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Sample records for acid metabolites contributes

  1. Inhibition of Human Hepatic Bile Acid Transporters by Tolvaptan and Metabolites: Contributing Factors to Drug-Induced Liver Injury?

    Slizgi, Jason R; Lu, Yang; Brouwer, Kenneth R; St Claire, Robert L; Freeman, Kimberly M; Pan, Maxwell; Brock, William J; Brouwer, Kim L R

    2016-01-01

    Tolvaptan is a vasopressin V(2)-receptor antagonist that has shown promise in treating Autosomal Dominant Polycystic Kidney Disease (ADPKD). Tolvaptan was, however, associated with liver injury in some ADPKD patients. Inhibition of bile acid transporters may be contributing factors to drug-induced liver injury. In this study, the ability of tolvaptan and two metabolites, DM-4103 and DM-4107, to inhibit human hepatic transporters (NTCP, BSEP, MRP2, MRP3, and MRP4) and bile acid transport in sandwich-cultured human hepatocytes (SCHH) was explored. IC(50) values were determined for tolvaptan, DM-4103 and DM-4107 inhibition of NTCP (∼41.5, 16.3, and 95.6 μM, respectively), BSEP (31.6, 4.15, and 119 μM, respectively), MRP2 (>50, ∼51.0, and >200 μM, respectively), MRP3 (>50, ∼44.6, and 61.2 μM, respectively), and MRP4 (>50, 4.26, and 37.9 μM, respectively). At the therapeutic dose of tolvaptan (90 mg), DM-4103 exhibited a C(max)/IC(50) value >0.1 for NTCP, BSEP, MRP2, MRP3, and MRP4. Tolvaptan accumulation in SCHH was extensive and not sodium-dependent; intracellular concentrations were ∼500 μM after a 10-min incubation duration with tolvaptan (15 μM). The biliary clearance of taurocholic acid (TCA) decreased by 43% when SCHH were co-incubated with tolvaptan (15 μM) and TCA (2.5 μM). When tolvaptan (15 μM) was co-incubated with 2.5 μM of chenodeoxycholic acid, taurochenodeoxycholic acid, or glycochenodeoxycholic acid in separate studies, the cellular accumulation of these bile acids increased by 1.30-, 1.68-, and 2.16-fold, respectively. Based on these data, inhibition of hepatic bile acid transport may be one of the biological mechanisms underlying tolvaptan-associated liver injury in patients with ADPKD. PMID:26507107

  2. Arachidonic acid metabolites in pathogenic yeasts

    Ells Ruan

    2012-08-01

    Full Text Available Abstract Although most of what is known about the biology and function of arachidonic acid metabolites comes from the study of mammalian biology, these compounds can also be produced by lower eukaryotes, including yeasts and other fungi. It is also in this group of organisms that the least is known about the metabolic pathways leading to the production of these compounds as well as the functions of these compounds in the biology of fungi and yeasts. This review will deal with the discovery of oxylipins from polyunsaturated fatty acids, and more specifically the arachidonic acid derived eicosanoids, such as 3-hydroxy eicosatetraenoic acid, prostaglandin F2α and prostaglandin E2, in yeasts starting in the early 1990s. This review will also focus on what is known about the metabolic pathways and/or proteins involved in the production of these compounds in pathogenic yeasts. The possible roles of these compounds in the biology, including the pathology, of these organisms will be discussed.

  3. Biodegradation of clofibric acid and identification of its metabolites

    Salgado, R. [REQUIMTE/CQFB, Chemistry Department, FCT, Universidade Nova de Lisboa, 2829-516 Caparica (Portugal); ESTS-IPS, Escola Superior de Tecnologia de Setubal do Instituto Politecnico de Setubal, Rua Vale de Chaves, Campus do IPS, Estefanilha, 2910-761 Setubal (Portugal); Oehmen, A. [REQUIMTE/CQFB, Chemistry Department, FCT, Universidade Nova de Lisboa, 2829-516 Caparica (Portugal); Carvalho, G. [REQUIMTE/CQFB, Chemistry Department, FCT, Universidade Nova de Lisboa, 2829-516 Caparica (Portugal); Instituto de Biologia Experimental e Tecnologica (IBET), Av. da Republica (EAN), 2784-505 Oeiras (Portugal); Noronha, J.P. [REQUIMTE/CQFB, Chemistry Department, FCT, Universidade Nova de Lisboa, 2829-516 Caparica (Portugal); Reis, M.A.M., E-mail: amr@fct.unl.pt [REQUIMTE/CQFB, Chemistry Department, FCT, Universidade Nova de Lisboa, 2829-516 Caparica (Portugal)

    2012-11-30

    Graphical abstract: Metabolites produced during clofibric acid biodegradation. Highlights: Black-Right-Pointing-Pointer Clofibric acid is biodegradable. Black-Right-Pointing-Pointer Mainly heterotrophic bacteria degraded the clofibric acid. Black-Right-Pointing-Pointer Metabolites of clofibric acid biodegradation were identified. Black-Right-Pointing-Pointer The metabolic pathway of clofibric acid biodegradation is proposed. - Abstract: Clofibric acid (CLF) is the pharmaceutically active metabolite of lipid regulators clofibrate, etofibrate and etofyllinclofibrate, and it is considered both environmentally persistent and refractory. This work studied the biotransformation of CLF in aerobic sequencing batch reactors (SBRs) with mixed microbial cultures, monitoring the efficiency of biotransformation of CLF and the production of metabolites. The maximum removal achieved was 51% biodegradation (initial CLF concentration = 2 mg L{sup -1}), where adsorption and abiotic removal mechanisms were shown to be negligible, showing that CLF is indeed biodegradable. Tests showed that the observed CLF biodegradation was mainly carried out by heterotrophic bacteria. Three main metabolites were identified, including {alpha}-hydroxyisobutyric acid, lactic acid and 4-chlorophenol. The latter is known to exhibit higher toxicity than the parent compound, but it did not accumulate in the SBRs. {alpha}-Hydroxyisobutyric acid and lactic acid accumulated for a period, where nitrite accumulation may have been responsible for inhibiting their degradation. A metabolic pathway for the biodegradation of CLF is proposed in this study.

  4. Effects of fluticasone propionate inhalation on levels of arachidonic acid metabolites in patients with chronic obstructive pulmonary disease

    Gert T. Verhoeven

    2001-01-01

    Full Text Available Background: In smoking COPD patients the bronchoalveolar lavage (BAL fluid contains high numbers of inflammatory cells. These cells might produce arachidonic acid (AA metabolites, which contribute to inflammation and an increased bronchomotor tone.

  5. Seed coat color and seed weight contribute differential responses of targeted metabolites in soybean seeds.

    Lee, Jinwook; Hwang, Young-Sun; Kim, Sun Tae; Yoon, Won-Byong; Han, Won Young; Kang, In-Kyu; Choung, Myoung-Gun

    2017-01-01

    The distribution and variation of targeted metabolites in soybean seeds are affected by genetic and environmental factors. In this study, we used 192 soybean germplasm accessions collected from two provinces of Korea to elucidate the effects of seed coat color and seeds dry weight on the metabolic variation and responses of targeted metabolites. The effects of seed coat color and seeds dry weight were present in sucrose, total oligosaccharides, total carbohydrates and all measured fatty acids. The targeted metabolites were clustered within three groups. These metabolites were not only differently related to seeds dry weight, but also responded differentially to seed coat color. The inter-relationship between the targeted metabolites was highly present in the result of correlation analysis. Overall, results revealed that the targeted metabolites were diverged in relation to seed coat color and seeds dry weight within locally collected soybean seed germplasm accessions. PMID:27507473

  6. Gut Microbial Fatty Acid Metabolites Reduce Triacylglycerol Levels in Hepatocytes.

    Nanthirudjanar, Tharnath; Furumoto, Hidehiro; Zheng, Jiawen; Kim, Young-Il; Goto, Tsuyoshi; Takahashi, Nobuyuki; Kawada, Teruo; Park, Si-Bum; Hirata, Akiko; Kitamura, Nahoko; Kishino, Shigenobu; Ogawa, Jun; Hirata, Takashi; Sugawara, Tatsuya

    2015-11-01

    Hydroxy and oxo fatty acids were recently found to be produced as intermediates during gut microbial fatty acid metabolism. Lactobacillus plantarum produces these fatty acids from unsaturated fatty acids such as linoleic acid. In this study, we investigated the effects of these gut microbial fatty acid metabolites on the lipogenesis in liver cells. We screened their effect on sterol regulatory element binding protein-1c (SREBP-1c) expression in HepG2 cells treated with a synthetic liver X receptor α (LXRα) agonist (T0901317). The results showed that 10-hydroxy-12(Z)-octadecenoic acid (18:1) (HYA), 10-hydroxy-6(Z),12(Z)-octadecadienoic acid (18:2) (γHYA), 10-oxo-12(Z)-18:1 (KetoA), and 10-oxo-6(Z),12(Z)-18:2 (γKetoA) significantly decreased SREBP-1c mRNA expression induced by T0901317. These fatty acids also downregulated the mRNA expression of lipogenic genes by suppressing LXRα activity and inhibiting SREBP-1 maturation. Oral administration of KetoA, which effectively reduced triacylglycerol accumulation and acetyl-CoA carboxylase 2 (ACC2) expression in HepG2 cells, for 2 weeks significantly decreased Srebp-1c, Scd-1, and Acc2 expression in the liver of mice fed a high-sucrose diet. Our findings suggest that the hypolipidemic effect of the fatty acid metabolites produced by L. plantarum can be exploited in the treatment of cardiovascular diseases or dyslipidemia. PMID:26399511

  7. Structural Analysis of Metabolites of Asiatic Acid and Its Analogue Madecassic Acid in Zebrafish Using LC/IT-MSn

    Binbin Xia

    2015-02-01

    Full Text Available Although zebrafish has become a significant animal model for drug discovery and screening, drug metabolism in zebrafish remains largely unknown. Asiatic acid (AA and madecassic acid (MA, two natural pentacyclic triterpenoids mainly obtained from Centella asiatica (L. Urban, have been found to possess many pharmacological effects. This study is to probe the metabolic capability of zebrafish via investigation of the drug metabolism of AA and MA in zebrafish, using a sensitive LC/IT-MSn method. In addition, the main fragmentation pathways of AA and MA were reported for the first time. Nineteen metabolites of AA and MA were firstly identified after zebrafish was exposed to the drug, which all were the phase I metabolites and mainly formed from hydroxylation, dehydrogenation, hydroxylation and dehydrogenation, dihydroxylation and dehydrogenation, and dehydroxylation reaction. The results indicated that zebrafish possessed strong metabolic capacity, and the metabolites of AA and MA were formed via similar metabolic pathways and well matched with the known metabolic rules in vivo and in vitro, which supports the widely use of this system in drug metabolism research. This investigation would also contribute to the novel information on the structural elucidation, in vivo metabolites and metabolic mechanism of pentacyclic triterpenoids.

  8. Contribution of Network Connectivity in Determining the Relationship between Gene Expression and Metabolite Concentration Changes

    Zelezniak, Aleksej; Sheridan, Steven; Patil, Kiran Raosaheb

    2014-01-01

    of reaction kinetics in metabolite concentration control is well studied at the level of individual reactions, the contribution of network connectivity has remained relatively unclear. Here we report a modeling framework that integrates both reaction kinetics and network connectivity constraints...... for describing the interplay between metabolite concentrations and mRNA levels. We used this framework to investigate correlations between the gene expression and the metabolite concentration changes in Saccharomyces cerevisiae during its metabolic cycle, as well as in response to three fundamentally different......One of the primary mechanisms through which a cell exerts control over its metabolic state is by modulating expression levels of its enzyme-coding genes. However, the changes at the level of enzyme expression allow only indirect control over metabolite levels, for two main reasons. First...

  9. Mixture toxicity of the antiviral drug Tamiflu (oseltamivir ethylester) and its active metabolite oseltamivir acid

    Escher, Beate I., E-mail: b.escher@uq.edu.au [University of Queensland, National Research Centre for Environmental Toxicology (Entox), 39 Kessels Rd, Brisbane, Qld 4108 (Australia); Eawag, Swiss Federal Institute of Aquatic Science and Technology, 8600 Duebendorf (Switzerland); Bramaz, Nadine; Lienert, Judit; Neuwoehner, Judith [Eawag, Swiss Federal Institute of Aquatic Science and Technology, 8600 Duebendorf (Switzerland); Straub, Juerg Oliver [F.Hoffmann-La Roche Ltd, Corporate Safety, Health and Environmental Protection, 4070 Basel (Switzerland)

    2010-02-18

    Tamiflu (oseltamivir ethylester) is an antiviral agent for the treatment of influenza A and B. The pro-drug Tamiflu is converted in the human body to the pharmacologically active metabolite, oseltamivir acid, with a yield of 75%. Oseltamivir acid is indirectly photodegradable and slowly biodegradable in sewage works and sediment/water systems. A previous environmental risk assessment has concluded that there is no bioaccumulation potential of either of the compounds. However, little was known about the ecotoxicity of the metabolite. Ester hydrolysis typically reduces the hydrophobicity and thus the toxicity of a compound. In this case, a zwitterionic, but overall neutral species is formed from the charged parent compound. If the speciation and predicted partitioning into biological membranes is considered, the metabolite may have a relevant contribution to the overall toxicity. These theoretical considerations triggered a study to investigate the toxicity of oseltamivir acid (OA), alone and in binary mixtures with its parent compound oseltamivir ethylester (OE). OE and OA were found to be baseline toxicants in the bioluminescence inhibition test with Vibrio fischeri. Their mixture effect lay between predictions for concentration addition and independent action for the mixture ratio excreted in urine and nine additional mixture ratios of OE and OA. In contrast, OE was an order of magnitude more toxic than OA towards algae, with a more pronounced effect when the direct inhibition of photosystem II was used as toxicity endpoint opposed to the 24 h growth rate endpoint. The binary mixtures in this assay yielded experimental mixture effects that agreed with predictions for independent action. This is consistent with the finding that OE exhibits slightly enhanced toxicity, while OA acts as baseline toxicant. Therefore, with respect to mixture classification, the two compounds can be considered as acting according to different modes of toxic action, although there are

  10. Mixture toxicity of the antiviral drug Tamiflu (oseltamivir ethylester) and its active metabolite oseltamivir acid

    Tamiflu (oseltamivir ethylester) is an antiviral agent for the treatment of influenza A and B. The pro-drug Tamiflu is converted in the human body to the pharmacologically active metabolite, oseltamivir acid, with a yield of 75%. Oseltamivir acid is indirectly photodegradable and slowly biodegradable in sewage works and sediment/water systems. A previous environmental risk assessment has concluded that there is no bioaccumulation potential of either of the compounds. However, little was known about the ecotoxicity of the metabolite. Ester hydrolysis typically reduces the hydrophobicity and thus the toxicity of a compound. In this case, a zwitterionic, but overall neutral species is formed from the charged parent compound. If the speciation and predicted partitioning into biological membranes is considered, the metabolite may have a relevant contribution to the overall toxicity. These theoretical considerations triggered a study to investigate the toxicity of oseltamivir acid (OA), alone and in binary mixtures with its parent compound oseltamivir ethylester (OE). OE and OA were found to be baseline toxicants in the bioluminescence inhibition test with Vibrio fischeri. Their mixture effect lay between predictions for concentration addition and independent action for the mixture ratio excreted in urine and nine additional mixture ratios of OE and OA. In contrast, OE was an order of magnitude more toxic than OA towards algae, with a more pronounced effect when the direct inhibition of photosystem II was used as toxicity endpoint opposed to the 24 h growth rate endpoint. The binary mixtures in this assay yielded experimental mixture effects that agreed with predictions for independent action. This is consistent with the finding that OE exhibits slightly enhanced toxicity, while OA acts as baseline toxicant. Therefore, with respect to mixture classification, the two compounds can be considered as acting according to different modes of toxic action, although there are

  11. From Dietary Fiber to Host Physiology: Short-Chain Fatty Acids as Key Bacterial Metabolites.

    Koh, Ara; De Vadder, Filipe; Kovatcheva-Datchary, Petia; Bäckhed, Fredrik

    2016-06-01

    A compelling set of links between the composition of the gut microbiota, the host diet, and host physiology has emerged. Do these links reflect cause-and-effect relationships, and what might be their mechanistic basis? A growing body of work implicates microbially produced metabolites as crucial executors of diet-based microbial influence on the host. Here, we will review data supporting the diverse functional roles carried out by a major class of bacterial metabolites, the short-chain fatty acids (SCFAs). SCFAs can directly activate G-coupled-receptors, inhibit histone deacetylases, and serve as energy substrates. They thus affect various physiological processes and may contribute to health and disease. PMID:27259147

  12. Purification and H-1 NMR spectroscopic characterization of phase II metabolites of tolfenamic acid

    Sidelmann, U. G.; Christiansen, E.; Krogh, L.;

    1997-01-01

    acid; the study shows the applicability of H-1 NMR for the identification of drug metabolites in biological fluids. In addition to NMR analysis, two metabolites were also identified by mass spectrometry (MS), The glucuronides of the following parent compounds, N-(2-methyl-3-chlorophenyl...... endogenous polar compounds that are present in the urine. The individual metabolites were purified by preparative high performance liquid chromatography (HPLC) and then identified using H-1 NMR, Both one- and two-dimensional NMR experiments were performed to identify the phase II metabolites of tolfenamic......), and N-(2-methyl-4-hydroxyphenyl)-anthranilic acid (11) were identified. The phase II metabolites (5-11) had not previously been identified in urine from humans administered tolfenamic acid. The phase I metabolites of the glucuronides 7, 8, 10, and 11 were identified here for the first time. An HPLC...

  13. Biological Significance of Urolithins, the Gut Microbial Ellagic Acid-Derived Metabolites: The Evidence So Far

    Espín, Juan Carlos; Larrosa, Mar; García-Conesa, María Teresa; Tomás-Barberán, Francisco

    2013-01-01

    The health benefits attributed to pomegranate have been associated with its high content in polyphenols, particularly ellagitannins. This is also the case for other ellagitannin-containing fruits and nuts including strawberry, raspberry, blackberry, walnuts, and muscadine grapes. The bioavailability of ellagitannins and ellagic acid is however very low. These molecules suffer extensive metabolism by the gut microbiota to produce urolithins that are much better absorbed. Urolithins circulate in plasma as glucuronide and sulfate conjugates at concentrations in the range of 0.2–20 μM. It is therefore conceivable that the health effects of ellagitannin-containing products can be associated with these gut-produced urolithins, and thus the evaluation of the biological effects of these metabolites is essential. Recent research, mostly based on in vitro testing, has shown preliminary evidence of the anti-inflammatory, anticarcinogenic, antiglycative, antioxidant, and antimicrobial effects of urolithins, supporting their potential contribution to the health effects attributed to pomegranate and ellagitannin-rich foods. The number of in vivo studies is still limited, but they show preventive effects of urolithins on gut and systemic inflammation that encourage further research. Both in vivo and mechanistic studies are necessary to clarify the health effects of these metabolites. Attention should be paid when designing these mechanistic studies in order to use the physiologically relevant metabolites (urolithins in gut models and their conjugated derivatives in systemic models) at concentrations that can be reached in vivo. PMID:23781257

  14. Bacterial dynamics and metabolite changes in solid-state acetic acid fermentation of Shanxi aged vinegar.

    Li, Sha; Li, Pan; Liu, Xiong; Luo, Lixin; Lin, Weifeng

    2016-05-01

    Solid-state acetic acid fermentation (AAF), a natural or semi-controlled fermentation process driven by reproducible microbial communities, is an important technique to produce traditional Chinese cereal vinegars. Highly complex microbial communities and metabolites are involved in traditional Chinese solid-state AAF, but the association between microbiota and metabolites during this process are still poorly understood. In this study, we performed amplicon 16S rRNA gene sequencing on the Illumina MiSeq platform, PCR-denaturing gradient gel electrophoresis, and metabolite analysis to trace the bacterial dynamics and metabolite changes under AAF process. A succession of bacterial assemblages was observed during the AAF process. Lactobacillales dominated all the stages. However, Acetobacter species in Rhodospirillales were considerably accelerated during AAF until the end of fermentation. Quantitative PCR results indicated that the biomass of total bacteria showed a "system microbe self-domestication" process in the first 3 days, and then peaked at the seventh day before gradually decreasing until the end of AAF. Moreover, a total of 88 metabolites, including 8 organic acids, 16 free amino acids, and 66 aroma compounds were detected during AAF. Principal component analysis and cluster analyses revealed the high correlation between the dynamics of bacterial community and metabolites. PMID:26754813

  15. Separation and detection of amino acid metabolites of Escherichia coli in microbial fuel cell with CE.

    Wang, Wei; Ma, Lihong; Lin, Ping; Xu, Kaixuan

    2016-07-01

    In this work, CE-LIF was employed to investigate the amino acid metabolites produced by Escherichia coli (E. coli) in microbial fuel cell (MFC). Two peptides, l-carnosine and l-alanyl-glycine, together with six amino acids, cystine, alanine, lysine, methionine, tyrosine, arginine were separated and detected in advance by a CE-LIF system coupled with a homemade spontaneous injection device. The injection device was devised to alleviate the effect of electrical discrimination for analytes during sample injection. All analytes could be completely separated within 8 min with detection limits of 20-300 nmol/L. Then this method was applied to analyze the substrate solution containing amino acid metabolites produced by E. coli. l-carnosine, l-alanyl-glycine, and cystine were used as the carbon, nitrogen, and sulfur source for the E. coli culture in the MFC to investigate the amino acid metabolites during metabolism. Two MFCs were used to compare the activity of metabolism of the bacteria. In the sample collected at the running time 200 h of MFC, the amino acid methionine was discovered as the metabolite with the concentrations 23.3 μg/L. PMID:27121957

  16. Heat generates oxidized linoleic acid metabolites that activate TRPV1 and produce pain in rodents

    Patwardhan, Amol M.; Akopian, Armen N.; Ruparel, Nikita B.; Diogenes, Anibal; Weintraub, Susan T; Uhlson, Charis; Murphy, Robert C.; Hargreaves, Kenneth M.

    2010-01-01

    The transient receptor potential vanilloid 1 (TRPV1) channel is the principal detector of noxious heat in the peripheral nervous system. TRPV1 is expressed in many nociceptors and is involved in heat-induced hyperalgesia and thermoregulation. The precise mechanism or mechanisms mediating the thermal sensitivity of TRPV1 are unknown. Here, we have shown that the oxidized linoleic acid metabolites 9- and 13-hydroxyoctadecadienoic acid (9- and 13-HODE) are formed in mouse and rat skin biopsies b...

  17. Microbial diversity and metabolite composition of Belgian red-brown acidic ales.

    Snauwaert, Isabel; Roels, Sanne P; Van Nieuwerburg, Filip; Van Landschoot, Anita; De Vuyst, Luc; Vandamme, Peter

    2016-03-16

    Belgian red-brown acidic ales are sour and alcoholic fermented beers, which are produced by mixed-culture fermentation and blending. The brews are aged in oak barrels for about two years, after which mature beer is blended with young, non-aged beer to obtain the end-products. The present study evaluated the microbial community diversity of Belgian red-brown acidic ales at the end of the maturation phase of three subsequent brews of three different breweries. The microbial diversity was compared with the metabolite composition of the brews at the end of the maturation phase. Therefore, mature brew samples were subjected to 454 pyrosequencing of the 16S rRNA gene (bacteria) and the internal transcribed spacer region (yeasts) and a broad range of metabolites was quantified. The most important microbial species present in the Belgian red-brown acidic ales investigated were Pediococcus damnosus, Dekkera bruxellensis, and Acetobacter pasteurianus. In addition, this culture-independent analysis revealed operational taxonomic units that were assigned to an unclassified fungal community member, Candida, and Lactobacillus. The main metabolites present in the brew samples were L-lactic acid, D-lactic acid, and ethanol, whereas acetic acid was produced in lower quantities. The most prevailing aroma compounds were ethyl acetate, isoamyl acetate, ethyl hexanoate, and ethyl octanoate, which might be of impact on the aroma of the end-products. PMID:26802571

  18. Metabolite Profiling of Sugarcane Genotypes and Identification of Flavonoid Glycosides and Phenolic Acids.

    Coutinho, Isabel D; Baker, John M; Ward, Jane L; Beale, Michael H; Creste, Silvana; Cavalheiro, Alberto J

    2016-06-01

    Sugarcane is an important agricultural crop in the economy of tropical regions, and Brazil has the largest cultivated acreage in the world. Sugarcane accumulates high levels of sucrose in its stalks. Other compounds produced by sugarcane are currently not of economic importance. To explore potential coproducts, we have studied the chemical diversity of sugarcane genotypes, via metabolite profiling of leaves by NMR and LC-DAD-MS. Metabolites were identified via in-house and public databases. From the analysis of 60 HPLC-fractionated extracts, LC-DAD-MS detected 144 metabolites, of which 56 were identified (MS-MS and (1)H NMR), including 19 phenolics and 25 flavones, with a predominance of isomeric flavone C-glycosides. Multivariate analysis of the profiles from genotypes utilized in Brazilian breeding programs revealed clustering according to sugar, phenolic acid, and flavone contents. PMID:27152527

  19. Transcript and metabolite alterations increase ganoderic acid content in Ganoderma lucidum using acetic acid as an inducer.

    Ren, Ang; Li, Xiong-Biao; Miao, Zhi-Gang; Shi, Liang; Jaing, Ai-Liang; Zhao, Ming-Wen

    2014-12-01

    Acetic acid at 5-8 mM increased ganoderic acid (GA) accumulation in Ganoderma lucidum. After optimization by the response surface methodology, the GA content reached 5.5/100 mg dry weight, an increase of 105% compared with the control. The intermediate metabolites of GA biosynthesis, lanosterol and squalene also increased to 47 and 15.8 μg/g dry weight, respectively, in response to acetic acid. Acetic acid significantly induced transcription levels of sqs, lano, hmgs and cyp51 in the GA biosynthesis pathway. An acetic acid-unregulated acetyl coenzyme A synthase (acs) gene was selected from ten candidate homologous acs genes. The results indicate that acetic acid alters the expression of genes related to acetic acid assimilation and increases GA biosynthesis and the metabolic levels of lanosterol, squalene and GA-a, thereby resulting in GA accumulation. PMID:25216642

  20. Identification of mebeverine acid as the main circulating metabolite of mebeverine in man.

    Stockis, Arnel; Guelen, P J M; de Vos, D

    2002-06-20

    The intestinal spasmolytic drug mebeverine is known to undergo fast in vivo enzymatic hydrolysis into mebeverine alcohol and veratric acid. A reversed-phase HPLC method with coulometric detection was developed in order to assay the hitherto unidentified secondary metabolite mebeverine acid. After intake of a single oral dose of 405 mg mebeverine hydrochloride in four healthy human volunteers, peak plasma concentrations of mebeverine acid were found to be 1000-fold higher than those of mebeverine alcohol, i.e. approximately 3 microg/ml versus 3 ng/ml. The appearance of mebeverine acid in plasma (median T(max)=1.25 h) as well as its disappearance (median apparent t(1/2)=1.1 h) were rapid. The urinary excretion of mebeverine acid within the first 4 h after dosing amounted to 67% of the mebeverine dose (median range: 23-107%). Mebeverine acid appears to be a valuable marker of oral exposure to mebeverine. PMID:12062694

  1. Glucuronic acid and the ethanol metabolite ethyl-glucuronide cause Toll-like receptor 4 activation and enhanced pain

    Lewis, Susannah S.; Hutchinson, Mark R; Zhang, Yingning; Hund, Dana K.; Maier, Steven F.; Rice, Kenner C.; Watkins, Linda R

    2013-01-01

    We have previously observed that the non-opioid morphine metabolite, morphine-3-glucuronide, enhances pain via a toll-like receptor 4 (TLR4) dependent mechanism. The present studies were undertaken to determine whether TLR4-dependent pain enhancement generalizes to other classes of glucuronide metabolites. In silico modeling predicted that glucuronic acid alone and ethyl glucuronide, a minor but long-lasting ethanol metabolite, would dock to the same MD-2 portion of the TLR4 receptor complex ...

  2. The host metabolite D-serine contributes to bacterial niche specificity through gene selection

    Connolly, James P. R.; Goldstone, Robert J.; Burgess, Karl; Cogdell, Richard J.; Beatson, Scott A.; Vollmer, Waldemar; Smith, David G. E.; Roe, Andrew

    2015-01-01

    Escherichia coli comprise a diverse array of both commensals and niche-specific pathotypes. The ability to cause disease results from both carriage of specific virulence factors and regulatory control of these via environmental stimuli. Moreover, host metabolites further refine the response of bacteria to their environment and can dramatically affect the outcome of the host–pathogen interaction. Here, we demonstrate that the host metabolite, D-serine, selectively affects gene expression in E....

  3. Association between Circulating Vitamin D Metabolites and Fecal Bile Acid Concentrations.

    Jacobs, Elizabeth T; Haussler, Mark R; Alberts, David S; Kohler, Lindsay N; Lance, Peter; Martínez, María Elena; Roe, Denise J; Jurutka, Peter W

    2016-07-01

    Although hydrophobic bile acids have been demonstrated to exhibit cytotoxic and carcinogenic effects in the colorectum, ursodeoxycholic acid (UDCA) has been investigated as a potential chemopreventive agent. Vitamin D has been shown to play a role in both bile acid metabolism and in the development of colorectal neoplasia. Using a cross-sectional design, we sought to determine whether baseline circulating concentrations of the vitamin D metabolites 25(OH)D and 1,25(OH)2D were associated with baseline fecal bile acid concentrations in a trial of UDCA for the prevention of colorectal adenoma recurrence. We also prospectively evaluated whether vitamin D metabolite concentrations modified the effect of UDCA on adenoma recurrence. After adjustment for age, sex, BMI, physical activity, and calcium intake, adequate concentrations of 25(OH)D (≥30 ng/mL) were statistically significantly associated with reduced odds for high levels of total [OR, 0.61; 95% confidence interval (CI), 0.38-0.97], and primary (OR, 0.61; 95% CI, 0.38-0.96) bile acids, as well as individually with chenodeoxycholic acid (OR, 0.39; 95% CI, 0.24-0.63) and cholic acid (OR, 0.56; 95% CI, 0.36-0.90). No significant associations were observed for 1,25(OH)2D and high versus low fecal bile acid concentrations. In addition, neither 25(OH)D nor 1,25(OH)2D modified the effect of UDCA on colorectal adenoma recurrence. In conclusion, this is the first study to demonstrate an inverse relationship between circulating levels of 25(OH)D and primary fecal bile acid concentrations. These results support prior data demonstrating that vitamin D plays a key role in bile acid metabolism, and suggest a potential mechanism of action for 25(OH)D in colorectal cancer prevention. Cancer Prev Res; 9(7); 589-97. ©2016 AACR. PMID:27138789

  4. Usnic acid, a secondary metabolite of lichens and its effect on in vitro digestibility in reindeer

    R. Thomas Palo

    1993-10-01

    Full Text Available Usnic acid, a common secondary metabolite in prefered lichens by reindeer and caribou, has been tested for its effect on In Vitro Dry Matter Digestibility (IVDMD using inocula from four reindeer. When Cladonia alpestris (stellaris (OpicJ was used as substrate and reindeer rumen liqour as media of incubation together with usnic acid, digestibility was considerably enhanced. This was also true for a lower prefered lichen Stereocaulon paschale (L., but the effect was less pronounced. The results suggest that reindeer host some rumen microorganism able to metabolize lichen secondary metabolities.

  5. Fatty acid amide hydrolase-dependent generation of antinociceptive drug metabolites acting on TRPV1 in the brain.

    David A Barrière

    Full Text Available The discovery that paracetamol is metabolized to the potent TRPV1 activator N-(4-hydroxyphenyl-5Z,8Z,11Z,14Z-eicosatetraenamide (AM404 and that this metabolite contributes to paracetamol's antinociceptive effect in rodents via activation of TRPV1 in the central nervous system (CNS has provided a potential strategy for developing novel analgesics. Here we validated this strategy by examining the metabolism and antinociceptive activity of the de-acetylated paracetamol metabolite 4-aminophenol and 4-hydroxy-3-methoxybenzylamine (HMBA, both of which may undergo a fatty acid amide hydrolase (FAAH-dependent biotransformation to potent TRPV1 activators in the brain. Systemic administration of 4-aminophenol and HMBA led to a dose-dependent formation of AM404 plus N-(4-hydroxyphenyl-9Z-octadecenamide (HPODA and arvanil plus olvanil in the mouse brain, respectively. The order of potency of these lipid metabolites as TRPV1 activators was arvanil = olvanil>>AM404> HPODA. Both 4-aminophenol and HMBA displayed antinociceptive activity in various rodent pain tests. The formation of AM404, arvanil and olvanil, but not HPODA, and the antinociceptive effects of 4-aminophenol and HMBA were substantially reduced or disappeared in FAAH null mice. The activity of 4-aminophenol in the mouse formalin, von Frey and tail immersion tests was also lost in TRPV1 null mice. Intracerebroventricular injection of the TRPV1 blocker capsazepine eliminated the antinociceptive effects of 4-aminophenol and HMBA in the mouse formalin test. In the rat, pharmacological inhibition of FAAH, TRPV1, cannabinoid CB1 receptors and spinal 5-HT3 or 5-HT1A receptors, and chemical deletion of bulbospinal serotonergic pathways prevented the antinociceptive action of 4-aminophenol. Thus, the pharmacological profile of 4-aminophenol was identical to that previously reported for paracetamol, supporting our suggestion that this drug metabolite contributes to paracetamol's analgesic activity via

  6. Fatty Acid Amide Hydrolase-Dependent Generation of Antinociceptive Drug Metabolites Acting on TRPV1 in the Brain

    Blomgren, Anders; Simonsen, Charlotte; Daulhac, Laurence; Libert, Frédéric; Chapuy, Eric; Etienne, Monique; Högestätt, Edward D.; Zygmunt, Peter M.; Eschalier, Alain

    2013-01-01

    The discovery that paracetamol is metabolized to the potent TRPV1 activator N-(4-hydroxyphenyl)-5Z,8Z,11Z,14Z-eicosatetraenamide (AM404) and that this metabolite contributes to paracetamol’s antinociceptive effect in rodents via activation of TRPV1 in the central nervous system (CNS) has provided a potential strategy for developing novel analgesics. Here we validated this strategy by examining the metabolism and antinociceptive activity of the de-acetylated paracetamol metabolite 4-aminophenol and 4-hydroxy-3-methoxybenzylamine (HMBA), both of which may undergo a fatty acid amide hydrolase (FAAH)-dependent biotransformation to potent TRPV1 activators in the brain. Systemic administration of 4-aminophenol and HMBA led to a dose-dependent formation of AM404 plus N-(4-hydroxyphenyl)-9Z-octadecenamide (HPODA) and arvanil plus olvanil in the mouse brain, respectively. The order of potency of these lipid metabolites as TRPV1 activators was arvanil = olvanil>>AM404> HPODA. Both 4-aminophenol and HMBA displayed antinociceptive activity in various rodent pain tests. The formation of AM404, arvanil and olvanil, but not HPODA, and the antinociceptive effects of 4-aminophenol and HMBA were substantially reduced or disappeared in FAAH null mice. The activity of 4-aminophenol in the mouse formalin, von Frey and tail immersion tests was also lost in TRPV1 null mice. Intracerebroventricular injection of the TRPV1 blocker capsazepine eliminated the antinociceptive effects of 4-aminophenol and HMBA in the mouse formalin test. In the rat, pharmacological inhibition of FAAH, TRPV1, cannabinoid CB1 receptors and spinal 5-HT3 or 5-HT1A receptors, and chemical deletion of bulbospinal serotonergic pathways prevented the antinociceptive action of 4-aminophenol. Thus, the pharmacological profile of 4-aminophenol was identical to that previously reported for paracetamol, supporting our suggestion that this drug metabolite contributes to paracetamol’s analgesic activity via activation

  7. Occurrence and fate of the human pharmaceutical metabolite ritalinic acid in the aquatic system.

    Letzel, Marion; Weiss, Klaus; Schüssler, Walter; Sengl, Manfred

    2010-12-01

    To investigate the occurrence and fate of ritalinic acid - the main human metabolite of the psychostimulant drug methylphenidate - in the aquatic environment, a HPLC-electrospray-MS/MS method for the quantification of ritalinic acid in wastewater, surface water and bank filtrate was developed. Carbamazepine known as very stable in the aquatic environment was analyzed as anthropogenic marker in parallel. Furthermore, the removal of ritalinic acid was studied in a sewage treatment plant using an activated sludge system during a field study and in lab-scale plants. In good agreement between lab-scale and field studies a low removal rate of 13% and 23%, respectively, was determined. As a consequence, the concentration of ritalinic acid in the wastewater effluents were in the range of bank filtrate samples in 100-850 m distance from the river up to 5 ngL(-1) demonstrating the widespread occurrence of this stable metabolite in the aquatic environment. A comparison to available sales data shows that a significant amount of methylphenidate applied can be found in waters as ritalinic acid. PMID:20932550

  8. Ion-pair chromatography of acidic drug metabolites and endogenic compounds.

    Fransson, B; Wahlund, K G; Johansson, I M; Schill, G

    1976-09-29

    Liquid-liquid chromatographic systems based on ion-pair partition with silica microparticles as the support for the stationary phase have been used for the separation of anionic compounds of biochemical and pharmacological interest. A high separating efficiency can be obtained with both aqueous and organic mobile phases and the retention is easily regulated by the nature and the concentration of the quaternary ammonium counter ion, present in the aqueous phase. The influence of the composition of the liquid phases on the selectivity and separating efficiency has been studied, as well as equilibration methods and the stability of the systems. Examples are given of separations of sulphonamides, barbiturates, glucuronic and sulphuric acid conjugates of steroidal compounds and phenols glycine conjugates of carboxylic acids (hippuric, nicotinuric and salicyluric acid) and anionic metabolites of biogenic amines (indoleacetic, benzoic, mandelic and phenylacetic acid derivatives). PMID:10314

  9. Amino Acid and Secondary Metabolite Production in Embryogenic and Non-Embryogenic Callus of Fingerroot Ginger (Boesenbergia rotunda.

    Theresa Lee Mei Ng

    Full Text Available Interest in the medicinal properties of secondary metabolites of Boesenbergia rotunda (fingerroot ginger has led to investigations into tissue culture of this plant. In this study, we profiled its primary and secondary metabolites, as well as hormones of embryogenic and non-embryogenic (dry and watery callus and shoot base, Ultra Performance Liquid Chromatography-Mass Spectrometry together with histological characterization. Metabolite profiling showed relatively higher levels of glutamine, arginine and lysine in embryogenic callus than in dry and watery calli, while shoot base tissue showed an intermediate level of primary metabolites. For the five secondary metabolites analyzed (ie. panduratin, pinocembrin, pinostrobin, cardamonin and alpinetin, shoot base had the highest concentrations, followed by watery, dry and embryogenic calli. Furthermore, intracellular auxin levels were found to decrease from dry to watery calli, followed by shoot base and finally embryogenic calli. Our morphological observations showed the presence of fibrils on the cell surface of embryogenic callus while diphenylboric acid 2-aminoethylester staining indicated the presence of flavonoids in both dry and embryogenic calli. Periodic acid-Schiff staining showed that shoot base and dry and embryogenic calli contained starch reserves while none were found in watery callus. This study identified several primary metabolites that could be used as markers of embryogenic cells in B. rotunda, while secondary metabolite analysis indicated that biosynthesis pathways of these important metabolites may not be active in callus and embryogenic tissue.

  10. Amino Acid and Secondary Metabolite Production in Embryogenic and Non-Embryogenic Callus of Fingerroot Ginger (Boesenbergia rotunda).

    Ng, Theresa Lee Mei; Karim, Rezaul; Tan, Yew Seong; Teh, Huey Fang; Danial, Asma Dazni; Ho, Li Sim; Khalid, Norzulaani; Appleton, David Ross; Harikrishna, Jennifer Ann

    2016-01-01

    Interest in the medicinal properties of secondary metabolites of Boesenbergia rotunda (fingerroot ginger) has led to investigations into tissue culture of this plant. In this study, we profiled its primary and secondary metabolites, as well as hormones of embryogenic and non-embryogenic (dry and watery) callus and shoot base, Ultra Performance Liquid Chromatography-Mass Spectrometry together with histological characterization. Metabolite profiling showed relatively higher levels of glutamine, arginine and lysine in embryogenic callus than in dry and watery calli, while shoot base tissue showed an intermediate level of primary metabolites. For the five secondary metabolites analyzed (ie. panduratin, pinocembrin, pinostrobin, cardamonin and alpinetin), shoot base had the highest concentrations, followed by watery, dry and embryogenic calli. Furthermore, intracellular auxin levels were found to decrease from dry to watery calli, followed by shoot base and finally embryogenic calli. Our morphological observations showed the presence of fibrils on the cell surface of embryogenic callus while diphenylboric acid 2-aminoethylester staining indicated the presence of flavonoids in both dry and embryogenic calli. Periodic acid-Schiff staining showed that shoot base and dry and embryogenic calli contained starch reserves while none were found in watery callus. This study identified several primary metabolites that could be used as markers of embryogenic cells in B. rotunda, while secondary metabolite analysis indicated that biosynthesis pathways of these important metabolites may not be active in callus and embryogenic tissue. PMID:27258536

  11. Decrease of intracellular pH as possible mechanism of embryotoxicity of glycol ether alkoxyacetic acid metabolites

    Embryotoxicity of glycol ethers is caused by their alkoxyacetic acid metabolites, but the mechanism underlying the embryotoxicity of these acid metabolites is so far not known. The present study investigates a possible mechanism underlying the embryotoxicity of glycol ether alkoxyacetic acid metabolites using the methoxyacetic acid (MAA) metabolite of ethylene glycol monomethyl ether as the model compound. The results obtained demonstrate an MAA-induced decrease of the intracellular pH (pHi) of embryonic BALB/c-3T3 cells as well as of embryonic stem (ES)-D3 cells, at concentrations that affect ES-D3 cell differentiation. These results suggest a mechanism for MAA-mediated embryotoxicity similar to the mechanism of embryotoxicity of the drugs valproic acid and acetazolamide (ACZ), known to decrease the pHiin vivo, and therefore used as positive controls. The embryotoxic alkoxyacetic acid metabolites ethoxyacetic acid, butoxyacetic acid and phenoxyacetic acid also caused an intracellular acidification of BALB/c-3T3 cells at concentrations that are known to inhibit ES-D3 cell differentiation. Two other embryotoxic compounds, all-trans-retinoic acid and 5-fluorouracil, did not decrease the pHi of embryonic cells at concentrations that affect ES-D3 cell differentiation, pointing at a different mechanism of embryotoxicity of these compounds. MAA and ACZ induced a concentration-dependent inhibition of ES-D3 cell differentiation, which was enhanced by amiloride, an inhibitor of the Na+/H+-antiporter, corroborating an important role of the pHi in the embryotoxic mechanism of both compounds. Together, the results presented indicate that a decrease of the pHi may be the mechanism of embryotoxicity of the alkoxyacetic acid metabolites of the glycol ethers.

  12. Biosynthesis of a mannolipid containing a metabolite of retinoic acid by 3T12 mouse fibroblasts

    Retinol and retinoic acid (RA) increase the adhesive properties of spontaneously transformed mouse fibroblasts and the incorporation of [2-3H]mannose into cellular glycoconjugates. Therefore we searched for a mannolipid of retinoic acid similar to mannosylretinylphosphate (MRP) in these cells. Labeled RA was incorporated into a compound similar to standard MRP. This metabolite contained the same 3H:14C ratio as the precursor [11, 12-3H, 15-14C]retinoic acid, demonstrating that no decarboxylation had occurred. A doubly labeled mannolipid was obtained from cells incubated with [2-3H]mannose and [15-14C]retinoic acid. This mannolipid was readily cleaved by mild acid, yielding [3H]mannosephosphate and a compound that migrated as standard anhydroretinol at Rf 0.93. Standard all trans-MRP yields all-trans-anhydroretinol under these conditions. A HPLC system was developed to further characterize the mannolipids obtained from retinol and retinoic acid in 3T12 cells. [15-3H]Retinol and [15-14C]retinoic acid were incorporated into mannolipids that cochromatographed with standard MRP. The mixture of the [15-3H]retinol and [15-14C]retinoic acid derived mannolipids was subjected to mild acid hydrolysis, after purification by HPLC yielding all-trans-[3H]anhydroretinol and a [14C]labeled product which was eluted from HPLC as a slightly more polar compound than all-trans-anhydroretinol. The retinoic acid-derived mannolipid (MXP) represented approximately 4% of the total radioactivity in the methanolic extract of 3T12 cells incubated in labeled retinoic acid. However, if the cells were incubated for an additional 20 hours in the absence of the radioactive precursor, MXP represented 40% of the total extracted radioactivity

  13. Synthesis and stability study of a new major metabolite of gamma-hydroxybutyric acid

    Petersen, Ida Nymann; Kristensen, Jesper Langgaard; Tortzen, Christian;

    2013-01-01

    ¿-Hydroxybutanoic acid (GHB) is used as a date-rape drug, which renders the victims unconscious and defenceless. Intoxications are very difficult to detect for forensic scientists due to rapid metabolism to endogenous levels of GHB. We recently discovered a new major metabolite, 2, of GHB (1......, we have assessed the stability of GHB glucuronide 2 by mimicking the natural pH range for urine, which is of importance in the development of new analytical methods. Using NMR we show that GHB glucuronide 2 is highly stable towards aqueous hydrolysis within the pH range normally observed for urine...

  14. Cytochrome P450-generated metabolites derived from ω-3 fatty acids attenuate neovascularization

    Yanai, Ryoji; Mulki, Lama; Hasegawa, Eiichi; Takeuchi, Kimio; Sweigard, Harry; Suzuki, Jun; Gaissert, Philipp; Vavvas, Demetrios G.; Sonoda, Koh-Hei; Rothe, Michael; Schunck, Wolf-Hagen; Miller, Joan W.; Connor, Kip M.

    2014-01-01

    The ω-3 long-chain polyunsaturated fatty acids are a class of dietary lipids that are highly enriched in the central nervous system and the retina. We demonstrate that dietary enrichment with ω-3s suppresses choroidal neovascularization in a mouse model of age-related macular degeneration (AMD), a leading cause of blindness. The ω-3s have anti-inflammatory properties and compete with ω-6s for downstream lipid metabolite synthesis at the cytochrome P450 (CYP) level. Specifically, 17,18- epoxye...

  15. Borna disease virus infection perturbs energy metabolites and amino acids in cultured human oligodendroglia cells.

    Rongzhong Huang

    Full Text Available BACKGROUND: Borna disease virus is a neurotropic, non-cytolytic virus that has been widely employed in neuroscientific research. Previous studies have revealed that metabolic perturbations are associated with Borna disease viral infection. However, the pathophysiological mechanism underlying its mode of action remains unclear. METHODOLOGY: Human oligodendroglia cells infected with the human strain Borna disease virus Hu-H1 and non-infected matched control cells were cultured in vitro. At day 14 post-infection, a proton nuclear magnetic resonance-based metabonomic approach was used to differentiate the metabonomic profiles of 28 independent intracellular samples from Borna disease virus-infected cells (n = 14 and matched control cells (n = 14. Partial least squares discriminant analysis was performed to demonstrate that the whole metabonomic patterns enabled discrimination between the two groups, and further statistical testing was applied to determine which individual metabolites displayed significant differences between the two groups. FINDINGS: Metabonomic profiling revealed perturbations in 23 metabolites, 19 of which were deemed individually significant: nine energy metabolites (α-glucose, acetate, choline, creatine, formate, myo-inositol, nicotinamide adenine dinucleotide, pyruvate, succinate and ten amino acids (aspartate, glutamate, glutamine, glycine, histidine, isoleucine, phenylalanine, threonine, tyrosine, valine. Partial least squares discriminant analysis demonstrated that the whole metabolic patterns enabled statistical discrimination between the two groups. CONCLUSION: Borna disease viral infection perturbs the metabonomic profiles of several metabolites in human oligodendroglia cells cultured in vitro. The findings suggest that Borna disease virus manipulates the host cell's metabolic network to support viral replication and proliferation.

  16. Contribution to the study of radioisotopic methods in pharmacokinetics. Application to specific determinations of drugs or their metabolites

    The aim of this work was to refute one of the major criticisms expressed on the used of labelled molecules, that they give an overall result. Techniques were therefore developed to determine quantitatively and specifically the kinetics of the drug itself or its metabolites. Two methods turning to account the great sensitivity and facility offered by labelled molecules have been adopted: - reverse isotopic dilution and double isotopic dilution, applied to some medicinal molecules. In part one the glipentide labelled molecule was used to measure the unchanged product in rat plasma: the kinetics are established. In part two the plasma fraction curves of unchanged products and their metabolites were studied for two molecules of similar structure: cyclobutane carboxylic acid and propyl-3 cyclobutane carboxylic acid. Finally a radiocompetitive method to determine a sulfamido-benzoic diuretic, based on the interaction with carbonic anhydrase, was investigated. The sensitivity of these radioisotopic methods depends on the specific activity of the labelled molecule. For the glipentide for instance, where the specific activity is very high, as little as 2 ng/ml of plasma can be determined. The specific activities of cyclobutane carboxylic, propyl-3 cyclobutane carboxylic and sulfamido-3 chloro-4 benzoic acids are not high enough for measurements better than 1 μg/ml plasma to be obtained

  17. Role of omega-3 fatty acids and their metabolites in asthma and allergic diseases

    Jun Miyata

    2015-01-01

    Full Text Available Omega-3 fatty acids, docosahexaenoic acid (DHA and eicosapentaenoic acid (EPA, are found naturally in fish oil and are commonly thought to be anti-inflammatory nutrients, with protective effects in inflammatory diseases including asthma and allergies. The mechanisms of these effects remain mostly unknown but are of great interest for their potential therapeutic applications. Large numbers of epidemiological and observational studies investigating the effect of fish intake or omega-3 fatty acid supplementation during pregnancy, lactation, infancy, childhood, and adulthood on asthmatic and allergic outcomes have been conducted. They mostly indicate protective effects and suggest a causal relationship between decreased intake of fish oil in modernized diets and an increasing number of individuals with asthma or other allergic diseases. Specialized pro-resolving mediators (SPM: protectins, resolvins, and maresins are generated from omega-3 fatty acids such as EPA and DHA via several enzymatic reactions. These mediators counter-regulate airway eosinophilic inflammation and promote the resolution of inflammation in vivo. Several reports have indicated that the biosynthesis of SPM is impaired, especially in severe asthma, which suggests that chronic inflammation in the lung might result from a resolution defect. This article focuses on the beneficial aspects of omega-3 fatty acids and offers recent insights into their bioactive metabolites including resolvins and protectins.

  18. Prevalence and impact of single-strain starter cultures of lactic acid bacteria on metabolite formation in sourdough.

    Ravyts, Frédéric; De Vuyst, Luc

    2011-09-01

    Flavour of type II sourdoughs is influenced by the ingredients, processing conditions, and starter culture composition. It is, however, not fully clear to what extent different sourdough lactic acid bacteria (LAB) contribute to flavour. Therefore, two types of flour (rye and wheat) and different LAB starter culture strains were used to prepare sourdoughs, thereby leaving the yeast microbiota uncontrolled. All LAB starter culture strains tested were shown to be prevalent and to acidify the flour/water mixture to pH values between 3.1 and 3.9 after 24h of fermentation. Multiple aldehydes, alcohols, ketones, and carboxylic acids were produced by the sourdough-associated microbiota throughout the fermentation period. Based on the organoleptic evaluation of breads produced with these sourdoughs, five LAB strains were selected to perform prolonged wheat and rye fermentations as to their capacity to result in an acidic (Lactobacillus fermentum IMDO 130101, Lactobacillus plantarum IMDO 130201, and Lactobacillus crustorum LMG 23699), buttermilk-like (Lactobacillus amylovorus DCE 471), or fruity flavour (Lactobacillus sakei CG1). Upon prolonged fermentation, higher metabolite concentrations were produced. For instance, L. sakei CG1 produced the highest amounts of 3-methyl-1-butanol, which was further converted into 3-methylbutyl acetate. The latter compound resulted in a fruity banana flavour after 48h of fermentation, probably due to yeast interference. Rye fermentations resulted in sourdoughs richer in volatiles than wheat, including 3-methyl-1-butanol, 2-phenylethanol, and ethyl acetate. PMID:21645811

  19. Secondary metabolites from the sponge Tedania anhelans: Isolation and characterization of two novel pyrazole acids and other metabolites

    Parameswaran, P.S.; Naik, C.G.; Hegde, V.R.

    Chemical investigation of the methanol extract of the sponge Tedania anhelans yielded the two unusual heteroaromatic acids, pyrazole-3(5)-carboxylic acid (2) and 4-methylpyrazole-3(5)-carboxylic acid (3), which are reported for the first time...

  20. Lichen secondary metabolite evernic acid as potential quorum sensing inhibitor against Pseudomonas aeruginosa.

    Gökalsın, Barış; Sesal, Nüzhet Cenk

    2016-09-01

    Cystic Fibrosis is a genetic disease and it affects the respiratory and digestive systems. Pseudomonas aeruginosa infections in Cystic Fibrosis are presented as the main cause for high mortality and morbidity rates. Pseudomonas aeruginosa populations can regulate their virulence gene expressions via the bacterial communication system: quorum sensing. Inhibition of quorum sensing by employing quorum sensing inhibitors can leave the bacteria vulnerable. Therefore, determining natural sources to obtain potential quorum sensing inhibitors is essential. Lichens have ethnobotanical value for their medicinal properties and it is possible that their secondary metabolites have quorum sensing inhibitor properties. This study aims to investigate an alternative treatment approach by utilizing lichen secondary metabolite evernic acid to reduce the expressions of Pseudomonas aeruginosa virulence factors by inhibiting quorum sensing. For this purpose, fluorescent monitor strains were utilized for quorum sensing inhibitor screens and quantitative reverse-transcriptase PCR analyses were conducted for comparison. Results indicate that evernic acid is capable of inhibiting Pseudomonas aeruginosa quorum sensing systems. PMID:27465850

  1. Protective Effects of Dihydrocaffeic Acid, a Coffee Component Metabolite, on a Focal Cerebral Ischemia Rat Model

    Kyungjin Lee

    2015-06-01

    Full Text Available We recently reported the protective effects of chlorogenic acid (CGA in a transient middle cerebral artery occlusion (tMCAo rat model. The current study further investigated the protective effects of the metabolites of CGA and dihydrocaffeic acid (DHCA was selected for further study after screening using the same tMCAo rat model. In the current study, tMCAo rats (2 h of MCAo followed by 22 h of reperfusion were injected with various doses of DHCA at 0 and 2 h after onset of ischemia. We assessed brain damage, functional deficits, brain edema, and blood-brain barrier damage at 24 h after ischemia. For investigating the mechanism, in vitro zymography and western blotting analysis were performed to determine the expression and activation of matrix metalloproteinase (MMP-2 and -9. DHCA (3, 10, and 30 mg/kg, i.p. dose-dependently reduced brain infarct volume, behavioral deficits, brain water content, and Evans Blue (EB leakage. DHCA inhibited expression and activation of MMP-2 and MMP-9. Therefore, DHCA might be one of the important metabolites of CGA and of natural products, including coffee, with protective effects on ischemia-induced neuronal damage and brain edema.

  2. Total synthesis of the aristolochic acids, their major metabolites, and related compounds.

    Attaluri, Sivaprasad; Iden, Charles R; Bonala, Radha R; Johnson, Francis

    2014-07-21

    Plants from the Aristolochia genus have been recommended for the treatment of a variety of human ailments since the time of Hippocrates. However, many species produce the highly toxic aristolochic acids (AAs), which are both nephrotoxic and carcinogenic. For the purposes of extensive biological studies, a versatile approach to the synthesis of the AAs and their major metabolites was devised based primarily on a Suzuki-Miyaura coupling reaction. The key to success lies in the preparation of a common ring-A precursor, namely, the tetrahydropyranyl ether of 2-nitromethyl-3-iodo-4,5-methylendioxybenzyl alcohol (27), which was generated in excellent yield by oxidation of the aldoxime precursor 26. Suzuki-Miyaura coupling of 27 with a variety of benzaldehyde 2-boronates was accompanied by an aldol condensation/elimination reaction to give the desired phenanthrene intermediate directly. Deprotection of the benzyl alcohol followed by two sequential oxidation steps gave the desired phenanthrene nitrocarboxylic acids. This approach was used to synthesize AAs I-IV and several other related compounds, including AA I and AA II bearing an aminopropyloxy group at position-6, which were required for further conversion to fluorescent biological probes. Further successful application of the Suzuki-Miyaura coupling reaction to the synthesis of the N-hydroxyaristolactams of AA I and AA II then allowed the synthesis of the putative, but until now elusive, N-acetoxy- and N-sulfonyloxy-aristolactam metabolites. PMID:24877584

  3. Potential of selected lactic acid bacteria to produce food compatible antifungal metabolites.

    De Muynck, Cassandra; Leroy, Annelies I J; De Maeseneire, Sofie; Arnaut, Filip; Soetaert, Wim; Vandamme, Erick J

    2004-01-01

    The aim of this study was to assess the potential of lactic acid bacteria to inhibit the outgrowth of some common food-spoiling fungi. Culture supernatants of 17 Lactic acid bacterial strains as well as of three commercial probiotic cultures were evaluated for antifungal activity using an agar-diffusion method. The method parameters were chosen in order to reveal compounds for potential use in food (bio)preservation. Thirteen strains showed antifungal activity of which five strains were very promising: Lactobacillus acidophilus LMG 9433, L. amylovorus DSM 20532, L. brevis LMG 6906, L. coryniformis subsp. coryniformis LMG 9196 and L. plantarum LMG 6907. Four of these five strains were further examined; it was found that the produced antifungal metabolites were pH-dependent. The exact chemical nature of these substances has not been revealed yet. PMID:15646380

  4. Hypoxia optimises tumour growth by controlling nutrient import and acidic metabolite export.

    Parks, Scott K; Cormerais, Yann; Marchiq, Ibtissam; Pouyssegur, Jacques

    2016-01-01

    In their quest for survival and successful growth, cancer cells optimise their cellular processes to enable them to outcompete normal cells in their microenvironment. In essence cancer cells: (i) enhance uptake of nutrients/metabolites, (ii) utilise nutrients more efficiently via metabolic alterations and (iii) deal with the metabolic waste products in a way that furthers their progression while hampering the survival of normal tissue. Hypoxia Inducible Factors (HIFs) act as essential drivers of these adaptations via the promotion of numerous membrane proteins including glucose transporters (GLUTs), monocarboxylate transporters (MCTs), amino-acid transporters (LAT1, xCT), and acid-base regulating carbonic anhydrases (CAs). In addition to a competitive growth advantage for tumour cells, these HIF-regulated proteins are implicated in metastasis, cancer 'stemness' and the immune response. Current research indicates that combined targeting of these HIF-regulated membrane proteins in tumour cells will provide promising therapeutic strategies in the future. PMID:26724171

  5. Fatty Acid Amide Hydrolase-Dependent Generation of Antinociceptive Drug Metabolites Acting on TRPV1 in the Brain

    Barriere, David A.; Mallet, Christophe; Blomgren, Anders; Simonsen, Charlotte; Daulhac, Laurence; Libert, Frederic; Chapuy, Eric; Etienne, Monique; Högestätt, Edward; Zygmunt, Peter; Eschalier, Alain

    2013-01-01

    The discovery that paracetamol is metabolized to the potent TRPV1 activator N-(4-hydroxyphenyl)-5Z, 8Z, 11Z, 14Z-eicosatetraenamide (AM404) and that this metabolite contributes to paracetamol's antinociceptive effect in rodents via activation of TRPV1 in the central nervous system (CNS) has provided a potential strategy for developing novel analgesics. Here we validated this strategy by examining the metabolism and antinociceptive activity of the de-acetylated paracetamol metabolite 4-ami...

  6. Plasma concentrations of amino acid and nicotinamide metabolites in rheumatoid arthritis - potential biomarkers of disease activity and drug treatment.

    Smolenska, Zaneta; Smolenski, Ryszard T; Zdrojewski, Zbigniew

    2016-05-01

    This study aimed to evaluate changes in plasma amino acid and nicotinamide metabolites concentrations in rheumatoid arthritis (RA) in a search for potential biomarkers of the disease activity and the effect treatment. Analysis of plasma metabolite patterns with liquid chromatography/mass spectrometry revealed specific changes in RA as well as correlations with clinical parameters. Combined concentration parameter calculated as [aspartic acid] + [threonine] + [tryptophan] - [histidine] - [phenylalanine] offered the strongest correlation (p < 0.001) with pain joint count, swollen joint count and DAS 28. Such analysis of amino acid and related metabolite pattern offers potential for diagnosis as well as for monitoring disease progression and therapy in RA. PMID:26811910

  7. Plasma cathepsin D isoforms and their active metabolites increase after myocardial infarction and contribute to plasma renin activity.

    Naseem, R Haris; Hedegard, Wade; Henry, Timothy D; Lessard, Jennifer; Sutter, Kathryn; Katz, Stephen A

    2005-03-01

    Plasma renin activity (PRA) is often found to increase after myocardial infarction (MI). Elevated PRA may contribute to increased myocardial angiotensin II that is responsible for maladaptive remodeling of the myocardium after MI. We hypothesized that MI would also result in cardiac release of cathepsin D, a ubiquitous lysosomal enzyme with high renin sequence homology. Cathepsin D release from damaged myocardial tissue could contribute to angiotensin formation by acting as an enzymatic alternate to renin. We assessed circulating renin and cathepsin D from both control and MI patient plasma (7-20 hours after MI) using shallow gradient focusing that allowed for independent measurement of both enzymes. Cathepsin D was increased significantly in the plasma after MI (P < 0.001). Furthermore, circulating active cathepsin D metabolites were also significantly elevated after MI (P < 0.04), and contained the majority of cathepsin D activity in plasma. Spiking control plasma with cathepsin D resulted in a variable but significant (P = 0.005) increase in PRA using a clinical assay. We conclude that 7-20 hours after MI, plasma cathepsin D is significantly elevated and most of the active enzymatic activity is circulating as plasma metabolites. Circulating cathepsin D can falsely increase clinical PRA determinations, and may also provide an alternative angiotensin formation pathway after MI. PMID:15739123

  8. Enantioselective separation and transformation of metalaxyl and its major metabolite metalaxyl acid in tomato and cucumber.

    Li, Yuanbo; Dong, Fengshou; Liu, Xingang; Xu, Jun; Chen, Xiu; Han, Yongtao; Cheng, Youpu; Jian, Qiu; Zheng, Yongquan

    2013-11-01

    Many studies have shown the enantioselective transformation of metalaxyl in soil and animals, but little is known about the enantioselective biotransformation of metalaxyl in plants. In this study, a sensitive and convenient chiral liquid chromatography-tandem mass spectrometry method was developed for simultaneous measurement of the enantiomers of metalaxyl and its major metabolite, metalaxyl acid, in tomato and cucumber. Separation was by reversed-phase chromatography on a Chiralcel OD-RH column under isocratic conditions using acetonitrile-water (60/40, v/v) as mobile phase. The proposed method was successfully applied to investigate the possible enantioselective transformation of metalaxyl as well as the formation of metalaxyl acid in tomato and cucumber. In both vegetables, the (-)-R-enantiomer was preferentially degraded, resulting in relative enrichment of the (+)-S-enantiomer. Furthermore, formation of the metalaxyl acid enantiomers was also shown to be enantioselective, with the R-metalaxyl acid formed at a faster rate. This is the first evidence of enantioselective transformation of metalaxyl in vegetables, and the results should be considered in future environmental risk and food safety evaluations. PMID:23768319

  9. THREE MICROSOMAL OMEGA-3 FATTY ACID DESATURASE GENES CONTRIBUTE TO SOYBEAN LINOLENIC ACID LEVELS

    Three independent genetic loci have been shown to contribute to soybean (Glycine max L.) seed linolenic acid levels, including the well-characterized Fan locus. Linolenic acid is the product of omega-3-fatty acid desaturase enzyme activity. The objective of this study was to identify and character...

  10. Synthesis and stability study of a new major metabolite of γ-hydroxybutyric acid

    Ida Nymann Petersen

    2013-04-01

    Full Text Available γ-Hydroxybutanoic acid (GHB is used as a date-rape drug, which renders the victims unconscious and defenceless. Intoxications are very difficult to detect for forensic scientists due to rapid metabolism to endogenous levels of GHB. We recently discovered a new major metabolite, 2, of GHB (1 that could potentially extend the analytical detection window for GHB intoxications. Herein we disclose synthetic procedures based on a Koenigs–Knorr glucuronidation approach that provides GHB glucuronide 2 and a deuterium-labelled analogue d4-2 of high purity suitable for analytical chemistry. In addition, we have assessed the stability of GHB glucuronide 2 by mimicking the natural pH range for urine, which is of importance in the development of new analytical methods. Using NMR we show that GHB glucuronide 2 is highly stable towards aqueous hydrolysis within the pH range normally observed for urine even at elevated temperature.

  11. Newly Identified Targets of Aspirin and Its Primary Metabolite, Salicylic Acid.

    Klessig, Daniel F

    2016-04-01

    Salicylic acid (SA) is a plant hormone, which influences several physiological processes, and is a critical modulator of multiple levels of immunity in plants. Several high-throughput screens, which were developed to identify SA-binding proteins through which SA mediates its many physiological effects in plants, uncovered several novel targets of aspirin and its primary metabolite, SA, in humans. These include glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and high mobility group box 1 (HMGB1), two proteins associated with some of the most prevalent and devastating human diseases. In addition, natural and synthetic SA derivatives were discovered, which are much more potent than SA at inhibiting the disease-associated activities of these targets. PMID:26954428

  12. Enhanced L-lactic acid production in Lactobacillus paracasei by exogenous proline addition based on comparative metabolite profiling analysis.

    Tian, Xiwei; Wang, Yonghong; Chu, Ju; Zhuang, Yingping; Zhang, Siliang

    2016-03-01

    This study investigated cell physiological and metabolic responses of Lactobacillus paracasei to osmotic stresses. Both cellular fatty acid composition and metabolite profiling were responded by increasing unsaturated and epoxy-fatty acid proportions, as well as accumulating some specific intracellular metabolites. Simultaneously, metabolite profiling was adopted to rationally and systematically discover potential osmoprotectants. Consequently, exogenous addition of proline or aspartate was validated to be a feasible and efficacious approach to improve cell growth under hyperosmotic stress in shake flasks. Particularly, with 5-L cultivation system, L-lactic acid concentration increased from 108 to 150 g/L during the following 16-h fermentation in 2 g/L proline addition group, while it only increased from 110 to 140 g/L in no proline addition group. Moreover, glucose consumption rate with proline addition reached 3.49 g/L/h during this phase, 35.8 % higher than that with no proline addition. However, extreme high osmotic pressure would significantly limit the osmoprotection of proline, and the osmolality threshold for L. paracasei was approximately 3600 mOsm/kg. It was suggested that proline principally played a role as a compatible solute accumulated in the cell for hyperosmotic preservation. The strategies of exploiting osmotic protectant with metabolite profiling and enhancing L-lactic acid production by osmoprotectant addition would be potential to provide a new insight for other microorganisms and organic acids production. PMID:26658821

  13. Concentrations of the urinary pyrethroid metabolite 3-phenoxybenzoic acid in farm worker families in the MICASA study

    Indoor pesticide exposure is a growing concern, particularly from pyrethroids, a commonly used class of pesticides. Pyrethroid concentrations may be especially high in homes of immigrant farm worker families who often live in close proximity to agricultural fields, and are faced with poor housing conditions, causing higher pest infestation and more pesticide use. We investigate exposure of farm worker families to pyrethroids in a study of mothers and children living in Mendota, CA within the population-based Mexican Immigration to California: Agricultural Safety and Acculturation (MICASA) Study. We present pyrethroid exposure based on an ELISA analysis of urinary metabolite 3-phenoxybenzoic acid (3PBA) levels among 105 women and 103 children. The median urinary 3PBA levels (children=2.56 ug/g creatinine, mothers=1.46 ug/g creatinine) were higher than those reported in population based studies for the United States general population, but similar to or lower than studies with known high levels of pyrethroid exposure. A positive association was evident between poor housing conditions and the urinary metabolite levels, showing that poor housing conditions are a contributing factor to the higher levels of 3PBA seen in the urine of these farm worker families. Further research is warranted to fully investigate sources of exposure. - Highlights: • We investigate exposure of farm worker families to pyrethroids. • We present pyrethroid exposure based on an ELISA analysis of urinary 3PBA levels. • 3PBA levels were higher than those reported for the U.S. general population. • Poor housing conditions may be associated with pyrethroid exposure

  14. Concentrations of the urinary pyrethroid metabolite 3-phenoxybenzoic acid in farm worker families in the MICASA study

    Trunnelle, Kelly J., E-mail: kjtrunnelle@ucdavis.edu [Department of Environmental Toxicology, University of California, Davis 1 Shields Avenue, Davis, CA 95616 (United States); Bennett, Deborah H. [Department of Public Health Sciences, University of California, Davis, CA 95616 (United States); Ahn, Ki Chang [Department of Entomology and Cancer Center, University of California, Davis 1 Shields Avenue, Davis, CA 95616 (United States); Schenker, Marc B. [Department of Public Health Sciences, University of California, Davis, CA 95616 (United States); Tancredi, Daniel J. [Department of Pediatrics, University of California, Davis School of Medicine, 4610 X Street Sacramento, CA 95817 (United States); Gee, Shirley J. [Department of Entomology and Cancer Center, University of California, Davis 1 Shields Avenue, Davis, CA 95616 (United States); Stoecklin-Marois, Maria T. [Department of Public Health Sciences, University of California, Davis, CA 95616 (United States); Hammock, Bruce D. [Department of Entomology and Cancer Center, University of California, Davis 1 Shields Avenue, Davis, CA 95616 (United States)

    2014-05-01

    Indoor pesticide exposure is a growing concern, particularly from pyrethroids, a commonly used class of pesticides. Pyrethroid concentrations may be especially high in homes of immigrant farm worker families who often live in close proximity to agricultural fields, and are faced with poor housing conditions, causing higher pest infestation and more pesticide use. We investigate exposure of farm worker families to pyrethroids in a study of mothers and children living in Mendota, CA within the population-based Mexican Immigration to California: Agricultural Safety and Acculturation (MICASA) Study. We present pyrethroid exposure based on an ELISA analysis of urinary metabolite 3-phenoxybenzoic acid (3PBA) levels among 105 women and 103 children. The median urinary 3PBA levels (children=2.56 ug/g creatinine, mothers=1.46 ug/g creatinine) were higher than those reported in population based studies for the United States general population, but similar to or lower than studies with known high levels of pyrethroid exposure. A positive association was evident between poor housing conditions and the urinary metabolite levels, showing that poor housing conditions are a contributing factor to the higher levels of 3PBA seen in the urine of these farm worker families. Further research is warranted to fully investigate sources of exposure. - Highlights: • We investigate exposure of farm worker families to pyrethroids. • We present pyrethroid exposure based on an ELISA analysis of urinary 3PBA levels. • 3PBA levels were higher than those reported for the U.S. general population. • Poor housing conditions may be associated with pyrethroid exposure.

