Orthogonal Projections To Latent Structures Discriminant Analysis Research Articles

Imbalance of multiple neurotransmitter pathways leading to depression-like behavior and cognitive dysfunction in the triple transgenic mouse model of Alzheimer disease.

Depression is among the most frequent psychiatric comorbid conditions in Alzheimer disease (AD). However, pharmacotherapy for depressive disorders in AD is still a big challenge, and the data on the efffcacy of current antidepressants used clinically for depressive symptoms in patients with AD remain inconclusive. Here we investigated the mechanism of the interactions between depression and AD, which we believe would aid in the development of pharmacological therapeutics for the comorbidity of depression and AD. Female APP/PS1/Tau triple transgenic (3×Tg-AD) mice at 24 months of age and age- and sex-matched wild-type (WT) mice were used. The shuttle-box passive avoidance test (PAT) were implemented to assess the abilities of learning and memory, and the open field test (OFT) and the tail suspension test (TST) were used to assess depression-like behavior. High-performance liquid chromatography coupled to tandem mass spectrometry (HPLC-MS/MS) was used to detect the level of neurotransmitters related to depression in the hippocampus of mice. The data was identified by orthogonal projections to latent structures discriminant analysis (OPLS-DA). Most neurotransmitters exert their effects by binding to the corresponding receptor, so the expression of relative receptors in the hippocampus of mice was detected using Western blot. Compared to WT mice, 3×Tg-AD mice displayed significant cognitive impairment in the PAT and depression-like behavior in the OFT and TST. They also showed significant decreases in the levels of L-tyrosine, norepinephrine, vanillylmandelic acid, 5-hydroxytryptamine, and acetylcholine, in contrast to significant increases in 5-hydroxyindoleacetic acid, L-histidine, L-glutamine, and L-arginine in the hippocampus. Moreover, the expression of the alpha 1a adrenergic receptor (ADRA1A), serotonin 1A receptor (5HT1A), and γ-aminobutyric acid A receptor subunit alpha-2 (GABRA2) was significantly downregulated in the hippocampus of 3×Tg-AD mice, while histamine H3 receptor (H3R) expression was significantly upregulated. In addition, the ratio of phosphorylated cAMP-response element-binding protein (pCREB) and CREB was significantly decreased in the hippocampus of 3×Tg-AD mice than WT mice. We demonstrated in the present study that aged female 3×Tg-AD mice showed depression-like behavior accompanied with cognitive dysfunction. The complex and diverse mechanism appears not only relevant to the imbalance of multiple neurotransmitter pathways, including the transmitters and receptors of the monoaminergic, GABAergic, histaminergic, and cholinergic systems, but also related to the changes in L-arginine and CREB signaling molecules.

Silver Nanoparticle Targets Fabricated Using Chemical Vapor Deposition Method for Differentiation of Bacteria Based on Lipidomic Profiles in Laser Desorption/Ionization Mass Spectrometry.

The global threat of numerous infectious diseases creates a great need to develop new diagnostic methods to facilitate the appropriate prescription of antimicrobial therapy. More recently, the possibility of using bacterial lipidome analysis via laser desorption/ionization mass spectrometry (LDI-MS) as useful diagnostic tool for microbial identification and rapid drug susceptibility has received particular attention because lipids are present in large quantities and can be easily extracted similar to ribosomal proteins. Therefore, the main goal of the study was to evaluate the efficacy of two different LDI techniques-matrix-assisted (MALDI) and surface-assisted (SALDI) approaches-in the classification of the closely related Escherichia coli strains under cefotaxime addition. Bacterial lipids profiles obtained by using the MALDI technique with different matrices as well as silver nanoparticle (AgNP) targets fabricated using the chemical vapor deposition method (CVD) of different AgNP sizes were analyzed by the means of different multivariate statistical methods such as principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), sparse partial least squares discriminant analysis (sPLS-DA), and orthogonal projections to latent structures discriminant analysis (OPLS-DA). The analysis showed that the MALDI classification of strains was hampered by interference from matrix-derived ions. In contrast, the lipid profiles generated by the SALDI technique had lower background noise and more signals associated with the sample, allowing E. coli to be successfully classified into cefotaxime-resistant and cefotaxime-sensitive strains, regardless of the size of the AgNPs. AgNP substrates obtained using the CVD method were used for the first time for distinguishing closely related bacterial strains based on their lipidomic profiles and demonstrate high potential as a future diagnostic tool for the detection of antibiotic susceptibility.

