Metabolic Profiling Analysis Research Articles

Untargeted and spatial-resolved metabolomics characterize serum and tissue-specific metabolic reprogramming in acute kidney injury

BackgroundAcute kidney injury (AKI) is one of the most common clinical emergencies characterized by rapid progression, difficulty in early diagnosis, and high mortality. Currently, there are no effective AKI early diagnostic methods and treatments. Therefore, identifying new mechanisms of AKI have become urgent for development new targets for early diagnosis and treatment of AKI in the current clinical setting. MethodsIn this study, systematic analysis and comparison of serum metabolic profiles of clinical AKI patients, chronic kidney disease (CKD) patients, and healthy subjects were performed using untargeted metabolomics. Moreover, the first spatial metabolomic analysis of kidney tissues in an AKI mouse model using MALDI-TOF MS technology was conducted. Differentially expressed metabolites were identified using a comprehensive, publicly available database. The metabolic data obtained were evaluated using principal component analysis, (orthogonal) partial least squares discriminant analysis, and metabolic pathway analysis to explore the unique serum metabolic profile of the patients, as well as to characterize the spatial distribution of differential metabolites in the kidneys of AKI mice. ResultsSignificant changes in the metabolite levels of amino acids, carnitine, and lipids were observed in the AKI and CKD groups versus the healthy population, suggesting that kidney injury may lead to abnormalities in various metabolic pathways, such as amino acids, fatty acids, and lipids. The significant difference between the AKI and CKD groups were found for the first time in these indexes including amino acid, carnitine, fatty acid, and lipid levels. Additionally, spatial metabolomics results revealed that amino acid, carnitine, organic acid, and fatty acid metabolites were more likely significantly altered in the renal cortex, while lipid metabolites were both differentially distributed in the cortex and medulla of the AKI group. ConclusionAbnormalities in the serum metabolism of amino acids, carnitine, and lipids in patients with kidney diseases, such as AKI and CKD, are closely associated with the physiological dysfunction of kidney injury. Metabolic differences between patients with AKI and CKD were compared for the first time, showing that fatty acid oxidative inhibition was more severe in patients with AKI. Furthermore, spatial metabolomics has revealed metabolic reprogramming with tissue heterogeneity in AKI mice model. Our study provides valuable information in the molecular pathological features of AKI in the kidney tissues.

Podophyllotoxin-mediated neurotoxicity via the microbiota-gut-brain axis in SD rats based on the toxicological evidence chain (TEC) concept

Podophyllotoxin (PPT) is a naturally occurring aryltetralin lignan. However, its clinical application has been limited due to its neurotoxicity, the mechanism of which remains unclear. This study aimed to investigate the potential involvement of the microbiota-gut-brain (MGB) axis in PPT-induced neurotoxicity using the toxicological evidence chain concept. Our approach included behavioral testing in rats, evaluation of colon and hippocampal pathological changes, examination of proinflammatory factors, brain-gut peptides, and an in-depth analysis of gut microbiome and metabolic profiles. Our results demonstrated that PPT exposure compromised cognitive functions, induced damage to the colon and hippocampus, and increased intestinal permeability in rats. Furthermore, it elevated proinflammatory factors, particularly TNF-α and IL-6, while causing disruptions in the gut microbiota, favoring Escherichia-Shigella over Lactobacillus. Significant alterations in metabolic profiles in feces, serum, and hippocampus, particularly in tryptophan metabolism with a correlation to inflammatory factors and Escherichia-Shigella, were also observed. Our findings suggest that PPT promotes the enrichment of Escherichia-Shigella leading to inflammatory factor production and alterations in kynurenine metabolism in the hippocampus, potentially contributing to neurotoxicity. The study provides novel insights into the mechanistic pathways of PPT-induced neurotoxicity, emphasizing the role of the MGB axis and offering avenues for therapeutic interventions.