  15. Glucuronic acid and the ethanol metabolite ethyl-glucuronide cause toll-like receptor 4 activation and enhanced pain.

    Lewis, Susannah S; Hutchinson, Mark R; Zhang, Yingning; Hund, Dana K; Maier, Steven F; Rice, Kenner C; Watkins, Linda R

    2013-05-01

    We have previously observed that the non-opioid morphine metabolite, morphine-3-glucuronide, enhances pain via a toll-like receptor 4 (TLR4) dependent mechanism. The present studies were undertaken to determine whether TLR4-dependent pain enhancement generalizes to other classes of glucuronide metabolites. In silico modeling predicted that glucuronic acid alone and ethyl glucuronide, a minor but long-lasting ethanol metabolite, would dock to the same MD-2 portion of the TLR4 receptor complex previously characterized as the docking site for morphine-3-glucuronide. Glucuronic acid, ethyl glucuronide and ethanol all caused an increase in TLR4-dependent reporter protein expression in a cell line transfected with TLR4 and associated co-signaling molecules. Glucuronic acid-, ethyl glucuronide-, and ethanol-induced increases in TLR4 signaling were blocked by the TLR4 antagonists LPS-RS and (+)-naloxone. Glucuronic acid and ethyl glucuronide both caused allodynia following intrathecal injection in rats, which was blocked by intrathecal co-administration of the TLR4 antagonist LPS-RS. The finding that ethyl glucuronide can cause TLR4-dependent pain could have implications for human conditions such as hangover headache and alcohol withdrawal hyperalgesia, as well as suggesting that other classes of glucuronide metabolites could have similar effects. PMID:23348028

  16. Flavonoid metabolite 3-(3-hydroxyphenyl)propionic acid formed by human microflora decreases arterial blood pressure in rats.

    Najmanová, Iveta; Pourová, Jana; Vopršalová, Marie; Pilařová, Veronika; Semecký, Vladimír; Nováková, Lucie; Mladěnka, Přemysl

    2016-05-01

    There are reports of positive effects of quercetin on cardiovascular pathologies, however, mainly due to its low biovailability, the mechanism remains elusive. Here, we report that one metabolite formed by human microflora (3-(3-hydroxyphenyl)propionic acid)relaxed isolated rat aorta and decreased arterial blood pressure in rats. PMID:26790841

  17. [Effect of phenibut on the content of monoamines, their metabolites, and neurotransmitter amino acids in rat brain structures].

    Borodkina, L E; Kudrin, V S; Klodt, P M; Narkevich, V B; Tiurenkov, I N

    2009-01-01

    Effects of the nootropic drug phenibut, which is a structural analog of gamma-aminobutyric acid (GABA), on the content of monoamines, their metabolites, and neurotransmitter amino acids in brain structures have been studied on Wistar rats. It is established that a single administration of phenibut in a dose of 25 mg/kg (i.p.) produces a statistically significant increase in the content of dopamine metabolite (3,4-dioxyphenylacetic acid) and the retarding amino acid taurine in striatum. At the same time, phenibut did not significantly influence the levels of GABA, serotonin, and dopamine in various brain structures and produce a moderate decrease in the level of norepinephrine in the hippocampus. PMID:19334514

  18. Metabolite changes during natural and lactic acid bacteria fermentations in pastes of soybeans and soybean–maize blends

    Ng'ong'ola-Manani, Tinna A; Østlie, Hilde Marit; Mwangwela, Agnes M.; Wicklund, Trude

    2014-01-01

    The effect of natural and lactic acid bacteria (LAB) fermentation processes on metabolite changes in pastes of soybeans and soybean–maize blends was studied. Pastes composed of 100% soybeans, 90% soybeans and 10% maize, and 75% soybeans and 25% maize were naturally fermented (NFP), and were fermented by lactic acid bacteria (LFP). LAB fermentation processes were facilitated through back-slopping using a traditional fermented gruel, thobwa as an inoculum. Naturally fermented ...

  19. Isolation and Selection of Anti-Candida albicans Metabolites Producing Lactic Acid Bacteria from Various Sources

    Tanes SUNGSRI

    2015-02-01

    Full Text Available Five hundred and fifty-two of lactic acid bacteria (LAB have been isolated and screened from fermented foods, natural sources and dairy effluents on De Mann Rogosa Sharpe (MRS agar. Fifty-one isolates, in the percentile of 9.24, produced the secondary metabolites that could inhibit the growth of Candida albicans BCC6120 by using dual culture overlay assay. The culture broth of LAB, moreover, showed anti-C. albicans activity in acidic condition at pH range of 3.0-5.0 by using agar well diffusion method. Interestingly, the isolate L-47-2 showed much more colonization surrounding the surface of sterile toothpick and test tube when growing in MRS broth. The identification of isolate L-47-2 by morphological and biochemical characteristics using API 50 CHL Test Kit and further confirmed by 16S rRNA gene sequence analysis revealed that isolate L47-2 was similar to Lactobacillus paracasei with 99% nucleotide identity.    

  20. Polycyclic Aromatic Acids Are Primary Metabolites of Alkyl-PAHs-A Case Study with Nereis diversicolor.

    Malmquist, Linus M V; Selck, Henriette; Jørgensen, Kåre B; Christensen, Jan H

    2015-05-01

    Although concentrations of alkylated polycyclic aromatic hydrocarbons (alkyl-PAHs) in oil-contaminated sediments are higher than those of unsubstituted PAHs, only little attention has been given to metabolism and ecotoxicity of alkyl-PAHs. In this study we demonstrated that metabolism of alkyl-PAHs primarily forms polycyclic aromatic acids (PAAs). We generalize this to other alkyl-PAHs, based on literature and the present study of the metabolism of 1-methylphenanthrene, 3,6-dimethylphenanthrene, and 1-, 2-, 3-, and 6-methylchrysene related to their unsubstituted parent PAHs. Also, we observed that body burdens and production of PAAs was related to the position of the methyl group, showing the same isomer specific preferences as for microbial degradation of alkyl-PAHs. We detected a high production of PAAs, and larger metabolism of alkyl-PAHs than their unsubstituted parent PAHs. We therefore propose that carboxylic acid metabolites of alkyl-PAHs have the potential of constituting a new class of contaminants in marine waters that needs attention in relation to ecological risk assessments. PMID:25827176

  1. Formation of 4'-carboxyl acid metabolite of imrecoxib by rat liver microsomes

    Hai-yan XU; Peng ZHANG; Ai-shen GONG; Yu-ming SUN; Feng-ming CHU; Zong-ru GUO; Da-fang ZHONG

    2006-01-01

    Aim:Imrecoxib is a novel and moderately selective COX-2 inhibitor.The aim of the present in vitro investigation was to study the formation of the major metabolite 4'-carboxylic acid imrecoxib (M2) and identify the enzyrne(s) involved in the reaction.Methods:The formation of M2 was studied in rat liver cytosol in the absence or presence of liver microsomes.The formed metabolite was identified and quantified by LC/MSn.In addition,to characterize the cytochrome P450 (CYP) isozymes involved in M2 formation,the effects of typical CYP inhibitors (such as ketoconazle,quinine,α-naphthoflavone, methylpyrazole,and cimetidine) on the formation rate of M2 were investigated.Results:The formation of M2 from 4'hydroxymethyl imrecoxib (M4) was completely dependent on rat liver microsomes and NADPH.Enzyme kinetic studies demonstrated that the formation rate of M2 conformed to monophasic Michaelis-Menten kinetics.Additional experiments showed that the formation of M2 was induced significantly by dexamethasone and lowered by ketoconazole strongly and concentration-dependently.By comparison.other CYP inhibitors.such as α-naphthoflavone,cimetidine,quinine,and methylpyrazole had no inhibitory effects on this metabolic pathway.Conclusion:These biotransformation studies of M4 and imrecoxib in rat liver at the subcellular level showed that the formation of M2 occurs in rat liver microsomes and is NADPH-dependent.The reaction was mainly catalyzed by CYP 3A in untreated rats and in dexamethasone-induced rats.Other CYP,such as CYP 1A,2C,2D,and 2E,seem unlikely to participate in this metabolic pathway.

  2. DNA-strand breaks induced by dimethylarsinic acid, a metabolite of inorganic arsenics, are strongly enhanced by superoxide anion radicals.

    Rin, K; Kawaguchi, K; Yamanaka, K; Tezuka, M; Oku, N; Okada, S

    1995-01-01

    We previously reported that dimethylarsinic acid (DMAA), a major metabolite of inorganic arsenics, induced DNA single-strand breaks (ssb) both in vivo and in cultured alveolar type II (L-132) cells in vitro, possibly via the production of dimethylarsenic peroxyl radicals. Here, the interaction of superoxide anion radicals (O2-) in the induction of ssb in L-132 cells was investigated using paraquat, an O2(-)-producing agent. A significant enhancement of ssb formation was observed in the DMAA-exposed cells when coexposed to paraquat. This enhancement occurred even when post-exposed to DMAA after washing, suggesting that the DMAA exposure caused some modification of DNA such as DNA-adducts, which was recognized by active oxygens to form ssb. An experiment with UV-irradiation, which was likely to induce ssb at the modified region, supported the possibility of DNA modification by DMAA exposure. An ESR study indicated that O2- produced by paraquat in DMAA-exposed cells was more consumed than in non-exposed cells, assumingly through the reaction with the dimethylarsenic-modified region of DNA. The species of active oxygens were estimated by using diethyldithiocarbamate, aminotriazole, diethylmaleate, hydrogen peroxide (H2O2), gamma-irradiation and ethanol. O2- but neither H2O2 nor hydroxyl radicals was very likely to contribute to the ssb-enhancing action of paraquat. PMID:7735248

  3. Is 2-propyl-4-pentenoic acid, a hepatotoxic metabolite of valproate, responsible for valproate-induced hyperammonemia?

    Kondo, T; Ishida, M; Kaneko, S; Hirano, T; Otani, K; Fukushima, Y; Muranaka, H; Koide, N; Yokoyama, M; Nakata, S

    1992-01-01

    To investigate the association between valproate metabolism (VPA) and VPA-induced hyperammonemia together with the contribution of VPA hepatotoxicity risk factors such as young age, polypharmacy, and high serum VPA levels to VPA-induced hyperammonemia, plasma ammonia (NH3) levels, serum levels of VPA and its metabolites, and biochemical parameters were determined in 98 patients treated with VPA (53 monopharmacy cases and 45 polypharmacy cases). In monopharmacy patients, plasma NH3 levels did not depend on age, VPA dosage or serum levels. Serum level of 2-propyl-4-pentenoic acid (4-en) showed a negative correlation with plasma NH3 level in the monopharmacy group. In polypharmacy patients, plasma NH3 levels, serum glutamic pyruvic transaminase, and gamma-glutamyl-transpeptidase were significantly higher, while level/dose VPA ratio, 2-en-VPA serum level, and bilirubin were significantly lower than those in monopharmacy patients. These results suggest that young age and relatively high VPA serum levels within the therapeutic range were unlikely to be risk factors for common hyperammonemia associated with VPA therapy and that 4-en was not causally related to this adverse effect. The decreased serum level of 2-en-VPA in polypharmacy patients may be a reflection of a certain mitochondrial dysfunction, which might be a mechanism of the increased NH3 levels. The changes in biochemical parameters in polypharmacy patients were considered results of the enzyme-inducing activity of coadministered antiepileptic drugs (AEDs). PMID:1350534

  4. Sulphuric acid closure and contribution to nucleation mode particle growth

    M. Boy

    2005-01-01

    Full Text Available Sulphuric acid concentrations were measured and calculated based on pseudo steady state model with corresponding measurements of CO, NOx, O3, SO2, methane and non-methane hydrocarbon (NMHC concentrations as well as solar spectral irradiance and particle number concentrations with size distributions. The measurements were performed as a part of the EU project QUEST (Quantification of Aerosol Nucleation in the European Boundary layer during an intensive field campaign, which was conducted in Hyytiälä, Finland in March–April 2003. In this paper, the closure between measured and calculated H2SO4 concentrations is investigated. Besides that, also the contribution of sulphuric acid to nucleation mode particle growth rates is studied. Hydroxyl and hydroperoxy radical concentrations were determined using a pseudo steady state box model including photo stationary states. The maximum midday OH concentrations ranged between 4.1×105 to 1.8×106 molecules cm-3 and the corresponding values for HO2 were 1.0×107 to 1.5×108 molecules cm-3. The dominant source term for hydroxyl radicals is the reaction of NO with HO2 (56% and the reaction of CO with OH covers around 41% of the sinks. The sulphuric acid source term is the reaction SO2 with OH and the sink term is condensation of sulphuric acid. The closure between measured and calculated sulphuric acid concentrations is achieved with a high agreement to the measured values. In sensitivity studies, we used different values for the non-methane hydrocarbons, the peroxy radicals and nitrogen dioxide. The best fits between calculated and measured values were found by decreasing the NO2 concentration when it exceeded values of 1.5 ppb and doubling the non-methane hydrocarbon concentrations. The ratio, standard deviation and correlation coefficient between measured and calculated sulphuric acid concentrations are 0.99, 0.412 and 0.645, respectively. The maximum midday sulphuric acid concentrations varied between

  5. In vivo relevant mixed urolithins and ellagic acid inhibit phenotypic and molecular colon cancer stem cell features: A new potentiality for ellagitannin metabolites against cancer.

    Núñez-Sánchez, María Ángeles; Karmokar, Ankur; González-Sarrías, Antonio; García-Villalba, Rocío; Tomás-Barberán, Francisco A; García-Conesa, María Teresa; Brown, Karen; Espín, Juan Carlos

    2016-06-01

    Colon cancer stem cells (CSCs) offer a novel paradigm for colorectal cancer (CRC) treatment and dietary polyphenols may contribute to battle these cells. Specifically, polyphenol-derived colon metabolites have the potential to interact with and affect colon CSCs. We herein report the effects against colon CSCs of two mixtures of ellagitannin (ET) metabolites, ellagic acid (EA) and the gut microbiota-derived urolithins (Uro) at concentrations detected in the human colon tissues following the intake of ET-containing products (pomegranate, walnuts). These mixtures reduce phenotypic and molecular features in two models of colon CSCs: Caco-2 cells and primary tumour cells from a patient with CRC. The mixture containing mostly Uro-A (85% Uro-A, 10% Uro-C, 5% EA) was most effective at inhibiting the number and size of colonospheres and aldehyde dehydrogenase activity (ALDH, a marker of chemoresistance) whereas the mixture containing less Uro-A but IsoUro-A and Uro-B (30% Uro-A, 50% IsoUro-A, 10% Uro-B, 5% Uro-C, 5% EA) had some effects on the number and size of colonospheres but not on ALDH. These data support a role for polyphenols metabolites in the control of colon cancer chemoresistance and relapse and encourage the research on the effects of polyphenols against CSCs. PMID:26995228

  6. A novel function for kojic acid, a secondary metabolite from Aspergillus fungi, as antileishmanial agent.

    Ana Paula D Rodrigues

    Full Text Available Kojic acid (KA is a fungal metabolite used as a topical treatment skin-whitening cosmetic agent for melasma in humans; however its potential as an anti-leishmanial agent is unknown. Chemotherapy is one of the most effective treatments for Leishmaniasis. However, the drugs available are expensive, invasive, require long-term treatment and have severe side effects. Thus, the development of new effective leishmanicidal agents is a necessity. In this study we investigated the anti-leishmanial effect of KA on L. amazonensis, following in vitro and in vivo infections. KA (50 μg/mL was found to decrease the growth by 62% (IC50 34 μg/mL and 79% (IC50 27.84 μg/mL of promastigotes and amastigotes in vitro, respectively. Ultrastructural analysis of KA-treated amastigotes showed the presence of vesicles bodies into the flagellar pocket, and an intense intracellular vacuolization and swelling of the mitochondrion. During the in vitro interaction of parasites and the host cell, KA reverses the superoxide anions (O2- inhibitory mechanism promoted by parasite. In addition, 4 weeks after KA-topical formulation treatment of infected animals, a healing process was observed with a high production of collagen fibers and a decrease in parasite burden. Thus, these results demonstrated the great potential of KA as an anti-leishmanial compound.

  7. The effects of anaerobic training in serum lipids and arachidonic acid metabolites

    GEORGIOS KIPREOS

    2010-01-01

    Full Text Available Coronary arteries are subjected daily in high shear stress and manifest atherosclerosis very early in life in comparison to other arteries in the human body. Some factors that are implicated in the evolution and progress of this process are the concentration of lipids and arachidonic acid metabolites, such prostacyclin and thromboxane. It has been reported that those who participate in aerobic activities such as walking, cycling, jogging or brisk walking might have normal values of the mentioned chemical substances. On the other hand, it is reported that the effects of anaerobic and strength activities has negative effects on the vascular endothelium, which is essential for the maintenance of hemostatic balance and the local regulation of vascular tone.Therefore, even although extensive research has been conducted in this field, there are crucial gaps in our knowledge. Consequently, the purpose of this brief review is to describe what is known about the effects of anaerobic activities in which the competitive athletes have participated on the following blood parameters: Total cholesterol, triglycerides, high density lipoprotein cholesterol (HDL - C, low density lipoproteins cholesterol (LDL - C, prostacyclin & thromboxane.

  8. Aspirin's Active Metabolite Salicylic Acid Targets High Mobility Group Box 1 to Modulate Inflammatory Responses.

    Choi, Hyong Woo; Tian, Miaoying; Song, Fei; Venereau, Emilie; Preti, Alessandro; Park, Sang-Wook; Hamilton, Keith; Swapna, G V T; Manohar, Murli; Moreau, Magali; Agresti, Alessandra; Gorzanelli, Andrea; De Marchis, Francesco; Wang, Huang; Antonyak, Marc; Micikas, Robert J; Gentile, Daniel R; Cerione, Richard A; Schroeder, Frank C; Montelione, Gaetano T; Bianchi, Marco E; Klessig, Daniel F

    2015-01-01

    Salicylic acid (SA) and its derivatives have been used for millennia to reduce pain, fever and inflammation. In addition, prophylactic use of acetylsalicylic acid, commonly known as aspirin, reduces the risk of heart attack, stroke and certain cancers. Because aspirin is rapidly de-acetylated by esterases in human plasma, much of aspirin's bioactivity can be attributed to its primary metabolite, SA. Here we demonstrate that human high mobility group box 1 (HMGB1) is a novel SA-binding protein. SA-binding sites on HMGB1 were identified in the HMG-box domains by nuclear magnetic resonance (NMR) spectroscopic studies and confirmed by mutational analysis. Extracellular HMGB1 is a damage-associated molecular pattern molecule (DAMP), with multiple redox states. SA suppresses both the chemoattractant activity of fully reduced HMGB1 and the increased expression of proinflammatory cytokine genes and cyclooxygenase 2 (COX-2) induced by disulfide HMGB1. Natural and synthetic SA derivatives with greater potency for inhibition of HMGB1 were identified, providing proof-of-concept that new molecules with high efficacy against sterile inflammation are attainable. An HMGB1 protein mutated in one of the SA-binding sites identified by NMR chemical shift perturbation studies retained chemoattractant activity, but lost binding of and inhibition by SA and its derivatives, thereby firmly establishing that SA binding to HMGB1 directly suppresses its proinflammatory activities. Identification of HMGB1 as a pharmacological target of SA/aspirin provides new insights into the mechanisms of action of one of the world's longest and most used natural and synthetic drugs. It may also provide an explanation for the protective effects of low-dose aspirin usage. PMID:26101955

  9. Oxygen metabolites stimulate release of high-molecular-weight glycoconjugates by cell and organ cultures of rodent respiratory epithelium via an arachidonic acid-dependent mechanism.

    Adler, K B; Holden-Stauffer, W J; Repine, J E

    1990-01-01

    Several common pulmonary disorders characterized by mucus hypersecretion and airway obstruction may relate to increased levels of inhaled or endogenously generated oxidants (O2 metabolites) in the respiratory tract. We found that O2 metabolites stimulated release of high-molecular-weight glycoconjugates (HMG) by respiratory epithelial cells in vitro through a mechanism involving cyclooxygenase metabolism of arachidonic acid. Noncytolytic concentrations of chemically generated O2 metabolites (...

  10. The Oxidized Linoleic Acid Metabolite-Cytochrome P450 System is Active in Biopsies from Patients with Inflammatory Dental Pain

    Ruparel, Shivani; Hargreaves, Kenneth. M.; Eskander, Michael; Rowan, Spencer; Almeida, Jose F. A.; Roman, Linda; Henry, Michael A.

    2013-01-01

    Endogenous TRPV1 agonists such as oxidized linoleic acid metabolites (OLAMs) and the enzymes releasing them [e.g., cytochrome P450 (CYP)], are up-regulated following inflammation in the rat. However, it is not known if such agonists are elevated in human inflammatory pain conditions. Since TRPV1 is expressed in human dental pulp nociceptors, we hypothesized that OLAM-CYP machinery is active in this tissue type and is increased under painful inflammatory conditions such as irreversible pulpiti...

  11. Sulphuric acid closure and contribution to nucleation mode particle growth

    M. Boy

    2004-10-01

    Full Text Available Sulphuric acid concentrations were measured and calculated based on pseudo steady state model with corresponding measurements of CO, NOx, O3, SO2, methane and non-methane hydrocarbon (NMHC concentrations as well as solar spectral irradiance and particle number concentrations with size distributions. The measurements were performed as a part of the EU project QUEST (Quantification of Aerosol Nucleation in the European Boundary layer during an intensive field campaign, which was conducted in Hyytiälä, Finland in March–April 2003. In this paper, the closure between measured and calculated H2SO4 concentrations is investigated. Besides that, also the contribution of sulphuric acid to nucleation mode particle growth rates is studied. Hydroxyl and hydroperoxy radical concentrations were determined using a pseudo steady state box model including photo stationary states. The maximum midday OH concentrations ranged between 4.1×105 to 1.8×106molecules cm−3 and the corresponding values for HO2 were 1.0×107 to 1.5×108molecules cm−3. The dominant source term for hydroxyl radicals is the reaction of NO with HO2 (56% and the reaction of CO with OH covers around 41% of the sinks. The sulphuric acid source term is the reaction SO2 with OH and the sink term is condensation of sulphuric acid. The closure between measured and calculated sulphuric acid concentrations is achieved with a high agreement to the measured values. In sensitivity studies, we used different values for the non-methane hydrocarbons, the peroxy radicals and nitrogen dioxide. The best fits between calculated and measured values were found by decreasing the NO2 concentration when it exceeded values of 1.5 ppb and doubling the non-methane hydrocarbon concentrations. The ratio, standard deviation and correlation coefficient

  12. Human GAPDH Is a Target of Aspirin's Primary Metabolite Salicylic Acid and Its Derivatives.

    Hyong Woo Choi

    Full Text Available The plant hormone salicylic acid (SA controls several physiological processes and is a key regulator of multiple levels of plant immunity. To decipher the mechanisms through which SA's multiple physiological effects are mediated, particularly in immunity, two high-throughput screens were developed to identify SA-binding proteins (SABPs. Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDH from plants (Arabidopsis thaliana was identified in these screens. Similar screens and subsequent analyses using SA analogs, in conjunction with either a photoaffinity labeling technique or surface plasmon resonance-based technology, established that human GAPDH (HsGAPDH also binds SA. In addition to its central role in glycolysis, HsGAPDH participates in several pathological processes, including viral replication and neuronal cell death. The anti-Parkinson's drug deprenyl has been shown to suppress nuclear translocation of HsGAPDH, an early step in cell death and the resulting cell death induced by the DNA alkylating agent N-methyl-N'-nitro-N-nitrosoguanidine. Here, we demonstrate that SA, which is the primary metabolite of aspirin (acetyl SA and is likely responsible for many of its pharmacological effects, also suppresses nuclear translocation of HsGAPDH and cell death. Analysis of two synthetic SA derivatives and two classes of compounds from the Chinese medicinal herb Glycyrrhiza foetida (licorice, glycyrrhizin and the SA-derivatives amorfrutins, revealed that they not only appear to bind HsGAPDH more tightly than SA, but also exhibit a greater ability to suppress translocation of HsGAPDH to the nucleus and cell death.

  13. Human GAPDH Is a Target of Aspirin's Primary Metabolite Salicylic Acid and Its Derivatives.

    Choi, Hyong Woo; Tian, Miaoying; Manohar, Murli; Harraz, Maged M; Park, Sang-Wook; Schroeder, Frank C; Snyder, Solomon H; Klessig, Daniel F

    2015-01-01

    The plant hormone salicylic acid (SA) controls several physiological processes and is a key regulator of multiple levels of plant immunity. To decipher the mechanisms through which SA's multiple physiological effects are mediated, particularly in immunity, two high-throughput screens were developed to identify SA-binding proteins (SABPs). Glyceraldehyde 3-Phosphate Dehydrogenase (GAPDH) from plants (Arabidopsis thaliana) was identified in these screens. Similar screens and subsequent analyses using SA analogs, in conjunction with either a photoaffinity labeling technique or surface plasmon resonance-based technology, established that human GAPDH (HsGAPDH) also binds SA. In addition to its central role in glycolysis, HsGAPDH participates in several pathological processes, including viral replication and neuronal cell death. The anti-Parkinson's drug deprenyl has been shown to suppress nuclear translocation of HsGAPDH, an early step in cell death and the resulting cell death induced by the DNA alkylating agent N-methyl-N'-nitro-N-nitrosoguanidine. Here, we demonstrate that SA, which is the primary metabolite of aspirin (acetyl SA) and is likely responsible for many of its pharmacological effects, also suppresses nuclear translocation of HsGAPDH and cell death. Analysis of two synthetic SA derivatives and two classes of compounds from the Chinese medicinal herb Glycyrrhiza foetida (licorice), glycyrrhizin and the SA-derivatives amorfrutins, revealed that they not only appear to bind HsGAPDH more tightly than SA, but also exhibit a greater ability to suppress translocation of HsGAPDH to the nucleus and cell death. PMID:26606248

  14. Characterization of metabolites in rats after intravenous administration of salvianolic acid for injection by ultra-performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry.

    Miao, Jingzhuo; Sun, Wanyang; Huang, Jingyi; Liu, Xiaolin; Li, Shuming; Han, Xiaoping; Tong, Ling; Sun, Guoxiang

    2016-09-01

    It is an essential requirement to clarify the metabolites of traditional Chinese medicine (TCM) injections, which contain numerous ingredients, to assess their safe and effective use in clinic. Salvianolic acid for injection (SAFI), made from hydrophilic phenolic acids in Salvia miltiorrhiza Bunge, has been widely used for the treatment of cerebrovascular diseases, but information on its metabolites in vivo is still lacking. In the present study, we aimed to holistically characterize the metabolites of the main active ingredients in rat plasma, bile, urine and feces following intravenous administration of SAFI. An ultra-performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry (UPLC/Q-TOF-MS) method was developed. Combining information on retention behaviors, multistage mass spectra and literature data, a total of eight prototypes and 52 metabolites were tentatively characterized. Metabolites originated from rosmarinic acid and salvianolic acid B comprised the majority of identified compounds. Meanwhile, four metabolites derived from salvianolic acid D and five from salvianolic acid B are reported for the first time. This study revealed that methylation, sulfation and glucuronidation were the major metabolic pathways of phenolic acids in SAFI in vivo. Furthermore, the developed UPLC/Q-TOF-MS method could also benefit the metabolic investigation of extracts and preparations in TCM with hydrophilic ingredients. Copyright © 2016 John Wiley & Sons, Ltd. PMID:26910272

  15. Effect of ascorbic acid supplementation on nitric oxide metabolites and systolic blood pressure in rats exposed to lead

    Mohammad Amani

    2010-01-01

    Full Text Available Background: Extended exposure to low levels of lead causes high blood pressure in human and laboratory animals. The mechanism is not completely recognized, but it is relatively implicated with generation of free radicals, oxidant agents such as ROS, and decrease of available nitric oxide (NO. In this study, we have demonstrated the effect of ascorbic acid as an antioxidant on nitric oxide metabolites and systolic blood pressure in rats exposed to low levels of lead. Materials and Methods: The adult male Wistar rats weighing 200-250 g were divided into four groups: control, lead acetate (receiving 100 ppm lead acetate in drinking water, lead acetate plus ascorbic acid (receiving 100 ppm lead acetate and 1 g/l ascorbic acid in drinking water, and ascorbic acid (receiving 1 g/l ascorbic acid in drinking water groups. The animals were anesthetized with ketamin/xylazine (50 and 7 mg/kg, respectively, ip and systolic blood pressure was then measured from the tail of the animals by a sphygmomanometer. Nitric oxide levels in serum were measured indirectly by evaluation of its stable metabolites (total nitrite and nitrate (NOc. Results: After 8 and 12 weeks, systolic blood pressure in the lead acetate group was significantly elevated compared to the control group. Ascorbic acid supplementation could prevent the systolic blood pressure rise in the lead acetate plus ascorbic acid group and there was no significant difference relative to the control group. The serum NOc levels in lead acetate group significantly decreased in relation to the control group, but this reduction was not significantly different between the lead acetate plus ascorbic acid group and the control group. Conclusion: Results of this study suggest that ascorbic acid as an antioxidant prevents the lead induced hypertension. This effect may be mediated by inhibition of NOc oxidation and thereby increasing availability of NO.

  16. Enhancement of Urinary Elimination of 3-Bromobenzanthrone Metabolites by Oral Supplementation of Ascorbic Acid in Guinea Pigs

    RAVINDRA P.SINGH; RAJ KHANNA; SUBHASH K. KHANNA; MUKUL DAS

    2004-01-01

    3-Bromobenzanthrone (3-BBA), an anthraquinone intermediate dye, is extensively used in textile industry. Since, our prior studies have shown that 3-BBA caused significant depletion of ascorbic acid (AsA) levels, the effect of exogenous supplementation of AsA on the urinary elimination of 3-BBA metabolites was investigated.Method Guinea pigs were treated with single oral dose of 3-BBA (50 mg/kg b. Wt.) in groundnut oil while another group was treated with single oral dose of 3-BBA (50 mg/kg b. Wt.) along with 3 day prior and post oral supplementation of AsA. Control groups were either treated with groundnut oil or AsA alone. Urine from individual animals was collected, extracted and analysed on HPTLC.Results The highest elimination of 3-BBA (75 (g) was found to be in 0-24 h urine fraction which decreased to 18 (g and 5 (g in the two subsequent 24 hourly fractions of urine. Exogenous supplementation of AsA increased the total urinary elimination of 3-BBA by almost 77%. A total of 10 fluorescent metabolites excluding the parent compound were eliminated in the urine of guinea pigs treated with 3-BBA. Densitometric scanning of chromatogram showed different peaks at Rf 0.18, 0.22, 0.27, 0.34, 0.40, 0.48, 0.56, 0.66, 0.72, 0.80, and 0.95 which were eliminated and marked as urinary metabolite 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, and 11 respectively. AsA not only significantly enhanced the elimination of 3-BBA metabolites but also modified the pattern of metabolites drastically in 0-6 h, 6-24 h and 24-48 h urine fractions.Conclusion These results indicate that AsA may be useful in protecting the toxicity of 3-BBA by fascilitating the urinary metabolite(s) excretion of 3-BBA.

  17. Novel nonadride, heptadride and maleic acid metabolites from the byssochlamic acid producer Byssochlamys fulva IMI 40021 - an insight into the biosynthesis of maleidrides.

    Szwalbe, Agnieszka J; Williams, Katherine; O'Flynn, Daniel E; Bailey, Andrew M; Mulholland, Nicholas P; Vincent, Jason L; Willis, Christine L; Cox, Russell J; Simpson, Thomas J

    2015-12-14

    The filamentous fungus Byssochlamys fulva strain IMI 40021 produces (+)-byssochlamic acid 1, its novel dihydroanalogue 2 and four related secondary metabolites. Agnestadrides A, 17 and B, 18 constitute a novel class of seven-membered ring, maleic anhydride-containing (hence termed heptadride) natural products. The putative maleic anhydride precursor 5 for both nonadride and heptadride biosynthesis was isolated as a fermentation product for the first time and its structure confirmed by synthesis. Acid 5 undergoes facile decarboxylation to anhydride 6. The generic term maleidrides is proposed to encompass biosynthetically-related compounds containing maleic anhydride moieties fused to an alicyclic ring, varying in size and substituents. PMID:26452099

  18. The Influence of Clay on the Rate of Decay of Amino Acid Metabolites Synthesized in Soils during Decomposition of Cellulose

    Sørensen, Lasse Holst

    1975-01-01

    amounts of labelled amino acid C in the soils were proportional to their content of silt + clay. After 30 days of incubation labelled amino acid C remaining in the soil with the lowest content of silt + clay constituted 6 per cent of the carbon added in cellulose, as compared with 18 per cent in the soil...... increase in the CO2 evolution caused by the treatments in the different soils was, however, not related to the amount of silt + clay, and a high content of this material did not protect organic material against the effect of the treatments. is concluded that the silt + clay fraction ensures stabilization......14C-labelled cellulose was added to seven different soils containing silt + clay (particles <0.02 mm) in amounts which varied from 8 to 75 per cent. The cellulose was allowed to decompose, and the amounts of labelled C transformed into metabolites hydrolyzable into amino acids were determined. The...

  19. Metabolomics reveals amino acids contribute to variation in response to simvastatin treatment.

    Miles Trupp

    Full Text Available UNLABELLED: Statins are widely prescribed for reducing LDL-cholesterol (C and risk for cardiovascular disease (CVD, but there is considerable variation in therapeutic response. We used a gas chromatography-time-of-flight mass-spectrometry-based metabolomics platform to evaluate global effects of simvastatin on intermediary metabolism. Analyses were conducted in 148 participants in the Cholesterol and Pharmacogenetics study who were profiled pre and six weeks post treatment with 40 mg/day simvastatin: 100 randomly selected from the full range of the LDL-C response distribution and 24 each from the top and bottom 10% of this distribution ("good" and "poor" responders, respectively. The metabolic signature of drug exposure in the full range of responders included essential amino acids, lauric acid (p<0.0055, q<0.055, and alpha-tocopherol (p<0.0003, q<0.017. Using the HumanCyc database and pathway enrichment analysis, we observed that the metabolites of drug exposure were enriched for the pathway class amino acid degradation (p<0.0032. Metabolites whose change correlated with LDL-C lowering response to simvastatin in the full range responders included cystine, urea cycle intermediates, and the dibasic amino acids ornithine, citrulline and lysine. These dibasic amino acids share plasma membrane transporters with arginine, the rate-limiting substrate for nitric oxide synthase (NOS, a critical mediator of cardiovascular health. Baseline metabolic profiles of the good and poor responders were analyzed by orthogonal partial least square discriminant analysis so as to determine the metabolites that best separated the two response groups and could be predictive of LDL-C response. Among these were xanthine, 2-hydroxyvaleric acid, succinic acid, stearic acid, and fructose. Together, the findings from this study indicate that clusters of metabolites involved in multiple pathways not directly connected with cholesterol metabolism may play a role in modulating

  20. Novel β-cyclodextrin modified CdTe quantum dots as fluorescence nanosensor for acetylsalicylic acid and metabolites

    β-Cyclodextrin was modified with 11-[(ethoxycarbonyl)thio]undecanoic acid and used as a capping agent, together with mercaptosuccinic acid, to prepare water-stable CdTe quantum dots. The water soluble quantum dot obtained displays fluorescence with a maximum emission at 425 nm (under excitation at 300 nm) with lifetimes of 0.53, 4.8, 181, and 44.1 ns, respectively. The S-βCD-MSA-CdTe can act as a nanoprobe that is due to the affinity of the cyclodextrin moiety for selected substances such as acetylsalicylic acid (ASA) and its metabolites as foreign species. The fluorescence of the S-βCD-MSA-CdTe is enhanced on addition of ASA. Linear calibration plots are observed with ASA in concentrations between 0 and 1 mg/l, with a limit of detection at 8.5 × 10−9 mol/l (1.5 ng/ml) and a precision as relative standard deviation of 1% (0.05 mg/l). The interference effect of certain compounds as ascorbic acid and its main metabolites such as salicylic, gentisic and salicyluric acid upon the obtained procedure was studied. Highlights: ► Nanosensors constituted by CdTe quantum dots capped with modified cyclodextrin. ► This nanomaterial shows fluorescence properties compatible with a semiconductor quantum dot. ► The nanosensor shows fluorescence enhancement when inclusion complexes are formed with acetylsalicylic acid. ► This nanomaterial has nanosensor potential taking into consideration the formation stability of the inclusion complex.

  1. Novel {beta}-cyclodextrin modified CdTe quantum dots as fluorescence nanosensor for acetylsalicylic acid and metabolites

    Algarra, M. [Centro de Geologia do Porto, Faculdade de Ciencias, Universidade do Porto, Rua do Campo Alegre 687, 4169-007 Porto (Portugal); Campos, B.B.; Aguiar, F.R.; Rodriguez-Borges, J.E. [Centro de Investigacao em Quimica (CIQ-UP), Faculdade de Ciencias da Universidade do Porto, Rua do Campo Alegre 687, 169-007 Porto (Portugal); Esteves da Silva, J.C.G., E-mail: jcsilva@fc.up.pt [Centro de Investigacao em Quimica (CIQ-UP), Faculdade de Ciencias da Universidade do Porto, Rua do Campo Alegre 687, 169-007 Porto (Portugal)

    2012-05-01

    {beta}-Cyclodextrin was modified with 11-[(ethoxycarbonyl)thio]undecanoic acid and used as a capping agent, together with mercaptosuccinic acid, to prepare water-stable CdTe quantum dots. The water soluble quantum dot obtained displays fluorescence with a maximum emission at 425 nm (under excitation at 300 nm) with lifetimes of 0.53, 4.8, 181, and 44.1 ns, respectively. The S-{beta}CD-MSA-CdTe can act as a nanoprobe that is due to the affinity of the cyclodextrin moiety for selected substances such as acetylsalicylic acid (ASA) and its metabolites as foreign species. The fluorescence of the S-{beta}CD-MSA-CdTe is enhanced on addition of ASA. Linear calibration plots are observed with ASA in concentrations between 0 and 1 mg/l, with a limit of detection at 8.5 Multiplication-Sign 10{sup -9} mol/l (1.5 ng/ml) and a precision as relative standard deviation of 1% (0.05 mg/l). The interference effect of certain compounds as ascorbic acid and its main metabolites such as salicylic, gentisic and salicyluric acid upon the obtained procedure was studied. Highlights: Black-Right-Pointing-Pointer Nanosensors constituted by CdTe quantum dots capped with modified cyclodextrin. Black-Right-Pointing-Pointer This nanomaterial shows fluorescence properties compatible with a semiconductor quantum dot. Black-Right-Pointing-Pointer The nanosensor shows fluorescence enhancement when inclusion complexes are formed with acetylsalicylic acid. Black-Right-Pointing-Pointer This nanomaterial has nanosensor potential taking into consideration the formation stability of the inclusion complex.

  2. Human urine certified reference material CZ 6010: creatinine and toluene metabolites (hippuric acid and o-cresol) and a benzene metabolite (phenol)

    Šperlingová, I.; Dabrowská, L.; Stránský, V.; Kučera, Jan; Tichý, M.

    2007-01-01

    Roč. 387, č. 7 (2007), s. 2419-2424. ISSN 1618-2642 Institutional research plan: CEZ:AV0Z10480505 Keywords : toluene metabolites * benzene metabolite * human urine Subject RIV: BG - Nuclear, Atomic and Molecular Physics, Colliders Impact factor: 2.867, year: 2007

  3. From microbe to man: the role of microbial short chain fatty acid metabolites in host cell biology.

    Natarajan, Niranjana; Pluznick, Jennifer L

    2014-12-01

    Recent studies have highlighted a myriad of ways in which the activity and composition of the gut microbiota can affect the host organism. A primary way in which the gut microbiota affect host physiology is by the production of metabolites, such as short-chain fatty acids (SCFAs), which are subsequently absorbed into the bloodstream of the host. Although recent studies have begun to unravel the ways in which gut microbial SCFAs affect host physiology, less is understood regarding the underlying cell biological mechanisms. In this review, we will outline the known receptors and transporters for SCFAs, and review what is known about the cell biological effects of microbial SCFAs. PMID:25273884

  4. Preliminary study to prepare a reference material of styrene metabolites – mandelic acid and phenolglyoxilic acid – in human urine

    Šperlingová, I.; Dabrowská, L.; Stránský, V.; Kučera, Jan; Tichý, M.

    2003-01-01

    Roč. 8, 3-4 (2003), s. 113-116. ISSN 0949-1775 Institutional research plan: CEZ:AV0Z1048901 Keywords : reference material * human urine * styrene metabolites Subject RIV: BG - Nuclear, Atomic and Molecular Physics, Colliders Impact factor: 0.637, year: 2003

  5. Effects of intermediate metabolite carboxylic acids of TCA cycle on Microcystis with overproduction of phycocyanin.

    Bai, Shijie; Dai, Jingcheng; Xia, Ming; Ruan, Jing; Wei, Hehong; Yu, Dianzhen; Li, Ronghui; Jing, Hongmei; Tian, Chunyuan; Song, Lirong; Qiu, Dongru

    2015-04-01

    Toxic Microcystis species are the main bloom-forming cyanobacteria in freshwaters. It is imperative to develop efficient techniques to control these notorious harmful algal blooms (HABs). Here, we present a simple, efficient, and environmentally safe algicidal way to control Microcystis blooms, by using intermediate carboxylic acids from the tricarboxylic acid (TCA) cycle. The citric acid, alpha-ketoglutaric acid, succinic acid, fumaric acid, and malic acid all exhibited strong algicidal effects, and particularly succinic acid could cause the rapid lysis of Microcystis in a few hours. It is revealed that the Microcystis-lysing activity of succinic acid and other carboxylic acids was due to their strong acidic activity. Interestingly, the acid-lysed Microcystis cells released large amounts of phycocyanin, about 27-fold higher than those of the control. On the other hand, the transcription of mcyA and mcyD of the microcystin biosynthesis operon was not upregulated by addition of alpha-ketoglutaric acid and other carboxylic acids. Consider the environmental safety of intermediate carboxylic acids. We propose that administration of TCA cycle organic acids may not only provide an algicidal method with high efficiency and environmental safety but also serve as an applicable way to produce and extract phycocyanin from cyanobacterial biomass. PMID:25342454

  6. The Oxidized Linoleic Acid Metabolite-Cytochrome P450 System is Active in Biopsies from Patients with Inflammatory Dental Pain

    Ruparel, Shivani; Hargreaves, Kenneth M.; Eskander, Michael; Rowan, Spencer; de Almeida, Jose F.A.; Roman, Linda; Henry, Michael A.

    2013-01-01

    Endogenous TRPV1 agonists such as oxidized linoleic acid metabolites (OLAMs) and the enzymes releasing them [e.g., cytochrome P450 (CYP)], are up-regulated following inflammation in the rat. However, it is not known if such agonists are elevated in human inflammatory pain conditions. Since TRPV1 is expressed in human dental pulp nociceptors, we hypothesized that OLAM-CYP machinery is active in this tissue type and is increased under painful inflammatory conditions such as irreversible pulpitis (IP). The aim of this study was to compare CYP expression and linoleic acid (LA) metabolism in normal versus inflamed human dental pulp. Our data showed that exogenous LA metabolism was significantly increased in IP tissues compared to normal tissues and that pretreatment with a CYP inhibitor, ketoconazole, significantly inhibited LA metabolism. Additionally, extracts obtained from LA-treated inflamed tissues, evoked significant inward currents in TG neurons, and were blocked by pretreatment with the TRPV1 antagonist, IRTX. Moreover, extracts obtained from ketoconazole-pretreated inflamed tissues significantly reduced inward currents in TG neurons. These data suggest that LA metabolites produced in human inflamed tissues act as TRPV1 agonists and that the metabolite production can be targeted by CYP inhibition. In addition, immunohistochemical analysis of two CYP isoforms, CYP2J and CYP3A1, were shown to be predominately expressed in immune cells infiltrating the inflamed dental pulp, emphasizing the paracrine role of CYP enzymes in OLAM regulation. Collectively, our data indicates that the machinery responsible for OLAM production is up-regulated during inflammation and can be targeted to develop potential analgesics for inflammatory-induced dental pain. PMID:23867730

  7. A sensitive GC-EIMS method for simultaneous detection and quantification of JWH-018 and JWH-073 carboxylic acid and hydroxy metabolites in urine.

    Paul, Buddha D; Bosy, Thomas

    2015-04-01

    Synthetic cannabinoids, including JWH-018 and JWH-073, belong to a class of aminoalkylindoles (AAIs) that are smoked to produce an effect similar to tetrahydrocannabinol. Compounds in this class are often collectively known as 'Spice'. After ingestion, these compounds are extensively metabolized to their hydroxy and carboxylic acid metabolites. During forensic analysis, detection of these metabolites in urine is an indication of past exposure to the parent compounds. The analytical process involved hydrolysis of conjugated metabolites by glucuronidase, solvent extraction, derivatization by trifluoroacetic anhydride and hexafluoroisopropanol and GC-EIMS detection. Identification of the unknown was based on the criteria of GC retention time within ±2% and mass spectral ion ratio within ±20% of that of a standard. Deuterated internal standards of the carboxylic acid metabolites were used for quantification. The acid (JWH-018-COOH, JWH-073-COOH) and hydroxy (JWH-018-OH, JWH-073-OH) metabolites were linear over the concentration range of 0.1-10 and 0.2-10 ng/mL, respectively, with a correlation coefficient-square, R(2) > 0.999 (N = 5). Extraction recoveries of the metabolites were within 79 and 87%. The method was applied to 17 urine specimens collected as part of a military law enforcement investigation. Nine of the specimens tested positive for one or more of the metabolites. When the procedure was extended to screen other AAI compounds, two of the specimens were found to contain JWH-210, JWH-250 (JWH-302 or JWH-201) and JWH-250 (C4 isomers). The GC-EIMS method presented here was found to be suitable for detecting JWH-018 and JWH-073 metabolites and other AAI compounds in urine. PMID:25691387

  8. Deuterated biogenic amine metabolites: preparation of ring-deuterated 4-hydroxy-3-methoxymandelic acid

    Selected ion monitoring (SIM) assays are now widely used for measuring the concentrations of monoamine neurotransmitters and their metabolites in urine, blood, cerebrospinal fluid, and brain extracts. Although these assays have a high degree of sensitivity and specificity, the successful application of them to routine work requires the availability of suitable internal standards. A one step process, using the platinum catalyzed reaction between aromatic ring protons and deuterium oxide is described to prepare, in reasonable yield, crystalline ring-deuterated VMA suitable for use as an internal standard in SIM assays. (author)

  9. Catabolism of Branched Chain Amino Acids Contributes Significantly to Synthesis of Odd-Chain and Even-Chain Fatty Acids in 3T3-L1 Adipocytes.

    Scott B Crown

    Full Text Available The branched chain amino acids (BCAA valine, leucine and isoleucine have been implicated in a number of diseases including obesity, insulin resistance, and type 2 diabetes mellitus, although the mechanisms are still poorly understood. Adipose tissue plays an important role in BCAA homeostasis by actively metabolizing circulating BCAA. In this work, we have investigated the link between BCAA catabolism and fatty acid synthesis in 3T3-L1 adipocytes using parallel 13C-labeling experiments, mass spectrometry and model-based isotopomer data analysis. Specifically, we performed parallel labeling experiments with four fully 13C-labeled tracers, [U-13C]valine, [U-13C]leucine, [U-13C]isoleucine and [U-13C]glutamine. We measured mass isotopomer distributions of fatty acids and intracellular metabolites by GC-MS and analyzed the data using the isotopomer spectral analysis (ISA framework. We demonstrate that 3T3-L1 adipocytes accumulate significant amounts of even chain length (C14:0, C16:0 and C18:0 and odd chain length (C15:0 and C17:0 fatty acids under standard cell culture conditions. Using a novel GC-MS method, we demonstrate that propionyl-CoA acts as the primer on fatty acid synthase for the production of odd chain fatty acids. BCAA contributed significantly to the production of all fatty acids. Leucine and isoleucine contributed at least 25% to lipogenic acetyl-CoA pool, and valine and isoleucine contributed 100% to lipogenic propionyl-CoA pool. Our results further suggest that low activity of methylmalonyl-CoA mutase and mass action kinetics of propionyl-CoA on fatty acid synthase result in high rates of odd chain fatty acid synthesis in 3T3-L1 cells. Overall, this work provides important new insights into the connection between BCAA catabolism and fatty acid synthesis in adipocytes and underscores the high capacity of adipocytes for metabolizing BCAA.

  10. Generation of novel metabolites of dietary linoleic acid (18:2n6) by guinea pig epidermis

    Although the authors have demonstrated the inability of rat and guinea pig (GP) skin enzyme preparations to desaturate 18:2n6 into gammalinolenic acid (18:3n6) using an in vitro microsomal system, the fate of this dietary essential fatty acid in the GP epidermis is unknown. To explore the fate of 18:2n6, intact tissue slices from GP epidermis were incubated with [1-14C]18:2n6. After incubation, the extracted lipids were transesterified using methanolic-HCL. The fatty acid methyl esters were analyzed using a combination of (i) argentation TLC, scanned using a proportional TLC radioscanner, and (ii) reverse phase HPLC, equipped with a flow through radioscanner. The results indicate that the intact epidermis metabolized 14C-18:2n6 to a group of novel products more polar than 18:2n6. In subsequent experiments, 14C-18:2n6 was either incubated with the 800 xg supernatant, the 105,000 xg pellet or supernatant from GP epidermis. Metabolism of 18:2n6 by the high speed supernatant resulted in the generation of polar products with chromatographic properties of not greater than 2 double bonds. These results indicate that although the GP epidermis lacks the capacity to desaturate 18:2n6 to 18:3n6, it can convert dietary 18:2n6 into a group of novel polar metabolites via a cytosolic mediated process. The function of these metabolites in the GP integumentary system remains to be determined

  11. Generation of novel metabolites of dietary linoleic acid (18:2n6) by guinea pig epidermis

    Chapkin, R.S.; Ziboh, V.A.

    1986-03-05

    Although the authors have demonstrated the inability of rat and guinea pig (GP) skin enzyme preparations to desaturate 18:2n6 into gammalinolenic acid (18:3n6) using an in vitro microsomal system, the fate of this dietary essential fatty acid in the GP epidermis is unknown. To explore the fate of 18:2n6, intact tissue slices from GP epidermis were incubated with (1-/sup 14/C)18:2n6. After incubation, the extracted lipids were transesterified using methanolic-HCL. The fatty acid methyl esters were analyzed using a combination of (i) argentation TLC, scanned using a proportional TLC radioscanner, and (ii) reverse phase HPLC, equipped with a flow through radioscanner. The results indicate that the intact epidermis metabolized /sup 14/C-18:2n6 to a group of novel products more polar than 18:2n6. In subsequent experiments, /sup 14/C-18:2n6 was either incubated with the 800 xg supernatant, the 105,000 xg pellet or supernatant from GP epidermis. Metabolism of 18:2n6 by the high speed supernatant resulted in the generation of polar products with chromatographic properties of not greater than 2 double bonds. These results indicate that although the GP epidermis lacks the capacity to desaturate 18:2n6 to 18:3n6, it can convert dietary 18:2n6 into a group of novel polar metabolites via a cytosolic mediated process. The function of these metabolites in the GP integumentary system remains to be determined.

  12. Transcriptional Activation and Production of Tryptophan-Derived Secondary Metabolites in Arabidopsis Roots Contributes to the Defense against the Fungal Vascular Pathogen Verticillium Iongisporum

    Tim Iven; Wolfgang Dr(o)ge-Laser; Stefanie K(o)nig; Seema Singh; Susanna A.Braus-Stromeyer; Matthias Bischoff; Lutz F.Tietze; Gerhard H.Braus; Volker Lipka; Ivo Feussner

    2012-01-01

    The soil-borne fungal pathogen Verticillium Iongisporum causes vascular disease on Brassicaceae host plants such as oilseed rape.The fungus colonizes the root xylem and moves upwards to the foliage where disease symptoms become visible.Using Arabidopsis as a model for early gene induction,we performed root transcriptome analyses in response to hyphal growth immediately after spore germination and during penetration of the root cortex,respectively.Infected roots showed a rapid reprogramming of gene expression such as activation of transcription factors,stress-,and defense-related genes.Here,we focused on the highly coordinated gene induction resulting in the production of tryptophan-derived secondary metabolites.Previous studies in leaves showed that enzymes encoded by CYP81F2 and PEN2 (PENETRATION2) execute the formation of antifungal indole glucosinolate (IGS) metabolites.In Verticillium-infected roots.we found transcriptional activation of CYP81F2 and the PEN2 homolog PEL1 (PEN2-LIKE1),but no increase in antifungal IGS breakdown products.In contrast,indole-3-carboxylic acid (I3CA) and the phytoalexin camalexin accumulated in infected roots but only camalexin inhibited Verticillium growth in vitro.Whereas genetic disruption of the individual metabolic pathways leading to either camalexin or CYP81F2-dependent IGS metabolites did not alter Verticillium-induced disease symptoms,a cyp79b2 cyp79b3 mutant impaired in both branches resulted in significantly enhanced susceptibility.Hence,our data provide an insight into root-specific early defenses and suggest tryptophan-derived metabolites as active antifungal compounds against a vascular pathogen.

  13. Prey-induced changes in the accumulation of amino acids and phenolic metabolites in the leaves of Drosera capensis L.

    Kováčik, Jozef; Klejdus, Bořivoj; Stork, František; Hedbavny, Josef

    2012-04-01

    Effect of prey feeding (ants Formica fusca) on the quantitative changes in the accumulation of free amino acids, soluble proteins, phenolic metabolites and mineral nutrients in the leaves of carnivorous plant Drosera capensis was studied. Arginine was the most abundant compound in Drosera leaves, while proline was abundant in ants. The amount of the majority of amino acids and their sum were elevated in the fed leaves after 3 and 21 days, and the same, but with further enhancement after 21 days, was observed in ants. Accumulation of amino acids also increased in young non-fed leaves of fed plants. Soluble proteins decreased in ants, but were not enhanced in fed leaves. This confirms the effectiveness of sundew's enzymatic machinery in digestion of prey and suggests that amino acids are not in situ deposited, but rather are allocated within the plant. The content of total soluble phenols, flavonoids and two selected flavonols (quercetin and kaempferol) was not affected by feeding in Drosera leaves, indicating that their high basal level was sufficient for the plant's metabolism and prey-induced changes were mainly N based. The prey also showed to be an important source of other nutrients besides N, and a stimulation of root uptake of some mineral nutrients is assumed (Mg, Cu, Zn). Accumulation of Ca and Na was not affected by feeding. PMID:21140278

  14. Chiral quizalofop-ethyl and its metabolite quizalofop-acid in soils: Enantioselective degradation, enzymes interaction and toxicity to Eisenia foetida.

    Ma, Lin; Liu, Hui; Qu, Han; Xu, Yangguang; Wang, Peng; Sun, Mingjing; Zhou, Zhiqiang; Liu, Donghui

    2016-06-01

    An enantioselective chromatographic method to analyze enantiomers of quizalofop-ethyl and its metabolite quizalofop-acid was established using a high-performance liquid chromatography (HPLC) on (R, R) Whelk-O 1 column. The enantioselective degradation kinetics of quizalofop-ethyl and quizalofop-acid in three soils were investigated. Moreover, the interaction with urease and catalase in the soils and the acute toxicity to Eisenia foetida of quizalofop-ethyl were also determined in order to assess their metabolism mechanism and environmental risk. From the results, quizalofop-ethyl was configurationally stable and was hydrolyzed rapidly to quizalofop-acid, which also degraded enantioselectively but slowly, and the inversion of the S-(-)-quizalofop-acid into the R-(+)-quizalofop-acid was observed in Xinxiang soil. In addition, quizalofop-ethyl and quizalofop-acid enantioselectively affected urease activity but not catalase. The acute toxicity assays to earthworm indicated that the racemic quizalofop-ethyl and quizalofop-acid were more toxic than quizalofop-p-ethyl and quizalofop-p-acid respectively, dramatically, the toxicity of the metabolite was much higher than the parent compound. These results revealed the enantioselective degradation of quizalofop-ethyl and quizalofop-acid, and the differences of toxicity among the enantiomers of the parent compound and the metabolite, which should be considered in future environmental risk evaluation. PMID:26971169

  15. Metabolite changes during natural and lactic acid bacteria fermentations in pastes of soybeans and soybean-maize blends.

    Ng'ong'ola-Manani, Tinna Austen; Ostlie, Hilde Marit; Mwangwela, Agnes Mbachi; Wicklund, Trude

    2014-11-01

    The effect of natural and lactic acid bacteria (LAB) fermentation processes on metabolite changes in pastes of soybeans and soybean-maize blends was studied. Pastes composed of 100% soybeans, 90% soybeans and 10% maize, and 75% soybeans and 25% maize were naturally fermented (NFP), and were fermented by lactic acid bacteria (LFP). LAB fermentation processes were facilitated through back-slopping using a traditional fermented gruel, thobwa as an inoculum. Naturally fermented pastes were designated 100S, 90S, and 75S, while LFP were designated 100SBS, 90SBS, and 75SBS. All samples, except 75SBS, showed highest increase in soluble protein content at 48 h and this was highest in 100S (49%) followed by 90SBS (15%), while increases in 100SBS, 90S, and 75S were about 12%. Significant (P Lactic acid was 2.5- to 3.5-fold higher in LFP than in NFP, and other organic acids detected were acetate and succinate. Maltose levels were the highest among the reducing sugars and were two to four times higher in LFP than in NFP at the beginning of the fermentation, but at 72 h, only fructose levels were significantly (P Enzyme activities were higher in LFP at 0 h, but at 72 h, the enzyme activities were higher in NFP. Both fermentation processes improved nutritional quality through increased protein and amino acid solubility and degradation of phytic acid (85% in NFP and 49% in LFP by 72 h). PMID:25493196

  16. Valorization of biodiesel derived glycerol as a carbon source to obtain added-value metabolites: Focus on polyunsaturated fatty acids.

    Abad, Sergi; Turon, Xavier

    2012-01-01

    The amount of glycerol derived from the biodiesel industry is exponentially increasing. The valorization of glycerol has acquired attention and resources with an obvious economic and environmental interest. Glycerol has the potential to improve the profitability of biodiesel in a biorefinery scenario. Added-value metabolites obtained from glycerol-based fermentations are the target of multiple research studies, primarily chemicals and biopolymers. Pigments and polyunsaturated fatty acids are exceptional examples as they have market presence as nutraceuticals. Most of the studies reviewed have been based on microalgae cultures. Depending on the strain and the engineering aspects of such cultures the final yield suffers notable variations. This is an emerging field which shows great potential from the perspective of a byproduct usage and the increasing yields (value) obtained from the bioprocess. PMID:22261015

  17. The shikimic acid: an important metabolite for the Aglianico del Vulture wines

    Pasquale Tamborra

    2014-12-01

    Full Text Available Shikimic acid is a precursor for the biosynthesis of aromatic amino acids and flavonoids (anthocyanins, tannins and flavonols. In the pharmaceutical industry, it is obtained by extraction of star anise from China, and at a yield of 3-7% it is used for the production of antiviral drug, e.g. oseltamivir. Unlike flavonoids which are only present in the grape skins, shikimic acid is present in the juice together with hydroxycinnamil tartaric acids (caffeic, ferulic and p-coumaric acid. Therefore, their content in white wines may not be negligible and their presence may explain the epidemiological studies that showed a reduced incidence of cardiovascular diseases also in people with moderate white wine consumption. The content of shikimic acid has been used to characterize wines. In southern Italy it has been used to distinguish Aglianico grape, which holds medium-high content, from Negroamaro, Primitivo and Uva di Troia grapes who have rather lower levels. It could be useful also to distinguish Fiano di Avellino (high value from Fiano Minutolo (low value. However, results of a recent work showed that the shikimic acid content decreases significantly during the ripening of the grapes and therefore its content in wine is strongly influenced by the harvest period. Finally, in a recent paper it was highlighted the increase in shikimic acid content at the end of fermentation in an Aglianico del Vulture wine, produced in the area of Rapolla (PZ, Italy municipality during the 2013 harvest. These last experimental results explain why the values of shikimic acid were lower in grapes and surprisingly higher in wines produced in the 2011 and 2012 harvest.

  18. Simultaneous quantitation of acetylsalicylic acid and clopidogrel along with their metabolites in human plasma using liquid chromatography tandem mass spectrometry.

    Chhonker, Yashpal S; Pandey, Chandra P; Chandasana, Hardik; Laxman, Tulsankar Sachin; Prasad, Yarra Durga; Narain, V S; Dikshit, Madhu; Bhatta, Rabi S

    2016-03-01

    The interest in therapeutic drug monitoring has increased over the last few years. Inter- and intra-patient variability in pharmacokinetics, plasma concentration related toxicity and success of therapy have stressed the need of frequent therapeutic drug monitoring of the drugs. A sensitive, selective and rapid liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) method was developed for the simultaneous quantification of acetylsalicylic acid (aspirin), salicylic acid, clopidogrel and carboxylic acid metabolite of clopidogrel in human plasma. The chromatographic separations were achieved on Waters Symmetry Shield(TM) C18 column (150 × 4.6 mm, 5 µm) using 3.5 mm ammonium acetate (pH 3.5)-acetonitrile (10:90, v/v) as mobile phase at a flow rate of 0.75 mL/min. The present method was successfully applied for therapeutic drug monitoring of aspirin and clopidogrel in 67 patients with coronary artery disease. PMID:26230053

  19. Anti-inflammatory activity of mushrooms extracts, identified phenolic acids and their possible metabolites

    Taofiq, Oludemi; Calhelha, Ricardo C.; Heleno, Sandrina A.; Barros, Lillian; Martins, Anabela; Abreu, Rui M.V.; Queiroz, Maria João R. P.; Isabel C. F. R. Ferreira

    2015-01-01

    Mushrooms are rich sources of many bioactive compounds, such as phenolic acids, that play an important role in the organism, acting as antioxidants, antitumors, antimicrobials, immunomodulators, among others. However, their anti-inflammatory activity has not been deeply studied. In the present study, the ethanolic extracts of fourteen edible mushroom species were firstly characterized in terms of phenolic acids and related compounds by HPLC-PDA, followed by the study of the ant...

  20. Metabolomics-based prediction models of yeast strains for screening of metabolites contributing to ethanol stress tolerance

    Hashim, Z.; Fukusaki, E.

    2016-06-01

    The increased demand for clean, sustainable and renewable energy resources has driven the development of various microbial systems to produce biofuels. One of such systems is the ethanol-producing yeast. Although yeast produces ethanol naturally using its native pathways, production yield is low and requires improvement for commercial biofuel production. Moreover, ethanol is toxic to yeast and thus ethanol tolerance should be improved to further enhance ethanol production. In this study, we employed metabolomics-based strategy using 30 single-gene deleted yeast strains to construct multivariate models for ethanol tolerance and screen metabolites that relate to ethanol sensitivity/tolerance. The information obtained from this study can be used as an input for strain improvement via metabolic engineering.

  1. Detection of pharmaceuticals and phytochemicals together with their metabolites in hospital effluents in Japan, and their contribution to sewage treatment plant influents.

    Azuma, Takashi; Arima, Natsumi; Tsukada, Ai; Hirami, Satoru; Matsuoka, Rie; Moriwake, Ryogo; Ishiuchi, Hirotaka; Inoyama, Tomomi; Teranishi, Yusuke; Yamaoka, Misato; Mino, Yoshiki; Hayashi, Tetsuya; Fujita, Yoshikazu; Masada, Mikio

    2016-04-01

    The occurrence of 41 pharmaceuticals and phytochemicals (PPs) including their metabolites was surveyed in hospital effluent in an urban area of Japan. A detailed survey of sewage treatment plant (STP) influent and effluent, and river water was also conducted. Finally, mass balances with mass fluxes of the target PPs through the water flow were evaluated and the degree of contribution of hospital effluent to the environmental discharge was estimated. The results indicate that 38 compounds were detectable in hospital effluent over a wide concentration range from ng/L to μg/L, with a maximum of 92μg/L. The contributions of PPs in the hospital effluent to STP influent varied widely from waters. PMID:26802347

  2. Altered secretion of selected arachidonic acid metabolites during subclinical endometritis relative to estrous cycle stage and grade of fibrosis in mares.

    Gajos, Katarzyna; Kozdrowski, Roland; Nowak, Marcin; Siemieniuch, Marta J

    2015-08-01

    Mares that fail to become pregnant after repeated breeding, without showing typical signs of clinical endometritis, should be suspected of subclinical endometritis (SE). Contact with infectious agents results in altered synthesis and secretion of inflammatory mediators, including cytokines and arachidonic acid metabolites, and disturbs endometrial functional balance. To address the hypothesis that SE affects the immune endocrine status of the equine endometrium, spontaneous secretion of prostaglandin E(2) (PGE(2)), prostaglandin F(2α) (PGF(2α)), 6-keto-PGF(1α )(a metabolite of prostacyclin I(2)), leukotriene B(4) (LTB(4)), and leukotriene C(4) (LTC(4)) was examined. In addition, secretion of these factors was examined relative to the grade of inflammation, fibrosis, and estrous cycle stage. Eighty-two warmblood mares, of known breeding history, were enrolled in this study. On the basis of histopathologic assessment, mares were classified as suffering from first-grade SE, second-grade SE, or being healthy. The grade of fibrosis and the infiltration of endometrial tissue with polymorphonuclear leukocytes were examined by routine hematoxylin-eosin staining. In mares suffering from SE, the secretion profiles of PGE(2), 6-keto-PGF(1α), LTB(4), and LTC(4) were changed compared to mares that did not suffer from endometritis. The secretion of PGE(2) and 6-keto-PGF1α was increased, whereas that of LTB(4) and LTC(4) was decreased. Secretion of 6-keto-PGF(1α) was increased in first- and second-grade SE (P < 0.01). The concentration of PGI(2) metabolite was increased only in inflamed endometrium, independently of the inflammation grade, but was not affected by fibrosis. Prostaglandin E(2) secretion was increased in second-grade SE (P < 0.05). The secretion of LTB(4) decreased in both first- and second-grade SE (P < 0.05), whereas secretion of LTC(4) was decreased only in second-grade SE (P < 0.05). Fibrosis did not change the secretion profile of PGE(2), PGF(2α), and 6

  3. Docosapentaenoic acid derived metabolites and mediators - The new world of lipid mediator medicine in a nutshell.

    Weylandt, Karsten-H

    2016-08-15

    Recent years have seen the description and elucidation of a new class of anti-inflammatory and pro-resolving lipid mediators. The arachidonic acid (AA)-derived compounds in this class are called lipoxins and have been described in great detail since their discovery thirty years ago. The new players are mediators derived from fish oil omega-3 fatty acids eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), called resolvins, protectins and maresins. Taken together, these mediators are also called specialized pro-resolution mediators (SPMs). As compared to the AA/EPA/DHA-derived compounds, research regarding mediators formed from the n-3 and n-6 docosapentaenoic acids (DPAn-3 and DPAn-6) is sparse. However, mono- di- and trihydroxy derivates of the DPAs have anti-inflammatory properties as well, even though mechanisms of their anti-inflammatory action have not been fully elucidated. This review aims to summarize current knowledge regarding the DPA-derived SPMs and their actions. PMID:26546723

  4. Probing the molecular and electronic structure of the lichen metabolite usnic acid: A DFT study

    Graphical abstract: DFT calculations of structural preferences, acidic properties, carbonyl vibrations, 13C NMR chemical shifts, and absorption spectrum account for the unique structural backbone, chemical behaviour, and spectroscopic properties of usnic acid, the cortical pigment and potent reactive of lichens. - Abstract: The molecular structure of usnic acid was investigated by the density functional theory (DFT). Two keto-enol tautomers are nearly isoenergetic and more stable than other tautomers. Noteworthy is the energy difference among the three intramolecular O-H...O hydrogen bonds. The DFT/PCM calculated dissociation constants account for the acidic sequence of the three OH-groups. The electronic structure was also studied by calculating IR/Raman, NMR, and absorption features. A reliable assignment of the 'fingerprint' carbonyl stretching modes was supported by calculations on related molecules. The calculated NMR chemical shifts fit expectation in terms of a fast interconversion between the two most preferred tautomers. A variety of π → π* and n → π* excitations, localized on a single ring or involving a charge-transfer between the two lateral rings of the molecule, gives rise to the broad UV-absorption bands. This property accounts for the efficient protection against damaging solar radiation provided by usnic acid for lichens.

  5. Effects of chicory inulin on serum metabolites of uric acid, lipids, glucose, and abdominal fat deposition in quails induced by purine-rich diets.

    Lin, Zhijian; Zhang, Bing; Liu, Xiaoqing; Jin, Rui; Zhu, Wenjing

    2014-11-01

    Inulin, a group of dietary fibers, is reported to improve the metabolic disorders. In the present study, we investigated the effects of chicory inulin on serum metabolites of uric acid (UA), lipids, glucose, and abdominal fat deposition in quail model induced by a purine-rich diet. In this study, 60 male French quails were randomly allocated to five groups: CON (control group), MOD (model group), BEN (benzbromarone-treated group), CHI-H (high-dosage chicory inulin-treated group), and CHI-L (low-dosage chicory inulin-treated group). The serum UA level was significantly increased in the model group from days 7 to 28, as well as triglyceride (TG) and free fatty acid (FFA) increased later in the experimental period. The abdominal fat ratio was increased on day 28. Benzbromarone can decrease UA levels on days 14 and 28. The high and low dosage of chicory inulin also decreased serum UA levels on days 7, 14, and 28. The abdominal fat ratio, activity, and protein of acetyl-CoA carboxylase (ACC) were decreased in chicory inulin-treated groups. The activities of xanthine oxidase (XOD) and fatty acid synthase (FAS) were increased in the model group and decreased in the benzbromarone and chicory inulin groups. This study evaluated a quail model of induced hyperuricemia with other metabolic disorders caused by a high-purine diet. The results indicated that a purine-rich diet might contribute to the development of hyperuricemia, hypertriglyceridemia, and abdominal obesity. Chicory inulin decreased serum UA, TG, and abdominal fat deposition in a quail model of hyperuricemia by altering the ACC protein expression and FAS and XOD activities. PMID:25314375

  6. Tissue-specific activation of mitogen-activated protein kinases for expression of transthyretin by phenylalanine and its metabolite, phenylpyruvic acid

    Park, Joo-Won; Lee, Mi Hee; Choi, Jin-Ok; Park, Hae-Young; Jung, Sung-Chul

    2009-01-01

    Phenylketonuria is an autosomal recessive disorder caused by a deficiency of phenylalanine hydroxylase. Transthyretin has been implicated as an indicator of nutritional status in phenylketonuria patients. In this study, we report that phenylalanine and its metabolite, phenylpyruvic acid, affect MAPK, changing transthyretin expression in a cell- and tissue-specific manner. Treatment of HepG2 cells with phenylalanine or phenylpyruvic acid decreased transcription of the TTR gene and decreased th...

