Variation In Metabolites Research Articles

Variation of photosynthesis, secondary metabolites and antioxidant activities in third generation of spaceflight-induced Salvia miltiorrhiza

ObjectiveSpaceflight has long been perceived as an effective way to improve the quantity and quality of plants with wide applications. In order to obtain stable and inheritable descendants of spaceflight-induced Salvia miltiorrhiza lines, we investigated and analyzed four lines m16, m50, m51, m57 (three individuals of each line) and the ground control (three individuals) of the third generation of spaceflight-induced S. miltiorrhiza from primary/secondary metabolism and antioxidative abilities. MethodsA portable photosynthesis system (Li-6400) with red/blue LED light source was used to perform the photosynthetic characteristics to evaluate their primary productivity. The secondary metabolites (phenolic acids, tanshinones, total phenolics and flavonoids) and antioxidant activity of roots were analyzed to assess their quality. ResultsCompared with control, line m16 presented weak photosynthetic ability, but high apparent quantum yield (AQY), higher contents of secondary metabolites, and stronger antioxidative abilities. Line m57 had a strong gas exchange ability, relatively higher secondary metabolites contents, and ascending antioxidative abilities. Lines m50 and m51 were in the middle level of lines m16 and m57. The principal component analysis for all the original data revealed three components including a root-related index, a leaf-related index, and a CO2 response parameter could be used to distinguish spaceflight-induced S. miltiorrhiza lines. ConclusionLine m57 could be an appropriate material for the investigation of targeted breeding towards high production, and line m16 could be used to identify essential genes and unravel sophisticated pathways underlying the secondary metabolisms.

Isomaltooligosaccharides inhibit early colorectal carcinogenesis in a 1,2-dimethylhydrazine-induced rat model.

Colon cancer (CC) is a multistage disease and one of the most common cancers worldwide. Establishing an effective treatment strategies of early colon cancer is of great significance for preventing its development and reducing mortality. The occurrence of colon cancer is closely related to changes in the intestinal flora structure. Therefore, remodelling the intestinal flora structure through prebiotics is a powerful approach for preventing and treating the occurrence and development of colon cancer. Isomaltooligosaccharides (IMOs) are often found in fermented foods and can directly reach the gut for use by microorganisms. In this study, a rat model of early colon cancer (DMH) was established by subcutaneous injection of 1,2-dimethylhydrazine, and the model rats were fed IMOs as a dietary intervention (DI). The untargeted faecal metabolomics, gut metabolome and intestinal function of the model rats were investigated. The results showed that DMH, DI and IMOs alone (IMOs) groups exhibited gut microbial community changes. In the DI group, there was an increased abundance of probiotics (Lactobacillus) and decreased abundance of CC marker bacteria (Fusobacterium). The key variations in the faecal metabolites of the DI group included decreased levels of glucose, bile acids (including deoxycholic acid and chenodeoxycholic acid) and amino acids (including L-glutamic acid and L-alanine). In addition, dietary intake of IMOs attenuated the intestinal inflammatory response, improved the intestinal microecological environment, and slowed the development of DMH-induced early CC in rats. This work provides a theoretical basis and technical support for the clinical prevention or treatment of CC with prebiotics.

Nutritional state variations in a tropical seabird throughout its breeding season

Individual body condition is frequently used to explain differences in foraging and breeding ecology in seabirds. However, little is known about the covariations of body mass with the nutritional state of animals as measured through plasma metabolites and how these different measures vary between and within individuals during breeding. Here, we assessed intra-individual variations of plasma metabolites (triglycerides, cholesterol, protein, and ß-hydroxybutyrate concentrations) and in body mass of Blue-footed boobies (Sula nebouxii) throughout their breeding season 2011–2012 in Isla El Rancho, Mexico. We found breeding-stage and sex-specific variations in individuals’ plasma metabolite concentrations, but these did not mirror variations in body mass. Before egg-laying, females had higher triglycerides, cholesterol, and protein concentrations than males. In contrast, males used their nutritional reserves (higher ß-hydroxybutyrate concentrations) more than females during the breeding season (except for early chick-rearing). At the individual level, males gained weight during the breeding season, whereas females lost weight. We also found that between-individual differences in plasma metabolite concentrations and changes in body mass were not consistent throughout the breeding season, while individual body mass was significantly repeatable. This study contributes to a better understanding of seabird breeding ecology and physiology by showing that sex-specific breeding roles might highly influence the nutritional state. Similar patterns might occur in other seabird species, helping to explain why we can find stage- and sex-specific foraging behaviors even in monomorphic species.

