Non-targeted Metabolite Research Articles

Investigation of Xenobiotics Metabolism In Salix alba Leaves via Mass Spectrometry Imaging.

The method presented uses mass spectrometry imaging (MSI) to establish the metabolic profile of S. alba leaves when exposed to xenobiotics. Using a non-targeted approach, plant metabolites and xenobiotics of interest are identified and localized in plant tissues to uncoverspecific distribution patterns. Then, in silico prediction of potential metabolites (i.e., catabolites and conjugates) from the identified xenobiotics is performed. When a xenobiotic metabolite is located in the tissue, the type of enzyme involved in its alteration by the plant is recorded. These results were used to describe different types of biological reactions occurring in S. alba leaves in response to xenobiotic accumulation in the leaves. The metabolites were predicted in two generations, allowing the documentation of successive biological reactions to transform xenobiotics in the leaf tissues.

The protective effect of losartan against sorafenib induced cardiotoxicity: Ex-vivo isolated heart and metabolites profiling studies in rat

Sorafenib, a tyrosine kinase inhibitor that is used in the treatment of hepatocellular and renal cell carcinoma, was reported to induce cardiotoxicity. This study aimed to investigate the potential cardioprotective effect of losartan against sorafenib-induced cardiotoxicity in rat. Sorafenib significantly reduced the left ventricular pressure, heart rate dp/dt max & dp/dt min (indexes of myocardial contractility and relaxation; respectively), and prolonged both the systolic and diastolic periods. Coadminstration of losartan significantly reversed the effects of sorafenib on heart rate, dp/dt max and dp/dt min. In addition, there was a tendency for losartan to reverse sorafenib reduction in left ventricular pressure and perfusion pressure but it did not reach statistical significance. A GC-MS non-targeted based metabolites profiling of rat plasma revealed elevated metaboites, including urea and fatty acids levels, associated with sorafenib induced cardiotoxicity. However, only glycine and lactic acid were statistically significant. Interestingly, losartan co-administration with sorafenib restored these changes, and resulted in a significantly reduced glycine, urea and some fatty acids levels namely; Cis-vaccenic acid, oleic acid, stearic acid and undecanoic acid. In addition, based on histology results, losartan coadminitration almost obviated sorafenib-induced changes in cardiac tissues. The study suggests that losartan has the potential to exert a protective effect against sorafenib-induced cardiotoxicity.

Non-Targeted Metabolite Profiles and Sensory Properties Elucidate Commonalities and Differences of Wines Made with the Same Variety but Different Cultivar Clones.

Grapes, one of the oldest agricultural crops, are cultivated to produce table fruits, dried fruits, juice, and wine. Grapevine variety is composed of clones that share common morphological traits. However, they can differ in minor genetic mutations which often result in not only notorious morphological changes but also in other non-visible sensorial distinctive attributes. In the present work, we identified three Vitis vinifera cv. Pinot noir clones grown under identical field conditions that showed different grape cluster types. Here, sensorial analysis together with non-targeted metabolite profiles by Ultra High performance Liquid Chromatography (UPLC) couples to Ultra High Resolution Mass Spectrometry (FT-ICR-MS) of wines elaborated from the three different grape cluster types was studied with the aim of (i) finding sensorial differences among these three types of wines, and, if there were, (ii) determining the molecular features (metabolites) associated with these sensorial attributes by a multivariate statistical approach.

Dairy Intake in 2 American Adult Cohorts Associates with Novel and Known Targeted and Nontargeted Circulating Metabolites

