Revealing the 1H-NMR Profiling of Six Edible Mushrooms Consumed in the Northeastern Highlands of Puebla, Mexico.

The 1H-NMR metabolomics profiling of six edible mushrooms consumed in the northeastern highlands of Puebla, Mexico is presented. These fungi were morpho- and molecularly identified as Infundibulicybe squamulosa, Amanita jacksonii, Lepista nuda, Russula delica, Russula brevipes, and Lactarius indigo. The chemical profiling confirmed the presence of eight essential amino acids and their derivatives, six organic acids, six nucleosides, low amounts of reducing sugars, and valuable nutraceuticals such as betaine, carnitine, glycero-3-phosphocholine and O-acetylcarnitine which were differentially determined and quantified in the six mushrooms by qNMR. Principal component analysis (PCA) and orthogonal projections to latent structures discriminant analysis (OPLS-DA) generated four different groups. Two of these groups were constituted by fungal species with phylogenic relationships whereas non-phylogenetic related species were separated from each other. The potential use of 1H-NMR metabolomics and chemometrics to group macromycetes and determine the nutritional and nutraceutical potential of these local foods is demonstrated.

Exploring the chemical diversity of Capsicum chinense cultivars using NMR-based metabolomics and machine learning methods.

The habanero pepper (Capsicum chinense) is a prominent spicy fruit integral to the historical, social, cultural, and economic fabric of the Yucatan peninsula in Mexico. This study leverages the power of 1H NMR spectroscopy coupled with machine learning algorithms to dissect the metabolomic profile of eleven C. chinense cultivars, including those grown by INIFAP (Habanero-Jaguar, Antillano-HRA 1-1, Antillano-HRA 7-1, Habanero-HAm-18A, Habanero-HC-23C, and Jolokia-NJolokia-22) and commercial hybrids (Habanero-Rey Votán, Habanero-Kabal, Balam, USAPR10117, and Rey Pakal). A total of fifty metabolites, encompassing sugars, amino acids, short-chain organic acids, and nucleosides, were identified from the 1H NMR spectra. The optimized machine learning model proficiently predicted the similarity percentage between the INIFAP-grown cultivars and commercial hybrids, thereby facilitating a comprehensive comparison. Biomarkers unique to each cultivar were delineated, revealing that the Habanero-Rey Votán cultivar is characterized by the highest concentration of sugars. In contrast, the Balam cultivar is rich in amino acids and short-chain organic acids, sharing a similar metabolomic profile with the Jolokia-NJolokia-22 cultivar. The findings of this study underscore the efficacy and reliability of NMR-based metabolomics as a robust tool for differentiating C. chinense cultivars based on their intricate chemical profiles. This approach not only contributes to the scientific understanding of the metabolomic diversity among habanero peppers but also holds potential implications for food science, agriculture, and the culinary arts.

NMR-based metabolomics to determine the fluctuation of metabolites in hydroponic purslane crops at different harvesting times.

Purslane (Portulaca oleracea L.) has a high content of nutrients and medicinal effects that depend on the genotype, harvesting time, and production system. The objective of the present research work was to elucidate the NMR-based metabolomics profiling of three native purslane cultivars from Mexico (Xochimilco, Mixquic, and Cuautla) grown under hydroponic conditions and harvested in three different times (32, 39, and 46 days after emergence). Thirty-nine metabolites identified in the 1H NMR spectra of aerial parts of purslane, 5 sugars, 15 amino acids, 8 organic acids, 3 caffeoylquinic acids, as well as 2 alcohols and 3 nucleosides, choline, O-phosphocholine and trigonelline were also detected. A total of 37 compounds were detected in native purslane from Xochimilco and Cuautla, whereas 39 compounds were detected in purslane from Mixquic. Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) separated the cultivars into three clusters. Mixquic cultivar had the highest number of differential compounds (amino acids and carbohydrates), followed by Xochimilco and Cuautla cultivars, respectively. Changes in the metabolome were observed in latest times of harvest for all the cultivars studied. The differential compounds were glucose, fructose, galactose, pyruvate, choline, and 2-hydroxysobutyrate. The results obtained in this investigation may contribute to selecting the best cultivar of purslane and the best time in which the levels of nutrients are optimal.