Untargeted serum metabolomics reveals specific metabolite abnormalities in patients with Crohn's disease.

Crohn's disease (CD) is a subtype of inflammatory bowel disease (IBD) characterized by skip intestinal lesions that can occur in any part of the gastrointestinal tract. Currently, the diagnosis of CD is based on clinical history, physical examination and complementary diagnostic tests. It is challenging for physicians to make a definitive diagnosis. This study aimed to analyze the variation in metabolites in CD serum and identify potential predictive biomarkers of CD diagnosis. We collected serum samples from 316 subjects, including patients with CD and healthy controls (HCs). Serum metabolomics was conducted using liquid chromatography coupled to mass spectrometry. Potential biomarkers were screened and evaluated by univariate and multivariate analyses. A panel of two metabolites (deoxycholic acid and palmitic amide) was identified as a specific biomarker of CD. Receiver operating characteristic analysis (ROC) showed that the panel had a sensitivity of 80.25% with a specificity of 95.54% in discriminating CD patients from healthy controls. The biomarkers identified are increased in CD compared with healthy controls. Our approach successfully identified serum biomarkers associated with CD patients. The potential biomarkers indicated that CD metabolic disturbance might be associated with bile acid biosynthesis, fatty acids and energy metabolism.

Dissecting the genetic architecture of glucosinolate compounds for quality improvement in flowering stalk tissues of Brassica napus

Glucosinolates (GSLs) and their hydrolytic products contribute to the quality traits of rapeseed flowering stalk tissues, such as taste, flavor and anticarcinogenic properties (Glucoraphanin). However, little is known about the genetic mechanisms of GSL accumulation in rapeseed flowering stalks. In this study, the variation and genetic architecture of GSL metabolites in flowering stalk tissues were investigated for the first time among a panel of 107 accessions. All GSL compounds exhibited continuous and wide variations in the present population. Progoitrin, glucobrassicanapin and gluconapin were the most abundant GSL compounds. Five quantitative trait loci (QTL) significantly associated with three GSL compounds were identified by genome-wide association study. GRA_C04 was under selected during modern breeding, in which the ratio of lower GSL haplotype (HAP2) in the accessions bred before 1990 (52.56%) was significantly lower than that after 1990 (78.95%). Four candidate genes, BnaA01. SOT16, BnaA06. SOT17, BnaA06. MYB51a, and BnaA06. MYB51b, were identified in the GTL_A01 and 4OH_A06 regions. These findings provide new insights into GSL biosynthesis in flowering stalk tissues and facilitate quality improvement in rapeseed flowering stalks.

Insight into Lotusine and Puerarin in Repairing Alcohol-Induced Metabolic Disorder Based on UPLC-MS/MS.

Alcohol is an essential element in human culture. However, alcoholism has contributed to numerous health issues, including alcoholic fatty liver and sudden death. We found that the alkaloid lotusine possessed hepato- and neuroprotection against alcohol injuries. Lotusine showed comparable protective effects to puerarin, a widely recognized antagonist against alcohol damage. To better understand the metabolic response to alcohol injury and antagonist molecules, we applied sensitive zebrafish and LC-ESI-MS to collect metabolites related to alcohol, puerarin and lotusine exposure. LC-MS identified 119 metabolites with important physiological roles. Differential metabolomic analysis showed that alcohol caused abnormal expression of 82 metabolites (60 up-regulated and 22 down-regulated). These differential metabolites involved 18 metabolic pathways and modules, including apoptosis, necroptosis, nucleotide and fatty acid metabolism. Puerarin reversed seven metabolite variations induced by alcohol, which were related to necroptosis and sphingolipid metabolism. Lotusine was found to repair five metabolites disorders invoked by alcohol, mainly through nucleotide metabolism and glutathione metabolism. In phenotypic bioassay, lotusine showed similar activities to puerarin in alleviating behavioral abnormalities, neuroapoptosis and hepatic lipid accumulation induced by alcohol exposure. Our findings provided a new antagonist, lotusine, for alcohol-induced damage and explored the roles in repairing abnormal metabolism.