The role of dairy in health can be elucidated by investigating circulating metabolites associated with intake. We sought to identify metabolites associated with quantity and type of dairy intake in the Framingham Heart Study Offspring and Third Generation (Gen3) cohorts. Dairy intake (total dairy, milk, cheese, yogurt, and cream/butter) was analyzed in relation to targeted (Offspring, n=2205, 55.1±9.8y, 52% female, 217 signals; Gen3, n=866, 40.5±8.8y, 54.9% female, 79 signals) and nontargeted metabolites (Gen3, ∼7031 signals) in a 2-step analysis including orthogonal projections to latent structures with discriminant analysis (OPLS-DA) in discovery subsets to identify metabolites distinguishing between high and low intake; and linear regression in confirmation subsets to assess putative associations, subsequently tested in the total samples. Previously reported associations were also investigated. OPLS-DA in the Offspring targeted discovery subset resulted in a variable importance in projection (VIP) >1 of 65, 60, 58, 66, and 60 metabolites for total dairy, milk, cream/butter, cheese, and yogurt, respectively, of which 5, 3, 1, 6, and 4 metabolites, respectively, remained after confirmation. In the Gen3 targeted discovery subset, OPLS-DA resulted in a VIP >1 of 17, 15, 13, 7, and 6 metabolites for total dairy, milk, cream/butter, cheese, and yogurt, respectively. In the Gen3 nontargeted discovery subset, OPLS-DA resulted in a VIP >2 of 203, 503, 78, 186, and 206 metabolites, respectively. Combining targeted and nontargeted results in Gen3, significant associations of 7 (6 unannotated), 2, 12 (11 unannotated), 0, and 61 (all unannotated) metabolites, respectively, remained. Candidate identities of unannotated signals included fatty acids and food flavorings. Results supported relations previously reported for C14:0 sphingomyelin, and marginal associations for deoxycholates. Dairy in 2 American adult cohorts associated with numerous circulating metabolites. Reports about diet-metabolite relations and confirmation of previous findings might be limited by specificity of dietary intake and breadth of measured metabolites.

Metabolite profiling of whiteleg shrimp Litopenaeus vannamei from super-intensive culture in closed aquaculture systems: a recirculating aquaculture system and a hybrid zero water discharge-recirculating aquaculture system.

The production of the whiteleg shrimp Litopenaeus vannamei now accounts for approximately 75% of the total shrimp production in Indonesia. The techniques used to produce whiteleg shrimp in Indonesia are still dominated by conventional rearing strategies using open-pond systems, which often contribute to unpredictable culture performance and weak sustainability. Alternative production strategies of closed aquaculture systems, including the recirculating aquaculture system (RAS) and hybrid zero water discharge-recirculating aquaculture system (hybrid system), have been developed and implemented for higher productivity, stability and sustainability of whiteleg shrimp grow-out production in Indonesia. Despite the positive aspects of the application of closed aquaculture systems in shrimp aquaculture, the differences in the characteristics of shrimp grown in closed RAS and hybrid systems compared to open-pond systems remain unclear. This study aims to investigate the differences in the metabolite profiles of shrimp grown in intensive closed aquaculture systems, including an RAS and hybrid system, compared to those of shrimp grown in a semi-intensive, open, earthen pond system by means of non-targeted GC-MS metabolite profiling. Shrimp cultured in the closed systems (RAS and hybrid system) and an open system (pond) were harvested and subjected to GC-MS non-targeted metabolomics analysis. A total of 112 metabolites were annotated from shrimp samples and subjected to principal component analysis (PCA). The metabolites annotated from GC-MS mainly included organic compounds, proteinogenic and non-proteinogenic amino acids, sugars, nucleosides and fatty acids. The results of principal component analysis showed several metabolites with high variable importance in projection (VIP) scores, including shikimic acid, β-alanine, uric acid, hypoxanthine, inosine, homocysteine, methionine, phenylalanine, tryptophan and lysine, as the main metabolites differentiating the shrimp grown in the three production systems. Our findings showed that shrimp cultured in different aquaculture systems exhibited distinct metabolite profiles, and the metabolites showing high VIP scores, including shikimic acid, β-alanine, uric acid, hypoxanthine, inosine, homocysteine, methionine, phenylalanine, tryptophan and lysine, may serve as candidate markers to indicate the differences in shrimp from different production systems.