Metabolic Profiling Analysis Uncovers the Role of Carbon Nanoparticles in Enhancing the Biological Activities of Amaranth in Optimal Salinity Conditions

Enhancing the productivity and bioactivity of high-functional foods holds great significance. Carbon nanoparticles (CNPs) have a recognized capacity for boosting both plant growth and the efficacy of primary and secondary metabolites. Furthermore, while salinity diminishes plant growth, it concurrently amplifies the production of phytomolecules. To ensure the robust and sustainable production of nutritious food, it becomes essential to elevate biomolecule yield without compromising plant growth. Here, we assessed the CNPs priming on plant performance and metabolites of the glycophyte amaranth (Amaranthus hypochondriacus) sprouts at the threshold salinity (25 mM NaCl; i.e., salinity that does not reduce growth but enhances the metabolites of that plant). We measured growth parameters, pigment levels, and primary (carbohydrates, amino acids, organic acids, fatty acids) and secondary metabolites (phenolics, flavonoids, tocopherols). CNP priming significantly improved biomass accumulation (fresh and dry weight) and primary and secondary metabolites of amaranth sprouts. Increased photosynthetic pigments can explain these increases in photosynthesis. Enhanced photosynthesis induced carbohydrate production, providing a C source for producing bioactive primary and secondary metabolites. The priming effect of CNPs further enhanced the accumulation of essential amino acids, organic acids, unsaturated fatty acids, tocopherols, and phenolics at threshold salinity. The increase in bioactive metabolites under threshold salinity can explain the CNP priming impact on boosting the antioxidant activities (FRAP, DPPH, anti-lipid peroxidation, superoxide-anion-scavenger, hydroxyl-radical-scavenger, Fe-chelating and chain-breaking activity in aqueous and lipid phases) and antimicrobial activities against Gram-positive and Gram-negative bacteria and fungi. Overall, this study suggested that threshold salinity and CNP priming could be useful for enhancing amaranth sprouts’ growth and nutritional quality.

Identification and biosynthesis of plant papanridins, a group of novel oligomeric flavonoids

The discovery of novel flavonoids and elucidation of their biosynthesis are fundamental to understanding their roles in plants and their benefits for human and animal health. Here, we report a new pathway for polymerization of a group of novel oligomeric flavonoids in plants. We engineered red cells for discovering genes of interest involved in the flavonoid pathway and identified a gene encoding a novel flavanol polymerase (FP) localized in the central vacuole. FP catalyzes the polymerization of flavanols, such as epicatechin and catechin, to produce yellowish dimers or oligomers. Structural elucidation shows that these compounds feature a novel oligomeric flaven–flavan (FF) skeleton linked by interflavan–flaven and interflaven bonds, distinguishing them from proanthocyanidins and dehydrodicatechins. Detailed chemical and physical characterizations further confirmed the novel FFs as flavonoids. Mechanistic investigations demonstrated that FP polymerizes flavan-3-ols and flav-2-en-3-ol carbocation, forming dimeric or oligomeric flaven-4→8-flavans, which we term “papanridins.” Data from transgenic experiments, mutant analysis, metabolic profiling, and phylogenetic analyses show that the biosynthesis of papanridins is prevalent in cacao, grape, blueberry, corn, rice, Arabidopsis, and other species in the plant kingdom. In summary, our study discoveries a group of novel oligomeric flavonoids, namely papanridins, and reveals that a novel FP-mediated polymerization mechanism for the biosynthesis of papanridins in plants.

Comparative analysis of bile metabolic profile in patients with biliary obstruction complicated by Clonorchis sinensis infection.

