Resolution 1H Nuclear Magnetic Resonance Research Articles

The serum metabolomic profiles of atrial fibrillation patients treated with direct oral anticoagulants or vitamin K antagonists

AimsLong-term oral anticoagulation is the primary therapy for preventing ischemic stroke in patients with atrial fibrillation (AF). Different types of oral anticoagulant drugs can have specific effects on the metabolism of patients. Here we characterize, for the first time, the serum metabolomic and lipoproteomic profiles of AF patients treated with anticoagulants: vitamin K antagonists (VKAs) or direct oral anticoagulants (DOACs). Materials and methodsSerum samples of 167 AF patients (median age 78 years, 62 % males, 70 % on DOACs treatment) were analyzed via high resolution 1H nuclear magnetic resonance (NMR) spectroscopy. Data on 25 metabolites and 112 lipoprotein-related fractions were quantified and analyzed with multivariate and univariate statistical approaches. Key findingsOur data provide evidence that patients treated with VKAs and DOACs present significant differences in their profiles: lower levels of alanine and lactate (odds ratio: 1.72 and 1.84), free cholesterol VLDL-4 subfraction (OR: 1.75), triglycerides LDL-1 subfraction (OR: 1.80) and 4 IDL cholesterol fractions (ORs ∼ 1.80), as well as higher levels of HDL cholesterol (OR: 0.48), apolipoprotein A1 (OR: 0.42) and 7 HDL cholesterol fractions/subfractions (ORs: 0.40–0.51) are characteristic of serum profile of patients on DOACs' therapy. SignificanceOur results support the usefulness of NMR-based metabolomics for the description of the effects of oral anticoagulants on AF patient circulating metabolites and lipoproteins. The higher serum levels of HDL cholesterol observed in patients on DOACs could contribute to explaining their reduced cardiovascular risk, suggesting the need of further studies in this direction to fully understand possible clinical implications.

Identification and Quantification of Hydrocarbon Functional Groups in Gasoline Using 1H-NMR Spectroscopy for Property Prediction.

Gasoline is one of the most important distillate fuels obtained from crude refining; it is mainly used as an automotive fuel to propel spark-ignited (SI) engines. It is a complex hydrocarbon fuel that is known to possess several hundred individual molecules of varying sizes and chemical classes. These large numbers of individual molecules can be assembled into a finite set of molecular moieties or functional groups that can independently represent the chemical composition. Identification and quantification of groups enables the prediction of many fuel properties that otherwise may be difficult and expensive to measure experimentally. In the present work, high resolution 1H nuclear magnetic resonance (NMR) spectroscopy, an advanced structure elucidation technique, was employed for the molecular characterization of a gasoline sample in order to analyze the functional groups. The chemical composition of the gasoline sample was then expressed using six hydrocarbon functional groups, as follows: paraffinic groups (CH, CH2 and CH3), naphthenic CH-CH2 groups and aromatic C-CH groups. The obtained functional groups were then used to predict a number of fuel properties, including research octane number (RON), motor octane number (MON), derived cetane number (DCN), threshold sooting index (TSI) and yield sooting index (YSI).

Food protein network formation and gelation induced by conductive or microwave heating: A focus on hen egg white

Heat-induced protein polymerization, gelation and the associated molecular scale changes were investigated during conductive and microwave heating (CH and MH, respectively). Given identical temperature-time profiles, polymerization of bovine serum albumin, ovalbumin, egg white (EW) and wheat gluten protein occurred faster and to a greater extent with CH than with MH. Moreover, lanthionine cross-links were present in some MH but in none of the CH samples. Gelation of EW protein by CH or MH was studied in more detail. Differential scanning calorimetry and fluorescence spectroscopy indicated that EW proteins initially denatured to a larger extent with CH than with MH. Size exclusion high performance liquid chromatography demonstrated that CH resulted in more heavily cross-linked EW protein networks than MH. Homogeneous EW gels were obtained by 15 min CH at 80 °C while 120 min MH at this temperature still resulted in inhomogeneous (with liquid and solid zones) gels. This was affirmed by low resolution 1H nuclear magnetic resonance measurements which indicated more mobile water in MH than in CH samples. However, MH gels had higher firmness than the corresponding CH gels. Thus, MH here led to less covalent cross-linking than CH, but still resulted in firmer gels most likely because its protein network relied heavily on non-covalent interactions. Industrial relevanceThis study highlighted differences between microwave and conventional, conductive, heating of different protein materials. While literature is divided about the origin of differences noted between these different heating modes, it is clear from our data that heating mode, given identical temperature-time profiles, impacts the changes that proteins undergo during heating. The knowledge that microwave heating favors protein unfolding, aggregation and cross-linking to a lower extent than conductive heating can likely be exploited in industrial applications.