Non-targeted urinary metabolomics in pregnancy and associations with fetal growth restriction

Our objective was to identify metabolites associated with fetal growth restriction (FGR) by examining early and late pregnancy differences in non-targeted urinary metabolites among FGR cases and non-FGR controls. An exploratory case-control study within LIFECODES birth cohort was performed. FGR cases (N = 30), defined as birthweight below the 10th percentile, were matched with controls (N = 30) based on maternal age, race, pre-pregnancy body mass index, and gestational age at delivery. Gas chromatography/electron-ionization mass spectrometry was performed on urine samples collected at 10 and 26 weeks of gestation. Differences in urinary metabolite levels in cases and controls at each time point and between the two time points were calculated and then changes compared across pregnancy. 137 unique urinary metabolites were annotated, and several identified that were higher in cases compared to controls. For example, urinary concentrations of benzoic acid were higher in cases compared to controls at both study visits (3.01-fold higher in cases at visit 1, p < 0.01; 3.10-fold higher in cases at visit 3, p = 0.05). However, these findings from our exploratory analysis were not robust to false-discovery-rate adjustment. In conclusion, using a high-resolution, non-targeted approach, we found specific urinary organic acids differed over pregnancy by FGR case status.

Discrimination of Adzuki Bean (Vigna angularis) Geographical Origin by Targeted and Non-Targeted Metabolite Profiling with Gas Chromatography Time-of-Flight Mass Spectrometry.

As international food trade increases, consumers are becoming increasingly interested in food safety and authenticity, which are linked to geographical origin. Adzuki beans (Vigna angularis) are cultivated worldwide, but there are no tools for accurately discriminating their geographical origin. Thus, our study aims to develop a method for discriminating the geographical origin of adzuki beans through targeted and non-targeted metabolite profiling with gas chromatography time-of-flight mass spectrometry combined with multivariate analysis. Orthogonal partial least squares discriminant analysis showed clear discrimination between adzuki beans cultivated in Korea and China. Non-targeted metabolite profiling showed better separation than targeted profiling. Furthermore, citric acid and malic acid were the most notable metabolites for discriminating adzuki beans cultivated in Korea and China. The geographical discrimination method combining non-targeted metabolite profiling and pareto-scaling showed excellent predictability (Q2 = 0.812). Therefore, it is a suitable prediction tool for the discrimination of geographical origin and is expected to be applicable to the geographical authentication of adzuki beans.

Metabolomics should be deployed in the identification and characterization of gene-edited crops.

Gene-editing techniques are currently revolutionizing biology, allowing far greater precision than previous mutagenic and transgenic approaches. They are becoming applicable to a wide range of plant species and biological processes. Gene editing can rapidly improve a range of crop traits, including disease resistance, abiotic stress tolerance, yield, nutritional quality and additional consumer traits. Unlike transgenic approaches, however, it is not facile to forensically detect gene-editing events at the molecular level, as no foreign DNA exists in the elite line. These limitations in molecular detection approaches are likely to focus more attention on the products generated from the technology than on the process in itself. Rapid advances in sequencing and genome assembly increasingly facilitate genome sequencing as a means of characterizing new varieties generated by gene-editing techniques. Nevertheless, subtle edits such as single base changes or small deletions may be difficult to distinguish from normal variation within a genotype. Given these emerging scenarios, downstream 'omics' technologies reflective of edited affects, such as metabolomics, need to be used in a more prominent manner to fully assess compositional changes in novel foodstuffs. To achieve this goal, metabolomics or 'non-targeted metabolite analysis' needs to make significant advances to deliver greater representation across the metabolome. With the emergence of new edited crop varieties, we advocate: (i) concerted efforts in the advancement of 'omics' technologies, such as metabolomics, and (ii) an effort to redress the use of the technology in the regulatory assessment for metabolically engineered biotech crops.

Evaluation of the contribution of trichomes to metabolite compositions of tea (Camellia sinensis) leaves and their products

Tea is processed from the postharvest tender leaves from tea plants (Camellia sinensis). Leaves containing more trichomes are considered to produce higher quality tea, but no evidence is available to support this opinion. In the present study, morphological and metabolic analyses of trichomes showed that tea trichomes were unicellular, simple, and rich in many water-soluble metabolites, including free catechins, caffeine, and amino acids. Furthermore, the analysis of known quality-related and non-targeted metabolites in fresh tea leaves and tea products showed that the presence of trichomes did not change the overall metabolite profiles of fresh tea leaves, white tea, and black tea, but might affect the non-targeted metabolites in green tea. Overall, more trichomes might only indicate higher quality tea leaves, while metabolites in the leaf might be the key factor. This study provides the first determination of the contribution of trichomes to tea quality.

