PubMed

Rapid and noninvasive technique to assess the metabolomics profile of bovine embryos produced in vitro by Raman spectroscopy. Biomed Opt Express. 2015 Aug 1;6(8):2830-2839 Authors: Santos EC, Martinho HS, Annes K, Leite RF, Milazzotto MP Abstract Morphological assessments are used to select embryos with the highest implantation potential, however it is still very limited. The development of new technologies, such as Raman spectroscopy have improved quantitative and qualitative analysis, and consequently led to a better characterization of embryos and improvements on the prediction of their potential. Therefore, we propose a method based on the conventional in vitro culture system of bovine embryos, and the subsequent analysis of the culture media drops by Raman spectroscopy. Our results obtained by PCA analysis clearly showed a separation of the spectral profiles from culture media drops with and without embryos. PMID: 26309747 [PubMed - as supplied by publisher]

Alterations of plasma metabolite profiles related to adipose tissue distribution and cardiometabolic risk. Am J Physiol Endocrinol Metab. 2015 Aug 25;:ajpendo.00231.2015 Authors: Boulet MM, Chevrier G, Grenier-Larouche T, Pelletier M, Nadeau M, Scarpa J, Prehn C, Marette A, Adamski J, Tchernof A Abstract CONTEXT: Metabolomic profiling of obese individuals revealed altered concentrations of many metabolites, especially branched-chain amino acids (BCAA), possibly linked to altered adipose tissue BCAA catabolism. OBJECTIVE: We tested the hypothesis that some features of this metabolite signature relate closely to visceral obesity and concomitant alterations in cardiometabolic risk factors. We also postulated that alterations in BCAA-catabolizing enzymes are predominant in visceral adipose tissue. METHODS: Fifty-nine women (BMI 20-41 kg/m(2)) undergoing gynecologic surgery were recruited and characterized for overall and regional adiposity, blood metabolite levels using targeted metabolomics and cardiometabolic risk factors. Adipose samples (visceral and subcutaneous) were obtained and used for gene expression and western blot analyses. RESULTS: Obese women had significantly higher circulating BCAA and Kynurenine/Tryptophan (KYN/Trp) ratio than lean or overweight women (p<0.01). Principal component analysis confirmed that factors related to AA and the KYN/Trp ratio were positively associated with BMI, fat mass, visceral or subcutaneous adipose tissue area and subcutaneous adipocyte size (p≤0.05). AA-related factor was positively associated with HOMA-IR (p≤0.01). Factors reflecting glycerophospholipids and sphingolipids levels were mostly associated with altered blood lipid concentrations (p≤0.05). Glutamate level was the strongest independent predictor of visceral adipose tissue area (r=0.46, p<0.001). Obese women had lower expression and protein levels of BCAA-catabolizing enzymes in visceral adipose tissue compared to overweight or lean women (p≤0.05). CONCLUSIONS: Among metabolites altered in obesity, plasma concentrations of BCAA and the KYN/Trp ratio are closely related to increased adiposity. Alterations in expression and protein levels of BCAA-catabolizing enzymes are predominant in visceral adipose tissue. PMID: 26306599 [PubMed - as supplied by publisher]

Related Articles Low- and high-grade bladder cancer appraisal via serum-based proteomics approach. Clin Chim Acta. 2014 Sep 25;436:97-103 Authors: Bansal N, Gupta A, Sankhwar SN, Mahdi AA Abstract OBJECTIVE: To address the shortcomings of urine cytology and cystoscopy for screening and grading of urinary bladder cancer (BC) we applied a serum-based proteomics approach as a surrogate tactic for rapid BC probing. METHODS: This study was performed on 90 sera samples comprising of low-grade (LG, n=33) and high-grade (HG, n=32) BC, and healthy controls (HC, n=25). Two-dimensional gel electrophoresis (2DE) tactic was executed to describe serum proteome. MALDI-TOF-MS (MS) was used to identify the characteristics of aberrantly expressed proteins in 2DE and validated using Western blot (WB) and ELISA approach. Receiver operating characteristics (ROC) curve analysis was also performed to determine the clinical usefulness of these proteins to discriminate among LG, HG and HC cohorts. RESULTS: This comprehensive approach of 2DE, MS, WB and ELISA reveals five differentially expressed proteins. Among them two biomarkers (S100A8 and S100A9) were able to accurately (ROC, 0.946) distinguish 81% of BC (LG+HG) cases compared to HC with highest sensitivity and specificity. With a comparable tactic, two biomarkers (S100A8 and S100A4) were able to precisely (ROC, 0.941) discriminate 92% of LG cases from HG with utmost sensitivity and specificity. CONCLUSIONS: Serum proteomics probing appears to be an encouraging and least-invasive tactic for screening and grading of BC. PMID: 24875752 [PubMed - indexed for MEDLINE]