Application of 1H-NMR- and LC-MS based Metabolomic analysis for the evaluation of celery preservation methods

In this study, 1H NMR spectroscopy coupled with multivariate analyses was used to evaluate the effectiveness of celery preservation techniques. The extraction parameters were optimised to achieve high recoveries of the majority (9 out of 14) of the quantified metabolites, and to reduce fluctuations in concentrations (within relative standard deviation of 3.5%) and average chemical shifts (0.011 ppm). Subsequently, the optimised method was applied to evaluate metabolite changes caused by different preservation treatments over a 14-day storage period. Nanocellulose treatment (5 g/L) was ascertained to be the most effective from untargeted and targeted principal component analysis (PCA) results, correlating with visual findings, and the significantly reduced metabolite variations (p < 0.05) detected by NMR were identified to be in asparagine, mannitol, malic acid, glucose, and glutamine. LC-MS analysis also revealed significant suppression of phenylpropanoid pathway metabolites (apiin, psoralen, xanthotoxin, and bergapten) by 1.5–25 times. Nanocellulose treatment was an efficient strategy for celery preservation, and NMR- and LC-MS-based metabolomics was an effective approach for establishing reliable metabolomic fingerprints and profiles, enabling the identification and quantitation of metabolite biomarkers for the assessment of storage conditions.

Evaluation of artificial diets based on different legume seeds on the nutritional physiology and digestive function of Helicoverpa armigera (Hübner).

Helicoverpa armigera (Hübner) is considered a serious agricultural pest worldwide. We explored the effects of artificial diets containing ten legumes, including broad beans (Shadan, Feyz, Saraziri, Barekat, and Mahta cultivars), white kidney beans (Dehghan cultivar), red kidney beans (Goli cultivar), common beans (Khomein cultivar), cowpeas (Mashhad and Arabi cultivars) on the feeding responses of H. armigera by quantifying specific primary and secondary metabolites in the studied legumes and determining larval nutritional indices and digestive enzyme activities. The results showed that the highest efficiency of conversion of digested food (ECD) and relative growth rate values (RGR) of whole larval instars were obtained in the Dehghan and Goli cultivars. However, the lowest values of ECD and RGR were observed in the larvae fed on the Khomein and Mahta cultivars. The highest proteolytic and amylolytic activities of larvae were found on the Dehghan and Mashhad cultivars. The highest and lowest values of standardized insect-growth index and index of plant quality were observed in larvae feeding on the Dehghan and Khomein cultivars, respectively. Additionally, significant variations in phytochemical metabolites were recorded among the studied legume cultivars. Significant negative or positive correlations were also found between feeding characteristics and enzymatic activities of H. armigera with the biochemical composition of the studied legumes. The cluster analysis results revealed that artificial diets containing Mahta and Khomein cultivars were unsuitable for H. armigera, and can be used as candidates for integrated pest management programs or for screening insect inhibitors to produce genetically modified pest-resistant plants.

Comparative Metabolomics Analysis Reveals the Variation of Metabolites Associated with Axillary Bud Development in Upland Cotton

Comparative Metabolomics Analysis Reveals the Variation of Metabolites Associated with Axillary Bud Development in Upland Cotton

Proteomic, Metabolomic, and Lipidomic Analyses of Lung Tissue Exposed to Mustard Gas.