Aquatic macroinvertebrates under stress: Bioaccumulation of emerging contaminants and metabolomics implications

The current knowledge on bioaccumulation of emerging contaminants (ECs) in aquatic invertebrates exposed to the realistic environmental concentrations is limited. Even less is known about the effects of chemical pollution exposure on the metabolome of aquatic invertebrates. We conducted an in situ translocation experiment with passive filter-feeding caddisfly larvae (Hydropsyche sp.) in an effluent-influenced river in order to i) unravel the bioaccumulation (and recovery) dynamics of ECs in aquatic invertebrates, and ii) test whether exposure to environmentally realistic concentrations of ECs will translate into metabolic profile changes in the insects. The experiment was carried out at two sites, upstream and downstream of the discharge of an urban wastewater treatment plant effluent. The translocated animals were collected at 2-week intervals for 46 days. Both pharmaceuticals and endocrine disrupting compounds (EDCs) were detected in water (62 and 7 compounds, respectively), whereas in Hydropsyche tissues 5 EDCs accumulated. Overall, specimens from the upstream site translocated to the impacted site reached higher ECs concentrations in their tissues, as a reflection of the contaminants’ water concentrations. However, bioaccumulation was a temporary process susceptible to change under lower contaminant concentrations. Non-targeted metabolite profiling detected fine metabolic changes in translocated Hydropsyche larvae. Both translocations equally induced stress, but it was higher in animals translocated to the impacted site.

Causal effects of serum metabolites on amyotrophic lateral sclerosis: A Mendelian randomization study

Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder that is affected by both genetic and environmental factors. Nowadays, OMIC technologies, such as genomics and metabolomics, are providing a systematic readout of genetic structures and physiological states for understanding human diseases. However, the comprehensive analysis of cross-omics is often lacking. Here, we conducted a Mendelian randomization analysis to provide a comprehensive analysis of metabolomics and genomics to estimate the causal relationships between non-targeted human serum metabolites and the development of ALS. Using genetic variants as predictors, our study detected 18 metabolites that might have causal effects on the development of ALS, including a group of gamma-glutamyl amino acids. Our findings suggested that glutathione metabolism dysfunction might be involved in the pathogenesis of ALS. Furthermore, our study provides a novel method to understand the pathogenesis of human diseases and develop therapeutic strategies for diseases by combining metabolomics with genomics.

Random forest-based imputation outperforms other methods for imputing LC-MS metabolomics data: a comparative study

BackgroundLC-MS technology makes it possible to measure the relative abundance of numerous molecular features of a sample in single analysis. However, especially non-targeted metabolite profiling approaches generate vast arrays of data that are prone to aberrations such as missing values. No matter the reason for the missing values in the data, coherent and complete data matrix is always a pre-requisite for accurate and reliable statistical analysis. Therefore, there is a need for proper imputation strategies that account for the missingness and reduce the bias in the statistical analysis.ResultsHere we present our results after evaluating nine imputation methods in four different percentages of missing values of different origin. The performance of each imputation method was analyzed by Normalized Root Mean Squared Error (NRMSE). We demonstrated that random forest (RF) had the lowest NRMSE in the estimation of missing values for Missing at Random (MAR) and Missing Completely at Random (MCAR). In case of absent values due to Missing Not at Random (MNAR), the left truncated data was best imputed with minimum value imputation. We also tested the different imputation methods for datasets containing missing data of various origin, and RF was the most accurate method in all cases. The results were obtained by repeating the evaluation process 100 times with the use of metabolomics datasets where the missing values were introduced to represent absent data of different origin.ConclusionType and rate of missingness affects the performance and suitability of imputation methods. RF-based imputation method performs best in most of the tested scenarios, including combinations of different types and rates of missingness. Therefore, we recommend using random forest-based imputation for imputing missing metabolomics data, and especially in situations where the types of missingness are not known in advance.