1H NMR Profiling of the Venom from Hylesia continua: Implications of Small Molecules for Lepidopterism.

Lepidopterism caused by caterpillar contact is considered a public health problem around the world. The local and systemic responses of this pathology include short- and long-term inflammatory events. Although the proteolytic activity of the venoms from caterpillars is strongly associated with an inflammatory response in humans and murine models, fast and acute symptoms such as a burning sensation, itching, and pain should be related to the presence of low-weight hydrophilic molecules which easily influence cell metabolism. This investigation reports on the 1H-Nuclear Magnetic Resonance (NMR) profiling of the venom from the larva of Hylesia continua, a caterpillar linked to frequent cases of lepidopterism in the northern highlands of Puebla, Mexico. According to one-dimensional (1D) and two-dimensional (2D) NMR data, the venom of H. continua contained 19 compounds with proven pain-inducing activity (i.e., acetic acid, lactic acid, formic acid, succinic acid, 2-hydroxyglutaric acid, ethanol, and glutamate), inflammatory activity (i.e., cadaverine, putrescine, and acetoin), as well as natural immunosuppressive activity (i.e., O-phosphocholine and urocanic acid). The levels of the 19 compounds were calculated using quantitative-NMR (qNMR) and extensively discussed on the basis of their toxic properties which partially explain typical symptoms of lepidopterism caused by the larvae of H. continua. To the best of our knowledge, this is the first investigation reporting a complex mixture of small molecules with inflammatory properties dissolved in the venom of a lepidopteran larva.

Metabolomic Insights of Biosurfactant Activity from Bacillus niabensis against Planktonic Cells and Biofilm of Pseudomonas stutzeri Involved in Marine Biofouling.

In marine environments, biofilm can cause negative impacts, including the biofouling process. In the search for new non-toxic formulations that inhibit biofilm, biosurfactants (BS) produced by the genus Bacillus have demonstrated considerable potential. To elucidate the changes that BS from B. niabensis promote in growth inhibition and biofilm formation, this research performed a nuclear magnetic resonance (NMR) metabolomic profile analysis to compare the metabolic differences between planktonic cells and biofilms of Pseudomonas stutzeri, a pioneer fouling bacteria. The multivariate analysis showed a clear separation between groups with a higher concentration of metabolites in the biofilm than in planktonic cells of P. stutzeri. When planktonic and biofilm stages were treated with BS, some differences were found among them. In planktonic cells, the addition of BS had a minor effect on growth inhibition, but at a metabolic level, NADP+, trehalose, acetone, glucose, and betaine were up-regulated in response to osmotic stress. When the biofilm was treated with the BS, a clear inhibition was observed and metabolites such as glucose, acetic acid, histidine, lactic acid, phenylalanine, uracil, and NADP+ were also up-regulated, while trehalose and histamine were down-regulated in response to the antibacterial effect of the BS.

1 H-NMR Metabolomics Profiling and Volatile Content of 'Hoja Santa' (Piper auritum Kunth): A Millenary Edible Plant Consumed in Mexico.