Combination of peak-picking and binning for NMR-based untargeted metabonomics study

In NMR-based untargeted metabolomic studies, 1H NMR spectra are usually divided into equal bins/buckets to diminish the effects of peak shift caused by sample status or instrument instability, and to reduce the number of variables used as input for the multivariate statistical analysis. It was noticed that the peaks near bin boundaries may cause significant changes in integral values of adjacent bins, and the weaker peak may be obscured if it is allocated in the same bin with intense peaks. Several efforts have been taken to improve the performance of binning. Here we propose an alternative method, named P-Bin, based on the combination of the classic peak-picking and binning procedures. The location of each peak defined by peak-picking is used as the center of the individual bin. P-Bin is expected to keep all spectral information associated with the peaks and significantly reduce the data size as the spectral regions without peaks are not considered. In addition, both peak-picking and binning are routine procedures, making P-Bin easy to be implemented. To verify the performance, two sets of experimental data from human plasma and Ganoderma lucidum (G. lucidum) extracts were processed using the conventional binning method and the proposed method, before the principal component analysis (PCA) and the orthogonal projection to latent structures discriminant analysis (OPLS-DA). The results indicate that the proposed method has improved both the clustering performance of PCA score plots and the interpretability of OPLS-DA loading plots, and P-Bin could be an improved version of data preparation for metabonomic study.

Differentiating Huangjiu with Varying Sugar Contents from Different Regions Based on Targeted Metabolomics Analyses of Volatile Carbonyl Compounds.

Huangjiu is one of the oldest alcoholic beverages in the world. It is usually made by fermenting grains, and Qu is used as a saccharifying and fermenting agent. In this study, we identified differential carbonyl compounds in Huangjiu with varying sugar contents from different regions. First, we developed and validated a detection method for volatile carbonyl compounds in Huangjiu, and for optimal extraction, 5 mL of Huangjiu and 1.3 g/L of O-(2,3,4,5,6-pentafluorobenzyl)hydroxylamine hydrochloride (PFBHA) were incubated at 45 °C for 5 min before extracting the volatile carbonyl compounds at 45 °C for 35 min. Second, the targeted quantitative analysis of 50 carbonyl compounds in Huangjiu showed high levels of Strecker aldehydes and furans. Finally, orthogonal projections to latent structures discriminant analysis (OPLS-DA) was used to differentiate between Huangjiu with different sugar contents, raw materials, and region of origin. A total of 19 differential carbonyl compounds (VIP > 1, p < 0.05) were found in Huangjiu with different sugar contents (semidry and semisweet Huangjiu), and 20 differential carbonyl compounds (VIP > 1, p < 0.05) were found in different raw materials for Huangjiu production (rice and nonrice Huangjiu). A total of twenty-two and eight differential carbonyl compounds, with VIP > 1 and p < 0.05, were identified in semidry and semisweet Huangjiu from different regions (Zhejiang, Jiangsu, Shanghai, and Fujian), respectively.

Lipidome, central carbon metabolites, and sleep rhythm in coronary heart disease with nontraditional risks: An exploratory pilot study

AimsAltered lipid, energy metabolism and sleep disorders had been linked with coronary heart disease (CHD), however, the metabolic signatures and sleep rhythm in non-obstructive coronary atherosclerosis-CHD remain unclear. This pilot study aims to investigate the lipidome and central carbon metabolites profiles and associated sleep characteristics among CHD patients without traditional risk factors. MethodsFrom January to July 2021, 15 CHD patients and 15 healthy controls were randomly selected from the cardiology unit of Zhongshan Hospital, Shanghai. A total of 464 lipids and 45 central carbon metabolites (CCM) were quantified in blood plasma. Metabolic signatures were selected through orthogonal projections to latent structures discriminant analysis (OPLS-DA) and principal component analysis (PCA) was conducted to link the profiles of identified metabolites with CHD risk, sleep patterns, cardiometabolic traits and cardiac electrophysiologic parameters. ResultsUsing OPLS-DA, we identified 40 metabolites (variable influence on projection >1) that were altered in CHD patients, with 38 lipids, including 25 triacylglycerols (TAGs), 8 diacylglycerols (DAGs), being elevated and two CCM metabolites (i.e., succinic acid and glycolic acid) being reduced. Using PCA, four principal components (PCs) were identified and associated with increased risk of CHD. Specifically, one standard unit increasement in the PC that was characterized by high levels of DAG (18:1) and low succinic acid and the PC that was characterized by high levels of two sphingomyelins [SM (26:0) and SM (24:0)] was associated with 21% [odds ratio (OR) = 1.21, 95% CI: 1.02,1.43] and 14% (OR = 1.14,1.02,1.29) increased risk of CHD, respectively. Further regression analyses confirmed that the identified metabolites and the four PCs were positively associated with TG and ALT. Interestingly, glycolic acid was negatively associated with sleep quality and PSQI. Participants with night sleep mode tended to have a high level of the identified lipids, especially FFA (20:4). ConclusionIn the present pilot study, our findings provide clues on alterations of lipid and energy metabolism in CHD patients without traditional risk factors, with multiple triacylglycerols and diacylglycerols metabolites seemingly elevated and certain nonlipids metabolites (e.g., succinic acid and glycolic acid) decreased in cases. Considering the limit sample size, further studies are warranted to confirm our results.