Clonorchiasis is an important foodborne parasitic disease. However, eggs of Clonorchis sinensis (C. sinensis) cannot be detected in feces during biliary obstruction. Moreover, many diseases can cause biliary obstruction, such as gallstones, adenocarcinoma, cholangiocarcinoma and Ascaris lumbricoides infection. Therefore, it is of great significance to distinguish between patients of biliary obstruction and biliary obstruction with C. sinensis infection. A total of 48 biliary obstruction patients were enrolled, including 23 infected with C. sinensis (C. sinensis) (OB+C.s) and 25 non-infected subjects (OB). The bile samples were collected by endoscopic retrograde cholangiopancreatography and analyzed using ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-QTOF MS). Additionally, multivariate statistical analysis methods were employed to identify differential metabolites. Next, bile amino acid levels were determined by targeted metabolomics analysis. A total of 146 and 132 significant metabolites were identified in electrospray ionization (ESI)+ and ESI- modes, respectively. The levels of amino acids (asparagine, glutamate, ornithine) and polyamines (spermidine and spermine) were significantly changed. Targeted analysis showed that the levels of amino acids (such as L-arginine, L-glutamine, L-lysine, L-propionic, and L-tyrosine) were lower in OB+C.s patients compared to those in OB patients. Marked metabolic pathways were involved in "Glutathione metabolism", "Caffeine metabolism", "Alanine, aspartate and glutamate metabolism", "Arginine and proline metabolism", "Purine metabolism", "Beta-Alanine metabolism", and "D-glutamine and D-glutamate metabolism". These results show that there were significant differences between OB+C.s and OB patients, especially in amino acids. The metabolic signature and perturbations in metabolic pathways may help to better distinguish OB+C.s and OB patients.

Subchronic toxicity evaluation of Huobahuagen extract and plasma metabolic profiling analysis combined with conventional pathology methods.

Huobahua, namely, Tripterygium hypoglaucum (Levl.) Hutch, known as a traditional Chinese herbal medicine, especially its underground parts, has been widely developed into several Tripterygium agents for the treatment of rheumatoid arthritis and other autoimmune diseases. It has sparked wide public concern about its safety, such as multi-organ toxicity. However, the toxic characteristics and damage mechanism of Huobahuagen extract (HBHGE) remain unclear. In the present study, subchronic oral toxicity study of HBHGE (10.0g crude drug/kg/day for 12 weeks) was performed in male rats. Hematological, serum biochemical, and histopathological parameters, urinalysis, and plasma metabolic profiling were assessed. The single-dose subchronic toxicity results related to HBHGE exhibited obvious toxicity to the testis and epididymis of male rats. Furthermore, plasma metabolomics analysis suggested that a series of metabolic disorders were induced by oral administration of HBHGE, mainly focusing on amino acid (glutamate, phenylalanine, and tryptophan) metabolisms, pyrimidine metabolism, glutathione metabolism, and steroid hormone biosynthesis. Moreover, it appeared that serum testosterone in male rats treated with HBHGE for 12 weeks, decreased significantly, and was susceptible to the toxic effects of HBHGE. Taken together, conventional pathology and plasma metabolomics for preliminarily exploring subchronic toxicity and underlying mechanism can provide useful information about the reduction of toxic risks from HBHGE and new insights into the development of detoxification preparations.

Metabolomic and transcriptomic analyses provide insights into the flavonoid biosynthesis in sea buckthorn (Hippophae rhamnoides L.)

Sea buckthorn is a plant with similarities to both food and medicine, possessing abundant flavonoids. However, the metabolic mechanism of flavonoid synthesis in sea buckthorn is unclear. Here, we performed transcriptomic and metabolomic analyses to explore the metabolic characteristics of flavonoids in different sea buckthorn tissues and their associated gene regulatory networks. A total of 441 metabolites were identified from sea buckthorn by metabolomics (UPLC-MS/MS), which includes 104 flavonoids with significant differences in the relative contents among tissues. Transcriptomic analysis identified 75 genes involved in the flavonoid biosynthesis pathway. The weighted gene correlation network analysis revealed that the MEdarkgreen module showed a significant correlation with flavonoid compounds and comprised ten genes related to flavonoid synthesis. Overexpression of HrMYB114 resulted in a significant increase in flavonoid content and expression of structural genes in the flavonoid synthesis pathway in transgenic hairy roots. This study provided a comprehensive analysis of metabolic and transcriptomic profiles, which helped to investigate the molecular mechanism of flavonoid biosynthesis.