Phenanthridine derivatives as promising new anticancer agents: synthesis, biological evaluation and binding studies.

Aim: Phenanthridines are an essential class of nitrogenous heterocycles with extensive applications in medicinal chemistry. The development of efficient and eco-friendly methods for the preparation of chirally pure dihydropyrrolo[1,2-f]phenanthridines (5a-h), and their in vitro evaluation and modeling studies as potential anticancer, antioxidant and DNA cleavage agents is reported. Methodology & results: Compounds 5a-h were prepared through a facile one-pot synthesis and characterized by infrared, high resolution mass spectrometry, 1H and 13C nuclear magnetic resonance. The molecules were subjected to virtual screening and docking analysis against selected human molecular targets. Compound 5g displayed good binding properties as well as significant anticancer and DNA cleavage activity. Conclusion: Compound 5g has been identified as a potential lead candidate for further testing against additional cancer cell lines and animal models in future.

Dereplication of Aporphine Alkaloids by UHPLC-HR-ESI-MS/MS and NMR from Duguetia lanceolata St.-Hil (Annonaceae) and Antiparasitic Activity Evaluation

Although the genus Duguetia is well known for producing alkaloids as chemical constituents, there are no reports of alkaloids identified in the species D. lanceolata. Thus, aiming to identify the chemical composition of this species, the dereplication of alkaloidic phase was performed by use of ultra-high performance liquid chromatography high resolution electrospray ionization tandem mass spectrometry (UHPLC-HR-ESI-MS/MS) and 1H nuclear magnetic resonance (NMR). The chromatographic fractionation of the alkaloid extract from Duguetia lanceolata (Annonaceae) leaves afforded four fractions (I-IV) that were shown to be composed of aporphine alkaloids. 1H NMR analysis and UHPLC-HR-ESI-MS/MS based dereplication allowed the identification of eight alkaloids: glaucine (1), norglaucine (2), isocorydine (3), N-methyllaurotetanine (4), oxoglaucine (5), liriodenine (6), lanuginosine (7), dehydroglaucine (8). Compounds 2,3, 4,6 and 7 were described for the first time in this species, while alkaloids 1,5 and 8 are newly discovered in the genus Duguetia. Additionally, the antiparasitic activity of the four fractions was evaluated in vitro against Leishmania infantum and Trypanosoma cruzi. Fraction I, composed exclusively by 1, displayed activity against Leishmania infantum and Trypanosoma cruzi intracellular amastigotes, with half maximal inhibitory concentration (IC50) values of 7.5 and 28.6 µg mL-1, respectively. Fraction IV (constituted by 2,3 and 4) showed activity against promastigotes of Leishmania infantum with IC50 value of 50.0 µg mL-1, while fraction II (constituted by 5 and 6) showed activity against trypomastigotes of Trypanosoma cruzi, with IC50 values of 83.0 µg mL-1. This work showed that fragmentation in UHPLC-HR-ESI-MS/MS combined with 1H NMR analysis of fractions is useful for identifying alkaloids in mixtures. Additionally, it was also demonstrated the potential of aporphine alkaloids from Duguetia lanceolata St. -Hil (Annonaceae) in the search for new drug candidates for neglected diseases.