Metabolic Trajectories Following Contrasting Prudent and Western Diets from Food Provisions: Identifying Robust Biomarkers of Short-Term Changes in Habitual Diet.

A large body of evidence has linked unhealthy eating patterns with an alarming increase in obesity and chronic disease worldwide. However, existing methods of assessing dietary intake in nutritional epidemiology rely on food frequency questionnaires or dietary records that are prone to bias and selective reporting. Herein, metabolic phenotyping was performed on 42 healthy participants from the Diet and Gene Intervention (DIGEST) pilot study, a parallel two-arm randomized clinical trial that provided complete diets to all participants. Matching single-spot urine and fasting plasma specimens were collected at baseline, and then following two weeks of either a Prudent or Western diet with a weight-maintaining menu plan designed by a dietician. Targeted and nontargeted metabolite profiling was conducted using three complementary analytical platforms, where 80 plasma metabolites and 84 creatinine-normalized urinary metabolites were reliably measured (CV < 30%) in the majority of participants (>75%) after implementing a rigorous data workflow for metabolite authentication with stringent quality control. We classified a panel of metabolites with distinctive trajectories following two weeks of food provisions when using complementary univariate and multivariate statistical models. Unknown metabolites associated with contrasting dietary patterns were identified with high-resolution MS/MS, as well as co-elution after spiking with authentic standards if available. Overall, 3-methylhistidine and proline betaine concentrations increased in both plasma and urine samples after participants were assigned a Prudent diet (q < 0.05) with a corresponding decrease in the Western diet group. Similarly, creatinine-normalized urinary imidazole propionate, hydroxypipecolic acid, dihydroxybenzoic acid, and enterolactone glucuronide, as well as plasma ketoleucine and ketovaline increased with a Prudent diet (p < 0.05) after adjustments for age, sex, and BMI. In contrast, plasma myristic acid, linoelaidic acid, linoleic acid, α-linoleic acid, pentadecanoic acid, alanine, proline, carnitine, and deoxycarnitine, as well as urinary acesulfame K increased among participants following a Western diet. Most metabolites were also correlated (r > ± 0.30, p < 0.05) to changes in the average intake of specific nutrients from self-reported diet records reflecting good adherence to assigned food provisions. Our study revealed robust biomarkers sensitive to short-term changes in habitual diet, which is needed for accurate monitoring of healthy eating patterns in free-living populations, and evidence-based public health policies for chronic disease prevention.

Effects of Environmental Pollution on Changes in Blood Biochemical Parameters

&#x0D; &#x0D; &#x0D; &#x0D; &#x0D; Abstract &#x0D; &#x0D; A metabolic marker in the blood of individuals exposed to air pollution is plausible as some environmental pollutants have been observed to enter the blood stream directly from lungs. The main objective of the current review is to find out changes in biochemical parameters of human blood as a result of environmental pollution. Spatio temporal models involving measurement of Air Quality Index were accommodating to find out the results of development of an aggregate air quality index. Results depicted that mean values of RBC and hemoglobin concentration in blood of exposed children were significantly different from those of non-exposed group. Air pollution may adversely affect children's erythrocytes, particularly PM10 have a significant negative relationship with hemoglobin and RBC numbers while a positive significant relationship with WBC and platelet count. Higher concentrations of air pollutants exposed to pregnant women significantly higher the incidence of pregnancy anemia’s. A strong dose-response relationship was confirmed for both contaminants. It is difficult to establish a causal relationship between specific environmental exposures and complicated multifactorial health outcomes, the application of non-targeted metabolite profiling to assess the effect of air pollution on blood metabolite. Therefore in the present study the main emphasis has been made on the effects of various polluted environment contaminants, their biochemical effects on the composition of blood and various associated disease.&#x0D; &#x0D; &#x0D; &#x0D; &#x0D; &#x0D; &#x0D; &#x0D; &#x0D; &#x0D;

Comparative Evaluation of Six Traditional Fermented Soybean Products in East Asia: A Metabolomics Approach.

