Metabolic changes in antennal glands of Caribbean spiny lobsters Panulirus argus infected by Panulirus argus virus 1 (PaV1).

Panulirus argus virus 1 (PaV1) (Family Mininucleoviridae) causes chronic and systemic infection in wild juvenile spiny lobsters Panulirus argus (Latreille, 1804), ending in death by starvation and metabolic wasting. In marine decapods, the antennal gland is involved in osmoregulation and excretion. In this compact organ, fluid is filtered from the hemolymph, and ions are reabsorbed to produce a hypotonic urine. Although PaV1 is released with the urine in infected individuals, little is known regarding the metabolic effect of PaV1 in the antennal gland. The objective of this study was to perform a comparative evaluation of the metabolic profile of the antennal gland of clinically PaV1-infected lobsters versus those with no clinical signs of infection, using proton nuclear magnetic resonance analysis. Overall, 48 compounds were identified, and the most represented metabolites were those involved in carbohydrate, amino acid, energy, and nucleotide metabolism. Most of the metabolites that were down-regulated in the infected group were essential and non-essential amino acids. Some metabolites involved in the urea cycle and carbohydrate metabolism were also altered. This study represents a first approach to the metabolic evaluation of the antennal gland. We broadly discuss alterations in the content of several proteinogenic and non-proteinogenic amino acids and other key metabolites involved in energetic and nucleotide metabolism.

NMR Metabolic Profiling of Sargassum Species Under Different Stabilization/Extraction Processes.

The genus Sargassum is well represented by benthic and pelagic species, some of which form massive aggregates that can travel long distances due to the force of the ocean currents. Although they constitute an essential habitat for fish and invertebrate species, large accumulations of Sargassum in coastal areas generate several economic, environmental, and health impacts. It is important to recognize the species forming these aggregates, and identify the metabolites they produce, allowing for its exploitation, and therefore, better management practices. NMR metabolic profiling is a technique that can discriminate samples while detecting their unique or differential chemical features, and has been successfully used in the study and classification of several algal species. The present investigation studied the metabolic profiling of Sargassum species found on strandings at Puerto Morelos (Quintana Roo) east coast of the Mexican Caribbean. PCA of the 1 H-NMR profiles corresponding to S. natans, S. natans (morphotype VIII), S. fluitans, and a benthic Sargassum buxifolium allowed the discrimination of samples amongst them. Furthermore, discrimination between the two forms of S. natans was also possible. The PCA loading plot revealed that glutamine and glutamate have the highest influence in the clustering of the benthic Sargassum, while a high abundance of lactate, Myo-inositol, and trimethylamine is a unique feature from the S. natans morphotype VIII. Additional PLS-DA models showed that a heat-drying process improved the extraction of metabolites. Maceration and microwave-assisted extraction with water-ethanol led to similar profiles and thus any of them could be used in future investigations.

The use of 1 H-NMR Metabolomics to Optimise the Extraction and Preliminary Identification of Anthelmintic Products from the Leaves of Lysiloma latisiliquum.

INTRODUCTION: Tannin-rich forages are recognised as an important alternative for the control of gastrointestinal nematodes in small ruminants. Lysiloma latisiliquum, a forage commonly consumed by goats and sheep, has shown anthelmintic activity when tested against Haemonchus contortus. However, to date, the metabolites responsible for the activity are not known. OBJECTIVE: To use 1 H-NMR metabolomics in the extraction and identification of anthelmintic metabolites from L. latisiliquum. METHODOLOGY: Eight different solvent systems were compared for the optimum extraction of anthelmintic metabolites from L. latisiliquum. 1 H-NMR spectra of the tannin-free extracts were measured in methanol-d4 using trimethylsilylpropanoic acid (TSP) as internal standard. Extracts were also evaluated for their anthelmintic activity using the larval exsheathment inhibition assay against H. contortus. These data were correlated by multivariate analysis [principal component analysis (PCA) and orthogonal projections to latent structures discriminant analysis (OPLS-DA)] and analysed. To validate the results obtained after the OPLS-DA, a bioassay-guided isolation of bioactive metabolites was conducted. RESULTS: The PCA of the 1 H-NMR data allowed the identification of hydrophilic solvents as those best suited for the extraction of anthelmintics from L. latisiliquum and indicated that the bioactive metabolites are high-polarity, glycosylated products. Similarly, OPLS-DA of the data enabled the detection of activity-related signals, assigned to the glycosylated metabolites quercitrin and arbutin obtained from the bioassay-guided purification of the extract. CONCLUSION: The results of this investigation confirm metabolomics as a useful tool in the detection of bioactive metabolites in plants without previous phytochemical studies. Copyright © 2017 John Wiley & Sons, Ltd.