Quantification and Diversity Analyses of Major Glucosinolates in Conserved Chinese Cabbage (Brassica rapa L. ssp. pekinensis) Germplasms

The genebank at the National Agrobiodiversity Center (RDA-Genebank, Jeonju, Republic of Korea), conserves approximately 8000 germplasms of Brassica spp., of which Chinese cabbage (Brassica rapa L. ssp. pekinensis) is one of the major crops actively used as food in Northeast Asia, including Korea, as the main ingredient for kimchi. Glucosinolates are a major class of compounds in Chinese cabbage that are responsible for their distinctive flavor, and RDA-Genebank is constantly building a related database (DB) to select suitable germplasms required by consumers and provide resources for breeding programs. In this study, ten glucosinolates were analyzed in sixty Chinese cabbage germplasms. Six aliphatic glucosinolates were the major components, accounting for 85.00% to 91.98% of total glucosinolates in each germplasm. Among them, gluconapin (333.26 to 23,501.58 μmol∙kg-1 DW) was highly represented, followed by glucobrassicanapin (545.60 to 10,344.70 μmol∙kg-1 DW) and progoitrin (155.28 to 8536.51 μmol∙kg-1 DW). In addition, we selected germplasms with a high content of each studied glucosinolate. To analyze the diversity and distribution of glucosinolates among the studied germplasms, Pearson's correlation was performed, and the related results were interpreted through their biosynthetic pathways. The k-means clustering indicated four optimal clusters, which were confirmed through principal component analysis. Orthogonal projection to latent structure discriminant analysis (OPLS-DA) was also performed on the status (landrace and cultivar) and origin (Korea, China, Taiwan, and Japan) passport data of the germplasms, followed by the calculation of variable importance in the projection (VIP) values. These results are part of a continuous series of studies to analyze the glucosinolates of Brassica germplasms that are being conserved at RDA-Genebank. We aim to provide related results through a public platform accessible to everyone and thereby improve the distribution of Brassica germplasms.

Metabolomic profiling of overnight peritoneal dialysis effluents predicts the peritoneal equilibration test type

This study primarily aimed to evaluate whether peritoneal equilibration test (PET) results can be predicted through the metabolomic analysis of overnight peritoneal dialysis (PD) effluents. From a total of 125 patients, overnight PD effluents on the day of the first PET after PD initiation were analyzed. A modified 4.25% dextrose PET was performed, and the PET type was categorized according to the dialysate-to-plasma creatinine ratio at the 4-h dwell time during the PET as follows: high, high average, low average, or low transporter. Nuclear magnetic resonance (NMR)-based metabolomics was used to analyze the effluents and identify the metabolites. The predictive performances derived from the orthogonal projection to latent structure discriminant analysis (OPLS-DA) modeling of the NMR spectrum were estimated by calculating the area under the curve (AUC) using receiver operating characteristic curve analysis. The OPLS-DA score plot indicated significant metabolite differences between high and low PET types. The relative concentrations of alanine and creatinine were greater in the high transporter type than in the low transporter type. The relative concentrations of glucose and lactate were greater in the low transporter type than in the high transporter type. The AUC of a composite of four metabolites was 0.975 in distinguish between high and low PET types. Measured PET results correlated well with the total NMR metabolic profile of overnight PD effluents.

Abstract P207: Heterogeneity in Obesity in Relation to Related to Hypertension: Investigating the Role of Metabolic Pathways

Background: While obesity is associated with hypertension, there is heterogeneity in risk by weight status and we know little about whether and which metabolic pathways play a role in exacerbating differential risk across weight status categories. Methods: Fasting blood was collected at the Yr. 20 follow-up exam in the Coronary Artery Risk Development in Young Adults (CARDIA) Study. We conducted Ultra-High-Performance Liquid Chromatography high resolution mass spectrometry untargeted metabolomics in 1,769 individuals (ages 37-54, 58% female, 66% Black) classified as normotensive (no hypertension diagnosis or medication and SBP/DBP ≤ 130/80). We identified 7,522 metabolomic peaks and 8 principal components, which we included in a multivariable-adjusted proportional hazards model with an interaction term to assess whether the association between these metabolite measures and incident hypertension (HTN) varied by BMI. We then employed multivariate analysis using Orthogonal Projections to Latent Structures Discriminant Analysis (OPLS-DA) models using 70% of the sample within each obesity category to predict incident HTN at ten years in normal weight, overweight and obese groups based on the high dimensional metabolomics data. The remaining 30% of individuals in each obesity category were then used to evaluate the predictive value of the OPLS-DA models. Finally, we used mummichog software and Fisher’s exact tests, to conduct pathway enrichment analysis using data from both the individual peak and the OPLS-DA models. Results: We identified 723 metabolites where the association with long term risk of HTN differed by BMI, although none were statistically significant after adjustment for multiple comparisons. However, we found strong evidence of effect modification for metabolites via principal components and OPLS-DA. For the 3 rd principal component, which explained 5% of the variation across all peaks, we found a significant interaction with BMI (p-value = 0.0043). In external test data sets, OPLS-DA models predicted incident HTN relatively well in normal weight individuals (AUC of 77%) but not well in overweight and obese individuals (AUC of 49% and 56% respectively). We identified several pathways with a differential HTN risk by BMI shown in the OPLS-DA and proportional hazards models. In the phenylamine pathway there was substantial differentiation between those with and without incident HTN risk for normal weight only and across the lysine degradation pathway only for individuals who were obese. Conclusions: Our findings identify variation in hypertension risk by BMI, with suggestive evidence for a role of metabolomic pathways and combinations of metabolites. Overall, metabolites predicted 10-year incident hypertension very well in individuals of normal weight (but not obese status), indicating heterogeneity in hypertension risk by weight status.