  7. The cerebral metabolism of amino acids and related metabolites as studied by {sup 13}C and {sup 14}C labelling

    Hassel, B.

    1995-11-01

    The present investigations show the feasibility of analyzing the cerebral metabolism of amino acids and related metabolites by {sup 13}C-and {sup 14}C-labelling using labelled acetate and glucose as markers for glial and neuronal metabolism, respectively. Using [{sup 13}C]acetate, it was shown that glial cells export {approx}60% of their TCA cycle intermediates, mostly as glutamine, and that this glutamine is used by neurons partly as an energy reserve, and partly it is converted directly to glutamate and GABA. Using [{sup 13}C]glucose, the glial process or pyruvate carboxylation was shown to compensate fully for the loss of glutamine. The mechanism of action of two neurotoxins, fluorocitrate and 3-nitropropionate was elucidated. The latter toxin was shown to inhibit the TCA cycle of GABAergic neurons selectively. Formation of pyruvate and lactate from glial TCA cycle intermediates was demonstrated in vivo. This pathway may be important for glial inactivation of transmitter glutamate and GABA. The results illustrate glianeuronal interactions, and they suggest the applicability of {sup 13}CNMR spectroscopy to the detailed study of the cerebral metabolism of amino acids in the intact, unanesthetized human brain. 174 refs.

  8. The cerebral metabolism of amino acids and related metabolites as studied by 13C and 14C labelling

    The present investigations show the feasibility of analyzing the cerebral metabolism of amino acids and related metabolites by 13C-and 14C-labelling using labelled acetate and glucose as markers for glial and neuronal metabolism, respectively. Using [13C[acetate, it was shown that glial cells export ∼60% of their TCA cycle intermediates, mostly as glutamine, and that this glutamine is used by neurons partly as an energy reserve, and partly it is converted directly to glutamate and GABA. Using [13C[glucose, the glial process or pyruvate carboxylation was shown to compensate fully for the loss of glutamine. The mechanism of action of two neurotoxins, fluorocitrate and 3-nitropropionate was elucidated. The latter toxin was shown to inhibit the TCA cycle of GABAergic neurons selectively. Formation of pyruvate and lactate from glial TCA cycle intermediates was demonstrated in vivo. This pathway may be important for glial inactivation of transmitter glutamate and GABA. The results illustrate glianeuronal interactions, and they suggest the applicability of 13CNMR spectroscopy to the detailed study of the cerebral metabolism of amino acids in the intact, unanesthetized human brain. 174 refs

  9. INCORPORATION OF OMEGA-3 FATTY ACID AND SYNTHESIS OF THEIR METABOLITES IN MUSCLE TISSUE IN MICE FED WITH FLAXSEED AND PERILLA-ENRICHED DIET

    Sheisa Cyleia Sargi

    2013-01-01

    Full Text Available Polyunsaturated fatty acids omega 3 and omega 6 are believed to have beneficial physiological effects on the human body. The present study aimed to evaluate the incorporation and synthesis of omega-3 fatty acid and their metabolites in muscle tissue in mice fed with a flaxseed and perilla-enriched diet. The animals were fed a diet supplemented with flaxseed and perilla meals for fifty-six days and control animals received a commercial diet. Mice were sacrificed on the 7th, 28th and 56th days and the muscle tissue was collected. The FA concentration was analyzed by gas chromatography. The levels of alpha-Linolenic Acid (LNA and their metabolites, Eicosapentaenoic Acid (EPA and Docosahexaenoic Acid (DHA increased as a function of feeding length in the groups given the supplemented diet. LNA was incorporated into muscle tissue and conversion into Very Long Chain Polyunsaturated Fatty Acids (VLC-PUFA was observed. The muscle tissue of animals fed with enriched diets presented a high sum of n-3 FA and high concentrations of LNA, EPA and DHA, as compared to mice fed commercial diet. The intake of flaxseed and perilla-meal enriched diets by mice may result in the deposition of LNA and its metabolites EPA and DHA into muscle tissue.

  10. Do the metabolites of 6-[F-18]fluoro-L-dopa and of [F-18]fluoro-meta-L-tyrosine contribute to the F-18 accumulation in the human brain?

    The purpose of this study was to determine if the metabolites of 6-[F-18]fluoro-L-dopa (F-dopa) and of [F-18]fluoro-meta-L-tyrosine (FmLtyr) contribute to the accumulation of fluorine-18 in the brain through unspecific retention. PET studies were conducted on a healthy human subject who was treated with both of the radiopharmaceuticals and their labelled metabolites. Results indicated that in contrast to F-dopa, the metabolite of FmLtyr does not 'contaminate' the brain with extraneous fluorine-18

  11. Linoleic acid metabolite drives severe asthma by causing airway epithelial injury

    Mabalirajan, Ulaganathan; Rehman, Rakhshinda; Ahmad, Tanveer; Kumar, Sarvesh; Singh, Suchita; Leishangthem, Geeta D.; Aich, Jyotirmoi; Kumar, Manish; Khanna, Kritika; Singh, Vijay P.; Dinda, Amit K; Biswal, Shyam; Agrawal, Anurag; Ghosh, Balaram

    2013-01-01

    Airway epithelial injury is the hallmark of various respiratory diseases, but its mechanisms remain poorly understood. While 13-S-hydroxyoctadecadienoic acid (13-S-HODE) is produced in high concentration during mitochondrial degradation in reticulocytes little is known about its role in asthma pathogenesis. Here, we show that extracellular 13-S-HODE induces mitochondrial dysfunction and airway epithelial apoptosis. This is associated with features of severe airway obstruction, lung remodeling...

  12. Biological Significance of Urolithins, the Gut Microbial Ellagic Acid-Derived Metabolites: The Evidence So Far

    Juan Carlos Espín; Mar Larrosa; María Teresa García-Conesa; Francisco Tomás-Barberán

    2013-01-01

    The health benefits attributed to pomegranate have been associated with its high content in polyphenols, particularly ellagitannins. This is also the case for other ellagitannin-containing fruits and nuts including strawberry, raspberry, blackberry, walnuts, and muscadine grapes. The bioavailability of ellagitannins and ellagic acid is however very low. These molecules suffer extensive metabolism by the gut microbiota to produce urolithins that are much better absorbed. Urolithins circulate i...

  13. Targeted Chiral Analysis of Bioactive Arachidonic Acid Metabolites Using Liquid-Chromatography-Mass Spectrometry

    Clementina Mesaros

    2012-04-01

    Full Text Available A complex structurally diverse series of eicosanoids arises from the metabolism of arachidonic acid. The metabolic profile is further complicated by the enantioselectivity of eicosanoid formation and the variety of regioisomers that arise. In order to investigate the metabolism of arachidonic acid in vitro or in vivo, targeted methods are advantageous in order to distinguish between the complex isomeric mixtures that can arise by different metabolic pathways. Over the last several years this targeted approach has become more popular, although there are still relatively few examples where chiral targeted approaches have been employed to directly analyze complex enantiomeric mixtures. To efficiently conduct targeted eicosanoid analyses, LC separations are coupled with collision induced dissociation (CID and tandem mass spectrometry (MS/MS. Product ion profiles are often diagnostic for particular regioisomers. The highest sensitivity that can be achieved involves the use of selected reaction monitoring/mass spectrometry (SRM/MS; whereas the highest specificity is obtained with an SRM transitions between an intense parent ion, which contains the intact molecule (M and a structurally significant product ion. This review article provides an overview of arachidonic acid metabolism and targeted chiral methods that have been utilized for the analysis of the structurally diverse eicosanoids that arise.

  14. Effect of polyunsaturated fatty acids and their metabolites on bleomycin-induced cytotoxic action on human neuroblastoma cells in vitro.

    Sailaja Polavarapu

    Full Text Available In the present study, we noted that bleomycin induced growth inhibitory action was augmented by all the polyunsaturated fatty acids (PUFAs tested on human neuroblastoma IMR-32 (0.5 × 10(4 cells/100 µl of IMR cells (EPA > DHA > ALA = GLA = AA > DGLA = LA: ∼ 60, 40, 30, 10-20% respectively at the maximum doses used. Of all the prostaglandins (PGE1, PGE2, PGF2α, and PGI2 and leukotrienes (LTD4 and LTE4 tested; PGE1, PGE2 and LTD4 inhibited the growth of IMR-32 cells to a significant degree at the highest doses used. Lipoxin A4 (LXA4, 19,20-dihydroxydocosapentaenoate (19, 20 DiHDPA and 10(S,17(S-dihydroxy-4Z,7Z,11E,13Z,15E,19Z-docosahexaenoic acid (protectin: 10(S,17(SDiHDoHE, metabolites of DHA, significantly inhibited the growth of IMR-32 cells. Pre-treatment with AA, GLA, DGLA and EPA and simultaneous treatment with all PUFAs used in the study augmented growth inhibitory action of bleomycin. Surprisingly, both indomethacin and nordihydroguaiaretic acid (NDGA at 60 and 20 µg/ml respectively enhanced the growth of IMR-32 cells even in the presence of bleomycin. AA enhanced oxidant stress in IMR-32 cells as evidenced by an increase in lipid peroxides, superoxide dismutase levels and glutathione peroxidase activity. These results suggest that PUFAs suppress growth of human neuroblastoma cells, augment growth inhibitory action of bleomycin by enhancing formation of lipid peroxides and altering the status of anti-oxidants and, in all probability, increase the formation of lipoxins, resolvins and protectins from their respective precursors that possess growth inhibitory actions.

  15. The highly abundant urinary metabolite urobilin interferes with the bicinchoninic acid assay.

    Sampson, D L; Chng, Y L; Upton, Z; Hurst, C P; Parker, A W; Parker, T J

    2013-11-01

    Estimation of total protein concentration is an essential step in any protein- or peptide-centric analysis pipeline. This study demonstrates that urobilin, a breakdown product of heme and a major constituent of urine, interferes considerably with the bicinchoninic acid (BCA) assay. This interference is probably due to the propensity of urobilin to reduce cupric ions (Cu(2+)) to cuprous ions (Cu(1+)), thus mimicking the reduction of copper by proteins, which the assay was designed to do. In addition, it is demonstrated that the Bradford assay is more resistant to the influence of urobilin and other small molecules. As such, urobilin has a strong confounding effect on the estimate of total protein concentrations obtained by BCA assay and thus this assay should not be used for urinary protein quantification. It is recommended that the Bradford assay be used instead. PMID:23911526

  16. NMDA Receptors and Oxidative Stress Induced by the Major Metabolites Accumulating in HMG Lyase Deficiency Mediate Hypophosphorylation of Cytoskeletal Proteins in Brain From Adolescent Rats: Potential Mechanisms Contributing to the Neuropathology of This Disease.

    Fernandes, Carolina Gonçalves; Pierozan, Paula; Soares, Gilberto Machado; Ferreira, Fernanda; Zanatta, Ângela; Amaral, Alexandre Umpierrez; Borges, Clarissa Günther; Wajner, Moacir; Pessoa-Pureur, Regina

    2015-10-01

    Neurological symptoms and cerebral abnormalities are commonly observed in patients with 3-hydroxy-3-methylglutaryl-CoA lyase (HMG lyase) deficiency, which is biochemically characterized by predominant tissue accumulation of 3-hydroxy-3-methylglutaric (HMG), 3-methylglutaric (MGA), and 3-methylglutaconic (MGT) acids. Since the pathogenesis of this disease is poorly known, the present study evaluated the effects of these compounds on the cytoskeleton phosphorylating system in rat brain. HMG, MGA, and MGT caused hypophosphorylation of glial fibrillary acidic protein (GFAP) and of the neurofilament subunits NFL, NFM, and NFH. HMG-induced hypophosphorylation was mediated by inhibiting the cAMP-dependent protein kinase (PKA) on Ser55 residue of NFL and c-Jun kinase (JNK) by acting on KSP repeats of NFM and NFH subunits. We also evidenced that the subunit NR2B of NMDA receptor and Ca(2+) was involved in HMG-elicited hypophosphorylation of cytoskeletal proteins. Furthermore, the antioxidants L-NAME and TROLOX fully prevented both the hypophosphorylation and the inhibition of PKA and JNK caused by HMG, suggesting that oxidative damage may underlie these effects. These findings indicate that the main metabolites accumulating in HMG lyase deficiency provoke hypophosphorylation of cytoskeleton neural proteins with the involvement of NMDA receptors, Ca(2+), and reactive species. It is presumed that these alterations may contribute to the neuropathology of this disease. PMID:26174040

  17. Microfluidic study of the chemotactic response of Escherichia coli to amino acids, signaling molecules and secondary metabolites.

    Nagy, Krisztina; Sipos, Orsolya; Valkai, Sándor; Gombai, Éva; Hodula, Orsolya; Kerényi, Ádám; Ormos, Pál; Galajda, Péter

    2015-07-01

    Quorum sensing and chemotaxis both affect bacterial behavior on the population level. Chemotaxis shapes the spatial distribution of cells, while quorum sensing realizes a cell-density dependent gene regulation. An interesting question is if these mechanisms interact on some level: Does quorum sensing, a density dependent process, affect cell density itself via chemotaxis? Since quorum sensing often spans across species, such a feedback mechanism may also exist between multiple species. We constructed a microfluidic platform to study these questions. A flow-free, stable linear chemical gradient is formed in our device within a few minutes that makes it suitable for sensitive testing of chemoeffectors: we showed that the amino acid lysine is a weak chemoattractant for Escherichia coli, while arginine is neutral. We studied the effect of quorum sensing signal molecules of Pseudomonas aeruginosa on E. coli chemotaxis. Our results show that N-(3-oxododecanoyl)-homoserine lactone (oxo-C12-HSL) and N-(butryl)-homoserine lactone (C4-HSL) are attractants. Furthermore, we tested the chemoeffector potential of pyocyanin and pyoverdine, secondary metabolites under a quorum sensing control. Pyocyanin is proved to be a weak attractant while pyoverdine are repellent. We demonstrated the usability of the device in co-culturing experiments, where we showed that various factors released by P. aeruginosa affect the dynamic spatial rearrangement of a neighboring E. coli population, while surface adhesion of the cells is also modulated. PMID:26339306

  18. Interactions of valproic acid with carbamazepine and its metabolites' concentrations, concentrations ratios, and level/dose ratios in epileptic children.

    Liu, H; Delgado, M R; Browne, R H

    1995-02-01

    In two groups of epileptic children receiving carbamazepine (CBZ) therapy with or without valproic acid (VPA) comedication, we investigate the drug interactions of VPA on serum CBZ and its metabolites' concentrations, concentration ratios, and level/dose ratios. Serum total and free CBZ-10, 11-epoxide (CBZ-E) concentrations are significantly increased in patients taking CBZ plus VPA, together with higher CBZ-E/CBZ concentration ratios and CBZ-E level/dose ratios. These results reflect the accumulation of CBZ-E. The decreased concentration ratios of trans-10, 11-dihydroxy-10, 11-dihydro-CBZ (CBZ-H)/CBZ-E observed in patients taking CBZ plus VPA suggest an inhibition in the biotransformation from CBZ-E to CBZ-H. Significant negative correlations are found between serum VPA level and CBZ-H/CBZ-E concentration ratios, indicating that the inhibition of CBZ-E hydrolysis by VPA may depend on the concentration of VPA (total or free CBZ-H/CBZ-E concentration ratio = [formula: see text], respectively). VPA concentration also shows significant positive correlations with CBZ-E and CBZ level/dose ratios. Patients taking CBZ plus VPA have significant higher free fractions of CBZ and CBZ-E than do patients on CBZ alone, suggesting a protein-binding displacement by VPA. PMID:8665529

  19. Mutagenicity and DNA-damaging potential of clenbuterol and its metabolite 4-amino-3,5-dichlorobenzoic acid in vitro.

    Vulić, Ana; Durgo, Ksenija; Pleadin, Jelka; Herceg, Luka; Kopjar, Nevenka

    2015-03-01

    The aim of this study was to evaluate in vitro toxicity of clenbuterol and its metabolite 4-amino-3,5-dichlorobenzoic acid. Cytotoxicity and pro-oxidative effect of both compounds were studied on human colon adenocarcinoma cell line SW 480. No significant cytotoxic effect of either compound was observed. Results of an Ames test on Salmonella typhimurium did not indicate mutagenic activity of clenbuterol on TA 98 and TA 100 strains, regardless of metabolic activation. Potential mutagenic effects of the highest clenbuterol concentration (2500 ng/ml) were observed on the TA 1535 strain. The obtained results of alkaline comet assay on isolated human lymphocytes suggested that both compounds induced an increase of primary DNA damage in a concentration-dependent manner. 4-ADBA was a slightly more potent inducer of primary DNA damage as compared to clenbuterol. Chromosomal aberration analysis showed that clenbuterol caused a statistically significant increase in the total number of aberrant cells only at the highest concentration tested (3% vs. 0.7% in the negative control). The results of this study might represent a solid frame for designing and planning future studies with both compounds, which should further clarify their mechanisms of action and genotoxic/cytogenetic effects relevant for human risk assessment. PMID:25595371

  20. Melatonin promotes Bax sequestration to mitochondria reducing cell susceptibility to apoptosis via the lipoxygenase metabolite 5-hydroxyeicosatetraenoic acid

    Radogna, Flavia

    2015-03-01

    Extra-neurological functions of melatonin include control of the immune system and modulation of apoptosis. We previously showed that melatonin inhibits the intrinsic apoptotic pathway in leukocytes via stimulation of high affinity MT1/MT2 receptors, thereby promoting re-localization of the anti-apoptotic Bcl-2 protein to mitochondria. Here we show that Bcl-2 sequesters pro-apoptotic Bax into mitochondria in an inactive form after melatonin treatment, thus reducing cell propensity to apoptosis. Bax translocation and the anti-apoptotic effect of melatonin are strictly dependent on the presence of Bcl-2, and on the 5-lipoxygenase (5-LOX) metabolite 5-hydroxyeicosatetraenoic acid (5-HETE), which we have previously shown to be produced as a consequence of melatonin binding to its low affinity target calmodulin. Therefore, the anti-apoptotic effect of melatonin requires the simultaneous, independent interaction with high (MT1/MT2) and low (calmodulin) affinity targets, eliciting two independent signal transduction pathways converging into Bax sequestration and inactivation. MT1/MT2 vs. lipoxygenase pathways are activated by 10-9 vs. 10-5M melatonin, respectively; the anti-apoptotic effect of melatonin is achieved at 10-5M, but drops to 10-9M upon addition of exogenous 5-HETE, revealing that lipoxygenase activation is the rate-limiting pathway. Therefore, in areas of inflammation with increased 5-HETE levels, physiological nanomolar concentrations of melatonin may suffice to maintain leukocyte viability.

  1. Development and Validation of an HPLC Method for Simultaneous Quantification of Clopidogrel Bisulfate, Its Carboxylic Acid Metabolite, and Atorvastatin in Human Plasma: Application to a Pharmacokinetic Study

    Octavian Croitoru

    2015-01-01

    Full Text Available A simple, sensitive, and specific reversed phase liquid chromatographic method was developed and validated for simultaneous quantification of clopidogrel, its carboxylic acid metabolite, and atorvastatin in human serum. Plasma samples were deproteinized with acetonitrile and ibuprofen was chosen as internal standard. Chromatographic separation was performed on an BDS Hypersil C18 column (250 × 4.6 mm; 5 μm via gradient elution with mobile phase consisting of 10 mM phosphoric acid (sodium buffer solution (pH = 2.6 adjusted with 85% orthophosphoric acid : acetonitrile : methanol with flow rate of 1 mL·min−1. Detection was achieved with PDA detector at 220 nm. The method was validated in terms of linearity, sensitivity, precision, accuracy, limit of quantification, and stability tests. Calibration curves of the analytes were found to be linear in the range of 0.008–2 μg·mL−1 for clopidogrel, 0.01–4 μg·mL−1 for its carboxylic acid metabolite, and 0.005–2.5 μg·mL−1 for atorvastatin. The results of accuracy (as recovery with ibuprofen as internal standard were in the range of 96–98% for clopidogrel, 94–98% for its carboxylic acid metabolite, and 90–99% for atorvastatin, respectively.

  2. Structure-dependent activities of polybrominated diphenyl ethers and hydroxylated metabolites on zebrafish retinoic acid receptor.

    Zhao, Jing; Zhu, Xiangwei; Xu, Ting; Yin, Daqiang

    2015-02-01

    Polybrominated diphenyl ethers (PBDEs), a group of potential endocrine-disrupting chemicals (EDCs) have been shown to disrupt retinoid homeostasis in different species in both laboratory and field studies. However, the molecular mechanisms of interactions with the retinoic acid receptor (RAR) are not fully understood. Zebrafish have proven useful for investigating mechanisms of chemical toxicity. In the present study, a reporter gene assay was used to investigate the activities of 11 PBDEs and six OH-PBDEs with different degrees of bromination on zebrafish RAR. All tested OH-PBDEs induced RAR transcriptional activity; however, of the 11 PBDEs examined, only BDE28 and BDE154 affected the RAR transcriptional activity. Homology modeling and molecular docking were employed to simulate the interactions of PBDEs/OH-PBDEs with zebrafish RARs and to identify binding affinities to analyze the specialization of the interaction between RARs and PBDEs/OH-PBDEs. The results showed that although these compounds could bind with RARs, the effects of PBDEs/OH-PBDEs on RAR transcriptional activity did not depend on their RAR-binding abilities. The present study is the first attempt to demonstrate that OH-PBDEs could induce RAR transcriptional activity by binding directly with RAR; these effects are possibly related to the structure of the compounds, especially their hydroxylation and bromination. Most of the PBDEs could not directly interact with the RAR. PMID:25077655

  3. 10-oxo-12(Z)-octadecenoic acid, a linoleic acid metabolite produced by gut lactic acid bacteria, potently activates PPARγ and stimulates adipogenesis

    Goto, Tsuyoshi, E-mail: tgoto@kais.kyoto-u.ac.jp [Laboratory of Molecular Function of Food, Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Uji 611-0011 (Japan); Research Unit for Physiological Chemistry, The Center for the Promotion of Interdisciplinary Education and Research, Kyoto University (Japan); Kim, Young-Il; Furuzono, Tomoya [Laboratory of Molecular Function of Food, Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Uji 611-0011 (Japan); Takahashi, Nobuyuki [Laboratory of Molecular Function of Food, Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Uji 611-0011 (Japan); Research Unit for Physiological Chemistry, The Center for the Promotion of Interdisciplinary Education and Research, Kyoto University (Japan); Yamakuni, Kanae; Yang, Ha-Eun; Li, Yongjia [Laboratory of Molecular Function of Food, Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Uji 611-0011 (Japan); Ohue, Ryuji [Laboratory of Molecular Function of Food, Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Uji 611-0011 (Japan); Research Unit for Physiological Chemistry, The Center for the Promotion of Interdisciplinary Education and Research, Kyoto University (Japan); Nomura, Wataru [Laboratory of Molecular Function of Food, Division of Food Science and Biotechnology, Graduate School of Agriculture, Kyoto University, Uji 611-0011 (Japan); Sugawara, Tatsuya [Laboratory of Marine Bioproducts Technology, Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502 (Japan); Yu, Rina [Department of Food Science and Nutrition, University of Ulsan, Ulsan 680-749 (Korea, Republic of); Kitamura, Nahoko [Laboratory of Fermentation Physiology and Applied Microbiology, Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University, Kyoto 606-8502 (Japan); and others

    2015-04-17

    Our previous study has shown that gut lactic acid bacteria generate various kinds of fatty acids from polyunsaturated fatty acids such as linoleic acid (LA). In this study, we investigated the effects of LA and LA-derived fatty acids on the activation of peroxisome proliferator-activated receptors (PPARs) which regulate whole-body energy metabolism. None of the fatty acids activated PPARδ, whereas almost all activated PPARα in luciferase assays. Two fatty acids potently activated PPARγ, a master regulator of adipocyte differentiation, with 10-oxo-12(Z)-octadecenoic acid (KetoA) having the most potency. In 3T3-L1 cells, KetoA induced adipocyte differentiation via the activation of PPARγ, and increased adiponectin production and insulin-stimulated glucose uptake. These findings suggest that fatty acids, including KetoA, generated in gut by lactic acid bacteria may be involved in the regulation of host energy metabolism. - Highlights: • Most LA-derived fatty acids from gut lactic acid bacteria potently activated PPARα. • Among tested fatty acids, KetoA and KetoC significantly activated PPARγ. • KetoA induced adipocyte differentiation via the activation of PPARγ. • KetoA enhanced adiponectin production and glucose uptake during adipogenesis.

  4. 10-oxo-12(Z)-octadecenoic acid, a linoleic acid metabolite produced by gut lactic acid bacteria, potently activates PPARγ and stimulates adipogenesis

    Our previous study has shown that gut lactic acid bacteria generate various kinds of fatty acids from polyunsaturated fatty acids such as linoleic acid (LA). In this study, we investigated the effects of LA and LA-derived fatty acids on the activation of peroxisome proliferator-activated receptors (PPARs) which regulate whole-body energy metabolism. None of the fatty acids activated PPARδ, whereas almost all activated PPARα in luciferase assays. Two fatty acids potently activated PPARγ, a master regulator of adipocyte differentiation, with 10-oxo-12(Z)-octadecenoic acid (KetoA) having the most potency. In 3T3-L1 cells, KetoA induced adipocyte differentiation via the activation of PPARγ, and increased adiponectin production and insulin-stimulated glucose uptake. These findings suggest that fatty acids, including KetoA, generated in gut by lactic acid bacteria may be involved in the regulation of host energy metabolism. - Highlights: • Most LA-derived fatty acids from gut lactic acid bacteria potently activated PPARα. • Among tested fatty acids, KetoA and KetoC significantly activated PPARγ. • KetoA induced adipocyte differentiation via the activation of PPARγ. • KetoA enhanced adiponectin production and glucose uptake during adipogenesis

  5. Inflamm-aging and arachadonic acid metabolite differences with stage of tendon disease.

    Stephanie Georgina Dakin

    Full Text Available The contribution of inflammation to the pathogenesis of tendinopathy and high prevalence of re-injury is not well established, although recent evidence suggests involvement of prostaglandins. We investigated the roles of prostaglandins and inflammation-resolving mediators in naturally occurring equine tendon injury with disease stage and age. Levels of prostaglandins E(2 (PGE(2, F(2α (PGF(2α, lipoxin A(4 (LXA(4 and its receptor FPR2/ALX were analysed in extracts of normal, sub-acute and chronic injured tendons. To assess whether potential changes were associated with altered PGE(2 metabolism, microsomal prostaglandin E synthase-1 (mPGES-1, prostaglandin dehydrogenase (PGDH, COX-2 and EP(4 receptor expression were investigated. The ability of tendons to resolve inflammation was determined by assessing FPR2/ALX expression in natural injury and IL-1β stimulated tendon explants.Alterations in the profile of lipid mediators during sub-acute injury included low PGE(2 and elevated LXA(4 levels compared to normal and chronic injuries. In contrast, PGF(2α levels remained unchanged and were three-fold lower than PGE(2. The synthetic capacity of PGE(2 as measured by the ratio of mPGES-1:PGDH was elevated in sub-acute injury, suggesting aberrations in tendon prostaglandin metabolism, whilst COX-2 and EP(4 receptor were unchanged. Paradoxically low tendon PGE(2 levels in early injury may be attributed to increased local clearance via PGDH or the class switching of lipid mediators from the prostaglandin to the lipoxin axis. PGE(2 is therefore implicated in the development of tendon inflammation and its ensuing resolution. Whilst there was no relationship between age and tendon LXA(4 levels, there was an age-associated decline in FPR2/ALX receptor expression with concurrent increased PGE(2 levels in injury. Furthermore, uninjured tendon explants from younger (<10 years but not older horses (≥10 years treated with IL-1β responded by increasing FPR2/ALX

  6. Two Sets of Paralogous Genes Encode the Enzymes Involved in the Early Stages of Clavulanic Acid and Clavam Metabolite Biosynthesis in Streptomyces clavuligerus

    Tahlan, Kapil; Park, Hyeon Ung; Wong, Annie; Beatty, Perrin H.; Jensen, Susan E.

    2004-01-01

    Recently, a second copy of a gene encoding proclavaminate amidinohydrolase (pah1), an enzyme involved in the early stages of clavulanic acid and clavam metabolite biosynthesis in Streptomyces clavuligerus, was identified and isolated. Using Southern analysis, we have now isolated second copies of the genes encoding the carboxyethylarginine synthase (ceaS) and β-lactam synthetase (bls) enzymes. These new paralogues are given the gene designations ceaS1 and bls1 and are located immediately upst...

  7. Relative developmental toxicity of glycol ether alkoxy acid metabolites in the embryonic stem cell test as compared with the in vivo potency of their parent compounds.

    de Jong, Esther; Louisse, Jochem; Verwei, Miriam; Blaauboer, Bas J; van de Sandt, Johannes J M; Woutersen, Ruud A; Rietjens, Ivonne M C M; Piersma, Aldert H

    2009-07-01

    The embryonic stem cell test (EST) has been proposed as an in vitro assay that might reduce animal experimentation in regulatory developmental toxicology. So far, evaluation of the EST was not performed using compounds within distinct chemical classes. Evaluation within a distinct class of chemically related compounds can define the usefulness of the assay for the chemical class tested. The aim of the present study was to evaluate the relative sensitivity of the EST for a selected series of homologous compounds and to compare the data to the relative developmental toxicity of the compounds in vivo. To this end a series of proximate developmentally toxic glycol ether alkoxy acid metabolites was tested in the EST. All glycol ether alkoxy acid metabolites tested showed a concentration-dependent inhibition of cardiomyocyte differentiation at noncytotoxic concentrations, with methoxyacetic acid as the most potent compound followed by ethoxyacetic acid, butoxyacetic acid, and phenoxyacetic acid, respectively. The potency ranking of the compounds in the EST corresponds with the available in vivo data. The relative differences between the potencies of the compounds appeared more pronounced in the in vivo studies than in the EST. A possible explanation for this discrepancy could be the difference in the kinetics of the compounds in vivo as compared with their in vitro kinetics. This study illustrates that the EST can be used to set priorities for developmental toxicity testing within classes of related compounds. PMID:19401353

  8. Investigations on some metabolites of Tecoma stans Juss. callus tissue. Part III. Chromatographical search for iridoids, phenolic acids, terpenoids and sugars

    Barbara Dohnal

    2015-05-01

    Full Text Available Tissus cultures of Tecoma stans Juss. cultivated on modified Murashige-Skoog medium (RT-k were phytochemically analysed by means of chromatographical methods (PC, TLC. The following products were found as metabolites: phenolic acids - chlorogenics, caffeic, ferulic, vanillic, o-coumaric and sinapic; steroids - β-sitosterol; triterpenes - ursolic and oleanolic acids, α-amyrine; sugars - glucose, fructose, sucrose, xylose. Meso-inositol was isolated in 0.8% yield. In intact plant leaves, some differences concerning the content and/or number of individual compounds were observed, namely: lack of sinapic acid and occurrence of p-coumaric acid, lower content of β-sitosterol, lack of oleanolic acid, occurrence of β-amyrine and of one unidentified triterpenoid, lack of xylose, occurrence of maltose, raffinose, and stachiose. The level of mesoinositol inn leaves was distincly lower than in the callus tissues. Neither in callus tissues nor in leaves iridoid glycosides were found.

  9. Contribution to the study of the behaviour of the diethylene triamine pentaacetic acid in acid medium

    After an overview of the evolutions and developments brought to reprocessing methods and techniques for the decontamination and separation of nuclear wastes, the author reports the study of the behaviour of a complexing compound (DTPA, diethylene triamine pentaacetic acid) in acid medium, and of the complexing with plutonium 239. After a presentation of DTPA properties, the author reports the study of its behaviour in acid medium. The next part reports the study of its reaction possibilities in acid medium: Th(IV)-DTPA complexing, Pu(IV)-DTPA complexing. Different techniques have been used: infrared spectrometry, potentiometry, spectrophotometry, molar rate method. The author then describes the application of DTPA properties in acid medium to the processing of irradiated targets

  10. A liquid chromatography/mass spectrometric method for simultaneous analysis of arachidonic acid and its endogenous eicosanoid metabolites prostaglandins, dihydroxyeicosatrienoic acids, hydroxyeicosatetraenoic acids, and epoxyeicosatrienoic acids in rat brain tissue.

    Yue, Hongfei; Jansen, Susan A; Strauss, Kenneth I; Borenstein, Michael R; Barbe, Mary F; Rossi, Luella J; Murphy, Elise

    2007-02-19

    A sensitive, specific, and robust liquid chromatography/mass spectrometric (LC/MS) method was developed and validated that allows simultaneous analysis of arachidonic acid (AA) and its cyclooxygenase, cytochrome P450, and lipoxygenase pathway metabolites prostaglandins (PGs), dihydroxyeicosatrienoic acids (DiHETrEs), hydroxyeicosatetraenoic acids (HETEs) and epoxyeicosatrienoic acids (EETs), including PGF(2alpha), PGE(2), PGD(2), PGJ(2), 14,15-DiHETrE, 11,12-DiHETrE, 8,9-DiHETrE, 5,6-DiHETrE, 20-HETE, 15-HETE, 12-HETE, 9-HETE, 8-HETE, 5-HETE, 14,15-EET, 11,12-EET, 8,9-EET, and 5,6-EET in rat brain tissues. Deuterium labeled PGF(2alpha)-d(4), PGD(2)-d(4), 15(S)-HETE-d(8), 14,15-EET-d(8), 11,12-EET-d(8), 8,9-EET-d(8), and AA-d(8) were used as internal standards. Solid phase extraction was used for sample preparation. A gradient LC/MS method using a C18 column and electrospray ionization source under negative ion mode was optimized for the best sensitivity and separation within 35 min. The method validation, including LC/MS instrument qualification, specificity, calibration model, accuracy, precision (without brain matrix and with brain matrix), and extraction efficiency were performed. The linear ranges of the calibration curves were 2-1000 pg for PGs, DiHETrEs, HETEs, and EETs, 10-2400 pg for PGE(2) and PGD(2), and 20-2000 ng for AA, respectively. PMID:17125954

  11. Liquid chromatography–tandem mass spectrometry method for simultaneous determination of valproic acid and its ene-metabolites in epilepsy patient plasma

    Huan Lu

    2016-04-01

    Full Text Available A simple and high throughput method was developed and validated for simultaneous determination of valproic acid and its two toxicant ene-metabolites, 2-enevalproic acid and 4-enevalproic acid in epilepsy patient plasma using liquid chromatography–tandem mass spectrometry. Probenecid was used as internal standard and solid-phase extraction was selected for sample preparation. A chromatographic separation was performed on an Agilent Poroshell SB-C18 column (50 mm×4.6 mm i.d., 2.7 μm by an optimized gradient elution at a flow rate of 0.9 mL/min. The total run time was 7 min. Electrospray ionization was used in negative ion mode by multiple reaction monitoring of the precursor-to-product ion transitions at m/z 143.0→143.0 for valproic acid, m/z 140.9→140.9 for 2-enevalproic acid and 4-enevalproic acid for their poor fragments, and m/z 283.9→239.9 for probenecid. The results showed good linearity of valproic acid, 2-enevalproic acid and 4-enevalproic acid in their respective linear ranges. The correlation coefficients were more than 0.998. The intra- and inter-day precision of the assay was less than 11.0% and the accuracy ranged from 2% to 12%. This analytical method was successfully applied to assay plasma concentrations of valproic acid and its two ene-metabolites in epilepsy patient plasma and used for therapeutic drug monitoring.

  12. Organic Anion Transporting Polypeptide (OATP)2B1 Contributes to Gastrointestinal Toxicity of Anticancer Drug SN-38, Active Metabolite of Irinotecan Hydrochloride.

    Fujita, Daichi; Saito, Yoshimasa; Nakanishi, Takeo; Tamai, Ikumi

    2016-01-01

    Gastrointestinal toxicity, such as late-onset diarrhea, is a significant concern in irinotecan hydrochloride (CPT-11)-containing regimens. Prophylaxis of late-onset diarrhea has been reported with use of Japanese traditional (Kampo) medicine containing baicalin and with the antibiotic cefixime, and this has been explained in terms of inhibition of bacterial deconjugation of SN-38-glucuronide since unconjugated SN-38 (active metabolite of CPT-11) is responsible for the gastrointestinal toxicity. It is also prerequisite for SN-38 to be accumulated in intestinal tissues to exert toxicity. Based on the fact that liver-specific organic anion transporting polypeptide (OATP)1B1, a member of the same family as OATP2B1, is known to be involved in hepatic transport of SN-38, we hypothesized that intestinal transporter OATP2B1 contributes to the accumulation of SN-38 in gastrointestinal tissues, and its inhibition would help prevent associated toxicity. We found that uptake of SN-38 by OATP2B1-expressing Xenopus oocytes was significantly higher than that by control oocytes. OATP2B1-mediated uptake of SN-38 was saturable, pH dependent, and decreased in the presence of baicalin, cefixime, or fruit juices such as apple juice. In vivo gastrointestinal toxicity of SN-38 in mice caused by oral administration for consecutive 5 days was prevented by coingestion of apple juice. Thus, OATP2B1 contributes to the uptake of SN-38 by intestinal tissues, triggering gastrointestinal toxicity. So, in addition to the reported inhibition of bacterial β-glucuronidase by cefixime or baicalin, inhibition of OATP2B1 may also contribute to prevention of gastrointestinal toxicity. Apple juice may be helpful for prophylaxis of late-onset diarrhea observed in CPT-11 therapy without disturbance of the intestinal microflora. PMID:26526067

  13. Impact of different plant secondary metabolites addition: saponin, tannic acid, salicin and aloin on glucose anaerobic co-digestion

    Mambanzulua Ngoma, Philippe; Hiligsmann, Serge; Sumbu Zola, Eric; Ongena, Marc; Thonart, Philippe

    2015-01-01

    Vegetal waste and some wastewater of agro-food industries contain plant secondary metabolites (PSMs). It was showed in nutritional researches that these substances such as saponins and tannins reduced the methane production in the rumen. To our knowledge no study was done in the waste treatment domain to evaluate the inhibitory effect of the principal glycosidic metabolites from the wastewater or vegetal waste on their own methane-producing anaerobic digestion. Therefore in this paper BMP ...

  14. Enhanced metabolite generation

    Chidambaram, Devicharan

    2012-03-27

    The present invention relates to the enhanced production of metabolites by a process whereby a carbon source is oxidized with a fermentative microbe in a compartment having a portal. An electron acceptor is added to the compartment to assist the microbe in the removal of excess electrons. The electron acceptor accepts electrons from the microbe after oxidation of the carbon source. Other transfers of electrons can take place to enhance the production of the metabolite, such as acids, biofuels or brewed beverages.

  15. Sequential determination of metabolites involved in the biosynthesis of aromatic amino acids after ultrasound-assisted extraction from plants and reverse LC separation.

    Alcaide-Molina, Miguel; Priego-Capote, Feliciano; Luque de Castro, María Dolores

    2013-02-15

    A dual method is proposed for the determination of metabolites involved in the shikimate pathway which are biomarkers of the effects of glyphosate action on plants exposed to this herbicide. Extraction of the target metabolites (phenylalanine, tryptophan, tyrosine and shikimic acid) from a wheat model plant was accelerated by ultrasound energy. After centrifugation and micro-filtration, 1 μL of extract was injected into the chromatograph in an isocratic regime for 4 min to determine shikimate by absorption at 254 nm. In the mean time, a 130 μL aliquot of extract was subjected to derivatization with o-phthaldialdehyde and 2-mercaptoethanol for 1 min, the reaction stopped and 1 μL of the solution chromatographied in a gradient regime prior to laser-induced fluorescence detection of the derivatized amino acids. The characterization of the dual method provided limits of detection around 0.03 μg mL(-1) for the aromatic amino acids and 1.52 μg mL(-1) for shikimate, whereas the limits of quantitation ranged between 0.084 and 0.093 μg mL(-1) for amino acids and was of 4.56 μg mL(-1) for shikimate. The suitability of the method was checked by application to Triticum aestivum (wheat) plants grown under controlled conditions, sprayed with different doses of glyphosate and collected at different times after exposition to the herbicide. PMID:23598041

  16. Increased formic acid excretion and the development of kidney toxicity in rats following chronic dosing with trichloroethanol, a major metabolite of trichloroethylene

    The chronic toxicity of trichloroethanol, a major metabolite of trichloroethylene, has been assessed in male Fischer rats (60 per group) given trichloroethanol in drinking water at concentrations of 0, 0.5 and 1.0 g/l for 52 weeks. The rats excreted large amounts of formic acid in urine reaching a maximum after 12 weeks (∼65 mg/24 h at 1 g/l) and thereafter declining to reach an apparent steady state at 40 weeks (15-20 mg/24 h). Urine from treated rats was more acidic throughout the study and urinary methylmalonic acid and plasma N-methyltetrahydrofolate concentrations were increased, indicating an acidosis, vitamin B12 deficiency and impaired folate metabolism, respectively. The rats treated with trichloroethanol developed kidney damage over the duration of the study which was characterised by increased urinary NAG activity, protein excretion (from 4 weeks), increased basophilia, protein accumulation and tubular damage (from 12 to 40 weeks), increased cell replication (at week 28) and evidence in some rats of focal proliferation of abnormal tubules at 52 weeks. It was concluded that trichloroethanol, the major metabolite of trichloroethylene, induced nephrotoxicity in rats as a result of formic acid excretion and acidosis

  17. Multivariate data analysis for finding the relevant fatty acids contributing to the melting fractions of cream

    Buldo, Patrizia; Larsen, Mette Krogh; Wiking, Lars

    2013-01-01

    palmitoleic acid (C16:1) in milk fat, whereas it decreased the amount of stearic acid (C18:0) and C18:1 trans fatty acid. Average data on the melting behaviour of cream separated the farms into two groups where the main differences in feeding were the amounts of maize silage and rapeseed cake used. CONCLUSION......BACKGROUND: The melting behaviour and fatty acid composition of cream from a total of 33 cows from four farms were analysed. Multivariate data analysis was used to identify the fatty acids that contributed most to the melting points and to differentiate between creams from different practical...... feeding regimes. RESULTS: It was demonstrated that the melting point of the medium melting fraction of milk fat was positively correlated with palmitic acid (C16:0), whereas it was negatively correlated with oleic acid (C18:1 cis9), conjugated linoleic acid (CLA cis9 trans11), vaccenic acid (C18:1 trans11...

  18. The effects of dietary omega fatty acids on pregnancy rate, plasma prostaglandin metabolite levels, serum progesterone levels, and milk fatty-acid profile in beef cows.

    Richardson, Gavin F; McNiven, Mary A; Petit, Hélène V; Duynisveld, John L

    2013-10-01

    The objectives were to determine the effects of feeding supplements rich in omega-6 or omega-3 fatty acids (FA) during the late gestation to the early postpartum and breeding periods on reproduction and milk FA profile in beef cows. For each of two years, at the beginning of period 1 (mid-December), 72 beef cows, calving in January or February, were assigned to diets supplemented with roasted flaxseed (Flax) or roasted soybean (Soybean). For each of two years, after 11 wk (end of period 1), 18 cows of 36 in the Flax group were switched to the soybean supplement and 18 cows of 36 in the Soybean group were switched to the flax supplement (start of Period 2). Cows were bred by timed artificial insemination (TAI) in week 5 of period 2. The FA composition of the milk reflected the FA profile of the oilseed supplements. There were no differences in pregnancy rates among the 4 groups. The treatments had no effect on plasma prostaglandin metabolite levels or ratios at 4 to 11 d postpartum. At 5 to 6 d post- TAI, pregnant cows fed Flax in period 1 had lower (P Flax in period 2 had higher (P < 0.05) serum progesterone levels at 5 to 6 d post-TAI than cows fed Soybean, but there was no difference at 19 to 20 d post-TAI. The dietary treatments had no effect on pregnancy rates, but there were some effects on plasma PGFM levels, PGFM to PGEM ratios, and serum progesterone levels. The FA supplements influenced the FA composition of milk. PMID:24124276

  19. Reevaluating the contribution of sulfuric acid and the origin of organic compounds in atmospheric nanoparticle growth

    Vakkari, Ville; Tiitta, Petri; Jaars, Kerneels; Croteau, Philip; Beukes, Johan Paul; Josipovic, Miroslav; Kerminen, Veli-Matti; Kulmala, Markku; Venter, Andrew D.; Zyl, Pieter G.; Worsnop, Douglas R.; Laakso, Lauri

    2015-12-01

    Aerosol particles formed in the atmosphere are important to the Earth's climate system due to their ability to affect cloud properties. At present, little is known about the atmospheric chemistry responsible for the growth of newly formed aerosol particles to climate-relevant sizes. Here combining detailed aerosol measurements with a theoretical framework we found that depending on the gaseous precursors and size of the newly formed particles, the growth was dominated by either sulfuric acid accompanied by ammonium or organic compounds originating in either biogenic emissions or savannah fires. The contribution of sulfuric acid was larger during the early phases of the growth, but in clean conditions organic compounds dominated the growth from 1.5 nm up to climatically relevant sizes. Furthermore, our analysis indicates that in polluted environments the contribution of sulfuric acid to the growth may have been underestimated by up to a factor of 10.

  20. Np(V) reduction by humic acid: Contribution of reduced sulfur functionalities to the redox behavior of humic acid

    The role of sulfur-containing functional groups in humic acids for the Np(V) reduction in aqueous solution has been studied with the objective to specify individual processes contributing to the overall redox activity of humic substances. For this, humic acid model substances type M1-S containing different amounts of sulfur (1.9, 3.9, 6.9 wt.%) were applied. The sulfur functionalities in these humic acids are dominated by reduced-sulfur species, such as thiols, dialkylsulfides and/or disulfides. The Np(V) reduction behavior of these humic acids has been studied in comparison to that of the sulfur-free humic acid type M1 at pH 5.0, 7.0 and 9.0 under anaerobic conditions by means of batch experiments. For Np redox speciation in solution, solvent extraction and ultrafiltration were applied. In addition, redox potentials of the sample solutions were monitored. At pH 5.0, both rate and extent of Np(V) to Np(IV) reduction were found to increase with increasing sulfur content of the humic acids. At pH 7.0 and 9.0, sulfur functional groups had only a slight influence on the reduction behavior of humic acid toward Np(V). Thus, in addition to quinoid moieties and non-quinoid phenolic OH groups, generally acknowledged as main redox-active sites in humic substances, sulfur functional groups have been identified as further redox-active moieties of humic substances being active especially in the slightly acidic pH range as shown for Np(V). Due to the low sulfur content of up to 2 wt.% in natural humic substances, their contribution to the total reducing capacity is smaller than that of the other redox-active functional groups. - Highlights: ► Redox processes of humic substances control the speciation and mobility of metal ions in the environment. ► Sulfur functional groups were identified as further redox-active sites in humic substances toward Np(V). ► The specification of processes contributing to the redox capacity of humics improves the prediction of actinide migration.

  1. Growth inhibitory effect of polyunsaturated fatty acids (PUFAs on colon cancer cells via their growth inhibitory metabolites and fatty acid composition changes.

    Chengcheng Zhang

    Full Text Available Colorectal cancer is common. Polyunsaturated fatty acids (PUFAs exert growth-inhibitory and pro-apoptotic effects on colon cancer cells. Metabolites of PUFAs such as prostaglandins (PGs, leukotrienes (LTs and lipoxins (LXs play a significant role in colon cancer.Human colon cancer LoVo and RKO cells were cultured with different concentration of PUFAs and 5-fluorouracil (5-FU in vitro. Cell morphological changes, fatty acid composition, formation of PGE2, LTB4 and LXA4 and expression of COX-2, ALOX5, PGD synthase (PGDS, microsomal prostaglandin E synthase (mPGES were assessed in LoVo and RKO cells when supplemented with PUFAs and 5-FU.PUFAs and 5-FU inhibited growth of LoVo and RKO cells to the same extent at the doses used and produced significant alterations in their shape. As expected, higher concentrations of supplemented PUFAs were noted in the cells compared to control. LA, GLA, AA, ALA and EPA supplementation to LoVo cells suppressed production of PGE2, LTB4,and ALOX5, mPGES expression, but enhanced that of LXA4; whereas DHA enhanced PGE2 and LXA4 synthesis but decreased LTB4 formation and COX-2, ALOX5, mPGES expression. In contrast, 5-FU enhanced formation of PGE2, LTB4 and mPGES expression, but suppressed LXA4 synthesis and COX-2 expression. PGE2, LTB4 synthesis and ALOX5 expression was suppressed by LA, GLA, ALA and DHA; whereas AA, EPA and 5-FU enhanced PGE2 but paradoxically AA decreased and EPA and 5-FU enhanced LTB4 synthesis in RKO cells. All the PUFAs tested enhanced, while 5-FU decreased LXA4 formation in RKO cells; whereas GLA, AA, and 5-FU augmented while LA, ALA, EPA and DHA enhanced COX-2 expression in RKO cells.Tumoricidal action of PUFAs on colorectal LoVo and RKO cancer cells in vitro was associated with increased formation of LXA4, decreased synthesis of PGE2 and LTB4 and suppressed expression of COX-2, ALOX5, mPGES, whereas 5-FU produced contrasting actions on these indices.

  2. Contribution of sulfuric acid and oxidized organic compounds to particle formation and growth

    F. Riccobono

    2012-10-01

    Full Text Available Lack of knowledge about the mechanisms underlying new particle formation and their subsequent growth is one of the main causes for the large uncertainty in estimating the radiative forcing of atmospheric aerosols in global models. We performed chamber experiments designed to study the contributions of sulfuric acid and organic vapors to the formation and early growth of nucleated particles. Distinct experiments in the presence of two different organic precursors (1,3,5-trimethylbenzene and α-pinene showed the ability of these compounds to reproduce the formation rates observed in the low troposphere. These results were obtained measuring the sulfuric acid concentrations with two chemical ionization mass spectrometers confirming the results of a previous study which modeled the sulfuric acid concentrations in presence of 1,3,5-trimethylbenzene.

    New analysis methods were applied to the data collected with a condensation particle counter battery and a scanning mobility particle sizer, allowing the assessment of the size resolved growth rates of freshly nucleated particles. The effect of organic vapors on particle growth was investigated by means of the growth rate enhancement factor (Γ, defined as the ratio between the measured growth rate in the presence of α-pinene and the kinetically limited growth rate of the sulfuric acid and water system. The observed Γ values indicate that the growth is already dominated by organic compounds at particle diameters of 2 nm. Both the absolute growth rates and Γ showed a strong dependence on particle size, supporting the nano-Köhler theory. Moreover, the separation of the contributions from sulfuric acid and organic compounds to particle growth reveals that the organic contribution seems to be enhanced by the sulfuric acid concentration. Finally, the size resolved growth analysis indicates that both condensation of oxidized organic compounds and reactive uptake contribute to particle growth.

  3. Contribution of sulfuric acid and oxidized organic compounds to particle formation and growth

    F. Riccobono

    2012-05-01

    Full Text Available Lack of knowledge about the mechanisms underlying new particle formation and their subsequent growth is one of the main causes for the large uncertainty in estimating the radiative forcing of atmospheric aerosols in global models. We performed chamber experiments designed to study the contributions of sulfuric acid and organic vapors to formation and to the early growth of nucleated particles, respectively. Distinct experiments in the presence of two different organic precursors (1,3,5-trimethylbenzene and α-pinene showed the ability of these compounds to reproduce the formation rates observed in the low troposphere. These results were obtained measuring the sulfuric acid concentrations with two Chemical Ionization Mass Spectrometers confirming the results of a previous study which modeled the sulfuric acid concentrations in presence of 1,3,5-trimethylbenzene.

    New analysis methods were applied to the data collected with a Condensation Particle Counter battery and a Scanning Mobility Particle Sizer, allowing the assessment of the size resolved growth rates of freshly nucleated particles. The effect of organic vapors on particle growth was investigated by means of the growth rate enhancement factor (Γ, defined as the ratio between the measured growth rate in the presence of α-pinene and the kinetically limited growth rate of the sulfuric acid and water system. The observed Γ values indicate that the growth is dominated by organic compounds already at particle diameters of 2 nm. Both the absolute growth rates and Γ showed a strong dependence on particle size supporting the nano-Köhler theory. Moreover, the separation of the contributions from sulfuric acid and organic compounds to particles growth reveals that the organic contribution seems to be enhanced by the sulfuric acid concentration. The size resolved growth analysis finally indicates that both condensation of oxidized organic compounds and reactive uptake contribute to

  4. Human urine certified reference material: Creatinine and toluene metabolites: O-cresol and hippuric acid (preliminary studies)

    Šperlingová, I.; Dabrowská, L.; Stránský, V.; Kučera, Jan; Tichý, M.

    2005-01-01

    Roč. 355, - (2005), s. 411-411. ISSN 0009-8981 Institutional research plan: CEZ:AV0Z10480505 Keywords : certified reference material * human urine * , creatinine and toluene metabolites Subject RIV: BG - Nuclear, Atomic and Molecular Physics, Colliders Impact factor: 2.149, year: 2005

  5. Cyclohexane-1,2-dicarboxylic acid diisononyl ester and metabolite effects on rat epididymal stromal vascular fraction differentiation of adipose tissue

    Plastics are generally mixed with additives like plasticizers to enhance their flexibility, pliability, and elasticity proprieties. Plasticizers are easily released into the environment and are absorbed mainly through ingestion, dermal contact, and inhalation. One of the main classes of plasticizers, phthalates, has been associated with endocrine and reproductive diseases. In 2002, 1,2-cyclohexane dicarboxylic acid diisononyl ester (DINCH) was introduced in the market for use in plastic materials and articles intended to come into contact with food, and it received final approval from the European Food Safety Authority in 2006. At present, there is limited knowledge about the safety and potential metabolic and endocrine-disrupting properties of DINCH and its metabolites. The purpose of this study was to evaluate the biological effects of DINCH and its active metabolites, cyclohexane-1,2-dicarboxylic acid (CHDA) and cyclohexane-1,2-dicarboxylic acid mono isononyl ester (MINCH), on rat primary stromal vascular fraction (SVF) of adipose tissue. DINCH and its metabolite, CHDA, were not able to directly affect SVF differentiation. However, exposure of SVF to 50 μM and 100 μM concentrations of MINCH affected the expression of Cebpa and Fabp4, thus inducing SVF preadipocytes to accumulate lipids and fully differentiate into mature adipocytes. The effect of MINCH was blocked by the specific peroxisome proliferator-activated receptor (PPAR)-α antagonist, GW6471. Taken together, these results suggest that MINCH is a potent PPAR-α agonist and a metabolic disruptor, capable of inducing SVF preadipocyte differentiation, that may interfere with the endocrine system in mammals. - Highlights: • DINCH and CHDA did not affect the adipogenesis of the SVF. • MINCH affected the adipogenesis of the SVF. • MINCH effect was blocked by the specific PPAR-α antagonist GW6471. • MINCH exerted a similar effect as MEHP on SVF adipogenesis. • DINCH/MINCH are potential metabolic

  6. Cyclohexane-1,2-dicarboxylic acid diisononyl ester and metabolite effects on rat epididymal stromal vascular fraction differentiation of adipose tissue

    Campioli, Enrico [Research Institute of the McGill University Health Centre (Canada); Department of Medicine, McGill University, Montréal, Québec (Canada); Duong, Tam B. [Research Institute of the McGill University Health Centre (Canada); Deschamps, François [Synthèse AptoChem Inc., Montréal, Québec (Canada); Papadopoulos, Vassilios, E-mail: vassilios.papadopoulos@mcgill.ca [Research Institute of the McGill University Health Centre (Canada); Department of Medicine, McGill University, Montréal, Québec (Canada); Department of Biochemistry, McGill University, Montréal, Québec (Canada); Department of Pharmacology and Therapeutics, McGill University, Montréal, Québec (Canada)

    2015-07-15

    Plastics are generally mixed with additives like plasticizers to enhance their flexibility, pliability, and elasticity proprieties. Plasticizers are easily released into the environment and are absorbed mainly through ingestion, dermal contact, and inhalation. One of the main classes of plasticizers, phthalates, has been associated with endocrine and reproductive diseases. In 2002, 1,2-cyclohexane dicarboxylic acid diisononyl ester (DINCH) was introduced in the market for use in plastic materials and articles intended to come into contact with food, and it received final approval from the European Food Safety Authority in 2006. At present, there is limited knowledge about the safety and potential metabolic and endocrine-disrupting properties of DINCH and its metabolites. The purpose of this study was to evaluate the biological effects of DINCH and its active metabolites, cyclohexane-1,2-dicarboxylic acid (CHDA) and cyclohexane-1,2-dicarboxylic acid mono isononyl ester (MINCH), on rat primary stromal vascular fraction (SVF) of adipose tissue. DINCH and its metabolite, CHDA, were not able to directly affect SVF differentiation. However, exposure of SVF to 50 μM and 100 μM concentrations of MINCH affected the expression of Cebpa and Fabp4, thus inducing SVF preadipocytes to accumulate lipids and fully differentiate into mature adipocytes. The effect of MINCH was blocked by the specific peroxisome proliferator-activated receptor (PPAR)-α antagonist, GW6471. Taken together, these results suggest that MINCH is a potent PPAR-α agonist and a metabolic disruptor, capable of inducing SVF preadipocyte differentiation, that may interfere with the endocrine system in mammals. - Highlights: • DINCH and CHDA did not affect the adipogenesis of the SVF. • MINCH affected the adipogenesis of the SVF. • MINCH effect was blocked by the specific PPAR-α antagonist GW6471. • MINCH exerted a similar effect as MEHP on SVF adipogenesis. • DINCH/MINCH are potential metabolic

  7. The gamma-aminobutyric acid shunt contributes to closing the tricarboxylic acid cycle in Synechocystis sp PCC 6803

    Xiong, W; Brune, D; Vermaas, WFJ

    2014-07-16

    A traditional 2-oxoglutarate dehydrogenase complex is missing in the cyanobacterial tricarboxylic acid cycle. To determine pathways that convert 2-oxoglutarate into succinate in the cyanobacterium Synechocystis sp. PCC 6803, a series of mutant strains, Delta sll1981, Delta slr0370, Delta slr1022 and combinations thereof, deficient in 2-oxoglutarate decarboxylase (Sll1981), succinate semialdehyde dehydrogenase (Slr0370), and/or in gamma-aminobutyrate metabolism (Slr1022) were constructed. Like in Pseudomonas aeruginosa, N-acetylornithine aminotransferase, encoded by slr1022, was shown to also function as gamma-aminobutyrate aminotransferase, catalysing gamma-aminobutyrate conversion to succinic semialdehyde. As succinic semialdehyde dehydrogenase converts succinic semialdehyde to succinate, an intact gamma-aminobutyrate shunt is present in Synechocystis. The Delta sll1981 strain, lacking 2-oxoglutarate decarboxylase, exhibited a succinate level that was 60% of that in wild type. However, the succinate level in the Delta slr1022 and Delta slr0370 strains and the Delta sll1981/Delta slr1022 and Delta sll1981/Delta slr0370 double mutants was reduced to 20-40% of that in wild type, suggesting that the gamma-aminobutyrate shunt has a larger impact on metabolite flux to succinate than the pathway via 2-oxoglutarate decarboxylase. C-13-stable isotope analysis indicated that the gamma-aminobutyrate shunt catalysed conversion of glutamate to succinate. Independent of the 2-oxoglutarate decarboxylase bypass, the gamma-aminobutyrate shunt is a major contributor to flux from 2-oxoglutarate and glutamate to succinate in Synechocystis sp. PCC 6803.

  8. Comparative bioavailability of two oral formulations of clopidogrel: Determination of clopidogrel and its carboxylic acid metabolite (SR26334 under fasting and fed conditions in healthy subjects

    Brvar Nina

    2014-03-01

    Full Text Available Two randomized, single dose, 2-period, 2-sequence crossover studies were conducted to evaluate the comparative bioavailability of two clopidogrel formulations under fasting and fed conditions. Assessment of bioequivalence was based upon measurement of plasma concentrations of the parent drug, clopidogrel, and its major (inactive metabolite, clopidogrel carboxylic acid, using improved methanol-free extraction. Bioequivalence of Krka’s formulation to the innovator’s formulation was demonstrated under both fasting and fed conditions on 205 volunteers. Confidence intervals for AUC0-t, AUC0-inf and Cmax of clopidogrel and its main metabolite were well within the acceptance range of 80.00 to 125.00 %. Food substantially increased the bioavailability of clopidogrel from both formulations, while no effect of food on the extent and rate of exposure to the metabolite was observed. The effect of food was comparable between the two formulations, as indicated by the same direction and rank of food impact on the bioavailability of both formulations.

  9. Characterization of in vitro and in vivo metabolites of carnosic acid, a natural antioxidant, by high performance liquid chromatography coupled with tandem mass spectrometry.

    Song, Yuelin; Yan, Haixia; Chen, Jinfeng; Wang, Yitao; Jiang, Yong; Tu, Pengfei

    2014-02-01

    Carnosic acid (CA) is a widely employed antioxidant and the main active component in rosemary and sage, but its metabolism remains largely unknown. The present study investigated the metabolism of CA in vitro and in vivo for the first time, using high performance liquid chromatography coupled with hybrid triple quadrupole-linear ion trap mass spectrometry (HPLC-Q-trap-MS). A couple of scan modes were adopted in mass spectrometer domain, including Q1 full scan, neutral loss scan-information dependent acquisition-enhanced product ion (NL-IDA-EPI) and precursor ion scan-information dependent acquisition-enhanced product ion (PI-IDA-EPI). In particular, a prediction was carried out on the basis of in vitro metabolism results, and gave birth to a multiple ion monitoring-information dependent acquisition-enhanced product ion (MIM-IDA-EPI) mode aiming to detect the trace metabolites in CA-treated biological samples. A total of ten metabolites (M4-13), along with three degradative products (M1-3), were identified for CA from in vitro metabolism models, including liver microsomes of human and rats (HLMs and RLMs), human intestinal microsomes (HIMs) and two species of Cunninghamella elegans. Twelve (U1-12) and six (F1-6) metabolites were detected from CA-treated urine and feces, respectively. In addition, five metabolites (SM2-6) in vivo were purified and definitely identified using NMR spectroscopy. The results of both in vitro and in vivo metabolism studies indicated poor metabolic stability for CA, and the glucuronidation and oxidation metabolisms extensively occurred for CA in vitro, while oxidation, glucuronidation and methylation were the main metabolic pathways observed in vivo. PMID:24291799

  10. Immunoregulation of antitumor response; differential secretion of arachidonic acid metabolites by macrophages during stimulation ''in vitro'' with BCG and ''Corynebacterium parvum''

    The level of arachidonic acid (AA) metabolites in the supernatants of cultures peritoneal exudate cells (PEC) were studied under various conditions using BCG and ''Corynebacterium parvum'' as stimulators. The metabolite levels were analyzed by thin layer chromatography (TLC). The degree of macrophage cytotoxic/cytostatic activity was dependent on the dose and character of stimulators used and the source of macrophages. The application of micro cytotoxicity assay for the evaluation of tumor cell lysis (lung sarcoma SaL-1) ''in vitro'' revealed that peritoneal macrophages from healthy and tumor bearing BALB/c mice may affect the degree of antitumor response. In the supernatants of cultured PEC from tumor bearing mice AA level increased (by 10-fold) in comparison with PEC from healthy mice. Stimulation with BCG induced over a double level of AA in PEC isolated from tumor bearing mice non-stimulated or stimulated with ''C.parvum''. A lower level of prostaglandins (PGs) was found in the supernatants of cultured PEC isolated from healthy mice (stimulated and non-stimulated), but the highest level of PGs was observed in the supernatants of cultured PEC isolated from tumor bearing mice stimulated with BCG. The unique metabolite of AA was found only in the supernatants form non-stimulated PEC from tumor bearing mice. PEC from tumor bearing mice produced metabolites of AA which were not detected in control group. These results suggest that macrophages also play a regulatory role by secretion of AA. This process can be modified by bacterial antigens. (author). 21 refs, 7 figs

  11. The contribution of mediated oxidation mechanisms in the electrolytic degradation of cyanuric acid using diamond anodes.

    Bensalah, Nasr; Dbira, Sondos; Bedoui, Ahmed

    2016-07-01

    In this work, the contribution of mediated oxidation mechanisms in the electrolytic degradation of cyanuric acid using boron-doped diamond (BDD) anodes was investigated in different electrolytes. A complete mineralization of cyanuric acid was obtained in NaCl; however lower degrees of mineralization of 70% and 40% were obtained in Na2SO4 and NaClO4, respectively. This can be explained by the nature of the oxidants electrogenerated in each electrolyte. It is clear that the contribution of active chlorine (Cl2, HClO, ClO(-)) electrogenerated from oxidation of chlorides on BDD is much more important in the electrolytic degradation of cyanuric acid than the persulfate and hydroxyl radicals produced by electro-oxidation of sulfate and water on BDD anodes. This could be explained by the high affinity of active chlorine towards nitrogen compounds. No organic intermediates were detected during the electrolytic degradation of cyanuric acid in any the electrolytes, which can be explained by their immediate depletion by hydroxyl radicals produced on the BDD surface. Nitrates and ammonium were the final products of electrolytic degradation of cyanuric acid on BDD anodes in all electrolytes. In addition, small amounts of chloramines were formed in the chloride medium. Low current density (≤10mA/cm(2)) and neutral medium (pH in the range 6-9) should be used for high efficiency electrolytic degradation and negligible formation of hazardous chlorate and perchlorate. PMID:27372125

  12. Arsenic Metabolites, Including N-Acetyl-4-hydroxy-m-arsanilic Acid, in Chicken Litter from a Roxarsone-Feeding Study Involving 1600 Chickens.

    Yang, Zonglin; Peng, Hanyong; Lu, Xiufen; Liu, Qingqing; Huang, Rongfu; Hu, Bin; Kachanoski, Gary; Zuidhof, Martin J; Le, X Chris

    2016-07-01

    The poultry industry has used organoarsenicals, such as 3-nitro-4-hydroxyphenylarsonic acid (Roxarsone, ROX), to prevent disease and to promote growth. Although previous studies have analyzed arsenic species in chicken litter after composting or after application to agricultural lands, it is not clear what arsenic species were excreted by chickens before biotransformation of arsenic species during composting. We describe here the identification and quantitation of arsenic species in chicken litter repeatedly collected on days 14, 24, 28, 30, and 35 of a Roxarsone-feeding study involving 1600 chickens of two strains. High performance liquid chromatography separation with simultaneous detection by both inductively coupled plasma mass spectrometry and electrospray ionization tandem mass spectrometry provided complementary information necessary for the identification and quantitation of arsenic species. A new metabolite, N-acetyl-4-hydroxy-m-arsanilic acid (N-AHAA), was identified, and it accounted for 3-12% of total arsenic. Speciation analyses of litter samples collected from ROX-fed chickens on days 14, 24, 28, 30, and 35 showed the presence of N-AHAA, 3-amino-4-hydroxyphenylarsonic acid (3-AHPAA), inorganic arsenite (As(III)), arsenate (As(V)), monomethylarsonic acid (MMA(V)), dimethylarsinic acid (DMA(V)), and ROX. 3-AHPAA accounted for 3-19% of the total arsenic. Inorganic arsenicals (the sum of As(III) and As(V)) comprised 2-6% (mean 3.5%) of total arsenic. Our results on the detection of inorganic arsenicals, methylarsenicals, 3-AHPAA, and N-AHAA in the chicken litter support recent findings that ROX is actually metabolized by the chicken or its gut microbiome. The presence of the toxic metabolites in chicken litter is environmentally relevant as chicken litter is commonly used as fertilizer. PMID:26876684

  13. Contribution of the lipopolysaccharide to resistance of Shigella flexneri 2a to extreme acidity.

    Mara Martinić

    Full Text Available Shigella flexneri is endemic in most underdeveloped countries, causing diarrheal disease and dysentery among young children. In order to reach its target site, the colon, Shigella must overcome the acid environment of the stomach. Shigella is able to persist in this stressful environment and, because of this ability it can initiate infection following the ingestion of very small inocula. Thus, acid resistance is considered an important virulence trait of this bacterium. It has been reported that moderate acid conditions regulate the expression of numerous components of the bacterial envelope. Because the lipopolysaccharide (LPS is the major component of the bacterial surface, here we have addressed the role of LPS in acid resistance of S. flexneri 2a. Defined deletion mutants in genes encoding proteins involved in the synthesis, assembly and length regulation of the LPS O antigen were constructed and assayed for resistance to pH 2.5 after adaptation to pH 5.5. The results showed that a mutant lacking O antigen was significantly more sensitive to extreme acid conditions than the wild type. Not only the presence of polymerized O antigen, but also a particular polymer length (S-OAg was required for acid resistance. Glucosylation of the O antigen also contributed to this property. In addition, a moderate acidic pH induced changes in the composition of the lipid A domain of LPS. The main modification was the addition of phosphoethanolamine to the 1' phosphate of lipid A. This modification increased resistance of S. flexneri to extreme acid conditions, provide that O antigen was produced. Overall, the results of this work point out to an important role of LPS in resistance of Shigella flexneri to acid stress.

  14. Amino acids, lipid metabolites, and ferritin as potential mediators linking red meat consumption to type 2 diabetes

    Wittenbecher, C.; Muehlenbruch, K.; Kroeger, J.; Jacobs, S; Kuxhaus, O; Floegel, A.; Fritsche, A.; Pischon, T; Prehn, C.; Adamski, J.; Joost, H. G.; Boeing, H.; Schulze, M. B.

