Serum nuclear magnetic resonance metabolomics analysis of human metastatic colorectal cancer: Biomarkers and pathway analysis.

We describe the use of nuclear magnetic resonance metabolomics to analyze blood serum samples from healthy individuals (n = 26) and those with metastatic colorectal cancer (CRC; n = 57). The assessment, employing both linear and nonlinear multivariate data analysis techniques, revealed specific metabolite changes associated with metastatic CRC, including increased levels of lactate, glutamate, and pyruvate, and decreased levels of certain amino acids and total fatty acids. Biomarker ratios such as glutamate-to-glutamine and pyruvate-to-alanine were also found to be related to CRC. The study also found that glutamate was linked to progression-free survival and that both glutamate and 3-hydroxybutyrate were risk factors for metastatic CRC. Additionally, gas chromatography coupled to flame-ionization detection was utilized to analyze the fatty acid profile and pathway analysis was performed on the profiled metabolites to understand the metabolic processes involved in CRC. A correlation was also found between the presence of certain metabolites in the blood of CRC patients and certain clinical features.

The Isolation of Specialty Compounds from Amphidinium carterae Biomass by Two-Step Solid-Phase and Liquid-Liquid Extraction.

The two main methods for partitioning crude methanolic extract from Amphidinium carterae biomass were compared. The objective was to obtain three enriched fractions containing amphidinols (APDs), carotenoids, and fatty acids. Since the most valuable bioproducts are APDs, their recovery was the principal goal. The first method consisted of a solid-phase extraction (SPE) in reverse phase that, for the first time, was optimized to fractionate organic methanolic extracts from Amphidinium carterae biomass using reverse-phase C18 as the adsorbent. The second method consisted of a two-step liquid-liquid extraction coupled with SPE and, alternatively, with solvent partitioning. The SPE method allowed the recovery of the biologically-active fraction (containing the APDs) by eluting with methanol (MeOH): water (H2O) (80:20 v/v). Alternatively, an APD purification strategy using solvent partitioning proved to be a better approach for providing APDs in a clear-cut way. When using n-butanol, APDs were obtained at a 70% concentration (w/w), whereas for the SPE method, the most concentrated fraction was only 18% (w/w). For the other fractions (carotenoids and fatty acids), a two-step liquid-liquid extraction (LLE) method coupled with the solvent partitioning method presented the best results.

NMR-based Metabolomics and Fatty Acid Profiles to Unravel Biomarkers in Preclinical Animal Models of Compulsive Behavior.

Compulsivity is a key manifestation of inhibitory control deficit and a cardinal symptom of psychopathological conditions such as obsessive-compulsive and attention-deficit hyperactivity disorders, in which metabolic alterations have raised attention as putative biomarkers for early identification. The present study assessed the metabolic profile in a preclinical model of a compulsive phenotype of rats. We used the schedule-induced polydipsia (SIP) method to classify male Wistar rats into high drinkers (HDs) or low drinkers (LDs) according to their compulsive drinking rate developed by exposure to a fixed-time 60 s (FT-60) schedule of reinforcement with water available ad libitum during 20 sessions. Before and after SIP, blood samples were collected for subsequent serum analysis by nuclear magnetic resonance spectroscopy coupled to multivariate analysis. Although no differences existed in the pre-SIP set, the compulsive drinking behavior induced remarkable metabolic alterations: HD rats selected by SIP exhibited a hyperlipidemic, hypoglycemic, and hyperglutaminergic profile compared with their low-compulsive counterparts. Interestingly, these alterations were not attributable to the mere exposure to reward pellets because a control experiment did not show differences between HDs and LDs after 20 sessions of pellet consumption without intermittent reinforcement. Our results shed light toward the implication of dietary and metabolic factors underpinning the vulnerability to compulsive behaviors.

Evaluation of ORAC, IR and NMR metabolomics for predicting ripening stage and variety in melon (Cucumis melo L.).

A combination of FTIR- and NMR-based metabolomics approaches coupled to multivariate data analysis techniques was used for the first time to characterize and explore the metabolic changes along the ripening of two Spanish melon landraces (Jimbee N and Jimbee XL). Furthermore, the fatty acids profile and the antioxidant capacity were investigated by GC-FID and ORAC method, respectively. Despite FTIR was able to identify changes in the sugars content between fruits of different maturity stages, it failed to discriminate between both varieties, being exceeded by NMR in both situations. Increased fruit maturation led to an increase of sucrose, fatty acids and ß-carotene, accompanied by a decrease of glucose, fructose, citrate, amino acids, and polyphenols (which were positively correlated with fruits antioxidant activity).

Effect of a Shading Mesh on the Metabolic, Nutritional, and Defense Profiles of Harvested Greenhouse-Grown Organic Tomato Fruits and Leaves Revealed by NMR Metabolomics.

Controlling the temperature inside a greenhouse during the summer is a problem of increasing importance in the Mediterranean countries, especially in the Spanish southeast. The metabolic profile of greenhouse tomatoes and leaves grown under conventional conditions and within the presence of a shade mesh (∼50% reduction of sunlight radiation) has been monitored. Tomatoes were weekly harvested from May to July 2017 and analyzed by NMR spectroscopy coupled to multivariate data analysis techniques, together with oxygen radical absorbance capacity (ORAC) assays (for antioxidant activity). Fatty acids and carotenoids profiles were unraveled by GC-FID and HPLC-DAD, respectively. To verify whether it would be possible to take advantage of different light growing conditions to potentiate a plant's defense system, leaves of the corresponding plants were collected and their methanolic extracts were analyzed by NMR toward deciphering new biomarkers, which were used to assess their antibacterial and antibiofilm activities. The presence of a shading mesh resulted in a reduction in tomato production and in smaller fruits with lower contents of sugars (glucose and fructose) and carotenoids (lycopene and ß-carotene) and higher contents of organic acids, amino acids, and polyunsaturated fatty acids (linoleic and oleic acids) and of phenylpropanoids and flavonoids (which contributed to an increased antioxidant activity). Methanolic extracts of leaves of nonshaded plants showed a higher antibiofilm activity than that from shaded plants. This activity was well-correlated with an increase of phenolic compounds, together with some specific amino acids and organic acids from tomato leaves.