Abstract P414: Modification of Diet-Metabolite Associations by Race and Sex in the Coronary Artery Risk Development in Young Adults Study

Objective: There is known heterogeneity in the relationship between diet quality and cardiovascular disease by self-reported race and sex, which may underlie inequities in cardiovascular health. Methods: We used data from 2,832 participants in the Coronary Artery Risk Development in Young Adults (CARDIA) study (2005-06, aged 37-55, 57% women, 45% Black). Using fasted blood samples, we obtained ultra-high-performance liquid chromatography high resolution mass spectrometry untargeted metabolomics and derived an a priori diet quality score using a validated diet history questionnaire, based on classification of 46 food groups with potential beneficial (n=20), adverse (n=13), or neutral (n=13) implications for cardiovascular health. We tested effect modification of associations between metabolites and diet quality by race and sex, separately, using multivariable-adjusted linear regression for 7,522 metabolite peaks, accounting for multiple comparisons, adjusted for batch, field center, demographics, lifestyle behaviors, total energy intake, hypertension and diabetes status, medication use, and BMI. We also used race- and sex-stratified multivariate Orthogonal Projections to Latent Structures Discriminant Analysis (OPLS-DA) analyses to examine how metabolites distinguish by diet quality 1 st and 4 th quartiles. We identified differing metabolite pathways by race and sex through biochemical pathway analysis (Mummichog version 1.0.10) from regression and OPLS-DA models. Results: In linear regression, race significantly modified metabolite-diet quality associations for 231 metabolite peaks; sex modified associations for 1 peak (FDR&lt;0.1). OPLS-DA pathway analysis identified cysteine, methionine, histidine, and alpha-linoleic metabolism and terpenoid backbone biosynthesis as significantly associated with diet quality in Black, but not White, race. Linear regression and OPLS-DA pathway analysis found differences (p&lt;0.05) in arginine biosynthesis by sex. OPLS-DA also found aminoacyl-tRNA and arginine biosynthesis, nitrogen, porphyrin, and chlorophyll metabolism were significantly associated with diet quality in men only. Conclusions: Metabolite pathways were statistically different by self-reported race, and to a lesser extent, by sex. Differences in metabolite-diet quality associations may reflect differences in contextual or social variables, and potentially, heterogeneity in metabolism.

Unraveling putative antiulcer phytoconstituents against Helicobacter pylori urease and human H+/K+-ATPase from Jacaranda mimosifolia using UPLC-MS/MS coupled to chemometrics and molecular docking

Globally peptic ulcer disease is a severe public health issue caused by an imbalance between the defensive and aggressive mechanisms. In the current study, the phytochemical components of various organs (leaf, fruit, seed, and bark) of Jacaranda mimosifolia D. Don collected from Egypt at different seasons and geographical locations were investigated. UPLC-MS/MS allows a tentative identification of 53 compounds where flavonoids, quinoids, and triterpenoids represented the major identified classes. Multivariate statistical analysis (principle component analysis (PCA), hierarchical cluster analysis (HCA)-heat map and orthogonal projection to latent structures-discriminate analysis (OPLS-DA), and accompanied coefficient plots) was applied to investigate in-between and within-class discrimination of the different organs of Jacaranda mimosifolia D. Don. The obtained extracts were tested for their inhibitory activities against Helicobacter pylori Urease and human H+/K+-ATPase enzymes. Fruit extract showed the highest inhibitory activity against H. pylori urease. Meanwhile, leaf extract was found to be more potent, against human H+/K+-ATPase. The results of the biological activities were further modeled using Partial Least Squares Regression (PLSR) analysis together with the quantitative data. The optimal number of latent variables was determined with the aid of Variables important for the projection (VIP) plot to remove the uninformative variables. The developed model was validated using root mean square error of calibration (RMSEC), cross-validation (RMSECV) and prediction (RMSEP). PLS model coefficient plots showed that flavonoids as isoquercitrin and rutin were the most contributing metabolites/biomarkers to the detected anti-urease and H+/K+-ATPase activities, respectively. Molecular docking analysis was utilized to gain more insights into the interaction modalities for the top fifteen metabolites obtained from the PLS model with the active sites of urease and H+/K+-ATPase enzymes. In agreement with the model coefficients, isoquercitrin and rutin were ranked as the top hits for the studied enzymes. To validate the results, in vitro inhibitory assays were performed on isoquercitrin and rutin. Potent inhibitory activities were determined against urease and H+/K+-ATPase with IC50 ± SD of 9.10 ± 0.75 µM and 0.61 ± 0.06 µM for isoquercitrin and rutin, respectively.