Synthesis, chemical characterization and antimicrobial activity of new acylhydrazones derived from carbohydrates

A new series of glycosylated acylhydrazones was synthesized and all the chemical structures were confirmed by High Resolution Mass Spectrometry (HRMS), 1H and 13C Nuclear Magnetic Resonance (1H-NMR; 13C-NMR) and Fourier Transform Infrared (FTIR) spectroscopy methods. The mass accuracy between the calculated and found values observed in HRMS analyses were near or lower than 5 ppm, which are acceptable for proposing a molecular formula using this technique. All of the synthesized compounds were screened for their antibacterial, antifungal and antiviral activities. Five compounds (12, 13, 14, 16 and 19) exerted a modest antifungal activity against the strains evaluated. Derivative 14 showed fungicidal activity against Candida glabrata at 173.8 μM and saccharide unit contributed to the increase of the antifungal potential against this strain. New chemical manipulation of derivative 14 can make it possible to obtain new potentially antimicrobial agents.

A minimalist functional group (MFG) approach for surrogate fuel formulation

Surrogate fuel formulation has drawn significant interest due to its relevance towards understanding combustion properties of complex fuel mixtures. In this work, we present a novel approach for surrogate fuel formulation by matching target fuel functional groups, while minimizing the number of surrogate species. Five key functional groups; paraffinic CH3, paraffinic CH2, paraffinic CH, naphthenic CHCH2 and aromatic CCH groups in addition to structural information provided by the Branching Index (BI) were chosen as matching targets. Surrogates were developed for six FACE (Fuels for Advanced Combustion Engines) gasoline target fuels, namely FACE A, C, F, G, I and J. The five functional groups present in the fuels were qualitatively and quantitatively identified using high resolution 1H Nuclear Magnetic Resonance (NMR) spectroscopy. A further constraint was imposed in limiting the number of surrogate components to a maximum of two. This simplifies the process of surrogate formulation, facilitates surrogate testing, and significantly reduces the size and time involved in developing chemical kinetic models by reducing the number of thermochemical and kinetic parameters requiring estimation. Fewer species also reduces the computational expenses involved in simulating combustion in practical devices. The proposed surrogate formulation methodology is denoted as the Minimalist Functional Group (MFG) approach. The MFG surrogates were experimentally tested against their target fuels using Ignition Delay Times (IDT) measured in an Ignition Quality Tester (IQT), as specified by the standard ASTM D6890 methodology, and in a Rapid Compression Machine (RCM). Threshold Sooting Index (TSI) and Smoke Point (SP) measurements were also performed to determine the sooting propensities of the surrogates and target fuels. The results showed that MFG surrogates were able to reproduce the aforementioned combustion properties of the target FACE gasolines across a wide range of conditions. The present MFG approach supports existing literature demonstrating that key functional groups are responsible for the occurrence of complex combustion properties. The functional group approach offers a method of understanding the combustion properties of complex mixtures in a manner which is independent, yet complementary, to detailed chemical kinetic models. The MFG approach may be readily extended to formulate surrogates for other complex fuels.

Sodium pyruvate protect occipital cortex of rats with repetitive and severe neonatal hypoglycemia detected by high resolution 1H nuclear magnetic resonance spectroscopy