    2015-01-01

    BACKGROUND: Habitual red meat consumption was consistently related to a higher risk of type 2 diabetes in observational studies. Potentially underlying mechanisms are unclear. OBJECTIVE: This study aimed to identify blood metabolites that possibly relate red meat consumption to the occurrence of type 2 diabetes. DESIGN: Analyses were conducted in the prospective European Prospective Investigation into Cancer and Nutrition-Potsdam cohort (n = 27,548), applying a nested case-cohort design (n = ...

  15. Population Dynamics and Metabolite Target Analysis of Lactic Acid Bacteria during Laboratory Fermentations of Wheat and Spelt Sourdoughs▿

    Van der Meulen, Roel; Scheirlinck, Ilse; Van Schoor, Ann; Huys, Geert; Vancanneyt, Marc; Vandamme, Peter; De Vuyst, Luc

    2007-01-01

    Four laboratory sourdough fermentations, initiated with wheat or spelt flour and without the addition of a starter culture, were prepared over a period of 10 days with daily back-slopping. Samples taken at all refreshment steps were used for determination of the present microbiota. Furthermore, an extensive metabolite target analysis of more than 100 different compounds was performed through a combination of various chromatographic methods including liquid chromatography-mass spectrometry and...

  16. In vitro inhibition and enhancement of liver microsomal S-777469 metabolism by long-chain fatty acids and serum albumin: insight into in vitro and in vivo discrepancy of metabolite formation in humans.

    Sekiguchi, Kazutaka; Kanazu, Takushi; Murayama, Norie; Yamazaki, Hiroshi; Yamaguchi, Yoshitaka

    2016-06-01

    1. It was previously demonstrated that 10% of S-777469, a cannabinoid receptor 2 selective agonist, is metabolized to its carboxylic acid metabolite (S-777469 5-carboxylic acid, 5-CA) in humans in vivo, while the formation of 5-CA is extremely low in human cryopreserved hepatocytes and liver microsomes (HLMs). In this study, factors causing the different metabolite formation rates of S-777469 in vitro and in vivo were investigated. 2. Formation of 5-CA and S-777469 5-hydroxymethyl (5-HM), a precursor metabolite of 5-CA, was catalyzed by CYP2C9. Arachidonic acid, α-linolenic acid, oleic acid and myristic acid, which have been reported to exist in liver microsomes, inhibited S-777469 oxidation by CYP2C9, but serum albumin enhanced this reactions. 3. The IC50 values of these fatty acids for 5-CA formation from 5-HM were lower than those of 5-HM formation from S-777469. Serum albumin extensively enhanced 5-CA formation from 5-HM in comparison to 5-HM formation from S-777469. 4. CYP2C9 was the enzyme responsible for S-777469 oxidation in human livers. The suppressive effects of several fatty acids and enhancing action of serum albumin in vitro are likely to be the causal factors for the apparently different rates of in vitro and in vivo metabolite formation of S-777469. PMID:26677906

  17. A novel study of screening and confirmation of modafinil, adrafinil and their metabolite modafinilic acid under EI-GC-MS and ESI-LC-MS-MS ionization

    Dubey S

    2009-01-01

    Full Text Available Objective: Adrafinil and modafinil have received wide publicity and have become controversial in the sporting world when several athletes were discovered allegedly using these drugs as doping agents. By acknowledging the facts, the World Anti-Doping Agency (WADA banned these drugs in sports since 2004. The present study explores the possibility of differentiating adrafinil and modafinil and their major metabolites under electron impact ionization in gas chromatograph-mass spectrometer (GC-MSD and electrospray ionization in liquid chromatograph-mass spectrometer (LC-MS/MS by studying the fragmentation pattern of these drugs. Materials and Methods: Adrafinil, modafinil and their major metabolite, modafinilic acid were analyzed on EI-GC-MSD and ESI-LC-MS/MS using various individual parameters on both the instruments. The analytical technique and equipment used in the analysis were an Agilent 6890N GC with 5973 mass selective detector for the GC-MSD analysis and an Agilent 1100 HPLC with API-3200 Triple quadrupole mass spectrometer for the LC-MS/MS analysis. Validation of both methods was performed using six replicates at different concentrations. Result and Discussion: The results show that adrafinil, modafinil and their major metabolite modafinilic acid could be detected as a single artifact without differentiation under EI-GC-MSD analysis. However, all drugs could be detected and differentiated under ESI-LCMS/MS analysis without any artifaction. The GC-MSD analysis gives a single artifact for both the drugs without differentiation and thus can be used as a marker for screening purposes. Further, the Multiple Reaction Monitoring (MRM method developed under LC-MS/MS is fit for the purpose for confirmation of suspicious samples in routine sports testing and in forensic and clinical analysis.

  18. Identification of metabolites with anticancer properties by computational metabolomics

    Bowen Nathan J

    2008-06-01

    Full Text Available Abstract Background Certain endogenous metabolites can influence the rate of cancer cell growth. For example, diacylglycerol, ceramides and sphingosine, NAD+ and arginine exert this effect by acting as signaling molecules, while carrying out other important cellular functions. Metabolites can also be involved in the control of cell proliferation by directly regulating gene expression in ways that are signaling pathway-independent, e.g. by direct activation of transcription factors or by inducing epigenetic processes. The fact that metabolites can affect the cancer process on so many levels suggests that the change in concentration of some metabolites that occurs in cancer cells could have an active role in the progress of the disease. Results CoMet, a fully automated Computational Metabolomics method to predict changes in metabolite levels in cancer cells compared to normal references has been developed and applied to Jurkat T leukemia cells with the goal of testing the following hypothesis: Up or down regulation in cancer cells of the expression of genes encoding for metabolic enzymes leads to changes in intracellular metabolite concentrations that contribute to disease progression. All nine metabolites predicted to be lowered in Jurkat cells with respect to lymphoblasts that were examined (riboflavin, tryptamine, 3-sulfino-L-alanine, menaquinone, dehydroepiandrosterone, α-hydroxystearic acid, hydroxyacetone, seleno-L-methionine and 5,6-dimethylbenzimidazole, exhibited antiproliferative activity that has not been reported before, while only two (bilirubin and androsterone of the eleven tested metabolites predicted to be increased or unchanged in Jurkat cells displayed significant antiproliferative activity. Conclusion These results: a demonstrate that CoMet is a valuable method to identify potential compounds for experimental validation, b indicate that cancer cell metabolism may be regulated to reduce the intracellular concentration of

  19. Characterization and analysis of the cotton cyclopropane fatty acid synthase family and their contribution to cyclopropane fatty acid synthesis

    Yu X. H.; Shanklin J.; Rawat, R.

    2011-05-01

    Cyclopropane fatty acids (CPA) have been found in certain gymnosperms, Malvales, Litchi and other Sapindales. The presence of their unique strained ring structures confers physical and chemical properties characteristic of unsaturated fatty acids with the oxidative stability displayed by saturated fatty acids making them of considerable industrial interest. While cyclopropenoid fatty acids (CPE) are well-known inhibitors of fatty acid desaturation in animals, CPE can also inhibit the stearoyl-CoA desaturase and interfere with the maturation and reproduction of some insect species suggesting that in addition to their traditional role as storage lipids, CPE can contribute to the protection of plants from herbivory. Three genes encoding cyclopropane synthase homologues GhCPS1, GhCPS2 and GhCPS3 were identified in cotton. Determination of gene transcript abundance revealed differences among the expression of GhCPS1, 2 and 3 showing high, intermediate and low levels, respectively, of transcripts in roots and stems; whereas GhCPS1 and 2 are both expressed at low levels in seeds. Analyses of fatty acid composition in different tissues indicate that the expression patterns of GhCPS1 and 2 correlate with cyclic fatty acid (CFA) distribution. Deletion of the N-terminal oxidase domain lowered GhCPS's ability to produce cyclopropane fatty acid by approximately 70%. GhCPS1 and 2, but not 3 resulted in the production of cyclopropane fatty acids upon heterologous expression in yeast, tobacco BY2 cell and Arabidopsis seed. In cotton GhCPS1 and 2 gene expression correlates with the total CFA content in roots, stems and seeds. That GhCPS1 and 2 are expressed at a similar level in seed suggests both of them can be considered potential targets for gene silencing to reduce undesirable seed CPE accumulation. Because GhCPS1 is more active in yeast than the published Sterculia CPS and shows similar activity when expressed in model plant systems, it represents a strong candidate gene

  20. Simultaneous Determination and Pharmacokinetic Study of Protocatechuic Aldehyde and Its Major Active Metabolite Protocatechuic Acid in Rat Plasma by Liquid Chromatography-Tandem Mass Spectrometry.

    Wang, Xiangyang; Yan, Kaijing; Ma, Xiaohui; Li, Wei; Chu, Yang; Guo, Jiahua; Li, Shuming; Zhou, Shuiping; Zhu, Yonghong; Liu, Changxiao

    2016-05-01

    A very simple and selective high-performance liquid chromatography electrospray ionization tandem mass spectrometry (LC-MS-MS) method was developed for simultaneous determination and pharmacokinetic study of protocatechuic aldehyde (PAL) and its active metabolite protocatechuic acid (PCA). The method involves a simple liquid-liquid extraction with ethyl acetate. The separation was performed on a Hypersil GOLD C18column (2.1 × 150 mm, 3.0 µm; particle, Thermo, USA) with isocratic elution using a mobile phase consisted of methanol and water (containing 0.1% formic acid) at a flow rate of 0.2 mL/min. The detection of target compounds was done by using low-energy collision dissociation tandem mass spectrometry (CID-MS-MS) using the selective reaction monitoring scan mode. The method was linear for all analytes over the investigated range for all correlation coefficients greater than 0.9950. The lower limits of quantification were 2.0 ng/mL for PAL and PCA. The intra- and interday precisions (relative standard deviation, RSD %) were <6.84 and 5.54%, and the accuracy (relative error, RE %) was between -2.85 and 0.74% (n= 6). The developed method was applied to study the pharmacokinetics of PAL and its major active metabolite PCA in rat plasma after oral and intravenous administration of PAL. PMID:26969682

  1. Liquid chromatography-electrospray ionization-tandem mass spectrometry for simultaneous analysis of chlorogenic acids and their metabolites in human plasma.

    Matsui, Yuji; Nakamura, Shun; Kondou, Naoki; Takasu, Yoshio; Ochiai, Ryuji; Masukawa, Yoshinori

    2007-10-15

    A method using liquid chromatography-electrospray ionization-tandem mass spectrometry (LC-ESI-MS/MS) was developed for the simultaneous analysis of nine chlorogenic acids (CGAs), three isomers each of caffeoylquinic acids (CQAs), feruloylquinic acids (FQAs) and dicaffeoylquinic acids (dCQAs), and their two metabolites, caffeic acid (CA) and ferulic acid (FA), in human plasma. In simultaneous multiple reaction monitoring (MRM) measurements using ESI-MS/MS with a negative ion mode, a deprotonated molecular ion derived from each of the 11 molecules was used as a precursor ion while three diagnostic product ions characteristic for each were selected for the qualitative analysis. To obtain maximal intensities for all diagnostic product ions, the collision energy was optimized for each one. LC separation was achieved under conditions of a reversed-phase Inertsil ODS-2 column combined with a gradient elution system using 50mM acetic acid with 3% acetonitrile aqueous solution and 50 mM acetic acid with 100% acetonitrile. In the quantitative analysis, one of the three diagnostic product ions for each of the 11 molecules was selected. Application of simultaneous LC-ESI-MS/MS MRM measurements to analyze the 11 standards spiked into blank human plasma indicated that all diagnostic product ions were detected without any interference, and that the sensitivity, linearity and recovery of this method were acceptable. When using this method to analyze those 11 molecules in the plasma after oral ingestion of 250 ml of a drink containing a green coffee bean extract (300 mg CGAs), all 11 molecules were identified and CQAs, FQAs and FA were quantified. CQAs, FQAs and dCQAs in human plasma were detected for the first time. This method should be useful to understand the biological and pharmacological effects of CGAs, such as improvement of human hypertension. PMID:17766198

  2. Induction of CYP26A1 by Metabolites of Retinoic Acid: Evidence That CYP26A1 Is an Important Enzyme in the Elimination of Active Retinoids

    Topletz, Ariel R.; Tripathy, Sasmita; Foti, Robert S.; Shimshoni, Jakob A.; Nelson, Wendel L.

    2015-01-01

    All-trans-retinoic acid (atRA), the active metabolite of vitamin A, induces gene transcription via binding to nuclear retinoic acid receptors (RARs). The primary hydroxylated metabolites formed from atRA by CYP26A1, and the subsequent metabolite 4-oxo-atRA, bind to RARs and potentially have biologic activity. Hence, CYP26A1, the main atRA hydroxylase, may function either to deplete bioactive retinoids or to form active metabolites. This study aimed to determine the role of CYP26A1 in modulating RAR activation via formation and elimination of active retinoids. After treatment of HepG2 cells with atRA, (4S)-OH-atRA, (4R)-OH-atRA, 4-oxo-atRA, and 18-OH-atRA, mRNAs of CYP26A1 and RARβ were increased 300- to 3000-fold, with 4-oxo-atRA and atRA being the most potent inducers. However, >60% of the 4-OH-atRA enantiomers were converted to 4-oxo-atRA in the first 12 hours of treatment, suggesting that the activity of the 4-OH-atRA was due to 4-oxo-atRA. In human hepatocytes, atRA, 4-OH-atRA, and 4-oxo-atRA induced CYP26A1 and 4-oxo-atRA formation was observed from 4-OH-atRA. In HepG2 cells, 4-oxo-atRA formation was observed even in the absence of CYP26A1 activity and this formation was not inhibited by ketoconazole. In human liver microsomes, 4-oxo-atRA formation was supported by NAD+, suggesting that 4-oxo-atRA formation is mediated by a microsomal alcohol dehydrogenase. Although 4-oxo-atRA was not formed by CYP26A1, it was depleted by CYP26A1 (Km = 63 nM and intrinsic clearance = 90 μl/min per pmol). Similarly, CYP26A1 depleted 18-OH-atRA and the 4-OH-atRA enantiomers. These data support the role of CYP26A1 to clear bioactive retinoids, and suggest that the enzyme forming active 4-oxo-atRA may be important in modulating retinoid action. PMID:25492813

  3. Metabolites from roots of Colubrina greggii var. yucatanensis and evaluation of their antiprotozoan, cytotoxic and antiproliferative activities

    Purification of the root extract of Colubrina greggii var. yucatanensis resulted in the isolation and identification of 3-O-acetyl ceanothic acid as a new natural ceanothane triterpene, together with the known metabolites ceanothic acid, cenothenic acid, betulinic acid, discarine B and chrysophanein. The natural products and the semisynthetic esters acetyl dimethyl ceanothate, dimethyl ceanothate and chrysophanein peracetate showed moderate to low leishmanicidal and trypanocidal activities. None of the metabolites showed cytotoxic or antiproliferative effects. The results also suggested that betulinic acid contributes to the antiplasmodial activity originally detected in the crude root extract of C. greggii var. yucatanensis. (author)

  4. Metabolites from roots of Colubrina greggii var. yucatanensis and evaluation of their antiprotozoan, cytotoxic and antiproliferative activities

    Dominguez-Carmona, Dafne B.; Escalante-Erosa, Fabiola; Garcia-Sosa, Karlina; Pena-Rodriguez, Luis M., E-mail: lmanuel@cicy.m [Centro de Investigacion Cientifica de Yucatan (Mexico). Unidad de Biotecnologia; Ruiz-Pinell, Grace; Gutierrez-Yapu, David; Gimenez-Turba, Alberto [Universidad Mayor de San Andres, La Paz (Bolivia, Plurinational State of). Inst. de Investigaciones Farmaco-Bioquimicas; Chan-Bacab, Manuel J. [Universidad Autonoma de Campeche (Mexico). Dept. de Microbiologia Ambiental y Biotecnologia; Moo-Puc, Rosa E. [Centro Medico Ignacio Garcia Tellez, Col. Industrial, Merida, Yucatan (Mexico). Unidad de Investigacion Medica Yucatan y Unidad Medica de Alta Especialidad; Veitch, Nigel C. [Jodrell Laboratory, Richmond, Surrey (United Kingdom)

    2011-07-01

    Purification of the root extract of Colubrina greggii var. yucatanensis resulted in the isolation and identification of 3-O-acetyl ceanothic acid as a new natural ceanothane triterpene, together with the known metabolites ceanothic acid, cenothenic acid, betulinic acid, discarine B and chrysophanein. The natural products and the semisynthetic esters acetyl dimethyl ceanothate, dimethyl ceanothate and chrysophanein peracetate showed moderate to low leishmanicidal and trypanocidal activities. None of the metabolites showed cytotoxic or antiproliferative effects. The results also suggested that betulinic acid contributes to the antiplasmodial activity originally detected in the crude root extract of C. greggii var. yucatanensis. (author)

  5. Protecting Cell Walls from Binding Aluminum by Organic Acids Contributes to Aluminum Resistance

    Ya-Ying Li; Yue-Jiao Zhang; Yuan Zhou; Jian-Li Yang; Shao-Jian Zheng

    2009-01-01

    Aluminum-induced secretion of organic acids from the root apex has been demonstrated to be one major AI resistance mechanism in plants. However, whether the organic acid concentration is high enough to detoxify AI in the growth medium is frequently questioned. The genotypes of Al-resistant wheat, Cassia tora L. and buckwheat secrete malate, citrate and oxalate, respectively. In the present study we found that at a 35% inhibition of root elongation, the AI activities in the solution were 10, 20, and 50 μM with the corresponding malate, citrate, and oxalate exudation at the rates of 15, 20 and 21 nmol/cm2 per 12 h, respectively, for the above three plant species. When exogenous organic acids were added to ameliorate Al toxicity, twofold and eightfold higher oxalate and malate concentrations were required to produce the equal effect by citrate. After the root apical cell walls were isolated and preincubated in 1 mM malate, oxalate or citrate solution overnight, the total amount of AI adsorbed to the cell walls all decreased significantly to a similar level, implying that these organic acids own an equal ability to protect the cell walls from binding AI. These findings suggest that protection of cell walls from binding Al by organic acids may contribute significantly to AI resistance.

  6. Evaluation of the stereoselective biotransformation of permethrin in human liver microsomes: contributions of cytochrome P450 monooxygenases to the formation of estrogenic metabolites.

    Lavado, Ramon; Li, Jiwen; Rimoldi, John M; Schlenk, Daniel

    2014-04-21

    Permethrin (PM) is a pyrethroid insecticide that exists as 4 enantiomers. Biotransformation of PM to estrogen receptor agonists (3-phenoxybenzyl alcohol (PBOH) and 3-(4'-hydroxyphenoxy)-benzyl alcohol (3,4 PBOH)) has been shown to be stereoselective in other vertebrate species. This study evaluated the biotransformation of PM enantiomers in human liver microsomes and with recombinant CYP3A4 and CYP2C19. PBOH and 3,4 PBOH were the only metabolites detected from in vitro incubations including each of the 4 enantiomers of PM with 1R-trans PM having the most efficient NADPH-catalyzed biotransformation to both metabolites. Coincubation with the CYP inhibitor ketoconazole and time course experiments with liver microsomes and recombinant CYP2C19 and CYP3A4 indicated CYP-catalyzed stereoselective cleavage of the ester followed by 4-hydoxylation to 3,4' PBOH. These data indicate potential dispositional differences may occur with PM enantiomers and a shift in putative molecular targets. While cleavage of pyrethroid esters lead to detoxification of the acute neurological effects, formation of the benzyl alcohol and hydroxylated metabolite may lead to estrogenic responses, since each of these metabolites are estrogen receptor ligands. PMID:24548679

  7. Contribution of glutamate decarboxylase in Lactobacillus reuteri to acid resistance and persistence in sourdough fermentation

    Gänzle Michael G; Schlicht Sabine; Su Marcia S

    2011-01-01

    Abstract Background Acid stress impacts the persistence of lactobacilli in industrial sourdough fermentations, and in intestinal ecosystems. However, the contribution of glutamate to acid resistance in lactobacilli has not been demonstrated experimentally, and evidence for the contribution of acid resistance to the competitiveness of lactobacilli in sourdough is lacking. It was therefore the aim of this study to investigate the ecological role of glutamate decarboxylase in L. reuteri. Results...

  8. The enteric bacterial metabolite propionic acid alters brain and plasma phospholipid molecular species: further development of a rodent model of autism spectrum disorders

    Thomas Raymond H

    2012-07-01

    Full Text Available Abstract Gastrointestinal symptoms and altered blood phospholipid profiles have been reported in patients with autism spectrum disorders (ASD. Most of the phospholipid analyses have been conducted on the fatty acid composition of isolated phospholipid classes following hydrolysis. A paucity of information exists on how the intact phospholipid molecular species are altered in ASD. We applied ESI/MS to determine how brain and blood intact phospholipid species were altered during the induction of ASD-like behaviors in rats following intraventricular infusions with the enteric bacterial metabolite propionic acid. Animals were infused daily for 8 days, locomotor activity assessed, and animals killed during the induced behaviors. Propionic acid infusions increased locomotor activity. Lipid analysis revealed treatment altered 21 brain and 30 blood phospholipid molecular species. Notable alterations were observed in the composition of brain SM, diacyl mono and polyunsaturated PC, PI, PS, PE, and plasmalogen PC and PE molecular species. These alterations suggest that the propionic acid rat model is a useful tool to study aberrations in lipid metabolism known to affect membrane fluidity, peroxisomal function, gap junction coupling capacity, signaling, and neuroinflammation, all of which may be associated with the pathogenesis of ASD.

  9. Pomegranate Juice Metabolites, Ellagic Acid and Urolithin A, Synergistically Inhibit Androgen-Independent Prostate Cancer Cell Growth via Distinct Effects on Cell Cycle Control and Apoptosis

    Roberto Vicinanza

    2013-01-01

    Full Text Available Ellagitannins (ETs from pomegranate juice (PJ are bioactive polyphenols with chemopreventive potential against prostate cancer (PCa. ETs are not absorbed intact but are partially hydrolyzed in the gut to ellagic acid (EA. Colonic microflora can convert EA to urolithin A (UA, and EA and UA enter the circulation after PJ consumption. Here, we studied the effects of EA and UA on cell proliferation, cell cycle, and apoptosis in DU-145 and PC-3 androgen-independent PCa cells and whether combinations of EA and UA affected cell proliferation. EA demonstrated greater dose-dependent antiproliferative effects in both cell lines compared to UA. EA induced cell cycle arrest in S phase associated with decreased cyclin B1 and cyclin D1 levels. UA induced a G2/M arrest and increased cyclin B1 and cdc2 phosphorylation at tyrosine-15, suggesting inactivation of the cyclin B1/cdc2 kinase complex. EA induced apoptosis in both cell lines, while UA had a less pronounced proapoptotic effect only in DU-145. Cotreatment with low concentrations of EA and UA dramatically decreased cell proliferation, exhibiting synergism in PC-3 cells evaluated by isobolographic analysis and combination index. These data provide information on pomegranate metabolites for the prevention of PCa recurrence, supporting the role of gut flora-derived metabolites for cancer prevention.

  10. Salicylic acid treatment reduces the rot of postharvest citrus fruit by inducing the accumulation of H2O2, primary metabolites and lipophilic polymethoxylated flavones.

    Zhu, Feng; Chen, Jiajing; Xiao, Xue; Zhang, Mingfei; Yun, Ze; Zeng, Yunliu; Xu, Juan; Cheng, Yunjiang; Deng, Xiuxin

    2016-09-15

    To comprehensively analyze the effects of salicylic acid (SA) on the storability of Satsuma mandarin (Citrus unshiu), fruits were treated with 2mM SA. The disease incidence of control/SA-treated fruit at 50d and 120d after treatment was 23.3%/10% and 67.3%/23.3%, respectively, suggesting that SA treatment can significantly reduce the rot rate of postharvest citrus fruit. Fruit quality assays revealed that the treatment can maintain fruit firmness without affecting the inner quality. Furthermore, the contents of H2O2 and some defense-related metabolites, such as ornithine and threonine, in citrus pericarp, were significantly increased by SA treatment. Moreover, it was lipophilic polymethoxylated flavones, rather than flavanone glycosides, that accumulated in SA-treated fruits and these can directly inhibit pathogen development. These results suggest that the effects of SA on postharvest citrus fruit may be attributed to the accumulation of H2O2 and defense-related metabolites. PMID:27080881

  11. Asymmetric functional contributions of acidic and aromatic side chains in sodium channel voltage-sensor domains

    Pless, Stephan Alexander; Elstone, Fisal D; Niciforovic, Ana P; Galpin, Jason D; Yang, Runying; Kurata, Harley T; Ahern, Christopher A

    2014-01-01

    largely enigmatic. To this end, natural and unnatural side chain substitutions were made in the S2 hydrophobic core (HC), the extracellular negative charge cluster (ENC), and the intracellular negative charge cluster (INC) of the four VSDs of the skeletal muscle sodium channel isoform (NaV1.4). The......Voltage-gated sodium (NaV) channels mediate electrical excitability in animals. Despite strong sequence conservation among the voltage-sensor domains (VSDs) of closely related voltage-gated potassium (KV) and NaV channels, the functional contributions of individual side chains in Nav VSDs remain...... functional phenotypes that are different from those observed previously in Kv VSDs. In contrast, and similar to results obtained with Kv channels, individually neutralizing acidic side chains with synthetic derivatives and with natural amino acid substitutions in the INC had little or no effect on the...

  12. Contribution of Indole-3-Acetic Acid Production to the Epiphytic Fitness of Erwinia herbicola

    Brandl, M. T.; Lindow, S E

    1998-01-01

    Erwinia herbicola 299R produces large quantities of indole-3-acetic acid (IAA) in culture media supplemented with l-tryptophan. To assess the contribution of IAA production to epiphytic fitness, the population dynamics of the wild-type strain and an IAA-deficient mutant of this strain on leaves were studied. Strain 299XYLE, an isogenic IAA-deficient mutant of strain 299R, was constructed by insertional interruption of the indolepyruvate decarboxylase gene of strain 299R with the xylE gene, wh...

  13. High throughput HPLC-ESI(-)-MS/MS methodology for mercapturic acid metabolites of 1,3-butadiene: Biomarkers of exposure and bioactivation.

    Kotapati, Srikanth; Esades, Amanda; Matter, Brock; Le, Chap; Tretyakova, Natalia

    2015-11-01

    1,3-Butadiene (BD) is an important industrial and environmental carcinogen present in cigarette smoke, automobile exhaust, and urban air. The major urinary metabolites of BD in humans are 2-(N-acetyl-L-cystein-S-yl)-1-hydroxybut-3-ene/1-(N-acetyl-L-cystein-S-yl)-2-hydroxybut-3-ene (MHBMA), 4-(N-acetyl-L-cystein-S-yl)-1,2-dihydroxybutane (DHBMA), and 4-(N-acetyl-L-cystein-S-yl)-1,2,3-trihydroxybutyl mercapturic acid (THBMA), which are formed from the electrophilic metabolites of BD, 3,4-epoxy-1-butene (EB), hydroxymethyl vinyl ketone (HMVK), and 3,4-epoxy-1,2-diol (EBD), respectively. In the present work, a sensitive high-throughput HPLC-ESI(-)-MS/MS method was developed for simultaneous quantification of MHBMA and DHBMA in small volumes of human urine (200 μl). The method employs a 96 well Oasis HLB SPE enrichment step, followed by isotope dilution HPLC-ESI(-)-MS/MS analysis on a triple quadrupole mass spectrometer. The validated method was used to quantify MHBMA and DHBMA in urine of workers from a BD monomer and styrene-butadiene rubber production facility (40 controls and 32 occupationally exposed to BD). Urinary THBMA concentrations were also determined in the same samples. The concentrations of all three BD-mercapturic acids and the metabolic ratio (MHBMA/(MHBMA+DHBMA+THBMA)) were significantly higher in the occupationally exposed group as compared to controls and correlated with BD exposure, with each other, and with BD-hemoglobin biomarkers. This improved high throughput methodology for MHBMA and DHBMA will be useful for future epidemiological studies in smokers and occupationally exposed workers. PMID:25727266

  14. nC60 deposition kinetics: the complex contribution of humic acid, ion concentration, and valence.

    McNew, Coy P; LeBoeuf, Eugene J

    2016-07-01

    The demonstrated toxicity coupled with inevitable environmental release of nC60 raise serious concerns about its environmental fate and transport, therefore it is crucial to understand how nC60 will interact with subsurface materials including attached phase soil and sediment organic matter (AP-SOM). This study investigated the attachment of nC60 onto a Harpeth humic acid (HHA) coated silica surface under various solution conditions using a quartz crystal microbalance with dissipation monitoring. The HHA coating greatly enhanced nC60 attachment at low ion concentrations while hindering attachment at high ion concentrations in the presence of both mono and divalent cations. At low ion concentrations, the HHA greatly reduced the surface potential of the silica, enhancing nC60 deposition through reduction in the electrostatic repulsion. At high ion concentrations however, the reduced surface potential became less important due to the near zero energy barrier to deposition and therefore non-DLVO forces dominated, induced by compaction of the HHA layer, and leading to hindered attachment. In this manner, observed contributions from the HHA layer were more complex than previously reported and by monitoring surface charge and calculated DLVO interaction energy alongside attachment experiments, this study advances the mechanistic understanding of the variable attachment contributions from the humic acid layer. PMID:27061365

  15. Recent advances in biosynthesis of fatty acids derived products in Saccharomyces cerevisiae via enhanced supply of precursor metabolites.

    Lian, Jiazhang; Zhao, Huimin

    2015-03-01

    Fatty acids or their activated forms, fatty acyl-CoAs and fatty acyl-ACPs, are important precursors to synthesize a wide variety of fuels and chemicals, including but not limited to free fatty acids (FFAs), fatty alcohols (FALs), fatty acid ethyl esters (FAEEs), and alkanes. However, Saccharomyces cerevisiae, an important cell factory, does not naturally accumulate fatty acids in large quantities. Therefore, metabolic engineering strategies were carried out to increase the glycolytic fluxes to fatty acid biosynthesis in yeast, specifically to enhance the supply of precursors, eliminate competing pathways, and bypass the host regulatory network. This review will focus on the genetic manipulation of both structural and regulatory genes in each step for fatty acids overproduction in S. cerevisiae, including from sugar to acetyl-CoA, from acetyl-CoA to malonyl-CoA, and from malonyl-CoA to fatty acyl-CoAs. The downstream pathways for the conversion of fatty acyl-CoAs to the desired products will also be discussed. PMID:25306882

  16. Cellular responses induced in vitro by pestheic acid, a fungal metabolite, in a gastric adenocarcinoma cell line (PG100).

    Sousa, J M C; Matos, L A; Alcântara, D F A; Ribeiro, H F; Santos, L S; Oliveira, M N; Brito-Junior, L C; Khayat, A S; Guimarães, A C; Cunha, L A; Burbano, R R; Bahia, M O

    2013-01-01

    There is a constant search for new cancer treatments that are less aggressive and economically affordable. In this context, natural products extracted from plants, fungi, and microorganisms are of great interest. Pestheic acid, or dihidromaldoxin, is a chlorinated diphenylic ether extracted from the phytopathogenic fungus Pestalotiopsis guepinii (Amphisphaeriaceae). We assessed the cytotoxic, cytostatic, and genotoxic effects of pestheic acid in a gastric adenocarcinoma cell line (PG100). A decrease in clonogenic survival was observed. Pestheic acid also induced significant increases in both micronucleus and nucleoplasmic bridge frequency. However, we did not observe changes in cell cycle kinetics or apoptosis induction. Reactive oxygen species induced by diphenylic ethers may explain the genotoxicity and mutagenicity of pestheic acid. The absence of repair checkpoints that we observed is probably due to the fact that the PG100 cell line lacks the TP53 gene, which is common in gastric cancers. Even though pestheic acid has had a clear cytotoxic effect, the minimal inhibitory concentration was high, which shows that pestheic acid is not an active anticancer compound under these conditions. PMID:24114206

  17. Voluntary fortification of breakfast cereals with folic acid: contribution to dietary intake in Australia.

    Japar, Devina; Chandra-Hioe, Maria V; Shrestha, Ashok; Arcot, Jayashree

    2016-05-01

    Ready-to-eat breakfast cereals have been voluntarily fortified with folic acid since 1995, with the purpose of reducing the prevalence of neural tube defects in utero. Using data from the recent Australian Health Survey, this study aimed to estimate folate intake from one serving of breakfast cereals (median amount). Various commercial brands were purchased in 2002 (n = 19) and in 2014 (n = 14); folate was determined by microbiological assay and high-pressure liquid chromatography (HPLC). Total folate (μg/100 g) in 2002 and 2014 selections were 144-633 and 147-564, respectively, and mostly comparable to nutrition labels. Folic acid (2014 selection) using HPLC, ranged from 85 to 411 μg/100 g. Intake of 51 g cereals/serving by individuals ≥2 years could contribute 75-288 μg dietary folate equivalent. It seems that folic acid intake among children (2-3 years) exceeds the recommended dietary intake, when certain brands of breakfast cereals are consumed. Accordingly, the benefits and potential detrimental effects of the voluntary fortification need to be further explored. PMID:26903206

  18. Effect of heat stress on the serum concentrations of free amino acids and some of their metabolites in growing pigs.

    Morales, A; Cota, S E M; Ibarra, N O; Arce, N; Htoo, J K; Cervantes, M

    2016-07-01

    Exposure to heat stress (HS) may affect the intestinal epithelia of pigs, resulting in impaired digestive and absorptive capacity. The serum concentration (SC) of free AA in pigs can be used as indicators of their availability. This study was conducted with 12 crossbred (Landrace × Hampshire × Duroc) pigs (29.0 ± 2.8 kg initial BW) distributed into 2 groups to analyze the SC of free AA and some AA metabolites in pigs exposed to HS conditions. The treatments were pigs housed under natural HS conditions in a room with no ambient temperature control (23.6 to 37.6°C, HS) and pigs housed at thermoneutral conditions (24 ± 2°C), feed restricted to a level similar to that of their HS counterparts. All pigs received a wheat-soybean meal diet. Blood samples were collected at both the absorptive (2.5 h after a meal) and postabsorptive (10.0 h after a meal) phase. At the absorptive phase, the SC of free Arg, Leu, Lys, Phe, Thr, and Trp were lower ( carnosine, ornithine (Orn), and Tau were lower ( carnosine and sarcosine ( < 0.05) decreased in HS pigs. The results of this study show a marked and differential effect of HS on the SC of AA. These data indicate that HS negatively affects the digestive and absorptive capacity of pigs and that the metabolism of some AA is modified in pigs to counteract the negative effects of the HS. PMID:27482670

  19. Whey Protein Delays Gastric Emptying and Suppresses Plasma Fatty Acids and Their Metabolites Compared to Casein, Gluten, and Fish Protein

    Stanstrup, Jan; Schou, Simon S; Holmer-Jensen, Jens;

    2014-01-01

    studies, the WI meal caused a decreased rate of gastric emptying compared to the other test meals. The WI meal also caused elevated levels of a number of amino acids, possibly stimulating insulin release leading to reduced plasma glucose. The WI meal also caused decreased levels of a number of fatty acids......Whey protein has been demonstrated to improve fasting lipid and insulin response in overweight and obese individuals. To establish new hypotheses for this effect and to investigate the impact of stomach emptying, we compared plasma profiles after intake of whey isolate (WI), casein, gluten (GLU...

  20. LC-MS and GC-MS metabolite profiling of nickel(II) complexes in the latex of the nickel-hyperaccumulating tree Sebertia acuminata and identification of methylated aldaric acid as a new nickel(II) ligand

    Callahan, D. L.; Roessner, U.; Dumontet, V.; Perrier, Nicolas; Wedd, A. G.; O'HAIR, R A J; Baker, A. J. M.; Kolev, S. D.

    2008-01-01

    Targeted liquid chromatography mass spectrometry (LC-MS) technology using size exclusion chromatography and metabolite profiling based on gas chromatography-mass spectrometry (GC-MS) were used to study the nickel-rich latex of the hyperaccumulating tree Sebertia acuminata. More than 120 compounds were detected, 57 of these were subsequently identified. A methylated aldaric acid (2,4,5-trihydroxy-3-methoxy-1,6-hexan-dioic acid) was identified for the first time in biological extracts and its s...

  1. The contribution of SNAT1 to system A amino acid transporter activity in human placental trophoblast

    Research highlights: → mRNA levels for SNAT1 are higher than other system A subtype mRNAs in primary human cytotrophoblast. → SNAT1 knockdown in cytotrophoblast cells significantly reduces system A activity. → SNAT1 is a key contributor to system A-mediated amino acid transport in human placenta. -- Abstract: System A-mediated amino acid transport across the placenta is important for the supply of neutral amino acids needed for fetal growth. All three system A subtypes (SNAT1, 2, and 4) are expressed in human placental trophoblast suggesting there is an important biological role for each. Placental system A activity increases as pregnancy progresses, coinciding with increased fetal nutrient demands. We have previously shown SNAT4-mediated system A activity is higher in first trimester than at term, suggesting that SNAT1 and/or SNAT2 are responsible for the increased system A activity later in gestation. However, the relative contribution of each subtype to transporter activity in trophoblast at term has yet to be evaluated. The purpose of this study was to identify the predominant subtype of system A in cytotrophoblast cells isolated from term placenta, maintained in culture for 66 h, by: (1) measuring mRNA expression of the three subtypes and determining the Michaelis-Menten constants for uptake of the system A-specific substrate, 14C-MeAIB, (2) investigating the contribution of SNAT1 to total system A activity using siRNA. Results: mRNA expression was highest for the SNAT1 subtype of system A. Kinetic analysis of 14C-MeAIB uptake revealed two distinct transport systems; system 1: Km = 0.38 ± 0.12 mM, Vmax = 27.8 ± 9.0 pmol/mg protein/20 min, which resembles that reported for SNAT1 and SNAT2 in other cell types, and system 2: Km = 45.4 ± 25.0 mM, Vmax = 1190 ± 291 pmol/mg protein/20 min, which potentially represents SNAT4. Successful knockdown of SNAT1 mRNA using target-specific siRNA significantly reduced system A activity (median 75% knockdown, n = 7

  2. Physiological responses of glyphosate-resistant and glyphosate-sensitive soybean to aminomethylphosphonic acid a metabolite of glyphosate

    Aminomethylphosphonic acid (AMPA) is formed in glyphosate-treated glyphosate-resistant (GR) and glyphosate-sensitive (GS) soybean [Glycine max (L.) Merr.] plants and is known to cause yellowing in soybean. Although, AMPA is less phytotoxic than glyphosate, its mode of action is different from that o...

  3. Beyond prostaglandins - chemistry and biology of cyclic oxygenated metabolites formed by free-radical pathways from polyunsaturated fatty acids

    Jahn, Ullrich; Galano, J. M.; Durand, T.

    2008-01-01

    Roč. 47, č. 32 (2008), s. 5894-5955. ISSN 1433-7851 Institutional research plan: CEZ:AV0Z40550506 Keywords : biological activity * fatty acids * isoprostanes * oxidation * total synthesis Subject RIV: CC - Organic Chemistry Impact factor: 10.879, year: 2008

  4. Abscisic Acid Induced Changes in Production of Primary and Secondary Metabolites, Photosynthetic Capacity, Antioxidant Capability, Antioxidant Enzymes and Lipoxygenase Inhibitory Activity of Orthosiphon stamineus Benth.

    Mohd Hafiz Ibrahim

    2013-07-01

    Full Text Available An experiment was conducted to investigate and distinguish the relationships in the production of total phenolics, total flavonoids, soluble sugars, H2O2, O2−, phenylalanine ammonia lyase (PAL activity, leaf gas exchange, antioxidant activity, antioxidant enzyme activity [ascorbate peroxidase (APX, catalase (CAT, superoxide dismutase (SOD and Lipoxygenase inhibitory activity (LOX] under four levels of foliar abscisic acid (ABA application (0, 2, 4, 6 µM for 15 weeks in Orthosiphon stamineus Benth. It was found that the production of plant secondary metabolites, soluble sugars, antioxidant activity, PAL activity and LOX inhibitory activity was influenced by foliar application of ABA. As the concentration of ABA was increased from 0 to 6 µM the production of total phenolics, flavonoids, sucrose, H2O2, O2−, PAL activity and LOX inhibitory activity was enhanced. It was also observed that the antioxidant capabilities (DPPH and ORAC were increased. This was followed by increases in production of antioxidant enzymes APX, CAT and SOD. Under high application rates of ABA the net photosynthesis and stomatal conductance was found to be reduced. The production of primary and secondary metabolites displayed a significant positive relationship with H2O2 (total phenolics, r2 = 0.877; total flavonoids, r2 = 0.812; p ≤ 0.05 and O2− (total phenolics, r2 = 0.778; total flavonoids, r2 = 0.912; p ≤ 0.05. This indicated that increased oxidative stress at high application rates of ABA, improved the production of phytochemicals.

  5. Enhancement of anti-inflammatory activity of Aloe vera adventitious root extracts through the alteration of primary and secondary metabolites via salicylic acid elicitation.

    Yun Sun Lee

    Full Text Available Aloe vera (Asphodeloideae is a medicinal plant in which useful secondary metabolites are plentiful. Among the representative secondary metabolites of Aloe vera are the anthraquinones including aloe emodin and chrysophanol, which are tricyclic aromatic quinones synthesized via a plant-specific type III polyketide biosynthesis pathway. However, it is not yet clear which cellular responses can induce the pathway, leading to production of tricyclic aromatic quinones. In this study, we examined the effect of endogenous elicitors on the type III polyketide biosynthesis pathway and identified the metabolic changes induced in elicitor-treated Aloe vera adventitious roots. Salicylic acid, methyl jasmonate, and ethephon were used to treat Aloe vera adventitious roots cultured on MS liquid media with 0.3 mg/L IBA for 35 days. Aloe emodin and chrysophanol were remarkably increased by the SA treatment, more than 10-11 and 5-13 fold as compared with untreated control, respectively. Ultra-performance liquid chromatography-electrospray ionization mass spectrometry analysis identified a total of 37 SA-induced compounds, including aloe emodin and chrysophanol, and 3 of the compounds were tentatively identified as tricyclic aromatic quinones. Transcript accumulation analysis of polyketide synthase genes and gas chromatography mass spectrometry showed that these secondary metabolic changes resulted from increased expression of octaketide synthase genes and decreases in malonyl-CoA, which is the precursor for the tricyclic aromatic quinone biosynthesis pathway. In addition, anti-inflammatory activity was enhanced in extracts of SA-treated adventitious roots. Our results suggest that SA has an important role in activation of the plant specific-type III polyketide biosynthetic pathway, and therefore that the efficacy of Aloe vera as medicinal agent can be improved through SA treatment.

  6. The Biodiversity of Lactic Acid Bacteria in Greek Traditional Wheat Sourdoughs Is Reflected in Both Composition and Metabolite Formation

    De Vuyst, Luc; Schrijvers, Vincent; Paramithiotis, Spiros; Hoste, Bart; Vancanneyt, Marc; Swings, Jean; Kalantzopoulos, George; Tsakalidou, Effie; Messens, Winy

    2002-01-01

    Lactic acid bacteria (LAB) were isolated from Greek traditional wheat sourdoughs manufactured without the addition of baker's yeast. Application of sodium dodecyl sulfate-polyacrylamide gel electrophoresis of total cell protein, randomly amplified polymorphic DNA-PCR, DNA-DNA hybridization, and 16S ribosomal DNA sequence analysis, in combination with physiological traits such as fructose fermentation and mannitol production, allowed us to classify the isolated bacteria into the species Lactob...

  7. Fungal biotransformation of chlorogenic and caffeic acids by Fusarium graminearum: New insights in the contribution of phenolic acids to resistance to deoxynivalenol accumulation in cereals.

    Gauthier, Léa; Bonnin-Verdal, Marie-Noelle; Marchegay, Gisèle; Pinson-Gadais, Laetitia; Ducos, Christine; Richard-Forget, Florence; Atanasova-Penichon, Vessela

    2016-03-16

    Fusarium Head Blight and Gibberella Ear Rot, mainly caused by the fungi Fusarium graminearum and Fusarium culmorum, are two of the most devastating diseases of small-grain cereals and maize. In addition to yield loss, these diseases frequently result in contamination of kernels with toxic type B trichothecenes. The potential involvement of chlorogenic acid in cereal resistance to Fusarium Head Blight and Gibberella Ear Rot and to trichothecene accumulation was the focus of this study. The effects of chlorogenic acid and one of its hydrolyzed products, caffeic acid, on fungal growth and type B trichothecenes biosynthesis were studied using concentrations close to physiological amounts quantified in kernels and a set of F. graminearum and F. culmorum strains. Both chlorogenic and caffeic acids negatively impact fungal growth and mycotoxin production, with caffeic acid being significantly more toxic. Inhibitory efficiencies of both phenolic acids were strain-dependent. To further investigate the antifungal and anti "mycotoxin" effect of chlorogenic and caffeic acids, the metabolic fate of these two phenolic acids was characterized in supplemented F. graminearum broths. For the first time, our results demonstrated the ability of F. graminearum to degrade chlorogenic acid into caffeic, hydroxychlorogenic and protocatechuic acids and caffeic acid into protocatechuic and hydroxycaffeic acids. Some of these metabolic products can contribute to the inhibitory efficiency of chlorogenic acid that, therefore, can be compared as a "pro-drug". As a whole, our data corroborate the contribution of chlorogenic acid to the chemical defense that cereals employ to counteract F. graminearum and its production of mycotoxins. PMID:26812586

  8. Δ9-Tetrahydrocannabinolic acid synthase: The application of a plant secondary metabolite enzyme in biocatalytic chemical synthesis.

    Lange, Kerstin; Schmid, Andreas; Julsing, Mattijs K

    2016-09-10

    Δ(9)-Tetrahydrocannabinolic acid synthase (THCAS) from the secondary metabolism of Cannabis sativa L. catalyzes the oxidative formation of an intramolecular CC bond in cannabigerolic acid (CBGA) to synthesize Δ(9)-tetrahydrocannabinolic acid (THCA), which is the direct precursor of Δ(9)-tetrahydrocannabinol (Δ(9)-THC). Aiming on a biotechnological production of cannabinoids, we investigated the potential of the heterologously produced plant oxidase in a cell-free system on preparative scale. THCAS was characterized in an aqueous/organic two-liquid phase setup in order to solubilize the hydrophobic substrate and to allow in situ product removal. Compared to the single phase aqueous setup the specific activity decreased by a factor of approximately 2 pointing to a substrate limitation of CBGA in the two-liquid phase system. However, the specific activity remained stable for at least 3h illustrating the benefit of the two-liquid phase setup. In a repeated-batch setup, THCAS showed only a minor loss of specific activity in the third batch pointing to a high intrinsic stability and high solvent tolerance of the enzyme. Maximal space-time-yields of 0.121gL(-1)h(-1) were reached proving the two-liquid phase concept suitable for biotechnological production of cannabinoids. PMID:27369551

  9. Phylogenomic evidence for a myxococcal contribution to the mitochondrial fatty acid beta-oxidation.

    Agatha Schlüter

    Full Text Available BACKGROUND: The origin of eukaryotes remains a fundamental question in evolutionary biology. Although it is clear that eukaryotic genomes are a chimeric combination of genes of eubacterial and archaebacterial ancestry, the specific ancestry of most eubacterial genes is still unknown. The growing availability of microbial genomes offers the possibility of analyzing the ancestry of eukaryotic genomes and testing previous hypotheses on their origins. METHODOLOGY/PRINCIPAL FINDINGS: Here, we have applied a phylogenomic analysis to investigate a possible contribution of the Myxococcales to the first eukaryotes. We conducted a conservative pipeline with homologous sequence searches against a genomic sampling of 40 eukaryotic and 357 prokaryotic genomes. The phylogenetic reconstruction showed that several eukaryotic proteins traced to Myxococcales. Most of these proteins were associated with mitochondrial lipid intermediate pathways, particularly enzymes generating reducing equivalents with pivotal roles in fatty acid β-oxidation metabolism. Our data suggest that myxococcal species with the ability to oxidize fatty acids transferred several genes to eubacteria that eventually gave rise to the mitochondrial ancestor. Later, the eukaryotic nucleocytoplasmic lineage acquired those metabolic genes through endosymbiotic gene transfer. CONCLUSIONS/SIGNIFICANCE: Our results support a prokaryotic origin, different from α-proteobacteria, for several mitochondrial genes. Our data reinforce a fluid prokaryotic chromosome model in which the mitochondrion appears to be an important entry point for myxococcal genes to enter eukaryotes.

  10. The short-chain fatty acid receptor, FFA2, contributes to gestational glucose homeostasis.

    Fuller, Miles; Priyadarshini, Medha; Gibbons, Sean M; Angueira, Anthony R; Brodsky, Michael; Hayes, M Geoffrey; Kovatcheva-Datchary, Petia; Bäckhed, Fredrik; Gilbert, Jack A; Lowe, William L; Layden, Brian T

    2015-11-15

    The structure of the human gastrointestinal microbiota can change during pregnancy, which may influence gestational metabolism; however, a mechanism of action remains unclear. Here we observed that in wild-type (WT) mice the relative abundance of Actinobacteria and Bacteroidetes increased during pregnancy. Along with these changes, short-chain fatty acids (SCFAs), which are mainly produced through gut microbiota fermentation, significantly changed in both the cecum and peripheral blood throughout gestation in these mice. SCFAs are recognized by G protein-coupled receptors (GPCRs) such as free fatty acid receptor-2 (FFA2), and we have previously demonstrated that the fatty acid receptor-2 gene (Ffar2) expression is higher in pancreatic islets during pregnancy. Using female Ffar2-/- mice, we explored the physiological relevance of signaling through this GPCR and found that Ffar2-deficient female mice developed fasting hyperglycemia and impaired glucose tolerance in the setting of impaired insulin secretion compared with WT mice during, but not before, pregnancy. Insulin tolerance tests were similar in Ffar2-/- and WT mice before and during pregnancy. Next, we examined the role of FFA2 in gestational β-cell mass, observing that Ffar2-/- mice had diminished gestational expansion of β-cells during pregnancy. Interestingly, mouse genotype had no significant impact on the composition of the gut microbiome, but did affect the observed SCFA profiles, suggesting a functional difference in the microbiota. Together, these results suggest a potential link between increased Ffar2 expression in islets and the alteration of circulating SCFA levels, possibly explaining how changes in the gut microbiome contribute to gestational glucose homeostasis. PMID:26394664

  11. Structural requirements for activation of the 5-oxo-6E,8Z, 11Z,14Z-eicosatetraenoic acid (5-oxo-ETE) receptor: identification of a mead acid metabolite with potent agonist activity.

    Patel, Pranav; Cossette, Chantal; Anumolu, Jaganmohan R; Gravel, Sylvie; Lesimple, Alain; Mamer, Orval A; Rokach, Joshua; Powell, William S

    2008-05-01

    The 5-lipoxygenase product 5-oxo-6E,8Z,11Z,14Z-eicosatetraenoic acid (5-oxo-ETE) is a potent chemoattractant for neutrophils and eosinophils, and its actions are mediated by the oxoeicosanoid (OXE) receptor, a member of the G protein-coupled receptor family. To define the requirements for activation of the OXE receptor, we have synthesized a series of 5-oxo-6E,8Z-dienoic acids with chain lengths between 12 and 20 carbons, as well as a series of 20-carbon 5-oxo fatty acids, either fully saturated or containing between one and five double bonds. The effects of these compounds on neutrophils (calcium mobilization, CD11b expression, and cell migration) and eosinophils (actin polymerization) were compared with those of 5-oxo-ETE. The C12 and C14 analogs were without appreciable activity, whereas the C16 5-oxo-dienoic acid was a weak partial agonist. In contrast, the corresponding C18 analog (5-oxo-18:2) was nearly as potent as 5-oxo-ETE. Among the C20 analogs, the fully saturated compound had virtually no activity, whereas 5-oxo-6E-eicosenoic acid had only weak agonist activity. In contrast, 5-oxo-6E,8Z,11Z-eicosatrienoic acid (5-oxo-20:3) and its 8-trans isomer were approximately equipotent with 5-oxo-ETE in activating granulocytes. Because of the potent effects of 5-oxo-20:3, we investigated its formation from Mead acid (5Z,8Z,11Z-eicosatrienoic acid), which accumulates in dietary essential fatty acid deficiency, by neutrophils. The main Mead acid metabolite identified was 5-hydroxy-6,8,11-eicosatrienoic acid, followed by 5-oxo-20:3 and two 6-trans isomers of leukotriene B(3). We conclude that optimal activation of the OXE receptor is achieved with 5-oxo-ETE, 5-oxo-18:2, and 5-oxo-20:3, and that the latter compound could potentially be formed under conditions of essential fatty acid deficiency. PMID:18292294

  12. Polycyclic aromatic acids are primary metabolites of alkyl-PAHs - a case study with Nereis diversicolor

    Malmquist, Linus Mattias Valdemar; Selck, Henriette; Jørgensen, Kåre Bredeli; Christensen, Jan H.

    2015-01-01

    Although concentrations of alkylated polycyclic aromatic hydrocarbons (alkyl-PAHs) in oil-contaminated sediments are higher than those of unsubstituted PAHs, only little attention has been given to metabolism and ecotoxicity of alkyl-PAHs. In this study we demonstrated that metabolism of alkyl......-PAHs primarily forms polycyclic aromatic acids (PAAs). We generalize this to other alkyl-PAHs, based on literature and the present study of the metabolism of 1-methylphenanthrene, 3,6-dimethylphenanthrene, and 1-, 2-, 3-, and 6-methylchrysene related to their unsubstituted parent PAHs. Also, we observed that...

  13. Metabolite of 15-p-iodophenyl-3(R,S)-pentadecanoic acid (123I) in blood and urine

    We analyzed matabolites of 123I- 15-(p-iodophenyl)-3(R,S)-methyl pentadecanoic acid (BMIPP) in blood and urine using rats, rabbits and human, while human samples were obtained from normal volunteers of phase 1 clinical study. We estimated metabolic pathway of 123I-BMIPP as a myocardial metabolic imaging agent. Radioactivity accumulated in heart after administration gradually decreased and was mainly excreted to bladder via kidneys. The main radioactive component in blood was 123I-p-iodophenyl acetic acid (PIPA) for any species and the urinary components were metabolic conjugates of 123I-PIPA. As results of these studies, we considered that 123I-BMIPP was metabolized to 123I-PIPA by α-oxidation process for the first step, follow by β-oxidation process, then 123I-PIPA was released to blood from tissues. Moreover, 123I-PIPA in blood was conjugated with other compounds and excreted to the bladder. (author)

  14. Contribution to the study of the oxidation reaction of Np(V) by nitric acid catalyzed par nitrous acid

    The oxidation reaction kinetics of Np(V) to Np(VI) by nitric acid catalyzed by nitrous acid was studied. In a first part, a detailed bibliographical survey was made of the oxidation-reduction reactions of U, Np, Pu, Am with nitrous and nitric acids (51 references). It is shown that only when both the organic and aqueous phases are mixed up, the extraction of a reaction product (NpVI) induces an equilibrium displacement. TBP was used as solvent. It is shown that the extraction of nitrous acid from the solvent enables the nitrous acid concentration to be kept constant and in the same order of magnitude than that of Np. This enables to show that Np(V) and nitrous acid have no simple orders. The temperature and nitric acid concentration dependence was studied. It is shown that tetravalent nitrogen must play a major part in the Np(V) oxidation

  15. Differential Contribution of Endoplasmic Reticulum and Chloroplast ω-3 Fatty Acid Desaturase Genes to the Linolenic Acid Content of Olive (Olea europaea) Fruit.

    Hernández, M Luisa; Sicardo, M Dolores; Martínez-Rivas, José M

    2016-01-01

    Linolenic acid is a polyunsaturated fatty acid present in plant lipids, which plays key roles in plant metabolism as a structural component of storage and membrane lipids, and as a precursor of signaling molecules. The synthesis of linolenic acid is catalyzed by two different ω-3 fatty acid desaturases, which correspond to microsomal- (FAD3) and chloroplast- (FAD7 and FAD8) localized enzymes. We have investigated the specific contribution of each enzyme to the linolenic acid content in olive fruit. With that aim, we isolated two different cDNA clones encoding two ω-3 fatty acid desaturases from olive (Olea europaea cv. Picual). Sequence analysis indicates that they code for microsomal (OepFAD3B) and chloroplast (OepFAD7-2) ω-3 fatty acid desaturase enzymes, different from the previously characterized OekFAD3A and OekFAD7-1 genes. Functional expression in yeast of the corresponding OepFAD3A and OepFAD3B cDNAs confirmed that they encode microsomal ω-3 fatty acid desaturases. The linolenic acid content and transcript levels of olive FAD3 and FAD7 genes were measured in different tissues of Picual and Arbequina cultivars, including mesocarp and seed during development and ripening of olive fruit. Gene expression and lipid analysis indicate that FAD3A is the gene mainly responsible for the linolenic acid present in the seed, while FAD7-1 and FAD7-2 contribute mostly to the linolenic acid present in the mesocarp and, therefore, in the olive oil. These results also indicate the relevance of lipid trafficking between the endoplasmic reticulum and chloroplast in determining the linolenic acid content of membrane and storage lipids in oil-accumulating photosynthetic tissues. PMID:26514651

  16. Influence of tannic acid application on alfalfa hay: in vitro rumen fermentation, serum metabolites and nitrogen balance in sheep.

    Getachew, G; Pittroff, W; Depeters, E J; Putnam, D H; Dandekar, A; Goyal, S

    2008-03-01

    Alfalfa protein is poorly utilised by ruminants due to its rapid degradation in rumen. The objective of the study was to assess the influence of spraying tannic acid (TA) on chopped alfalfa hay on in vitro rumen fermentation and nitrogen (N) retention by sheep. Alfalfa hay with and without TA was fed to sheep to determine nutrient digestibility and N balance. TA was sprayed on chopped alfalfa at three concentrations to determine its effect on in vitro fermentation of dry matter (DM) and N balance in sheep. Final TA concentrations were 0, 30, 60 and 90 g TA per kg DM. The control was sprayed with the same amount of water but without TA. In vitro DM degradation and the production of gas, ammonium-N (NH4-N) and short-chain fatty acid (SCFA) were measured. TA-sprayed alfalfa and the control were fed to sheep to determine nutrient digestibility and N retention. Addition of TA had no influence on the extent and rate of gas production but significantly decreased NH4-N concentration at 30 (P 0.05) after 48 h of incubation. There was a strong linear relationship between percentage increase in gas production due to PEG and protein precipitation capacity (R2 = 0.94). N digestibility was significantly reduced with all three levels of TA additions. However, the proportion of urine-N to total N output was reduced by adding 60 g (P 0.05). Higher faecal N as the TA level increased indicates incomplete dissociation of tannin-protein complexes post ruminally. Factors affecting dissociation of tannin-protein complexes need further study. PMID:22445040

  17. Volatile Metabolites

    Daryl D. Rowan

    2011-11-01

    Full Text Available Volatile organic compounds (volatiles comprise a chemically diverse class of low molecular weight organic compounds having an appreciable vapor pressure under ambient conditions. Volatiles produced by plants attract pollinators and seed dispersers, and provide defense against pests and pathogens. For insects, volatiles may act as pheromones directing social behavior or as cues for finding hosts or prey. For humans, volatiles are important as flavorants and as possible disease biomarkers. The marine environment is also a major source of halogenated and sulfur-containing volatiles which participate in the global cycling of these elements. While volatile analysis commonly measures a rather restricted set of analytes, the diverse and extreme physical properties of volatiles provide unique analytical challenges. Volatiles constitute only a small proportion of the total number of metabolites produced by living organisms, however, because of their roles as signaling molecules (semiochemicals both within and between organisms, accurately measuring and determining the roles of these compounds is crucial to an integrated understanding of living systems. This review summarizes recent developments in volatile research from a metabolomics perspective with a focus on the role of recent technical innovation in developing new areas of volatile research and expanding the range of ecological interactions which may be mediated by volatile organic metabolites.

  18. Origin of membrane dipole potential: contribution of the phospholipid fatty acid chains.

    Peterson, Uwe; Mannock, David A; Lewis, Ruthven N A H; Pohl, Peter; McElhaney, Ronald N; Pohl, Elena E

    2002-08-01

    The large intrinsic membrane dipole potential, phi(d), is important for protein insertion and functioning as well as for ion transport across natural and model membranes. However, the origin of phi(d) is controversial. From experiments carried out with lipid monolayers, a significant dependence on the fatty acid chain length is suggested, whereas in experiments with lipid bilayers, the contribution of additional -CH(2)-groups seems negligibly small compared with that of the phospholipid carbonyl groups and lipid-bound water molecules. To compare the impact of the -CH(2)-groups of dipalmitoylphosphatidylcholine (DPPC) near and far from the glycerol backbone, we have varied the structure of DPPC by incorporation of sulfur atoms in place of methylene groups in different positions of the fatty acid chain. The phi(d) of symmetric lipid bilayers containing one heteroatom was obtained from the charge relaxation of oppositely charged hydrophobic ions. We have found that the substitution for a S-atom of a -CH(2)-group decreases phi(d). The effect (deltaphi(d) = -22.6 mV) is most pronounced for S-atoms near the lipid head group while a S-atom substitution in the C(13)- or C(14)-position of the hydrocarbon chain does not effect the bilayer dipole potential. Most probably deltaphi(d) does not originate from an altered dipole potential of the acyl chain containing an heteroatom but is mediated by the disruption of chain packing, leading to a decreased density of lipid dipoles in the membrane. PMID:12191841

  19. Omega-3 Fatty Acids Protect Renal Functions by Increasing Docosahexaenoic Acid-Derived Metabolite Levels in SHR.Cg-Leprcp/NDmcr Rats, a Metabolic Syndrome Model

    Masanori Katakura

    2014-03-01

    Full Text Available The omega-3 polyunsaturated fatty acids (ω-3 PUFAs docosahexaenoic acid (DHA and/or eicosapentaenoic acid (EPA protect against diabetic nephropathy by inhibiting inflammation. The aim of this study was to assess the effects of highly purified DHA and EPA or EPA only administration on renal function and renal eicosanoid and docosanoid levels in an animal model of metabolic syndrome, SHR.Cg-Leprcp/NDmcr (SHRcp rats. Male SHRcp rats were divided into 3 groups. Control (5% arabic gum, TAK-085 (300 mg/kg/day, containing 467 mg/g EPA and 365 mg/g DHA, or EPA (300 mg/kg/day was orally administered for 20 weeks. The urinary albumin to creatinine ratio in the TAK-085-administered group was significantly lower than that in other groups. The glomerular sclerosis score in the TAK-085-administered group was significantly lower than that in the other groups. Although DHA levels were increased in total kidney fatty acids, the levels of nonesterified DHA were not significantly different among the 3 groups, whereas the levels of protectin D1, resolvin D1, and resolvin D2 were significantly increased in the TAK-085-administered group. The results show that the use of combination therapy with DHA and EPA in SHRcp rats improved or prevented renal failure associate with metabolic syndrome with decreasing triglyceride levels and increasing ω-3 PUFA lipid mediators.

  20. Evaluation of 4-(N-methylnitrosamino)-4-(3-pyridyl)butyric acid as a potential monitor of endogenous nitrosation of nicotine and its metabolites.

    Tricker, A R; Scherer, G; Conze, C; Adlkofer, F; Pachinger, A; Klus, H

    1993-07-01

    The potential endogenous nitrosation of nicotine and cotinine to yield 4-(N-methylnitrosamino)-4-(3-pyridyl)butyric acid (Iso-NNAC) has been studied in smokers and non-smokers. Following i.v. administration of 100 micrograms Iso-NNAC to rats, excretion in urine (67.4 +/- 25.4%) and feces (6.1 +/- 1.6%) occurred within 24 h. The urinary excretion of nitrate, nicotine, cotinine and Iso-NNAC were determined in 24 h urine samples from 19 smokers and 10 non-smokers. Iso-NNAC excretion was found on four occasions (44, 65, 74 and 163 ng/day) in smokers; non-smokers did not excrete Iso-NNAC. Oral administration of nicotine (n = 8; 12-40 mg) and cotinine (n = 3; 40-60 mg) to abstinent smokers did not result in Iso-NNAC excretion, even after oral nitrate (150 mg) supplementation. However, Iso-NNAC was found in cigarette tobacco (10-330 ng/g) and mainstream cigarette smoke (1.1-5.5 ng/cig.). Our studies suggest that the occasional presence of Iso-NNAC in smokers' urine results from exogenous exposure to the preformed compound in mainstream cigarette smoke and not from endogenous nitrosation of nicotine and its metabolites. PMID:8330358

  1. Effects of dietary n-3 fatty acids and vitamin C on semen characteristics, lipid composition of sperm and blood metabolites in fat-tailed Moghani rams.

    Jafaroghli, M; Abdi-Benemar, H; Zamiri, M J; Khalili, B; Farshad, A; Shadparvar, A A

    2014-06-10

    Sixteen fertile rams were randomly allotted to four groups and fed either of the four diets for 14 weeks: (1) control diet (COD) without fish oil (FO) and vitamin C (VC), (2) diet containing 2.5% FO (FOD), (3) diet containing 300 mg/kg DM VC (VCD), and (4) diet containing 2.5% FO and 300 mg/kg DM VC (FCD). Semen was collected at 14-d intervals from 1 April to 10 July (out of the physiologic breeding season in Iran). Semen volume and percentages of motile and progressively motile sperm were increased by FO and VC feeding. A significant interaction was also found between FOD and VCD on motility and progressive motility percentage (P<0.05). HOS-test and percentage of sperm with normal acrosome improved significantly by FO and VC. Rams fed FCD had better HOS-test and higher proportion of sperm with normal acrosome than rams in other groups (82.4 and 93.6%, respectively). Diets containing FO and FO and VC increased the proportion of docosahexaenoic acid in sperm (P<0.05). The activity of lactate dehydrogenase in the seminal fluid was significantly affected by VC and the interaction between FO and VC (P<0.05). Blood metabolites, except glucose, were affected positively by FO. The results showed that dietary supplementation with FO and VC improved seminal quality and may have beneficial effects on fertility in Moghani rams. PMID:24745668

  2. 3,4-Dihydroxyphenylacetic acid, a microbiota-derived metabolite of quercetin, attenuates acetaminophen (APAP)-induced liver injury through activation of Nrf-2.

    Xue, Huiting; Xie, Wenyan; Jiang, Zhihui; Wang, Meng; Wang, Jian; Zhao, Hongqiong; Zhang, Xiaoying

    2016-10-01

    1. Acetaminophen (APAP) overdose leads to severe hepatotoxicity. 3,4-dihydroxyphenylacetic acid (DOPAC) is a scarcely studied microbiota-derived metabolite of quercetin. The aim of this study was to determine the protective effect of DOPAC against APAP-induced liver injury. 2. Mice were treated intragastrically with DOPAC (10, 20 or 50 mg/kg) for 3 days before APAP (300 mg/kg) injection. APAP alone caused increase in serum aminotransferase levels and changes in hepatic histopathology. APAP also promoted oxidative stress by increasing lipid peroxidation and decreasing anti-oxidant enzyme activities. These events led to hepatocellular necrosis and reduced liver function. DOPAC increased nuclear factor erythroid 2-related factor 2 (Nrf-2) translocation to the nucleus and enhanced the expression of phase II enzymes and anti-oxidant enzymes, and thereby reduced APAP hepatotoxicity and enhanced anti-oxidant ability. 3. Our data provide evidence that DOPAC protected the liver against APAP-induced injury, which is involved in Nrf-2 activation, implying that DOPAC can be considered as a potential natural hepatoprotective agent. PMID:26931552

  3. Safety, tolerability and pharmacokinetics of 2-pyridylacetic acid, a major metabolite of betahistine, in a phase 1 dose escalation study in subjects with ADHD.

    Moorthy, Ganesh; Sallee, Floyd; Gabbita, Prasad; Zemlan, Frank; Sallans, Larry; Desai, Pankaj B

    2015-10-01

    Betahistine, a potent histamine H3 receptor antagonist, is being developed for the treatment of attention deficit hyperactivity disorder (ADHD) that manifests with symptoms such as hyperactivity, impulsivity and inattention. This study describes the pharmacokinetics of betahistine in ADHD subjects at doses higher than 50 mg. These assessments were made during a randomized, placebo-controlled, single blind, dose escalation study to determine the safety, tolerability and pharmacokinetics of once daily doses of 50 mg, 100 mg and 200 mg of betahistine in subjects with ADHD. Plasma levels of 2-pyridylacetic acid (2-PAA), a major metabolite of betahistine were quantified using a validated LC-MS/MS method and used for pharmacokinetic analysis and dose proportionality of betahistine. A linear relationship was observed in Cmax and AUC0-4 of 2-PAA with the betahistine dose (R2 0.9989 and 0.9978, respectively) and dose proportionality coefficients (β) for the power model were 0.8684 (Cmax) and 1.007 (AUC0-4). A population pharmacokinetic model with first-order absorption of betahistine and metabolism to 2-PAA, followed by a first-order elimination of 2-PAA provides estimates of clearance that underscored the linear increase in systemic exposure with dose. There were no serious adverse events reported in the study, betahistine was safe and well tolerated at all the dose levels tested. PMID:25904220

  4. Changes in the levels of abscisic acid and its metabolites in excised leaf blades of Xanthium strumarium during and after water stress

    Zeevaart, J.A.D.

    1980-10-01

    The time course of abscisic acid (ABA) accumulation during water stress and of degradation following rehydration was investigated by analyzing the levels of ABA and its metabolites phaseic acid (PA) and alkalihydrolyzable conjugated ABA in excised leaf blades of Xanthium strumarium. Initial purification was by reverse-phase, preparative, high performance liquid chromatography (HPLC) which did not require prior partitioning. ABA and PA were purified further by analytical HPLC with a ..mu..Bondapak-NH/sub 2/ column, and quantified by GLC with an electron capture detector. The ABA content of stressed leaves increased for 4 to 5 hours and then leveled off due to a balance between synthesis and degradation. Since PA accumulated at a constant rate throughout the wilting period, it was concluded that the rate of ABA synthesis decreased after the first 4 to 5 hours stress. Conjugated ABA increased at a low rate during stress. This is interpreted to indicate that free ABA was converted to the conjugated form, rather than the reverse. Following rehydration of wilted leaves, the ABA level immediately ceased increasing; it remained constant for 1 hour and then declined rapidly to the prestress level over a 2- to 3-hour period with a concomitant rise in the PA level. In contrast to the rapid disappearance of ABA after relief of stress, the high PA content of rehydrated leaves declined only slowly. The level of conjugated ABA did not change following rehydration, indicating that conjugation of ABA was irreversible. Detached Xanthium leaves that were subjected to a wilting-recovery-rewilting cycle in darkness responded to the second wilting period by formation of the same amount of ABA as accumulated after the first stress period.