Decoding the Effect of Age on the Taste Perception of Chicken Breast Soup Based on LC-QTOF-MS/MS Combined with a Chemometric Approach.

A nontargeted fingerprinting approach combined with the chemometrics method and sensory analysis was used to assess the differences in taste-chemical compositions of chicken breast soup with different ages and their sensory qualities. The sensory evaluation results showed that the overall taste as well as the sourness, saltiness, and umami scores of the soup were increased with the age of chicken. Fifty-nine compounds were identified from four soup samples by liquid chromatography-tandem quadrupole time-of-flight mass spectrometry (LC-QTOF-MS/MS), and their total content was the highest in the 90 wk soup samples. Six upregulated compounds (carnosine, hypoxanthine, inosine, inosine 5'-monophosphate (5'-IMP), adenosine 5'-monophosphate (5'-AMP), and lactic acid) were identified as potential contributors to the taste characteristics of the 90 wk soup samples by orthogonal projections to latent structures-discriminant analysis (OPLS-DA). Additional experiments showed that 5'-AMP particularly contributed to the sourness of the soup, while carnosine contributed to the saltiness and umami of the soup.

Antimicrobial and antibiofilm activities of pomegranate peel phenolic compounds: Varietal screening through a multivariate approach

Pomegranates are rich in phenolic compounds and known for their antioxidant, anti-inflammatory, and anticancer properties. The highest concentration of these compounds is found in the peel (exocarp and mesocarp), which constitutes about 50% of the whole fresh fruit. These bioactive phytochemicals exhibit a broad spectrum of antimicrobial effects against both Gram-negative and Gram-positive bacteria, as well as fungi. In the present paper, the chemical composition and antimicrobial activity of the peel (exocarp and mesocarp) from seven Punica granatum varieties (Wonderful, Mollar de Elche, Primosole, Sassari 1, Sassari 2, Sassari 3, and Arbara Druci) grown in Sardinia (Italy) were evaluated. Polar phenols, flavonoids, condensed tannins, and anthocyanin contents were evaluated by extraction with water at 20 and 40 °C. Orthogonal projections to latent structures discriminant analysis (OPLS-DA) was used to characterize each variety according to the chemical composition of the pomegranate peel extracts (PPEs). The antimicrobial and antibiofilm activities of each PPE were further tested in vitro against Staphyloccocus aureus, Listeria monocytogenes, Salmonella bongori, Escherichia coli, Lacticaseibacillus casei and Limosilactobacillus reuteri. Gram-positive species were more sensitive than Gram-negative to the extracts tested. Antimicrobial activity was shown against S. aureus and L. monocytogenes strains, whereas less, even no activity was found against S. bongori and E. coli strains. The PPEs from Mollar de Elche, Primosole, and Sassari 3 showed the highest antimicrobial activities at concentrations that varied from 0.19 to 1.50 mg/mL, with biofilm activity being reduced by more than 70%. These activities were positively related to the punicalagin, flavonoid, and chlorogenic acid content of the extracts. Finally, regarding the pro-technological bacterial strains, La. casei and Li. reuteri showed very low, even no sensitivity to the used of the specific PPEs with high concentrations. This study proposes a formulation of pomegranate peel extract that valorizes agro-industrial waste in the context of sustainability and circular economy. Pomegranate extracts should be considered potential sources of natural, plant-derived antimicrobials, providing an alternative to artificial antimicrobial products.

Metabolomics as a Potential Chemotaxonomical Tool: Application on the Selected Euphorbia Species Growing Wild in Serbia.

Chemotaxonomy presents various challenges that need to be overcome in order to obtain valid and reliable results. Individual genetic and environmental variations can give a false picture and lead to wrong conclusions. Applying a holistic approach, based on multivariate data analysis, these challenges can be overcome. Thus, a metabolomics approach has to be optimized depending on the subject of research. We used 1H NMR-based metabolomics as a potential chemotaxonomic tool on the selected Euphorbia species growing wild in Serbia. Principal components analysis (PCA), soft independent modeling by class analogy (SIMCA) and Orthogonal Projections to Latent Structures Discriminant Analysis (OPLS-DA) were used to analyze obtained NMR data in order to reveal chemotaxonomic biomarkers. The standard protocol for plant metabolomics was optimized aiming to extract more specific metabolites, which are characteristic for the Euphorbia genus. The obtained models were validated, which revealed that variables unique for each species were associated with certain classes of molecules according to literature data. In E. salicifolia, acacetin-7-O-glycoside (not found before in the species) was detected, and the structure of the aglycone part was solved based on 2D NMR data. In the presented paper, we have shown that metabolomics can be successfully used in Euphorbia chemotaxonomy.