Objectives To investigate the occipital cortex metabolite alterations in repetitive and severe neonatal hypoglycemia rats treated with sodium pyruvate and to reveal the protective role of sodium pyruvate using high resolution 1H nuclear magnetic resonance spectroscopy. Methods Thirty-six 2-day-old Sprague-Dawley rats were randomly divided into hypoglycemia group and pyruvate group with 18 rats in each group. Rats in both groups received intraperitoneal injections of insulin (40 U/kg body weight) at 2, 4 and 6 days of age to induce severe hypoglycemia (blood glucose value ≤1.4 mmol/L). In the hypoglycemia group, 2.5 hours after insulin injection, intraperitoneal injection of 50% glucose (2 ml/kg) was administered to terminate hypoglycemia, while in the pyruvate group, 50% glucose (2 ml/kg) and sodium pyruvate solution 2.5 ml/kg (500 mg/kg) were injected. Terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay was used to observe the status of injured neurons in six neonatal rats, and metabolite changes in occipital cortex of the other 12 rats were detected by 1H nuclear magnetic resonance spectroscopy. The difference between the two groups was compared by independent-samples t test. Results Neonatal rats of both groups reached severe hypoglycemia level 2.5 hours after insulin injection. Compared with hypoglycemia group, pyruvate group had fewer injured neurons (45±5 vs 113±12, t=0.782, P=0.013) and lower injured index in the occipital cortex (0.15±0.03 vs 0.36±0.06, t=1.143, P=0.020). Pyruvate group showed significant decreases in the concentration of taurine [(13.31±2.06) vs (18.44±3.86) mol/kg, t=8.231], glutamine[(1.50±0.24) vs (2.02±0.40) mol/kg, t=3.137], glutamate[(7.04±0.95) vs (9.40±1.73) mol/kg, t=6.449], aspartate[(1.51±0.28) vs (2.15±0.58) mol/kg, t=2.561] and creatine [(6.37±0.99) vs (8.46±1.77) mol/kg, t=4.226] in the occipital cortex (all P'<0.017). Conclusions Simultaneous use of glucose and sodium pyruvate to terminate hypoglycemia in repetitive and severe neonatal hypoglycemia rats can effectively alleviate severe hypoglycemia-induced occipital lobe damage via regulating excitatory amino acid neurotransmitters, energy metabolism and other metabolic pathways. Key words: Hypoglycemia; Pyruvates; Occipital lobe; Magnetic resonance spectroscopy; Animals, newborn

Non-invasive diagnosis of papillary thyroid microcarcinoma: a NMR-based metabolomics approach.

Papillary thyroid microcarcinoma (PTMC) is a subtype of papillary thyroid carcinoma (PTC). Because its diameter is less than 10 mm, diagnosing it accurately is difficult with traditional methods such as image examinations and FNA (Fine Needle Aspiration). Investigating the metabolic changes induced by PTMC may enhance the understanding of its pathogenesis and provide important information for a new diagnosis method and treatment plan. In this study, high resolution magic angle spin (HRMAS) spectroscopy and 1H-nuclear magnetic resonance (1H-NMR) spectroscopy were used to screen metabolic changes in thyroid tissues and plasma from PTMC patients respectively. The results revealed reduced levels of fatty acids and elevated levels of several amino acids (phenylalanine, tyrosine, lactate, serine, cystine, lysine, glutamine/glutamate, taurine, leucine, alanine, isoleucine and valine) in thyroid tissues, as well as reduced levels of amino acids such as valine, tyrosine, proline, lysine, leucine and elevated levels of glucose, mannose, pyruvate and 3-hydroxybutyrate in plasma, are involved in the metabolic alterations in PTMC. In addition, a receiver operating characteristic (ROC) curve model for PTMC prediction was able to classify cases with good sensitivity and specificity using 9 significant changed metabolites in plasma. This work illustrates that the NMR-based metabolomics approach is capable of providing more sensitive diagnostic results and more systematic therapeutic information for PTMC.

TG/DTG, FT-ICR Mass Spectrometry, and NMR Spectroscopy Study of Heavy Fuel Oil

There is an increasing interest in the comprehensive study of heavy fuel oil (HFO) due to its growing use in furnaces, boilers, marines, and recently in gas turbines. In this work, the thermal combustion characteristics and chemical composition of HFO were investigated using a range of techniques. Thermogravimetric analysis (TGA) was conducted to study the nonisothermal HFO combustion behavior. Chemical characterization of HFO was accomplished using various standard methods in addition to direct infusion atmospheric pressure chemical ionization Fourier transform ion cyclotron resonance mass spectrometry (APCI-FTICR MS), high resolution 1H nuclear magnetic resonance (NMR), 13C NMR, and two-dimensional heteronuclear multiple bond correlation (HMBC) spectroscopy. By analyzing thermogravimetry and differential thermogravimetry (TG/DTG) results, three different reaction regions were identified in the combustion of HFO with air, specifically, low temperature oxidation region (LTO), fuel deposition (FD), and hig...