The leaves of Piper auritum Kunth ('Hoja Santa') have been consumed for centuries by native people of central and southern Mexico as a fresh vegetable or condiment. Herein we present the result of the 1 H-NMR metabolomics profiling of three accessions of P. auritum harvested in three different provinces of Mexico (Puebla, Tlaxcala, and Oaxaca). The volatile content associated with the flavoring properties of the plant was also determined by GC/MS. The non-targeted metabolome of these samples revealed that P. auritum is a source of free essential amino acids such as isoleucine, leucine, threonine, valine, histidine, phenylalanine, and tryptophan as well as organic acids, free monosaccharides, and valuable nutraceuticals such as trigonelline, Myo-inositol, betaine, and choline. Principal component analysis and orthogonal partial least squares discriminated analysis of the metabolites found in P. auritum revealed trigonelline as the main differential compound found in the three studied accessions, suggesting this metabolite as a possible chemical marker. According to these statistical approaches, 60 % of the differential metabolites were provided by Oaxaca samples, suggesting that leaves harvested in this province have better (p<0.05) nutritional properties than the other samples analyzed. Nevertheless, the high abundance of the anti-nutrient safrole (90 %) in the volatile fraction, advises the potential toxicity of P. auritum consumed in Oaxaca. On the other hand, samples harvested in the northern highlands of Puebla, contained the lowest levels of safrole (30 %) and acceptable levels of nutrients and nutraceuticals including choline. From the three groups of studied plants, those harvested in the northern highlands from Puebla, could be considered safer for human consumption than the other analyzed accessions.

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.

Fungistatic Films Containing Cinnamon Essential Oil: New Coatings to Preserve the Nutraceutical Content of Avocado Fruit against Fusariosis.

Avocadoes are a rich source of nutrients and nutraceuticals that preserve human health. Nevertheless, this fruit is susceptible to phytopathogen infection during the postharvest period causing severe economic losses. Herein, we report on the in situ antifungal assessment of biodegradable films impregnated with the essential oil of Cinnamomum verum (CvEO) as natural fungistatic coatings to extend postharvest quality of Hass avocadoes (Persea americana cv. Hass). These coatings were evaluated on fruits previously infected with a native strain of Fusarium verticillioides. The cytotoxic assessment of CvEO on F. verticillioides revealed a minimum inhibitory concentration of 0.3±0.0 g L-1 whereas its chemical profiling showed (E)-cinnamaldehyde (45.9 %) 1,3,8-p-menthatriene (7.9 %) and linalool (6.8 %) as the major bioactive compounds. Four coatings (FC1-FC4) were made using chitosan (1 %) and a diverse quantity of CvEO (0.4-1.3 % w/v). The physicochemical properties demonstrated that the films FC3 and FC4 have the optimal characteristics of a food coating. Avocadoes preserved with the films FC3 and FC4 substantially kept (p<0.01) fruit firmness and the nutraceutical content of infected avocados at least for 21 d. The results of the present investigation suggest that our hybrid materials can conserve basic nutritional parameters such as fiber, protein, reducing sugars, as well as the content of unsaturated fatty acids which are the main nutraceuticals of this fruit.

Bean cultivars (Phaseolus vulgaris L.) under the spotlight of NMR metabolomics.

The seeds of Phaseolus vulgaris are a rich source of protein consumed around the world and are considered as the most important source of proteins and antioxidants in the Mexican diet. This work reports on the 1H NMR metabolomics profiling of the cultivars Peruano (FPe), Pinto (FPi), Flor de mayo (FM), Negro (FN) and Flor de junio (FJ). Total phenolics, total flavonoids and total protein contents were determined to complement the nutritional facts in seeds and leaves. According to our results, the metabolomics fingerprint of beans seeds and leaves were very similar, showing the presence of 52 metabolites, 46 in seeds and 48 in leaves, including 8 sugars, 17 amino acids, 15 organic acids, 5 nucleosides and 7 miscellaneous compounds. In seeds, free amino acids were detected in higher concentrations than in the leaves, whereas organic acids were more abundant in leaves than in seeds. With multivariate and cluster analysis it was possible to rank the cultivars according to their nutritional properties according to NMR profiling, then a machine learning algorithm was used to reveal the most important differential metabolites which are the key for correct classification. The results coincide in highlighting the FN seeds and FPe leaves for the best nutritional facts. Finally, in terms of cultivars, FN and FM present the best nutritional properties, with high protein and flavonoids content, as well as, a high concentration of amino acids and nucleosides.