  5. Contribution of coagulant and native microflora to the volatile-free fatty acid profile of an artisanal cheese

    Tavaria, Freni K.; Tavares, Tânia G.; Ferreira, A. C. Silva; Malcata, F. Xavier

    2006-01-01

    The contributions of the coagulant Cynara cardunculus and of the microflora of raw milk to the volatile-free fatty acid profile of Serra da Estrela cheese were evaluated. The experimental design included both a model system and, dual cheeses. The study in the model system showed that isovaleric acid was the predominant volatile compound after 7 d of ripening. The systems inoculated with Enterococcus faecium produced the highest amount of this volatile (ca. 135.8mg kg 1 curd), while t...

  6. Humic acids contribution to sedimentary organic matter on a shallow continental shelf (northern Adriatic Sea)

    Giani, M.; Rampazzo, F.; Berto, D.

    2010-12-01

    The shallow northern Adriatic Sea receives large river runoff, predominantly from the Po River, which is the main allochthonous source of nutrients and organic matter. The origin and quality of organic matter deposited in the sediments can influence the degradation processes and oxygen consumption in the bottom waters as well as the fate of many pollutants. Therefore the humic acids (HA) were quantified in surface and sub-surface sediments collected in an area of the north-western Adriatic platform south of Po River. HA showed to have a relevant contribution to sedimentary organic matter. HA content in sediments were positively correlated with the organic carbon concentration and negatively with redox potential and pH, particularly in sub-surface reduced sediments, suggesting their important role in the diagenetic processes taking place in anoxic conditions. Elemental composition of HA extracted from surface and sub-surface sediments showed a wide range of variation of the C org/N ratios which could be due to a mixed (terrestrial and marine) origin and/or an elevated bacteria degradation of nitrogen during diagenesis processes in sediments. The spectroscopic ratios A 2/A 4 and A 4/A 6 of HA confirmed a mixed origin with a high degree of condensation of the HA extracted from sediments.

  7. Volumetric behaviour of amino acids and their group contributions in aqueous lactose solutions at different temperatures

    Densities, ρ, for glycine, L-alanine, L-valine, and L-leucine [(0.05 to 0.30) m] in aqueous lactose solutions ranging from pure water to 6 mass% lactose were determined at T = (293.15, 298.15, 303.15, and 308.15) K. The density was used to compute apparent molar volume, Vφ, partial molar volume at infinite dilution, Vφo, and experimental slope, SV were obtained and interpreted in terms of solute-solvent and solute-solute interactions. These data were used to calculate the (∂Vφ0/∂T)P values. The partial molar volume of transfer, ΔVφ0 from water to aqueous lactose solutions at infinite dilution has also been calculated. In addition to this, the linear correlation of Vφ0 with number of carbon atoms in the alkyl chain of amino acids was utilized to determine the respective contributions of NH3+COO-, and CH2 groups to Vφ0.

  8. Volumetric behaviour of amino acids and their group contributions in aqueous lactose solutions at different temperatures

    Pal, Amalendu, E-mail: palchem@sify.co [Department of Chemistry, Kurukshetra University, Kurukshetra 136 119 (India); Chauhan, Nalin [Department of Chemistry, Kurukshetra University, Kurukshetra 136 119 (India)

    2011-02-15

    Densities, {rho}, for glycine, L-alanine, L-valine, and L-leucine [(0.05 to 0.30) m] in aqueous lactose solutions ranging from pure water to 6 mass% lactose were determined at T = (293.15, 298.15, 303.15, and 308.15) K. The density was used to compute apparent molar volume, V{sub {phi}}, partial molar volume at infinite dilution, V{sub {phi}}{sup o}, and experimental slope, S{sub V} were obtained and interpreted in terms of solute-solvent and solute-solute interactions. These data were used to calculate the ({partial_derivative}V{sub {phi}}{sup 0}/{partial_derivative}T){sub P} values. The partial molar volume of transfer, {Delta}V{sub {phi}}{sup 0} from water to aqueous lactose solutions at infinite dilution has also been calculated. In addition to this, the linear correlation of V{sub {phi}}{sup 0} with number of carbon atoms in the alkyl chain of amino acids was utilized to determine the respective contributions of NH{sub 3}{sup +}COO{sup -}, and CH{sub 2} groups to V{sub {phi}}{sup 0}.

  9. Carbonyl reductase inactivation may contribute to mouse lung tumor promotion by electrophilic metabolites of butylated hydroxytoluene: protein alkylation in vivo and in vitro.

    Shearn, Colin T; Fritz, Kristofer S; Meier, Brent W; Kirichenko, Oleg V; Thompson, John A

    2008-08-01

    Promotion of lung tumors in mice by the food additive butylated hydroxytoluene (BHT) is mediated by electrophilic metabolites produced in the target organ. Identifying the proteins alkylated by these quinone methides (QMs) is a necessary step in understanding the underlying mechanisms. Covalent adducts of the antioxidant enzymes peroxiredoxin 6 and Cu,Zn superoxide dismutase were detected previously in lung cytosols from BALB/c mice injected with BHT, and complimentary in vitro studies demonstrated that QM alkylation causes inactivation and enhances oxidative stress. In the present work, adducts of another protective enzyme, carbonyl reductase (CBR), were detected by Western blotting and mass spectrometry in mitochondria from lungs of mice one day after a single injection of BHT and throughout a 28-day period of weekly injections required to achieve tumor promotion. BHT treatment was accompanied by the accumulation of protein carbonyls in lung cytosol from sustained oxidative stress. Studies in vitro demonstrated that CBR activity in lung homogenates was susceptible to concentration- and time-dependent inhibition by QMs. Recombinant CBR underwent irreversible inhibition during QM exposure, and mass spectrometry was utilized to identify alkylation sites at Cys 51, Lys 17, Lys 189, Lys 201, His 28, and His 204. Except for Lys 17, all of these adducts were eliminated as a cause of enzyme inhibition either by chemical modification (cysteine) or site-directed mutagenesis (lysines and histidines). The data demonstrated that Lys 17 is the critical alkylation target, consistent with the role of this basic residue in NADPH binding. These data support the possibility that CBR inhibition occurs in BHT-treated mice, thereby compromising one pathway for inactivating lipid peroxidation products, particularly 4-oxo-2-nonenal. These data, in concert with previous evidence for the inactivation of antioxidant enzymes, provide a molecular basis to explain lung inflammation leading to

  10. Associations of prepartum plasma cortisol, haptoglobin, fecal cortisol metabolites, and nonesterified fatty acids with postpartum health status in Holstein dairy cows.

    Huzzey, J M; Nydam, D V; Grant, R J; Overton, T R

    2011-12-01

    The association between negative energy balance and health has led to the testing of blood analytes such as nonesterified fatty acids (NEFA) to identify opportunities for improving the management of transition dairy cows. The objective of this study was to evaluate whether prepartum analytes associated with stress (cortisol) or inflammation (haptoglobin) could also identify dairy cattle at increased risk for health complications after calving. Prepartum blood and fecal samples were collected once weekly from 412 Holstein dairy cows on 2 commercial dairy farms (at wk -3, -2, and -1 relative to calving) and analyzed for concentrations of NEFA, haptoglobin (Hp), and cortisol in plasma and cortisol metabolites in feces. Retained placenta (RP), displaced abomasum (DA), subclinical ketosis (SCK), high Hp concentration (HiHp), and death were recorded up to 30 d in milk (DIM), and animals were subsequently categorized into 3 health categories: (1) no disorder of interest (NDI); (2) one disorder (RP, DA, SCK, or HiHp); or (3) more than one disorder (RP, DA, SCK, HiHp) or death. With the exception of prepartum NEFA, no associations were detected between prepartum concentrations of our analytes of interest and the occurrence of one disorder (RP, DA, SCK, or HiHP) by 30 DIM. Haptoglobin concentration tended to be greater during wk -2 and -1 in cows that developed more than one disorder or that died by 30 DIM; however, when calving assistance was included as a covariate in the analysis prepartum, Hp was no longer a significant risk factor for this postpartum health outcome. Primiparous cows with plasma cortisol concentrations >22.2 nmol/L during wk -2 had reduced odds [odds ratio (OR) 0.41; 95% confidence interval (CI) 0.17-0.98] of developing more than one disorder or death by 30 DIM, whereas multiparous cows with plasma cortisol >34.1 nmol/L during wk -2 tended to have greater odds (OR 2.53; 95% CI 0.87-7.37) of developing more than one disorder or death by 30 DIM. Individual

  11. Maternal phenylketonuria: Embryotoxicity in vitro of PKU-related metabolites and of human PKU-sera

    Piersma AH; Verhoef A; Hamers AM; van den Ham WA; Jansen EHJM

    1993-01-01

    Mothers with untreated phenylketonuria (PKU) have an increased risk of bearing children with congenital malformations. PKU causes accumulation of phenylalanine (PHE) and its metabolites in urine and blood, and this condition may contribute to the developmental problems. In the present study we investigated the embryotoxicity of these compounds in the rat postimplantation embryo culture system. Whereas PHE was ineffective, phenylpyruvic acid and phenylacetic acid induced general growth retarda...

  12. The effects of dietary omega fatty acids on pregnancy rate, plasma prostaglandin metabolite levels, serum progesterone levels, and milk fatty-acid profile in beef cows

    Richardson, Gavin F.; McNiven, Mary A.; Petit, Hélène V.; Duynisveld, John L.

    2013-01-01

    The objectives were to determine the effects of feeding supplements rich in omega-6 or omega-3 fatty acids (FA) during the late gestation to the early postpartum and breeding periods on reproduction and milk FA profile in beef cows. For each of two years, at the beginning of period 1 (mid-December), 72 beef cows, calving in January or February, were assigned to diets supplemented with roasted flaxseed (Flax) or roasted soybean (Soybean). For each of two years, after 11 wk (end of period 1), 1...

  13. Constitutive androstane receptor-mediated changes in bile acid composition contributes to hepatoprotection from lithocholic acid-induced liver injury in mice.

    Beilke, Lisa D; Aleksunes, Lauren M; Holland, Ricky D; Besselsen, David G; Beger, Rick D; Klaassen, Curtis D; Cherrington, Nathan J

    2009-05-01

    Pharmacological activation of the constitutive androstane receptor (CAR) protects the liver during cholestasis. The current study evaluates how activation of CAR influences genes involved in bile acid biosynthesis as a mechanism of hepatoprotection during bile acid-induced liver injury. CAR activators phenobarbital (PB) and 1,4-bis[2-(3,5-dichloropyridyloxy)]benzene (TCPOBOP) or corn oil (CO) were administered to C57BL/6 wild-type (WT) and CAR knockout (CAR-null) mice before and during induction of intrahepatic cholestasis using the secondary bile acid, lithocholic acid (LCA). In LCA-treated WT and all the CAR-null groups (excluding controls), histology revealed severe multifocal necrosis. This pathology was absent in WT mice pretreated with PB and TCPOBOP, indicating CAR-dependent hepatoprotection. Decreases in total hepatic bile acids and hepatic monohydroxy, dihydroxy, and trihydroxy bile acids in PB- and TCPOBOP-pretreated WT mice correlated with hepatoprotection. In comparison, concentrations of monohydroxylated and dihydroxylated bile acids were increased in all the treated CAR-null mice compared with CO controls. Along with several other enzymes (Cyp7b1, Cyp27a1, Cyp39a1), Cyp8b1 expression was increased in hepatoprotected mice, which could be suggestive of a shift in the bile acid biosynthesis pathway toward the formation of less toxic bile acids. In CAR-null mice, these changes in gene expression were not different among treatment groups. These results suggest CAR mediates a shift in bile acid biosynthesis toward the formation of less toxic bile acids, as well as a decrease in hepatic bile acid concentrations. We propose that these combined CAR-mediated effects may contribute to the hepatoprotection observed during LCA-induced liver injury. PMID:19196849

  14. Groundwater contribution to an acid upland lake (Loch Fleet, Scotland) and the possibilities for amelioration

    Cook, J. M.; Edmunds, W. M.; Robins, N. S.

    1991-06-01

    The Loch Fleet catchment lies in an upland region in the centre of the outcrop of the Cairnsmore of Fleet granite. It is a recently acidified lake (pH = 4.4) which has been the subject of a liming experiment to restore fisheries. In the present study, hydrogeological and geochemical techniques were used to determine the contribution of ground water to the loch and its role in buffering the lake water chemistry. Diffuse groundwater seepage was detected by infrared linescan survey, and overflowing ground water (2 m above the level of the loch) was encountered in a shallow borehole. This ground water has an alkaline geochemistry (pH = 7.2, HCO 3- = 142 mg l -1) determined by secondary vein calcite and hydrolysis of silicate minerals. The net gains or losses of various constituents in the ground water and in the loch outflow have been determined relative to rainfall inputs. Na, K, Ca, Mg, HCO 3, SO 4, Cl, Si, Sr, Fe, Mn, Li and F all show net gain in the ground water; NO 3, Al, Zn and B show a net loss. In the acidic loch outflow, Ca, Mg, Si, Sr, Ba, Fe, Mn, Al, Zn and Li show a net gain over rainfall inputs; most of these elements derive from ground water, enhanced by evapotranspiration by a factor of 1.8. The chemical results have been used to determine that ground water contributes around 3.5 l s -1 to the loch, compared with an estimated 3-4 l s -1 derived from hydrograph analysis. This constitutes 5% of the mean loch outflow, which was sufficient to buffer the loch at around pH = 6.0 until the late 1960s. Titrations of ground water with loch water show that as little as 0.06 l s -1 (1656 m 3 year -1) of additional ground water would be required to restore the loch to conditions suitable for a self-sustaining fish population. Twice this flux (3310 m 3 year -1) would restore the loch to the conditions pertaining in the pre-industrial era. These targets could be achieved at an economic cost, it is suggested, by induced abstraction of ground water in the upper reaches

  15. Neuroprotective effects of vinpocetine and its major metabolite cis-apovincaminic acid on NMDA-induced neurotoxicity in a rat entorhinal cortex lesion model.

    Nyakas, Csaba; Felszeghy, Klára; Szabó, Róbert; Keijser, Jan N; Luiten, Paul G M; Szombathelyi, Zsolt; Tihanyi, Károly

    2009-01-01

    Vinpocetine (ethyl-apovincaminate, Cavinton), a synthetic derivative of the Vinca minor alkaloid vincamine, has been used now for decades for prevention and treatment of cerebrovascular diseases predisposing to development of dementia. Both vinpocetine and its main metabolite cis-apovincaminic acid (cAVA) exert a neuroprotective type of action. Bilateral N-methyl-D-aspartate (NMDA)-induced neurodegeneration in the entorhinal cortex of rat was used as a dementia model to confirm the neuroprotective action of these compounds in vivo. NMDA-lesioned rats were treated 60 min before lesion and throughout 3 postoperative days with a 10 mg/kg intraperitoneal dose of vinpocetine or cAVA. Behavioral tests started after termination of drug treatment and consisted of novel object recognition, social discrimination, and spontaneous alternation in a Y-maze, and spatial learning in the Morris water maze. At the end of behavioral testing brains were perfused with fixative and the size of the excitotoxic neuronal lesion and that of microglial activation around the lesion were assayed quantitatively on brain sections immunostained for neuron-specific nuclear protein (NeuN) and integrin CD11b, respectively. Entorhinal NMDA lesions impaired recognition of novel objects and the new social partner, and suppressed spontaneous alternation and spatial learning performance in the Morris maze. Both vinpocetine and cAVA effectively attenuated the behavioral deficits, and significantly decreased lesion size and the region of microglia activation. Both lesion-induced attention deficit and learning disabilities were markedly alleviated by vinpocetine and cAVA. The morphological findings corroborated the behavioral observations and indicated reduced lesion size and microglia activation especially after vinpocetine treatment which supports an in vivo neuroprotective mode of action of vinpocitine and a less potent action of cAVA. PMID:19492990

  16. The simultaneous identification of metoprolol and its major acidic and basic metabolites in human urine by gas chromatography-mass spectrometry

    Li, Feng; Cooper, S.F. [Universite du Quebec, Pointe-Claire (Canada)

    1996-12-31

    A novel gas chromatography-mass spectrometric (GC-MS) method was developed to confirm and identify metoprolol and its metabolites by double derivatization with S-(-)menthyl chloroformate [(-)-MCF] and N-methyl(trimethylsilyl-trifluoroacetamide) (MSTFA). This is the first report, which describes the simultaneous identification of metoprolol, its one major acidc and other basic metabolites in human urine based on solid-phase extraction with C{sub 18} reversed-phase cartridges. 12 refs., 4 figs.

  17. Contributions of separate reactions to the acid-base buffering of soils in brook floodplains (Central Forest State Reserve)

    Sokolova, T. A.; Tolpeshta, I. I.; Rusakova, E. S.

    2016-04-01

    The acid-base buffering of gleyic gray-humus soils developed in brook floodplains and undisturbed southern-taiga landscapes has been characterized by the continuous potentiometric titration of soil water suspensions. During the interaction with an acid, the major amount of protons (>80%) is consumed for the displacement of exchangeable bases and the dissolution of Ca oxalates. In the O and AY horizons, Mn compounds make the major contribution (2-15%) to the acid buffering. The buffer reactions with the participation of Al compounds make up from 0.5 to 1-2% of the total buffering capacity, and the protonation of the surface OH groups of kaolinite consumes 2-3% of the total buffering capacity. The deprotonation of OH groups on the surface of Fe hydroxides (9-43%), the deprotonation of OH groups on the surface of illite crystals (3-19%), and the dissolution of unidentified aluminosilicates (9-14%) are the most significant buffer reactions whose contributions have been quantified during the interaction with a base. The contribution of the deprotonation of OH groups on the surface of kaolinite particles is lower (1-5%) because of the small specific surface area of this mineral, and that of the dissolution of Fe compounds is insignificant. In the AY horizon, the acid and base buffering of soil in the rhizosphere is higher than beyond the rhizosphere because of the higher contents of organic matter and nonsilicate Fe and Al compounds.

  18. Nontargeted metabolite profiles and sensory properties of strawberry cultivars grown both organically and conventionally.

    Kårlund, Anna; Hanhineva, Kati; Lehtonen, Marko; Karjalainen, Reijo O; Sandell, Mari

    2015-01-28

    Strawberry (Fragaria × ananassa Duch.) contains many secondary metabolites potentially beneficial for human health, and several of these compounds contribute to strawberry sensory properties, as well. In this study, three strawberry cultivars grown both conventionally and organically were subjected to nontargeted metabolite profiling analysis with LC-qTOF-ESI-MS and to descriptive sensory evaluation by a trained panel. Combined metabolome and sensory data (PLS model) revealed that 79% variation in the metabolome explained 88% variation in the sensory profiles. Flavonoids and condensed and hydrolyzable tannins determined the orosensory properties, and fatty acids contributed to the odor attributes of strawberry. Overall, the results indicated that the chemical composition and sensory quality of strawberries grown in different cultivation systems vary mostly according to cultivar. Organic farming practices may enhance the accumulation of some plant metabolites in specific strawberry genotypes. Careful cultivar selection is a key factor for the improvement of nutritional quality and marketing value of organic strawberries. PMID:25569122

  19. Corrosion rate of construction materials in hot phosphoric acid with the contribution of anodic polarization

    Kouril, M.; Christensen, Erik; Eriksen, S.;

    2011-01-01

    The paper is focused on selection of a proper material for construction elements of water electrolysers, which make use of a 85% phosphoric acid as an electrolyte at temperature of 150 8C and which might be loaded with anodic polarization up to 2.5 V versus a saturated Ag/AgCl electrode (SSCE......% phosphoric acid at 150 8C and at polarization of 2.5 V/SSCE is tantalum. In that case, even a gentle cathodic polarization is harmful in such an acidic environment. Hydrogen reduction leads to tantalum hydride formation, to loss of mechanical properties and to complete disintegration of the metal. Contrary...... to tantalum, titanium is free of any corrosion resistance in hot phosphoric acid. Its corrosion rate ranges from tens of millimetres to metres per year depending on temperature of the acid. Alloy bonded tantalum coating was recognized as an effective corrosion protection for both titanium and stainless steel...

  20. Microbial contributions to C and N dynamics in decaying litter elucidated by amino acid and amino sugar analyses

    Hobara, S.; Osono, T.; Noro, K.; Hirota, M.; Benner, R. H.

    2011-12-01

    There is still much to be revealed about carbon (C) and nitrogen (N) dynamics in terrestrial soil systems. The objectives of this study were to identify molecular changes in composition during plant litter decomposition and gain insights about microbial contributions to C and N dynamics in decaying litter. Litter bag experiments with three plant species, Miscanthus sinensis, Pinus densiflora and Quercus crispula, were conducted for three years, and the concentrations of C, N, amino acids and amino sugars were determined at various times during the experiments. Mass loss (AFDW) ranged from 66-90% for the plant tissues. The weight %C remained fairly constant, whereas the weight %N increased throughout the study indicating N immobilization was occurring. The percentages of C as amino acids and amino sugars also increased throughout the study suggesting these biomolecules were largely of microbial origin. The increasing yields of amino acids and amino sugars were inversely related to overall C loss from the litter material. As microorganisms degraded the plant litter they left behind molecular signatures that were useful predictors of the extent of overall degradation. The C/N ratio of litter decreased throughout the study and was inversely related to galactosamine yields. The glucosamine/galactosamine (GlcN/GalN) ratio gradually declined to values near 2 by the end of the study. Galactoasamine is more abundant in bacteria than fungi, and the declining GlcN/GalN ratio suggest the relative contributions of bacterial to litter C and N increased relative to contributions from fungi. A cluster analysis of 0- and 36-month litters based on amino acid and amino sugar composition showed that 0-month litters of three plant species were separated from 36-month litters, suggesting common diagenetic pathways during decomposition irrespective of plant species. The microbial decomposers contribute to N immobilization and their contributions to the C and N content of litter increases

  1. Heterogeneous catalysis contribution for the denitration of aqueous nuclear radioactive waste with formic acid

    The chemical denitration aims to reduce the nitric acid concentration in nuclear fuel reprocessing aqueous wastes by adding formic acid as a reducing agent. The denitration reaction exhibits an induction period, which duration is related to the time needed by the key intermediate of the reaction, i.e. nitrous acid, to reach a threshold concentration in the reaction medium. The addition of a Pt/SiO2 catalyst in the reaction mixture suppresses the induction period of the chemical denitration. The aim of the present work is to identify the role of Pt/SiO2 in the catalytic denitration mechanism. The experimental work is based on the comparison of catalytic tests performed with various catalysts, in order to identify the intrinsic characteristics of Pt/SiO2 that might influence its activity for the reaction. Catalytic denitration results show that Pt/SiO2 acts only by speeding up the nitrous acid generation in the solution until its concentration reaches the threshold level of 0,01 mol L-1 in the experimental conditions. Catalysts activity is evaluated by quantifying the nitrous acid generated on the platinum surface during the induction period of the homogeneous denitration reaction. The large platinum aggregates reactivity is greater than the one of nano-sized particles. The study of the key step of the catalytic denitration reaction, the catalytic generation of nitrous acid, clarifies the role of Pt/SiO2. The homogeneous denitration is initiated thanks to a redox cycle on the catalyst surface: an initial oxidation of Pt0 by nitric acid, which is reduced into nitrous acid, followed by the reduction of the passivated 'Ptox' by formic acid. Furthermore, a platinum reduction by formic acid prior to the catalytic test prevents any platinum leaching from the catalyst into the nitric solution, being all the more significant as platinum dispersion is high. (author)

  2. Contribution to the study of pertechnetate (sup(99m)Tc) stannous citrate - citric acid complexation

    Pertechnetate/citric acid/stannous citrate complexation carried out from a lyophilisate of stannous citrate in citric medium at pH5 leads to the formation of separable compounds. These compounds are tin-free technetium citrates. Similar results have been described in the case of complexation reactions with glycolic, thioglycolic and thiomalic acids and with other carboxylates such as dimercaptosuccinic acid. These processes include the reduction of Tcsup(VIII) by Snsup(II) in the presence of thiomalic acid under conditions similar to our own: stannous thiomalate in thiomalic medium to which is added the pertechnetate solution producing Tc-thiomalate complexes variable with the reaction pH. Also worth considering is the possible complexation between pertechnetate and the same acid in the absence of reducing ion, following a special procedure (heating). The complexes described here contain the oxotechnetium bond (terminal oxygen-technetium) and a strong probability exists in favour of dimerisation. Their stability, for a reaction in acid solution: pH 5.0/5.5, becomes satisfactory if: the solution is concentrated enough; bubbling by an inert gas is carried out; room temperature is not exceeded. The development takes place through a partial reoxidation characterised by colour change. An original interaction between reduced states of Tc and citric acid may be claimed with certainty under our experimental conditions. The difficulty then lies in the passage to the tracer stage when the isotope sup(99m)Tc is used

  3. Effect of storage time on metabolite profile and alpha-glucosidase inhibitory activity of Cosmos caudatus leaves – GCMS based metabolomics approach

    Neda Javadi

    2015-09-01

    Full Text Available Cosmos caudatus, which is a commonly consumed vegetable in Malaysia, is locally known as “Ulam Raja”. It is a local Malaysian herb traditionally used as a food and medicinal herb to treat several maladies. Its bioactive or nutritional constituents consist of a wide range of metabolites, including glucosinolates, phenolics, amino acids, organic acids, and sugars. However, many of these metabolites are not stable and easily degraded or modified during storage. In order to investigate the metabolomics changes occurring during post-harvest storage, C. caudatus samples were subjected to seven different storage times (0 hours, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours, and 12 hours at room temperature. As the model experiment, the metabolites identified by gas chromatography-mass spectrometry (GC-MS were correlated with α-glucosidase inhibitory activity analyzed with multivariate data analysis (MVDA to find out the variation among samples and metabolites contributing to the activity. Orthogonal partial least squares (OPLS analysis was applied to investigate the metabolomics changes. A profound chemical alteration, both in primary and secondary metabolites, was observed. The α-tocopherol, catechin, cyclohexen-1-carboxylic acid, benzoic acid, myo-inositol, stigmasterol, and lycopene compounds were found to be the discriminating metabolites at early storage; however, sugars such as sucrose, α-d-galactopyranose, and turanose were detected, which was attributed to the discriminating metabolites for late storage. The result shows that the MVDA method is a promising technique to identify biomarker compounds relative to storage at different times.

  4. Fatty Acids Composition of Vegetable Oils and Its Contribution to Dietary Energy Intake and Dependence of Cardiovascular Mortality on Dietary Intake of Fatty Acids.

    Orsavova, Jana; Misurcova, Ladislava; Ambrozova, Jarmila Vavra; Vicha, Robert; Mlcek, Jiri

    2015-01-01

    Characterizations of fatty acids composition in % of total methylester of fatty acids (FAMEs) of fourteen vegetable oils--safflower, grape, silybum marianum, hemp, sunflower, wheat germ, pumpkin seed, sesame, rice bran, almond, rapeseed, peanut, olive, and coconut oil--were obtained by using gas chromatography (GC). Saturated (SFA), monounsaturated (MUFA) and polyunsaturated fatty acids (PUFA), palmitic acid (C16:0; 4.6%-20.0%), oleic acid (C18:1; 6.2%-71.1%) and linoleic acid (C18:2; 1.6%-79%), respectively, were found predominant. The nutritional aspect of analyzed oils was evaluated by determination of the energy contribution of SFAs (19.4%-695.7% E(RDI)), PUFAs (10.6%-786.8% E(RDI)), n-3 FAs (4.4%-117.1% E(RDI)) and n-6 FAs (1.8%-959.2% E(RDI)), expressed in % E(RDI) of 1 g oil to energy recommended dietary intakes (E(RDI)) for total fat (E(RDI)--37.7 kJ/g). The significant relationship between the reported data of total fat, SFAs, MUFAs and PUFAs intakes (% E(RDI)) for adults and mortality caused by coronary heart diseases (CHD) and cardiovascular diseases (CVD) in twelve countries has not been confirmed by Spearman's correlations. PMID:26057750

  5. Fatty Acids Composition of Vegetable Oils and Its Contribution to Dietary Energy Intake and Dependence of Cardiovascular Mortality on Dietary Intake of Fatty Acids

    Jana Orsavova

    2015-06-01

    Full Text Available Characterizations of fatty acids composition in % of total methylester of fatty acids (FAMEs of fourteen vegetable oils—safflower, grape, silybum marianum, hemp, sunflower, wheat germ, pumpkin seed, sesame, rice bran, almond, rapeseed, peanut, olive, and coconut oil—were obtained by using gas chromatography (GC. Saturated (SFA, monounsaturated (MUFA and polyunsaturated fatty acids (PUFA, palmitic acid (C16:0; 4.6%–20.0%, oleic acid (C18:1; 6.2%–71.1% and linoleic acid (C18:2; 1.6%–79%, respectively, were found predominant. The nutritional aspect of analyzed oils was evaluated by determination of the energy contribution of SFAs (19.4%–695.7% ERDI, PUFAs (10.6%–786.8% ERDI, n-3 FAs (4.4%–117.1% ERDI and n-6 FAs (1.8%–959.2% ERDI, expressed in % ERDI of 1 g oil to energy recommended dietary intakes (ERDI for total fat (ERDI—37.7 kJ/g. The significant relationship between the reported data of total fat, SFAs, MUFAs and PUFAs intakes (% ERDI for adults and mortality caused by coronary heart diseases (CHD and cardiovascular diseases (CVD in twelve countries has not been confirmed by Spearman’s correlations.

  6. Eicosapentaenoic Acid Inhibits Oxidation of ApoB-containing Lipoprotein Particles of Different Size In Vitro When Administered Alone or in Combination With Atorvastatin Active Metabolite Compared With Other Triglyceride-lowering Agents.

    Mason, R Preston; Sherratt, Samuel C R; Jacob, Robert F

    2016-07-01

    Eicosapentaenoic acid (EPA) is a triglyceride-lowering agent that reduces circulating levels of the apolipoprotein B (apoB)-containing lipoprotein particles small dense low-density lipoprotein (sdLDL), very-low-density lipoprotein (VLDL), and oxidized low-density lipoprotein (LDL). These benefits may result from the direct antioxidant effects of EPA. To investigate this potential mechanism, these particles were isolated from human plasma, preincubated with EPA in the absence or presence of atorvastatin (active) metabolite, and subjected to copper-initiated oxidation. Lipid oxidation was measured as a function of thiobarbituric acid reactive substances formation. EPA inhibited sdLDL (IC50 ∼2.0 μM) and LDL oxidation (IC50 ∼2.5 μM) in a dose-dependent manner. Greater antioxidant potency was observed for EPA in VLDL. EPA inhibition was enhanced when combined with atorvastatin metabolite at low equimolar concentrations. Other triglyceride-lowering agents (fenofibrate, niacin, and gemfibrozil) and vitamin E did not significantly affect sdLDL, LDL, or VLDL oxidation compared with vehicle-treated controls. Docosahexaenoic acid was also found to inhibit oxidation in these particles but over a shorter time period than EPA. These data support recent clinical findings and suggest that EPA has direct antioxidant benefits in various apoB-containing subfractions that are more pronounced than those of other triglyceride-lowering agents and docosahexaenoic acid. PMID:26945158

  7. Online restricted-access material combined with high-performance liquid chromatography and tandem mass spectrometry for the simultaneous determination of vanillin and its vanillic acid metabolite in human plasma.

    Li, De-Qiang; Zhang, Zhi-Qing; Yang, Xiu-Ling; Zhou, Chun-Hua; Qi, Jin-Long

    2016-09-01

    An automated online solid-phase extraction with restricted-access material combined with high-performance liquid chromatography and tandem mass spectrometry was developed and validated for the simultaneous quantification of vanillin and its vanillic acid metabolite in human plasma. After protein precipitation by methanol, which contained the internal standards, the supernatant of plasma samples was injected to the system, the endogenous large molecules were flushed out, and target analytes were trapped and enriched on the adsorbent, resulting in a minimization of sample complexity and ion suppression effects. Calibration curves were linear over the concentrations of 5-1000 ng/mL for vanillin and 10-5000 ng/mL for vanillic acid with a coefficient of determination >0.999 for the determined compounds. The lower limits of quantification of vanillin and vanillic acid were 5.0 and 10.0 ng/mL, respectively. The intra- and inter-run precisions expressed as the relative standard deviation were 2.6-8.6 and 3.2-10.2%, respectively, and the accuracies expressed as the relative error were in the range of -6.1 to 7.3%. Extraction recoveries of analytes were between 89.5 and 97.4%. There was no notable matrix effect for any analyte concentration. The developed method was proved to be sensitive, repeatable, and accurate for the quantification of vanillin and its vanillic acid metabolite in human plasma. PMID:27384745

  8. Essential fatty acids and their metabolites could function as endogenous HMG-CoA reductase and ACE enzyme inhibitors, anti-arrhythmic, anti-hypertensive, anti-atherosclerotic, anti-inflammatory, cytoprotective, and cardioprotective molecules

    Das Undurti N

    2008-10-01

    Full Text Available Abstract Lowering plasma low density lipoprotein-cholesterol (LDL-C, blood pressure, homocysteine, and preventing platelet aggregation using a combination of a statin, three blood pressure lowering drugs such as a thiazide, a β blocker, and an angiotensin converting enzyme (ACE inhibitor each at half standard dose; folic acid; and aspirin-called as polypill- was estimated to reduce cardiovascular events by ~80%. Essential fatty acids (EFAs and their long-chain metabolites: γ-linolenic acid (GLA, dihomo-GLA (DGLA, arachidonic acid, eicosapentaenoic acid (EPA, and docosahexaenoic acid (DHA and other products such as prostaglandins E1 (PGE1, prostacyclin (PGI2, PGI3, lipoxins (LXs, resolvins, protectins including neuroprotectin D1 (NPD1 prevent platelet aggregation, lower blood pressure, have anti-arrhythmic action, reduce LDL-C, ameliorate the adverse actions of homocysteine, show anti-inflammatory actions, activate telomerase, and have cytoprotective properties. Thus, EFAs and their metabolites show all the classic actions expected of the "polypill". Unlike the proposed "polypill", EFAs are endogenous molecules present in almost all tissues, have no significant or few side effects, can be taken orally for long periods of time even by pregnant women, lactating mothers, and infants, children, and adults; and have been known to reduce the incidence cardiovascular diseases including stroke. In addition, various EFAs and their long-chain metabolites not only enhance nitric oxide generation but also react with nitric oxide to yield their respective nitroalkene derivatives that produce vascular relaxation, inhibit neutrophil degranulation and superoxide formation, inhibit platelet activation, and possess PPAR-γ ligand activity and release NO, thus prevent platelet aggregation, thrombus formation, atherosclerosis, and cardiovascular diseases. Based on these evidences, I propose that a rational combination of ω-3 and ω-6 fatty acids and the co

  9. Characterization of humic acids, the Riso contribution to the Coco-club activities

    Within the framework of the CEC project Mirage 2/Coco Club, a series of humic acids has been characterized by moisture content, ash content, metal content, functional group content and complexional behaviour towards europium ions

  10. Contribution au profilage des acides organiques urinaires, chez l'enfant

    Pérez-Vásquez, Naira

    2015-01-01

    Among inherited metabolic diseases, organic acidemia (OA) or organic aciduria is characterized by urinary excretion of abnormal amounts or types of organic acids. OA is mostly associated with genetic conditions resulting in a specific step of amino acid catabolism dysfunction. Such alterations can produce disease states that range from mild to lethal neurological involvement. Gas chromatography coupled to mass spectrometry (GC-MS) remains the most used analytical technique for detecting speci...

  11. Acid rain research[NIVA contributions to ACID REIGN '95? Conference in Gothenburg, Sweden, 26-30 June 1995

    NONE

    1995-07-01

    The conference dealt with various sources, aspects and present and future consequences of pollution, particularly acid precipitation on the environment especially on the terrestrial and aquatic parts, with the focus on acidification. Surveys of the extent of the problems were presented. Various methods of reversing the effects are dealt with. Much attention was focused on various water systems such as rivers, lakes, surface waters and ground water and consequences of pollution chemically, geologically, biologically and botanically. The problems of global warming and changes were also focused upon. The origins of pollutants were discussed.

  12. Contribution of several amino acids and lactate to gluconeogenesis in hepatocytes isolated from rats fed various diets

    The contribution under various nutritional regimens of several amino acids and lactate to gluconeogenesis was estimated by measuring the glucose formation from 14C-labeled substrates. Isolated rat hepatocytes were incubated for 60 min in a Krebs-Ringer bicarbonate buffer pH 7.4 containing lactate, pyruvate and all the amino acids at concentrations similar to their physiological levels found in rat plasma, with one precursor labeled in each flask. In all conditions, lactate was the major glucose precursor, providing over 60% of the glucose formed. Glutamine and alanine were the major amino acid precursors of glucose, contributing 9.8% and 10.6% of the glucose formed, respectively, in hepatocytes isolated from starved rats. Serine, glycine and threonine also contributed to gluconeogenesis in the starved liver cells at 2.6, 2.1 and 3.8%, respectively, of the glucose formed. The rate of glucose formation from the isolated hepatocytes of the starved rats and those fed either high protein or high fat was higher than that from rats fed a nonpurified diet

  13. Increased hepatic Fatty Acid uptake and esterification contribute to tetracycline-induced steatosis in mice.

    Choi, You-Jin; Lee, Chae-Hyeon; Lee, Kang-Yo; Jung, Seung-Hwan; Lee, Byung-Hoon

    2015-06-01

    Tetracycline induces microvesicular steatosis, which has a poor long-term prognosis and a higher risk of steatohepatitis development compared with macrovesicular steatosis. Recent gene expression studies indicated that tetracycline treatment affects the expression of many genes associated with fatty acid transport and esterification. In this study, we investigated the role of fatty acid transport and esterification in tetracycline-induced steatosis. Intracellular lipid accumulation and the protein expression of fatty acid translocase (FAT or CD36) and diacylglycerol acyltransferase (DGAT) 2 were increased in both mouse liver and HepG2 cells treated with tetracycline at 50 mg/kg (intraperitoneal injection, i.p.) and 100 μM, respectively. Tetracycline increased the cellular uptake of boron-dipyrromethene-labeled C16 fatty acid, which was abolished by CD36 RNA interference. Oleate-induced cellular lipid accumulation was further enhanced by co-incubation with tetracycline. Tetracycline downregulated extracellular signal-regulated kinase (ERK) phosphorylation, which negatively regulated DGAT2 expression. U0126, a specific ERK inhibitor, also increased DGAT2 expression and cellular lipid accumulation. DGAT1 and 2 knock-down with specific small interfering (si)-RNA completely abrogated the steatogenic effect of tetracycline in HepG2 cells. Taken together, our data showed that tetracycline induces lipid accumulation by facilitating fatty acid transport and triglyceride esterification by upregulating CD36 and DGAT2, respectively. PMID:25745068

  14. Salt Contribution to the Flexibility of Single-stranded Nucleic Acid of Finite Length

    Wang, Feng-Hua; Tan, Zhi-Jie

    2012-01-01

    Nucleic acids are negatively charged macromolecules and their structure properties are strongly coupled to metal ions in solutions. In this paper, the salt effects on the flexibility of single stranded (ss) nucleic acid chain ranging from 12 to 120 nucleotides are investigated systematically by the coarse grained Monte Carlo simulations where the salt ions are considered explicitly and the ss chain is modeled with the virtual bond structural model. Our calculations show that, the increase of ion concentration causes the structural collapse of ss chain and multivalent ions are much more efficient in causing such collapse, and trivalent and small divalent ions can both induce more compact state than a random relaxation state. We found that monovalent, divalent and trivalent ions can all overcharge ss chain, and the dominating source for such overcharging changes from ion exclusion volume effect to ion Coulomb correlations. In addition, the predicted Na and Mg dependent persistence length lp of ss nucleic acid a...

  15. Induction of liver metallothionein contribute to the developmental toxicity of valproic acid in rat

    Farid Abolhassani

    2006-03-01

    Full Text Available There is an increased risk of neural tube defects and axial skeletal malformations among infants born by mothers who had received Valproic acid...The aim of the present study is, if administration of valproic acid can induce maternal hepatic Metallothionein (MT synthesis and so secondary decrease of plasma Zn. In the present experimental study, mated rats were divided into four groups of 12 animals each [control, valproic acid (VPA, valproic acid + zinc (VPA+ Zn and Zinc (Zn groups]. The VPA group received 300 mg/kg valproic acid; daily. The control group received an equal volume of 0.9% NaCI. The VPA+ Zn group received 300 mg/kg VPA as well as 30 mg/kg zinc sulfate, and the Zn group received 30 mg/kg zinc sulfate, daily. These drugs were administered intraperitoneally from day 6 through day 15 of gestation. Dams were killed on GD 16 or 20. Blood was drawn to determine plasma zinc; furthermore, maternal liver Zn and MT were also determined. The zinc concentration in the plasma of rats treated with valproic acid was significantly lower than those of the other groups on GD 16 (p=0.004, but liver Zn (p=0.016 and MT (p=0.004 were significantly higher than those of the control group. On GD 20 the incidence of skeletal malformations and neural tube defects tended to be higher in VPA group than VPA+ Zn treated group and no anomalies were seen in the control group. The results from the present experiment support hypothesis that one of biochemical lesions causing the teratogenicity of VPA is a drug induced maternal plasma zinc deficiency secondary to Metallothionein induction in liver.

  16. Apo-10'-lycopenoic acid, a lycopene 1 metabolite, increases sirtuin 1 mRNA and protein levels and decreases hepatic fat accumulation in ob/ob mice

    Lycopene has been shown to be beneficial in protecting against high-fat diet-induced fatty liver. The recent demonstration that lycopene can be converted by carotene 99,10’-oxygenase into a biologically active metabolite, ALA, led us to propose that the function of lycopene can be mediated by ALA. I...

  17. Identification of glyceollin metabolites derived from conjugation with glutathione and glucuronic acid in rats by on-line liquid chromatography-electrospray ionization tandem mass spectrometry

    Glyceollin-related metabolites produced in rats following oral glyceollin administration were screened and identified by precursor and product ion scanning using liquid chromatography, coupled on-line with electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS), to identify all glyceollin me...

  18. LC-MS and GC-MS metabolite profiling of nickel(II) complexes in the latex of the nickel-hyperaccumulating tree Sebertia acuminata and identification of methylated aldaric acid as a new nickel(II) ligand.

    Callahan, Damien L; Roessner, Ute; Dumontet, Vincent; Perrier, Nicolas; Wedd, Anthony G; O'Hair, Richard A J; Baker, Alan J M; Kolev, Spas D

    2008-01-01

    Targeted liquid chromatography-mass spectrometry (LC-MS) technology using size exclusion chromatography and metabolite profiling based on gas chromatography-mass spectrometry (GC-MS) were used to study the nickel-rich latex of the hyperaccumulating tree Sebertia acuminata. More than 120 compounds were detected, 57 of these were subsequently identified. A methylated aldaric acid (2,4,5-trihydroxy-3-methoxy-1,6-hexan-dioic acid) was identified for the first time in biological extracts and its structure was confirmed by 1D and 2D nuclear magnetic resonance (NMR) spectroscopy. After citric acid, it appears to be one of the most abundant small organic molecules present in the latex studied. Nickel(II) complexes of stoichiometry NiII:acid=1:2 were detected for these two acids as well as for malic, itaconic, erythronic, galacturonic, tartaric, aconitic and saccharic acids. These results provide further evidence that organic acids may play an important role in the transport and possibly in the storage of metal ions in hyperaccumulating plants. PMID:17765935

  19. A contribution to the study of mass transfer: uranium extraction from phosphoric acid

    Liquid-liquid extraction of uranium (VI) contained in phosphoric acid is examined. Synergism and thermodynamical equilibria are studied. Influence of drop size, extractant concentration and temperature on reaction kinetics are determined. The experimental study concerns extraction by HDEHP, TOPO and the mixture of both. Reaction mechanisms are interpreted

  20. Rapid and sensitive detection of fipronil and its metabolites in edible oils by solid-phase extraction based on humic acid bonded silica combined with gas chromatography with electron capture detection.

    Peng, Xi-Tian; Li, Yu-Nan; Xia, Hong; Peng, Li-Jun; Feng, Yu-Qi

    2016-06-01

    Solid-phase extraction based on humic acid bonded silica followed by gas chromatography with electron capture detection was developed to determine fipronil and its metabolites in edible oil. To achieve the best extraction performance, we systematically investigated a series of solid-phase extraction parameters. Under the optimized conditions, the method was validated according to linearity, recovery, and precision. Good linearities were obtained with R(2) more than 0.9996 for all analytes. The limits of detection were between 0.3 and 0.5 ng/g, and the recoveries ranged from 83.1 to 104.0% at three spiked concentrations with intra- and interday relative standard deviation values less than 8.7%. Finally, the proposed method was applied to determine fipronil and its metabolites in 11 edible oil samples taken from Wuhan markets. Fipronil was detectable in four samples with concentrations ranging from 3.0 to 5.2 ng/g. In China, the maximum residue limits of fipronil in some vegetables and maize are 20 and 100 ng/g (GB/T 2763-2014), respectively. The residues of fipronil and its metabolites in commercial edible oils might exhibit some potential threat to human health as a result of high consumption of edible oil as part of daily intake. PMID:27280701

  1. Synthesis of 9,12-dioxo-10(Z)-dodecenoic acid, a new fatty acid metabolite derived from 9-hydroperoxy-10,12-octadecadienoic acid in lentil seed (Lens culinaris Medik.)

    Gallasch, B A; Spiteller, G

    2000-09-01

    The previously unknown linoleic acid peroxidation product 9,12-dioxo-10(Z)-decenoic acid (Z5) was detected in lentil seed flour (Lens culinaris Medik.) by electron impact mass spectrometry (EI-MS) after derivatization with pentafluorobenzyl-hydroxylamine-hydrochloride, methylation of acidic groups with diazomethane, and protection of hydroxylic groups with N-methyl-N-trimethylsilyl-trifluoroacetamide. The structure of the natural product was confirmed by synthesis of Z5, 9,12-dioxo-l0(E)-decenoic acid, and derivatives. EI-MS, nuclear magnetic resonance and gas chromatographic data of these compounds and synthetic intermediates are discussed. PMID:11026615

  2. Lactic acid bacteria contribution to gut microbiota complexity: lights and shadows.

    Enrica ePessione

    2012-06-01

    Full Text Available Lactic Acid Bacteria (LAB are ancient organisms that cannot biosynthesize functional cytochromes, and cannot get ATP from respiration. Besides sugar fermentation, they evolved electrogenic decarboxylations and ATP-forming deiminations. The right balance between sugar fermentation and decarboxylation/deimination ensures buffered environments thus enabling LAB to survive in human gastric trait and colonize gut. A complex molecular cross-talk between LAB and host exists. LAB moonlight proteins are made in response to gut stimuli and promote bacterial adhesion to mucosa and stimulate immune cells. Similarly, when LAB are present, human enterocytes activate expression of specific genes only. Furthermore, LAB antagonistic relationships with other microorganisms constitutes the basis for their antiinfective role. Histamine and tyramine are LAB bioactive catabolites that act on the CNS, causing hypertension and allergies. Nevertheless, some LAB biosynthesize both GABA, that has relaxing effect on gut smooth muscles, and beta-phenylethylamine, that controls satiety and mood. Since LAB have reduced amino acid biosynthetic abilities, they developed a sophisticated proteolytic system, that is also involved in antihypertensive and opiod peptide generation from milk proteins.Short-chain fatty acids are glycolytic and phosphoketolase end-products, regulating epithelial cellproliferation and differentiation. Nevertheless, they constitute a supplementary energy source for the host, causing weight gain. Human metabolism can also be affected by anabolic LAB products such as conjugated linoleic acids (CLA. Some CLA isomers reduce cancer cell viability and ameliorate insulin resistance, while others lower the HDL/LDL ratio and modify eicosanoid production, with detrimental health effects.A further appreciated LAB feature is the ability to fix selenium into seleno-cysteine Thus opening interesting perspectives for their utilization as antioxidant nutraceutical

  3. Contributions of tropodithietic acid and biofilm formation to the probiotic activity of Phaeobacter inhibens

    Zhao, Wenjing; Dao, Christine; Karim, Murni; Gomez-Chiarri, Marta; Rowley, David; Nelson, David R.

    2016-01-01

    Background: The probiotic bacterium Phaeobacter inhibens strain S4Sm, isolated from the inner shell surface of a healthy oyster, secretes the antibiotic tropodithietic acid (TDA), is an excellent biofilm former, and increases oyster larvae survival when challenged with bacterial pathogens. In this study, we investigated the specific roles of TDA secretion and biofilm formation in the probiotic activity of S4Sm. Results: Mutations in clpX (ATP-dependent ATPase) and exoP (an exopolysaccharide b...

  4. Contributions of tropodithietic acid and biofilm formation to the probiotic activity of Phaeobacter inhibens

    Zhao, Wenjing; Dao, Christine; Karim, Murni; Gomez-Chiarri, Marta; Rowley, David; Nelson, David R.

    2016-01-01

    Background The probiotic bacterium Phaeobacter inhibens strain S4Sm, isolated from the inner shell surface of a healthy oyster, secretes the antibiotic tropodithietic acid (TDA), is an excellent biofilm former, and increases oyster larvae survival when challenged with bacterial pathogens. In this study, we investigated the specific roles of TDA secretion and biofilm formation in the probiotic activity of S4Sm. Results Mutations in clpX (ATP-dependent ATPase) and exoP (an exopolysaccharide bio...

  5. Chaperone Hsp31 contributes to acid resistance in stationary-phase Escherichia coli.

    Mujacic, Mirna; Baneyx, François

    2007-02-01

    Hsp31, the product of the sigmaS - and sigmaD -dependent hchA gene, is a heat-inducible chaperone implicated in the management of protein misfolding at high temperatures. We show here that Hsp31 plays an important role in the acid resistance of starved Escherichia coli but that it has little influence on oxidative-stress survival. PMID:17158627

  6. Lactic acid bacteria contribution to gut microbiota complexity: lights and shadows

    Pessione, Enrica

    2012-01-01

    Lactic Acid Bacteria (LAB) are ancient organisms that cannot biosynthesize functional cytochromes, and cannot get ATP from respiration. Besides sugar fermentation, they evolved electrogenic decarboxylations and ATP-forming deiminations. The right balance between sugar fermentation and decarboxylation/deimination ensures buffered environments thus enabling LAB to survive in human gastric trait and colonize gut. A complex molecular cross-talk between LAB and host exists. LAB moonlight proteins ...

  7. Pre- and neonatal exposure to lipopolysaccharide or the enteric metabolite, propionic acid, alters development and behavior in adolescent rats in a sexually dimorphic manner.

    Foley, Kelly A; Ossenkopp, Klaus-Peter; Kavaliers, Martin; Macfabe, Derrick F

    2014-01-01

    Alterations in the composition of the gut microbiome and/or immune system function may have a role in the development of autism spectrum disorders (ASD). The current study examined the effects of prenatal and early life administration of lipopolysaccharide (LPS), a bacterial mimetic, and the short chain fatty acid, propionic acid (PPA), a metabolic fermentation product of enteric bacteria, on developmental milestones, locomotor activity, and anxiety-like behavior in adolescent male and female offspring. Pregnant Long-Evans rats were subcutaneously injected once a day with PPA (500 mg/kg) on gestation days G12-16, LPS (50 µg/kg) on G15-16, or vehicle control on G12-16 or G15-16. Male and female offspring were injected with PPA (500 mg/kg) or vehicle twice a day, every second day from postnatal days (P) 10-18. Physical milestones and reflexes were monitored in early life with prenatal PPA and LPS inducing delays in eye opening. Locomotor activity and anxiety were assessed in adolescence (P40-42) in the elevated plus maze (EPM) and open-field. Prenatal and postnatal treatments altered behavior in a sex-specific manner. Prenatal PPA decreased time spent in the centre of the open-field in males and females while prenatal and postnatal PPA increased anxiety behavior on the EPM in female rats. Prenatal LPS did not significantly influence those behaviors. Evidence for the double hit hypothesis was seen as females receiving a double hit of PPA (prenatal and postnatal) displayed increased repetitive behavior in the open-field. These results provide evidence for the hypothesis that by-products of enteric bacteria metabolism such as PPA may contribute to ASD, altering development and behavior in adolescent rats similar to that observed in ASD and other neurodevelopmental disorders. PMID:24466331

  8. Metabolites from intestinal microbes shape Treg

    Geuking, Markus B.; McCoy, Kathy D.; Macpherson, Andrew J

    2013-01-01

    Intestinal bacterial metabolites are an important communication tool between the host immune system and the commensal microbiota to establish mutualism. In a recent paper published in Science, Wendy Garrett and her colleagues report an exciting role of the three most abundant microbial-derived short-chain fatty acids (SCFA), acetic acid, propionic acid and butyric acid, in colonic regulatory T cell (cTreg) homeostasis.

  9. Differential effects of deoxycholic acid versus selenium metabolite methylselenol on cell cycle, apoptosis, and MAP kinase pathway in HCT116 human colon cancer cells

    Introduction: A typical part of the Western diet is a high fat intake that leads to increased levels of fecal bile acids, and these bile acids, primarily deoxycholic acid (DCA) in humans, have been believed to be tumor promoters of colon cancer. The cell growth inhibition induced by bile acid deoxyc...

  10. Contribution of coated humic acids calculated through their surface coverage on nano iron oxides for ofloxacin and norfloxacin sorption

    Sorption of organic contaminants on organo-mineral complexes has been investigated extensively, but the sorption contribution of mineral particles was not properly addressed before calculating KOC, especially for ionic organic contaminants. We measured the surface coverage of a humic acid (HA) on nano iron oxides (n-Fe2O3) in a series of synthesized organo-mineral complexes. The contribution of the coated HA to ofloxacin (OFL) and norfloxacin (NOR) sorption in HA–n-Fe2O3 complexes was over 80% of the total sorption with the surface coverage of 36% and fOC of 1.6%. All the coated HA showed higher sorption to NOR and OFL in comparison to the original HA, suggesting HA fractionation and/or physical re-conformation during organo-mineral complex formation. The decreased KOC with multilayer coating may suggest the importance of site-specific interactions for OFL sorption, while the increased KOC with multilayer coating may suggest the importance of partitioning in hydrophobic region for NOR sorption. - Highlights: • The sorption of nano iron oxides was calculated through their exposed surface area. • The contribution of the coated HA was over 80% of the total sorption. • The decreased KOC with multilayer coating suggests site-specific sorption for OFL. • The increased KOC with multilayer coating suggests NOR partitioning in hydrophobic region. - Coated humic acid showed increased sorption to ionic organic contaminants in comparison to bulk humic acid, and accounted for over 80% of the total sorption at fOC of 1.6%

  11. Quantum chemistry contribution to the understanding of the acid-base phenomena

    The various physical and chemical phenomena involved at the atomic and molecular level of a two material interface, are studied. When intimately in contact, intermolecular range forces are established between their atoms. These forces are divided in two categories: short range and long range forces, and according to the intervention rate, the bonding will be more or less strong. A quantitative approach, in terms of quantum chemistry of the so called acid-base interactions, is proposed and prospects for this approach with the view of enhancing or diminishing the bonding strength between two materials, are discussed

  12. A novel study of screening and confirmation of modafinil, adrafinil and their metabolite modafinilic acid under EI-GC-MS and ESI-LC-MS-MS ionization

    Dubey S; Ahi S; Reddy I; Kaur T; Beotra A; Jain S

    2009-01-01

    Objective: Adrafinil and modafinil have received wide publicity and have become controversial in the sporting world when several athletes were discovered allegedly using these drugs as doping agents. By acknowledging the facts, the World Anti-Doping Agency (WADA) banned these drugs in sports since 2004. The present study explores the possibility of differentiating adrafinil and modafinil and their major metabolites under electron impact ionization in gas chromatograph-mass spectrometer (GC-M...

  13. Exposure of atorvastatin is unchanged but lactone and acid metabolites are increased several-fold in patients with atorvastatin-induced myopathy

    2006-01-01

    Background The most serious side effect from statin treatment is myopathy, which may proceed to rhabdomyolysis. This is the first study to investigate whether the pharmacokinetics of either atorvastatin or its metabolites, or both, is altered in patients with atorvastatin-related myopathy compared with healthy controls. Methods A 24-hour pharmacokinetic investigation was performed in 14 patients with atorvastatin-related myopathy. Relevant polymorphisms in SLCO1B1 (encoding organic ...

  14. Effect of feed restriction on metabolites in cerebrospinal fluid and plasma of dairy cows.

    Laeger, T; Görs, S; Metges, C C; Kuhla, B

    2012-03-01

    Endocrines and metabolites in the circulation act as long-term hunger or satiety signals in the brain during negative energy balance and play an important role in the control of feed intake. These signals also occur in the cerebrospinal fluid (CSF), which surrounds the hypothalamus and brainstem: 2 major centers of feed intake regulation. Thus CSF functions as a transport medium for fuel signals between blood and brain. The CSF metabolite concentrations are mainly under control of the blood-brain barriers, which provide specific carrier molecules facilitating the entry of substances required by the brain and protect the brain from factors that could impair neuronal function. The transport of small molecules such as amino acids (AA) across the blood-brain barriers may be limited by competing AA that share a common transporter for the uptake into brain. Consequently, CSF metabolite concentrations differ from those in blood. Thus it appears likely that central (CSF) rather than peripheral (blood) metabolites act as pivotal signals for the control of feed intake. However, the contribution of putative orexigenic and anorexigenic signals in CSF of cows has not been studied so far. Therefore, the aim of this study was to elucidate associations existing between both plasma and CSF metabolites, each in response to feed restriction-induced negative energy balance. Seven German Holstein dairy cows, between 87 and 96 DIM of the second lactation (milk yield, 27.9 L/d) were fed ad libitum (AL) for 4 d and CSF from the spinal cord and blood from the jugular vein was withdrawn before morning feeding at the fifth day. Subsequently, animals were feed restricted (R) to 50% of the previous AL intake for 4 d and CSF and plasma were collected at the ninth day. Body weight, feed intake, water intake, and milk production were determined. Thirty-one AA, β-hydroxybutyric acid, cholesterol, glucose, lactate, nonesterified fatty acids, urea, and osmolality were measured in both CSF and

  15. Contribution of ants in modifying of soil acidity and particle size distribution

    Morgun, Alexandra; Golichenkov, Maxim

    2015-04-01

    Being a natural body, formed by the influence of biota on the upper layers of the Earth's crust, the soil is the most striking example of biogenic-abiogenic interactions in the biosphere. Invertebrates (especially ants that build soil nests) are important agents that change soil properties in well developed terrestrial ecosystems. Impact of soil microorganisms on soil properties is particularly described in numerous literature and concerns mainly chemical properties and general indicators of soil biological activity. Influence of ants (as representatives of the soil mesofauna) mostly appears as mechanical movement of soil particles and aggregates, and chemical effects caused by concentration of organic matter within the ant's nest. The aim of this research was to evaluate the effect of ants on physical and chemical soil attributes such as particle size distribution and soil acidity. The samples were taken from aerial parts of Lasius niger nests, selected on different elements of the relief (summit position, slope, terrace and floodplain) in the Arkhangelsk region (north of the European part of Russia) and compared with the specimens of the upper horizons of the reference soils. Particle size distribution was determined by laser diffraction method using laser diffraction particle size analyzer «Analysette 22 comfort» (FRITSCH, Germany). The acidity (pH) was determined by potentiometry in water suspension. Particle size distribution of the samples from the nests is more variable as compared to the control samples. For example, the content of 5-10 μm fraction ranges from 9% to 12% in reference soils, while in the anthill samples the variation is from 8% to 15%. Similarly, for 50-250 μm fraction - it ranges from 15% to 18% in reference soils, whereas in anthills - from 6% to 29%. The results of particle size analysis showed that the reference sample on the terrace has silty loam texture and nests soil L. niger are medium loam. The reference soil on the slope is

  16. LTP3 contributes to disease susceptibility in Arabidopsis by enhancing abscisic acid (ABA) biosynthesis.

    Gao, Shan; Guo, Wenya; Feng, Wen; Liu, Liang; Song, Xiaorui; Chen, Jian; Hou, Wei; Zhu, Hongxia; Tang, Saijun; Hu, Jian

    2016-04-01

    Several plant lipid transfer proteins (LTPs) act positively in plant disease resistance. Here, we show that LTP3 (At5g59320), a pathogen and abscisic acid (ABA)-induced gene, negatively regulates plant immunity in Arabidopsis. The overexpression of LTP3 (LTP3-OX) led to an enhanced susceptibility to virulent bacteria and compromised resistance to avirulent bacteria. On infection of LTP3-OX plants with Pseudomonas syringae pv. tomato, genes involved in ABA biosynthesis, NCED3 and AAO3, were highly induced, whereas salicylic acid (SA)-related genes, ICS1 and PR1, were down-regulated. Accordingly, in LTP3-OX plants, we observed increased ABA levels and decreased SA levels relative to the wild-type. We also showed that the LTP3 overexpression-mediated enhanced susceptibility was partially dependent on AAO3. Interestingly, loss of function of LTP3 (ltp3-1) did not affect ABA pathways, but resulted in PR1 gene induction and elevated SA levels, suggesting that LTP3 can negatively regulate SA in an ABA-independent manner. However, a double mutant consisting of ltp3-1 and silent LTP4 (ltp3/ltp4) showed reduced susceptibility to Pseudomonas and down-regulation of ABA biosynthesis genes, suggesting that LTP3 acts in a redundant manner with its closest homologue LTP4 by modulating the ABA pathway. Taken together, our data show that LTP3 is a novel negative regulator of plant immunity which acts through the manipulation of the ABA-SA balance. PMID:26123657

  17. Novel nonadride, heptadride and maleic acid metabolites from the byssochlamic acid producer Byssochlamys fulva IMI 40021 – an insight into the biosynthesis of maleidrides† †Electronic supplementary information (ESI) available: Details of any supplementary information available should be included here. See DOI: 10.1039/c5cc06988b Click here for additional data file.

    Szwalbe, Agnieszka J.; Williams, Katherine; O'Flynn, Daniel E; Andrew M Bailey; Mulholland, Nicholas P.; Vincent, Jason L.; Willis, Christine L.; Cox, Russell J.; Simpson, Thomas J.

    2015-01-01

    The filamentous fungus Byssochlamys fulva strain IMI 40021 produces (+)-byssochlamic acid 1, its novel dihydroanalogue 2 and four related secondary metabolites. Agnestadrides A, 17 and B, 18 constitute a novel class of seven-membered ring, maleic anhydride-containing (hence termed heptadride) natural products. The putative maleic anhydride precursor 5 for both nonadride and heptadride biosynthesis was isolated as a fermentation product for the first time and its structure confirmed by synthes...

  18. Low-molecular-weight metabolite systems chemistry

    Hadacek, Franz; Bachmann, Gert

    2015-01-01

    Low-molecular-weight metabolites (LMWMs) comprise primary or central and a plethora of intermediary or secondary metabolites, all of which are characterized by a molecular weight below 900 Dalton. The latter are especially prominent in sessile higher organisms, such as plants, corals, sponges and fungi, but are produced by all types of microbial organisms too. Common to all of these carbon molecules are oxygen, nitrogen and, to a lesser extent, sulfur, as heteroatoms. The latter can contribut...

  19. β-Orcinol Metabolites from the Lichen Hypotrachyna revoluta

    Panagiota Papadopoulou

    2007-05-01

    Full Text Available Four new β-orcinol metabolites, hypotrachynic acid (1, deoxystictic acid (2, cryptostictinolide (3 and 8 ́-methylconstictic acid (4 along with the metabolites 8 ́-methylstictic acid (5, 8 ́-methylmenegazziaic acid (6, stictic acid (7, 8 ́-ethylstictic acid (8 and atranorin (9, that have been previously described, were isolated for the first time from the tissue extracts of the lichen Hypotrachyna revoluta (Flörke Hale. The structures of the new metabolites were elucidated on the basis of extensive spectroscopic analyses. Radical scavenging activity (RSA of the metabolites isolated in adequate amounts, was evaluated using luminol chemiluminescence and comparison with Trolox®.

  20. Ezrin dephosphorylation/downregulation contributes to ursolic acid-mediated cell death in human leukemia cells

    Ezrin links the actin filaments with the cell membrane and has a functional role in the apoptotic process. It appears clear that ezrin is directly associated with Fas, leading to activation of caspase cascade and cell death. However, the exact role of ezrin in ursolic acid (UA)-induced apoptosis remains unclear. In this study, we show for the first time that UA induces apoptosis in both transformed and primary leukemia cells through dephosphorylation/downregulation of ezrin, association and polarized colocalization of Fas and ezrin, as well as formation of death-inducing signaling complex. These events are dependent on Rho-ROCK1 signaling pathway. Knockdown of ezrin enhanced cell death mediated by UA, whereas overexpression of ezrin attenuated UA-induced apoptosis. Our in vivo study also showed that UA-mediated inhibition of tumor growth of mouse leukemia xenograft model is in association with the dephosphorylation/downregulation of ezrin. Such findings suggest that the cytoskeletal protein ezrin may represent an attractive target for UA-mediated lethality in human leukemia cells

  1. Gene expression and metabolite profiling of developing highbush blueberry fruit indicates transcriptional regulation of flavonoid metabolism and activation of abscisic acid metabolism.

    Zifkin, Michael; Jin, Alena; Ozga, Jocelyn A; Zaharia, L Irina; Schernthaner, Johann P; Gesell, Andreas; Abrams, Suzanne R; Kennedy, James A; Constabel, C Peter

    2012-01-01

    Highbush blueberry (Vaccinium corymbosum) fruits contain substantial quantities of flavonoids, which are implicated in a wide range of health benefits. Although the flavonoid constituents of ripe blueberries are known, the molecular genetics underlying their biosynthesis, localization, and changes that occur during development have not been investigated. Two expressed sequence tag libraries from ripening blueberry fruit were constructed as a resource for gene identification and quantitative real-time reverse transcription-polymerase chain reaction primer design. Gene expression profiling by quantitative real-time reverse transcription-polymerase chain reaction showed that flavonoid biosynthetic transcript abundance followed a tightly regulated biphasic pattern, and transcript profiles were consistent with the abundance of the three major classes of flavonoids. Proanthocyanidins (PAs) and corresponding biosynthetic transcripts encoding anthocyanidin reductase and leucoanthocyanidin reductase were most concentrated in young fruit and localized predominantly to the inner fruit tissue containing the seeds and placentae. Mean PA polymer length was seven to 8.5 subunits, linked predominantly via B-type linkages, and was relatively constant throughout development. Flavonol accumulation and localization patterns were similar to those of the PAs, and the B-ring hydroxylation pattern of both was correlated with flavonoid-3'-hydroxylase transcript abundance. By contrast, anthocyanins accumulated late in maturation, which coincided with a peak in flavonoid-3-O-glycosyltransferase and flavonoid-3'5'-hydroxylase transcripts. Transcripts of VcMYBPA1, which likely encodes an R2R3-MYB transcriptional regulator of PA synthesis, were prominent in both phases of development. Furthermore, the initiation of ripening was accompanied by a substantial rise in abscisic acid, a growth regulator that may be an important component of the ripening process and contribute to the regulation of

  2. Systematic synthesis and anti-inflammatory activity of ω-carboxylated menaquinone derivatives--Investigations on identified and putative vitamin K₂ metabolites.

    Fujii, Shinya; Shimizu, Akitaka; Takeda, Noriaki; Oguchi, Kazuki; Katsurai, Tomoko; Shirakawa, Hitoshi; Komai, Michio; Kagechika, Hiroyuki

    2015-05-15

    Vitamin K is an essential nutrient for blood coagulation and bone homeostasis, and also functions in many physiological processes including inflammation and cancer progression. However, the nature and activities of its metabolites remain unclear. We report here systematic synthesis of ω-carboxylated derivatives of menaquinone (vitamin K2), including previously identified metabolites 5, K acid I (10), and K acid II (12), and evaluation of their inhibitory activity toward LPS-stimulated induction of inflammatory cytokines. These results should contribute to an improved understanding of the biochemistry and pharmacology of vitamin K. PMID:25858455

  3. Improved metabolic health alters host metabolism in parallel with changes in systemic xeno-metabolites of gut origin.

    Caitlin Campbell

    Full Text Available Novel plasma metabolite patterns reflective of improved metabolic health (insulin sensitivity, fitness, reduced body weight were identified before and after a 14-17 wk weight loss and exercise intervention in sedentary, obese insulin-resistant women. To control for potential confounding effects of diet- or microbiome-derived molecules on the systemic metabolome, sampling was during a tightly-controlled feeding test week paradigm. Pairwise and multivariate analysis revealed intervention- and insulin-sensitivity associated: (1 Changes in plasma xeno-metabolites ("non-self" metabolites of dietary or gut microbial origin following an oral glucose tolerance test (e.g. higher post-OGTT propane-1,2,3-tricarboxylate [tricarballylic acid] or in the overnight-fasted state (e.g., lower γ-tocopherol; (2 Increased indices of saturated very long chain fatty acid elongation capacity; (3 Increased post-OGTT α-ketoglutaric acid (α-KG, fasting α-KG inversely correlated with Matsuda index, and altered patterns of malate, pyruvate and glutamine hypothesized to stem from improved mitochondrial efficiency and more robust oxidation of glucose. The results support a working model in which improved metabolic health modifies host metabolism in parallel with altering systemic exposure to xeno-metabolites. This highlights that interpretations regarding the origins of peripheral blood or urinary "signatures" of insulin resistance and metabolic health must consider the potentially important contribution of gut-derived metabolites toward the host's metabolome.