Antigiardial activity of novel triazolyl-quinolone-based chalcone derivatives: when oxygen makes the difference

Giardiasis is a common diarrheal disease worldwide caused by the protozoan parasite Giardia intestinalis. It is urgent to develop novel drugs to treat giardiasis, due to increasing clinical resistance to the gold standard drug metronidazole (MTZ). New potential antiparasitic compounds are usually tested for their killing efficacy against G. intestinalis under anaerobic conditions, in which MTZ is maximally effective. On the other hand, though commonly regarded as an ‘anaerobic pathogen,’ G. intestinalis is exposed to relatively high O2 levels in vivo, living attached to the mucosa of the proximal small intestine. It is thus important to test the effect of O2 when searching for novel potential antigiardial agents, as outlined in a previous study [Bahadur et al. (2014) Antimicrob. Agents Chemother. 58, 543]. Here, 45 novel chalcone derivatives with triazolyl-quinolone scaffold were synthesized, purified, and characterized by high resolution mass spectrometry, 1H and 13C nuclear magnetic resonance and infrared spectroscopy. Efficacy of the compounds against G. intestinalis trophozoites was tested under both anaerobic and microaerobic conditions, and selectivity was assessed in a counter-screen on human epithelial colorectal adenocarcinoma cells. MTZ was used as a positive control in the assays. All the tested compounds proved to be more effective against the parasite in the presence of O2, with the exception of MTZ that was less effective. Under anaerobiosis eighteen compounds were found to be as effective as MTZ or more (up to three to fourfold); the same compounds proved to be up to >100-fold more effective than MTZ under microaerobic conditions. Four of them represent potential candidates for the design of novel antigiardial drugs, being highly selective against Giardia trophozoites. This study further underlines the importance of taking O2 into account when testing novel potential antigiardial compounds.

Metabolic Profiling of Human Colorectal Cancer Using High Resolution 1H Nuclear Magnetic Resonance Spectroscopy

AbstractColorectal cancer (CRC) is the third commonest malignancy cancer worldwide. Clear understandings of global metabolic profiling of the normal mucosa and cancer tissues are vitally important to aid optimizing the clinical management strategy and understanding CRC biology. We studied metabolic characteristics of 20 CRC and 20 distant normal mucosa tissues extracts from 20 patients using high resolution 1H NMR spectroscopy in conjunction with multivariate analyses, such as principal component analysis (PCA). Compared with distant normal mucosa tissues, lactate, taurine, ornithine and polyamine were present at significantly higher levels in CRC tissue extracts whereas myo‐inositol was present at significantly lower level. Two metabolites ratios such as myo‐inositol/taurine and myo‐inositol/(ornithine+polyamine) appear to be the most valuable biomarkers for the differentiation CRC from normal mucosa tissues. Our data suggested that HR 1H NMR spectroscopy combined with multivariate analyses is a potentially useful technology for detecting malignant changes in the normal mucosa tissues, the technique may be further exploited for future CRC biomarker research or identification of targets for therapeutic manipulations.

P1.45 Application of NMR spectroscopy in the study of mdx mouse

The mdx mouse is the classical animal model for Duchenne Muscular dystrophy, showing similar molecular and protein defects. It is widely used to investigate the pathogenesis of the disease aiming the development of therapeutic strategies. The mdx, however, does not show the significant muscle weakness observed in humans, and the diaphragm muscle is usually used as a marker of progression, due to its severe pattern of degeneration. High resolution 1H nuclear magnetic resonance spectroscopy (MRS) was used in this work to study the metabolic profile of quadriceps and diaphragm muscles, comparing mdx and control mice at 3 and 6months of age.

Antidiabetic efficacy of BRL 49653, a potent orally active insulin sensitizing agent, assessed in the C57BL/KsJ db/db diabetic mouse by non-invasive 1H NMR studies of urine.