Current trends in ŒNO-NMR based metabolomics

Present work discusses strengths and limitations of two Nuclear Magnetic Resonance outliers obtained with a water-to-ethanol solvent multi pre saturation acquisition method, recently included in the Compendium of International Methods of Analysis of Wines and Musts, published as OIV-MA-AS316-01, and their accuracy for metabolomics analysis. Furthermore, it is also presented an alternative to produce more discriminant and sensitive NMR data matrices for metabolomics studies, comprising the use of a novel NMR acquisition strategy in wines, the double pulsed-field gradient echo (DPFGE) NMR scheme, with a refocusing band-selective uniform-response pure-phase selective pulse, for a selective excitation of the 5-10 ppm chemical shift range of wine samples, that reveals novel broad aromatic 1H resonances, directly associated to complex polyphenols. Both aromatics and full binned OIV-MA-AS316-01,as well as the selective 5-10 ppm DPFGE NMR outliers were statistically analyzed with diverse non-supervised Principal Component Analysis (PCA) and supervised Partial Least Squares -Discriminant Analysis (PLS-DA), sparse (sPLS-DA) least squares- discriminant analysis, and orthogonal projections to latent structures discriminant analysis (OPLS-DA). Supervised multivariate statistical analysis of DPFGE and aromatics’ binned OIV-MA-AS316-01NMR data have shown their robustness to broadly discriminate geographical origins and narrowly differentiate between different fermentation schemes of wines from identical variety and region.

Comparison of Balanites aegyptiaca parts: metabolome providing insights into plant health benefits and valorization purposes as analyzed using multiplex GC-MS, LC-MS, NMR-based metabolomics, and molecular networking.

Balanites aegyptiaca (L.) Delile (Zygophyllaceae), also known as the desert date, is an edible fruit-producing tree popular for its nutritional and several health benefits. In this study, multi-targeted comparative metabolic profiling and fingerprinting approaches were conducted for the assessment of the nutrient primary and secondary metabolite heterogeneity in different parts, such as leaves, stems, seeds, unripe, and ripe fruits of B. aegyptiaca using nuclear magnetic resonance (NMR), ultra-performance liquid chromatography (UPLC-MS), and gas chromatography mass-spectrometry (GC-MS) based metabolomics coupled to multivariate analyses and in relation to its cytotoxic activities. NMR-based metabolomic study identified and quantified 15 major primary and secondary metabolites belonging to alkaloids, saponins, flavonoids, sugars, and amino and fatty acids. Principal component analysis (PCA) of the NMR dataset revealed α-glucose, sucrose, and isorhamnetin as markers for fruit and stem and unsaturated fatty acids for predominated seeds. Orthogonal projections to latent structure discriminant analysis (OPLS-DA) revealed trigonelline as a major distinctive metabolite in the immature fruit and isorhamnetin as a major distinct marker in the mature fruit. UPLC-MS/MS analysis using feature-based molecular networks revealed diverse chemical classes viz. steroidal saponins, N-containing metabolites, phenolics, fatty acids, and lipids as the constitutive metabolome in Balanites. Gas chromatography-mass spectroscopy (GC-MS) profiling of primary metabolites led to the detection of 135 peaks belonging to sugars, fatty acids/esters, amino acids, nitrogenous, and organic acids. Monosaccharides were detected at much higher levels in ripe fruit and disaccharides in predominate unripe fruits, whereas B. aegyptiaca vegetative parts (leaves and stem) were rich in amino acids and fatty acids. The antidiabetic compounds, viz, nicotinic acid, and trigonelline, were detected in all parts especially unripe fruit in addition to the sugar alcohol d-pinitol for the first time providing novel evidence for B. aegyptiaca use in diabetes. In vitro cytotoxic activity revealed the potential efficacy of immature fruit and seeds as cytotoxic agents against human prostate cancer (PC3) and human colorectal cancer (HCT-116) cell lines. Collectively, such detailed profiling of parts provides novel evidence for B. aegyptiaca medicinal uses.

Differences in Metabolomic Profiles by Birthplace in Mexican-Origin Hispanic Men Who Participated in a Weight Loss Lifestyle Intervention.