Oligoribonuclease is a central feature of cyclic diguanylate signaling in Pseudomonas aeruginosa. Proc Natl Acad Sci U S A. 2015 Aug 24; Authors: Cohen D, Mechold U, Nevenzal H, Yarmiyhu Y, Randall TE, Bay DC, Rich JD, Parsek MR, Kaever V, Harrison JJ, Banin E Abstract The second messenger cyclic diguanylate (c-di-GMP) controls diverse cellular processes among bacteria. Diguanylate cyclases synthesize c-di-GMP, whereas it is degraded by c-di-GMP-specific phosphodiesterases (PDEs). Nearly 80% of these PDEs are predicted to depend on the catalytic function of glutamate-alanine-leucine (EAL) domains, which hydrolyze a single phosphodiester group in c-di-GMP to produce 5'-phosphoguanylyl-(3',5')-guanosine (pGpG). However, to degrade pGpG and prevent its accumulation, bacterial cells require an additional nuclease, the identity of which remains unknown. Here we identify oligoribonuclease (Orn)-a 3'→5' exonuclease highly conserved among Actinobacteria, Beta-, Delta- and Gammaproteobacteria-as the primary enzyme responsible for pGpG degradation in Pseudomonas aeruginosa cells. We found that a P. aeruginosa Δorn mutant had high intracellular c-di-GMP levels, causing this strain to overexpress extracellular polymers and overproduce biofilm. Although recombinant Orn degraded small RNAs in vitro, this enzyme had a proclivity for degrading RNA oligomers comprised of two to five nucleotides (nanoRNAs), including pGpG. Corresponding with this activity, Δorn cells possessed highly elevated pGpG levels. We found that pGpG reduced the rate of c-di-GMP degradation in cell lysates and inhibited the activity of EAL-dependent PDEs (PA2133, PvrR, and purified recombinant RocR) from P. aeruginosa. This pGpG-dependent inhibition was alleviated by the addition of Orn. These data suggest that elevated levels of pGpG exert product inhibition on EAL-dependent PDEs, thereby increasing intracellular c-di-GMP in Δorn cells. Thus, we propose that Orn provides homeostatic control of intracellular pGpG under native physiological conditions and that this activity is fundamental to c-di-GMP signal transduction. PMID: 26305928 [PubMed - as supplied by publisher]

Metabolic footprint of epiphytic bacteria on Arabidopsis thaliana leaves. ISME J. 2015 Aug 25; Authors: Ryffel F, Helfrich EJ, Kiefer P, Peyriga L, Portais JC, Piel J, Vorholt JA Abstract The phyllosphere, which is defined as the parts of terrestrial plants above the ground, is a large habitat for different microorganisms that show a high extent of adaption to their environment. A number of hypotheses were generated by culture-independent functional genomics studies to explain the competitiveness of specialized bacteria in the phyllosphere. In contrast, in situ data at the metabolome level as a function of bacterial colonization are lacking. Here, we aimed to obtain new insights into the metabolic interplay between host and epiphytes upon colonization of Arabidopsis thaliana leaves in a controlled laboratory setting using environmental metabolomics approaches. Quantitative nuclear magnetic resonance (NMR) and imaging high-resolution mass spectrometry (IMS) methods were used to identify Arabidopsis leaf surface compounds and their possible involvement in the epiphytic lifestyle by relative changes in compound pools. The dominant carbohydrates on the leaf surfaces were sucrose, fructose and glucose. These sugars were significantly and specifically altered after epiphytic leaf colonization by the organoheterotroph Sphingomonas melonis or the phytopathogen Pseudomonas syringae pv. tomato, but only to a minor extent by the methylotroph Methylobacterium extorquens. In addition to carbohydrates, IMS revealed surprising alterations in arginine metabolism and phytoalexin biosynthesis that were dependent on the presence of bacteria, which might reflect the consequences of bacterial activity and the recognition of not only pathogens but also commensals by the plant. These results highlight the power of environmental metabolomics to aid in elucidating the molecular basis underlying plant-epiphyte interactions in situ. PMID: 26305156 [PubMed - as supplied by publisher]