High resolution 1H nuclear magnetic resonance (NMR) spectroscopic analysis of biofluids is a recently established tool for evaluating inherited and acquired errors in metabolic control. In the present study 1H NMR analysis of urine was used to monitor efficacy of BRL 49653, a potent and selective antihyperglycaemic agent, following oral administration for up to 36 weeks to the genetically diabetic C57BL/KsJ db/db mouse. The effects of BRL 49653 on carbohydrate and fatty acid metabolism were monitored by determination of changes in concentrations of low molecular weight urinary metabolites. A qualitative comparison of the NMR spectra of urine from untreated diabetic mice with those of lean littermates and literature examples revealed several abnormalities, the majority of which could be explained in terms of the non-insulin dependent diabetes syndrome exhibited by these animals. Quantitatively the most prominent was the extreme glycosuria of both young (8-12 weeks; 0.9 g glucose kg-1 h-1) and older (42 weeks; 2 g glucose kg-1 h-1) diabetic mice. This was accompanied by the excretion of a number of unassigned sugar derivatives and by ketone bodies. Administration of BRL 49653 (3 mumol kg-1) to db/db mice for 24 days reduced blood glucose concentrations to values comparable with non-diabetic lean littermates and reduced glycosuria by > 90%. BRL 49653 significantly reduced excretion of unassigned sugars, acetate, lactate, and the ketone bodies, acetoacetate, 3-D-hydroxybutyrate and acetone. The anti-diabetic efficacy of BRL 49653, assessed from the pattern of urinary metabolites, was maintained over a 36-week treatment period. These results demonstrate the value of 1H NMR to evaluate non-invasively the efficacy of novel therapeutic agents.

Metabolomic Changes Accompanying Transformation and Acquisition of Metastatic Potential in a Syngeneic Mouse Mammary Tumor Model

Breast cancer is the most common cancer type for women in the western world. Despite decades of research, the molecular processes associated with breast cancer progression are still inadequately defined. Here, we focus on the systematic alteration of metabolism by using the state of the art metabolomic profiling techniques to investigate the changes of 157 metabolites during the progression of normal mouse mammary epithelial cells to an isogenic series of mammary tumor cell lines with increasing metastatic potentials. Our results suggest a two-step metabolic progression hypothesis during the acquisition of tumorigenic and metastatic abilities. Metabolite changes accompanying tumor progression are identified in the intracellular and secreted forms in several pathways, including glycolysis, the tricarboxylic acid cycle, the pentose phosphate pathway, fatty acid and nucleotide biosynthesis, and the GSH-dependent antioxidative pathway. These results suggest possible biomarkers of breast cancer progression as well as opportunities of interrupting tumor progression through the targeting of metabolic pathways.

Detection and quantification of phenolic compounds in olive oil by high resolution 1H nuclear magnetic resonance spectroscopy

High resolution 1H NMR spectroscopy has been employed as a versatile and rapid method to analyze the polar fraction of extra virgin olive oils containing various classes of phenolic compounds. The strategy for identification of phenolic compounds is based on the NMR chemical shifts of a large number of model compounds assigned by using two-dimensional (2D) NMR spectroscopy. Furthermore, 2D NMR was applied to phenolic extracts in an attempt to discover additional phenolic compounds. The 1H NMR methodology was successful in detecting simple phenols, such as p-coumaric acid, vanillic acid, homovanillyl alcohol, vanillin, free tyrosol, and free hydroxytyrosol, the flavonols apigenin and luteolin, the lignans (+) pinoresinol, (+) 1-acetoxypinoresinol and syringaresinol, two isomers of the aldehydic form of oleuropein and ligstroside, the dialdehydic form of oleuropein and ligstroside lacking a carboxymethyl group, and finally total hydroxytyrosol and total tyrosol reflecting the total amounts of free and esterified hydroxytyrol and tyrosol, respectively. The absolute amount of each phenolic constituent was determined in the polar fraction by using anhydrous 1,3,5-triazine as an internal standard.

