Study of the Stability of Wine Samples for 1H-NMR Metabolomic Profile Analysis through Chemometrics Methods.

Wine is a temperature, light, and oxygen-sensitive product, so its physicochemical characteristics can be modified by variations in temperature and time when samples are either sampled, transported, and/or analyzed. These changes can alter its metabolomic fingerprinting, impacting further classification tasks and quality/quantitative analyses. For these reasons, the aim of this work is to compare and analyze the information obtained by different chemometric methods used in a complementary form (PCA, ASCA, and PARAFAC) to study 1H-NMR spectra variations of four red wine samples kept at different temperatures and time lapses. In conjunction, distinctive changes in the spectra are satisfactorily tracked with each chemometric method. The chemometric analyses reveal variations related to the wine sample, temperature, and time, as well as the interactions among these factors. Moreover, the magnitude and statistical significance of the effects are satisfactorily accounted for by ASCA, while the time-related effects variations are encountered by PARAFAC modeling. Acetaldehyde, formic acid, polyphenols, carbohydrates, lactic acid, ethyl lactate, methanol, choline, succinic acid, proline, acetoin, acetic acid, 1,3-propanediol, isopentanol, and some amino acids are identified as some of the metabolites which present the most important variations.

Two-photon isomerization properties of donor-acceptor Stenhouse adducts.

Donor-acceptor Stenhouse adducts (DASAs) are important photo-responsive molecules that undergo electrocyclic reactions after light absorption. From these properties, DASAs have received extensive attention as photo-switches with negative photochromism. Meanwhile, several photochemical applications require isomerization events to take place in highly localized volumes at variable depths. Such focused photoreactions can be achieved if the electronic excitation is induced through a non-linear optical process. In this contribution we describe DASAs substituted with extended donor groups which provide them with significant two-photon absorption properties. We characterized the photo-induced transformation of these DASAs from the open polymethinic form to their cyclopentenic isomer with the use of 800 nm femtosecond pulses. These studies verified that the biphotonic excitation produces equivalent photoreactions as linear absorbance. We also determined these DASAs' two-photon absorption cross sections from measurements of their photoconverted yield after biphotonic excitation. As we show, specific donor sections provide these systems with important biphotonic cross-sections as high as 615 GM units. Such properties make these DASAs among the most non-linearly active photo-switchable molecules. Calculations at the TDDFT level with the optimally tuned range-separated functional OT-CAM-B3LYP, together with quadratic response methods indicate that the non-linear photochemical properties in these molecules involve higher lying electronic states above the first excited singlet. This result is consistent with the observed relation between their two-photon chemistry and the onset of their short wavelength absorption features around 400 nm. This is the first report of the non-linear photochemistry of DASAs. The two-photon isomerization properties of DASAs extend their applications to 3D-photocontrol, non-linear lithography, variable depth birefringence, and localized drug delivery schemes.

Microanalytical Characterization of an Innovative Modern Mural Painting Technique by SEM-EDS, NMR and Micro-ATR-FTIR among Others.

During the 20th century, modern painters experimented with different mediums and painting techniques, one of them was Rafael Coronel in his mural painting, Paisaje Abstracto (Abstract landscape). The painting was created with a peculiar pouring technique and an unknown binding medium; ageing produced fractures and severe conservation problems. Therefore, the characterization of the painting medium became an urgent matter in order to understand the current condition of the painting and to develop a proper treatment. The aim of this research was to characterize the chemical composition and painting technique of Paisaje Abstracto. To approach this goal two microsamples were taken and analyzed by optical microscopy (OM), scanning electron microscopy (SEM) with energy dispersive spectroscopy (EDS), nuclear magnetic resonance (NMR) spectroscopy, attenuated total reflection Fourier transform infrared spectroscopy (ATR-FTIR), micro attenuated total reflection Fourier transform infrared spectroscopy (micro-ATR-FTIR) and gas chromatography/mass spectrometry (GC/MS). The analysis allowed for the identification of cadmium sulfide (CdS) and titanium dioxide (TiO2) as inorganic pigments; aluminosilicate fillers; poly(methyl methacrylate) (pMMA) as a binder; MMA monomer, red organic pigment PR181; benzoyl peroxide, dibutyl phthalate and 1-octadecanol as organic additives. This study presents an innovative painting technique with pMMA, a medium not commonly used by artists, which was probably polymerized onto the painting support.