Evolution of metabolomics profile of crab paste during fermentation. Food Chem. 2016 Feb 1;192:886-92 Authors: Chen D, Ye Y, Chen J, Yan X Abstract Crab paste is regularly consumed by people in the coastal area of China. The fermentation time plays a key role on the quality of crab paste. Here, we investigated the dynamic evolution of metabolite profile of crab paste during fermentation by combined use of NMR spectroscopy and multivariate data analysis. Our results showed that crab paste quality was significantly affected by fermentation. The quality change was manifested in the decline of lactate, betaine, taurine, trimethylamine-N-oxide, trigonelline, inosine, adenosine diphosphate, and 2-pyridinemethanol, and in the fluctuation of a range of amino acids as well as in the accumulation of glutamate, sucrose, formate, acetate, trimethylamine, and hypoxanthine. Trimethylamine production and its increased level with fermentation could be considered as a freshness index of crab paste. These results contribute to quality assessment of crab paste and confirm the metabolomics technique as a useful tool to provide important information on the crab paste quality. PMID: 26304425 [PubMed - in process]

Metabolomics and microbiological profile of Italian mozzarella cheese produced with buffalo and cow milk. Food Chem. 2016 Feb 1;192:618-24 Authors: Pisano MB, Scano P, Murgia A, Cosentino S, Caboni P Abstract Italian buffalo mozzarella (BM) cheese metabolite profile and microbial communities were characterised and compared to cow mozzarella (CM). Polar metabolite profiles were studied by gas-chromatography mass-spectrometry (GC-MS) and results elaborated by multivariate analysis (MVA). BM produced using natural whey starter cultures (NWS) exhibited a higher microbial diversity with less psychrotrophic bacteria. BM samples were higher in threonine, serine, valine, and lower in orotic acid and urea. CM produced with commercial starters (CMS) had the highest count of Streptococcus thermophilus and higher levels of galactose and phenylalanine. CM obtained by direct acidification (CMA) had lower microbial counts and higher levels of urea and sugars. Orotic acid was the only metabolite linked to milk animal origin. Results indicated that this metabolite pool well reflects the different production protocols and microbial complexity of these dairy products. This approach can help to protect the designation of origin of Italian buffalo mozzarella. PMID: 26304391 [PubMed - in process]

Global metabolite profiling and diagnostic ion filtering strategy by LC-QTOF MS for rapid identification of raw and processed pieces of Rheum palmatum L. Food Chem. 2016 Feb 1;192:531-40 Authors: Liu Y, Li L, Xiao YQ, Yao JQ, Li PY, Yu DR, Ma YL Abstract Due to its variety of functions, rhubarb has been used for thousands of years in many countries. It is commonly used after processing. Processing usually affect the chemical profile and the contents of active compounds in herbals, leading to changes of their bioactivities. Here, an approach of metabolite profiling and diagnostic ion filtering strategy with liquid chromatography-quadrupole/time-of-flight mass spectrometry was established for rapid identification of raw and processed pieces of Rheum palmatum L. (RPL). The comprehensive and unbiased information of 30 batches of RPL covering raw and two general processing methods were given by metabolomic profiles. Using molecular feature extraction algorithm, non-targeted compounds were analyzed in minutes. In total, 73 characteristic markers were extracted and identified by diagnostic ion filtering. They have been further analyzed by partial least squares-support vector machine-based pattern recognition. The comprehensive and rapid method for raw and processed pieces of RPL classification shows good sensitivity, specificity and prediction performance. PMID: 26304381 [PubMed - in process]

Sulfur dioxide residue in sulfur-fumigated edible herbs: The fewer, the safer? Food Chem. 2016 Feb 1;192:119-24 Authors: Duan SM, Xu J, Bai YJ, Ding Y, Kong M, Liu HH, Li XY, Zhang QS, Chen HB, Liu LF, Li SL Abstract The residual content of sulfur dioxide is frequently regarded as the exclusive indicator in the safety evaluation of sulfur-fumigated edible herbs. To examine the feasibility of such assessment criteria, here the variations in residual sulfur dioxide content during sulfur-fumigation and the potential mechanisms involved were investigated, using Angelicae Sinensis Radix (ASR) as a model herb. The residual sulfur dioxide content and ten major bioactive components in sulfur-fumigated ASR samples were dynamically examined at 13 successive time points within 72h sulfur-fumigation. The relationship between the content variation tendency of sulfur dioxide and the ten chemicals was discussed. The results suggested that sulfur dioxide-involved chemical transformation of the original components in ASR might cause large consumption of residual sulfur dioxide during sulfur-fumigation. It implies that without considering the induced chemical transformation of bioactive components, the residual sulfur dioxide content alone might be inadequate to comprehensively evaluate the safety of sulfur-fumigated herbs. PMID: 26304328 [PubMed - in process]