Calcium silicate hydrates investigated by solid‐state high resolution 1H and 29Si nuclear magnetic resonance

This work focuses on phases formed during cement hydration under high pressure and temperature: portlandite Ca(OH) 2 (CH); hillebrandite Ca 2(SiO 3)(OH) 2 (β‐dicalcium silicate hydrate); calcium silicate hydrate (C–S–H); jaffeite Ca 6(Si 2O 7)(OH) 6 (tricalcium silicate hydrate); α‐C 2SH Ca 2(SiO 3)(OH) 2 (α‐dicalcium silicate hydrate); xonotlite Ca 6(Si 6O 17)(OH) 2 and kilchoanite Ca 6(SiO 4)(Si 3O 10). Portlandite and hillebrandite were synthesized and characterised by high resolution solid‐state 1H and 29Si Nuclear Magnetic Resonance. In addition, information from the literature concerning the last five phases was gathered. In certain cases, a schematic 3D‐structure could be determined. These data allow identification of the other phases present in a mixture. Their morphology was also observed by Scanning Electron Microscopy.

System level metabolic effects of a Schistosoma japonicum infection in the Syrian hamster

A metabolic profiling strategy was used to investigate the metabolic responses of Syrian hamsters (SLAC) to a Schistosoma japonicum infection using high resolution 1H nuclear magnetic resonance (NMR) spectroscopy and pattern recognition. In two independent experiments, male hamsters were each infected with 100 S. japonicum cercariae. At days 34–36 post-infection, urine was obtained from hamsters housed individually in metabolism cages. At the same time, urine was collected from age- and sex-matched infection-free control hamsters. The main biochemical effects of a S. japonicum infection in the hamster consisted of reduced levels of urinary tricarboxylic acid cycle intermediates, including citrate and succinate and increased levels of pyruvate. In addition, a range of microbial-related metabolites, such as hippurate, p-cresol glucuronide, phenylacetylglycine and trimethylamine were also associated with a S. japonicum infection. Most of the observed biochemical effects were in common with those previously characterized for a S. mansoni infection in a mouse host. The major distinguishing consequence of a S. japonicum infection in the hamster was the inhibition of manufacture or utilization of short-chain fatty acids, when compared to a S. mansoni infection in the mouse.

High resolution 1H nuclear magnetic resonance in the study of edible oils and fats

In this paper a review of the applications of high resolution 1H NMR to the study of edible oils and fats is reported. Aspects such as sample preparation and acquisition parameters are treated. The assignment of the 1H NMR signals of the major and minor components of unoxidized and of thermally stressed edible oils and fats is given. The usefulness of this technique in the determination of degree of unsaturation and of the proportion of the different acyl groups, is examined. In addition, the capability of this technique to assess the quality and genuineness of olive oil and to detect adulterations is discussed. Finally, its contribution in the study of the oxidation of edible oils and fats and its effectiveness to determine not only primary but also secondary oxidation products, and also oxidative stability is shown.

Comparative biochemistry and short-term starvation effects on the earthworms Eisenia veneta and Lumbricus terrestris studied by 1H NMR spectroscopy and pattern recognition

The aim of this work was to determine the applicability of high resolution 1H nuclear magnetic resonance (NMR) spectroscopic methods to the analysis of earthworm extracts, as a prelude to toxicity testing. This has been applied to the characterisation and examination of starvation effects on the endogenous metabolites of extracts of Eisenia veneta and Lumbricus terrestris in Ringer solution. Identified metabolites include free amino acids, sugars and low molecular weight organic acids. In both E. veneta and L. terrestris NMR signals from: alanine, arginine, asparagine, glutamate, glycine, isoleucine, leucine, phenylalanine, threonine, tryptophan, tyrosine, valine, glucose, citrate, fumarate, inosine, lactate, propanone, succinate and uridine were detected. The E. veneta extracts contained N-methyl-nicotinic and urocanic acids that were not detected in L. terrestris. Higher concentrations of inosine, lysine and malonic acid were also observed for E. veneta. In L. terrestris a higher amount of glucose was observed. The effect of short-term starvation on E. veneta showed small metabolic changes for the first 5 days, with clear differences at 6 and 7 days. These included increases in glutamate, citrate, aspartate and isoleucine, and decreases in lysine, isoleucine and threonine. In contrast L. terrestris showed no consistent endogenous low molecular weight metabolite changes as detected by 1H NMR spectroscopy over this period, suggesting this species to be more suitable for soil toxicity testing and monitoring using this procedure.