Birthplace, as a proxy for environmental exposures (e.g., diet), may influence metabolomic profiles and influence risk of cancer. This secondary analysis investigated metabolomic profile differences between foreign and U.S.-born Mexican-origin (MO) Hispanic men to shed light on potential mechanisms through which foreign- and U.S.-born individuals experience differences in cancer risk and risk factors. Plasma samples from MO Hispanic men (N = 42) who participated in a previous lifestyle intervention were collected pre-and post-intervention. Metabolomic profiles were characterized from samples using ultra performance liquid chromatography-quadrupole time of flight mass spectrometry (UPLC-QTOF). Models were visualized using supervised orthogonal projections to latent structures-discriminant analysis (OPLS-DA). Progenesis QI was used for peak integration and metabolite identification. Plasma metabolomic profiles differed between foreign- and U.S.-born pre-intervention (R2 = .65) and post-intervention (R2 = .62). Metabolomic profiles differed pre- versus post-intervention (R2 = .35 and R2 = .65) for the foreign- and U.S.-born group, respectively. Both endogenous metabolites and dietary components characterized differences between foreign- and U.S.-born participants pre- and post-intervention. Plasma metabolomic profiles from MO Hispanic men differed by birthplace. These results advance our understanding of relevant exposures that may affect cancer risk among MO Hispanic men born abroad or in the United States.

Metabolic analysis of the regulatory mechanism of sugars on secondary flowering in Magnolia

BackgroundMagnolia, a traditional and important ornamental plant in urban greening, has been cultivated for about 2000 years in China for its elegant flower shape and gorgeous flower color. Most varieties of Magnolia bloom once a year in spring, whereas a few others, such as Magnolia liliiflora Desr. ‘Hongyuanbao’, also bloom for the second time in summer or early autumn. Such a twice flowering trait is desirable for its high ornamental value, while its underlying mechanism remains unclear.MethodsParaffin section was used to show the flowering time and phenotypic changes of M. liliiflora ‘Hongyuanbao’ during the twice flowering periods from March 28 to August 25, 2018. Gas chromatography-mass spectrometry (GC-MS) was then performed to explore the chemical metabolites through the twice flower bud differentiation process in ‘Hongyuanbao’, and the metabolites were screened and identified by orthogonal projection to latent structures discriminant analysis (OPLS-DA). Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis (KEGG) was used to reveal the relationship between the sugar metabolites and twice-flowering characteristic. To further investigate the potential role of sucrose and trehalose on flowering regulation of ‘Hongyuanbao’, the plants once finished the spring flowering were regularly sprayed with sucrose and trehalose solutions at 30 mM, 60 mM, and 90 mM concentrations from April 22, 2019. The flower bud differentiation processes of sprayed plants were observed and the expression patterns of the genes involved in sucrose and trehalose metabolic pathways were studied by quantitative reverse transcription PCR (qRT-PCR).ResultsIt showed that ‘Hongyuanbao’ could complete flower bud differentiation twice in a year and flowered in both spring and summer. The metabolites of flower bud differentiation had a significant variation between the first and second flower buds. Compared to the first flower bud differentiation process, the metabolites in the sucrose and trehalose metabolic pathways were significantly up-regulated during the second flower bud differentiation process. Besides that, the expression levels of a number of trehalose-6-phosphate synthase (TPS) genes including MlTPS1, MlTPS5, MlTPS6, MlTPS7 and MlTPS9 were substantially increased in the second flower differentiation process compared with the first process. Exogenous treatments indicated that compared to the control plants (sprayed with water, CK), all three concentrations of trehalose could accelerate flowering and the effect of 60 mM concentration was the most significant. For the sucrose foliar spray, only the 60 mM concentration accelerated flowering compared with CK. It suggested that different concentration of trehalose and sucrose might have different effects. Expression analysis showed that sucrose treatment increased the transcription levels of MlTPS5 and MlTPS6, whereas trehalose treatment increased MlTPS1, showing that different MlTPS genes took part in sucrose and trehalose metabolic pathways respectively. The expression levels of a number of flowering-related genes, such as MlFT, MlLFY, and MlSPL were also increased in response to the sprays of sucrose and trehalose.ConclusionsWe provide a novel insight into the effect of sucrose and trehalose on the flowering process in Magnolia. Under the different sugar contents treatments, the time of flower bud differentiation of Magnolia was advanced. Induced and accelerated flowering in response to sucrose and trehalose foliar spray, coupled with elevated expression of trehalose regulatory and response genes, suggests that secondary flower bud formation is a promoted by altered endogenous sucrose and trehalose levels. Those results give a new understanding of sucrose and trehalose on twice-flowering in Magnolia and provide a preliminary speculation for inducing and accelerating the flowering process in Magnolia.

Untargeted 1 H-NMR Metabolome of Celery During Fusarium Wilt: Implications for Vegetable Quality.