Protective effect and mechanism of styrax on ischemic stroke rats: metabonomic insights by UPLC-Q/TOF-MS analysis

Context Styrax is used for prevention and treatment of cerebrovascular diseases. However, the underlying mechanism remains unclear. Objective To elucidate styrax’s anti-ischemic stroke protective effects and underlying mechanisms. Materials and methods An ischemic-stroke rat model was established based on middle cerebral artery occlusion (MCAO). Sprague-Dawley rats were randomly assigned to the following groups (n = 10) and administered intragastrically once a day for 7 consecutive days: sham, model, nimodipine (24 mg/kg), styrax-L (0.1 g/kg), styrax-M (0.2 g/kg) and styrax-H (0.4 g/kg). Neurological function, biochemical assessment, and ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry (UPLC-Q/TOF-MS)-based serum metabonomics were used to elucidate styrax’s cerebral protective effects and mechanisms. Pearson correlation and western blot analyses were performed to verify. Results The addition of 0.4 g/kg styrax significantly reduced cerebral infarct volume and neurobehavioral abnormality score. Different doses of styrax also decrease MDA, TNF-α, IL-6, and IL-1β, and increase SOD and GSH-Px in ischemic-stroke rats (p < 0.05; MDA, p < 0.05 only at 0.4 g/kg dose). Biochemical indicators and metabolic-profile analyses (PCA, PLS-DA, and OPLS-DA) also supported styrax’s protective effects. Endogenous metabolites (22) were identified in ischemic-stroke rats, and these perturbations were reversible via styrax intervention, which is predominantly involved in energy metabolism, glutathione and glutamine metabolism, and other metabolic processes. Additionally, styrax significantly upregulated phosphorylated AMP-activated protein kinase and glutaminase brain-tissue expression. Conclusion Styrax treatment could ameliorate ischemic-stroke rats by intervening with energy metabolism and glutamine metabolism. This can help us understand the mechanism of styrax, inspiring more clinical application and promotion.

Integrated metabolic profiling and transcriptome analysis of Lonicera japonica flowers for chlorogenic acid, luteolin and endogenous hormone syntheses

The active ingredients of many medicinal plants are the secondary metabolites associated with the growth period. Lonicera japonica Thunb. is an important traditional Chinese medicine, and the flower development stage is an important factor that influences the quality of medicinal ingredients. In this study, transcriptomics and metabolomics were performed to reveal the regulatory mechanism of secondary metabolites during flowering of L. japonica. The results showed that the content of chlorogenic acid (CGA) and luteolin gradually decreased from green bud stage (Sa) to white flower stage (Sc), especially from white flower bud stage (Sb) to Sc. Most of the genes encoding the crucial rate-limiting enzymes, including PAL, C4H, HCT, C3′H, F3′H and FNSII, were down-regulated in three comparisons. Correlation analysis identified some members of the MYB, AP2/ERF, bHLH and NAC transcription factor families that are closely related to CGA and luteolin biosynthesis. Furthermore, differentially expressed genes (DEGs) involved in hormone biosynthesis, signalling pathways and flowering process were analysed in three flower developmental stage.

Metabolic Profiling Early Post-Allogeneic Haematopoietic Cell Transplantation in the Context of CMV Infection.

Immune dysfunction resulting from allogeneic haematopoietic stem cell transplantation (aHSCT) predisposes one to an elevated risk of cytomegalovirus (CMV) infection. Changes in metabolism have been associated with adverse outcomes, and in this study, we explored the associations between metabolic profiles and post-transplantation CMV infection using plasma samples collected 7-33 days after aHSCT. We included 68 aHSCT recipients from Rigshospitalet, Denmark, 50% of whom experienced CMV infection between days 34-100 post-transplantation. First, we investigated whether 12 metabolites selected based on the literature were associated with an increased risk of post-transplantation CMV infection. Second, we conducted an exploratory network-based analysis of the complete metabolic and lipidomic profiles in relation to clinical phenotypes and biological pathways. Lower levels of trimethylamine N-oxide were associated with subsequent CMV infection (multivariable logistic regression: OR = 0.63; 95% CI = [0.41; 0.87]; p = 0.01). Explorative analysis revealed 12 clusters of metabolites or lipids, among which one was predictive of CMV infection, and the others were associated with conditioning regimens, age upon aHSCT, CMV serostatus, and/or sex. Our results provide evidence for an association between the metabolome and CMV infection post-aHSCT that is independent of known risk factors.