Many ethnic fermented soybean products (FSPs) have long been consumed as seasoning and protein sources in East Asia. To evaluate the quality of various FSPs in East Asia, non-targeted metabolite profiling with multivariate analysis of six traditional FSPs (Natto; NT, Cheonggukjang; CG, Doenjang; DJ, Miso; MS, Doubanjiang; DB, Tianmianjiang; TM) was performed. Six FSPs could be clearly distinguished by principle component analysis (PCA) and partial least square-discriminant analysis (PLS-DA). Amino acid contents were relatively higher in NT and CG, sugar and sugar alcohol contents were relatively higher in MS and TM, isoflavone glycoside contents were relatively highest in CG, isoflavone aglycon contents were the highest in DJ, and soyasaponin contents were the highest in CG. Antioxidant activity and physicochemical properties were determined to examine the relationships between the FSPs and their antioxidant activities. We observed a negative correlation between isoflavone aglycon contents and 2,2’-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) activity. Furthermore, the order of ABTS activity of FSPs has a positive correlation with the order of soybean content in the six FSPs. Herein it was found that primary metabolites were affected by the main ingredients and secondary metabolites were most influenced by the fermentation time, and that soybean content contributed more to antioxidant activity than fermentation time.

Metabolic Perturbations from Step Reduction in Older Persons at Risk for Sarcopenia: Plasma Biomarkers of Abrupt Changes in Physical Activity

Sarcopenia is the age-related loss of skeletal muscle mass, strength and function, which may be accelerated during periods of physical inactivity. Declines in skeletal muscle and functionality not only impacts mobility but also increases chronic disease risk, such as type 2 diabetes. The aim of this study was to measure adaptive metabolic responses to acute changes in habitual activity in a cohort of overweight, pre-diabetic older adults (age = 69 ± 4 years; BMI = 27 ± 4 kg/m2, n = 17) when using non-targeted metabolite profiling by multisegment injection-capillary electrophoresis-mass spectrometry. Participants completed two weeks of step reduction (<1000 steps/day) followed by a two week recovery period, where fasting plasma samples were collected at three time intervals at baseline, after step reduction and following recovery. Two weeks of step reduction elicited increases in circulatory metabolites associated with a decline in muscle energy metabolism and protein degradation, including glutamine, carnitine and creatine (q < 0.05; effect size > 0.30), as well as methionine and deoxycarnitine (p < 0.05; effect size ≈ 0.20) as compared to baseline. Similarly, decreases in uremic toxins in plasma that promote muscle inflammation, indoxyl sulfate and hippuric acid, as well as oxoproline, a precursor used for intramuscular glutathione recycling, were also associated with physical inactivity (p < 0.05; effect size > 0.20). Our results indicate that older persons are susceptible to metabolic perturbations due to short-term step reduction that were not fully reversible with resumption of normal ambulatory activity over the same time period. These plasma biomarkers may enable early detection of inactivity-induced metabolic dysregulation in older persons at risk for sarcopenia not readily measured by current imaging techniques or muscle function tests, which is required for the design of therapeutic interventions to counter these deleterious changes in support of healthy ageing.

1389-P: Nontargeted Metabolomics Reveal Novel Metabolites in Small- and Large-for-Gestational-Age Infants and Their Mothers Associated with Glucose and Lipid Metabolism