A Longitudinal 1H NMR-Based Metabolic Profile Analysis of Urine from Hospitalized Premature Newborns Receiving Enteral and Parenteral Nutrition.

Preterm newborns are extremely vulnerable to morbidities, complications, and death. Preterm birth is a global public health problem due to its socioeconomic burden. Nurturing preterm newborns is a critical medical issue because they have limited nutrient stores and it is difficult to establish enteral feeding, which leads to inadequate growth frequently associated with poor neurodevelopmental outcomes. Parenteral nutrition (PN) provides nutrients to preterm newborns, but its biochemical effects are not completely known. To study the effect of PN treatment on preterm newborns, an untargeted metabolomic 1H nuclear magnetic resonance (NMR) assay was performed on 107 urine samples from 34 hospitalized patients. Multivariate data (Principal Component Analysis, PCA, Orthogonal partial least squares discriminant analysis OPLS-DA, parallel factor analysis PARAFAC-2) and univariate analyses were used to identify the association of specific spectral data with different nutritional types (NTs) and gestational ages. Our results revealed changes in the metabolic profile related to the NT, with the tricarboxylic acid cycle and galactose metabolic pathways being the most impacted pathways. Low citrate and succinate levels, despite higher glucose relative urinary concentrations, seem to constitute the metabolic profile found in the studied critically ill preterm newborns who received PN, indicating an energetic dysfunction that must be taken into account for better nutritional management.

NMR and other molecular and elemental spectroscopies for the characterization of samples from an outdoor mural painting by Siqueiros.

This study presents the use of high resolution Nuclear Magnetic Resonance (NMR) as an outstanding tool for identifying with accuracy the structure of the polymers used as binders and the additives used as plasticizers in samples from a mural painted by David Alfaro Siqueiros between 1964 and 1972, an important period in which his artistic production has not been previously studied. Additionally, the presence of some organic pigments was detected. The mural Trazos de composición piramidal currently located outside of Siqueiros' studio La Tallera, in Cuernavaca Morelos (Mexico) was an artistic experiment done by Siqueiros to test outdoor painting. Eight micro-samples of paint were thoroughly analyzed by Optical Microscopy (OM), Scanning Electron Microscopy - Energy Dispersive X-Ray Spectroscopy (SEM-EDS), Fiber Optics Reflectance Spectra (FORS), Transmission Fourier Transform Infrared Spectroscopy with Attenuated Total Reflectance mode (FTIR-ATR) and Pyrolysis-Gas Chromatography/Mass Spectrometry (Py-GC/MS). Through these studies, contrasted with an historical investigation, we demonstrate that the samples present a multi-layered and complex painting system because Siqueiros made at least two versions of the painting. The multi-layered structure and the variable composition of each layer challenged the application of several analytical methodologies traditionally used for cultural heritage materials. In one sample, each layer was separated and they were studied in detail by FTIR-ATR, Py-GC/MS and NMR. This study revealed the presence of two main binders: acrylics in the first layers and nitrocellulose in the last one, also some phthalates and fatty acids were detected as additives. Inorganic pigments and fillers were identified as titanium dioxide and iron red oxide as well as diatoms and carbonates. The main contribution of this study, where NMR spectroscopy was applied for the first time in modern mural painting analysis, is the characterization of the materials used by Siqueiros during the last years of his artistic production.

1H -NMR Metabolomics Study of the Effect of Cisplatin and Casiopeina IIgly on MDA-MB-231 Breast Tumor Cells.