Transcription and Metabolic Profiling Analysis of Three Discolorations in a Day of Hibiscus mutabilis.

In this study, we used combined transcriptomics and metabolomics to analyze the H. mutabilis cultivar's genetic and physiological mechanisms during three flower color transition periods (from white to pink, then from pink to red) within the span of one day. As a result, 186 genes were found to be significantly increased with the deepening of the H. mutabilis flower color; these genes were mainly involved in the expression of peroxidase 30, zinc finger protein, phosphate transporter PHO1, etc. In contrast, 298 genes were significantly downregulated with the deepening of H. mutabilis flower color, including those involved in the expression of probable O-methyltransferase 3, copper binding protein 9, and heat stress transcription factor A-6b. Some genes showed differential expression strategies as the flower color gradually darkened. We further detected 19 metabolites that gradually increased with the deepening of the H. mutabilis flower color, including L-isoleucine, palmitic acid, L-methionine, and (+)-7-isonitrobenzene. The content of the metabolite hexadecanedioate decreased with the deepening of the H. mutabilis flower color. Combined transcriptomics and metabolomics revealed that the metabolic pathways, including those related to anthocyanin biosynthesis, cysteine and methionine metabolism, and sulfur metabolism, appear to be closely related to H. mutabilis flower color transition. This study served as the first report on the genetic and physiological mechanisms of short-term H. mutabilis flower color transition and will promote the molecular breeding of ornamental cultivars of H. mutabilis.

The Role of the Bile Microbiome in Common Bile Duct Stone Development.

Common bile duct (CBD) stones are a health concern for 10-20% of individuals with symptomatic gallstones, leading to health complications and placing a burden on healthcare systems. This study was initiated to investigate the changes in microbiome compositions and the metabolic signature associated with CBD stones. The research approach integrated taxonomic and functional data with metabolomics data, complemented by in vivo experiments. In a single tertiary healthcare institution, a total of 25 patients were enrolled who had undergone endoscopic retrograde cholangiopancreatography (ERCP) between February 2019 and January 2021. We harvested DNA from bile samples acquired from these individuals. The amplification of the bacterial 16S rRNA gene V3-V4 region was conducted through polymerase chain reaction (PCR), followed by sequencing. We utilized QIIME2 for a comprehensive data analysis. Furthermore, we performed a metabolomic analysis of the bile samples using nuclear magnetic resonance (NMR) spectroscopy. For the assessment of functional gene enrichment, we employed MetaboAnalyst 5.0. Lastly, we executed in vivo experiments on C57BL/6 mice and undertook histological examinations of tissue samples. Out of the 25 study subjects, 17 underwent ERCP due to CBD stones (the CBD stone group), while the remaining 8 had the procedure for different reasons (the non-CBD stone group). An alpha diversity analysis showed a significantly greater microbial diversity in the bile samples of the non-CBD stone group (p < 0.01), and a beta diversity analysis confirmed the greater microbial compositional abundance in the gut microbiomes in this group (p = 0.01). A taxonomic analysis revealed that the abundances of Enterococcaceae and Enterococcus were higher in the bile microbiomes of the CBD stone group. A metabolic profile analysis showed that the acetate, formate, and asparagine levels were higher in the CBD stone group. A pathway enrichment analysis showed the metabolic pathways (Arginine and Proline Metabolism, Aspartate Metabolism, Glycine, and Serine Metabolism, and Ammonia Recycling pathways) that were associated with these differences. Preclinical experiments demonstrated systemic inflammation and extracellular trap formation in the CBD stone group. Our study highlights the importance of biliary dysbiosis and bile metabolites, specifically acetate and formate, in CBD stone development and progression. These findings have implications for the development of diagnostic and therapeutic strategies using microbiomes for CBD stones.

Characteristics of steroid hormones in systemic lupus erythematosus revealed by GC/MS-based metabolic profiling.