Sulfur mustard (HD) poses a serious threat due to its relatively simple production process. Exposure to HD in the short-term causes an inflammatory response, while long-term exposure results in DNA and RNA damage. Respiratory tract tissue models were exposed to relatively low concentrations of HD and collected at 3 and 24 h post exposure. Histology, cytokine ELISAs, and mass spectrometric-based analyses were performed. Histology and ELISA data confirmed previously seen lung damage and inflammatory markers from HD exposure. The multi-omic mass spectrometry data showed variation in proteins and metabolites associated with increased inflammation, as well as DNA and RNA damage. HD exposure causes DNA and RNA damage that results in variation of proteins and metabolites that are associated with transcription, translation and cellular energy.

Using an untargeted metabolomics approach to analyze serum metabolites in COVID-19 patients with nucleic acid turning negative.

Background: The coronavirus disease of 2019 (COVID-19) is a severe public health issue that has infected millions of people. The effective prevention and control of COVID-19 has resulted in a considerable increase in the number of cured cases. However, little research has been done on a complete metabonomic examination of metabolic alterations in COVID-19 patients following treatment. The current project pursues rigorously to characterize the variation of serum metabolites between healthy controls and COVID-19 patients with nucleic acid turning negative via untargeted metabolomics. Methods: The metabolic difference between 20 COVID-19 patients (CT ≥ 35) and 20 healthy controls were investigated utilizing untargeted metabolomics analysis employing High-resolution UHPLC-MS/MS. COVID-19 patients’ fundamental clinical indicators, as well as health controls, were also collected. Results: Out of the 714 metabolites identified, 203 still significantly differed between COVID-19 patients and healthy controls, including multiple amino acids, fatty acids, and glycerophospholipids. The clinical indexes including monocytes, lymphocytes, albumin concentration, total bilirubin and direct bilirubin have also differed between our two groups of participators. Conclusion: Our results clearly showed that in COVID-19 patients with nucleic acid turning negative, their metabolism was still dysregulated in amino acid metabolism and lipid metabolism, which could be the mechanism of long-COVID and calls for specific post-treatment care to help COVID-19 patients recover.

1040 Metabolite Quantification and Entity and Grade Characterisation of Gliomas via Single-Voxel Spectroscopy for Pre-Operative Radiological Diagnosis

Abstract Aim Brain tumours pose a diagnostic challenge, but single-voxel spectroscopy (SVS) provides a non-invasive tool to quantify brain metabolite variations reflective of neoplasia, such as NAA, choline (Cho), creatinine (Cr), lipids (Lip) and lactate (Lac). Inconsistency in spectroscopic research and time-patient pressures emphasises the need for reviewing SVS characterisation of gliomas for peri-operative decision-making. Method A single-centre, retrospective, observational study, including 93 SVS meaningfully interpretable examinations between 2012 and 2018 (PRESS, TE=35ms, 144ms). Spectroscopic diagnoses were compared to histopathologic reports for overall, entity and grade accuracy analysis. Metabolite ratios to Cr were recorded or obtained from peak area integrals. Results High accuracy of lesions discrimination was observed (83–98%). Significant metabolite variation for Cho/Cr (p=0.021) and Cho/NAA (p=0.013) was observed across grades (WHO Grade I-IV). Spectra of astrocytomas and oligodendrogliomas (grade II) were similarly characterised by high Cho, decreased NAA and Cr, and small-to-moderate Lac-Lip and mIns. In contrast, grade III anaplastic oligodendrogliomas (AO) were distinguished from anaplastic astrocytomas (AA) thanks to smaller Lac-Lip and mIns/Cr. Finally, glioblastoma multiforme (GBM, grade IV) displayed high Cho, decreased Cr, the lowest or virtually absent NAA, and the highest Lac-Lip. High-grade spectroscopy, such as enhanced Lac or Lac-Lip, may be an early indicator of grade transformation prior to reflection in histopathology in tumours, which were follow up prospectively. Conclusion This study enhanced spectral characterisation of gliomas and discrimination amongst low- and high-grade gliomas. Whilst these aid pre-operative radiological diagnoses, further research is required to investigate spectroscopy predictive of tumour genetics, now necessary for diagnosis.