Approaches for predicting effects of unintended environmental exposure to an endocrine active pharmaceutical, tamoxifen. Environ Toxicol. 2015 Aug 25; Authors: Mills LJ, Henderson WM, Jayaraman S, Gutjahr-Gobell RE, Zaroogian GE, Horowitz DB, Laws SC Abstract Tamoxifen is an endocrine-active pharmaceutical (EAP) that is used world-wide. Because tamoxifen is a ubiquitous pharmaceutical and interacts with estrogen receptors, a case study was conducted with this compound to (1) determine effects on reproductive endpoints in a nontarget species (i.e., a fish), (2) compare biologically-active metabolites across species, (3) assess whether in vitro assays predict in vivo results, and (4) investigate metabolomic profiles in tamoxifen-treated fish to better understand the biological mechanisms of tamoxifen toxicity. In reproductive assays, tamoxifen exposure caused a significant reduction in egg production and significantly increased ovarian aromatase activity in spawning adult cunner fish (Tautogolabrus adspersus). In plasma from tamoxifen-exposed cunner, the predominant metabolite was 4-hydroxytamoxifen, while in rats it was N-desmethyltamoxifen. Because 4-hydroxytamoxifen is a more biologically active metabolite than N-desmethyltamoxifen, this difference could result in a different level of risk for the two species. The results of in vitro assays with fish hepatic microsomes to assess tamoxifen metabolism did not match in vivo results, indicating probable differences in excretion of tamoxifen metabolites in fish compared with rats. For the first time, a complete in vitro characterization of the metabolism of tamoxifen using fish microsomes is presented. Furthermore, a metabolomic investigation of cunner gonad extracts demonstrates that tamoxifen alters the biochemical profile in this nontarget species. Understanding the consequence of tamoxifen exposure in nontarget species, and assessing the discrepancies between sex- and species-mediated endpoints, is a step toward understanding how to accurately assess the risks posed by EAPs, such as tamoxifen, in the aquatic environment. © 2015 Wiley Periodicals, Inc. Environ Toxicol, 2015. PMID: 26303313 [PubMed - as supplied by publisher]

Long-term effect of early postnatal overnutrition on insulin resistance and serum fatty acid profiles in male rats. Lipids Health Dis. 2015;14(1):96 Authors: Bei F, Jia J, Jia YQ, Sun JH, Liang F, Yu ZY, Cai W Abstract BACKGROUND: Increasing evidence suggests that overnutrition during the early postnatal period, a critical window of development, increases the risk of adult-onset obesity and insulin resistance. In this study, we investigated the impact of overnutrition during the suckling period on body weight, serum biochemistry and serum fatty acid metabolomics in male rats. METHODS: Rats raised in small litters (SL, 3 pups/dam) and normal litters (NL, 10 pups/dam) were used to model early postnatal overnutrition and control, respectively. Serum glucose, triglyceride, high-density lipoprotein-cholesterol, free fatty acid, insulin and leptin concentrations were assayed using standard biochemical techniques. Serum fatty acids were identified and quantified using a gas chromatography-mass spectrometry-based metabolomic approach. mRNA and protein levels of key components of the insulin receptor signaling pathway were measured in epididymal fat and gastrocnemius muscle by quantitative PCR and western blotting. RESULTS: SL rats were 37.3 % and 15.1 % heavier than NL rats at weaning and 16-weeks-old, respectively. They had increased visceral fat mass, adult-onset insulin resistance and glucose intolerance as well as elevated serum levels of free fatty acids and triglycerides. All detectable fatty acids were elevated in the serum of SL pups at weaning compared to NL controls, and significant increases in the levels of four fatty acids (palmitic acid, palmitoleic acid, oleic acid and arachidonic acid) persisted into adulthood. Moreover, a significantly positive correlation was identified between an insulin resistance index (HOMA-IR) and concentrations of myristic, palmitic, palmitoleic and oleic acid in serum at postnatal 16 weeks. Early postnatal overnutrition also resulted in a significant downregulation of insulin receptor substrate-1 (Irs-1), protein kinase B (Akt2) and glucose transporter 4 (Glut4) at the protein level in epididymal fat of SL rats at 16 weeks, accompanied by decreased mRNA levels for Irs-1 and Glut4. In gastrocnemius muscle, Akt2 and Glut4 mRNA and Glut4 protein levels were significantly decreased in SL rats. CONCLUSIONS: This study demonstrates that early postnatal overnutrition can have long-lasting effects on body weight and serum fatty acid profiles and can lead to impaired insulin signaling pathway in visceral white adipose tissue and skeletal muscle, which may play a major role in IR. PMID: 26302954 [PubMed - in process]