Systemic lupus erythematosus (SLE) is a systemic autoimmune disease with a remarkable predominance in female, suggesting that steroid hormones may be involved in the pathogenesis. However, steroid signature of SLE patients has not been fully explored. A metabolic profiling analysis based on gas chromatography/mass spectrometry (GC/MS) with high sensitivity and reproducibility was employed to comprehensively reveal SLE-specific steroid alterations. More than 70 kinds of steroids in urine were detected by gas chromatography/mass spectrometry (GC/MS) to reveal SLE-specific steroid alterations. Principle component analysis demonstrated that the steroid profile was obviously distinguished between patients with SLE and controls. A lower level of total androgens was observed in patients, and nine androgens [dehydroepiandrosterone (DHEA), testosterone, Etio, androsterone, βαβ-Diol, Epi-An, Epi-DHT, 16α-OH-DHEA, and A-Diol] underwent significant decrease. Moreover, patients with SLE exhibited a slightly higher level of total estrogens than controls, and three estrogens (17-Epi-E3, 17α-E2, and E3) were remarkably increased. Furthermore, we identified the elevation of two sterols (Lan and Chol), and the reduction of one corticoid (11-DeoxyF) and two progestins (5α-DHP and 11β-OH-Prog) in patients. In this study, metabolic signature of urinary steroids associated with SLE was comprehensively defined by GC/MS for the first time, and steroid metabolism disorders were found in patients with SLE, especially the conversion of androgens to estrogens. Our findings will provide new insights for a deeper understanding of the mechanism of steroid hormones in the pathogenesis of SLE and will help to unravel the reason of sexual disparity in SLE.

Plasma metabolomics by nuclear magnetic resonance reveals biomarkers and metabolic pathways associated with the control of HIV-1 infection/progression.

How the human body reacts to the exposure of HIV-1 is an important research goal. Frequently, HIV exposure leads to infection, but some individuals show natural resistance to this infection; they are known as HIV-1-exposed but seronegative (HESN). Others, although infected but without antiretroviral therapy, control HIV-1 replication and progression to AIDS; they are named controllers, maintaining low viral levels and an adequate count of CD4+ T lymphocytes. Biological mechanisms explaining these phenomena are not precise. In this context, metabolomics emerges as a method to find metabolites in response to pathophysiological stimuli, which can help to establish mechanisms of natural resistance to HIV-1 infection and its progression. We conducted a cross-sectional study including 30 HESN, 14 HIV-1 progressors, 14 controllers and 30 healthy controls. Plasma samples (directly and deproteinized) were analyzed through Nuclear Magnetic Resonance (NMR) metabolomics to find biomarkers and altered metabolic pathways. The metabolic profile analysis of progressors, controllers and HESN demonstrated significant differences with healthy controls when a discriminant analysis (PLS-DA) was applied. In the discriminant models, 13 metabolites associated with HESN, 14 with progressors and 12 with controllers were identified, which presented statistically significant mean differences with healthy controls. In progressors, the metabolites were related to high energy expenditure (creatinine), mood disorders (tyrosine) and immune activation (lipoproteins), phenomena typical of the natural course of the infection. In controllers, they were related to an inflammation-modulating profile (glutamate and pyruvate) and a better adaptive immune system response (acetate) associated with resistance to progression. In the HESN group, with anti-inflammatory (lactate and phosphocholine) and virucidal (lactate) effects which constitute a protective profile in the sexual transmission of HIV. Concerning the significant metabolites of each group, we identified 24 genes involved in HIV-1 replication or virus proteins that were all altered in progressors but only partially in controllers and HESN. In summary, our results indicate that exposure to HIV-1 in HESN, as well as infection in progressors and controllers, affects the metabolism of individuals and that this affectation can be determined using NMR metabolomics.

Evaluation of multifactorial digestive disorders in a dairy herd at different stages of lactation.