Metabolite variation and discrimination of five licorice (Glycyrrhiza) species: HPTLC and NMR explorations

The roots and rhizomes of several Glycyrrhiza species are widely used as sweetening and flavoring agents in food, as well as important ingredients in formulations of traditional medicines. Five Glycyrrhiza species, G. uralensis, G. glabra, G. inflata, G. echinata, and G. lepidota, often share the name "licorice roots" in the botanicals’ marketplace. Unfortunately, misidentification/mislabeling is very common due to their similarities in morpho-anatomical features. Significant metabolite alterations among the different Glycyrrhiza species and their hybrids have been reported, suggesting that the biological activities could vary with the licorice roots or products derived from different species. Development of simple, effective methods for species identification and differentiation is of key importance. In this study, 78 licorice samples were investigated using HPTLC and NMR as analytical tools. Significant metabolite variations were observed between the five species. The species-specific fingerprint patterns for the five Glycyrrhiza species were determined with HPTLC and NMR; then applied to the sample identification and discrimination. The results obtained from these two orthogonal analytical methods agreed with each other. Furthermore, the NMR signals and the species-specific constituents that made significant contributions to the differentiation of the five Glycyrrhiza species were confirmed based on the multivariate analysis of the NMR spectral data. Using the established OPLS-DA models, the classification of hybrids was evaluated and confirmed. The developed methods, particularly the HPTLC method with its simplicity and low cost, could be used as a rapid and reliable approach for the authentication of licorice species and quality control of licorice raw material and products.

1041 Impact of Spectral Contamination and Anatomical Location on Diagnostic Accuracy of Single-Voxel Spectroscopy in Pre-Operative Glioma Characterisation

Abstract Aim Single-voxel spectroscopy (SVS) discriminates amongst brain tumours by measuring metabolite variations reflective of malignant transformation. However, spectral contamination may affect diagnostic accuracy, impacting radiological opinion of regions of interest (ROI) required to determine extent of surgical resection. Method A single-centre, retrospective, observational study, including 93 SVS meaningfully interpretable examinations between 2012 and 2018. Accuracy of lesion classification was assessed by comparison of histological diagnosis as reference standards against discrimination between entities and tumour grades by SVS, with consideration for voxel location, volume, and proximity to the calvaria or ventricular system. Quantification of metabolites was recorded as ratios, and visual inspection, qualitative and descriptive analysis were undertaken. Results SVS demonstrated high entity accuracy (100%) but lower grade accuracy (87%), ranging from periventricular (73%) vs non-periventricular (90%), and subcalvarial (86%) vs subcortical (88%) voxels. CSF contamination caused metabolite dilution and/or enhanced lactate signals from sub-arachnoid or ventricular CSF, especially in areas of increased brain-skull distance; lipid contamination from peri-calvarial fat and subcutaneous tissue resulted in enhanced lipid signals neighbouring areas of increased brain-scalp distance. Misclassified lesions were located regions of increased cranial concavity, reduced cerebral cortical thickness and inappropriate outer volume suppression (OVS). Voxel volume variation had no impact across cerebral coordinates (p=0.605), calvaria proximity (p=0.127) or ventricular proximity (p=0.367). Conclusion Spectral contamination significantly impacted grade accuracy, with metabolite dilution and enhance lactate and lipids signals leading to radiological under- and overestimation of glioma grade, respectively, thus indicating that avoidance of contaminating tissues and OVS are pivotal to adequate spectral quality for diagnostic accuracy.

Analysis of Chlorogenic Acid in Sweet Potato Leaf Extracts.

Sweet potato (Ipomoea batatas L.) is one of the most important food crops worldwide, with leaves of different varieties showing purple, green and yellow, and these leaves provide a dietary source of nutrients and various bioactive compounds. The objective of this study was to identify the active constituents of chlorogenic acids (CGAs) in different methanolic extract of leaves of three varieties of sweet potato (purple CYY 98-59, green Taoyuan 2, and yellow CN 1927-16) using liquid chromatography–tandem mass spectrometry. Genotype-specific metabolite variations were observed; CGAs and three isomeric peaks were detected in sweet potato leaf extracts (SPLEs). Among them, the yellow SPLE contained the highest contents of 3,5-dicaffeoylquinic acid (3,5-di-CQA) and 3,4-dicaffeoylquinic acid (3,4-di-CQA), followed by the green SPLE, whereas the purple SPLE retained lower 3,5-di-CQA content compared to yellow and green SPLEs. All three SPLEs contained lower 4,5-dicaffeoylquinic acid (4,5-di-CQA) and CGA contents compared to 3,5-di-CQA and 3,4-di-CQA, although CGA constituents were not significantly different in genotypes, whereas purple SPLE contained higher 4,5-di-CQA content compared to yellow and green SPLEs. This study indicates that SPLs marketed in Taiwan vary widely in their biological potentials and may impart different health benefits to consumers.