  4. Improved metabolic health alters host metabolism in parallel with changes in systemic xeno-metabolites of gut origin.

    Campbell, Caitlin; Grapov, Dmitry; Fiehn, Oliver; Chandler, Carol J; Burnett, Dustin J; Souza, Elaine C; Casazza, Gretchen A; Gustafson, Mary B; Keim, Nancy L; Newman, John W; Hunter, Gary R; Fernandez, Jose R; Garvey, W Timothy; Harper, Mary-Ellen; Hoppel, Charles L; Meissen, John K; Take, Kohei; Adams, Sean H

    2014-01-01

    Novel plasma metabolite patterns reflective of improved metabolic health (insulin sensitivity, fitness, reduced body weight) were identified before and after a 14-17 wk weight loss and exercise intervention in sedentary, obese insulin-resistant women. To control for potential confounding effects of diet- or microbiome-derived molecules on the systemic metabolome, sampling was during a tightly-controlled feeding test week paradigm. Pairwise and multivariate analysis revealed intervention- and insulin-sensitivity associated: (1) Changes in plasma xeno-metabolites ("non-self" metabolites of dietary or gut microbial origin) following an oral glucose tolerance test (e.g. higher post-OGTT propane-1,2,3-tricarboxylate [tricarballylic acid]) or in the overnight-fasted state (e.g., lower γ-tocopherol); (2) Increased indices of saturated very long chain fatty acid elongation capacity; (3) Increased post-OGTT α-ketoglutaric acid (α-KG), fasting α-KG inversely correlated with Matsuda index, and altered patterns of malate, pyruvate and glutamine hypothesized to stem from improved mitochondrial efficiency and more robust oxidation of glucose. The results support a working model in which improved metabolic health modifies host metabolism in parallel with altering systemic exposure to xeno-metabolites. This highlights that interpretations regarding the origins of peripheral blood or urinary "signatures" of insulin resistance and metabolic health must consider the potentially important contribution of gut-derived metabolites toward the host's metabolome. PMID:24416208

  5. Enteric microbiome metabolites correlate with response to simvastatin treatment.

    Rima Kaddurah-Daouk

    Full Text Available Although statins are widely prescribed medications, there remains considerable variability in therapeutic response. Genetics can explain only part of this variability. Metabolomics is a global biochemical approach that provides powerful tools for mapping pathways implicated in disease and in response to treatment. Metabolomics captures net interactions between genome, microbiome and the environment. In this study, we used a targeted GC-MS metabolomics platform to measure a panel of metabolites within cholesterol synthesis, dietary sterol absorption, and bile acid formation to determine metabolite signatures that may predict variation in statin LDL-C lowering efficacy. Measurements were performed in two subsets of the total study population in the Cholesterol and Pharmacogenetics (CAP study: Full Range of Response (FR, and Good and Poor Responders (GPR were 100 individuals randomly selected from across the entire range of LDL-C responses in CAP. GPR were 48 individuals, 24 each from the top and bottom 10% of the LDL-C response distribution matched for body mass index, race, and gender. We identified three secondary, bacterial-derived bile acids that contribute to predicting the magnitude of statin-induced LDL-C lowering in good responders. Bile acids and statins share transporters in the liver and intestine; we observed that increased plasma concentration of simvastatin positively correlates with higher levels of several secondary bile acids. Genetic analysis of these subjects identified associations between levels of seven bile acids and a single nucleotide polymorphism (SNP, rs4149056, in the gene encoding the organic anion transporter SLCO1B1. These findings, along with recently published results that the gut microbiome plays an important role in cardiovascular disease, indicate that interactions between genome, gut microbiome and environmental influences should be considered in the study and management of cardiovascular disease. Metabolic

  6. Determination of glyphosate residuals and of their metabolite Aminomethyl-Phosphonic acid in waters, by means of liquid chromatography of high efficiency with post-column derivation and fluorescence detection

    The glyphosate is a no selective herbicide largely used in the world in order to control annual and perennial weeds. Its principal metabolite in soils and waters is the Aminomethyl-Phosphonic Acid (AMPA) formed by micro organism's action. This herbicide is used in Colombia in high doses to illegal crops eradication of coca and Amapola and like natural accelerator in sugar cane, constituting an environmental and social problem for the country, being necessary the evaluation of glyphosate residues in different matrices. This study describes the validation of the analytical methodology for the simultaneous determination of glyphosate and its metabolite AMPA in waters of some Colombian regions. The experimental procedure pointed out two main steps: the first one was a cleaning, extraction and concentration step by solid phase extraction; the second step is the separation, identification and quantification of the compounds by High Performance Liquid Chromatography (HPLC) with post-column derivation and fluorescence detection. The results of the validation show that the methodology is specific, selective, precise and robust with linear calibration curve in the linear range between 10 and 750 μg/L, with limits of detection of 0.8 μg/L and limits of quantification of 2 μg/L for the two analyses. The recoveries are in the order of 73% for glyphosate and 70% for AMPA. More over analysis results are presented for water samples of some country regions where glyphosate is applied in different doses with different purposes, finding residues of the herbicide and its metabolite in concentrations above the allowed values in drinking waters for pesticides of toxicology category IV, like the glyphosate, according with the Colombian legislation

  7. 白酒生产中乳酸菌的分布及主要代谢产物%The distribution of lactic acid bacteria in traditional liquor production and main metabolites

    孙超; 刘勇

    2012-01-01

    乳酸菌是发酵产乳酸的一类革兰氏阳性菌,是大曲中的主要微生物菌系,对大曲中酯的形成是有利的,但在有些香型白酒(如清香、浓香型白酒)酿造中要控制或降低乳酸菌的作用.该文介绍了白酒生产中乳酸菌的分布,乳酸菌所产主要代谢产物的代谢途径及其对白酒的影响.%Lactic acid bacterium is a kind of gram-positive bacterium that generates lactic fermentation,which is the main microbial strains in daqu and in the form of daqu ester is favorable.But in some liquor flavor types (such as faint scent, luzhcu-flavor liquor) production we need to control or reduce the role of lactic acid bacteria. This paper introduces the distribution of lactic acid bacteria in liquor production, metabolic pathway of main metabolites of lactic acid bacteria and its influence for liquor.

  8. Dietary omega-3 fatty acids modulate the eicosanoid profile in man primarily via the CYP-epoxygenase pathway[S

    Fischer, Robert; Konkel, Anne; Mehling, Heidrun; Blossey, Katrin; Gapelyuk, Andrej; Wessel, Niels; von Schacky, Clemens; Dechend, Ralf; Muller, Dominik N.; Rothe, Michael; Luft, Friedrich C.; Weylandt, Karsten; Schunck, Wolf-Hagen

    2014-01-01

    Cytochrome P450 (CYP)-dependent metabolites of arachidonic acid (AA) contribute to the regulation of cardiovascular function. CYP enzymes also accept EPA and DHA to yield more potent vasodilatory and potentially anti-arrhythmic metabolites, suggesting that the endogenous CYP-eicosanoid profile can be favorably shifted by dietary omega-3 fatty acids. To test this hypothesis, 20 healthy volunteers were treated with an EPA/DHA supplement and analyzed for concomitant changes in the circulatory an...

  9. In vitro release of arachidonic acid metabolites, glutathione peroxidase, and oxygen-free radicals from platelets of asthmatic patients with and without aspirin intolerance.

    Plaza, V.; J. Prat; Rosellò, J.; Ballester, E; Ramis, I; Mullol, J; Gelpí, E; Vives-Corrons, J. L.; Picado, C.

    1995-01-01

    BACKGROUND--An abnormal platelet release of oxygen-free radicals has been described in acetylsalicylic acid (aspirin)-induced asthma, a finding which might suggest the existence of an intrinsic, specific platelet abnormality of arachidonic acid metabolism in these patients. The objective of this study was to evaluate platelet arachidonic acid metabolism in asthmatic patients with or without intolerance to aspirin. METHODS--Thirty subjects distributed into three groups were studied: group 1, 1...

  10. Validation of UHPLC-MS/MS methods for the determination of kaempferol and its metabolite 4-hydroxyphenyl acetic acid, and application to in vitro blood-brain barrier and intestinal drug permeability studies.

    Moradi-Afrapoli, Fahimeh; Oufir, Mouhssin; Walter, Fruzsina R; Deli, Maria A; Smiesko, Martin; Zabela, Volha; Butterweck, Veronika; Hamburger, Matthias

    2016-09-01

    Sedative and anxiolytic-like properties of flavonoids such as kaempferol and quercetin, and of some of their intestinal metabolites, have been demonstrated in pharmacological studies. However, routes of administration were shown to be critical for observing in vivo activity. Therefore, the ability to cross intestinal and blood-brain barriers was assessed in cell-based models for kaempferol (KMF), and for the major intestinal metabolite of KMF, 4-hydroxyphenylacetic acid (4-HPAA). Intestinal transport studies were performed with Caco-2 cells, and blood-brain barrier transport studies with an immortalized monoculture human model and a primary triple-co-culture rat model. UHPLC-MS/MS methods for KMF and 4-HPAA in Ringer-HEPES buffer and in Hank's balanced salt solution were validated according to industry guidelines. For all methods, calibration curves were fitted by least-squares quadratic regression with 1/X(2) as weighing factor, and mean coefficients of determination (R(2)) were >0.99. Data obtained with all barrier models showed high intestinal and blood-brain barrier permeation of KMF, and no permeability of 4-HPAA, when compared to barrier integrity markers. PMID:27281582

  11. Analysis of variability of concentrations of valproic acid (VPA and its selected metabolites in the blood serum of patients treated with VPA and patients hospitalized because of VPA poisoning

    Jolanta Wilimowska

    2015-03-01

    Full Text Available Aim of the study: To compare the metabolic profile of valproic acid (VPA in the studied groups of cases through an analysis of variability of concentrations of VPA with its selected metabolites (2-ene-VPA, 4-ene-VPA, 3-keto-VPA. Study material : Blood serum samples collected from 27 patients treated with VPA drugs in the Psychiatry Unit and in the Neurology and Cerebral Strokes Unit at the Ludwik Rydygier Provincial Specialist Hospital in Krakow, and blood serum samples collected from 26 patients hospitalized because of suspected acute VPA poisoning at the Toxicology Department, Chair of Toxicology and Environmental Diseases, Jagiellonian University Medical College in Krakow. Results and conclusions : The analysis of concentrations of VPA and its selected metabolites has shown that the metabolic profile of VPA determined in cases of acute poisoning is different from cases of VPA therapy. One of VPA’s metabolic pathways – the process of desaturation – is unchanged in acute poisoning and prevails over the process of -oxidation. The ingestion of toxic VPA doses results in an increased formation of 4-ene-VPA, proportional to an increase in VPA concentration. Acute VPA poisoning involves the saturation of VPA’s metabolic transformations at the stage of -oxidation. The process of oxidation of 2-ene-VPA to 3-keto-VPA is slowed down after the ingestion of toxic doses.

  12. Basal efflux of bile acids contributes to drug-induced bile acid-dependent hepatocyte toxicity in rat sandwich-cultured hepatocytes.

    Susukida, Takeshi; Sekine, Shuichi; Ogimura, Eiichiro; Aoki, Shigeki; Oizumi, Kumiko; Horie, Toshiharu; Ito, Kousei

    2015-10-01

    The bile salt export pump (BSEP or Bsep) functions as an apical transporter to eliminate bile acids (BAs) from hepatocytes into the bile. BSEP or Bsep inhibitors engender BA retention, suggested as an underlying mechanism of cholestatic drug-induced liver injury. We previously reported a method to evaluate BSEP-mediated BA-dependent hepatocyte toxicity by using sandwich-cultured hepatocytes (SCHs). However, basal efflux transporters, including multidrug resistance-associated proteins (MRP or Mrp) 3 and 4, also participate in BA efflux. This study examined the contribution of basal efflux transporters to BA-dependent hepatocyte toxicity in rat SCHs. The apical efflux of [(3)H]taurocholic acid (TC) was potently inhibited by 10 μM cyclosporine A (CsA), with later inhibition of basal [(3)H]TC efflux, while MK571 simultaneously inhibited both apical and basal [(3)H]TC efflux. CsA-induced BA-dependent hepatocyte toxicity was 30% at most at 10 μM CsA and ∼60% at 50 μM, while MK571 exacerbated hepatocyte toxicity at concentrations of ≥50 μM. Quinidine inhibited only basal [(3)H]TC efflux and showed BA-dependent hepatocyte toxicity in rat SCHs. Hence, inhibition of basal efflux transporters as well as Bsep may precipitate BA-dependent hepatocyte toxicity in rat SCHs. PMID:26055650

  13. Deoxycholic Acid and Selenium Metabolite Methylselenol Exert Common and Distinct Effects on Cell Cycle, Apoptosis, and MAP Kinase Pathway in HCT116 Human Colon Cancer Cells

    Bile acid deoxycholic acid (DCA) is a known tumor promoter in colon tumor development. The cell growth inhibition induced by DCA may cause compensatory hyperproliferation of colonic epithelial cells and provide selection for subpopulations of cells resistant to DCA’s inhibitory effect. These survivi...

  14. Potato Patatin Generates Short-Chain Fatty Acids from Milk Fat that Contribute to Flavour Development in Cheese Ripening.

    Spelbrink, Robin E J; Lensing, Hellen; Egmond, Maarten R; Giuseppin, Marco L F

    2015-05-01

    The potato lipase, patatin, has long been thought of as essentially inactive towards triacylglycerols. Recently, technology has been developed to isolate potato proteins in native form as food ingredients at industrial scale. Characterisation of native patatin obtained in this way revealed that this enzyme activity towards triacylglycerols has been underestimated. This enables the application of patatin in cheese ripening, which is described in this study. When patatin is added to milk during cheese making, the lipase preferentially releases short-chain fatty acids that contribute to cheese flavour in a dose-dependent manner. Fortuitously, the lipase activity is found mainly in the curd. The release of the short-chain fatty acids matches the activity profile of patatin towards homotriacylglycerols of defined chain length. Residual patatin in the whey fraction can be inactivated effectively by heat treatment that follows Arrhenius kinetics. The results are discussed in terms of cheese making, patatin substrate preference and implications for the use of patatin more generally in food emulsions. PMID:25809992

  15. Strontium isotope quantification of siderite, brine and acid mine drainage contributions to abandoned gas well discharges in the Appalachian Plateau

    Chapman, Elizabeth C. [Univ. of Pittsburgh, PA (United States). Dept. of Geology and Planetary Science; Capo, Rosemary C. [Univ. of Pittsburgh, PA (United States). Dept. of Geology and Planetary Science; Stewart, Brian W. [Univ. of Pittsburgh, PA (United States). Dept. of Geology and Planetary Science; Hedin, Robert S. [Hedin Environmental, Pittsburgh, PA (United States); Weaver, Theodore J. [Hedin Environmental, Pittsburgh, PA (United States); Edenborn, Harry M. [National Energy Technology Lab. (NETL), Pittsburgh, PA, (United States)

    2013-04-01

    Unplugged abandoned oil and gas wells in the Appalachian region can serve as conduits for the movement of waters impacted by fossil fuel extraction. Strontium isotope and geochemical analysis indicate that artesian discharges of water with high total dissolved solids (TDS) from a series of gas wells in western Pennsylvania result from the infiltration of acidic, low Fe (Fe < 10 mg/L) coal mine drainage (AMD) into shallow, siderite (iron carbonate)-cemented sandstone aquifers. The acidity from the AMD promotes dissolution of the carbonate, and metal- and sulfate-contaminated waters rise to the surface through compromised abandoned gas well casings. Strontium isotope mixing models suggest that neither upward migration of oil and gas brines from Devonian reservoirs associated with the wells nor dissolution of abundant nodular siderite present in the mine spoil through which recharge water percolates contribute significantly to the artesian gas well discharges. Natural Sr isotope composition can be a sensitive tool in the characterization of complex groundwater interactions and can be used to distinguish between inputs from deep and shallow contamination sources, as well as between groundwater and mineralogically similar but stratigraphically distinct rock units. This is of particular relevance to regions such as the Appalachian Basin, where a legacy of coal, oil and gas exploration is coupled with ongoing and future natural gas drilling into deep reservoirs.

  16. Human urinary metabolites of 3-(2',4',5'-triethoxybenzoyl)propionic acid, a new biliary smooth muscle relaxant with choleretic activity.

    Kobayashi, T; Kanai, Y; Tanayama, S

    1978-12-01

    Biotransformation of 3-(2',4',5'-triethoxybenzoyl)propionic acid (tri-, ethoxybenzoylpropionate) was studied in man using isotopic cluster technique. From the 24 h urine of a male volunteer given an equimolar mixture of non-labelled and pentadeuterium-labelled triethoxybenzoylpropionate (1 mg/kg) orally, the parent drug, 3-(2',5'-diethoxy-4'-hydroxybenzoyl)propionic acid (4'-phenol) and 3-(2',4'-diethoxy-5'-hydroxybenzoyl)propionic acid (5'-phenol) were isolated and identified by g.l.c.-mass spectrometry. 2. Urinary components were also quantified by mass chromatography: the parent drug (67.4% of dose) and 4'- and 5'-phenols in a mixture (20.3%). More than 80% of urinary triethoxybenzoylpropionate was present as its glucuronic acid ester, as evidenced by using beta-glucuronidase and saccharo-1,4-lactone, a specific inhibitor of the enzyme. Almost all of 4'- and 5'-phenols appeared as conjugates with glucuronic acid and/or sulphuric acid. These findings indicated that in man triethoxybenzoylpropionate was largely conjugated with glucuronic acid and in part underwent deethylation followed by conjugation for elimination mainly in urine. PMID:726517

  17. Metabolite profiling of Arabidopsis seedlings in response to exogenous sinalbin and sulfur deficiency.

    Zhang, Jixiu; Sun, Xiumei; Zhang, Zhiping; Ni, Yuwen; Zhang, Qing; Liang, Xinmiao; Xiao, Hongbin; Chen, Jiping; Tokuhisa, James G

    2011-10-01

    In order to determine how plant uptake of a sulfur-rich secondary metabolite, sinalbin, affects the metabolic profile of sulfur-deficient plants, gas chromatography time-of-flight mass spectrometry (GC-TOF-MS), in combination with liquid chromatography-mass spectrometry (LC-MS), was used to survey the metabolome of Arabidopsis seedlings grown in nutrient media under different sulfur conditions. The growth media had either sufficient inorganic sulfur for normal plant growth or insufficient inorganic sulfur in the presence or absence of supplementation with organic sulfur in the form of sinalbin (p-hydroxybenzylglucosinolate). A total of 90 metabolites were identified by GC-TOF-MS and their levels were compared across the three treatments. Of the identified compounds, 21 showed similar responses in plants that were either sulfur deficient or sinalbin supplemented compared to sulfur-sufficient plants, while 12 metabolites differed in abundance only in sulfur-deficient plants. Twelve metabolites accumulated to higher levels in sinalbin-supplemented than in the sulfur-sufficient plants. Secondary metabolites such as flavonol conjugates, sinapinic acid esters and glucosinolates, were identified by LC-MS and their corresponding mass fragmentation patterns were determined. Under sinalbin-supplemented conditions, sinalbin was taken up by Arabidopsis and contributed to the endogenous formation of glucosinolates. Additionally, levels of flavonol glycosides and sinapinic acid esters increased while levels of flavonol diglycosides with glucose attached to the 3-position were reduced. The exogenously administered sinalbin resulted in inhibition of root and hypocotyl growth and markedly influenced metabolite profiles, compared to control and sulfur-deficient plants. These results indicate that, under sulfur deficient conditions, glucosinolates can be a sulfur source for plants. This investigation defines an opportunity to elucidate the mechanism of glucosinolate degradation in

  18. Studies on the secondary metabolites from the Indian gorgonian Subergorgia suberosa: Isolation and characterization of four analogues of the cardiotoxin subergorgic acid

    Parameswaran, P.S.; Naik, C.G.; Kamat, S.Y.; Puar, M.S.; Das, Pradip; Hegde, V.R.

    Chemical investigation of the methanol extract of the Indian Ocean gorgonian coral Subergorgia suberosa resulted in isolation and identification of four novel compounds 2-5. Structural investigation revealed compound 1 to be subergorgic acid...

  19. Biodegradation of Green HE4B: Co-substrate effect, biotransformation enzymes and metabolite toxicity analysis.

    Kalme, S D; Jadhav, S U; Parshetti, G K; Govindwar, S P

    2010-06-01

    A high exhaust reactive dye, Green HE4B (GHE4B) was 98% degraded in nutrient medium by Pseudomonas desmolyticum NCIM 2112 (pd2112) within 72 h at static condition. Decolorization time in synthetic 10 g/l molasses. Addition of 5 g/l peptone to NaCl medium had reduced decolorization time from 108 to 72 h. Beef extract do not contribute more to the inducing effect of peptone, however it is a good co-substrate in sucrose or urea containing NaCl medium. Intracellular lignin peroxidase (Lip), laccase and tyrosinase activities were induced by 150, 355 and 212%, respectively till maximum dye removal took place. Aminopyrine N-demethylase (AND) and dichlorophenol indophenol reductase (DCIP-reductase) activities in pd2112 were induced by 130 and 20%, respectively at 72 h of incubation during GHE4B decolorization. By high performance liquid chromatography (HPLC) analysis, 4-hydroxybenzene sulfonic acid and 4-amino, 6-hydroxynaphthalene 2-sulfonic acids were identified as metabolites formed during 24-72 h incubation. Fourier transform infrared spectroscopy (FTIR) analysis supports the formation of these aromatic amines. pd2112, aerobically degraded GHE4B metabolites (formed at static condition) showing stationary phase of 6 days. There was no germination inhibition of Sorghum bicolor and Triticum aestivum by GHE4B metabolites at 3,000 ppm concentration however untreated dye showed germination inhibition at the same concentration. GHE4B metabolites did not show any microbial toxicity at 10,000 ppm concentration. PMID:23100822

  20. Identification of two main urinary metabolites of [14C]omeprazole in humans

    The excretion and metabolism of [14C]omeprazole given orally as a suspension was studied in 10 healthy male subjects. An average of 79% of the dose was recovered in the urine in 96 hr, with most of the radioactivity (76% of dose) being eliminated in the first 24 hr. Pooled urine (0-2 hr) from five subjects, containing about 47% of the dose, was analyzed by reverse phase gradient elution LC with radioisotope detection. Omeprazole was completely metabolized to at least six metabolites. The two major metabolites were extensively purified by LC and their structures were determined by MS with derivatization and use of stable isotopes, 1H NMR, and comparison with synthetic references. They were formed by hydroxylation of a methyl group in the pyridine ring, followed by further oxidation of the alcohol to the corresponding carboxylic acid. Both metabolites retained the sulfoxide group of omeprazole, rendering them as unstable as the parent compound at pH less than 7. They accounted for approximately 28% (hydroxyomeprazole) and 23% (omeprazole acid) of the amount excreted in the 0-2-hr collection interval. Based on in vitro studies with the synthetic metabolites in isolated gastric glands, it is unlikely that M1 and M2 will contribute to the pharmacological effect of omeprazole in humans

  1. Enteric Bacterial Metabolites Propionic and Butyric Acid Modulate Gene Expression, Including CREB-Dependent Catecholaminergic Neurotransmission, in PC12 Cells - Possible Relevance to Autism Spectrum Disorders

    Nankova, Bistra B; Agarwal, Raj; MacFabe, Derrick F.; La Gamma, Edmund F.

    2014-01-01

    Alterations in gut microbiome composition have an emerging role in health and disease including brain function and behavior. Short chain fatty acids (SCFA) like propionic (PPA), and butyric acid (BA), which are present in diet and are fermentation products of many gastrointestinal bacteria, are showing increasing importance in host health, but also may be environmental contributors in neurodevelopmental disorders including autism spectrum disorders (ASD). Further to this we have shown SCFA ad...

  2. Inhibition of endocannabinoid metabolism by the metabolites of ibuprofen and flurbiprofen.

    Jessica Karlsson

    Full Text Available In addition to their effects upon prostaglandin synthesis, the non-steroidal anti-inflammatory drugs ibuprofen and flurbiprofen inhibit the metabolism of the endocannabinoids 2-arachidonoylglycerol (2-AG and anandamide (AEA by cyclooxygenase-2 (COX-2 and fatty acid amide hydrolase (FAAH, respectively. Here, we investigated whether these effects upon endocannabinoid metabolism are shared by the main metabolites of ibuprofen and flurbiprofen.COX activities were measured via changes in oxygen consumption due to oxygenation of arachidonic acid (for COX-1 and arachidonic acid and 2-AG (for COX-2. FAAH activity was quantified by measuring hydrolysis of tritium labelled AEA in rat brain homogenates. The ability of ibuprofen and flurbiprofen to inhibit COX-2-catalysed oxygenation of 2-AG at lower concentrations than the oxygenation of arachidonic acid was seen with 4'-hydroxyflurbiprofen and possibly also 3'-hydroxyibuprofen, albeit at lower potencies than the parent compounds. All ibuprofen and flurbiprofen metabolites retained the ability to inhibit FAAH in a pH-dependent manner, although the potency was lower than seen with the parent compounds.It is concluded that the primary metabolites of ibuprofen and flurbiprofen retain some of the properties of the parent compound with respect to inhibition of endocannabinoid metabolism. However, these effects are unlikely to contribute to the actions of the parent compounds in vivo.

  3. Epigenome targeting by probiotic metabolites

    Licciardi Paul V

    2010-12-01

    Full Text Available Abstract Background The intestinal microbiota plays an important role in immune development and homeostasis. A disturbed microbiota during early infancy is associated with an increased risk of developing inflammatory and allergic diseases later in life. The mechanisms underlying these effects are poorly understood but are likely to involve alterations in microbial production of fermentation-derived metabolites, which have potent immune modulating properties and are required for maintenance of healthy mucosal immune responses. Probiotics are beneficial bacteria that have the capacity to alter the composition of bacterial species in the intestine that can in turn influence the production of fermentation-derived metabolites. Principal among these metabolites are the short-chain fatty acids butyrate and acetate that have potent anti-inflammatory activities important in regulating immune function at the intestinal mucosal surface. Therefore strategies aimed at restoring the microbiota profile may be effective in the prevention or treatment of allergic and inflammatory diseases. Presentation of the hypothesis Probiotic bacteria have diverse effects including altering microbiota composition, regulating epithelial cell barrier function and modulating of immune responses. The precise molecular mechanisms mediating these probiotic effects are not well understood. Short-chain fatty acids such as butyrate are a class of histone deacetylase inhibitors important in the epigenetic control of host cell responses. It is hypothesized that the biological function of probiotics may be a result of epigenetic modifications that may explain the wide range of effects observed. Studies delineating the effects of probiotics on short-chain fatty acid production and the epigenetic actions of short-chain fatty acids will assist in understanding the association between microbiota and allergic or autoimmune disorders. Testing the hypothesis We propose that treatment with

  4. Secondary metabolites in grasses: characterization and biological activity

    Aldo Tava

    2007-01-01

    In a series of studies dealing on the nutritional value of forage species, more attention was focussed on several compounds, named secondary metabolites, that are important in determining nutritional characteristics. Secondary metabolites are compounds detected in the green materials in low concentration compared to primary metabolites (proteins, sugars, lipids, fibers), but of fundamental importance for the plant physiology. The possess several biological activities and this contribute to th...

  5. Synthesis of an Albendazole Metabolite: Characterization and HPLC Determination

    Mahler, Graciela; Davyt, Danilo; Gordon, Sandra; Incerti, Marcelo; Nunez, Ivana; Pezaroglo, Horacio; Scarone, Laura; Serra, Gloria; Silvera, Mauricio; Manta, Eduardo

    2008-01-01

    In this laboratory activity, students are introduced to the synthesis of an albendazole metabolite obtained by a sulfide oxidation reaction. Albendazole as well as its metabolite, albendazole sulfoxide, are used as anthelmintic drugs. The oxidation reagent is H[subscript 2]O[subscript 2] in acetic acid. The reaction is environmental friendly,…

  6. Effect of acetic acid feeding on the circadian changes in glycogen and metabolites of glucose and lipid in liver and skeletal muscle of rats.

    Fushimi, Takashi; Sato, Yuzo

    2005-11-01

    The aim of the present study is to investigate the effect of acetic acid feeding on the circadian changes in glycogen concentration in liver and skeletal muscle. Rats were provided meal once daily (09.00-13.00 hours) for 10 d. On the 11th day, they were either killed immediately or given 9 g diet containing either 0 (control) or 0.7 g/kg-diet acetic acid beginning at 09.00 hours for 4 h, as in the previous regimen. Rats in the fed group were killed at 4, 8 or 24 h after the start of feeding. At 4 h after the start of feeding, the acetic acid group had significantly greater liver and gastrocnemius muscle glycogen concentrations (Psupercompensation, a large increase in the glycogen level that is beneficial in improving performance, in liver and skeletal muscle by transitory inhibition of glycolysis. Further, we indicate the possibility of a transient enhancement of fatty acid oxidation in liver by acetic acid feeding. PMID:16277773

  7. Microbial contributions to coupled arsenic and sulfur cycling in the acid-sulfide hot spring Champagne Pool, New Zealand

    Katrin eHug

    2014-11-01

    Full Text Available Acid-sulfide hot springs are analogs of early Earth geothermal systems where microbial metal(loid resistance likely first evolved. Arsenic is a metalloid enriched in the acid-sulfide hot spring Champagne Pool (Waiotapu, New Zealand. Arsenic speciation in Champagne Pool follows reaction paths not yet fully understood with respect to biotic contributions and coupling to biogeochemical sulfur cycling. Here we present quantitative arsenic speciation from Champagne Pool, finding arsenite dominant in the pool, rim and outflow channel (55-75% total arsenic, and dithio- and trithioarsenates ubiquitously present as 18-25% total arsenic. In the outflow channel, dimethylmonothioarsenate comprised ≤9% total arsenic, while on the outflow terrace thioarsenates were present at 55% total arsenic. We also quantified sulfide, thiosulfate, sulfate and elemental sulfur, finding sulfide and sulfate as major species in the pool and outflow terrace, respectively. Elemental sulfur reached a maximum at the terrace. Phylogenetic analysis of 16S rRNA genes from metagenomic sequencing revealed the dominance of Sulfurihydrogenibium at all sites and an increased archaeal population at the rim and outflow channel. Several phylotypes were found closely related to known sulfur- and sulfide-oxidizers, as well as sulfur- and sulfate-reducers. Bioinformatic analysis revealed genes underpinning sulfur redox transformations, consistent with sulfur speciation data, and illustrating a microbial role in sulfur-dependent transformation of arsenite to thioarsenate. Metagenomic analysis also revealed genes encoding for arsenate reductase at all sites, reflecting the ubiquity of thioarsenate and a need for microbial arsenate resistance despite anoxic conditions. Absence of the arsenite oxidase gene, aio, at all sites suggests prioritization of arsenite detoxification over coupling to energy conservation. Finally, detection of methyl arsenic in the outflow channel, in conjunction with

  8. Electrogenerated chemiluminescence quenching of Ru(bpy)32+ (bpy=2,2′-bipyridine) in the presence of acetaminophen, salicylic acid and their metabolites

    Quenching of Ru(bpy) 32+ (bpy=2,2′-bipyridine) coreactant electrogenerated chemiluminescence (ECL) has been observed in the presence of acetaminophen, salicylic acid and related complexes. However, no quenching is observed with the acetylsalicylic acid. In most instances, quenching is observed with 100-fold excess of quencher (compared to ECL luminophore) with complete quenching observed between 10,000 and 100,000 fold excess. Fluorescence and UV–vis experiments coupled with bulk electrolysis support the formation of benzoquinone products upon electrochemical oxidation. The mechanism of quenching may involve the interaction of the electrochemically generated benzoquinone species with (i) the ⁎Ru(bpy)32+ excited state or (ii) highly energetic coreactant radicals. - Highlights: ▶ Efficient quenching of the electrogenerated chemiluminescence is observed. ▶ Acetaminophen, salicylic acid and related compounds can be detected. ▶ The mechanism of quenching involves benzoquinones formed upon electrolysis.

  9. Oxygenated metabolites of anandamide and 2-arachidonoylglycerol : conformational analysis and interaction with cannabinoid receptors, membrane transporter, and fatty acid amide hydrolase

    Vliegenthart, J.F.G.; Stelt, M. van der; Kuik, J.A. van; Zadelhoff, G. van; Leeflang, B.R.; Veldink, G.A.; Finazzi Agrò, A.; Maccarrone, M.

    2002-01-01

    This study was aimed at finding structural requirements for the interaction of the acyl chain of endocannabinoids with cannabinoid receptors, membrane transporter protein, and fatty acid amide hydrolase (FAAH). To this end, the flexibility of the acyl chain was restricted by introduction of an 1-hyd

  10. Effects Of Haloacetic Acids and their major metabolites in a Mouse Embryonic Stem Cell Adherent Cell Differentiation and Cytotoxicity (ACDC) Assay

    The haloacetic acids (HAAs) are a class of chemicals produced by disinfection of drinking water. Many of the HAAs are developmental toxicants when administered to rodents producing a variety of developmental effects. We have previously shown that the HAAs can produce direct effec...

  11. Natural product inhibitors of fatty acid biosynthesis: synthesis of the marine microbial metabolites pseudopyronines A and B and evaluation of their anti-infective activities

    Giddens, Anna C.; Nielsen, Lone; Boshoff, Helena I.;

    2007-01-01

    of pathogenic microorganisms and were found to exhibit good potency (IC50≥0.46 μg/mL) and selectivity towards Leishmania donovani. Several of the compounds inhibited recombinant fatty acid biosynthesis enzymes from both Plasmodium falciparum and Mycobacterium tuberculosis, validating these targets in the search...

  12. Identification of a classical mutant in the industrial host Aspergillus niger by systems genetics: LaeA is required for citric acid production and regulates the formation of some secondary metabolites

    Liu, Jing; Arentshorst, Mark; Nair, Deepa; Dai, Ziyu; Baker, Scott E.; Frisvad, Jens; Nielsen, Kristian F.; Punt, Peter J.; Ram, Arthur F.

    2016-01-11

    Rapid acidification of the culture medium by the production of organic acids and the production of acid-induced proteases are key characteristics of the filamentous fungus Aspergillus niger. The D15 mutant of A. niger is non-acidifying mutant and used often for the expression of recombinant proteins in A. niger, because of its reduced production of extracellular proteases under non-acidic conditions. In this study, the D15 mutant is characterized in detail. Strongly reduced levels of citric and oxalic acid were observed in the D15 mutant both in shake flask cultures and in controlled batch cultivations. To identify the mutation in the D15 mutant, we successfully combined high-throughput sequencing (Illumina) with bulk segregant analysis. Because of the lack of a sexual cycle for A. niger, the parasexual cycle was used to generate a pool of segregants. From the 52 single nucleotide polymorphisms (SNPs) between the parental strains, three SNPs were homozygous in the genomic DNA of pool of segregants. These three SNPs mapped to all the right arm of chromosome II, indicating that this region contains the genetic locus affecting the phenotype related to acid production. Of the three SNPs, one mutation resulted in a missense mutation in the gene encoding the A. niger homologue of the A. nidulans methyltransferase gene laeA. Complementation analysis of the original mutant with the laeA gene and targeted disruption of laeA further confirmed that LaeA is involved in citric acid production in A. niger lab (N402) and citric acid production strains (ATCC 11414). Analysis of the secondary metabolite (SM) profile of the laeA mutants indicate that LaeA is required for the production of several SMs (asperrubrol, atromentin and JBIR86), but deletion of laeA also resulted in the presence of SMs (aspernigrin A/B and BMS-192548) that were not detected in the wild-type strain. The levels of ten other SMs were not strongly affected as a result of laeA deletion indicating that only a

  13. OMEGA-3 fatty acids contribute to plaque stability differentially affecting the release of matrix metalloproteinases and tissue inhibitors of metalloproteinases by human monocytes/macrophages in culture

    Massaro, Marika; Scoditti, Egeria; Carluccio, Maria Annunziata; Storelli, Carlo; Distante, Alessandro; Martines, Giuseppe

    2008-01-01

    Objectives. High intakes of omega-3 fatty acids has been associated with protection from plaque rupture. The secretion of metalloproteinases (MMPs) by macrophages is believed to play a key role in matrix degradation underlying plaque instability. Conversely, tissue inhibitors of metalloproteinases (TIMPs) would contribute to plaque stability. We therefore studied the effects of omega-3 fatty acids on the release and activity of MMPs and TIMPs in cultured human monocytoid cells. Methods. Human...

  14. Simultaneous Qualitative Assessment and Quantitative Analysis of Metabolites (Phenolics, Nucleosides and Amino Acids from the Roots of Fresh Gastrodia elata Using UPLC-ESI-Triple Quadrupole Ion MS and ESI- Linear Ion Trap High-Resolution MS.

    Sha Chen

    Full Text Available A sensitive, effective and optimized method, based on ultra performance liquid chromatography (UPLC coupled with ESI-triple quadrupole ion MS and ESI-linear ion trap high-resolution MS, has been developed for the simultaneous quantitative and qualitative determination of phenolics, nucleosides and amino acids in the roots of fresh Gastrodia elata. Optimization of the analytical method provided higher separation efficiency and better peak resolution for the targeted compounds. The simultaneous separation protocols were also optimized by routinely using accurate mass measurements, within 5 ppm error, for each molecular ion and the subsequent fragment ions. In total, 31 compounds, including 23 phenolics, two nucleosides, four amino acids, one gastrodin and one other compound were identified or tentatively characterized. Mono-substituted parishin glucoside (9, methoxy mono-substituted parishin (13, methyl parishin (26, p-hydroxybenzyl di-substituted parishin (29, and p-hydroxybenzyl parishin (31 were tentatively identified as new compounds. Principal metabolite content analysis and the composition of eight representative G. elata cultivars of various species indicated that geographic insulation was the main contributor to clustering.

  15. Simultaneous Qualitative Assessment and Quantitative Analysis of Metabolites (Phenolics, Nucleosides and Amino Acids) from the Roots of Fresh Gastrodia elata Using UPLC-ESI-Triple Quadrupole Ion MS and ESI- Linear Ion Trap High-Resolution MS.

    Chen, Sha; Liu, Jun Qiu; Xiao, Hui; Zhang, Jun; Liu, An

    2016-01-01

    A sensitive, effective and optimized method, based on ultra performance liquid chromatography (UPLC) coupled with ESI-triple quadrupole ion MS and ESI-linear ion trap high-resolution MS, has been developed for the simultaneous quantitative and qualitative determination of phenolics, nucleosides and amino acids in the roots of fresh Gastrodia elata. Optimization of the analytical method provided higher separation efficiency and better peak resolution for the targeted compounds. The simultaneous separation protocols were also optimized by routinely using accurate mass measurements, within 5 ppm error, for each molecular ion and the subsequent fragment ions. In total, 31 compounds, including 23 phenolics, two nucleosides, four amino acids, one gastrodin and one other compound were identified or tentatively characterized. Mono-substituted parishin glucoside (9), methoxy mono-substituted parishin (13), methyl parishin (26), p-hydroxybenzyl di-substituted parishin (29), and p-hydroxybenzyl parishin (31) were tentatively identified as new compounds. Principal metabolite content analysis and the composition of eight representative G. elata cultivars of various species indicated that geographic insulation was the main contributor to clustering. PMID:26954012

  16. Fatty Acids Composition of Vegetable Oils and Its Contribution to Dietary Energy Intake and Dependence of Cardiovascular Mortality on Dietary Intake of Fatty Acids

    Jana Orsavova; Ladislava Misurcova; Jarmila Vavra Ambrozova; Robert Vicha; Jiri Mlcek

    2015-01-01

    Characterizations of fatty acids composition in % of total methylester of fatty acids (FAMEs) of fourteen vegetable oils—safflower, grape, silybum marianum, hemp, sunflower, wheat germ, pumpkin seed, sesame, rice bran, almond, rapeseed, peanut, olive, and coconut oil—were obtained by using gas chromatography (GC). Saturated (SFA), monounsaturated (MUFA) and polyunsaturated fatty acids (PUFA), palmitic acid (C16:0; 4.6%–20.0%), oleic acid (C18:1; 6.2%–71.1%) and linoleic acid (C18:2; 1.6%–79%)...

  17. Contribution of Individual Chemoreceptors to Sinorhizobium meliloti Chemotaxis Towards Amino Acids of Host and Nonhost Seed Exudates.

    Webb, Benjamin A; Helm, Richard F; Scharf, Birgit E

    2016-03-01

    Plant seeds and roots exude a spectrum of molecules into the soil that attract bacteria to the spermosphere and rhizosphere, respectively. The alfalfa symbiont Sinorhizobium meliloti utilizes eight chemoreceptors (McpT to McpZ and IcpA) to mediate chemotaxis. Using a modified hydrogel capillary chemotaxis assay that allows data quantification and larger throughput screening, we defined the role of S. meliloti chemoreceptors in sensing its host, Medicago sativa, and a closely related nonhost, Medicago arabica. S. meliloti wild type and most single-deletion strains displayed comparable chemotaxis responses to host or nonhost seed exudate. However, while the mcpZ mutant responded like wild type to M. sativa exudate, its reaction to M. arabica exudate was reduced by 80%. Even though the amino acid (AA) amounts released by both plant species were similar, synthetic AA mixtures that matched exudate profiles contributed differentially to the S. meliloti wild-type response to M. sativa (23%) and M. arabica (37%) exudates, with McpU identified as the most important chemoreceptor for AA. Our results show that S. meliloti is equally attracted to host and nonhost legumes; however, AA play a greater role in attraction to M. arabica than to M. sativa, with McpZ being specifically important in sensing M. arabica. PMID:26713349

  18. Marine Microbial Secondary Metabolites: Pathways, Evolution and Physiological Roles.

    Giordano, Daniela; Coppola, Daniela; Russo, Roberta; Denaro, Renata; Giuliano, Laura; Lauro, Federico M; di Prisco, Guido; Verde, Cinzia

    2015-01-01

    Microbes produce a huge array of secondary metabolites endowed with important ecological functions. These molecules, which can be catalogued as natural products, have long been exploited in medical fields as antibiotics, anticancer and anti-infective agents. Recent years have seen considerable advances in elucidating natural-product biosynthesis and many drugs used today are natural products or natural-product derivatives. The major contribution to recent knowledge came from application of genomics to secondary metabolism and was facilitated by all relevant genes being organised in a contiguous DNA segment known as gene cluster. Clustering of genes regulating biosynthesis in bacteria is virtually universal. Modular gene clusters can be mixed and matched during evolution to generate structural diversity in natural products. Biosynthesis of many natural products requires the participation of complex molecular machines known as polyketide synthases and non-ribosomal peptide synthetases. Discovery of new evolutionary links between the polyketide synthase and fatty acid synthase pathways may help to understand the selective advantages that led to evolution of secondary-metabolite biosynthesis within bacteria. Secondary metabolites confer selective advantages, either as antibiotics or by providing a chemical language that allows communication among species, with other organisms and their environment. Herewith, we discuss these aspects focusing on the most clinically relevant bioactive molecules, the thiotemplated modular systems that include polyketide synthases, non-ribosomal peptide synthetases and fatty acid synthases. We begin by describing the evolutionary and physiological role of marine natural products, their structural/functional features, mechanisms of action and biosynthesis, then turn to genomic and metagenomic approaches, highlighting how the growing body of information on microbial natural products can be used to address fundamental problems in

  19. Different environmental temperatures affect amino acid metabolism in the eurytherm teleost Senegalese sole (Solea senegalensis Kaup, 1858) as indicated by changes in plasma metabolites.

    Costas, Benjamín; Aragão, Cláudia; Ruiz-Jarabo, Ignacio; Vargas-Chacoff, Luis; Arjona, Francisco J; Mancera, Juan M; Dinis, Maria T; Conceição, Luís E C

    2012-07-01

    Senegalese sole (Solea senegalensis) is a eurytherm teleost that under natural conditions can be exposed to annual water temperature fluctuations between 12 and 26°C. This study assessed the effects of temperature on sole metabolic status, in particular in what concerns plasma free amino acid changes during thermal acclimation. Senegalese sole maintained at 18°C were acclimated to either cold (12°C) or warm (26°C) environmental temperatures for 21 days. Fish maintained at 18°C served as control. Plasma concentrations of cortisol, glucose, lactate, triglycerides, proteins, and free amino acids were assessed. Cold acclimation influenced interrenal responses of sole by increasing cortisol release. Moreover, plasma glucose and lactate concentrations increased linearly with temperature, presumably reflecting a higher metabolic activity of sole acclimated to 26°C. Acclimation temperature affected more drastically plasma concentrations of dispensable than that of indispensable amino acids, and different acclimation temperatures induced different responses. Asparagine, glutamine and ornithine seem to be of particular importance for ammonia detoxification mechanisms, synthesis of triglycerides that may be used during homeoviscous adaptation and, to a lesser extent, as energetic substrates in specimens acclimated to 12°C. When sole is acclimated to 26°C taurine, glutamate, GABA and glycine increased, which may suggest important roles as antioxidant defences, in osmoregulatory processes and/or for energetic purposes at this thermal regimen. In conclusion, acclimation to different environmental temperatures induces several metabolic changes in Senegalese sole, suggesting that amino acids may be important for thermal acclimation. PMID:21947601

  20. Electrogenerated chemiluminescence quenching of Ru(bpy){sub 3} {sup 2+} (bpy=2,2 Prime -bipyridine) in the presence of acetaminophen, salicylic acid and their metabolites

    Haslag, Catherine S. [Department of Chemistry, Missouri State University, Springfield, Missouri 65897 (United States); Richter, Mark M., E-mail: MarkRichter@missouristate.edu [Department of Chemistry, Missouri State University, Springfield, Missouri 65897 (United States)

    2012-03-15

    Quenching of Ru(bpy) {sub 3}{sup 2+} (bpy=2,2 Prime -bipyridine) coreactant electrogenerated chemiluminescence (ECL) has been observed in the presence of acetaminophen, salicylic acid and related complexes. However, no quenching is observed with the acetylsalicylic acid. In most instances, quenching is observed with 100-fold excess of quencher (compared to ECL luminophore) with complete quenching observed between 10,000 and 100,000 fold excess. Fluorescence and UV-vis experiments coupled with bulk electrolysis support the formation of benzoquinone products upon electrochemical oxidation. The mechanism of quenching may involve the interaction of the electrochemically generated benzoquinone species with (i) the {sup Low-Asterisk }Ru(bpy){sub 3}{sup 2+} excited state or (ii) highly energetic coreactant radicals. - Highlights: Black-Right-Pointing-Triangle Efficient quenching of the electrogenerated chemiluminescence is observed. Black-Right-Pointing-Triangle Acetaminophen, salicylic acid and related compounds can be detected. Black-Right-Pointing-Triangle The mechanism of quenching involves benzoquinones formed upon electrolysis.

  1. Effect of dietary sugar concentration and sunflower seed supplementation on lactation performance, ruminal fermentation, milk fatty acid profile, and blood metabolites of dairy cows.

    Razzaghi, A; Valizadeh, R; Naserian, A A; Mesgaran, M Danesh; Carpenter, A J; Ghaffari, M H

    2016-05-01

    Previous research has shown that both sunflower seed (SF) and sucrose (SC) supplementation can result in variation in milk fat concentration and composition, possibly due to altered fermentation patterns and biohydrogenation of fatty acids in the rumen. The objective of this study was to determine the effects of different sugar concentrations with or without SF supplementation on lactation performance, ruminal fermentation, and milk fatty acid profile in lactating dairy cows. Eight multiparous Holstein dairy cows (body weight=620±15kg, 60±10 d in milk, mean ± standard deviation) were randomly assigned to treatments in a replicated 4×4 Latin square design with a 2×2 factorial arrangement of treatments. Each 21-d period consisted of a 14-d diet adaptation period and 7-d collection period. Dairy cows were fed 1 of the following 4 diets: (1) no additional SC without SF supplementation (NSC-SF), (2) no additional SC with SF supplementation (NSC+SF), (3) SC without SF supplementation (SC-SF), and (4) SC with SF supplementation (SC+SF). The diets contained the same amount of forages (corn silage and alfalfa hay). Four isonitrogenous and isoenergetic diets were formulated by replacing corn grain with SC and SF and balanced using change in proportions of canola meal and sugar beet pulp. No interaction was detected between SC and SF supplementation with respect to dry matter intake, milk yield, and composition. A tendency was found for an interaction between inclusion of SC and SF on energy-corrected milk with the highest amount in the SC-SF diet. Ruminal pH and the molar proportion of acetate were affected by SC inclusion, with an increase related to the SC-SF diet. Diets containing SF decreased the concentrations of short-chain fatty acids (4:0 to 10:0) and medium-chain fatty acids (12:0 to 16:0) in milk fat. The addition of SC tended to decrease the concentration of total trans-18:1. These data provide evidence that exchanging SC for corn at 4% of dietary dry matter

  2. Gentisic Acid, a Compound Associated with Plant Defense and a Metabolite of Aspirin, Heads a New Class of in Vivo Fibroblast Growth Factor Inhibitors*

    Fernández, Israel S.; Cuevas, Pedro; Angulo, Javier; López-Navajas, Pilar; Canales-Mayordomo, Ángeles; González-Corrochano, Rocío; Lozano, Rosa M.; Valverde, Serafín; Jiménez-Barbero, Jesús; Romero, Antonio; Giménez-Gallego, Guillermo

    2010-01-01

    Fibroblast growth factors are key proteins in many intercellular signaling networks. They normally remain attached to the extracellular matrix, which confers on them a considerable stability. The unrestrained accumulation of fibroblast growth factors in the extracellular milieu, either due to uncontrolled synthesis or enzymatic release, contributes to the pathology of many diseases. Consequently, the neutralization of improperly mobilized fibroblast growth factors is of clear therapeutic inte...

  3. Metabolic pathways regulated by γ-aminobutyric acid (GABA) contributing to heat tolerance in creeping bentgrass (Agrostis stolonifera).

    Li, Zhou; Yu, Jingjin; Peng, Yan; Huang, Bingru

    2016-01-01

    γ-Aminobutyric acid is a non-protein amino acid involved in various metabolic processes. The objectives of this study were to examine whether increased GABA could improve heat tolerance in cool-season creeping bentgrass through physiological analysis, and to determine major metabolic pathways regulated by GABA through metabolic profiling. Plants were pretreated with 0.5 mM GABA or water before exposed to non-stressed condition (21/19 °C) or heat stress (35/30 °C) in controlled growth chambers for 35 d. The growth and physiological analysis demonstrated that exogenous GABA application significantly improved heat tolerance of creeping bentgrass. Metabolic profiling found that exogenous application of GABA led to increases in accumulations of amino acids (glutamic acid, aspartic acid, alanine, threonine, serine, and valine), organic acids (aconitic acid, malic acid, succinic acid, oxalic acid, and threonic acid), sugars (sucrose, fructose, glucose, galactose, and maltose), and sugar alcohols (mannitol and myo-inositol). These findings suggest that GABA-induced heat tolerance in creeping bentgrass could involve the enhancement of photosynthesis and ascorbate-glutathione cycle, the maintenance of osmotic adjustment, and the increase in GABA shunt. The increased GABA shunt could be the supply of intermediates to feed the tricarboxylic acid cycle of respiration metabolism during a long-term heat stress, thereby maintaining metabolic homeostasis. PMID:27455877

  4. Effects of short-term oilseed supplementation on plasma fatty acid composition, progesterone and prostaglandin F metabolite in lactating beef cows.

    Scholljegerdes, E J; Lekatz, L A; Vonnahme, K A

    2014-05-01

    Twenty-four 3-year-old Angus cows (512.2±21.6 kg) and six ruminally cannulated beef heifers (523.1±16.9 kg) were used to determine the impact of feeding oilseeds starting at the beginning of estrous synchronization until maternal recognition of pregnancy on plasma fatty acid composition. Starting ~60 days postpartum cows were synchronized with the Select Synch+controlled internal drug-release (CIDR) device and timed artificial insemination (AI) protocol. The day CIDR was inserted; cattle were randomly assigned to one of the three treatments being grazing only (CON) or a supplement containing whole soybeans (SOY); or whole flaxseed (FLX). Cattle continued to receive these diets for 28 days. Blood was collected every 3 days until 10 days after insemination and then every day until 18 days after insemination. All cattle grazed a common pasture and supplemented cattle were individually fed their respective supplements once daily. Ruminally cannulated heifers were used to evaluate the impact supplements had on forage intake, which was reduced (P=0.05) with oilseed supplementation. Feeding oilseeds increased total fatty acid intake (Pflax-fed cattle did not exhibit an increase (P=0.02) until day 15 of supplementation over that of CON. Plasma concentrations for 18:3n-3 was greater (Pbreeding had an effect on serum progesterone (P=0.01) with day 16 after timed AI being lower compared with other days. Feeding oilseeds during the time of estrous synchronization will not only increase the energy density of the diet but will provide key fatty acids around the time of maternal recognition of pregnancy. PMID:24572170

  5. Reduced folate and serum vitamin metabolites in patients with rectal carcinoma: an open-label feasibility study of pemetrexed with folic acid and vitamin B12 supplementation.

    Stoffregen, Clemens C; Odin, Elisabeth A; Carlsson, Göran U; Kurlberg, Göran K; Björkqvist, Hillevi G; Tångefjord, Maria T; Gustavsson, Bengt G

    2016-06-01

    The objectives of this single-center, open-label, phase II study were to evaluate (a) the feasibility and safety of neoadjuvant administration of pemetrexed with oral folic acid and vitamin B12 (FA/B12) in newly diagnosed patients with resectable rectal cancer and (b) intracellular and systemic vitamin metabolism. Patients were treated with three cycles of pemetrexed (500 mg/m, every 3 weeks) and FA/B12 before surgery. The reduced folates tetrahydrofolate, 5-methyltetrahydrofolate, and 5,10-methylenetetrahydrofolate were evaluated from biopsies in tumor tissue and in adjacent mucosa. Serum levels of homocysteine, cystathionine, and methylmalonic acid were also measured. All 37 patients received three cycles of pemetrexed; 89.2% completed their planned dosage within a 9-week feasibility time frame. Neither dose reductions nor study drug-related serious adverse events were reported. Reduced folate levels were significantly higher in tumor tissue compared with adjacent mucosa at baseline. After FA/B12 administration, tissue levels of reduced folates increased significantly and remained high during treatment in both tumor and mucosa until surgery. Serum levels of cystathionine increased significantly compared with baseline after FA/B12 administration, but then decreased, fluctuating cyclically during pemetrexed therapy. Homocysteine and methylmalonic acid levels decreased significantly after FA/B12 administration, and remained below baseline levels during the study. These results indicate that administration of three neoadjuvant cycles of single-agent pemetrexed, every 3 weeks, with FA/B12 in patients with resectable rectal cancer is feasible and tolerable. Tissue and serum vitamin metabolism results demonstrate the influence of pemetrexed and FA/B12 on vitamin metabolism and warrant further study. PMID:26825869

  6. Polar metabolites of polycyclic aromatic compounds from fungi are potential soil and groundwater contaminants

    Boll, Esther Sørensen; Johnsen, Anders R.; Christensen, Jan H.

    2015-01-01

    This study investigated the sorption to soil of water-soluble metabolites from polycyclic aromatic compounds (PACs). The soil fungus Cunninghamella elegans was used to produce PAC metabolites from two un-substituted PACs (phenanthrene, pyrene), three alkyl-substituted PACs (2-methylnaphthalene, 1...... calculated for fourteen representative metabolites. Sulfate conjugated metabolites displayed Kd's below 70 whereas the metabolites with both a sulfate and a carboxylic acid group had Kd's below 2.8. The low Kd's of water-soluble PAC metabolites indicate high mobility in soil and a potential for leaching to...

  7. Inhibitory activity of Coprinopsis atramentaria extract, organic acids and their possible methylated synthesized metabolites on growth of human tumor cell lines

    Heleno, Sandrina A.; Ferreira, Isabel C. F. R.; Ricardo C. Calhelha; Martins, Anabela; Queiroz, Maria João R. P.

    2013-01-01

    Coprinopsis atramentaria (Bull.: Fr.) Redhead. Vilgalys & Moncalvo, is a wild edible mushroom previously characterized by our research group for its nutritional composition, and showed a notable antioxidant activity. p-Hydroxybenzoic (4. 71 mg/1 00 g dry weight), p coumaric (0.82 mg/ 100 g) and cinnamic (1.70 mg/100 g) acids were identified in its extract, but have been reported as common compounds in other mushrooms. In the present work, the inhibitory activity of the growth of human t...

  8. Variegatic acid from Serpula lacyrmans reduces FeIII and interacts with other fungal metabolites for location-specific generation and scavenging of reactive oxygen species

    Zhu, Yuan; MAHANEY, JAMES; Jellison, Jody; Cao, Jinzhen; Gressler, Julia; Hoffmeister, Dirk; Goodell, Barry

    2016-01-01

    This study aims to clarify the role of variegatic acid (VA) secreted from Serpula lacyrmans in a chelator-mediated Fenton (CMF) system, including FeIII reduction and the generation of reactive oxygen species (ROS) in the presence of H2O2 and oxalate. As the principle component of the fungal extracellular matrix (ECM), β-glucan isolated from Basidiomycota species was also assessed in scavenging ROS with regard to its potential as a protective barrier for fungal hyphae. Our results demonstrate ...

  9. Genetic variation in primary metabolites of Pastinaca sativa; can herbivores act as selective agents?

    Zangerl, Arthur R; Berenbaum, May R

    2004-10-01

    Although insect herbivory has been shown to act as a selective agent on plant secondary metabolism, whether primary metabolites contribute to resistance and can respond to selection by herbivores remains untested. In the wild parsnip (Pastinaca sativa), its principal herbivore, Depressaria pastinacella, acts as a selective agent on furanocoumarin resistance factors. In this study, we determined whether webworms can, by causing differential reductions in fitness, act as selective agents on parsnip primary metabolites. Estimates of narrow-sense heritabilities were significantly different from zero for C18 fatty acids in buds and developing fruits, fructose and sorbitol in buds, fructose, myo-inositol, bergapten, and psoralen in fruits. Wild parsnips protected from webworms by insecticide produced 2.5 times as much seed biomass as unsprayed plants; that webworms accounted for this difference in plant fitness was indicated by a significant negative relationship between reproductive effort and an index of webworm damage. Only a handful of metabolites influenced resistance to webworms; these included osthol, sorbitol, and protein in developing fruits as well as previously documented furanocoumarins. Osthol, a coumarinic compound, enhanced resistance, as did protein content, while sorbitol lowered resistance. Other primary metabolites may affect resistance to webworms, but their effect was context-dependent, that is, their effect depended on concentrations of other metabolites (epistasis). Susceptible plant phenotypes were found to have average chemical compositions. Although there was genetic variation in some of the primary metabolites in parsnips, the epistatic nature of their involvement in resistance and the lack of genetic variation in some suggest that selection on them from webworms will be either inconsistent or ineffective. PMID:15609832

  10. Propionibacterium acnes CAMP factor and host acid sphingomyelinase contribute to bacterial virulence: potential targets for inflammatory acne treatment.

    Teruaki Nakatsuji

    Full Text Available BACKGROUND: In the progression of acne vulgaris, the disruption of follicular epithelia by an over-growth of Propionibacterium acnes (P. acnes permits the bacteria to spread and become in contact with various skin and immune cells. METHODOLOGY/PRINCIPAL FINDINGS: We have demonstrated in the present study that the Christie, Atkins, Munch-Peterson (CAMP factor of P. acnes is a secretory protein with co-hemolytic activity with sphingomyelinase that can confer cytotoxicity to HaCaT keratinocytes and RAW264.7 macrophages. The CAMP factor from bacteria and acid sphingomyelinase (ASMase from the host cells were simultaneously present in the culture supernatant only when the cells were co-cultured with P. acnes. Either anti-CAMP factor serum or desipramine, a selective ASMase inhibitor, significantly abrogated the P. acnes-induced cell death of HaCaT and RAW264.7 cells. Intradermal injection of ICR mouse ears with live P. acnes induced considerable ear inflammation, macrophage infiltration, and an increase in cellular soluble ASMase. Suppression of ASMase by systemic treatment with desipramine significantly reduced inflammatory reaction induced by intradermal injection with P. acnes, suggesting the contribution of host ASMase in P. acnes-induced inflammatory reaction in vivo. Vaccination of mice with CAMP factor elicited a protective immunity against P. acnes-induced ear inflammation, indicating the involvement of CAMP factor in P. acnes-induced inflammation. Most notably, suppression of both bacterial CAMP factor and host ASMase using vaccination and specific antibody injection, respectively, cooperatively alleviated P. acnes-induced inflammation. CONCLUSIONS/SIGNIFICANCE: These findings envision a novel infectious mechanism by which P. acnes CAMP factor may hijack host ASMase to amplify bacterial virulence to degrade and invade host cells. This work has identified both CAMP factor and ASMase as potential molecular targets for the development of drugs

  11. Differences in urinary trichloroethylene metabolites of animals.

    Ogata,Masana

    1979-12-01

    Full Text Available Differences in urinary excretion of trichloroethylene were studied in rabbits, rats and mice. Trichloretylene (1 m mole/kg was injected intra-peritoneally, then urinary trichloroacetic acid and trichloroethanol glucuronide were measured. The results were: 1. The ratio of total excretion of trichloroethylene metabolites to the administered trichloroethylene decreased in the order of mice, rats and rabbits. 2. The ratio of total trichloroethanol to trichloroacetic acid in urine decreased in the order of rabbits (69.2, mice (12.8 and rats (2.3. The high ratio in rabbits was due to the extremely small amount of trichloroacetic acid in the urine. 3. Differences in these two urinary metabolites in the three kinds of animals and in human subjects were discussed.

  12. Probing the Active Site of MIO-dependent Aminomutases, Key Catalysts in the Biosynthesis of amino Acids Incorporated in Secondary Metabolites

    Cooke, H.; Bruner, S

    2010-01-01

    The tyrosine aminomutase SgTAM produces (S)-{beta}-tyrosine from L-tyrosine in the biosynthesis of the enediyne antitumor antibiotic C-1027. This conversion is promoted by the methylideneimidazole-5-one (MIO) prosthetic group. MIO was first identified in the homologous family of ammonia lyases, which deaminate aromatic amino acids to form {alpha},{beta}-unsaturated carboxylates. Studies of substrate specificity have been described for lyases but there have been limited reports in altering the substrate specificity of aminomutases. Furthermore, it remains unclear as to what structural properties are responsible for catalyzing the presumed readdition of the amino group into the {alpha},{beta}-unsaturated intermediates to form {beta}-amino acids. Attempts to elucidate specificity and mechanistic determinants of SgTAM have also proved to be difficult as it is recalcitrant to perturbations to the active site via mutagenesis. An X-ray cocrystal structure of the SgTAM mutant of the catalytic base with L-tyrosine verified important substrate binding residues as well as the enzymatic base. Further mutagenesis revealed that removal of these crucial interactions renders the enzyme inactive. Proposed structural determinants for mutase activity probed via mutagenesis, time-point assays and X-ray crystallography revealed a complicated role for these residues in maintaining key quaternary structure properties that aid in catalysis.

  13. Probing the active site of MIO-dependent aminomutases, key catalysts in the biosynthesis of beta-amino acids incorporated in secondary metabolites.

    Cooke, Heather A; Bruner, Steven D

    2010-09-01

    The tyrosine aminomutase SgTAM produces (S)-ss-tyrosine from L-tyrosine in the biosynthesis of the enediyne antitumor antibiotic C-1027. This conversion is promoted by the methylideneimidazole-5-one (MIO) prosthetic group. MIO was first identified in the homologous family of ammonia lyases, which deaminate aromatic amino acids to form alpha,ss-unsaturated carboxylates. Studies of substrate specificity have been described for lyases but there have been limited reports in altering the substrate specificity of aminomutases. Furthermore, it remains unclear as to what structural properties are responsible for catalyzing the presumed readdition of the amino group into the alpha,ss-unsaturated intermediates to form ss-amino acids. Attempts to elucidate specificity and mechanistic determinants of SgTAM have also proved to be difficult as it is recalcitrant to perturbations to the active site via mutagenesis. An X-ray cocrystal structure of the SgTAM mutant of the catalytic base with L-tyrosine verified important substrate binding residues as well as the enzymatic base. Further mutagenesis revealed that removal of these crucial interactions renders the enzyme inactive. Proposed structural determinants for mutase activity probed via mutagenesis, time-point assays and X-ray crystallography revealed a complicated role for these residues in maintaining key quaternary structure properties that aid in catalysis. PMID:20577998

  14. INTRACELLULAR ANTIOXIDANT ACTIVITY OF A STREPTOMYCES SP. 8812 SECONDARY METABOLITE, 6,7-DIHYDROXY-3,4-DIHYDROISOQINO- LINE-3-CARBOXYLIC ACID, AND ITS SYNTHETIC DERIVATIVES.

    Guśpiel, Adam; Ziemska, Joanna; Cześcik, Agnieszka; Kawecki, Robert; Solecka, Jolanta

    2016-01-01

    The aim of this study was to determine the antioxidant properties of 6,7-dihydroxy-3,4-dihydroiso- quinoline-3-carboxylic acid (1) and its derivatives in living cells against reactive forms of oxygen and nitrogen, i.e., hydrogen peroxide and nitric oxide. Four of tested compounds scavenged the reactive form of nitrogen more efficiently or similarly to Trolox (EC50 = 55.80 µM). Two compounds exhibited antioxidant activity against reactive oxygen species better than Trolox (EC50 = 51.88 µM). The most active derivative of 1 was the compound containing an iodine atom at position 8 (6,7-dihydroxy-8-iodo-3,4-dihydroisoquinoline-3-carboxylic acid). Our studies showed that some of the derivatives had the ability to cross the cell membrane and scavenge free radicals inside living cells. Thus, they are able to protect DNA and other cellular structures from the dam- aging effects of reactive oxygen and nitrogen species. In addition, some molecular descriptors of the tested compounds were determined with the use of ICM Pro (Molsoft L.L.C.). PMID:27476282

  15. Urinary pesticide metabolites in school students from northern Thailand.

    Panuwet, Parinya; Prapamontol, Tippawan; Chantara, Somporn; Barr, Dana B

    2009-05-01

    We evaluated exposure to pesticides among secondary school students aged 12-13 years old in Chiang Mai Province, Thailand. Pesticide-specific urinary metabolites were used as biomarkers of exposure for a variety of pesticides, including organophosphorus insecticides, synthetic pyrethroid insecticides and selected herbicides. We employed a simple solid-phase extraction with analysis using isotope dilution high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS). A total of 207 urine samples from Thai students were analyzed for 18 specific pesticide metabolites. We found 14 metabolites in the urine samples tested; seven of them were detected with a frequency > or=17%. The most frequently detected metabolites were 2-[(dimethoxyphosphorothioyl) sulfanyl] succinic acid (malathion dicarboxylic acid), para-nitrophenol (PNP), 3,5,6-trichloro-2-pyridinol (TPCY; metabolite of chlorpyrifos), 2,4-dichlorophenoxyacetic acid (2,4-D), cis- and trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carboxylic acids (c-DCCA and t-DCCA; metabolite of permethrin) and 3-phenoxybenzoic acid (3-PBA; metabolite of pyrethroids). The students were classified into 4 groups according to their parental occupations: farmers (N=60), merchants and traders (N=39), government and company employees (N=52), and laborers (N=56). Children of farmers had significantly higher urinary concentrations of pyrethroid insecticide metabolites than did other children (pmetabolite concentrations. Males had significantly higher values of PNP (Mann-Whitney test, p=0.009); however, no other sex-related differences were observed. Because parental occupation and agricultural activities seemed to have little influence on pesticide levels, dietary sources were the likely contributors to the metabolite levels observed. PMID:18760967

  16. Rapid etiological classification of meningitis by NMR spectroscopy based on metabolite profiles and host response.

    Uwe Himmelreich

    Full Text Available Bacterial meningitis is an acute disease with high mortality that is reduced by early treatment. Identification of the causative microorganism by culture is sensitive but slow. Large volumes of cerebrospinal fluid (CSF are required to maximise sensitivity and establish a provisional diagnosis. We have utilised nuclear magnetic resonance (NMR spectroscopy to rapidly characterise the biochemical profile of CSF from normal rats and animals with pneumococcal or cryptococcal meningitis. Use of a miniaturised capillary NMR system overcame limitations caused by small CSF volumes and low metabolite concentrations. The analysis of the complex NMR spectroscopic data by a supervised statistical classification strategy included major, minor and unidentified metabolites. Reproducible spectral profiles were generated within less than three minutes, and revealed differences in the relative amounts of glucose, lactate, citrate, amino acid residues, acetate and polyols in the three groups. Contributions from microbial metabolism and inflammatory cells were evident. The computerised statistical classification strategy is based on both major metabolites and minor, partially unidentified metabolites. This data analysis proved highly specific for diagnosis (100% specificity in the final validation set, provided those with visible blood contamination were excluded from analysis; 6-8% of samples were classified as indeterminate. This proof of principle study suggests that a rapid etiologic diagnosis of meningitis is possible without prior culture. The method can be fully automated and avoids delays due to processing and selective identification of specific pathogens that are inherent in DNA-based techniques.

  17. Enteric bacterial metabolites propionic and butyric acid modulate gene expression, including CREB-dependent catecholaminergic neurotransmission, in PC12 cells--possible relevance to autism spectrum disorders.

    Bistra B Nankova

    Full Text Available Alterations in gut microbiome composition have an emerging role in health and disease including brain function and behavior. Short chain fatty acids (SCFA like propionic (PPA, and butyric acid (BA, which are present in diet and are fermentation products of many gastrointestinal bacteria, are showing increasing importance in host health, but also may be environmental contributors in neurodevelopmental disorders including autism spectrum disorders (ASD. Further to this we have shown SCFA administration to rodents over a variety of routes (intracerebroventricular, subcutaneous, intraperitoneal or developmental time periods can elicit behavioral, electrophysiological, neuropathological and biochemical effects consistent with findings in ASD patients. SCFA are capable of altering host gene expression, partly due to their histone deacetylase inhibitor activity. We have previously shown BA can regulate tyrosine hydroxylase (TH mRNA levels in a PC12 cell model. Since monoamine concentration is known to be elevated in the brain and blood of ASD patients and in many ASD animal models, we hypothesized that SCFA may directly influence brain monoaminergic pathways. When PC12 cells were transiently transfected with plasmids having a luciferase reporter gene under the control of the TH promoter, PPA was found to induce reporter gene activity over a wide concentration range. CREB transcription factor(s was necessary for the transcriptional activation of TH gene by PPA. At lower concentrations PPA also caused accumulation of TH mRNA and protein, indicative of increased cell capacity to produce catecholamines. PPA and BA induced broad alterations in gene expression including neurotransmitter systems, neuronal cell adhesion molecules, inflammation, oxidative stress, lipid metabolism and mitochondrial function, all of which have been implicated in ASD. In conclusion, our data are consistent with a molecular mechanism through which gut related environmental signals

  18. Regulation of Vascular and Renal Function by Metabolite Receptors.

    Peti-Peterdi, János; Kishore, Bellamkonda K; Pluznick, Jennifer L

    2016-01-01

    To maintain metabolic homeostasis, the body must be able to monitor the concentration of a large number of substances, including metabolites, in real time and to use that information to regulate the activities of different metabolic pathways. Such regulation is achieved by the presence of sensors, termed metabolite receptors, in various tissues and cells of the body, which in turn convey the information to appropriate regulatory or positive or negative feedback systems. In this review, we cover the unique roles of metabolite receptors in renal and vascular function. These receptors play a wide variety of important roles in maintaining various aspects of homeostasis-from salt and water balance to metabolism-by sensing metabolites from a wide variety of sources. We discuss the role of metabolite sensors in sensing metabolites generated locally, metabolites generated at distant tissues or organs, or even metabolites generated by resident microbes. Metabolite receptors are also involved in various pathophysiological conditions and are being recognized as potential targets for new drugs. By highlighting three receptor families-(a) citric acid cycle intermediate receptors, (b) purinergic receptors, and PMID:26667077

  19. Effect of L- or DL-methionine Supplementation on Nitrogen Retention, Serum Amino Acid Concentrations and Blood Metabolites Profile in Starter Pigs.