Characterization of cement mortars with regional organic and inorganic additives.

The present investigation establishes the basis for future studies in the southeast of México for the improvement of building materials by combining regional organic and inorganic nanoparticles in admixtures to formulate cement mortars with durability potential in structures of concrete. The characterization of the organic extract of Albzia tomentosa by nuclear magnetic resonance (NMR) revealed the presence of epicatechin (tannin related) and sucrose. Calcium zinc hydroxide dihydrate nanoparticles (CZ NPs) showed the highest surface area of 60.7 m2 g-1. The electrical resistivity, propagation of ultrasound velocity and water absorption by capillarity properties were individually evaluated for the organic extract, the inorganic nanoparticles and their admixtures in cement mortars, at a curing time of 7, 28 and 96 days with and optimal concentration of 5 mg mL-1 of the added additives. The best results were obtained at 96 days showing slightly but clear improvement of the electrical resistivity (23.40 ± 0.022 kΩ cm, 22.40 ± 0.004 kΩ cm and 22.29 ± 0.013 kΩ cm), propagation of ultrasound velocity (1370 ± 10 m s-1, 1345 m s-1 ± 6, 1310 ± 9 m s-1) and capillary coefficient (0.0044 kg m-2s-1/2, 0.0045 kg m-2s-1/2 and 0.0049 kg m-2s-1/2) properties of the cement mortars with CZ NPs, extraction Albizia solution (EAS) and CZ NPs + EAS respectively when compared to the mortar control (19.91 ± 0.036 kΩ cm, 1266 ± 15 m s-1 and 0.0082 kg m-2s-1/2).

1H-NMR Metabolomic Study of the Mushroom Pleurotus djamor for the Identification of Nematocidal Compounds.

Due to the increasing populations of anthelmintic-resistant gastrointestinal nematodes and as a consequence of the adverse effects of synthetic drugs, this study focuses on the search for secondary metabolites with nematocidal activity from the edible mushroom Pleurotus djamor using The proton nuclear magnetic resonance (1H-NMR) metabolomics. The highest activity was shown by the ethyl acetate fractions of mycelium (EC50 290.8 µg/mL) and basidiomes (EC50 282.7 µg/mL). Principal component analysis (PCA) and hierarchical data analysis (HCA) of the 1H-NMR metabolic profiles data showed that the ethanolic extracts, the ethyl acetate, butanol, and water fractions from mycelium have different metabolic profiles than those from basidiomes, while low polarity (hexane) fractions from both stages of fungal development show similar profiles. Orthogonal partial least squares discriminant analysis (OPLS-DA) allowed the identification of signals in the 1H-NMR metabolic profile associated with nematocidal activity. The signals yielded via OPLS-DA and bidimensional NMR analysis allowed the identification of uracil as a component in the ethyl acetate fraction from basidiomes, with an EC50 of 237.7 µg/mL. The results obtained showed that chemometric analyses of the 1H-NMR metabolic profiles represent a viable strategy for the identification of bioactive compounds from samples with complex chemical profiles.

Intra- and Interspecies Differences of Two Cecropia Species from Tabasco, Mexico, Determined through the Metabolic Analysis and 1H-NMR-Based Fingerprinting of Hydroalcoholic Extracts.

The genus Cecropia is used in the traditional medicine of Tabasco, Mexico, in diabetes and hypertension treatments, mainly without distinction of the species. This contribution aimed to carry out the metabolic analysis and Proton Nuclear Magnetic Resonance (1H-NMR) spectroscopy-based fingerprinting of the hydroalcoholic leaf extracts of Cecropia peltata (Cp) and Cecropia obtusifolia (Co) collected in five sub-regions of the State of Tabasco (Cp1, "Centro"; Cp2, "Chontalpa"; Cp3, "Pantanos"; Cp4, "Ríos" and Co5, "Sierra"). Firstly, the extracts were evaluated for their Total Phenol Content (TPC) and Total Flavonoid Content (TFC) by spectrophotometric methods. In addition, metabolic analysis was performed using High-Performance Liquid Chromatography with Diode-Array Detection HPLC-DAD, which allowed the quantification of the chemical markers: chlorogenic acid, isoorientin, and orientin, as well as a vitexin analog. Finally, metabolomic analysis was carried out based on the 1H-NMR spectra. The Cp4 extract (C. peltata from the "Ríos" sub-region) presented the highest values of TPC (155 ± 9.1 mg GAE/g E) and TFC (724 ± 22.2 mg RE/g E). The metabolic analysis was similar among the five samples; the highest concentrations of the four chemical markers were found in Cp3 (C. peltata from the "Pantanos" sub-region) for chlorogenic acid (39.8 ± 2.3 mg/g) and isoorientin (51.5 ± 2.9 mg/g), in Cp4 for orientin (49.9 ± 0.6 mg/g), and in Cp2 (C. peltata from the "Chontalpa" sub-region) for the vitexin analog (6.2 ± 0.2 mg/g). The metabolic analysis and the 1H-NMR fingerprint analysis showed intraspecies differences among the C. peltata samples and interspecies between C. peltata and C. obtusifolia, which were attributed to variations in the metabolite groups as well as in the proportion of sugars such as glucose and xylose.