Aberrant sphingomyelin 31P-NMR signatures in giant cell tumour of bone.

An understanding of the biochemistry of the giant cell tumour of bone (GCTB) provides an opportunity for the development of prognostic markers and identification of therapeutic targets. Based on metabolomic analysis, we proposed glycerophospholipid metabolism as the altered pathway in GCTB., The objective of this study was to identify these altered metabolites. Using phosphorus-31 nuclear magnetic resonance spectroscopy (31P-NMR), sphingomyelin was determined to be the most dysregulated phospholipid in tissue samples from six patients with GCTB. Enzymes related to its biosynthesis and hydrolysis were examined using immunodetection techniques. High expression of sphingomyelin synthases 1 and 2, but low expression of neutral sphingomyelinase 2 (nSMase2) was found in GCTB tissues compared to non-neoplastic bone tissues. Sphingomyelin/ceramide biosynthesis is dysregulated in GCTB due to alterations in the expression of SMS1, SMS2, and nSMase2.

Antitumor Effect of Zwitterions of Imidazolium Derived from L-methionine in BALB/c Mice with Lymphoma L5178Y.

BACKGROUND: In the therapy of cancer, several treatments have been designed using nanomaterials, among which gold nanoparticles (AuNPs) have been featured as a promising antitumoral agent. Our research group has developed the synthesis of gold nanoparticles L-AuNPs and D-AuNPs stabilized with zwitterions of imidazolium (L-1 and D-1) derived from L-methionine and D-methionine. Because the stabilizer agent is chiral, we observed through circular dichroism that AuNPs also present chirality; such chirality as well as the fact that the stabilizing agent contains fragments of methionine and imidazolium that are commonly involved in biological processes, opens up the possibility that this system may have biological compatibility. Additionally, the presence of methionine in the stabilizing agent opens the application of this system as a possible antitumor agent because methionine is involved in methylation processes of molecules such as DNA. OBJECTIVE: The aim of this research is the evaluation of the antitumor activity of gold nanoparticles stabilized with zwitterions of imidazolium (L-AuNPs) derived from L-methionine in the model of BALB/c mice with lymphoma L5178Y. METHODS: Taking as a parameter cell density, the evaluation of the inhibitory effect of L-AuNPs was carried out with a series of in vivo tests in BALB/c type mice; three groups of five mice each were formed (Groups 1, 2 and 3); all mice were i.p. inoculated with the lymphoblast murine L5178Y. Group 1 consisted of mice without treatment. In the Groups 2 and 3 the mice were treated with L-AuNPs at 0.3 mg/Kg on days 1, 7 and 14 by orally and intraperitonally respectively. RESULTS: These results show low antitumor activity of these gold nanoparticles (L-NPsAu) but interestingly, the imidazolium stabilizing agent of gold nanoparticle (L-1) displayed promising antitumor activity. On the other hand, the enantiomer of L-1, (D-1) as well as asymmetric imidazole derivate from L-methionine (L-2), do not exhibit the same activity as L-1. CONCLUSION: The imidazolium stabilizing agent (L-1) displayed promising antitumor activity. Modifications in the structure of L-1 showed that, the stereochemistry (like D-1) and the presence of methionine fragments (like L-2) are determinants in the antitumor activity of this compound.

Experimental races of Capsicum annuum cv. jalapeño: Chemical characterization and classification by 1H NMR/machine learning.