Related Articles CANCER. A p53-regulated immune checkpoint relevant to cancer. Science. 2015 Jul 31;349(6247):476-7 Authors: Zitvogel L, Kroemer G PMID: 26228128 [PubMed - indexed for MEDLINE]

Related Articles n-3 fatty acids effectively improve the reference memory-related learning ability associated with increased brain docosahexaenoic acid-derived docosanoids in aged rats. Biochim Biophys Acta. 2015 Feb;1851(2):203-9 Authors: Hashimoto M, Katakura M, Tanabe Y, Al Mamun A, Inoue T, Hossain S, Arita M, Shido O Abstract We investigated whether a highly purified eicosapentaenoic acid (EPA) and a concentrated n-3 fatty acid formulation (prescription TAK-085) containing EPA and docosahexaenoic acid (DHA) ethyl ester could improve the learning ability of aged rats and whether this specific outcome had any relation with the brain levels of EPA-derived eicosanoids and DHA-derived docosanoids. The rats were tested for reference memory errors (RMEs) and working memory errors (WMEs) in an eight-arm radial maze. Fatty acid compositions were analyzed by GC, whereas brain eicosanoid/docosanoids were measured by LC-ESI-MS-MS-based analysis. The levels of lipid peroxides (LPOs) were measured by thiobarbituric acid reactive substances. The administration of TAK-085 at 300 mg·kg⁻¹day⁻¹ for 17 weeks reduced the number of RMEs in aged rats compared with that in the control rats. Both TAK-085 and EPA administration increased plasma EPA and DHA levels in aged rats, with concurrent increases in DHA and decreases in arachidonic acid in the corticohippocampal brain tissues. TAK-085 administration significantly increased the formation of EPA-derived 5-HETE and DHA-derived 7-, 10-, and 17-HDoHE, PD1, RvD1, and RvD2. ARA-derived PGE2, PGD2, and PGF2α significantly decreased in TAK-085-treated rats. DHA-derived mediators demonstrated a significantly negative correlation with the number of RMEs, whereas EPA-derived mediators did not exhibit any relationship. Furthermore, compared with the control rats, the levels of LPO in the plasma, cerebral cortex, and hippocampus were significantly reduced in TAK-085-treated rats. The findings of the present study suggest that long-term EPA+DHA administration may be a possible preventative strategy against age-related cognitive decline. PMID: 25450447 [PubMed - indexed for MEDLINE]

Related Articles Simultaneous UHPLC/DAD/(+/-)HESI-MS/MS analysis of phenolic acids and nepetalactones in methanol extracts of Nepeta species: a possible application in chemotaxonomic studies. Phytochem Anal. 2015 Jan-Feb;26(1):72-85 Authors: Mišić D, Siler B, Gašić U, Avramov S, Zivković S, Nestorović Živković J, Milutinović M, Tešić Z Abstract INTRODUCTION: Nepeta species contain a variety of secondary metabolites, including iridoid monoterpenes - nepetalactones and phenolic acids - that are considered the main bioactive constituents. This work represents the first attempt to comparatively explore variations in these two major groups of secondary metabolites within the genus. OBJECTIVE: To develop an efficient analytical methodology for simultaneous analysis of nepetalactones and phenolic acids in methanol extracts of selected Nepeta species, and to evaluate its potential application in chemotaxonomic studies. MATERIAL AND METHODS: A UHPLC combined with linear-trap quadrupole (LTQ) orbitrap MS method was used to characterise chemical diversity and complexity of phenolics among 12 selected Nepeta species. A targeted metabolomic approach using UHPLC coupled to a diode array detector (DAD) and combined with (+/-) heated electrospray ionisation (HESI) MS/MS was developed and validated for quantitative analysis of six hydroxycinnamic acid derivatives and four nepetalactones. RESULTS: Phenolic profiling provided a valuable database of bioactive compounds in the plant group studied, including phenolic acids (hydroxybenzoic and hydroxycinnamic acids) and flavonoids (flavones, flavonols and flavanones). Principal component analysis and cluster analysis suggested the applicability of 10 targeted compounds as chemomarkers for chemotaxonomic studies. Pearson's correlation analysis revealed significant positive correlations between metabolites involved in different biosynthetic pathways (phenylpropanoid or monoterpenoid). CONCLUSION: The described targeted metabolomic approach proved to be highly beneficial in designing a phytochemical overview of the genus Nepeta, and might have applications in further clarification of phylogenetic relations. Furthermore, it has the potential to be implemented in a routine quality control of plant material and herbal preparations. PMID: 25431035 [PubMed - indexed for MEDLINE]