    Tian, Q Y; Zeng, Z K; Zhang, Y X; Long, S F; Piao, X S

    2016-05-01

    The objective of the current study was to evaluate the effect of supplementation of either L-methionine (L-Met) or DL-methionine (DL-Met) to diets of starter pigs on nitrogen (N) balance, metabolism, and serum amino acid profile. Eighteen crossbred (Duroc×Landrace×Yorkshire) barrows weighing 15.45±0.88 kg were randomly allotted to 1 of 3 diets with 6 pigs per treatment. The diets included a basal diet (Met-deficient diet) containing 0.24% standardized ileal digestibility Met with all other essential nutrients meeting the pig's requirements. The other two diets were produced by supplementing the basal diet with 0.12% DL-Met or L-Met. The experiment lasted for 18 days, consisting of a 13-day adaptation period to the diets followed by a 5-day experimental period. Pigs were fed ad libitum and free access to water throughout the experiment. Results showed that the supplementation of either L-Met or DL-Met improved N retention, and serum methionine concentration, and decreased N excretion compared with basal diet (p0.05). In conclusion, on equimolar basis DL-Met and L-Met are equally bioavailable as Met sources for starter pigs. PMID:26954214

  20. Serum bile acids are higher in humans with prior gastric bypass: potential contribution to improved glucose and lipid metabolism

    Patti, Mary-Elizabeth; Houten, Sander M; Bianco, Antonio C;

    2009-01-01

    index cohort (n = 10). Total serum bile acid concentrations were higher in GB (8.90 +/- 4.84 micromol/l) than in both overweight (3.59 +/- 1.95, P = 0.005, Ov) and severely obese (3.86 +/- 1.51, P = 0.045, MOb). Bile acid subfractions taurochenodeoxycholic, taurodeoxycholic, glycocholic...

  1. Colonization of an acid resistant Kingella denitrificans in the stomach may contribute to gastric dysbiosis by Helicobacter pylori.

    Okamoto, Takeshi; Hayashi, Yasuhiro; Mizuno, Hidekazu; Yanai, Hideo; Nishikawa, Jun; Nakazawa, Teruko; Iizasa, Hisashi; Jinushi, Masahisa; Sakaida, Isao; Yoshiyama, Hironori

    2014-03-01

    In the stomach of a gastric ulcer patient who had been administered an anti-acid, a gram-negative and urease-negative bacillus similar in size to Helicobacter pylori was infected together with H. pylori. According to biochemical test and 16S rRNA gene analysis, the urease-negative bacterium was identified as Kingella denitrificans, a human nasopharyngeal commensal. In contrast to the standard strain of K. denitrificans, the isolate showed catalase activity, did not produce acid from glucose, and exhibited acid tolerance. Acid tolerance of H. pylori was increased by cocultivation with the K. denitrificans isolate, but not with other isolates of K. denitrificans. Disruption of physiological and immunological niche by dysbiotic colonization of bacterium may provide pathological attributes to human stomach. Collectively, a careful administration of anti-acids to the elderly, especially those with atrophic gastritis, is necessary to avoid repression of the gastric barrier to bacteria. PMID:24462438

  2. Reactive Metabolites: Current and Emerging Risk and Hazard Assessments.

    Thompson, Richard A; Isin, Emre M; Ogese, Monday O; Mettetal, Jerome T; Williams, Dominic P

    2016-04-18

    Although idiosyncratic adverse drug reactions are rare, they are still a major concern to patient safety. Reactive metabolites are widely accepted as playing a pivotal role in the pathogenesis of idiosyncratic adverse drug reactions. While there are today well established strategies for the risk assessment of stable metabolites within the pharmaceutical industry, there is still no consensus on reactive metabolite risk assessment strategies. This is due to the complexity of the mechanisms of these toxicities as well as the difficulty in identifying and quantifying short-lived reactive intermediates such as reactive metabolites. In this review, reactive metabolite risk and hazard assessment approaches are discussed, and their pros and cons highlighted. We also discuss the nature of idiosyncratic adverse drug reactions, using acetaminophen and nefazodone to exemplify the complexity of the underlying mechanisms of reactive metabolite mediated hepatotoxicity. One of the key gaps moving forward is our understanding of and ability to predict the contribution of immune activation in idiosyncratic adverse drug reactions. Sections are included on the clinical phenotypes of immune mediated idiosyncratic adverse drug reactions and on the present understanding of immune activation by reactive metabolites. The advances being made in microphysiological systems have a great potential to transform our ability to risk assess reactive metabolites, and an overview of the key components of these systems is presented. Finally, the potential impact of systems pharmacology approaches in reactive metabolite risk assessments is highlighted. PMID:26735163

  3. Salivary Metabolite Fingerprint of Type 1 Diabetes in Young Children.

    de Oliveira, Livia Roberta Piedade; Martins, Carla; Fidalgo, Tatiana Kelly Silva; Freitas-Fernandes, Liana Bastos; de Oliveira Torres, Rafaela; Soares, Aline Laignier; Almeida, Fabio C L; Valente, Ana Paula; de Souza, Ivete Pomarico Ribeiro

    2016-08-01

    Metabolomics is an important tool for the evaluation of the human condition, in both health or disease. This study analyzed the salivary components of type I diabetic children (DM1) under six years of age, to assess oral health related to diabetes control, as well as metabolite profiling using NMR. Partial least squared discriminant analysis (PLS-DA) was used to compare healthy (HG) and uncontrolled DM1 subjects that demonstrated a separation between the groups with classificatory performance of ACC = 0.80, R(2) = 0.92, Q(2) = 0.02 and for DM1 children with glycemia >200 mg/dL of ACC = 0.74, R(2) = 0.91, Q(2) = 0.06. The metabolites that mostly contributed to the distinction between the groups in the loading factor were acetate, n-acetyl-sugar, lactate, and sugar. The univariate analysis showed a decreased salivary concentration of succinic acid and increased levels of lactate, acetate, and sucrose in uncontrolled and DM1 children with glycemia >200 mg/dL. The present study demonstrates that the salivary profile of DM1 differs from that of HG children. It appears that diabetes status control has an important effect on the salivary composition. PMID:27306956

  4. Simultaneous determination of mycophenolate mofetil and its active metabolite, mycophenolic acid, by differential pulse voltammetry using multi-walled carbon nanotubes modified glassy carbon electrode

    A highly sensitive electrochemical sensor for the simultaneous determination of mycophenolate mofetil (MPM) and mycophenolic acid (MPA) was fabricated by multi-walled carbon nanotubes modified glassy carbon electrode (MWCNTs/GCE). The electrochemical behavior of these two drugs was studied at the modified electrode using cyclic voltammetry and adsorptive differential pulse voltammetry. MPM and MPA were oxidized at the GCE during an irreversible process. DPV analysis showed two oxidation peaks at 0.87 V and 1.1 V vs. Ag/AgCl for MPM and an oxidation peak at 0.87 V vs. Ag/AgCl for MPA in phosphate buffer solution of pH 5.0. The MWCNTs/GCE displayed excellent electrochemical activities toward oxidation of MPM and MPA relative to the bare GCE. The experimental design algorithm was used for optimization of DPV parameters. The electrode represents linear responses in the range 5.0 × 10−6 to 1.6 × 10−4 mol L−1 and 2.5 × 10−6 mol L−1 to 6.0 × 10−5 mol L−1 for MPM and MPA, respectively. The detection limit was found to be 9.0 × 10−7 mol L−1 and 4.0 × 10−7 mol L−1 for MPM and MPA, respectively. The modified electrode showed a good sensitivity and stability. It was successfully applied to the simultaneous determination of MPM and MPA in plasma and urine samples. - Highlights: • A new modified electrochemical sensor was constructed and used. • Multiwalled carbon nanotubes were used as the modifiers. • MPM and MPA were measured simultaneously at the low levels. • The sensor was used to the determination of MPA and MPM in real samples

  5. Contribution à l'étude de la fonction de transfert air-neige pour les gaz acides (acides organiques, HNO3, HCl)

    Leopold, Anne

    1995-01-01

    Les acides organiques (principalement l'acide formique (HCOOH) et l'acide acétique (CH3COOH) jouent un rôle clef dans la chimie de la troposphère. L'archivage glaciaire représente un moyen privilégié pour obtenir des informations permettant une meilleure compréhension globale du cycle de ces acides carboxyliques. Cependant, la méconnaissance des mécanismes d'incorporation de ces gaz dans la précipitation limite encore l'interprétation de ces archives en terme de variabilité de l'atmosphère pa...

  6. Constitutive Androstane Receptor-Mediated Changes in Bile Acid Composition Contributes to Hepatoprotection from Lithocholic Acid-Induced Liver Injury in MiceS⃞

    Beilke, Lisa D.; Aleksunes, Lauren M.; Holland, Ricky D; Besselsen, David G; Beger, Rick D.; Klaassen, Curtis D.; Cherrington, Nathan J.

    2009-01-01

    Pharmacological activation of the constitutive androstane receptor (CAR) protects the liver during cholestasis. The current study evaluates how activation of CAR influences genes involved in bile acid biosynthesis as a mechanism of hepatoprotection during bile acid-induced liver injury. CAR activators phenobarbital (PB) and 1,4-bis[2-(3,5-dichloropyridyloxy)]benzene (TCPOBOP) or corn oil (CO) were administered to C57BL/6 wild-type (WT) and CAR knockout (CAR-null) mice ...

  7. Contribution of Amino Acid Region 334−335 from Factor Va Heavy Chain to the Catalytic Efficiency of Prothrombinase†

    Barhoover, Melissa A.; Orban, Tivadar; Beck, Daniel O.; Bukys, Michael A.; Kalafatis, Michael

    2008-01-01

    We have demonstrated that amino acids E323, Y324, E330, and V331 from the factor Va heavy chain are required for the interaction of the cofactor with factor Xa and optimum rates of prothrombin cleavage. We have also shown that amino acid region 332−336 contains residues that are important for cofactor function. Using overlapping peptides, we identified amino acids D334 and Y335 as contributors to cofactor activity. We constructed recombinant factor V molecules with the mutations D334 → K and ...

  8. Contribution of Amino Acid Region 659−663 of Factor Va Heavy Chain to the Activity of Factor Xa within Prothrombinase†,‡

    2010-01-01

    Factor Va, the cofactor of prothrombinase, is composed of heavy and light chains associated noncovalently in the presence of divalent metal ions. The COOH-terminal region of the heavy chain contains acidic amino acid clusters that are important for cofactor activity. In this work, we have investigated the role of amino acid region 659−663, which contains five consecutive acidic amino acid residues, by site-directed mutagenesis. We have generated factor V molecules in which all residues were mutated to either lysine (factor V5K) or alanine (factor V5A). We have also constructed a mutant molecule with this region deleted (factor VΔ659−663). The recombinant molecules along with wild-type factor V (factor VWT) were transiently expressed in mammalian cells, purified, and assessed for cofactor activity. Two-stage clotting assays revealed that the mutant molecules had reduced clotting activities compared to that of factor VaWT. Kinetic analyses of prothrombinase assembled with the mutant molecules demonstrated diminished kcat values, while the affinity of all mutant molecules for factor Xa was similar to that for factor VaWT. Gel electrophoresis analyses of plasma-derived and recombinant mutant prothrombin activation demonstrated delayed cleavage of prothrombin at both Arg320 and Arg271 by prothrombinase assembled with the mutant molecules, resulting in meizothrombin lingering throughout the activation process. These results were confirmed after analysis of the cleavage of FPR-meizothrombin. Our findings provide new insights into the structural contribution of the acidic COOH-terminal region of factor Va heavy chain to factor Xa activity within prothrombinase and demonstrate that amino acid region 659−663 from the heavy chain of the cofactor contributes to the regulation of the rate of cleavage of prothrombin by prothrombinase. PMID:20722419

  9. Contribution to the study of the redox couple Np(VI)/Np(V) in the presence of uranium(VI) in solutions of nitric acid and nitrous acid

    The redox behavior of the Np(VI)/Np(V) couple was the subject of a spectrometric study of the Np(VI) reduction reaction in nitric acid solutions (4 to 5 M) containing variable concentrations (1.5 to 3.5 x 10-3 M) of nitrous acid. A low nitrous acid concentration and a high nitric acid concentration were found to favor the stabilization of Np(VI). The stoichiometric coefficients of nitrous acid and nitric acid in the Np(VI) reduction reaction were determined thermodynamically, although only the reaction order with respect to HNO2 could be calculated from a kinetic analysis. Adding nitrate ions to a HNO3/HNO2 solution enhanced the stability of neptunium at oxidation state +VI, but also increased the reduction rate. When uranium(VI) was added to the HNO3/HNO2 solutions, the total quantity of neptunium at oxidation state +V (either free or as a Np(V)-U(VI) complex) remained practically unchanged, as did the Np(VI) reduction rate. The electrochemical behavior of the Np(VI)/Np(V) couple was investigated in a weak acidic medium by voltammetry with an ultra-micro-electrode (UME). The oxidation wave limiting current variation was a linear function of the Np(V) concentration when a gold UME was used, but not with a platinum UME; the reduction wave limiting current variation versus the Np(V) concentration was linear with either gold or platinum UMEs. The presence of the Np(V)-U(VI) complex in the neptunium solutions was characterized by a shift in the normal apparent potential of the Np(VI)/Np(V) couple toward anodic potentials consistent with the previously determined values of the complexation constants. (author)

  10. "ALTERED PLASMA ZINC LEVEL CONTRIBUTES TO THE DEVELOPMENTAL TOXICITY OF VALPROIC ACID IN SKELETAL SYSTEM OF RAT"

    M.Akbari1

    2004-05-01

    Full Text Available Valproic acid is one of the main antiepileptic drugs. There is an increased risk of neural tube defects and axial skeletal malformations among infants born to women who had received valproic acid. There is a hypothesis that one biochemical abnormality underlying the teratogenicity of valproic acid is a drug-induced reduction in maternal plasma zinc .In the present experimental study mated rats were divided into four groups of 8 animals each [control, valproic acid (VPA, valproic acid + zinc (VPA+ Zn and zinc (Zn groups]. The VPA group received 300 mg/kg valproic acid, daily.The control group received an equal volume of 0.9% NaCl. The VPA+ Zn group received 300 mg/kg VPA and 30 mg/kg zinc sulfate and the Zn group received 30 mg/kg zinc sulfate, daily. Valproic acid, NaCl, and Zn were administered intraperitonealy from day 6 through day 15 of gestation. On day 16, six rats of each group were authanized and the other rats were scarified on gestational day (GD 20 to evaluate the skeletal system among the elder fetuses. Blood was drawn to determine plasma zinc. The data were analyzed by using analysis of variance (Kruskal -Wallis test. The zinc concentration in the plasma of rats treated with valproic acid was significantly lower than those of the other groups on 16 GD (P=0.004. Some anomalies such as hydrocephaly, spina bifida, hemivertebrate, and rib malformations were seen in VPA treated group. Low percentage of rib anomalies and spina bifida were observed in the VPA+ Zn treated group while no skeletal anomalies were seen in Zn and control groups. The results from the present experiment support the hypothesis that one of the biochemical abnormalities causing the teratogenicity of VPA is a drug–induced maternal plasma zinc deficiency, and possibly, it may also result in reduction of embryonic Zn.

  11. Three new metabolites from Botrytis cinerea.

    Wang, Tian-Shan; Zhou, Jin-Yan; Tan, Hong

    2008-01-01

    Three new metabolites, gamma-abscisolactone (1), botrytisic acids A (3) and B (4) were isolated from the fermentation broth of Botrytis cinerea TB-3-H8. Their structures were elucidated on the basis of MS, IR, UV, and NMR spectroscopic data. Compound 2 was isolated from natural resource for the first time. The structure of 1 was further confirmed by single-crystal X-ray diffraction (CCDC-265897). PMID:19003608

  12. The selectivity, voltage-dependence and acid sensitivity of the tandem pore potassium channel TASK-1: contributions of the pore domains

    Yuill, KH; Stansfeld, PJ; Ashmole, I; Sutcliffe, MJ; Stanfield, PR

    2007-01-01

    We have investigated the contribution to ionic selectivity of residues in the selectivity filter and pore helices of the P1 and P2 domains in the acid sensitive potassium channel TASK-1. We used site directed mutagenesis and electrophysiological studies, assisted by structural models built through computational methods. We have measured selectivity in channels expressed in Xenopus oocytes, using voltage clamp to measure shifts in reversal potential and current amplitudes when Rb+ or Na+ repla...

  13. Metabolomics for Secondary Metabolite Research

    Eriko Takano

    2013-11-01

    Full Text Available Metabolomics, the global characterization of metabolite profiles, is becoming an increasingly powerful tool for research on secondary metabolite discovery and production. In this review we discuss examples of recent technological advances and biological applications of metabolomics in the search for chemical novelty and the engineered production of bioactive secondary metabolites.

  14. Metabolite Profiling of Induced Mutants of Rice and Soybean

    The low phytic acid (lpa) rice (Os-lpa-XS110-1, Os-lpa-XS110-2) and soybean (Gm-lpa-TW-75-1, Gm-lpa-ZC-2) mutants generated by γ-irradiation were studied, aimed at comparing these mutants to the corresponding wild-types by means of metabolite profiling based on capillary gas chromatography/mass spectrometry. The usefulness of this approach to assist in the elucidation of the types of mutation resulting in reduced contents of phytic acid should be explored. Metabolite profiling aspires to provide a comprehensive picture of the metabolites present in biological systems. It aims at extracting, detecting, identifying, and quantifying a broad spectrum of compounds in a single sample, to provide a deeper insight into complex biological systems. The extraction and fractionation method used allowed a comprehensive coverage of a broad spectrum of low molecular weight metabolites ranging from lipophilic (fatty acids methyl esters, hydrocarbons, free fatty acids, sterols, tocopherols) to hydrophilic (sugars, sugar alcohols, organic acids, amino acids) compounds. For rice, considerable amounts of the peaks detected were statistically significantly different between wild-types and lpa mutants grown in the same field trial. However, only a few of these differences could be consistently observed in all analyzed field trials, indicating a strong influence of the biological variability. Metabolites consistently shown to be significantly different between wild-type and lpa rice mutants, were found to be closely related to the biogenetic pathways leading to phytic acid. This allowed a prediction of the mutation targets for the lpa rice mutants in the biosynthetic pathway of phytic acid. Similar effects, i.e. statistically significantly different levels of metabolites closely related to the biosynthesis of phytic acid, were consistently observed for soybean. (author)

  15. Humic acids and their interactions with metallic elements: Cu II, Eu III, Th IV, U VI: contribution of size exclusion chromatography method and research of complexation models

    The interest given to natural organic matter (humic and fulvic acids) as complexing agents of metallic ions in soils and natural waters becomes more and more important in environmental area. Cation - humic matter interactions have a great importance, a better understanding of the contribution of these substances in natural media specially towards radioactive elements with long life time. Interactions are studied by a chromatographic technique of gel filtration: the dynamic equilibrium method is based on the separation of the formed complex humic macromolecule - metallic ion and the free metallic ion, which due to its size penetrates totally in the pores of the gel. Separation mechanisms of the chromatographic support and the contribution of each parameter, are studied as a function of the buffer nature, its concentration, the PH, the gel porosity and the valence of the metallic cation. This study led to the determination of the appropriate experimental conditions for each cation. A study of metallic binding with humic acid has been undertaken with Cu2+, Eu3+, Th4+, Uo22+. These elements, except copper, have been chosen for their properties similar to the transuranic elements. Different samples of humic acids (commercial, podzolic soil, rendzine soil) are also studied. A deeper research of europium - humic acid interactions by means of different treatment models (discrete or gaussian models) has been undertaken in order to determine the number, the binding site strength and the global interaction constants

  16. Effects of Enhanced UV-B Radiation on Amino Acid and Secondary Metabolite Contents in Mentha piperita L.%UV-B辐射对薄荷药用成分质量分数的影响

    吴能表; 罗红丽; 马红群; 吴督督; 张扬欢; 胡丽涛; 孙金春

    2012-01-01

    To study the effects of UV-B radiation on amino acid and secondary metabolite contents in Mentha piperita L. , M. piperita plants were exposed to enhanced UV-B radiation at 0.15 and 0.35 W/m2 , and flavone, medicinal amino acids, and menthol, menthone, and P-menthone in the volatile oils were determined. Under 0. 15 W/m2 radiation stress, the content of flavone in the plants was significantly increased; the contents of Glu, Lys, Gly and menthol, menthone and P-menthone in the volatile oils were significantly decreased, while Leu and Phe first increased and then decreased, and the content of soluble proteins decreased significantly 20 and 30 days after the treatment. Under 0.15 W/m2 radiation stress, the content of flavone first increased and decreased later and reached the maximum after 30 days; the contents of Glu, Gly, Leu, Tyr, Phe and Arg and the sum of 9 medicinal amino acids were decreased significantly in 10--40 days, the content of soluble proteins was decreased 20 days after the treatment, and the contents of menthol, menthone, P-menthone in the volatile oils were increased significantly. It is concluded from the present work that low dosage UV-B radiation promoted the synthesis of the aromatic amino acid Phe and the branched-chain amino acid Leu while high dosage UV-B radiation decreased the contents of medicinal amino acids of M. piperita L, and that the M. piperita plants could increase the contents of the secondary metabolites such as flavone, menthol, menthone and P-menthone to enhance their adaptability to UV-B radiation.%目的:探讨增强UV-B辐射对胡椒薄荷氨基酸及药用次生代谢物质量分数的影响.方法:人工模拟生长环境中紫外辐射增强处理胡椒薄荷,用分光光度法测胡椒薄荷内黄酮质量分数,氨基酸采用日立L-8800型全自动氨基酸分析仪测定,薄荷挥发油中主要药用成分质量分数测定采用Agilent 6820气相色谱仪(配氢火焰检测器FID,美国安

  17. Contribution of Amino Acid Region 334−335 from Factor Va Heavy Chain to the Catalytic Efficiency of Prothrombinase†

    2008-01-01

    We have demonstrated that amino acids E323, Y324, E330, and V331 from the factor Va heavy chain are required for the interaction of the cofactor with factor Xa and optimum rates of prothrombin cleavage. We have also shown that amino acid region 332−336 contains residues that are important for cofactor function. Using overlapping peptides, we identified amino acids D334 and Y335 as contributors to cofactor activity. We constructed recombinant factor V molecules with the mutations D334 → K and Y335 → F (factor VKF) and D334 → A and Y335 → A (factor VAA). Kinetic studies showed that while factor VaKF and factor VaAA had a KD for factor Xa similar to the KD observed for wild-type factor Va (factor VaWT), the clotting activities of the mutant molecules were impaired and the kcat of prothrombinase assembled with factor VaKF and factor VaAA was reduced. The second-order rate constant of prothrombinase assembled with factor VaKF or factor VaAA for prothrombin activation was ∼10-fold lower than the second-order rate constant for the same reaction catalyzed by prothrombinase assembled with factor VaWT. We also created quadruple mutants combining mutations in the amino acid region 334–335 with mutations at the previously identified amino acids that are important for factor Xa binding (i.e., E323Y324 and E330V331). Prothrombinase assembled with the quadruple mutant molecules displayed a second-order rate constant up to 400-fold lower than the values obtained with prothrombinase assembled with factor VaWT. The data demonstrate that amino acid region 334–335 is required for the rearrangement of enzyme and substrate necessary for efficient catalysis of prothrombin by prothrombinase. PMID:18537263

  18. Protein adduct formation by glucuronide metabolites of permethrin

    Noort, D.; Zuylen, A. van; Fidder, A.; Ommen, B. van; Hulst, A.G.

    2008-01-01

    Biomonitoring of exposure to the insecticide permethrin is usually performed by analysis of its urinary metabolites 3-phenoxybenzoic acid (3-PBA) or cis/trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carboxylic acid (Cl2CA). We are engaged in the development of a methodology to assess the cu

  19. Metabolite profiling of induced mutants of rice and soybean

    The study objects of the investigation were two low phytic acid (lpa) rice (Os-lpa-XS110-1, Os-lpa-XS110-2) and soybean (Gm-lpa-TW-75-1, Gm-lpa-ZC-2) mutants generated by irradiation. The aim was to compare these mutants to the corresponding wild-types by means of capillary gas chromatography metabolite profiling and to explore the usefulness of this approach to assist in the elucidation of the types of mutation resulting in the reduced contents of phytic acid. Metabolite profiling aspires to provide a comprehensive picture of the metabolites present in biological systems. It aims at extracting, detecting, identifying, and quantifying a broad spectrum of compounds in a single sample to provide a deeper insight into complex biological systems. The extraction and fractionation method used in the study allowed a comprehensive coverage of a broad spectrum of low molecular weight metabolites ranging from lipophilic (fatty acids methyl esters, hydrocarbons, free fatty acids, sterols, tocopherols) to hydrophilic (sugars, sugar alcohols, organic acids, amino acids) compounds. For rice, considerable amounts of the peaks detected were statistically significantly different between wild-types and lpa mutants within one field trial. However, only a few of these differences could be consistently observed in all analyzed field trials indicating a strong influence of the biological variability. Metabolites shown to be consistently statistically significantly different between wild-type and lpa rice mutants were found to be closely related to the biogenetic pathways leading to phytic acid. This allowed a prediction of the mutation targets for the lpa rice mutants in the biosynthetic pathway of phytic acid. Similar effects, e.g. clustering of wild-types and lpa mutants on the basis of metabolite profiling data, were observed for soybean. (author)

  20. Growth promoting effects of some lichen metabolites on probiotic bacteria.

    Gaikwad, Subhash; Verma, Neeraj; Sharma, B O; Behera, B C

    2014-10-01

    In the present study, the extract of four natural lichen species Canoparmelia eruptens, Everniastrum cirrhatum, Parmotrema austrosinense and Rimelia cetrata were studied for the source of natural antioxidant and their purified secondary metabolites were evaluated for growth promoting effects on probiotic bacteria Lactobacillus casei. The methanolic fraction of lichen species showed moderate to high antioxidant activity in the order P. austrosinense > E. cirrhatum > C. eruptens > R. cetrata. The lichen metabolites showed antioxidant activity with an IC50 values (μg/ml); lecanoric acid 79-95, salazinic 88-108, atranorin 100-116 and consalazinic acid 119-125. As far as the growth promoting effects of lichen metabolites on L. casei is concerned, lecanoric acid at 100 μg/ml conc. showed high growth stimulating activity in terms of increased dry matter of biomass (56.08 mg) of L. casei. Other lichen metabolites; salazinic acid, atranorin and consalazinic acid produced relatively less dry biomass 43.98 mg, 41.1 mg, 40.68 mg, respectively. However, standard antioxidants butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT) and Trolox after 36 h produced 39.04-47.81 mg dry biomass. At lower pH the growth promoting activity of lichen metabolites was found stable. PMID:25328204

  1. Salicylic acid and cysteine contribute to arbutin-induced alleviation of angular leaf spot disease development in cucumber.

    Kuźniak, Elżbieta; Wielanek, Marzena; Chwatko, Grażyna; Głowacki, Rafał; Libik-Konieczny, Marta; Piątek, Milena; Gajewska, Ewa; Skłodowska, Maria

    2015-06-01

    Arbutin induced suppression of angular leaf spot disease in cucumber resulting from lower populations of Pseudomonas syringae pv lachrymans in the infected tissues. This study provides insight into mechanisms that may potentially account for this effect. In the absence of the pathogen, exogenous arbutin-induced expression of PR1, the marker of salicylic acid signaling, increased the content of salicylic acid and modulated the cysteine pool. This suggested that arbutin promoted cucumber plants to a "primed" state. When challenged with the pathogen, the arbutin-treated plants showed strongly reduced infection symptoms 7 days after inoculation. At this time point, they were characterized by higher contents of free and protein-bound cysteine due to higher cysteine biosynthetic capacity related to increased activities of serine acetyltransferase and cysteine synthase when compared with plants infected without arbutin treatment. Moreover, in the arbutin-treated and infected plants the contents of free salicylic acid and its conjugates were also increased, partly owing to its biosynthesis via the phenylpropanoid pathway. We suggest that arbutin-induced abrogation of angular leaf spot disease in cucumber could be mediated by salicylic acid and cysteine-based signaling. PMID:25955697

  2. Cystathionine γ-lyase, a H2S-generating enzyme, is a GPBAR1-regulated gene and contributes to vasodilation caused by secondary bile acids.

    Renga, Barbara; Bucci, Mariarosaria; Cipriani, Sabrina; Carino, Adriana; Monti, Maria Chiara; Zampella, Angela; Gargiulo, Antonella; d'Emmanuele di Villa Bianca, Roberta; Distrutti, Eleonora; Fiorucci, Stefano

    2015-07-01

    GPBAR1 is a bile acid-activated receptor (BAR) for secondary bile acids, lithocholic (LCA) and deoxycholic acid (DCA), expressed in the enterohepatic tissues and in the vasculature by endothelial and smooth muscle cells. Despite that bile acids cause vasodilation, it is unclear why these effects involve GPBAR1, and the vascular phenotype of GPBAR1 deficient mice remains poorly defined. Previous studies have suggested a role for nitric oxide (NO) in regulatory activity exerted by GPBAR1 in liver endothelial cells. Hydrogen sulfide (H2S) is a vasodilatory agent generated in endothelial cells by cystathionine-γ-lyase (CSE). Here we demonstrate that GPBAR1 null mice had increased levels of primary and secondary bile acids and impaired vasoconstriction to phenylephrine. In aortic ring preparations, vasodilation caused by chenodeoxycholic acid (CDCA), a weak GPBAR1 ligand and farnesoid-x-receptor agonist (FXR), was iberiotoxin-dependent and GPBAR1-independent. In contrast, vasodilation caused by LCA was GPBAR1 dependent and abrogated by propargyl-glycine, a CSE inhibitor, and by 5β-cholanic acid, a GPBAR1 antagonist, but not by N(5)-(1-iminoethyl)-l-ornithine (l-NIO), an endothelial NO synthase inhibitor, or iberiotoxin, a large-conductance calcium-activated potassium (BKCa) channels antagonist. In venular and aortic endothelial (HUVEC and HAEC) cells GPBAR1 activation increases CSE expression/activity and H2S production. Two cAMP response element binding protein (CREB) sites (CREs) were identified in the CSE promoter. In addition, TLCA stimulates CSE phosphorylation on serine residues. In conclusion we demonstrate that GPBAR1 mediates the vasodilatory activity of LCA and regulates the expression/activity of CSE. Vasodilation caused by CDCA involves BKCa channels. The GPBAR1/CSE pathway might contribute to endothelial dysfunction and hyperdynamic circulation in liver cirrhosis. PMID:25934094

  3. The cadmium–mercaptoacetic acid complex contributes to the genotoxicity of mercaptoacetic acid-coated CdSe-core quantum dots

    Tang, Weikun; Fan, Junpeng; He, Yide; Huang, Bihai; Liu, Huihui; Pang, Daiwen; Xie, Zhixiong

    2012-01-01

    Quantum dots (QDs) have many potential clinical and biological applications because of their advantages over traditional fluorescent dyes. However, the genotoxicity potential of QDs still remains unclear. In this paper, a plasmid-based system was designed to explore the genotoxic mechanism of QDs by detecting changes in DNA configuration and biological activities. The direct chemicobiological interactions between DNA and mercaptoacetic acid-coated CdSecore QDs (MAA–QDs) were investigated. Aft...

  4. Acidic leucine-rich nuclear phosphoprotein 32 family member B (ANP32B) contributes to retinoic acid-induced differentiation of leukemic cells

    Yu, Yun; Shen, Shao-Ming; Zhang, Fei-Fei; Wu, Zhao-Xia; Han, Bin [Shanghai Universities E-Institute for Chemical Biology, Key Laboratory of Cell Differentiation and Apoptosis of National Ministry of Education, Rui-Jin Hospital, Shanghai Jiao-Tong University School of Medicine (SJTU-SM), Shanghai 200025 (China); Wang, Li-Shun, E-mail: jywangls@shsmu.edu.cn [Shanghai Universities E-Institute for Chemical Biology, Key Laboratory of Cell Differentiation and Apoptosis of National Ministry of Education, Rui-Jin Hospital, Shanghai Jiao-Tong University School of Medicine (SJTU-SM), Shanghai 200025 (China)

    2012-07-13

    Highlights: Black-Right-Pointing-Pointer ANP32B was down-regulated during ATRA-induced leukemic cell differentiation. Black-Right-Pointing-Pointer Knockdown of ANP32B enhanced ATRA-induced leukemic cell differentiation. Black-Right-Pointing-Pointer Ectopic expression of ANP32B inhibited ATRA-induced leukemic cell differentiation. Black-Right-Pointing-Pointer ANP32B inhibited ATRA activated transcriptional activity of RAR{alpha}. -- Abstract: The acidic leucine-rich nuclear phosphoprotein 32B (ANP32B) is a member of a conserved superfamily of nuclear proteins whose functions are largely unknown. In our previous work, ANP32B was identified as a novel direct substrate for caspase-3 and acted as a negative regulator for leukemic cell apoptosis. In this work, we provided the first demonstration that ANP32B expression was down-regulated during differentiation induction of leukemic cells by all-trans retinoic acid (ATRA). Knockdown of ANP32B expression by specific shRNA enhanced ATRA-induced leukemic cell differentiation, while ectopic expression of ANP32B attenuated it, indicating an inhibitory role of ANP32B against leukemic cell differentiation. Furthermore, luciferase reporter assay revealed that ANP32B might exert this role through inhibiting the ATRA dependent transcriptional activity of retinoic acid receptor (RAR{alpha}). These data will shed new insights into understanding the biological functions of ANP32B protein.

  5. Acidic leucine-rich nuclear phosphoprotein 32 family member B (ANP32B) contributes to retinoic acid-induced differentiation of leukemic cells

    Highlights: ► ANP32B was down-regulated during ATRA-induced leukemic cell differentiation. ► Knockdown of ANP32B enhanced ATRA-induced leukemic cell differentiation. ► Ectopic expression of ANP32B inhibited ATRA-induced leukemic cell differentiation. ► ANP32B inhibited ATRA activated transcriptional activity of RARα. -- Abstract: The acidic leucine-rich nuclear phosphoprotein 32B (ANP32B) is a member of a conserved superfamily of nuclear proteins whose functions are largely unknown. In our previous work, ANP32B was identified as a novel direct substrate for caspase-3 and acted as a negative regulator for leukemic cell apoptosis. In this work, we provided the first demonstration that ANP32B expression was down-regulated during differentiation induction of leukemic cells by all-trans retinoic acid (ATRA). Knockdown of ANP32B expression by specific shRNA enhanced ATRA-induced leukemic cell differentiation, while ectopic expression of ANP32B attenuated it, indicating an inhibitory role of ANP32B against leukemic cell differentiation. Furthermore, luciferase reporter assay revealed that ANP32B might exert this role through inhibiting the ATRA dependent transcriptional activity of retinoic acid receptor (RARα). These data will shed new insights into understanding the biological functions of ANP32B protein.

  6. Contribution to the study of the physico-chemical mechanisms of metallic cation extraction by alkylphosphoric acids. Extraction of zirconium (IV) by di-2-ethylhexyl phosphoric acid (DEHPA)

    Extraction of zirconium, especially at high concentration (0.1M), by dodecane diluted DEHPA (HA) from hydrochloric or nitric aqueous phases of 0.1 to 10 M acidity was studied. The composition, structure and polymerisation of the complexes extracted were determined by chemical analysis, viscosimetry, infrared spectrometry and light scattering. A Zr(OH)2A2.2HNO3, type structure is proposed for these complexes instead of the generally accepted form: Zr(OH)2(NO3)2.2HA. Similarly in hydrochloric solution: Zr(OH)2A2.2HCl. Polymerisation in the organic phase results from the juxtaposition of two factors; firstly zirconium saturation (formation of bridges by DEHPA between zirconium atoms) and secondly the nature the equeous phase. In slightly acid hydrochloric solution (pH = 1.3) the aqueous plymers of zirconium are extracted in the organic phase as polynuclear complexes; in nitric solution no polynuclear complexes are observed but the nitric acid molecules extracted set up hydrogen bonds which explain the increased viscosity and gelification of the organic phases

  7. Intracellular Metabolite Pool Changes in Response to Nutrient Depletion Induced Metabolic Switching in Streptomyces coelicolor

    Alexander Wentzel

    2012-02-01

    Full Text Available A metabolite profiling study of the antibiotic producing bacterium Streptomyces coelicolor A3(2 has been performed. The aim of this study was to monitor intracellular metabolite pool changes occurring as strains of S. coelicolor react to nutrient depletion with metabolic re-modeling, so-called metabolic switching, and transition from growth to secondary metabolite production phase. Two different culture media were applied, providing depletion of the key nutrients phosphate and L-glutamate, respectively, as the triggers for metabolic switching. Targeted GC-MS and LC-MS methods were employed to quantify important primary metabolite groups like amino acids, organic acids, sugar phosphates and other phosphorylated metabolites, and nucleotides in time-course samples withdrawn from fully-controlled batch fermentations. A general decline, starting already in the early growth phase, was observed for nucleotide pools and phosphorylated metabolite pools for both the phosphate and glutamate limited cultures. The change in amino acid and organic acid pools were more scattered, especially in the phosphate limited situation while a general decrease in amino acid and non-amino organic acid pools was observed in the L-glutamate limited situation. A phoP deletion mutant showed basically the same metabolite pool changes as the wild-type strain M145 when cultivated on phosphate limited medium. This implies that the inactivation of the phoP gene has only little effect on the detected metabolite levels in the cell. The energy charge was found to be relatively constant during growth, transition and secondary metabolite production phase. The results of this study and the employed targeted metabolite profiling methodology are directly relevant for the evaluation of precursor metabolite and energy supply for both natural and heterologous production of secondary metabolites in S. coelicolor.

  8. Assessing the Fatty Acid, Carotenoid, and Tocopherol Compositions of Amaranth and Quinoa Seeds Grown in Ontario and Their Overall Contribution to Nutritional Quality.

    Tang, Yao; Li, Xihong; Chen, Peter X; Zhang, Bing; Liu, Ronghua; Hernandez, Marta; Draves, Jamie; Marcone, Massimo F; Tsao, Rong

    2016-02-10

    Various fatty acids, tocopherols, carotenoids, and their respective antioxidant contributions in 7 amaranth seed and 11 quinoa seed samples along with a new evaluation method are reported. The lipid yield was 6.98-7.22% in amaranth seeds and 6.03-6.74% in quinoa seeds, with unsaturated fatty acids (UFAs) being the predominant fatty acids, 71.58-72.44% in amaranth seeds and 81.44-84.49% in quinoa seeds, respectively. Carotenoids, mainly lutein and zeaxanthin, are confirmed for the first time in amaranth seeds, while β-carotene is reported first in quinoa seeds. The predominant tocopherols in amaranth seeds are δ- and α-tocopherol, whereas γ- and α-tocopherol are the primary tocopherols in quinoa seeds. UFAs, carotenoids, and tocopherols showed good correlation with antioxidant activity. All of the amaranth seeds demonstrated lower overall lipophilic quality than quinoa seeds, with the AS1 and QS10 cultivars providing the highest scores for amaranth and quinoa seeds, respectively. Results from this study will contribute to developing quinoa seeds and related functional foods with increased benefits. PMID:26760897

  9. Amino acid biogeochemistry and bacterial contribution to sediment organic matter along the western margin of the Bay of Bengal

    Fernandes, L.; Garg, A.; Borole, D.V.

    influence burial of organic carbon (Cowie and Hedges, 1992; Burdige, 2007). OM buried in the marine sediments forms a major link between the "active" surface pools of carbon and inactive and/or slow cycling carbon pools (Burdige, 2007). In the open ocean... preserved in the marine sediments (Keil et al., 2000; Vandewiele et al., 2009). Moreover, natural occurrence and geochemical behavior of amino acids have been evaluated in several types of samples (Cowie and Hedges, 1992; Gupta and Kawahata, 2000...

  10. Light-absorbing aldol condensation products in acidic aerosols: Spectra, kinetics, and contribution to the absorption index

    Nozière, Barbara; Esteve, William

    The radiative properties of aerosols that are transparent to light in the near-UV and visible, such as sulfate aerosols, can be dramatically modified when mixed with absorbing material such as soot. In a previous work we had shown that the aldol condensation of carbonyl compounds produces light-absorbing compounds in sulfuric acid solutions. In this work we report the spectroscopic and kinetic parameters necessary to estimate the effects of these reactions on the absorption index of sulfuric acid aerosols in the atmosphere. The absorption spectra obtained from the reactions of six different carbonyl compounds (acetaldehyde, acetone, propanal, butanal, 2-butanone, and trifluoroacetone) and their mixtures were compared over 190-1100 nm. The results indicated that most carbonyl compounds should be able to undergo aldol condensation. The products are oligomers absorbing light in the 300-500 nm region where few other compounds absorb, making them important for the radiative properties of aerosols. Kinetic experiments in 96-75 wt% H 2SO 4 solutions and between 273 and 314 K gave an activation energy for the rate constant of formation of the aldol products of acetaldehyde of -(70±15) kJ mol -1 in 96 wt% solution and showed that the effect of acid concentration was exponential. A complete expression for this rate constant is proposed where the absolute value in 96 wt% H 2SO 4 and at 298 K is scaled to the Henry's law coefficient for acetaldehyde and the absorption cross-section for the aldol products assumed in this work. The absorption index of stratospheric sulfuric acid aerosols after a 2-year residence time was estimated to 2×10 -4, optically equivalent to a content of 0.5% of soot and potentially significant for the radiative forcing of these aerosols and for satellite observations in channels where the aldol products absorb.

  11. Quantification of sarin and cyclosarin metabolites isopropyl methylphosphonic acid and cyclohexyl methylphosphonic acid in minipig plasma using isotope-dilution and liquid chromatography- time-of-flight mass spectrometry.

    Evans, R A; Jakubowski, E M; Muse, W T; Matson, K; Hulet, S W; Mioduszewski, R J; Thomson, S A; Totura, A L; Renner, J A; Crouse, C L

    2008-01-01

    An analysis method for determining isopropyl methylphosphonic acid (IMPA) and cyclohexyl methylphosphonic acid (CMPA), the metabolic hydrolysis products of toxic organophosphorus nerve agents isopropyl methylphosphonofluoridate (sarin, GB) and cyclohexyl methylphosphonofluoridate (cyclosarin, GF), respectively, has been developed and validated using high-performance liquid chromatography-mass spectrometry with negative ion electrospray ionization with time-of-flight detection (LC-ESI-MS-TOF). The linear range of quantitation was 5 to 125 ng/mL in plasma with a method detection limit of 2 ng/mL for each compound. This method was developed to determine the amount of metabolic hydrolysis that was formed during and after nerve agent exposure in minipigs to account for a major pathway of GB and GF elimination that had not been previously characterized in the bloodstream, particularly during low-level whole-body inhalation experiments. Metabolic hydrolysis accounted for 70% to 90% of the recoverable agent in the bloodstream during exposure, when compared to both unbound and cholinesterase bound agent recovered by fluoride ion reactivation analysis for the same samples. The estimated half-life of IMPA and CMPA in plasma was determined to be 44 and 61 min, respectively. The method utilizes the mass selectivity of LC-ESI-MS-TOF using a bench-top instrument to achieve a detection limit that is consistent with reported LC-MS-MS methods analyzing blood samples. PMID:18269798

  12. [Metabolism of etofenamate / Identification and analytic of metabolites, their pharmacological properties and species dependence of metabolism (author's transl)].

    Dell, H D; Doersing, M; Fischer, W; Fiedler, J; Jacobi, H; Kamp, R

    1981-01-01

    After oral application of etofenamate to animals (rat, rabbit, dog, monkey) unchanged etofenamate and numerous metabolites are found in urine. Analytical properties (thin-layer chromatographic behavior, UV- and fluorescence data) of etofenamate, 5-hydroxy-, 4'-hydroxy, 5,4'-dihydroxy-etofenamate, flufenamic acid, 5-hydroxy-, 4'-hydroxy-, 5,4'-dihydroxy-flufenamic acid are described. Derivatives of etofenamate and flufenamic acid are excreted by rabbit, dog, monkey and man, whereas flufenamic acid derivatives are excreted preferentially by rats; profound degradation takes place in dogs. Metabolism in man is more similar to monkey than to dog and rodents. Metabolic pattern after oral and cutaneous application is quite similar. The six hydroxy derivatives have no pharmacological activity--they do not contribute to the pharmacological action of the substance. PMID:6971113

  13. Production of Metabolites

    2011-01-01

    A recombinant micro-organism such as Saccharomyces cerevisiae which produces and excretes into culture medium a stilbenoid metabolite product when grown under stilbenoid production conditions, which expresses in above native levels a ABC transporter which transports said stilbenoid out of said...... micro-organism cells to the culture medium. The genome of the Saccharomyces cerevisiae produces an auxotrophic phenotype which is compensated by a plasmid which also expresses one or more of said enzymes constituting said metabolic pathway producing said stilbenoid, an expression product of the plasmid...

  14. De novo fatty acid biosynthesis contributes significantly to establishment of a bioenergetically favorable environment for vaccinia virus infection.

    Matthew D Greseth

    2014-03-01

    Full Text Available The poxvirus life cycle, although physically autonomous from the host nucleus, is nevertheless dependent upon cellular functions. A requirement for de novo fatty acid biosynthesis was implied by our previous demonstration that cerulenin, a fatty acid synthase inhibitor, impaired vaccinia virus production. Here we show that additional inhibitors of this pathway, TOFA and C75, reduce viral yield significantly, with partial rescue provided by exogenous palmitate, the pathway's end-product. Palmitate's major role during infection is not for phospholipid synthesis or protein palmitoylation. Instead, the mitochondrial import and β-oxidation of palmitate are essential, as shown by the impact of etomoxir and trimetazidine, which target these two processes respectively. Moreover, the impact of these inhibitors is exacerbated in the absence of exogenous glucose, which is otherwise dispensable for infection. In contrast to glucose, glutamine is essential for productive viral infection, providing intermediates that sustain the TCA cycle (anaplerosis. Cumulatively, these data suggest that productive infection requires the mitochondrial β-oxidation of palmitate which drives the TCA cycle and energy production. Additionally, infection causes a significant rise in the cellular oxygen consumption rate (ATP synthesis that is ablated by etomoxir. The biochemical progression of the vaccinia life cycle is not impaired in the presence of TOFA, C75, or etomoxir, although the levels of viral DNA and proteins synthesized are somewhat diminished. However, by reversibly arresting infections at the onset of morphogenesis, and then monitoring virus production after release of the block, we determined that virion assembly is highly sensitive to TOFA and C75. Electron microscopic analysis of cells released into C75 revealed fragmented aggregates of viroplasm which failed to be enclosed by developing virion membranes. Taken together, these data indicate that vaccinia

  15. De novo fatty acid biosynthesis contributes significantly to establishment of a bioenergetically favorable environment for vaccinia virus infection.

    Greseth, Matthew D; Traktman, Paula

    2014-03-01

    The poxvirus life cycle, although physically autonomous from the host nucleus, is nevertheless dependent upon cellular functions. A requirement for de novo fatty acid biosynthesis was implied by our previous demonstration that cerulenin, a fatty acid synthase inhibitor, impaired vaccinia virus production. Here we show that additional inhibitors of this pathway, TOFA and C75, reduce viral yield significantly, with partial rescue provided by exogenous palmitate, the pathway's end-product. Palmitate's major role during infection is not for phospholipid synthesis or protein palmitoylation. Instead, the mitochondrial import and β-oxidation of palmitate are essential, as shown by the impact of etomoxir and trimetazidine, which target these two processes respectively. Moreover, the impact of these inhibitors is exacerbated in the absence of exogenous glucose, which is otherwise dispensable for infection. In contrast to glucose, glutamine is essential for productive viral infection, providing intermediates that sustain the TCA cycle (anaplerosis). Cumulatively, these data suggest that productive infection requires the mitochondrial β-oxidation of palmitate which drives the TCA cycle and energy production. Additionally, infection causes a significant rise in the cellular oxygen consumption rate (ATP synthesis) that is ablated by etomoxir. The biochemical progression of the vaccinia life cycle is not impaired in the presence of TOFA, C75, or etomoxir, although the levels of viral DNA and proteins synthesized are somewhat diminished. However, by reversibly arresting infections at the onset of morphogenesis, and then monitoring virus production after release of the block, we determined that virion assembly is highly sensitive to TOFA and C75. Electron microscopic analysis of cells released into C75 revealed fragmented aggregates of viroplasm which failed to be enclosed by developing virion membranes. Taken together, these data indicate that vaccinia infection, and in

  16. The combined treatment of the intrinsic and external contributions to the stability of the nucleic acids containing unnatural nucleosides

    Czernek, Jiří

    Bucharest: World Scientific and Engineerin Academy and Society Press, 2008 - (Iliescu, M.; Munteanu, R.; Frausto-Solis, J.), s. 228-232 ISBN 978-960-474-019-2. [WSEAS International Conference on Mathematical and Computational Methods in Science and Engineering /10./. Bucharest (RO), 07.11.2008-09.11.2008] R&D Projects: GA AV ČR KJB400500602 Institutional research plan: CEZ:AV0Z40500505 Keywords : nucleic acid * nucleobase * hydrogen-bonding * stacking Subject RIV: CD - Macromolecular Chemistry http://www.worldses.org/review/index.html

  17. Single substitutions to closely related amino acids contribute to the functional diversification of an insect-inducible, positively selected plant cystatin.

    Rasoolizadeh, Asieh; Goulet, Marie-Claire; Sainsbury, Frank; Cloutier, Conrad; Michaud, Dominique

    2016-04-01

    A causal link has been reported between positively selected amino acids in plant cystatins and the inhibitory range of these proteins against insect digestive cysteine (Cys) proteases. Here we assessed the impact of single substitutions to closely related amino acids on the contribution of positive selection to cystatin diversification. Cystatin sequence alignments, while confirming hypervariability, indicated a preference for related amino acids at positively selected sites. For example, the non-polar residues leucine (Leu), isoleucine (Ile) and valine (Val) were shown to predominate at positively selected site 2 in the N-terminal region, unlike selected sites 6 and 10, where polar residues are preferred. The model cystatin SlCYS8 and single variants with Leu, Ile or Val at position 2 were compared with regard to their ability to bind digestive proteases of the coleopteran pest Leptinotarsa decemlineata and to induce compensatory responses in this insect. A functional proteomics procedure to capture target Cys proteases in midgut extracts allowed confirmation of distinct binding profiles for the cystatin variants. A shotgun proteomics procedure to monitor whole Cys protease complements revealed protease family specific compensatory responses in the insect, dependent on the variant ingested. Our data confirm the contribution of closely related amino acids to the functional diversity of positively selected plant cystatins in a broader structure/function context imposing physicochemical constraints to primary structure alterations. They also underline the complexity of protease/inhibitor interactions in plant-insect systems, and the challenges still to be met in order to harness the full potential of ectopically expressed protease inhibitors in crop protection. PMID:26833679

  18. Hydrological modeling of a watershed affected by acid mine drainage (Odiel River, SW Spain). Assessment of the pollutant contributing areas

    Galván, L.; Olías, M.; Cánovas, C. R.; Sarmiento, A. M.; Nieto, J. M.

    2016-09-01

    The Odiel watershed drains materials belonging to the Iberian Pyrite Belt, where significant massive sulfide deposits have been mined historically. As a result, a huge amount of sulfide-rich wastes are deposited in the watershed, which suffer from oxidation, releasing acidic lixiviates with high sulfate and metal concentrations. In order to reliably estimate the metal loadings along the watershed a complete series of discharge and hydrochemical data are essential. A hydrological model was performed with SWAT (Soil and Water Assessment Tool) to solve the scarcity of gauge stations along the watershed. The model was calibrated and validated from daily discharge data (from 1980 to 2010) at the outlet of the watershed, river inputs into an existent reservoir, and a flow gauge station close to the northern area of the watershed. Discharge data obtained from the hydrological model, together with analytical data, allowed the estimation of the dissolved pollutant load delivered annually by the Odiel River (e.g. 9140 t of Al, 2760 t of Zn). The pollutant load is influenced strongly by the rainfall regime, and can even double during extremely rainy years. Around 50% of total pollution comes from the Riotinto Mining District, so the treatment of Riotinto lixiviates reaching the Odiel watershed would reduce the AMD (Acid Mine Drainages) in a remarkable way, improving the water quality downstream, especially in the reservoir of Alcolea, currently under construction. The information obtained in this study will allow the optimization of remediation efforts in the watershed, in order to improve its water quality.

  19. [Antiviral properties of basidiomycetes metabolites].

    Avtonomova, A V; Krasnopolskaya, L M

    2014-01-01

    The data on the antiviral action of the Ganoderma lucidum, Lentinus edodes, Grifola frondosa, Agaricus brasiliensis and other basidiomycetes metabolites are summurized. The metabolites of these species of basidiomycetes exhibit a direct antiviral effect on herpes simplex virus types I and II, human immunodeficiency virus (HIV), hepatitis B virus, vesicular stomatitis virus, influenza virus, Epstein-Barr virus, and others. Moreover, metabolites of basidiomycetes increased antiviral immunity. PMID:25975107

  20. The cadmium-mercaptoacetic acid complex contributes to the genotoxicity of mercaptoacetic acid-coated CdSe-core quantum dots.

    Tang, Weikun; Fan, Junpeng; He, Yide; Huang, Bihai; Liu, Huihui; Pang, Daiwen; Xie, Zhixiong

    2012-01-01

    Quantum dots (QDs) have many potential clinical and biological applications because of their advantages over traditional fluorescent dyes. However, the genotoxicity potential of QDs still remains unclear. In this paper, a plasmid-based system was designed to explore the genotoxic mechanism of QDs by detecting changes in DNA configuration and biological activities. The direct chemicobiological interactions between DNA and mercaptoacetic acid-coated CdSecore QDs (MAA-QDs) were investigated. After incubation with different concentrations of MAA-QDs (0.043, 0.13, 0.4, 1.2, and 3.6 μmol/L) in the dark, the DNA conversion of the covalently closed circular (CCC) DNA to the open circular (OC) DNA was significantly enhanced (from 13.9% ± 2.2% to 59.9% ± 12.8%) while the residual transformation activity of plasmid DNA was greatly decreased (from 80.7% ± 12.8% to 13.6% ± 0.8%), which indicated that the damages to the DNA structure and biological activities induced by MAA-QDs were concentration-dependent. The electrospray ionization mass spectrometry data suggested that the observed genotoxicity might be correlated with the cadmium-mercaptoacetic acid complex (Cd-MAA) that is formed in the solution of MAA-QDs. Circular dichroism spectroscopy and transformation assay results indicated that the Cd-MAA complex might interact with DNA through the groove-binding mode and prefer binding to DNA fragments with high adenine and thymine content. Furthermore, the plasmid transformation assay could be used as an effective method to evaluate the genotoxicities of nanoparticles. PMID:22679373

  1. The cadmium–mercaptoacetic acid complex contributes to the genotoxicity of mercaptoacetic acid-coated CdSe-core quantum dots

    Tang, Weikun; Fan, Junpeng; He, Yide; Huang, Bihai; Liu, Huihui; Pang, Daiwen; Xie, Zhixiong

    2012-01-01

    Quantum dots (QDs) have many potential clinical and biological applications because of their advantages over traditional fluorescent dyes. However, the genotoxicity potential of QDs still remains unclear. In this paper, a plasmid-based system was designed to explore the genotoxic mechanism of QDs by detecting changes in DNA configuration and biological activities. The direct chemicobiological interactions between DNA and mercaptoacetic acid-coated CdSecore QDs (MAA–QDs) were investigated. After incubation with different concentrations of MAA–QDs (0.043, 0.13, 0.4, 1.2, and 3.6 μmol/L) in the dark, the DNA conversion of the covalently closed circular (CCC) DNA to the open circular (OC) DNA was significantly enhanced (from 13.9% ± 2.2% to 59.9% ± 12.8%) while the residual transformation activity of plasmid DNA was greatly decreased (from 80.7% ± 12.8% to 13.6% ± 0.8%), which indicated that the damages to the DNA structure and biological activities induced by MAA–QDs were concentration-dependent. The electrospray ionization mass spectrometry data suggested that the observed genotoxicity might be correlated with the cadmium–mercaptoacetic acid complex (Cd–MAA) that is formed in the solution of MAA–QDs. Circular dichroism spectroscopy and transformation assay results indicated that the Cd–MAA complex might interact with DNA through the groove-binding mode and prefer binding to DNA fragments with high adenine and thymine content. Furthermore, the plasmid transformation assay could be used as an effective method to evaluate the genotoxicities of nanoparticles. PMID:22679373

  2. The metabolite profiling of coastal coccolithophorid species Pleurochrysis carterae (Haptophyta)

    Zhou, Chengxu; Luo, Jie; Ye, Yangfang; Yan, Xiaojun; Liu, Baoning; Wen, Xin

    2015-11-01

    Pleurochrysis carterae is a calcified coccolithophorid species that usually blooms in the coastal area and causes aquaculture losses. The cellular calcification, blooming and many other critical species specific eco-physiological processes are closely related to various metabolic pathways. The purpose of this study is to apply the unbiased and non-destructive method of nuclear magnetic resonance (NMR) to detect the unknown holistic metabolite of P. carterae. The results show that NMR spectroscopic method is practical in the analysis of metabolites of phytoplankton. The metabolome of P. carterae was dominated by 26 metabolites involved in a number of different primary and secondary metabolic pathways. Organic acids and their derivatives, amino acids, sugars, nucleic aides were mainly detected. The abundant metabolites are that closely related to the process of cellular osmotic adjustment, which possibly reflect the active ability of P. carterae to adapt to the versatile coastal niche. DMSP (dimethylsulphoniopropionate) was the most dominant metabolite in P. carterae, up to 2.065±0.278 mg/g lyophilized cells, followed by glutamate and lactose, the contents were 0.349±0.035 and 0.301±0.073 mg/g lyophilized cells respectively. Other metabolites that had the content ranged between 0.1-0.2 mg/g lyophilized cells were alanine, isethionate and arabinose. Amino acid (valine, phenylalanine, isoleucine, tyrosine), organic acid salts (lactate, succinate), scyllitol and uracil had content ranged from 0.01 to below 0.1 mg/g lyophilized cells. Trigonelline, fumarate and formate were detected in very low content (only thousandths of 1 mg per gram of lyophilized cells or below). Our results of the holistic metabolites of P. carterae are the basic references for the further studies when multiple problems will be addressed to this notorious blooming calcifying species.

  3. The metabolite profiling of coastal coccolithophorid species Pleurochrysis carterae (Haptophyta)

    Zhou, Chengxu; Luo, Jie; Ye, Yangfang; Yan, Xiaojun; Liu, Baoning; Wen, Xin

    2016-07-01

    Pleurochrysis carterae is a calcified coccolithophorid species that usually blooms in the coastal area and causes aquaculture losses. The cellular calcification, blooming and many other critical species specific eco-physiological processes are closely related to various metabolic pathways. The purpose of this study is to apply the unbiased and non-destructive method of nuclear magnetic resonance (NMR) to detect the unknown holistic metabolite of P. carterae. The results show that NMR spectroscopic method is practical in the analysis of metabolites of phytoplankton. The metabolome of P. carterae was dominated by 26 metabolites involved in a number of different primary and secondary metabolic pathways. Organic acids and their derivatives, amino acids, sugars, nucleic aides were mainly detected. The abundant metabolites are that closely related to the process of cellular osmotic adjustment, which possibly reflect the active ability of P. carterae to adapt to the versatile coastal niche. DMSP (dimethylsulphoniopropionate) was the most dominant metabolite in P. carterae, up to 2.065±0.278 mg/g lyophilized cells, followed by glutamate and lactose, the contents were 0.349±0.035 and 0.301±0.073 mg/g lyophilized cells respectively. Other metabolites that had the content ranged between 0.1-0.2 mg/g lyophilized cells were alanine, isethionate and arabinose. Amino acid (valine, phenylalanine, isoleucine, tyrosine), organic acid salts (lactate, succinate), scyllitol and uracil had content ranged from 0.01 to below 0.1 mg/g lyophilized cells. Trigonelline, fumarate and formate were detected in very low content (only thousandths of 1 mg per gram of lyophilized cells or below). Our results of the holistic metabolites of P. carterae are the basic references for the further studies when multiple problems will be addressed to this notorious blooming calcifying species.

  4. Secondary Metabolites from Higher Fungi: Discovery, Bioactivity, and Bioproduction

    Zhong, Jian-Jiang; Xiao, Jian-Hui

    Medicinal higher fungi such as Cordyceps sinensis and Ganoderma lucidum have been used as an alternative medicine remedy to promote health and longevity for people in China and other regions of the world since ancient times. Nowadays there is an increasing public interest in the secondary metabolites of those higher fungi for discovering new drugs or lead compounds. Current research in drug discovery from medicinal higher fungi involves a multifaceted approach combining mycological, biochemical, pharmacological, metabolic, biosynthetic and molecular techniques. In recent years, many new secondary metabolites from higher fungi have been isolated and are more likely to provide lead compounds for new drug discovery, which may include chemopreventive agents possessing the bioactivity of immunomodulatory, anticancer, etc. However, numerous challenges of secondary metabolites from higher fungi are encountered including bioseparation, identification, biosynthetic metabolism, and screening model issues, etc. Commercial production of secondary metabolites from medicinal mushrooms is still limited mainly due to less information about secondary metabolism and its regulation. Strategies for enhancing secondary metabolite production by medicinal mushroom fermentation include two-stage cultivation combining liquid fermentation and static culture, two-stage dissolved oxygen control, etc. Purification of bioactive secondary metabolites, such as ganoderic acids from G. lucidum, is also very important to pharmacological study and future pharmaceutical application. This review outlines typical examples of the discovery, bioactivity, and bioproduction of secondary metabolites of higher fungi origin.

  5. Accumulation in murine amniotic fluid of halothane and its metabolites.

    Danielsson, B R; Ghantous, H; Dencker, L

    1984-11-01

    The distribution of radioactivity in pregnant mice was registered at 0, 4, and 24 hrs after a 10 min. period of inhalation of 14C-halothane. Autoradiographic methods were used to allow to distinguish between the distribution of volatile (non-metabolize) halothane, water-soluble metabolites, and firmly tissue-bound metabolites. While volatile radioactivity was seen predominantly at short survival intervals, e.g. in body fat, blood, brain and liver, metabolites accumulated with time. Peak values occurred at 4 hrs in most organs (measured with liquid scintillation as well). The most remarkable findings were the high concentrations of radioactivity in amniotic fluid (and the ocular fluids of adults) with peak values at 4 hrs and rather high concentrations still prevailing at 24 hrs after inhalation. It is assumed that this activity represents only partly volaile halothane and mostly non-volatile metabolites. High activity of metabolites was seen in the neuroepithelium of the embryo in early gestation. Firmly tissue-bound metabolites, still remaining after washing the tissues with trichloroacetic acid and organic solvents, were found in the nasal mucosa, trachea and bronchial tree and in (presumably centrilobular) zones of the liver of adults after inhalation and 5-day old mice after intraperitoneal injection, indicating the formation of reactive metabolites in these organs. Firmly tissue-bound activity was not observed in the corresponding foetal organs. PMID:6528811

  6. Studies on size variation of U(IV) colloids formed in aqueous nitric acid solutions. Contributed Paper RD-06

    Tetravalent Uranium readily undergoes hydrolysis even in highly acidic aqueous solutions. In the present work, solutions with concentration and pH ranges of 0.4 - 19 mM (total U) and 1- 4 respectively were investigated by light scattering technique with special emphasis on polymerization leading to colloid formation. Size variation of colloids formed at different pH was monitored. The results clearly indicate that the concentration has significant effect on particle size as well as stability of colloids. With increasing concentration, the size of colloids formed is smaller due to more crystalline nature of the colloids. Stability of colloids formed at lower concentration is greater than that of colloids formed at higher concentration. The results of this work are a clear indication that U(IV) hydrolysis does not differ from that of Pu(IV). (author)

  7. Contribution of carbon fixed by Rubisco and PEPC to phloem export in the Crassulacean acid metabolism plant Kalanchoe daigremontiana.

    Wild, Birgit; Wanek, Wolfgang; Postl, Wolfgang; Richter, Andreas

    2010-03-01

    Crassulacean acid metabolism (CAM) plants exhibit a complex interplay between CO(2) fixation by phosphoenolpyruvate carboxylase (PEPC) and ribulose-1,5-bisphosphate carboxylase oxygenase (Rubisco), and carbon demand for CAM maintenance and growth. This study investigated the flux of carbon from PEPC and direct Rubisco fixation to different leaf carbon pools and to phloem sap over the diurnal cycle. Concentrations and carbon isotope compositions of starch, soluble sugars, and organic acids were determined in leaves and phloem exudates of Kalanchoë daigremontiana Hamet et Perr., and related to CO(2) fixation by PEPC and Rubisco. Three types of leaf carbon pools could be distinguished. (i) Starch and malate pools were dominant and showed a pattern of reciprocal mobilization and accumulation (85/54 and 13/48 mg C g(-1) DW, respective, at the beginning/end of phase I). The carbon isotope composition of these pools was compatible with predominant PEPC fixation (delta(13)C values of -13 and -11 per thousand for starch and malate compared to -11 per thousand of PEPC fixed carbon). (ii) Isotopic composition (-17 per thousand and -14 per thousand) and concentration of glucose and fructose (2 and 3 mg C g(-1) DW, respectively) were not affected by diurnal metabolism, suggesting a low turnover. (iii) Sucrose (1-3 mg C g(-1) DW), in contrast, exhibited large diurnal changes in delta(13)C values (from -17 per thousand in the evening to -12 per thousand in the morning), which were not matched by net changes in sucrose concentration. This suggests a high sucrose turnover, fed by nocturnal starch degradation and direct Rubisco fixation during the day. A detailed dissection of the carbon fixation and mobilization pattern in K. daigremontiana revealed that direct fixation of Rubisco during the light accounted for 30% of phloem sucrose, but only 15% of fixed carbon, indicating that carbon from direct Rubisco fixation was preferentially used for leaf export. PMID:20159885

  8. The Contribution of the Activation Entropy to the Gas-Phase Stability of Modified Nucleic Acid Duplexes

    Hari, Yvonne; Dugovič, Branislav; Istrate, Alena; Fignolé, Annabel; Leumann, Christian J.; Schürch, Stefan

    2016-07-01

    Tricyclo-DNA (tcDNA) is a sugar-modified analogue of DNA currently tested for the treatment of Duchenne muscular dystrophy in an antisense approach. Tandem mass spectrometry plays a key role in modern medical diagnostics and has become a widespread technique for the structure elucidation and quantification of antisense oligonucleotides. Herein, mechanistic aspects of the fragmentation of tcDNA are discussed, which lay the basis for reliable sequencing and quantification of the antisense oligonucleotide. Excellent selectivity of tcDNA for complementary RNA is demonstrated in direct competition experiments. Moreover, the kinetic stability and fragmentation pattern of matched and mismatched tcDNA heteroduplexes were investigated and compared with non-modified DNA and RNA duplexes. Although the separation of the constituting strands is the entropy-favored fragmentation pathway of all nucleic acid duplexes, it was found to be only a minor pathway of tcDNA duplexes. The modified hybrid duplexes preferentially undergo neutral base loss and backbone cleavage. This difference is due to the low activation entropy for the strand dissociation of modified duplexes that arises from the conformational constraint of the tc-sugar-moiety. The low activation entropy results in a relatively high free activation enthalpy for the dissociation comparable to the free activation enthalpy of the alternative reaction pathway, the release of a nucleobase. The gas-phase behavior of tcDNA duplexes illustrates the impact of the activation entropy on the fragmentation kinetics and suggests that tandem mass spectrometric experiments are not suited to determine the relative stability of different types of nucleic acid duplexes.

  9. The Contribution of the Activation Entropy to the Gas-Phase Stability of Modified Nucleic Acid Duplexes

    Hari, Yvonne; Dugovič, Branislav; Istrate, Alena; Fignolé, Annabel; Leumann, Christian J.; Schürch, Stefan

    2016-04-01

    Tricyclo-DNA (tcDNA) is a sugar-modified analogue of DNA currently tested for the treatment of Duchenne muscular dystrophy in an antisense approach. Tandem mass spectrometry plays a key role in modern medical diagnostics and has become a widespread technique for the structure elucidation and quantification of antisense oligonucleotides. Herein, mechanistic aspects of the fragmentation of tcDNA are discussed, which lay the basis for reliable sequencing and quantification of the antisense oligonucleotide. Excellent selectivity of tcDNA for complementary RNA is demonstrated in direct competition experiments. Moreover, the kinetic stability and fragmentation pattern of matched and mismatched tcDNA heteroduplexes were investigated and compared with non-modified DNA and RNA duplexes. Although the separation of the constituting strands is the entropy-favored fragmentation pathway of all nucleic acid duplexes, it was found to be only a minor pathway of tcDNA duplexes. The modified hybrid duplexes preferentially undergo neutral base loss and backbone cleavage. This difference is due to the low activation entropy for the strand dissociation of modified duplexes that arises from the conformational constraint of the tc-sugar-moiety. The low activation entropy results in a relatively high free activation enthalpy for the dissociation comparable to the free activation enthalpy of the alternative reaction pathway, the release of a nucleobase. The gas-phase behavior of tcDNA duplexes illustrates the impact of the activation entropy on the fragmentation kinetics and suggests that tandem mass spectrometric experiments are not suited to determine the relative stability of different types of nucleic acid duplexes.