Celery is a vegetable widely consumed as a condiment to prepare diverse dishes around the world. Nevertheless, this plant is susceptible to the attack of several phytopathogens including those of the Fusarium genus which is translated into devastating losses for the production chain. Herein we report on the metabolic changes produced during the celery wilt caused by Fusarium oxysporum which was determined through untargeted 1 H-NMR metabolomics. The changes in the metabolite content of celery were measured at 16, 24, and 32 days post-inoculation using viable conidia obtained from the native F. oxysporum strain FO3. Our results demonstrated that the parasitic activity of the fungus reduced the endogenous levels of free sugars (fructose, galactose, glucose isomers, mannose, Myo-inositol, mannitol, and sucrose) amino acids (alanine, aspartate GABA, glutamate, glutamine, histidine, isoleucine, leucine, methionine, proline, threonine, tyrosine, and valine), nucleosides (adenosine, cytidine, guanosine, and uridine) and organic acids (citric acid, fumaric acid, malic acid, and succinic acid). Interestingly, the levels of tyrosine and tryptophan were triggered as a consequence of F. oxysporum infection. This tendency was correlated with an increase in the levels of chlorogenic acid, apiin, and apigenin derivatives, suggesting their involvement in the chemical defense of celery against fungal colonization. According to principal component analysis (PCA) and Orthogonal Projections to Latent Structures Discriminant Analysis (OPLS-DA) methanol was the main differential metabolite and it was considered as a new chemical marker associated with F. oxysporum infection. Our results demonstrate that infected celery plants dramatically reduced their nutritional and nutraceutical contents during Fusarium wilt after 32 days post-inoculation. However, these findings also suggest that the phenylpropanoid pathway is strongly related with the chemical defense of celery against F. oxysporum.

Comparison of Lycium barbarum fruits polysaccharide from different regions of China by acidic hydrolysate fingerprinting-based HILIC-ELSD-ESI-TOF-MS combined with chemometrics analysis.

Lycium barbarum is an edible fruit widely used in herbal medicines and as a functional food. Polysaccharide is one of the most important active ingredients. Only L. barbarum grown in the Ningxia region of China are officially recognised as suitable for use in traditional Chinese medicine, but the systematic comparison of L. barbarum polysaccharide between Ningxia and the other growing regions of China has been rarely reported. To compare the difference of L. barbarum polysaccharide from different grown regions of China. A chemical fingerprint of L. barbarum polysaccharide hydrolysates was established based on controlled acidolysis combined with hydrophilic interaction liquid chromatography-evaporative light scattering detection-electrospray ionisation-time-of-flight-mass spectrometry (HILIC-ELSD-ESI-TOF-MS). Then, it was employed for the comparison of L. barbarum samples from different geographical origins of China combined with chemometrics analysis. Six monosaccharides [rhamnose (Rha), xylose (Xyl), arabinose (Ara), mannose (Man), glucose (Glu), galactose (Gal)] were qualitatively and quantitatively determined and four glycoconjugates were preliminarily identified from the hydrolysates. Content determination for the polysaccharide and monosaccharide indicated obvious geographical features. The HILIC-ELSD fingerprint combined with partial least squares-discriminant analysis (PLS-DA) was able to differentiate L. barbarum samples from Ningxia, Xinjiang, Gansu and Qinghai regions with 89.19% classification accuracy. Orthogonal projection to latent structure discriminant analysis (OPLS-DA) was able to differentiate between samples from Ningxia and those from the other three growing regions, polysaccharide and Ara were the potential chemical markers. These findings form the basis of a reliable method to trace the region of origin of L. barbarum sample and thereby, improve the quality control of L. barbarum therapeutic polysaccharides.

Detection of Pork in Beef Meatballs Using LC-HRMS Based Untargeted Metabolomics and Chemometrics for Halal Authentication

Adulteration of high-quality meat products using lower-priced meats, such as pork, is a crucial issue that could harm consumers. The consumption of pork is strictly forbidden in certain religions, such as Islam and Judaism. Therefore, the objective of this research was to develop untargeted metabolomics using liquid chromatography-high resolution mass spectrometry (LC-HRMS) combined with chemometrics for analysis of pork in beef meatballs for halal authentication. We investigated the use of non-targeted LC-HRMS as a method to detect such food adulteration. As a proof of concept using six technical replicates of pooled samples from beef and pork meat, we could show that metabolomics using LC-HRMS could be used for high-throughput screening of metabolites in meatballs made from beef and pork. Chemometrics of principal component analysis (PCA) was successfully used to differentiate beef meatballs and pork meatball samples. Partial least square-discriminant analysis (PLS-DA) clearly discriminated between halal and non-halal beef meatball samples with 100% accuracy. Orthogonal projection to latent structures-discriminant analysis (OPLS-DA) perfectly discriminated and classified meatballs made from beef, pork, and a mixture of beef-pork with a good level of fitness (R2X = 0.88, R2Y = 0.71) and good predictivity (Q2 = 0.55). Partial least square (PLS) and orthogonal PLS (OPLS) were successfully applied to predict the concentration of pork present in beef meatballs with high accuracy (R2 = 0.99) and high precision. Thirty-five potential metabolite markers were identified through VIP (variable important for projections) analysis. Metabolites of 1-(1Z-hexadecenyl)-sn-glycero-3-phosphocholine, acetyl-l-carnitine, dl-carnitine, anserine, hypoxanthine, linoleic acid, and prolylleucine had important roles for predicting pork in beef meatballs through S-line plot analysis. It can be concluded that a combination of untargeted metabolomics using LC-HRMS and chemometrics is promising to be developed as a standard analytical method for halal authentication of highly processed meat products.