Small-for-gestational-age (SGA) or large-for-gestational-age (LGA) infants have increased risk of type 2 diabetes mellitus in later life. The intrauterine environment provides a foundation on which chronic disease develop in adulthood, however, the molecular mechanisms still remained unsolved. We recruited 135 pregnant and their infants, and then selected 67 pairs for plasma metabolomics analysis according to the birthweight of newborns, SGA (&amp;lt;10th percentile, n=16), AGA (∼50th percentile, n=28) and LGA (&amp;gt;90th percentile, n=23). Non-targeted metabolomics were performed by UPLC-Q-TOF-MS, and the metabolites were screened by P value, fold change and VIP value in OPLS-DA model. We found a group of metabolites over-/low-expressed in both SGA and LGA groups when compared to AGA group. Cholesteryl acetate (CE) was negatively related to SGA and LGA (logistic regression, AGA=0, SGA and LGA=1) in both mothers (adj. r=-1.50, adj. p=0.03) and infants (adj. r=-1.22, adj. p=0.03). Citicoline was positively related to SGA and LGA (maternal adj. r=4.71, adj. p=0.18; fetal adj. r=0.97, adj. p=0.06). CE and citicoline were involved in glycerophospholipid metabolism (EHMN database). Dihydroxyfumaric acid (DA) reported as insulin inhibitor was over-expressed in SGA and LGA (maternal adj. r=0.72, adj. p=0.09; fetal adj. r=0.91, adj. p=0.09). Moreover, the fetal citicoline (r=-0.294, p=0.082) and CE (r=0.369, p=0.027) was associated with cord blood glucose. Fetal DA was negatively associated with HDL (r=-0.416, p=0.012) and total cholesterol (r=-0.345, p=0.039). In conclusion, the abnormal expression of CE, citicoline and DA in SGA and LGA mothers might cause the same abnormality in fetus, and were involved in glycerophospholipid metabolism and insulin inhibition. These new findings indicated that a group of metabolites which can be passed from mother to offspring might affect the metabolic syndrome in both SGA and LGA infants. Disclosure J. Liu: None. X. Zhai: None. X. Xiao: None. M. Yu: None. N. Zhao: None. Z. Li: None. J. Fu: None. M. Deng: None. L. Zhou: None. Funding National Natural Science Foundation of China (81870579); Fundamental Research Funds for the Central Universities (3332018015)

Metabolomics reveals elevated urinary excretion of collagen degradation and epithelial cell turnover products in irritable bowel syndrome patients.

Irritable bowel syndrome (IBS), the most commonly diagnosed functional gastrointestinal (GI) disorder in developed countries, is characterized by chronic abdominal pain, and altered bowel habits. Accurate and timely diagnosis is challenging as it relies on symptoms and an evolving set of exclusion criteria to distinguish it from other related GI disorders reflecting a complex etiology that remains poorly understood. Herein, nontargeted metabolite profiling of repeat urine specimens collected from a cohort of IBS patients (n = 42) was compared to healthy controls (n = 20) to gain insights into the underlying pathophysiology. An integrated data workflow for characterization of the urine metabolome with stringent quality control was developed to authenticate reliably measured (CV < 30%) and frequently detected (> 75%) metabolites using multisegment injection-capillary electrophoresis-mass spectrometry. Complementary statistical methods were then used to rank differentially excreted urinary metabolites after normalization to osmolality that were subsequently identified by high resolution tandem mass spectrometry and their electrophoretic migration behavior. Our work revealed ten consistently elevated urinary metabolites in repeat samples collected from IBS patients at two different time points (q < 0.05 after age and Benjamini-Hochberg/FDR adjustment), which were associated with greater collagen degradation and intestinal mucosal turn-over processes likely due to low-grade inflammation. IBS-specific metabolites identified in urine included a series of hydroxylysine metabolites (O-glycosylgalactosyl-hydroxylysine, O-galactosyl-hydroxylysine, lysine), mannopyranosy-L-tryptophan, imidazole propionate, glutamine, serine, ornithine, dimethylglycine and dimethylguanosine. A major limitation in this retrospective case-control study was significant co-morbidity of IBS patients with other illnesses, including depression and prescribed medications as compared to healthy controls. This work provides new mechanistic insights into the pathophysiology of IBS while also offering a convenient way to monitor patient disease progression and treatment responses to therapy based on a panel of urinary metabolites that avoids invasive blood sampling, colonoscopy and/or tissue biopsies.