Body Weight and Prandial Variation of Plasma Metabolites in Subjects Undergoing Gastric Band-Induced Weight Loss.

Bariatric procedures are safe and effective treatments for obesity, inducing rapid and sustained loss of excess body weight. Laparoscopic adjustable gastric banding (LAGB) is unique among bariatric interventions in that it is a reversible procedure in which normal gastrointestinal anatomy is maintained. Knowledge regarding how LAGB effects change at the metabolite level is limited. To delineate the impact of LAGB on fasting and postprandial metabolite responses using targeted metabolomics. Individuals undergoing LAGB at NYU Langone Medical Center were recruited for a prospective cohort study. We prospectively analyzed serum samples from 18 subjects at baseline and 2 months after LAGB under fasting conditions and after a 1-hour mixed meal challenge. Plasma samples were analyzed on a reverse-phase liquid chromatography time-of-flight mass spectrometry metabolomics platform. The main outcome measure was their serum metabolite profile. We quantitatively detected over 4,000 metabolites and lipids. Metabolite levels were altered in response to surgical and prandial stimuli, and metabolites within the same biochemical class tended to behave similarly in response to either stimulus. Plasma levels of lipid species and ketone bodies were statistically decreased after surgery whereas amino acid levels were affected more by prandial status than surgical condition. Changes in lipid species and ketone bodies postoperatively suggest improvements in the rate and efficiency of fatty acid oxidation and glucose handling after LAGB. Further investigation is necessary to understand how these findings relate to surgical response, including long term weight maintenance, and obesity-related comorbidities such as dysglycemia and cardiovascular disease.

Plant genetic effects on microbial hubs impact host fitness in repeated field trials.

Although complex interactions between hosts and microbial associates are increasingly well documented, we still know little about how and why hosts shape microbial communities in nature. In addition, host genetic effects on microbial communities vary widely depending on the environment, obscuring conclusions about which microbes are impacted and which plant functions are important. We characterized the leaf microbiota of 200 Arabidopsis thaliana genotypes in eight field experiments and detected consistent host effects on specific, broadly distributed microbial species (operational taxonomic unit [OTUs]). Host genetic effects disproportionately influenced central ecological hubs, with heritability of particular OTUs declining with their distance from the nearest hub within the microbial network. These host effects could reflect either OTUs preferentially associating with specific genotypes or differential microbial success within them. Host genetics associated with microbial hubs explained over 10% of the variation in lifetime seed production among host genotypes across sites and years. We successfully cultured one of these microbial hubs and demonstrated its growth-promoting effects on plants in sterile conditions. Finally, genome-wide association mapping identified many putatively causal genes with small effects on the relative abundance of microbial hubs across sites and years, and these genes were enriched for those involved in the synthesis of specialized metabolites, auxins, and the immune system. Using untargeted metabolomics, we corroborate the consistent association between variation in specialized metabolites and microbial hubs across field sites. Together, our results reveal that host genetic variation impacts the microbial communities in consistent ways across environments and that these effects contribute to fitness variation among host genotypes.