This work reports on the metabolic fingerprinting of ten new races of Capsicum annuum cv. jalapeño using 1H NMR based metabolomics coupled to machine learning projections. Ten races were classified and evaluated according to their differential metabolites, variables of commercial interest and by multivariate data analysis/machine learning algorithm. According to our results, experimental races of jalapeño peppers exhibited differences in carbohydrate, amino acid, nucleotide and organic acid contents. Forty-eight metabolites were identified by 1D and 2D NMR and the differential metabolites were quantified by qNMR. Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) separated the studied races into two groups. The group A included the races Colosus, Emperador, Fundador and Rayo whereas the group B included the races Don Benito, SMJ 1416, SMJ 1417, SMJ 1423, SMJ 145 and STAM J0904. OPLS-DA revealed that levels of citric acid in group A were higher than in group B, while the levels of asparagine, fumaric acid, GABA, glucose, malic acid, pyruvic, quinic acid, sucrose and tryptophan were higher in the group B. Remarkably, ascorbic acid was exclusively found in the race Colosus. Random forest model revealed the diversity of the experimental races and the similarity rate with the well-established races. The most relevant variables used to generate a model were length, weight, yield, width, xylose content and organic acids content.

Differentiating, evaluating, and classifying three quinoa ecotypes by washing, cooking and germination treatments, using 1H NMR-based metabolomic approach.

We processed three quinoa ecotypes as they are commonly consumed in a daily diet. For the treatments, quinoa seeds were washed, cooked, and/or germinated. Following treated, we used 1H NMR-based metabolomic profiling to explore differences between the ecotypes. Then, for a non-targeted and targeted food fingerprint analysis of samples, we performed multivariable data analyses, including principal component analysis (PCA), orthogonal partial least squares discriminant analysis (OPLS-DA), and hierarchical cluster analysis. From our study, we were able to discriminate each quinoa ecotype regardless of treatment based on its metabolomic profiling. Additionally, we were able to identify 30 metabolites that were useful to determine the effect of each treatment on nutritional composition. Germination increased the content of most metabolites irrespective of ecotype. In general, ecotype CQE_03 was different from ecotypes CQE_01 and CQE_02. Our phytochemical analysis revealed the effects of washing, cooking, and/or germination, particularly on saponins content.

Revealing the 1H NMR metabolome of mirasol chili peppers (Capsicum annuum) infected by Candidatus Phytoplasma trifolii.

The infection of Capsicum annuum cv. mirasol by Candidatus Phytoplasma trifolii (16SrVI) causes devastating crop losses in northern Mexico. This study addresses the metabolomics profiling of mirasol chili peppers (Capsicum annuum cv. mirasol) infected by Candidatus Phytoplasma trifolii. For this study, 25 diseased fruits and 25 healthy fruits were used. Principal component analysis (PCA) and orthogonal projections to latent structures discriminant analysis (OPLS-DA) revealed dramatic changes in the content of 42 metabolites which were identified in diseased and healthy mirasol chili peppers. The endogenous levels of fructose, glucose and formic acid were substantially decreased in the diseased chili peppers. In the same group of samples, high concentrations of alanine, asparagine, fumaric acid, sucrose and threonine were observed. The content of Choline didnt present a significant difference. This evidence supports the fact that Candidatus Phytoplasma trifolii infection reduces de CO2 fixation into carbohydrates, decreases invertase activity, and inhibits glycolysis in the diseased plant tissues. The levels of ascorbic acid, capsaicin and dihydrocapsaicinin in diseased fruits were dramatically decreased, suggesting that Candidatus Phytoplasma trifolii can reduce the pungency and the nutraceutical value of mirasol chili peppers.