GC–MS-Based Nontargeted and Targeted Metabolic Profiling Identifies Changes in the Lentinula edodes Mycelial Metabolome under High-Temperature Stress

To clarify the physiological mechanism of the Lentinula edodes (L. edodes) response to high-temperature stress, two strains of L. edodes with different tolerances were tested at different durations of high temperature, and the results showed that there were significant changes in their phenotypes and physiology. To further explore the response mechanism, we established a targeted GC–MS-based metabolomics workflow comprising a standardized experimental setup for growth, treatment and sampling of L. edodes mycelia, and subsequent GC–MS analysis followed by data processing and evaluation of quality control (QC) measures using tailored statistical and bioinformatic tools. This study identified changes in the L. edodes mycelial metabolome following different time treatments at high temperature based on nontargeted metabolites with GC-MS and further adopted targeted metabolomics to verify the results of the analysis. After multiple statistical analyses were carried out using SIMCA software, 74 and 108 differential metabolites were obtained, respectively. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis showed that the metabolic pathways with significant changes included those related to the following: amino acid metabolism, the glycolysis pathway, the tricarboxylic acid (TCA) cycle, and sugar metabolism. Most amino acids and carbohydrates enriched in these metabolic pathways were upregulated in strain 18, downregulated in strain 18N44, or the synthesis in strain 18 was higher than that in strain 18N44. This result was consistent with the physiological phenotypic characteristics of the two strains under high-temperature stress and revealed the reason why strain 18N44 was more heat-sensitive. At the same time, under high temperature, the decrease of intermediate products in glycolysis and the TCA cycle resulted in carbon starvation and insufficient energy metabolism, thus inhibiting the growth of L. edodes. In addition, the results also showed that the metabolites produced by different L. edodes strains under high-temperature stress were basically the same. However, different strains had species specificity, so the changes in the content of metabolites involved in the response to high-temperature stress were different. This provides a theoretical basis for further understanding the mechanism of the L. edodes response to high temperature and can be used to establish an evaluation system of high-temperature-resistant strains and lay the foundation for molecular breeding of new L. edodes strains resistant to high temperature.

OR07-1 Cord Blood Metabolomics: Association with Newborn Anthropometrics and C-Peptide across Ancestries

Newborn adiposity is associated with a higher risk of childhood obesity and earlier onset of co-morbid metabolic diseases such as type 2 diabetes mellitus. The cord blood metabolome is one early life marker that provides mechanistic insight into fetal fat deposition, insulin sensitivity, and future obesity and metabolic disease risk and integrates in-utero genetic, nutritional and metabolic cues that contribute to newborn phenotype. We hypothesize that a unique cord blood metabolomic signature associated with newborn adiposity or hyperinsulinemia might emerge as a predictive tool to help identify at-risk children early in life, before disease develops. To evaluate this hypothesis, we performed a cross-sectional, observational study of 1600 newborns who participated in the Hyperglycemia and Adverse Pregnancy Outcome Study. Targeted and nontargeted metabolomics assays were performed on cord blood of newborns across four ancestry groups (Afro-Caribbean, Northern European, Thai, and Mexican American). Newborn birth weight and sum of skinfolds were measured by trained personnel and cord blood was additionally assayed for C-peptide. Associations of cord blood metabolites with newborn phenotype were investigated using 1) per-metabolite analyses within and across ancestries and 2) network analyses to identify interconnected metabolites associated with phenotypes. Several metabolites, including branched-chain amino acids, medium- and long-chain acylcarnitines, nonesterified fatty acids, and triglycerides were significantly negatively associated with cord C-peptide but positively associated with birth weight and/or sum of skinfolds. 1,5-Anhydroglucitol was significantly associated with birth weight and sum of skinfolds in nontargeted metabolite analyses. Network analyses revealed groups of interrelated amino acid, acylcarnitine, and fatty acid metabolites associated with all three newborn outcomes. In conclusion, cord blood metabolites are associated with newborn size and cord C-peptide, even after adjustment for maternal BMI and glucose during pregnancy. The paradoxical inverse association of metabolites with cord blood C-peptide suggests that the metabolite-insulin relationship reverses at a critical point in childhood to the well-described positive association seen in adolescents and adults. The well-known associations of branched chain amino acids and medium- and long-chain acylcarnitines with obesity in adolescents and adults appears to begin at birth and might serve as an early-life marker of obesity risk.