damage may have occurred during VC deficiency, as indicated by weight loss, hepatic injury, and changes in triglyceride-related markers. OM-X® had an effect on energy charge maintenance by elevating total adenylate levels, inducing antioxidant capacity via enhancing glutathione levels, and promoting protein synthesis, including polyamine synthesis. Altogether, the results revealed that OM-X® prevents the adverse biological changes caused by VC deficiency in SMP30/GNL KO mice.