Related Articles Revealing metabolomic variations in Cortex Moutan from different root parts using HPLC-MS method. Phytochem Anal. 2015 Jan-Feb;26(1):86-93 Authors: Xiao C, Wu M, Chen Y, Zhang Y, Zhao X, Zheng X Abstract INTRODUCTION: The distribution of metabolites in the different root parts of Cortex Moutan (the root bark of Paeonia suffruticosa Andrews) is not well understood, therefore, scientific evidence is not available for quality assessment of Cortex Moutan. OBJECTIVE: To reveal metabolomic variations in Cortex Moutan in order to gain deeper insights to enable quality control. METHODS: Metabolomic variations in the different root parts of Cortex Moutan were characterised using high-performance liquid chromatography combined with mass spectrometry (HPLC-MS) and multivariate data analysis. The discriminating metabolites in different root parts were evaluated by the one-way analysis of variance and a fold change parameter. RESULTS: The metabolite profiles of Cortex Moutan were largely dominated by five primary and 41 secondary metabolites . Higher levels of malic acid, gallic acid and mudanoside-B were mainly observed in the second lateral roots, whereas dihydroxyacetophenone, benzoyloxypaeoniflorin, suffruticoside-A, kaempferol dihexoside, mudanpioside E and mudanpioside J accumulated in the first lateral and axial roots. The highest contents of paeonol, galloyloxypaeoniflorin and procyanidin B were detected in the axial roots. Accordingly, metabolite compositions of Cortex Moutan were found to vary among different root parts. CONCLUSION: The axial roots have higher quality than the lateral roots in Cortex Moutan due to the accumulation of bioactive secondary metabolites associated with plant physiology. These findings provided important scientific evidence for grading Cortex Moutan on the general market. Copyright © 2014 John Wiley & Sons, Ltd. PMID: 25230378 [PubMed - indexed for MEDLINE]

Related Articles Qualitative and spatial metabolite profiling of lichens by a LC-MS approach combined with optimised extraction. Phytochem Anal. 2015 Jan-Feb;26(1):23-33 Authors: Parrot D, Peresse T, Hitti E, Carrie D, Grube M, Tomasi S Abstract INTRODUCTION: Lichens are self-sustaining partnerships comprising fungi as shape-forming partners for their enclosed symbiotic algae. They produce a tremendous diversity of metabolites (1050 metabolites described so far). OBJECTIVES: A comparison of metabolic profiles in nine lichen species belonging to three genera (Lichina, Collema and Roccella) by using an optimised extraction protocol, determination of the fragmentation pathway and the in situ localisation for major compounds in Roccella species. METHODS: Chemical analysis was performed using a complementary study combining a Taguchi experimental design with qualitative analysis by high-performance liquid chromatography coupled with mass spectrometry techniques. RESULTS: Optimal conditions to obtain the best total extraction yield were determined as follows: mortar grinding to a fine powder, two successive extractions, solid:liquid ratio (2:60) and 700 rpm stirring. Qualitative analysis of the metabolite profiling of these nine species extracted with the optimised method was corroborated using MS and MS/MS approaches. Nine main compounds were identified: 1 β-orcinol, 2 orsellinic acid, 3 putative choline sulphate, 4 roccellic acid, 5 montagnetol, 6 lecanoric acid, 7 erythrin, 8 lepraric acid and 9 acetylportentol, and several other compounds were reported. Identification was performed using the m/z ratio, fragmentation pathway and/or after isolation by NMR analysis. The variation of the metabolite profile in differently organised parts of two Roccella species suggests a specific role of major compounds in developmental stages of this symbiotic association. CONCLUSION: Metabolic profiles represent specific chemical species and depend on the extraction conditions, the kind of the photobiont partner and the in situ localisation of major compounds. PMID: 25130294 [PubMed - indexed for MEDLINE]