The knowledge of the metabolic processes of designed metallodrugs for cancer treatment is an area that has been not profoundly studied. Casiopeina IIgly (CasIIgly), which belongs to the Casiopeínas® family, is a copper (II) coordination compound that has shown good biological activity against several cancer cells, low toxicity in normal cells, and antineoplastic activity in in vivo murine and xenografted models. In this work we employed a triple-negative highly metastatic breast carcinoma line (MDA-MB-231), which is one of the cancer types with a great mortality index, for 1H-NMR metabolomic analysis using cisplatin and CasIIgly, in order to quantify the effect of metallodrugs in the metabolic profile of this cell tumor line as a consequence of treatment at different times. Our findings indicate that cisplatin mainly contributes to phospholipid biosynthesis while CasIIgly affects processes such as carbohydrates and nucleotides metabolism. Also, we observed that CasIIgly treatment has an important and fast effect over MDA-MB-231 cell metabolism, which makes it a good alternative for treatment in this type of cancer.

NMR-based metabolomics of human cerebrospinal fluid identifies signature of brain death.

INTRODUCTION: Brain death (BD) is the irreversible cessation of all functions of the entire brain, including the brainstem. Cerebrospinal fluid (CSF) is a biological liquid that circulates in brain and spine. Metabolomics is able to reveal the response of biological systems to diverse factors in a specific moment or condition. Therefore, the study of this neurological condition through metabolic profiling using high resolution Nuclear Magnetic Resonance (NMR) spectroscopy is important for understanding biochemical events. OBJECTIVES: The aim of the current study is to identify the metabolomics signature of BD using 1H-NMR spectroscopy in human CSF. METHODS: 1H-NMR spectroscopy has been employed for metabolomic untargeted analysis in 46 CSF samples: 22 control and 24 with BD. Spectral data were further subjected to multivariate analysis. RESULTS: Statistically significant multivariate models separated subject's samples with BD from controls and revealed twenty one discriminatory metabolites. The statistical analysis of control and BD subjects using Orthogonal Projections to Latent Structures Discriminant Analysis (OPLS-DA) model resulted in R2X of 0.733 and Q2 of 0.635. An elevation in the concentration of statistically discriminant metabolites in BD was observed. CONCLUSION: This study identifies a metabolic signature associated with BD and the most relevant enriched selected metabolic pathways.

1H NMR profiling and chemometric analysis as an approach to predict the leishmanicidal activity of dichloromethane extracts from Lantana camara (L.).

The application of 1H NMR spectroscopy and chemometrics for the analysis of extracts of Lantana camara is described. This approach allowed to predict the leishmanicidal activity of samples obtained at different harvest times from their 1H NMR spectra. The anti-leishmanial activity of dichloromethane extracts obtained from the aerial parts of L. camara was measured using an in vitro assay. As the extracts displayed differences in their activity according to a one-way ANOVA analysis, their 1H NMR spectra were subjected to multivariate analysis using exploratory (Principal Component Analysis (PCA) and Anova Simultaneous Component Analysis (ASCA)) and regression, (Partial Least Squares Regression to Latent Structures (PLS)) chemometrics methods. These analyses allowed to establish and characterize a predictive model capable of determining the anti-leishmanial activity of Lantana camara dichloromethane extracts from their 1H NMR spectra. Figures of merit of the developed method are given as well. The identified chemical signals responsible for the iPLS calibration model corresponded to the presence of eicosane, caryophyllene oxide, ß-ionone, tiglic acid, lantanilic acid, camaric acid, and lantadene B; the chemical markers. This study proposed a fast and simple method that avoids the need of using complex biological assays to predict the leishmanicidal activity of L. camara dichloromethane extracts.

Two-photon induced isomerization through a cyaninic molecular antenna in azo compounds.