The manager of a large dairy herd (total confinement, total mixed ration feeding, 10130 kg milk sold per cow and year) requested a workup of a digestive disorder problem that had been ongoing for several years. The cows were at all stages of lactation, and the incidence proportion (events/100 cows at risk) of digestive disorders was approximately 5 to 10%. The clinical picture included an abnormal demeanor of varying severity and signs that differed among the cows. The latter included decreased milk production and physical activity, low body condition score, abnormal rumen motility and stratification, small intestinal dilatation, diarrhea, undigested fiber particles in the feces, fever and abdominal pain.The following approach was used to investigate the digestive disorder problem:1. The herd was assessed for production levels, housing requirements, feeding protocols and animal health management. The latter comprised evaluation of different animal scores, metabolic profile analysis, diagnosis of disease, culling records and slaughter data. The results revealed risk factors concerning the feeding and animal health monitoring, (e. g. in dry matter intake and silage quality management, disease detection and diagnosis in fresh cows). The assessment also identified a high occurrence of digestive disorders of unknown origin.2. Fifteen cows that represented the ongoing digestive disorder problem were selected to undergo clinical examination, hematological analysis, urinalysis, and ultrasonography of the ventral abdomen. The clinical examinations revealed different digestive disorders, which were mainly inflammatory in nature, in all the cows. Eight cows had localized reticuloperitonitis and 13 had left displaced abomasum with different degrees of displacement and adhesions between the abomasum and reticulum attributable to reticuloperitonitis.Our results revealed a multifactorial problem caused by several risk factors relating to animal health and feeding management protocols that resulted in different types of digestive disorders. The wires from damaged tires used to hold the pit silo tarps in place were identified as a possible traumatic cause of the reticuloperitonitis. Treatment, prevention and follow-up of the different conditions were discussed.

Artificial intelligence to improve ischemia prediction in rubidium positron emission tomography

Abstract Funding Acknowledgements Type of funding sources: None. Background Patients are referred to coronary artery disease (CAD) testing based on their pre-test probability (PTP). The available, easy-to-use risk prediction tools use three variables only and their diagnostic accuracy is limited. Artificial intelligence-based tools incorporate multiple variables and could improve PTP assessment due to the resulting combinatorial power. The factors in the AI model are not seen as independent individual values, but are recognized as patterns derived by a combinatorial analysis of the individual metabolic profile of each patient. Purpose We aimed to compare the diagnostic accuracy of a novel, artificial intelligence-based tool with commonly used PTP tools to predict ischemia. Methods Consecutive patients (n = 2417) referred for Rubidium-Position Emission Tomography (PET) were evaluated. PTP was calculated using the ESC 2013/2019 and ACC 2012/2021 guidelines, and a memetic pattern-based algorithm (MPA) model incorporating symptoms, vitals, ECG and laboratory findings. Five risk categories (very low to very high risk) were defined (&amp;lt;5%, 5–15%, 15–50%, 50–85%, &amp;gt;85%). Ischemia was defined as summed difference score (SDS) ≥2 on PET. Receiver operator characteristics were calculated and compared using the DeLong method. Results Known CAD, ischemia and scar (detected on PET) were present in 46.3%, 37.1% and 24.6%, respectively. The MPA model was most accurate to assess PTP of ischemia (AUC: 0.758, ESC 2013: 0.661, ESC 2019: 0.673, ACC 2012: 0.585, ACC 2021: 0.667, p&amp;lt;0.0001 each) as depicted in figure 1. The MPA’s sensitivity and negative predictive value to rule-out ischemia were 99.1% and 96.4%, respectively. The model allocated patients more evenly across risk groups, reduced the proportion of patients in the 15–85% range by 29% (ACC 2012) to 51% (ESC 2013), and was the only risk tool to correctly estimate ischemia prevalence in the very low risk group (table 1). Conclusion The MPA model significantly improved PTP assessment of ischemia. The MPA model has originally been derived on the endpoint coronary stenosis &amp;gt;50% and validated in three clinical studies (1–3). This study now confirms that the MPA model applied to the endpoint ischemia outperforms the current risk assessment tools and is most accurate to assess PTP. It enables clinicians to safely exclude ischemia based on readily available variables without advanced testing. It could be used to improve risk stratification of patients and to significantly reduce unnecessary non-invasive functional tests.

Metabolic profiling and expression analysis of key genetic factors in the biosynthetic pathways of antioxidant metabolites in mungbean sprouts.