1 H-NMR metabolomics profiling of recombinant tobacco plants holding a promoter of a sesquiterpene cyclase.

INTRODUCTION: Nicotiana tabacum is a plant model intensively used in the bio-engineering pharmaceutical industry as a platform to produce drugs and therapeutic agents. Currently, no information regarding the non-targeted metabolome of transgenic tobacco containing recombinant regulatory sequences is available. OBJECTIVE: To compare the proton nuclear magnetic resonance (1 H-NMR) metabolomics profiling of a recombinant Nicotiana tabacum strain containing a promoter of a sesquiterpene cyclase from Capsicum annuum driving GUS expression, versus wild-type samples. Methodology The non-targeted 1 H-NMR metabolome was obtained and processed by principal component analysis (PCA) and orthogonal projection to latent structures discriminant analysis (OPLS-DA). The differential metabolites were quantified by quantitative NMR. RESULTS: PCA and OPLS-DA revealed 37 metabolites including 16 discriminant compounds for transgenic samples. Ethanol (0.4 mg g-1 ), the main differential compound, was exclusively detected in transgenic tobacco; however, high levels of formate (0.28 mg g-1 ) and acetate (0.3 mg g-1 ) were simultaneously observed in the same group of samples. Cembratriene-4,6-diol, an antitumour and neuroprotective compound, and capsidiol, a known phytoalexin, increased by about 30% in transgenic samples. In addition, the endogenous levels of the antioxidant caffeoylquinic acid isomers increased by 50% in comparison to those of wild-type tobaccos. CONCLUSION: Our results support the occurrence of metabolic differences between wild type and transgenic tobacco containing a promoter of a Capsicum sesquiterpene cyclase gene. Interestingly, the recombinant transgenic strain studied accumulated high amounts of added value compounds with biological activity.

Metabolic profiling of serum in patients with cartilage tumours using 1 H-NMR spectroscopy: A pilot study.

Cartilage-forming lesions include tumours that can vary in severity from benign enchondromas to high-grade malignant chondrosarcomas. Chondrosarcoma is the second most frequent malignant bone tumour, accounting for 20-30% of all malignant bone neoplasms. Surgery is the standard treatment for cartilage tumours (CTs); however, their incidental diagnosis and the difficult differentiation of low-grade lesions like chondrosarcoma grade I from benign entities like enchondroma are challenges for clinical management. In this sense, the search for circulating biomarkers for early detection and prognosis is an ongoing interest. Targeted metabolomics is a powerful tool that can propose potential biomarkers in biological fluids as well as help to discover disturbed metabolic pathways to reveal tumour pathogenesis. In this context, the aim of this study was to investigate the 1 H nuclear magnetic resonance metabolomic serum profile of patients with CTs contrasted with healthy controls. Forty-one metabolites were identified and quantified; the multivariate statistical methods principal component analysis and partial least squares discriminant analysis reveal a clear separation of the CT group, that is, the differential metabolites that were involved in two main metabolic pathways: the taurine and hypotaurine metabolism and synthesis and degradation of ketone bodies. Our results represent preliminary work for emergent serum-based diagnostics or prognostic methods for patients with chondrogenic tumours.

1H NMR-based fingerprinting of eleven Mexican Capsicum annuum cultivars.

Approximately 90% of the chili peppers consumed in the world are harvested in Mexico. The present article describes the untargeted 1H NMR-based metabolomic profiling of 11 cultivars of Capsicum annuum species which are routinely consumed worldwide. The metabolomic fingerprinting detected via 1H NMR contained 44 metabolites including sugars, amino acids, organic acids, polyphenolic acids and alcohols which were identified by comparison with the literature data, with Chenomx database and by 2D NMR. Statistical approaches based on principal component analysis (PCA) and linear discriminant analysis (LDA) were used to classify the Capsicum annuum cultivars according to their metabolite profile. LDA revealed metabolomic differences and similarities among Capsicum annuum cultivars, whereas hierarchical cluster analysis (HCA) significantly separated the cultivars according to the phylogenetic trees obtained. Substantial endogenous levels of free amino acids and carbohydrates were detected in all the studied cultivars but interestingly, Capsicum annuum cv. mirasol and C. annuum cv. chilaca contained almost three-fold more endogenous levels of vitamin C than the other cultivars. Considering that this antioxidant was found in crude aqueous extracts, its abundance could be directly proportional to its bioavailability for human nutrition. The results suggest that 1H NMR is an effective method to determine differences among cultivars of the Capsicum annuum species.