We present a new design for non-linear optically responsive molecules based on a modular scheme where a polymethinic antenna section with important two-photon absorption properties is bonded to an isomerizable actuator section composed of a stilbenyl-azopyrrole unit. Upon two photon excitation, energy migration from the antenna-localized second singlet excited state to the stilbenyl-azopyrrole section allows for efficient indirect excitation and phototransformation of this actuator.

Synthesis, complete NMR assignment and structural study of a steroidal dimer of 17α-ethynyl-5α,10α-estran-17ß-ol with diethynylbenzene spacer.

A phenylene-bridged steroidal dimer derived from 17α-ethynyl-5α,10α-estran-17ß-ol with molecular rotor-like architecture was synthesized to investigate the supramolecular interactions directing the crystallization of these systems. Structures with varying importance in complementarity between H-bonding and hydrophobic interactions can be observed directing the packing of the obtained crystals, depending on the synthetic stage, though conserving the same space group for both systems. Such behavior clearly shows the versatility achievable using steroids as crystal packing directors. Alongside this structural study, the complete NMR assignment is presented for the dimer, and precursors, in which the steroids present an unconventional and noteworthy A-B ring fusion.

Ultrafast Fluorescence Signals from ß-Dihydronicotinamide Adenine Dinucleotide: Resonant Energy Transfer in the Folded and Unfolded Forms.

ß-Dihydronicotinamide adenine dinucleotide (NADH) plays a critical role in biological redox processes. Inside the cell, NADH can be in a folded or an unfolded conformation, depending on the chemical environment that surrounds it. It is known that selective excitation of adenine in NADH can induce energy transfer events from this nucleotide to the reduced nicotinamide chromophore. From the anticipated time scales, this process must compete with adenine's internal conversion channel, which is known to occur in the sub-picosecond time scale. In this work, we studied the dynamics of the excited states of both chromophores in NADH through the time resolution of the spontaneous emission from both nucleotides. Through these experiments, we extend the knowledge about the competition between the different photophysical channels both in the folded and unfolded states. The study involved the folded and unfolded states of NADH by experiments in water and methanol solutions. Our femtosecond fluorescence results were complemented by the first nuclear magnetic resonance through space magnetization transfer measurements on NADH, which establish the solvent-dependent folded versus unfolded states. We determined the dynamics of the excited states by direct excitation of dihydronicotinamide at 380 nm and adenine at 266 nm. From this, we were able to measure for the folded state, a time constant of 90 fs for energy transfer. Additionally, we determined that even in what is referred to as an unfolded state in methanol, non-negligible excitation transfer events do take place. Spontaneous emission anisotropy measurements were used in order to confirm the presence of a minor energy transfer channel in the methanol solutions where the unfolded state dominates.

Structural Changes in Wormlike Micelles on the Incorporation of Small Photoswitchable Molecules.

Chromophores susceptible to light-induced trans-cis isomerization embedded in cylindrical micelles can modify micelles and their light-responsive performance. A small chromophore (4-(phenylazo)benzoate ion) is embedded in cylindrical micelles made of cetyltrimethylammonium bromide (CTAB) and sodium salicylate (NaSal) in water. The microstructure is examined by scanning electron microscopy (SEM) and nuclear magnetic resonance (NMR). Rheological behavior and the length scales of the micellar network are determined by rheology and microrheology. The chromophore substantially modifies the micelles even without UV irradiation. The larger is the chromophore concentration, the smaller is the micellar length. Additional length scales of the micellar network do not substantially vary even when NaCl is added. Chromophore incorporation also modifies the rheology of the micellar solution, although gradient shear banding is preserved. Viscosity decreases as the chromophore concentration increases, and viscoelastic spectra are modified, but when they are correctly rescaled, they can be superimposed. The addition of the chromophore makes the fluids more Maxwellian, particularly when NaCl is also added. When the chromophore is incorporated into the micelles, there is a response after UV irradiation, although it does not produce a significant rheological change.

Exo-Metabolites of Phaseolus vulgaris-Nodulating Rhizobial Strains.