Related Articles Eosinophils control the resolution of inflammation and draining lymph node hypertrophy through the proresolving mediators and CXCL13 pathway in mice. FASEB J. 2014 Sep;28(9):4036-43 Authors: Tani Y, Isobe Y, Imoto Y, Segi-Nishida E, Sugimoto Y, Arai H, Arita M Abstract Resolution of inflammation is critical to restoration of tissue function after an inflammatory response. We previously demonstrated that 12/15-lipoxygenase (12/15-LOX)-expressing eosinophils contribute to this process in murine zymosan-induced peritonitis. In this study, eosinophils promoted resolution by regulating expression of macrophage CXCL13. Microarray analysis revealed that eosinophils significantly increased (∼3-fold) the expression of macrophage CXCL13 by a 12/15-LOX-dependent mechanism. CXCL13 depletion caused a resolution defect, with the reduced appearance of phagocytes carrying engulfed zymosan in the draining lymph nodes. Inflamed lymph node hypertrophy, a critical feature of the resolution process, was reduced by ∼60% in eosinophil-deficient mice, and adoptive transfer of eosinophils or administration of CXCL13 corrected this defect. Administration of the 12/15-LOX-derived mediator lipoxin A4 (LXA4) increased the expression of CXCL13 and restored the defect of lymph node hypertrophy in eosinophil-deficient mice. These results demonstrate that eosinophils control the resolution of inflammation and draining lymph node hypertrophy through proresolving lipid mediators and the CXCL13 pathway in mice. PMID: 24891522 [PubMed - indexed for MEDLINE]

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Ambient ionization MS for bioanalysis: recent developments and challenges. Bioanalysis. 2015 Aug;7(15):1901-1923 Authors: Takyi-Williams J, Liu CF, Tang K Abstract Ambient ionization MS has become very popular in analytical science and has now evolved as an effective analytical tool in metabolomics, biological tissue imaging, protein and small molecule drug analysis, where biological samples are probed in a rapid and direct fashion with minimal sample preparation at ambient conditions. However, certain inherent challenges continue to hinder the vibrant prospects of these methods for in situ analyses or to replace conventional methods in bioanalysis. This review provides an introduction to the field and its application in bioanalysis, with an emphasis on the most recent developments and applications. Furthermore, ongoing challenges or limitations related to quantitation, sensitivity, selectivity, instrumentation and mass range of these ambient methods will also be discussed. PMID: 26295990 [PubMed - as supplied by publisher]

Effects of Graphene Oxide and Oxidized Carbon Nanotubes on the Cellular Division, Microstructure, Uptake, Oxidative Stress and Metabolic Profiles. Environ Sci Technol. 2015 Aug 21; Authors: Hu X, Ouyang S, Mu L, An J, Zhou Q Abstract Nanomaterial oxides are common formations of nanomaterials in the natural environment. Herein, the nanotoxicology of typical graphene oxide (GO) and carboxyl single-walled carbon nanotubes (C-SWCNT) was compared. The results showed that cell division of Chlorella vulgaris was promoted at 24 h and then inhibited at 96 h after nanomaterial exposure. At 96 h, GO and C-SWCNT inhibited the rates of cell division by 0.08-15% and 0.8-28.3%, respectively. Both GO and C-SWCNT covered the cell surface, but the uptake percentage of C-SWCNT was 2-fold higher than that of GO. C-SWCNT induced stronger plasmolysis and mitochondrial membrane potential loss and decreased the cell viability to a greater extent than GO. Moreover, C-SWCNT-exposed cells exhibited more starch grains and lysosome formation and higher reactive oxygen species (ROS) levels than GO-exposed cells. Metabolomics analysis revealed significant differences in the metabolic profiles among the control, C-SWCNT and GO groups. The metabolisms of alkanes, lysine, octadecadienoic acid and valine was associated with ROS and could be considered as new biomarkers of ROS. The nanotoxicological mechanisms involved the inhibition of fatty acid, amino acid and small molecule acid metabolisms. These findings provide new insights into the effects of GO and C-SWCNT on cellular responses. PMID: 26295980 [PubMed - as supplied by publisher]