An LC–MS-based metabolomic approach provides insights into the metabolite profiles of Ginkgo biloba L. at different developmental stages and in various organs

Ginkgo (Ginkgo biloba L.) is a tree valued for the high medicinal and nutritional value of its leaves and seeds. However, the metabolite profiles of ginkgo leaves and seeds and their changes during development have not been comprehensively analyzed, which hinders improvements in the utilization of ginkgo. A comprehensive and systematic untargeted LC–MS metabolomics analysis of the metabolites in ginkgo leaves (male and female) and seeds at two developmental stages identified 8146 known metabolites, which mainly included lipids and lipid-like molecules, phenylpropanoids and polyketides, organoheterocyclic compounds, organic acids and derivatives, organic oxygen compounds, and benzenoids. Some of the identified metabolites have known healthcare and food value, and some of the others are newly discovered metabolites with potential for new drug development. The small number of differential Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways between leaves of male and female gingko trees indicated that the developmental stage affected the metabolic pathways more significantly than sex. Among the flavonoid constituents of ginkgo, 653 flavonoid metabolites were identified, and these included some new flavonoid components, which confirmed that the developmental stage is a critical factor in secondary metabolite variations. This study illuminated the metabolites and medicinal and edible values of ginkgo leaves and seeds at different developmental stages and thus supports further effective utilization of ginkgo leaves and seeds.

Effect of exogenous jasmonic acid on physiology and steroidal saponin accumulation in Dioscorea zingiberensis

Dioscorea zingiberensis is a valuable medicinal herb rich in steroidal saponins. To reveal the role of jasmonic acid (JA) on physiology and steroidal saponins accumulation, D. zingiberensis were treated with different concentrations of JA. The antioxidant capacity, photosynthetic parameters, fatty acids and metabolites related to steroidal saponins biosynthesis (phytosterols, diosgenin and steroidal saponins) were examined under JA treatment. The results demonstrated that JA treatment caused a great reduction in MDA, stomatal width, photosynthetic rate and photosynthetic pigment, induced a considerable increase in proline, soluble sugar, soluble protein and antioxidant enzymes (CAT, POD and SOD), and leaded to a significant up-regulation in the expression of genes related to antioxidant system and chlorophyll degradation. Specialized metabolites displayed various changes under different concentrations of JA. The majority of fatty acids exhibited negative responses to JA treatment in leaf and rhizome. In leaf, JA treatment enhanced the accumulation of phytosterols and diosgenin, but decreased the accumulation of steroidal saponins. However, steroidal saponins were mainly accumulated in rhizome and were highly increased by JA treatment. Redundancy analysis illustrated that fatty acids were strongly associated with metabolites related to steroidal saponins. Among all fatty acids, C16:0, C18:1, C18:3, C22:0 and C24:0 contributed most to the variation in metabolites related to steroidal saponin biosynthesis. Overall, JA treatment leaded to an increase in steroidal saponins, but an inhibition of plant growth. Thus, the negative effects of JA application on plant physiology should be carefully assessed before being utilized to increase the production of steroidal saponins in D. zingiberensis.

Sources of Variation in Food-Related Metabolites during Pregnancy.

The extent to which variation in food-related metabolites are attributable to non-dietary factors remains unclear, which may explain inconsistent food-metabolite associations observed in population studies. This study examined the association between non-dietary factors and the serum concentrations of food-related biomarkers and quantified the amount of variability in metabolite concentrations explained by non-dietary factors. Pregnant women (n = 600) from two Canadian birth cohorts completed a validated semi-quantitative food frequency questionnaire, and serum metabolites were measured by multisegment injection-capillary electrophoresis-mass spectrometry. Hierarchical linear modelling and principal component partial R-square (PC-PR2) were used for data analysis. For proline betaine and DHA (mainly exogenous), citrus foods and fish/fish oil intake, respectively, explained the highest proportion of variability relative to non-dietary factors. The unique contribution of dietary factors was similar (15:0, 17:0, hippuric acid, TMAO) or lower (14:0, tryptophan betaine, 3-methylhistidine, carnitine) compared to non-dietary factors (i.e., ethnicity, maternal age, gestational age, pre-pregnancy BMI, physical activity, and smoking) for metabolites that can either be produced endogenously, biotransformed by gut microbiota, and/or derived from multiple food sources. The results emphasize the importance of adjusting for non-dietary factors in future analyses to improve the accuracy and precision of the measures of food intake and their associations with health and disease.