Rhizobia are able to convert dinitrogen into biologically available forms of nitrogen through their symbiotic association with leguminous plants. This results in plant growth promotion, and also in conferring host resistance to different types of stress. These bacteria can interact with other organisms and survive in a wide range of environments, such as soil, rhizosphere, and inside roots. As most of these processes are molecularly mediated, the aim of this research was to identify and quantify the exo-metabolites produced by Rhizobium etli CFN42, Rhizobium leucaenae CFN299, Rhizobium tropici CIAT899, Rhizobium phaseoli Ch24-10, and Sinorhizobium americanum CFNEI156, by nuclear magnetic resonance (NMR). Bacteria were grown in free-living cultures using minimal medium containing sucrose and glutamate. Interestingly, we found that even when these bacteria belong to the same family (Rhizobiaceae) and all form nitrogen-fixing nodules on Phaseolus vulgaris roots, they exhibited different patterns and concentrations of chemical species produced by them.

The structure of (E)-biformene synthase provides insights into the biosynthesis of bacterial bicyclic labdane-related diterpenoids.

The labdane-related diterpenoids (LRDs) are a large group of natural products with a broad range of biological activities. They are synthesized through two consecutive reactions catalyzed by class II and I diterpene synthases (DTSs). The structural complexity of LRDs mainly depends on the catalytic activity of class I DTSs, which catalyze the formation of bicyclic to pentacyclic LRDs, using as a substrate the catalytic product of class II DTSs. To date, the structural and mechanistic details for the biosynthesis of bicyclic LRDs skeletons catalyzed by class I DTSs remain unclear. This work presents the first X-ray crystal structure of an (E)-biformene synthase, LrdC, from the soil bacterium Streptomyces sp. strain K155. LrdC was identified as a part of an LRD cluster of five genes and was found to be a class I DTS that catalyzes the Mg2+-dependent synthesis of bicyclic LRD (E)-biformene by the dephosphorylation and rearrangement of normal copalyl pyrophosphate (CPP). Structural analysis of LrdC coupled with docking studies suggests that Phe189 prevents cyclization beyond the bicyclic LRD product through a strong stabilization of the allylic carbocation intermediate, while Tyr317 functions as a general base catalyst to deprotonate the CPP substrate. Structural comparisons of LrdC with homology models of bacterial bicyclic LRD-forming enzymes (CldD, RmnD and SclSS), as well as with the crystallographic structure of bacterial tetracyclic LRD ent-kaurene synthase (BjKS), provide further structural insights into the biosynthesis of bacterial LRD natural products.

Chemical Constituents with Leishmanicidal Activity from a Pink-Yellow Cultivar of Lantana camara var. aculeata (L.) Collected in Central Mexico.

Lantana camara (L.) is employed by several ethnical groups to treat numerous diseases. Although there are no ethnomedical reports on its use against leishmaniasis, organic extracts prepared from L. camara were shown to display leishmanicidal activity. In the present study, we carried out a bioassay-guided fractionation of the dichloromethane extract from Mexican L. camara in order to identify the compounds responsible for the leishmanicidal activity. Eighteen chromatographic fractions (FI⁻FXVIII) were evaluated in vitro against Leishmania mexicana and L. amazonensis. FII, FX, FXI, FXV, and FXVI showed significant activity against both Leishmania strains, the most potent of which was FXV. Eicosane (1), squalene (2), ß-ionone (3), caryophyllene oxide (4), ß-caryophyllene (5), hexanoic acid (6), tiglic acid (7), a mixture of lantanilic (8) and camaric (9) acids, and lantadene B (10) were identified and obtained from the active fractions and evaluated for their leishmanicidal activity. The mixture of lantanilic (8) and camaric (9) acids (79%/21%) was the most potent one (half maximal inhibitory concentration (IC50) = 12.02 ± 0.36 µM). This study indicates that this cultivar of L. camara has high potential for the development of phytomedicines or as a source of natural products, which might represent lead compounds for the design of new drugs against leishmaniasis.