Mungbeans (Vigna radiata L.), a major legume crop in Asia, contain higher amounts of functional substances than other legumes, such as catechin, chlorogenic acid, and vitexin. Germination can improve the nutritional value of legume seeds. Here, 20 functional substances were profiled in germinated mungbeans and the expression levels of the transcripts of key enzymes in targeted secondary metabolite biosynthetic pathways were identified. VC1973A, a reference mungbean elite cultivar, had the highest amount of gallic acid (99.93 ± 0.13 mg/100 g DW) but showed lower contents of most metabolites than the other genotypes. Wild mungbeans contained a large amount of isoflavones compared with cultivated genotypes, especially for daidzin, genistin and glycitin. The expression of key genes involved in biosynthetic pathways had significant positive or negative correlations with the target secondary metabolite contents. The results indicate that functional substance contents are regulated at the transcriptional level, which can be applied to improve the nutritional value of mungbean sprouts in molecular breeding or genetic engineering, and wild mungbeans are a useful resource to improve the quality of mungbean sprouts.

Physiological traits and metabolic profiles of contrasting rice cultivars under mild salinity stress during the seedling stage

This study aimed to investigate the effects of mild salinity stress on some physiological traits and metabolic profiles of rice during the seedling stage. Three rice cultivars differing in salinity stress response, ‘KDML105’ (salt-sensitive, local Thai jasmine rice), ‘IR29’ (salt-sensitive rice), and ‘Pokkali’ (salt-tolerant rice), were subjected to mild salinity stress by flood-irrigated with 40 mM NaCl. Salinity stress significantly reduces the growth and biomass of the sensitive cultivars. Electrolyte leakage in ‘KDML105’ and ‘IR29’ also increased significantly while the relative water content (RWC) decreased. However, salinity stress did not affect growth, electrolyte leakage, and RWC in the tolerant cultivar. Salinity significantly increased Na+/K+ ratio in shoot and root of all cultivars. Nevertheless, the increase of Na+/K+ ratio in ‘KDML105’ and ‘IR29’ was higher than that of ‘Pokkali’. The metabolic profile analysis of ‘KDML105’ showed that uracil, L-proline, malonate, L-carnitine, L-lactate, and guanine were significantly increased, but the levels of dihydroxyacetone and glycine were significantly decreased under salinity conditions. In ‘IR29’, L-proline, melatonin, gamma-aminobutyrate, and acetate were significantly increased. In ‘Pokkali’, however, sugar alcohols and sugars were slightly increased when exposed to salinity conditions. The metabolic pathway analysis revealed one involved metabolic pathway of ‘KDML105’: aminoacyl-tRNA biosynthesis, and three involved those of ‘IR29’: arginine proline metabolism, sulfur metabolism, and butanoate metabolism. The difference in the metabolic profile of the salt-tolerance salt-sensitive cultivars provides new insights to identify additional novel biomarkers that will help better understand the mechanisms for salt tolerance in rice.

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

Mulberry 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.

A Deep Learning Approach to Analyze NMR Spectra of SH-SY5Y Cells for Alzheimer’s Disease Diagnosis

The SH-SY5Y neuroblastoma cell line is often used as an in vitro model of neuronal function and is widely applied to study the molecular events leading to Alzheimer’s disease (AD). Indeed, recently, basic research on SH-SY5Y cells has provided interesting insights for the discovery of new drugs and biomarkers for improved AD treatment and diagnosis. At the same time, untargeted NMR metabolomics is widely applied to metabolic profile analysis and screening for differential metabolites, to discover new biomarkers. In this paper, a compression technique based on convolutional autoencoders is proposed, which can perform a high dimensionality reduction in the spectral signal (up to more than 300 times), maintaining informative features (guaranteed by a reconstruction error always smaller than 5%). Moreover, before compression, an ad hoc preprocessing method was devised to remedy the scarcity of available data. The compressed spectral data were then used to train some SVM classifiers to distinguish diseased from healthy cells, achieving an accuracy close to 78%, a significantly better performance with respect to using standard PCA-compressed data.