1H NMR-based metabolomics profiling of ten new races from Capsicum annuum cv. serrano produced in Mexico.

Herein we report on the 1H NMR-based metabolomics profiling of ten new races of Capsicum annuum cv. serrano, cultivated in Mexico. Forty eight metabolites (including sugars, amino acids, organic acids, polyphenolic acids and alcohols) were identified and quantified by 2D NMR and qNMR, respectively. Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) separated the ten races into two clusters, from which citric acid, formic acid, fumaric acid, malic acid, glucose, fructose, sucrose and galactose were found as differential metabolites. This is the first study describing the chemical profiling of ten new races of Capsicum annuum cv. serrano and the spectrometric method used presently is characterized by great simplicity, robustness and reproducibility. Thus, this technique can be used for establishing reliable metabolomic fingerprints of different races of Capsicum annuum cv. serrano.

1 H-NMR Based Metabolomics Profiling of Citrus Juices Produced in Veracruz, México.

This study describes the 1 H-NMR-based metabolomics profiling of juices from citrus fruits harvested in the state of Veracruz, México. The hydrophilic profile of commercial lemons (Agrio and Persian), tangerines (Fremont and Mónica), oranges (Valencia and Washington Navel), and grapefruits (Red Ruby and Rio Red) was determined. According to our results, 35 metabolites were identified in the 1 H-NMR profiling. The statistical differences obtained by PCA and OPLS-DA revealed that specific amino acids, sugars, and organic acids were differential metabolites in the species and cultivars studied. High endogenous levels of sucrose (10-190 mM), α-glucose, ß-glucose (α- and ß-isomers, 40-205 mm), and fructose (36-170 mm) were detected in the juices of grapefruits, oranges, and tangerines, whereas citric acid (40-530 mm) was the principal organic acid in the juices of lemons. To calculate the specific amounts of metabolites from these species and their cultivars, the results were finely analyzed using the qNMR method. According to these calculations, Valencia oranges had the highest concentration of ascorbic acid (>2 mm). The described 1 H-NMR method is highly reproducible, inexpensive, and highly robust in comparison to other analytical methods used to determine the hydrophilic profile of citrus juices.

Synthesis and In Vitro Evaluation of Tetrahydroquinoline Derivatives as Antiproliferative Compounds of Breast Cancer via Targeting the GPER.

BACKGROUND: Some reports have demonstrated the role of the G Protein-coupled Estrogen Receptor (GPER) in growth and proliferation of breast cancer cells. OBJECTIVE: In an effort to develop new therapeutic strategies against breast cancer, we employed an in silico study to explore the binding modes of tetrahydroquinoline 2 and 4 to be compared with the reported ligands G1 and G1PABA. METHODS: This study aimed to design and filter ligands by in silico studies determining their Lipinski's rule, toxicity and binding properties with GPER to achieve experimental assays as anti-proliferative compounds of breast cancer cell lines. RESULTS: In silico studies suggest as promissory two tetrahydroquinoline 2 and 4 which contain a carboxyl group instead of the acetyl group (as is needed for G1 synthesis), which add low (2) and high hindrance (4) chemical moieties to explore the polar, hydrophobic and hindrance effects. Docking and molecular dynamics simulations of the target compounds were performed with GPER to explore their binding mode and free energy values. In addition, the target small molecules were synthesized and assayed in vitro using breast cancer cells (MCF-7 and MDA-MB-231). Experimental assays showed that compound 2 decreased cell proliferation, showing IC50 values of 50µM and 25µM after 72h of treatment of MCF-7 and MDA-MB-231 cell lines, respectively. Importantly, compound 2 showed a similar inhibitory effect on proliferation as G1 compound in MDA-MB-231 cells, suggesting that both ligands reach the GPER-binding site in a similar way, as was demonstrated through in silico studies. CONCLUSION: A concentration-dependent inhibition of cell proliferation occurred with compound 2 in the two cell lines regardless of GPER.