1H NMR metabolomic analysis of skin and blubber of bottlenose dolphins reveals a functional metabolic dichotomy.

The common bottlenose dolphin (Tursiops truncatus) is a carnivorous cetacean that thrives in marine environments, one of the apex predators of the marine food web. They are found in coastal and estuarine ecosystems, which are known to be sensitive to environmental impacts. Dolphins are considered sentinel organisms for monitoring the health of coastal marine ecosystems due to their role as predators that can bioaccumulate contaminants. Although recent studies have focused on capturing the circulating metabolomes of these mammals, and in the context of pollutants and exposures in the marine environment, skin and blubber are important surface and protective tissues that have not been adequately probed for metabolism. Using a proton nuclear magnetic resonance spectroscopy (1H NMR) based metabolomics approach, we quantified 51 metabolites belonging to 74 different metabolic pathways in the skin and blubber of stranded bottlenose dolphin (n = 4) samples collected at different localities in the Southern Zone coast of Yucatan Peninsula of Mexico. Results indicate that metabolism of skin and blubber are quantitatively very different. These metabolite abundances could help discriminate the tissue-types using supervised partial least square regression discriminant analysis (PLSDA). Further, using hierarchical clustering analysis and random forest analysis of the metabolite abundances, the results pointed to unique metabolites that are important classifiers of the tissue-type. On one hand, the differential metabolic patterns, mainly linking fatty acid metabolism and ketogenic amino acids, seem to constitute a characteristic of blubber, thus pointing to fat synthesis and deposition. On the other hand, the skin showed several metabolites involved in gluconeogenic pathways, pointing towards an active anabolic energy-generating metabolism. The most notable pathways found in both tissues included: urea cycle, nucleotide metabolism, amino acid metabolism, glutathione metabolism among others. Our 1H NMR metabolomics analysis allowed the quantification of metabolites associated with these two organs, i.e., pyruvic acid, arginine, ornithine, 2-hydroxybutyric acid, 3-hydroxyisobutyric acid, and acetic acid, as discriminatory and classifying metabolites. These results would lead to further understanding of the functional and physiological roles of dolphin skin and blubber metabolism for better efforts in their conservation, as well as useful target biopsy tissues for monitoring of dolphin health conditions in marine pollution and ecotoxicology studies.

Effects of polyphenol removal methods on the in vitro exsheathment inhibitory activity of Lysiloma latisiliquum extracts against Haemonchus contortus larvae.

It has been suggested that the anthelmintic (AH) activity of tannin-rich forages is related to their tannin content. However, reports on tropical fodders such as Lysiloma latisiliquum describe the same AH activity after the addition of tannin-blocking agents, suggesting that the activity either depends on the method of tannin blocking/removal or is due to other type of secondary metabolites. This study compared both the effect of the drying process of the plant material and the effect of different polyphenol removal methods on the AH activity of L. latisiliquum acetone-water extracts when tested against Haemonchus contortus. The results showed that the extraction of oven-dried (OD) leaves of L. latisiliquum yielded a CT-free extract with high AH activity. However, polyphenol-free fractions showed similar or lower AH activity levels as of those original OD extract. HPLC analysis confirmed that common polyphenolic metabolites are not responsible for the AH activity of L. latisiliquum.

Antigiardial activity of flavonoids from leaves of Aphelandra scabra

AbstractAphelandra scabra (Vahl) Sm., Acanthaceae, is a shrub widely used by some Mayan communities as carminative, antidote, and remedy for some infections. Bio-guided isolation of the methanol extract of leaves led us to the purification of the anti-giardial metabolites cirsimaritin and sorbifolin, along with the inactive metabolites cirsimarin, sorbifolin-6-O-β-glucopyranoside, and squalene. Cirsimaritin displayed high activity in the anti-giardial bioassay with an IC50 = 3.8 μM, being considered as outstanding when compared to previous reported metabolites, while sorbifolin showed a low activity with an IC50 = 75.6 μM. Additionally, both compounds proved not to be cytotoxic in an in vitro bioassay against HEK-293, a normal cell line. This is the first investigation on anti-giardial properties of A. scabra and its phytochemistry as well, thus the isolated compounds are considered as new for the plant genus and for the species.