  10. The cadmium–mercaptoacetic acid complex contributes to the genotoxicity of mercaptoacetic acid-coated CdSe-core quantum dots

    Tang WK

    2012-05-01

    Full Text Available Weikun Tang,1 Junpeng Fan,1 Yide He,1 Bihai Huang,2 Huihui Liu,1 Daiwen Pang,2 Zhixiong Xie11College of Life Sciences, 2College of Chemistry and Molecular Sciences, Wuhan University, Wuhan, People's Republic of ChinaAbstract: Quantum dots (QDs have many potential clinical and biological applications because of their advantages over traditional fluorescent dyes. However, the genotoxicity potential of QDs still remains unclear. In this paper, a plasmid-based system was designed to explore the genotoxic mechanism of QDs by detecting changes in DNA configuration and biological activities. The direct chemicobiological interactions between DNA and mercaptoacetic acid-coated CdSe-core QDs (MAA–QDs were investigated. After incubation with different concentrations of MAA–QDs (0.043, 0.13, 0.4, 1.2, and 3.6 µmol/L in the dark, the DNA conversion of the covalently closed circular (CCC DNA to the open circular (OC DNA was significantly enhanced (from 13.9% ± 2.2% to 59.9% ± 12.8% while the residual transformation activity of plasmid DNA was greatly decreased (from 80.7% ± 12.8% to 13.6% ± 0.8%, which indicated that the damages to the DNA structure and biological activities induced by MAA–QDs were concentration-dependent. The electrospray ionization mass spectrometry data suggested that the observed genotoxicity might be correlated with the cadmium–mercaptoacetic acid complex (Cd–MAA that is formed in the solution of MAA–QDs. Circular dichroism spectroscopy and transformation assay results indicated that the Cd–MAA complex might interact with DNA through the groove-binding mode and prefer binding to DNA fragments with high adenine and thymine content. Furthermore, the plasmid transformation assay could be used as an effective method to evaluate the genotoxicities of nanoparticles.Keywords: genotoxicity, MAA CdSe quantum dots, cadmium–MAA complex, transformation assay, DNA 

  11. Exposure to benzene metabolites causes oxidative damage in Saccharomyces cerevisiae.

    Raj, Abhishek; Nachiappan, Vasanthi

    2016-06-01

    Hydroquinone (HQ) and benzoquinone (BQ) are known benzene metabolites that form reactive intermediates such as reactive oxygen species (ROS). This study attempts to understand the effect of benzene metabolites (HQ and BQ) on the antioxidant status, cell morphology, ROS levels and lipid alterations in the yeast Saccharomyces cerevisiae. There was a reduction in the growth pattern of wild-type cells exposed to HQ/BQ. Exposure of yeast cells to benzene metabolites increased the activity of the anti-oxidant enzymes catalase, superoxide dismutase and glutathione peroxidase but lead to a decrease in ascorbic acid and reduced glutathione. Increased triglyceride level and decreased phospholipid levels were observed with exposure to HQ and BQ. These results suggest that the enzymatic antioxidants were increased and are involved in the protection against macromolecular damage during oxidative stress; presumptively, these enzymes are essential for scavenging the pro-oxidant effects of benzene metabolites. PMID:27016252

  12. Spatio-temporal distribution and natural variation of metabolites in citrus fruits.

    Wang, Shouchuang; Tu, Hong; Wan, Jian; Chen, Wei; Liu, Xianqing; Luo, Jie; Xu, Juan; Zhang, Hongyan

    2016-05-15

    To study the natural variation and spatio-temporal accumulation of citrus metabolites, liquid chromatography tandem mass spectrometry (LC-MS) based metabolome analysis was performed on four fruit tissues (flavedo, albedo, segment membrane and juice sacs) and different Citrus species (lemon, pummelo and grapefruit, sweet orange and mandarin). Using a non-targeted metabolomics approach, more than 2000 metabolite signals were detected, from which more than 54 metabolites, including amino acids, flavonoids and limonoids, were identified/annotated. Differential accumulation patterns of both primary metabolites and secondary metabolites in various tissues and species were revealed by our study. Further investigation indicated that flavedo accumulates more flavonoids while juice sacs contain more amino acids. Besides this, cluster analysis based on the levels of metabolites detected in 47 individual Citrus accessions clearly grouped them into four distinct clusters: pummelos and grapefruits, lemons, sweet oranges and mandarins, while the cluster of pummelos and grapefruits lay distinctly apart from the other three species. PMID:26775938

  13. 乳酸菌及其代谢产物对刺参幼体肠道菌群和非特异性免疫的影响%The influence of lactic acid bacteria and metabolites on intes-tinal microflora and nonspecific immunity of juvenile sea cu-cumber (Apostichopus japonicus)

    宫魁; 王宝杰; 刘梅; 蒋克勇; 邱楚雯; 骆作勇; 范瑞用; 王雷

    2013-01-01

    A 60-day feeding experiment was conducted to evaluate the effects of lactic acid bacteria and its me-tabolites on bacteria of intestine, nonspecific immunity and growth of juvenile sea cucumber with initial body weight of (2.74 ± 0.17) g. Lactic acid bacteria and metabolites were added in the basal diet with 2 mL/m3 (translated by farming water). Compared with control group, the lactic acid bacteria and metabolites added to the feed signifi-cantly reduced the number of intestinal Vibrio and E. coli (P<0.05). The diet containing lactic acid bacteria and metabolites can increase the activities of acid phosphatase, alkaline phosphatase, and lysozyme significantly. There was no significant difference in activities of immune enzyme between the experimental groups. The results showed that the specific growth rate (RSG) of sea cucumber in the experimental groups were significantly higher (P<0.05) than that of the control group. These results indicated that both lactic acid bacteria and metabolites play an impor-tant role on intestinal microflora balance, nonspecific immunity and growth of sea cucumber, but the difference between the experimental groups was not significant. The metabolites may play a major role of probiotics in the process of sea cucumber farming.%以初始体质量为(2.74±0.17)g的刺参(Apostichopus japonicus)为研究对象,在室内养殖池进行60 d饲喂实验,研究饲料中添加乳酸菌制剂和乳酸菌代谢产物对刺参肠道菌群、部分免疫指标及生长的影响。在基础饲料中分别添加乳酸菌制剂和乳酸菌代谢产物各2 mL/m3(按饲养水体折算),配制2种实验饲料。实验结果显示:与对照组比,饲料中添加乳酸菌与乳酸菌代谢产物均显著降低了刺参肠道弧菌数及大肠杆菌数(P<0.05);饲料中添加乳酸菌及其代谢产物能使刺参肠道酸性磷酸酶和碱性磷酸酶活力显著升高,溶菌酶活性有所提升,其中乳酸菌制剂实验组与代

  14. Identification of Intermediate and Branch Metabolites Resulting from Biotransformation of 2-Benzoxazolinone by Fusarium verticillioides

    Glenn, A. E.; Meredith, F I; Morrison III, W. H.; Bacon, C W

    2003-01-01

    Detoxification of the maize (Zea mays) antimicrobial compound 2-benzoxazolinone by the fungal endophyte Fusarium verticillioides involves two genetic loci, FDB1 and FDB2, and results in the formation of N-(2-hydroxyphenyl)malonamic acid. Intermediate and branch metabolites were previously suggested to be part of the biotransformation pathway. Evidence is presented here in support of 2-aminophenol as the intermediate metabolite and 2-acetamidophenol as the branch metabolite, which was previous...

  15. Analysis of the contribution of acid phosphatase to P efficiency in Brassica napus under low phosphorus conditions

    2010-01-01

    To understand whether genotypic variation in acid phosphatase (APase) activity in rapeseed (Brassica napus L.) induced by phosphorus (P) deficiency has impact on P efficiency,soil APase activity in the rhizosphere for rapeseed P-efficient genotype 102 and P-inefficient genotype 105 was measured against organic and inorganic P sources in the pot experiment,and the activities of root-secreted APase and leaf intracellular APase were investigated in different P-starvation periods in the nutrient solution.Higher activity of root-secreted APase in B.napus was induced under low P conditions.However,P nutrition and P uptake efficiency of the plants supplied with organic P were not directly related to the activity of root-secreted APase due to several confounding factors affecting APase availability.The higher activity of leaf APase improved P remobilization in plants and played important roles in enhancing P use efficiency,shown by the significant correlation between leaf APase activity and P use efficiency in a rapeseed recombinant inbred population of 135 lines.

  16. Food sources of arachidonic acid (PFA 20:4), listed in descending order by percentages of their contribution to intake, based on data from the National Health and Nutrition Examination Survey 2005-2006

    Food sources of arachidonic acid (PFA 20:4), listed in descending order by percentages of their contribution to intake, based on data from the National Health and Nutrition Examination Survey 2005-2006

  17. Food sources of alpha-linolenic acid (PFA 18:3), listed in descending order by percentages of their contribution to intake, based on data from the National Health and Nutrition Examination Survey 2005-2006

    Food sources of alpha-linolenic acid (PFA 18:3), listed in descending order by percentages of their contribution to intake, based on data from the National Health and Nutrition Examination Survey 2005-2006

  18. Food sources of total omega 6 fatty acids (18:2 + 20:4), listed in descending order by percentages of their contribution to intake, based on data from the National Health and Nutrition Examination Survey 2005-2006

    Food sources of total omega 6 fatty acids (18:2 + 20:4), listed in descending order by percentages of their contribution to intake, based on data from the National Health and Nutrition Examination Survey 2005-2006

  19. [Study on secondary metabolites of endophytic fungi Penicillium dangeardii].

    Lv, Hai-ning; Ding, Guang-zhi; Liu, Yun-bao; Qu, Jing

    2015-05-01

    Endophytic fungi Penicillium dangeardii, isolated from Lysidice rhodostegia Hance root, was fermented and the secondary metabolites were studied. By means of Sephadex LH-20 column chromatography, ODS column chromatography and PHPLC over the fermented culture, 5 compounds were isolated. By using ESI-MS and NMR, the structures of the compounds were determined as N-[9-(β- D-ribofuranosyl)-9H-purin-6-yl]-L-aspartic acid (1), 3-caffeoylquinic acid (2), 4-caffeoylquinic acid (3), and 5-caffeoylquinic acid (4), 3-hydroxy-benzoic acid-4-O-β-D-glucopyranoside (5). PMID:26323144

  20. Sun, shade, and secondary metabolites

    My research program focuses on understanding plant primary and secondary metabolites. Grape secondary metabolites, such as phenolics, have long been valuable for the organoleptic properties they impart to fruit and wine, and, more recently, for their possible health benefits. These compounds develop...

  1. Secondary metabolites from Eremostachys laciniata

    Calis, Ihsan; Güvenc, Aysegül; Armagan, Metin;

    2008-01-01

    metabolites were elucidated from spectroscopic (UV, IR, 1D- and 2D-NMR) and ESI-MS evidence, as well as from their specific optical rotation. The presence of these metabolites of three different classes strongly supports the close relationship of the genera Eremostachys and Phlomis....

  2. Characterization of rhizobacteria associated to maize crop in IAA, siderophores and salicylic acid metabolite production Caracterización de rizobacterias asociadas al cultivo del maíz en la producción de metabolitos del tipo AIA, sideróforos y ácido salicílico

    Heydrich Mayra

    2004-07-01

    Full Text Available It has been demonstrated that rhizobacteria are able to produce metabolites having agricultural interest, including salicylic acid, the siderophores and phytohormones. Indol acetic acid (IAA is the most well-known and studied auxin, playing a governing role in culture growth. The object of this work was to characterise rhizobacteria associated with the maize crop in terms of producing IAA, siderophores and salicylic acid metabolites. Burkholderia cepacia and Pseudomonas fluorescens strains previously isolated from maize Francisco variety rhizosphere were used. Colorimetric and chromatographic techniques for detecting these metabolites were studied; multi-variable analysis of hierarchic conglomerate and complete ligament were used for selecting the best strains for producing metabolites of interest. These results demonstrated that all rhizobacteria strains studied produced IAA, siderophores and salicylic acid metabolites. Burkholderia cepacia MBf21, MBp1, MBp2, MBf22, MBp3, MBf20, MBf 15 and Pseudomonas fluorescens MPp4strains have presented the greatest production of these metabolites, showing that these strains could be used in promoting vegetal growth in economically important cultures. Key words: Pseudomonas fluorescens, Burkholderia cepacia, IAA, siderophore, salicylic acid.Se ha demostrado que las rizobacterias tienen la capacidad de producir metabolitos de interés agrícola, entre los que se destacan el ácido salicílico, los sideróforos y las f itohormonas; dentro de estas últimas, el ácido Indol acético (AIA es la auxina más conocida y estudiada, la cual desempeña un papel rector en el crecimiento de los cultivos. Esta investigación se realizó con el objetivo de caracterizar rizobacterias asociadas al cultivo del maíz en cuanto a la producción de AIA, sideróforos y ácido salicílico. Para ello se empleó un conjunto de cepas de Burkholderia cepacia y Pseudomonas fluorescens previamente aisladas de la rizosfera del ma

  3. LC-MS/MS Method for the Determination and Quantitation of Penicillin G and Its Metabolites in Citrus Fruits Affected by Huanglongbing.

    Aldeek, Fadi; Rosana, Michael R; Hamilton, Zaid K; Crosswhite, Mark R; Burrows, Casey W; Singh, Sonal; Gerard, Ghislain; Hammack, Walter; Cook, Jo-Marie

    2015-07-01

    In this study, we developed and validated a method for the extraction, identification, and quantitation of penicillin G and its metabolites (penilloic acid and penillic acid) in a variety of citrus fruits by employing sequential liquid/liquid and solid-phase extraction techniques in conjunction with UHPLC-MS/MS. Two product ion transitions per analyte were required for identification, which contributes to a high degree of selectivity. Corrected recoveries of penicillin G using an isotopically labeled internal standard were 90-100% at fortification levels of 0.1, 0.25, 1, and 10 ng/g. Absolute recoveries for penillic acid and penilloic acid were 50-75% depending on the matrix used. The limit of detection (LOD) of penicillin G and its metabolites was found to be 0.1 ng/g when 2 g of citrus was extracted. This method is useful in determining residue levels of penicillin G and its metabolites in citrus trees infected with huanglongbing bacteria after antibiotic treatment. PMID:26072945

  4. EFSA Panel on Dietetic Products, Nutrition and Allergies (NDA); Scientific Opinion on the substantiation of health claims related to docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA) and gamma-linolenic acid (GLA) and contribution to normal cognitive function (ID 532) and maintenance of

    Tetens, Inge

    claims in relation to docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA) and gamma-linolenic acid (GLA) and contribution to normal cognitive function and maintenance of normal bone. The scientific substantiation is based on the information provided by the Member States in the consolidated list of...... (EPA) in fish oil. The Panel considers that the food constituents, DHA, EPA and GLA, are sufficiently characterised....... Article 13 health claims and references that EFSA has received from Member States or directly from stakeholders. The food constituents that are the subjects of the health claims are “omega-3 and omega-6 fatty acids (GLA)”, “gamma-linolenic acid + eicosapentaenoic acid (GLA+EPA)”, and “evening primrose oil...

  5. A selective metabolite array for the detection of phosphometabolites

    Highlights: ► Metal affinity based selective array for the screening of phosphometabolites. ► A simple one-step plasma polymer coating and direct metal immobilisation. ► ToF-SIMS detection of phosphometabolite array. ► Ga and Zr suitable metals. - Abstract: Immobilised metal ion affinity (IMA) has been traditionally used specifically for the separation of phosphorylated proteins and nucleic acids, in proteomics and genomics, respectively. This report describes the novel application of IMA in metabolomics for the development of metabolite arrays to detect phosphometabolites using a plasma polymer-modified surface. Immobilisation of gallium, zirconium, cobalt, copper, zinc, nickel, iron, and chromium to acrylic acid plasma polymer followed by subsequent exposure to metabolites (phospho- and non-phosphometabolites) was investigated. Results analysed using ToF-SIMS suggests that gallium and zirconium exhibit higher phosphometabolite affinity and specificity compared to other metals, and can be used to develop metabolite arrays for the detection of phosphometabolites.

  6. Contribution of Root Traits to Phosphorus Acqusition Efficiency by Maize Landraces in Acid Soils of the Highlands in Central Mexico

    evidenced by increased plant biomass and grain yield. Late maturity accessions had improved growth and yield under low P. Accessions DP x Tromba, HV313 x DEM, Macho III-04, and CIMMYT-1, were categorized as the most P efficient under low P and as the most responsive to increased P availability on a P-deficient acidic soil of this region. P-efficient accessions such as DP x Tromba accessed soil P not available to P-inefficient accessions. These results indicate that landraces of the Central Mexican highlands exhibit variation for several root traits that may be useful for genetic improvement of P acquisition efficiency in maize. (author)

  7. Arachidonic acid metabolism by bovine placental tissue during the last month of pregnancy

    Conversion of tritiated arachidonic acid (AA) into metabolites of the cyclo- and lipoxygenase pathways by bovine fetal placental tissue (200 mg) and fetal plus maternal placental tissue (400 mg) of Days 255, 265, 275 of gestation and at parturition (n = 5) during a 30 min incubation was measured using reverse-phase high pressure liquid chromatography. Fetal placental tissue produced 13,14-dihydro-15-keto-prostaglandin E2 (PGEM) as the major metabolite, the synthesis of which increased from Day 265 to Day 275 and parturition by 150% and 475%, respectively. In tissues collected at parturition, PGE2 synthesis was also detected. On Day 275 and at parturition fetal placental tissue synthesized the metabolite 12-hydroxyheptadecatrienoic acid (HHT), and throughout the experimental period the lipoxygenase product 15-HETE was detected with synthesis rates increasing over time of gestation. In addition, an unidentified metabolite was regularly found in the radiochromatograms which eluted at 1 h and 1 min (U101), between HHT and 15-HETE. The synthesis of this metabolite decreased as pregnancy progressed. Furthermore, various other polar and nonpolar metabolites pooled under the heading UNID were eluted, the production of which increased over time of gestation. The presence of maternal placental tissue did not influence the synthesis of PGEM, 15-HETE and U101, but the production of HHT was decreased when maternal tissue was present. Also, as pregnancy progressed, maternal placental tissue seemed to contribute to the pool of unidentified metabolites. In conclusion, fetal placental tissue seems to be the major source of the AA metabolites when compared with maternal placental tissue, and AA metabolism by bovine placental tissue is markedly increased throughout the last month of pregnancy, suggesting a role for AA metabolites in mechanisms controlling parturition

  8. Salicylic acid: a link between aspirin, diet and the prevention of colorectal cancer.

    Paterson, J R; Lawrence, J R

    2001-08-01

    Aspirin was introduced into clinical practice more than 100 years ago. This unique drug belongs to a family of compounds called the salicylates, the simplest of which is salicylic acid, the principal metabolite of aspirin. Salicylic acid is responsible for the anti-inflammatory action of aspirin, and may cause the reduced risk of colorectal cancer observed in those who take aspirin. Yet salicylic acid and other salicylates occur naturally in fruits and plants, while diets rich in these are believed to reduce the risk of colorectal cancer. Serum salicylic acid concentrations are greater in vegetarians than non-vegetarians, and there is overlap between concentrations in vegetarians and those taking low-dose aspirin. We propose that the cancer-preventive action of aspirin is due to its principal metabolite, salicylic acid, and that dietary salicylates can have the same effect. It is also possible that natural salicylates contribute to the other recognized benefits of a healthy diet. PMID:11493722

  9. Effects of Kynurenine Pathway Metabolites on Intracellular NAD+ Synthesis and Cell Death in Human Primary Astrocytes and Neurons

    Nady Braidy

    2009-01-01

    Full Text Available The kynurenine pathway (KP is a major route of L-tryptophan catabolism resulting in the production of the essential pyridine nucleotide nicotinamide adenine dinucleotide, (NAD+. Up-regulation of the KP during inflammation leads to the release of a number of biologically active metabolites into the brain. We hypothesised that while some of the extracellular KP metabolites may be beneficial for intracellular NAD+ synthesis and cell survival at physiological concentrations, they may contribute to neuronal and astroglial dysfunction and cell death at pathophysiological concentrations. In this study, we found that treatment of human primary neurons and astrocytes with 3-hydroxyanthranilic acid (3-HAA, 3-hydroxykynurenine (3-HK, quinolinic acid (QUIN, and picolinic acid (PIC at concentrations below 100 nM significantly increased intracellular NAD+ levels compared to non-treated cells. However, a dose dependent decrease in intracellular NAD+ levels and increased extracellular LDH activity was observed in human astrocytes and neurons treated with 3-HAA, 3-HK, QUIN and PIC at concentrations 100 nM and kynurenine (KYN, at concentrations above 1 μM. Intracellular NAD+ levels were unchanged in the presence of the neuroprotectant, kynurenic acid (KYNA, and a dose dependent increase in intracellular NAD+ levels was observed for TRP up to 1 mM. While anthranilic acid (AA increased intracellular NAD+ levels at concentration below 10 μM in astrocytes. NAD+ depletion and cell death was observed in AA treated neurons at concentrations above 500 nM. Therefore, the differing responses of astrocytes and neurons to an increase in KP metabolites should be considered when assessing KP toxicity during neuroinflammation.

  10. The contribution of Fe(III) and humic acid reduction to ecosystem respiration in drained thaw lake basins of the Arctic Coastal Plain

    Lipson, David A.; Raab, Theodore K.; Goria, Dominic; Zlamal, Jaime

    2013-04-01

    research showed that anaerobic respiration using iron (Fe) oxides as terminal electron acceptor contributed substantially to ecosystem respiration (ER) in a drained thaw lake basin (DTLB) on the Arctic coastal plain. As DTLBs age, the surface organic layer thickens, progressively burying the Fe-rich mineral layers. We therefore hypothesized that Fe(III) availability and Fe reduction would decline with basin age. We studied four DTLBs across an age gradient, comparing seasonal changes in the oxidation state of dissolved and extractable Fe pools and the estimated contribution of Fe reduction to ER. The organic layer thickness did not strictly increase with age for these four sites, though soil Fe levels decreased with increasing organic layer thickness. However, there were surprisingly high levels of Fe minerals in organic layers, especially in the ancient basin where cryoturbation may have transported Fe upward through the profile. Net reduction of Fe oxides occurred in the latter half of the summer and contributed an estimated 40-45% to ecosystem respiration in the sites with the thickest organic layers and 61-63% in the sites with the thinnest organic layers. All sites had high concentrations of soluble Fe(II) and Fe(III), explained by the presence of siderophores, and this pool became progressively more reduced during the first half of the summer. Redox titrations with humic acid (HA) extracts and chelated Fe support our view that this pattern indicates the reduction of HA during this interval. We conclude that Fe(III) and HA reductions contribute broadly to ER in the Arctic coastal plain.

  11. Metabolomic investigation into variation of endogenous metabolites in professional athletes subject to strength-endurance training.

    Yan, Bei; A, Jiye; Wang, Guangji; Lu, Huali; Huang, Xiaoping; Liu, Yi; Zha, Weibin; Hao, Haiping; Zhang, Ying; Liu, Linsheng; Gu, Shenghua; Huang, Qing; Zheng, Yuanting; Sun, Jianguo

    2009-02-01

    Strength-endurance type of sport can lead to modification of human beings' physiological status. The present study aimed to investigate the alteration of metabolic phenotype or biochemical compositions in professional athletes induced by long-term training by means of a novel systematic tool, metabolomics. Resting venous blood samples of junior and senior male rowers were obtained before and after 1-wk and 2-wk training. Venous blood from healthy male volunteers as control was also sampled at rest. Endogenous metabolites in serum were profiled by GC/TOF-MS and multivariate statistical technique, i.e., principal component analysis (PCA), and partial least squares projection to latent structures and discriminant analysis (PLS-DA) were used to process the data. Significant metabolomic difference was observed between the professional athletes and control subjects. Long-term strength and endurance training induced distinct separation between athletes of different exercise seniority, and training stage-related trajectory of the two groups of athletes was clearly shown along with training time. However, most of these variations were not observed by common biochemical parameters, such as hemoglobin, testosterone, and creatine kinase. The identified metabolites contributing to the classification included alanine, lactate, beta-d-methylglucopyranoside, pyroglutamic acid, cysteine, glutamic acid, citric acid, free fatty acids, valine, glutamine, phenylalanine, tyrosine, and so on, which were involved in glucose metabolism, oxidative stress, energy metabolism, lipid metabolism, amino acid metabolism. These findings suggest that metabolomics is a promising and potential tool to profile serum of professional athletes, make a deep insight into physiological states, and clarify the disorders induced by strength-endurance physical exercise. PMID:19036890

  12. Genetic analysis of the metabolic pathways responsible for aroma metabolite production by Saccharomyces cerevisiae.

    Styger, Gustav; Jacobson, Dan; Prior, Bernard A; Bauer, Florian F

    2013-05-01

    During alcoholic fermentation, higher alcohols, esters, and acids are formed from amino acids via the Ehrlich pathway by yeast, but many of the genes encoding the enzymes have not yet been identified. When the BAT1/2 genes, encoding transaminases that deaminate amino acids in the first step of the Ehrlich pathway are deleted, higher metabolite formation is significantly decreased. Screening yeast strains with deletions of genes encoding decarboxylases, dehydrogenases, and reductases revealed nine genes whose absence had the most significant impact on higher alcohol production. The seven most promising genes (AAD6, BAT2, HOM2, PAD1, PRO2, SPE1, and THI3) were further investigated by constructing double- and triple-deletion mutants. All double-deletion strains showed a greater decrease in isobutanol, isoamyl alcohol, isobutyric, and isovaleric acid production than the corresponding single deletion strains with the double-deletion strains in combination with ∆bat2 and the ∆hom2-∆aad6 strain revealing the greatest impact. BAT2 is the dominant gene in these deletion strains and this suggests the initial transaminase step of the Ehrlich pathway is rate-limiting. The triple-deletion strains in combination with BAT2 (∆bat2-∆thi3-∆aad6 and ∆bat2-∆thi3-∆hom2) had the greatest impact on the end metabolite production with the exception of isoamyl alcohol and isovaleric acid. The strain deleted for two dehydrogenases and a reductase (∆hom2-∆pro2-∆aad6) had a greater effect on the levels of these two compounds. This study contributes to the elucidation of the Ehrlich pathway and its significance for aroma production by fermenting yeast cells. PMID:23111598

  13. Dietary Supplements and Sports Performance: Metabolites, Constituents, and Extracts

    Williams Melvin

    2006-01-01

    Abstract This is the fifth in a series of six articles to discuss the major classes of dietary supplements (vitamins; minerals; amino acids; herbs or botanicals; and metabolites/constituents/extracts). The major focus is on efficacy of such dietary supplements to enhance exercise or sports performance.

  14. Uptake of Seeds Secondary Metabolites by Virola surinamensis Seedlings

    Massuo Jorge Kato; Massayoshi Yoshida; Norberto Peporine Lopes; Denise Brentan da Silva; Alberto José Cavalheiro

    2012-01-01

    The major secondary metabolites and fatty acids occurring in the seeds of Virola surinamensis were monitored by GC-MS during germination and seedling development. The role as carbon source for seedling development was indicated considering that both classes of compounds were similarly consumed in the seeds and that no selective consumption of compounds could be detected.

  15. Dietary Supplements and Sports Performance: Metabolites, Constituents, and Extracts

    Williams Melvin

    2006-12-01

    Full Text Available Abstract This is the fifth in a series of six articles to discuss the major classes of dietary supplements (vitamins; minerals; amino acids; herbs or botanicals; and metabolites/constituents/extracts. The major focus is on efficacy of such dietary supplements to enhance exercise or sports performance.

  16. Intestinal health functions of colonic microbial metabolites: A review

    Havenaar, R.

    2011-01-01

    This review tries to find a scientific answer on the following two questions: (1) to what extent do we understand the specific role of colonic microbial metabolites, especially short-chain fatty acids (SCFA), in maintaining the health status and prevention of diseases of the colon and the host; (2)

  17. Effects of Controlled Atmospheres on Production of Sesquiterpenoid Stress Metabolites by White Potato Tuber

    Alves, Leo M.; Heisler, Edward G.; Kissinger, John C.; Patterson, Joseph M.; Kalan, Edwin B.

    1979-01-01

    Levels of katahdinone (solavetivone), lubimin, rishitin, and phytuberin, sesquiterpenoid stress metabolites of white potato (Solanum tuberosum), were monitored in tuber slices which were challenged with an extract of Phytophthora infestans and incubated under controlled atmospheres. A mixture of ethylene in air enhanced stress metabolite production. This enhancement was amplified by higher partial pressures of oxygen. Stress metabolite production was inhibited by salicylhydroxamic acid. These results suggest the involvement of cyanide-resistant respiration in the production of potato stress metabolites, compounds which may serve as phytoalexins. PMID:16660728

  18. Phenolic metabolites of anthocyanins modulate mechanisms of endothelial function

    Edwards, Michael; Czank, Charles; Woodward, Gary; Cassidy, Aedin; Kay, Colin

    2015-01-01

    Anthocyanins are reported to have vascular bioactivity, however their mechanisms of action are largely unknown. Evidence suggests that anthocyanins modulate endothelial function, potentially by increasing nitric oxide (NO) synthesis, or enhancing NO bioavailability. This study compared the activity of cyanidin-3-glucoside, its degradation product protocatechuic acid, and phase II metabolite, vanillic acid. Production of NO and superoxide and expression of endothelial NO synthase (eNOS), NADPH...

  19. Contribution of Direct and Indirect Exposure to Human Serum Concentrations of Perfluorooctanoic Acid in an Occupationally Exposed Group of Ski Waxers.

    Gomis, Melissa I; Vestergren, Robin; Nilsson, Helena; Cousins, Ian T

    2016-07-01

    The contribution of direct (i.e., uptake of perfluorooctanoic acid (PFOA) itself) and indirect (i.e., uptake of 8:2 fluorotelomer alcohol (FTOH) and metabolism to PFOA) exposure to PFOA serum concentrations was investigated using a dynamic one-compartment pharmacokinetic (PK) model. The PK model was applied to six occupationally exposed ski waxers for whom direct and indirect exposures via inhalation were characterized using multiple measurements with personal air sampling devices. The model was able to predict the diverging individual temporal trends of PFOA in serum with correlation coefficients of 0.82-0.94. For the four technicians with high initial concentrations of PFOA in serum (250-1050 ng/mL), the ongoing occupational exposure (both direct and indirect) was of minor importance and net depuration of PFOA was observed throughout the ski season. An estimated average intrinsic elimination half-life of 2.4 years (1.8-3.1 years accounting for variation between technicians and model uncertainty) was derived for these technicians. The remaining two technicians, who had much lower initial serum concentrations (10-17 ng/mL), were strongly influenced by exposure during the ski season with indirect exposure contributing to 45% of PFOA serum concentrations. On the basis of these model simulations, an average metabolism yield of 0.003 (molar concentration basis; uncertainty range of 0.0006-0.01) was derived for transformation of 8:2 FTOH to PFOA. An uncertainty analysis was performed, and it was determined that the input parameters quantifying the intake of PFOA were mainly responsible for the uncertainty of the metabolism yield and the initial concentration of PFOA in serum was mainly contributing to the uncertainty of estimated serum half-lives. PMID:27304840

  20. Effect of fertilisation and harvest period on polar metabolites of Calendula oficcinalis

    Eduardo Felipe Alves Fernandes

    2013-10-01

    Full Text Available The present study evaluated the chemical profile of polar extracts of Calendula officinalis L., Asteraceae, that were grown under different cultivation conditions: chemical fertilisation, organic fertilisation and mulching. Furthermore, we investigated metabolite variations during plant development by comparing the metabolites from harvested plants at 60 and 120 days after planting. We used HPLC-DAD-MS/MS to tentatively identify metabolites. In total, we identified seven known compounds: five flavonoid glycosides and two caffeoylquinic acids derivatives. There were no statistically significant differences in the expression of metabolites from plants grown under the examined soil treatments. However, five substances varied according to harvest time, suggesting that the biosynthesis of polar metabolites of Calendula officinalis is not affected by changes in soil composition. Therefore, this plant could represent a source for phytomedicines with a constant content of polar metabolites.

  1. 酸马奶中一株乳酸菌与一株酵母菌共生关系和风味代谢产物的研究%Interactions between lactic acid bacteria and yeasts in kumiss :research on flavour metabolites

    闫彬; 贺银凤

    2012-01-01

    对内蒙古锡盟地区酸马奶中分离出的一株乳酸菌和一株酵母菌进行混合培养,测定了双菌培养体系中的风味代谢产物,以单菌培养作为对照.结果表明:酵母菌水解蛋白能力要强于乳酸菌;双菌培养基中有利于产出更多的风味物质;乳酸菌和酵母菌在发酵过程中都可以利用苹果酸和酒石酸作为碳源进行生长,在柠檬酸的利用上,乳酸菌可以进行柠檬酸代谢,而酵母菌以产生柠檬酸为特点,暗示了二者间的相互作用关系;在双菌培养基中甲酸、乙酸、丙酸的产量在不同时期都高于单菌培养过程中的生成量,提示出乳酸菌和酵母菌存在某种共生关系.%The co- cultivation of a strain of lactic acid bacteria and a strain of yeast isolated from koumiss in Inner Mongolia ,Ximeng region was studied, flavor metabolites formed in mixed-culture medium were determined, single culture as a control, the results are as follows: the ability of hydrolyzing protein of yeast is stronger than that of lactic acid bacteria; the medium is more conducive to produce more flavor compounds when mix-cultured; lactic acid bacteria and yeast in the fermentation process can make use of malic acid and tartaric acid as carbon source for growth; on the use of citric acid, lactic acid can metabolism citric acid, while yeast characterize in producing citric acid, suggesting the interaction between the lactic acid bacteria and yeasts; the production of formic acid, acetic acid, propionic acid in co-culture medium at different times is higher than that in single-culture medium during the process of fermentation, suggesting some kind of symbiotic relationship between lactic acid bacteria and yeasts.

  2. Metabolic engineering with systems biology tools to optimize production of prokaryotic secondary metabolites.

    Kim, Hyun Uk; Charusanti, Pep; Lee, Sang Yup; Weber, Tilmann

    2016-08-27

    Covering: 2012 to 2016Metabolic engineering using systems biology tools is increasingly applied to overproduce secondary metabolites for their potential industrial production. In this Highlight, recent relevant metabolic engineering studies are analyzed with emphasis on host selection and engineering approaches for the optimal production of various prokaryotic secondary metabolites: native versus heterologous hosts (e.g., Escherichia coli) and rational versus random approaches. This comparative analysis is followed by discussions on systems biology tools deployed in optimizing the production of secondary metabolites. The potential contributions of additional systems biology tools are also discussed in the context of current challenges encountered during optimization of secondary metabolite production. PMID:27072921

  3. Metabolic engineering with systems biology tools to optimize production of prokaryotic secondary metabolites

    Kim, Hyun Uk; Charusanti, Pep; Lee, Sang Yup;

    2016-01-01

    for the optimal production of various prokaryotic secondary metabolites: native versus heterologous hosts (e.g., Escherichia coli) and rational versus random approaches. This comparative analysis is followed by discussions on systems biology tools deployed in optimizing the production of secondary metabolites......Metabolic engineering using systems biology tools is increasingly applied to overproduce secondary metabolites for their potential industrial production. In this Highlight, recent relevant metabolic engineering studies are analyzed with emphasis on host selection and engineering approaches....... The potential contributions of additional systems biology tools are also discussed in the context of current challenges encountered during optimization of secondary metabolite production....

  4. Secondary metabolites from marine microorganisms

    KELECOM ALPHONSE

    2002-01-01

    Full Text Available After 40 years of intensive research, chemistry of marine natural products has become a mature field. Since 1995, there are signals of decreased interest in the search of new metabolites from traditional sources such as macroalgae and octocorals, and the number of annual reports on marine sponges stabilized. On the contrary, metabolites from microorganisms is a rapidly growing field, due, at least in part, to the suspicion that a number of metabolites obtained from algae and invertebrates may be produced by associated microorganisms. Studies are concerned with bacteria and fungi, isolated from seawater, sediments, algae, fish and mainly from marine invertebrates such as sponges, mollusks, tunicates, coelenterates and crustaceans. Although it is still to early to define tendencies, it may be stated that the metabolites from microorganisms are in most cases quite different from those produced by the invertebrate hosts. Nitrogenated metabolites predominate over acetate derivatives, and terpenes are uncommon. Among the latter, sesquiterpenes, diterpenes and carotenes have been isolated; among nitrogenated metabolites, amides, cyclic peptides and indole alkaloids predominate.

  5. The diet-microbiota-metabolite axis regulates the host physiology.

    Yamada, Takahiro; Takahashi, Daisuke; Hase, Koji

    2016-07-01

    The intestinal microbiota has been implicated in a wide range of diseases, including inflammatory bowel disease, obesity and cancer. Food ingredients are considered a major determinant of gut microbial composition, as exemplified by high-fat diet-induced dysbiosis that can affect host physiology. Accumulating studies show that microbial metabolites are key regulators of the intestinal epithelial barrier and gut immunity. In particular, short-chain fatty acids produced by bacterial fermentation of indigestible polysaccharides have profound impacts on host physiology beyond the gut. In this review, we describe the influences of the diet-microbiota-metabolite axis on host physiology, and especially on the immune and metabolic systems. PMID:26970281

  6. An update on secondary metabolites from haloxylon species

    Secondary metabolites have been isolated from the various species of the genus Haloxylon (Chenopodiaceae) including H. ammodendron, H. aphyllum, H. articulatum, H. griffithii, H. persicum, H. recurvum, H. salicornicum, H. schmittianum Pomel and H. scoparium. These compounds belong to the classes, fatty acids and their esters, triglycerides, alcohols, steroids and their glycosides, phenol derivatives, coumarins, alkaloids of different classes, monocyclic naphthene derivatives, terpenes and flavonoids, and their glycosides. This present review will discuss the secondary metabolites 1-107 of different classes isolated from Haloxylon species with biological activities up to 2010. (author)

  7. Classes of secondary metabolites identified in three legume species

    Delci de Deus Nepomuceno

    2013-10-01

    Full Text Available The objective of this study was to identify secondary metabolite classes of tropical kudzu, perennial soybean and archer legumes through identification by phytochemical screening and confirmation by infrared (IR, ¹H and carbon-13 NMR spectroscopy of fractions obtained by organic solvents partition of crude extracts. These analyses allowed for proposing the presence of the following metabolite classes: reducing sugars, saccharides, saponins, cardiac glucosides, terpenes, coumarins, tannins, alkaloids and flavonoids. The tannins, saponins, alkaloids, organic acids and flavonoids had special consideration because they are toxic compounds and can affect the animal metabolism.

  8. Presence of toxic microbial metabolites in table olives

    Eduardo eMedina-Pradas

    2015-08-01

    Full Text Available Table olives have an enormous importance in the diet and culture of many Mediterranean countries. Albeit there are different ways to produce this fermented vegetable, brining/salting, fermentation and acidification are common practices for all of them. Preservation methods such as pasteurization or sterilization are frequently used to guarantee the stability and safety of fermented olives. However, final products are not always subjected to a heat treatment. Thus, microbiota is not always removed and appropriate levels of acidity and salt must be obtained before commercialization. Despite the physicochemical conditions not being favourable for the growth of foodborne pathogens, some illness outbreaks have been reported in the literature. Street markets, inappropriate manipulation and storage conditions were the origin of many of the samples in which foodborne pathogens or their metabolites were detected. Many authors have also studied the survival of pathogens in different styles of table olive elaboration, finding in general that olive environment is not appropriate for their presence. Inhibitory compounds such as polyphenols, low availability of nutrients, high salt content, low pH levels, bacteriocins or the addition of preservatives act as hurdles against undesirable microorganisms, which contribute to obtaining a safe and good quality product.

  9. Pattern recognition analysis of chromatographic fingerprints of Crocus sativus L. secondary metabolites towards source identification and quality control.

    Aliakbarzadeh, Ghazaleh; Sereshti, Hassan; Parastar, Hadi

    2016-05-01

    Chromatographic fingerprinting is an effective methodology for authentication and quality control of herbal products. In the presented study, a chemometric strategy based on multivariate curve resolution-alternating least squares (MCR-ALS) and multivariate pattern recognition methods was used to establish a gas chromatography-mass spectrometry (GC-MS) fingerprint of saffron. For this purpose, the volatile metabolites of 17 Iranian saffron samples, collected from different geographical regions, were determined using the combined method of ultrasound-assisted solvent extraction (UASE) and dispersive liquid-liquid microextraction (DLLME), coupled with GC-MS. The resolved elution profiles and the related mass spectra obtained by an extended MCR-ALS algorithm were then used to estimate the relative concentrations and to identify the saffron volatile metabolites, respectively. Consequently, 77 compounds with high reversed match factors (RMFs > 850) were successfully determined. The relative concentrations of these compounds were used to generate a new data set which was analyzed by multivariate data analysis methods including principal component analysis (PCA) and k-means. Accordingly, the saffron samples were categorized into five classes using these techniques. The results revealed that 11 compounds, as biomarkers of saffron, contributed to the class discrimination and characterization. Eleven biomarkers including nine secondary metabolites of saffron (safranal, α- and β-isophorone, phenylethyl alcohol, ketoisophorone, 2,2,6-trimethyl-1,4-cyclohexanedione, 2,6,6-trimethyl-4-oxo-2-cyclohexen-1-carbaldehyde, 2,4,4-trimethyl-3-carboxaldehyde-5-hydroxy-2,5-cyclohexadien-1-one, and 2,6,6-trimethyl-4-hydroxy-1-cyclohexene-1-carboxaldehyde (HTCC)), a primary metabolite (linoleic acid), and a long chain fatty alcohol (nanocosanol) were distinguished as the saffron fingerprint. Finally, the individual contribution of each biomarker to the classes was determined by the

  10. Responsiveness of Urinary and Plasma Alkylresorcinol Metabolites to Rye Intake in Finnish Women

    Mylène Aubertin-Leheudre

    2010-04-01

    Full Text Available Alkylresorcinols [ARs] have been proposed for use as biomarkers of whole-grain intake. The aim here was to examine the responsiveness of AR metabolites to rye intake. Sixty women were divided into three groups according to their rye consumption. We observed significant differences between groups in plasma 3-[3,5-dihydroxyphenyl]-1-propanoic acid [DHPPA] and in urinary DHPPA and 3,5-dihydroxybenzoic acid [DHBA]. In addition, these AR metabolites increased proportionally to rye fiber intake. We conclude that these ARs metabolites are accurate and useful biomarkers of rye fiber intake. Further studies are needed to confirm our results in larger and different populations.

  11. Alterations of urinary metabolite profile in model diabetic nephropathy

    Highlights: • 1H NMR spectroscopy was employed to study urinary metabolite profile in diabetic mouse models. • Mouse urinary metabolome showed major changes that are also found in human diabetic nephropathy. • These models can be new tools to study urinary biomarkers that are relevant to human disease. - Abstract: Countering the diabetes pandemic and consequent complications, such as nephropathy, will require better understanding of disease mechanisms and development of new diagnostic methods. Animal models can be versatile tools in studies of diabetic renal disease when model pathology is relevant to human diabetic nephropathy (DN). Diabetic models using endothelial nitric oxide synthase (eNOS) knock-out mice develop major renal lesions characteristic of human disease. However, it is unknown whether they can also reproduce changes in urinary metabolites found in human DN. We employed Type 1 and Type 2 diabetic mouse models of DN, i.e. STZ-eNOS−/− C57BLKS and eNOS−/− C57BLKS db/db, with the goal of determining changes in urinary metabolite profile using proton nuclear magnetic resonance (NMR). Six urinary metabolites with significantly lower levels in diabetic compared to control mice have been identified. Specifically, major changes were found in metabolites from tricarboxylic acid (TCA) cycle and aromatic amino acid catabolism including 3-indoxyl sulfate, cis-aconitate, 2-oxoisocaproate, N-phenyl-acetylglycine, 4-hydroxyphenyl acetate, and hippurate. Levels of 4-hydroxyphenyl acetic acid and hippuric acid showed the strongest reverse correlation to albumin-to-creatinine ratio (ACR), which is an indicator of renal damage. Importantly, similar changes in urinary hydroxyphenyl acetate and hippurate were previously reported in human renal disease. We demonstrated that STZ-eNOS−/− C57BLKS and eNOS−/− C57BLKS db/db mouse models can recapitulate changes in urinary metabolome found in human DN and therefore can be useful new tools in metabolomic

  12. The effects of Mary Rose conservation treatment on iron oxidation processes and microbial communities contributing to acid production in marine archaeological timbers.

    Joanne Preston

    Full Text Available The Tudor warship the Mary Rose has reached an important transition point in her conservation. The 19 year long process of spraying with polyethylene glycol (PEG has been completed (April 29(th 2013 and the hull is air drying under tightly controlled conditions. Acidophilic bacteria capable of oxidising iron and sulfur have been previously identified and enriched from unpreserved timbers of the Mary Rose, demonstrating that biological pathways of iron and sulfur oxidization existed potentially in this wood, before preservation with PEG. This study was designed to establish if the recycled PEG spray system was a reservoir of microorganisms capable of iron and sulfur oxidization during preservation of the Mary Rose. Microbial enrichments derived from PEG impregnated biofilm collected from underneath the Mary Rose hull, were examined to better understand the processes of cycling of iron. X-ray absorption spectroscopy was utilised to demonstrate the biological contribution to production of sulfuric acid in the wood. Using molecular microbiological techniques to examine these enrichment cultures, PEG was found to mediate a shift in the microbial community from a co-culture of Stenotrophomonas and Brevunidimonas sp, to a co-culture of Stenotrophomonas and the iron oxidising Alicyclobacillus sp. Evidence is presented that PEG is not an inert substance in relation to the redox cycling of iron. This is the first demonstration that solutions of PEG used in the conservation of the Mary Rose are promoting the oxidation of ferrous iron in acidic solutions, in which spontaneous abiotic oxidation does not occur in water. Critically, these results suggest PEG mediated redox cycling of iron between valence states in solutions of 75% PEG 200 and 50% PEG 2000 (v/v at pH 3.0, with serious implications for the future use of PEG as a conservation material of iron rich wooden archaeological artefacts.

  13. 20-Hydroxyeicosatetraenoic acid contributes to the inhibition of K+ channel activity and vasoconstrictor response to angiotensin II in rat renal microvessels.

    Fan Fan

    Full Text Available The present study examined whether 20-hydroxyeicosatetraenoic acid (HETE contributes to the vasoconstrictor effect of angiotensin II (ANG II in renal microvessels by preventing activation of the large conductance Ca(2+-activated K(+ channel (KCa in vascular smooth muscle (VSM cells. ANG II increased the production of 20-HETE in rat renal microvessels. This response was attenuated by the 20-HETE synthesis inhibitors, 17-ODYA and HET0016, a phospholipase A2 inhibitor AACOF3, and the AT1 receptor blocker, Losartan, but not by the AT2 receptor blocker, PD123319. ANG II (10(-11 to 10(-6 M dose-dependently decreased the diameter of renal microvessels by 41 ± 5%. This effect was blocked by 17-ODYA. ANG II (10(-7 M did not alter KCa channel activity recorded from cell-attached patches on renal VSM cells under control conditions. However, it did reduce the NPo of the KCa channel by 93.4 ± 3.1% after the channels were activated by increasing intracellular calcium levels with ionomycin. The inhibitory effect of ANG II on KCa channel activity in the presence of ionomycin was attenuated by 17-ODYA, AACOF3, and the phospholipase C (PLC inhibitor U-73122. ANG II induced a peak followed by a steady-state increase in intracellular calcium concentration in renal VSM cells. 17-ODYA (10(-5 M had no effect on the peak response, but it blocked the steady-state increase. These results indicate that ANG II stimulates the formation of 20-HETE in rat renal microvessels via the AT1 receptor activation and that 20-HETE contributes to the vasoconstrictor response to ANG II by blocking activation of KCa channel and facilitating calcium entry.

  14. The concentration of plasma metabolites varies throughout reproduction and affects offspring number in wild brown trout (Salmo trutta).

    Gauthey, Zoé; Freychet, Marine; Manicki, Aurélie; Herman, Alexandre; Lepais, Olivier; Panserat, Stéphane; Elosegi, Arturo; Tentelier, Cédric; Labonne, Jacques

    2015-06-01

    In wild populations, measuring energy invested in the reproduction and disentangling investment in gametes versus investment in reproductive behavior (such as intrasexual competition or intersexual preference) remain challenging. In this study, we investigated the energy expenditure in brown trout reproductive behavior by using two proxies: variation in weight and variation of plasma metabolites involved in energy production, over the course of reproductive season in a semi natural experimental river. We estimated overall reproductive success using genetic assignment at the end of the reproductive season. Results show that triglycerides and free fatty acid concentrations vary negatively during reproduction, while amino-acids and glucose concentrations remain stable. Decrease in triglyceride and free fatty acid concentrations during reproduction is not related to initial concentration levels or to weight variation. Both metabolite concentration variations and weight variations are correlated to the number of offspring produced, which could indicate that gametic and behavioral reproductive investments substantially contribute to reproductive success in wild brown trout. This study opens a path to further investigate variations in reproductive investment in wild populations. PMID:25666363

  15. Analysis of naltrexone urinary metabolites.

    Ventura, R; de la Torre, R; Segura, J

    1988-01-01

    A reversed-phase HPLC method using ion-pair formation has been developed for the simultaneous determination of naltrexone and three urinary metabolites. The extraction of the free and conjugated metabolites was studied by liquid-solid procedures using styrene-divinylbenzene copolymers (Amberlite XAD-2) and bonded octadecyl silica supports (ODS-silica). Optimum recovery was obtained with ODS-silica extraction using 25% acetonitrile in a 5 mM diammonium phosphate buffer pH 2.1 as elution solvent. The chromatographic behaviour of naltrexone metabolites and naloxone (internal standard) was examined by varying the mobile phase composition. Increments of both the diammonium phosphate buffer concentration and the percentage of organic solvent in the eluent decreases the retention of compounds in a non-linear manner. Increments of the dodecyl sulphate (counter-ion) concentration, increases the retention time. The method was applied to determine the urinary levels of major naltrexone metabolites in a volunteer receiving a 50 mg oral dose. This is the first method reported which permits the simultaneous quantitative determination of naltrexone and its metabolites, 6beta-naltrexol, naltrexone glucuronide and 6beta-naltrexol glucuronide, in urine. PMID:16867358

  16. Contribution of post-harvest agricultural paddy residue fires in the N.W. Indo-Gangetic Plain to ambient carcinogenic benzenoids, toxic isocyanic acid and carbon monoxide

    Praphulla Chandra, Boggarapu; Sinha, Vinayak

    2016-04-01

    benzene and ensure compliance with the NAAQS. Calculations of excessive lifetime cancer risk due to benzene amount to 25 and 10 per million inhabitants for children and adults, respectively, exceeding the USEPA threshold of 1 per million inhabitants. Annual exposure to isocyanic acid was close to 1 ppb, the concentration considered to be sufficient to enhance risks for cardiovascular diseases and cataracts. This study makes a case for urgent mitigation of post-harvest paddy residue fires as the unknown synergistic effect of multi-pollutant exposure due to emissions from this anthropogenic source may be posing grave health risks to the population of the N.W. IGP. This work has been published very recently and the citation to the complete work is: B.P. Chandra, Vinayak Sinha, Contribution of post-harvest agricultural paddy residue fires in the N.W. Indo-Gangetic Plain to ambient carcinogenic benzenoids, toxic isocyanic acid and carbon monoxide, Environment International, Volume 88, March 2016, Pages 187-197, ISSN 0160-4120, http://dx.doi.org/10.1016/j.envint.2015.12.025.

  17. Contribution of Amino Acid Region 659−663 of Factor Va Heavy Chain to the Activity of Factor Xa within Prothrombinase†,‡

    Hirbawi, Jamila; John L. Vaughn; Bukys, Michael A.; Vos, Hans L.; Kalafatis, Michael

    2010-01-01

    Factor Va, the cofactor of prothrombinase, is composed of heavy and light chains associated noncovalently in the presence of divalent metal ions. The COOH-terminal region of the heavy chain contains acidic amino acid clusters that are important for cofactor activity. In this work, we have investigated the role of amino acid region 659−663, which contains five consecutive acidic amino acid residues, by site-directed mutagenesis. We have generated factor V molecules in which all residues were m...

  18. Determination of rifampicin and its main metabolite in plasma and urine in presence of pyrazinamide and isoniazid by HPLC method.

    Panchagnula, R; Sood, A; Sharda, N; Kaur, K; Kaul, C L

    1999-01-01

    A reversed phase HPLC method is described for the simultaneous estimation of rifampicin and its major metabolite desacetyl rifampicin, in the presence of isoniazid and pyrazinamide, in human plasma and urine. The assay involves simple liquid extraction of drug, metabolite and internal standard (rifapentine) from biological specimens and their subsequent separation on a C18 reversed phase column and single wavelength UV detection. In plasma as well as in urine samples, all the three compounds of interest eluted within 17 min. Using methanol-sodium phosphate buffer (pH 5.2; 0.01 M) (65:35, v/v) as mobile phase under isocratic conditions, it was established that isoniazid, pyrazinamide and ascorbic acid (added to prevent oxidative degradation of analytes) did not interfere with the analyte peaks. Recoveries (extraction efficiency) for drug were greater than 90% in both plasma and urine, whereas for metabolite the values were found to be 79 and 86% in plasma and urine, respectively. The plasma and urine methods were precise (total coefficient of variation ranged from 5 to 23%) and accurate (-7 to 5% of the nominal values) for both the analytes. Individual variance components, their estimates and their contribution to the total variance were also determined. Using the same method, unknown samples supplied by WHO were assayed and good correlations were obtained between the found and intended values. The method developed proved to be suitable for simultaneous estimation of rifampicin and desacetyl rifampicin in plasma and urine samples. PMID:9925337

  19. Glutamine and glutamate as vital metabolites

    Newsholme P.

    2003-01-01

    Full Text Available Glucose is widely accepted as the primary nutrient for the maintenance and promotion of cell function. This metabolite leads to production of ATP, NADPH and precursors for the synthesis of macromolecules such as nucleic acids and phospholipids. We propose that, in addition to glucose, the 5-carbon amino acids glutamine and glutamate should be considered to be equally important for maintenance and promotion of cell function. The functions of glutamine/glutamate are many, i.e., they are substrates for protein synthesis, anabolic precursors for muscle growth, they regulate acid-base balance in the kidney, they are substrates for ureagenesis in the liver and for hepatic and renal gluconeogenesis, they act as an oxidative fuel for the intestine and cells of the immune system, provide inter-organ nitrogen transport, and act as precursors of neurotransmitter synthesis, of nucleotide and nucleic acid synthesis and of glutathione production. Many of these functions are interrelated with glucose metabolism. The specialized aspects of glutamine/glutamate metabolism of different glutamine-utilizing cells are discussed in the context of glucose requirements and cell function.

  20. Antagonism of presynaptic dopamine receptors by phenothiazine drug metabolites

    Electrically evoked release of dopamine from the caudate nucleus is reduced by the dopamine receptor agonists, apomorphine and bromocriptine, and facilitated by neuroleptic drugs, which act as dopamine autoreceptor antagonists. The potencies of chlorpromazine, fluphenazine, levomepromazine and their hydroxy-metabolites in modulating electrically evoked release of dopamine were examined by superfusion of rabbit caudate nucleus slices pre-incubated with 3H-dopamine. O-Desmethyl levomepromazine, 3-hydroxy- and 7-hydroxy metabolites of chlorpromazine and levomepromazine facilitated electrically evoked release of 3H-dopamine, having potencies similar to that of the parent compounds. 7-Hydroxy fluphenazine was less active than fluphenazine in this system. These results indicate that phenolic metabolites of chlorpromazine and levomepromazine, but not of fluphenazine, may contribute to effects of the drugs mediated by presynaptic dopamine receptors

  1. In vivo MRS metabolite quantification using genetic optimization

    The in vivo quantification of metabolites' concentrations, revealed in magnetic resonance spectroscopy (MRS) spectra, constitutes the main subject under investigation in this work. Significant contributions based on artificial intelligence tools, such as neural networks (NNs), with good results have been presented lately but have shown several drawbacks, regarding their quantification accuracy under difficult conditions. A general framework that encounters the quantification procedure as an optimization problem, which is solved using a genetic algorithm (GA), is proposed in this paper. Two different lineshape models are examined, while two GA configurations are applied on artificial data. Moreover, the introduced quantification technique deals with metabolite peaks' overlapping, a considerably difficult situation occurring under real conditions. Appropriate experiments have proved the efficiency of the introduced methodology, in artificial MRS data, by establishing it as a generic metabolite quantification procedure

  2. A 20-amino-acid deletion in the neuraminidase stalk and a five-amino-acid deletion in the NS1 protein both contribute to the pathogenicity of H5N1 avian influenza viruses in mallard ducks.

    Yanfang Li

    Full Text Available Since 2003, H5N1-subtype avian influenza viruses (AIVs with both a deletion of 20 amino acids in the stalk of the neuraminidase (NA glycoprotein (A- and a deletion of five amino acids at positions 80 to 84 in the non-structural protein NS1 (S- have become predominant. To understand the influence of these double deletions in the NA and NS1 proteins on the pathogenicity of H5N1-subtype AIVs, we selected A/mallard/Huadong/S/2005 as a parental strain to generate rescued wild-type A-S- and three variants (A-S+ with a five-amino-acid insertion in the NS1 protein, A+S- with a 20-amino-acid insertion in the NA stalk, and A+S+ with insertions in both NA and NS1 proteins and evaluated their biological characteristics and virulence. The titers of the AIVs with A- and/or S- replicated in DEF cells were higher than that of A+S+, and the A-S- virus exhibited a replication predominance when co-infected with the other variants in DEF cells. In addition, A-S- induced a more significant increase in the expression of immune-related genes in peripheral blood mononuclear cells of mallard ducks in vitro compared with the other variants. Furthermore, an insertion in the NA and/or NS1 proteins of AIVs resulted in a notable decrease in virulence in ducks, as determined by intravenous pathogenicity index, and the two insertions exerted a synergistic effect on the attenuation of pathogenicity in ducks. In addition, compared with A+S+ and A+S-, the A-S+ and A-S- viruses that were introduced via the intranasal inoculation route exhibited a faster replication ability in the lungs of ducks. These data indicate that both the deletions in the NA stalk and the NS1 protein contribute to the high pathogenicity of H5N1 AIVs in ducks.

  3. Dynamics of intestinal metabolites and morphology in response to necrotic enteritis challenge in broiler chickens.

    Wu, Shu-Biao; Rodgers, Nicholas J; Cui, Guimei; Sun, Yi; Choct, Mingan

    2016-06-01

    Despite the relatively small contribution to metabolizable energy that volatile fatty acids (VFAs) provide in chickens, these organic acids have been reported to play beneficial roles in the gastrointestinal tract (GIT) of birds, for example, inhibition of the growth of some pathogenic bacteria. However, information regarding the dynamics of these metabolites in the GIT of chickens is still scarce, especially under disease conditions such as necrotic enteritis (NE). Here, we investigated the dynamics of VFAs and lactic acid, and intestinal morphology in response to NE predisposing factors, that is, excessive dietary fishmeal and Eimeria inoculation, and causative agent Clostridium perfringens producing NetB toxin. The experiment was designed in a 2 × 2 × 2 factorial arrangement of treatments with or without: fishmeal feeding, Eimeria inoculation and C. perfringens challenge. The results showed that these factors significantly influenced composition and concentration of VFAs and lactic acids, pH and histomorphometry in one way or another. These changes may be important for the onset of NE or only the synergetic responses to micro environmental stress. Eimeria appeared to be more important than fishmeal in predisposing birds to NE, thus the application of Eimeria in NE challenge provides more consistent success in inducing the disease. The metabolic responses to various adverse factors such as excessive dietary fishmeal and Eimeria infection are complex. Thus, intensive efforts are required to better understand NE so as to achieve the control of the disease in the absence of antibiotics. PMID:27245303

  4. Investigating associations between milk metabolite profiles and milk traits of Holstein cows.

    Melzer, N; Wittenburg, D; Hartwig, S; Jakubowski, S; Kesting, U; Willmitzer, L; Lisec, J; Reinsch, N; Repsilber, D

    2013-03-01

    In the field of dairy cattle research, it is of great interest to improve the detection and prevention of diseases (e.g., mastitis and ketosis) and monitor specific traits related to the state of health and management. During the standard milk performance test, traditional milk traits are monitored, and quality and quantity are screened. In addition to the standard test, it is also now possible to analyze milk metabolites in a high-throughput manner and to consider them in connection with milk traits to identify functionally important metabolites that can also serve as biomarker candidates. We present a study in which 190 milk metabolites and 14 milk traits of 1,305 Holstein cows on 18 commercial farms were investigated to characterize interrelations of milk metabolites between each other, to milk traits from the milk standard performance test, and to influencing factors such as farm and sire effect (half-sib structure). The effect of influencing factors (e.g., farm) varied among metabolites and traditional milk traits. The investigations of associations between metabolites and milk traits revealed groups of metabolites that show, for example, positive correlations to protein and casein, and negative correlations to lactose and pH. On the other hand, groups of metabolites jointly associated with the investigated milk traits can be identified and functionally discussed. To enable a multivariate investigation, 2 machine learning methods were applied to detect important metabolites that are highly correlated with the investigated traditional milk traits. For somatic cell score, uracil, lactic acid, and 9 other important metabolites were detected. Lactic acid has already been proposed as a biomarker candidate for mastitis in the recent literature. In conclusion, we found sets of metabolites eligible to predict milk traits, enabling the analysis of milk traits from a metabolic perspective and discussion of the possible functional background for some of the detected

  5. Tissue metabolite profiling identifies differentiating and prognostic biomarkers for prostate carcinoma.

    Jung, Klaus; Reszka, Regina; Kamlage, Beate; Bethan, Bianca; Stephan, Carsten; Lein, Michael; Kristiansen, Glen

    2013-12-15

    Metabolomic research offers a deeper insight into biochemical changes in cancer metabolism and is a promising tool for identifying novel biomarkers. We aimed to evaluate the diagnostic and prognostic potential of metabolites in prostate cancer (PCa) tissue after radical prostatectomy. In matched malignant and nonmalignant prostatectomy samples from 95 PCa patients, aminoadipic acid, cerebronic acid, gluconic acid, glycerophosphoethanolamine, 2-hydroxybehenic acid, isopentenyl pyrophosphate, maltotriose, 7-methylguanine and tricosanoic acid were determined within a global metabolite profiling study using gas chromatography/liquid chromatography-mass spectrometry. The data were related to clinicopathological variables like prostate volume, tumor stage, Gleason score, preoperative prostate-specific antigen and disease recurrence in the follow-up. All nine metabolites showed higher concentrations in malignant than in nonmalignant samples except for gluconic acid and maltotriose, which had lower levels in tumors. Receiver -operating characteristics analysis demonstrated a significant discrimination for all metabolites between malignant and nonmalignant tissue with a maximal area under the curve of 0.86 for tricosanoic acid, whereas no correlation was observed between the metabolite levels and the Gleason score or tumor stage except for gluconic acid. Univariate Cox regression and Kaplan-Meier analyses showed that levels of aminoadipic acid, gluconic acid and maltotriose were associated with the biochemical tumor recurrence (prostate-specific antigen > 0.2 ng/mL). In multivariate Cox regression analyses, aminoadipic acid together with tumor stage and Gleason score remained in a model as independent marker for prediction of biochemical recurrence. This study proved that metabolites in PCa tissue can be used, in combination with traditional clinicopathological factors, as promising diagnostic and prognostic tools. PMID:23737455

  6. Reactive Arrays of Colorimetric Sensors for Metabolite and Steroid Identification.

    Batres, Gary; Jones, Talia; Johnke, Hannah; Wilson, Mark; Holmes, Andrea E; Sikich, Sharmin

    2014-12-31

    The work described herein examines a rapid mix-and-measure method called DETECHIP suitable for screening of steroids and metabolites. The addition of steroids and metabolites to reactive arrays of colorimetric sensors generated characteristic color "fingerprints" that were used to identify the analyte. A color analysis tool was used to identify the analyte pool that now includes biologically relevant analytes. The mix-and-measure arrays allowed the detection of disease metabolites, orotic acid and argininosuccinic acid; and the steroids androsterone, 1,4-androstadiene, testosterone, stanozolol, and estrone. The steroid 1,4-androstadiene was also detected by this method while dissolved in synthetic urine. Some of the steroids, such as androstadiene, stanozolol, and androsterone were co-dissolved with (2-hydroxypropyl)-β-cyclodextrin in order to increase solubility in aqueous buffered solutions. The colorimetric arrays do not intend to eliminate ELISA or mass spectroscopy based screening, but to possibly provide an alternative analytical detection method for steroids and metabolites. PMID:25019034

  7. A Novel Fungal Metabolite with Beneficial Properties for Agricultural Applications

    Francesco Vinale

    2014-07-01

    Full Text Available Trichoderma are ubiquitous soil fungi that include species widely used as biocontrol agents in agriculture. Many isolates are known to secrete several secondary metabolites with different biological activities towards plants and other microbes. Harzianic acid (HA is a T. harzianum metabolite able to promote plant growth and strongly bind iron. In this work, we isolated from the culture filtrate of a T. harzianum strain a new metabolite, named isoharzianic acid (iso-HA, a stereoisomer of HA. The structure and absolute configuration of this compound has been determined by spectroscopic methods, including UV-Vis, MS, 1D and 2D NMR analyses. In vitro applications of iso-HA inhibited the mycelium radial growth of Sclerotinia sclerotiorum and Rhizoctonia solani. Moreover, iso HA improved the germination of tomato seeds and induced disease resistance. HPLC-DAD experiments showed that the production of HA and iso HA was affected by the presence of plant tissue in the liquid medium. In particular, tomato tissue elicited the production of HA but negatively modulated the biosynthesis of its analogue iso-HA, suggesting that different forms of the same Trichoderma secondary metabolite have specific roles in the molecular mechanism regulating the Trichoderma plant interaction.

  8. Energy Metabolism Disorder as a Contributing Factor of Rheumatoid Arthritis: A Comparative Proteomic and Metabolomic Study.

    Xin Yu Yang

    Full Text Available To explore the pathogenesis of rheumatoid arthritis (RA, the different metabolites were screened in synovial fluid by metabolomics.Synovial fluid from 25 RA patients and 10 normal subjects were analyzed by GC/TOF MS analysis so as to give a broad overview of synovial fluid metabolites. The metabolic profiles of RA patients and normal subjects were compared using multivariate statistical analysis. Different proteins were verified by qPCR and western blot. Different metabolites were verified by colorimetric assay kit in 25 inactive RA patients, 25 active RA patients and 20 normal subjects. The influence of hypoxia-inducible factor (HIF-1α pathway on catabolism was detected by HIF-1α knockdown.A subset of 58 metabolites was identified, in which the concentrations of 7 metabolites related to energy metabolism were significantly different as shown by importance in the projection (VIP (VIP ≥ 1 and Student's t-test (p<0.05. In the 7 metabolites, the concentration of glucose was decreased, and the concentration of lactic acid was increased in the synovial fluid of RA patients than normal subjects verified by colorimetric assay Kit. Receiver operator characteristic (ROC analysis shows that the concentration of glucose and lactic acid in synovial fluid could be used as dependable biomarkers for the diagnosis of active RA, provided an AUC of 0.906 and 0.922. Sensitivity and specificity, which were determined by cut-off points, reached 84% and 96% in sensitivity and 95% and 85% in specificity, respectively. The verification of different proteins identified in our previous proteomic study shows that the enzymes of anaerobic catabolism were up-regulated (PFKP and LDHA, and the enzymes of aerobic oxidation and fatty acid oxidation were down-regulated (CS, DLST, PGD, ACSL4, ACADVL and HADHA in RA patients. The expression of HIF-1α and the enzymes of aerobic oxidation and fatty acid oxidation were decreased and the enzymes of anaerobic catabolism were

  9. Natural products: Hunting microbial metabolites

    Schmidt, Eric W.

    2015-05-01

    Symbiotic bacteria synthesize many specialized small molecules; however, establishing the role these chemicals play in human health and disease has been difficult. Now, the chemical structure and mechanism of the Escherichia coli product colibactin provides insight into the link between this secondary metabolite and colorectal cancer.

  10. Primary expectations of secondary metabolites

    Plant secondary metabolites (e.g., phenolics) are important for human health, in addition to the organoleptic properties they impart to fresh and processed foods. Consumer expectations such as appearance, taste, or texture influence their purchasing decisions. Thorough identification of phenolic com...

  11. The secondary metabolite bioinformatics portal

    Weber, Tilmann; Kim, Hyun Uk

    2016-01-01

    . In this context, this review gives a summary of tools and databases that currently are available to mine, identify and characterize natural product biosynthesis pathways and their producers based on ‘omics data. A web portal called Secondary Metabolite Bioinformatics Portal (SMBP at http...

  12. Zinc-chelation contributes to the anti-angiogenic effect of ellagic acid on inhibiting MMP-2 activity, cell migration and tube formation.

    Sheng-Teng Huang

    Full Text Available BACKGROUND: Ellagic acid (EA, a dietary polyphenolic compound, has been demonstrated to exert anti-angiogenic effect but the detailed mechanism is not yet fully understood. The aim of this study was to investigate whether the zinc chelating activity of EA contributed to its anti-angiogenic effect. METHODS AND PRINCIPAL FINDINGS: The matrix metalloproteinases-2 (MMP-2 activity, a zinc-required reaction, was directly inhibited by EA as examined by gelatin zymography, which was reversed dose-dependently by adding zinc chloride. In addition, EA was demonstrated to inhibit the secretion of MMP-2 from human umbilical vein endothelial cells (HUVECs as analyzed by Western blot method, which was also reversed by the addition of zinc chloride. Reversion-inducing cysteine-rich protein with Kazal motifs (RECK, known to down-regulate the MMP-2 activity, was induced by EA at both the mRNA and protein levels which was correlated well with the inhibition of MMP-2 activity. Interestingly, zinc chloride could also abolish the increase of EA-induced RECK expression. The anti-angiogenic effect of EA was further confirmed to inhibit matrix-induced tube formation of endothelial cells. The migration of endothelial cells as analyzed by transwell filter assay was suppressed markedly by EA dose-dependently as well. Zinc chloride could reverse these two effects of EA also in a dose-dependent manner. Since magnesium chloride or calcium chloride could not reverse the inhibitory effect of EA, zinc was found to be involved in tube formation and migration of vascular endothelial cells. CONCLUSIONS/SIGNIFICANCE: Together these results demonstrated that the zinc chelation of EA is involved in its anti-angiogenic effects by inhibiting MMP-2 activity, tube formation and cell migration of vascular endothelial cells. The role of zinc was confirmed to be important in the process of angiogenesis.

  13. Understanding Boswellia papyrifera tree secondary metabolites through bark spectral analysis

    Girma, Atkilt; Skidmore, Andrew K.; de Bie, C. A. J. M.; Bongers, Frans

    2015-07-01

    Decision makers are concerned whether to tap or rest Boswellia Papyrifera trees. Tapping for the production of frankincense is known to deplete carbon reserves from the tree leading to production of less viable seeds, tree carbon starvation and ultimately tree mortality. Decision makers use traditional experience without considering the amount of metabolites stored or depleted from the stem-bark of the tree. This research was designed to come up with a non-destructive B. papyrifera tree metabolite estimation technique relevant for management using spectroscopy. The concentration of biochemicals (metabolites) found in the tree bark was estimated through spectral analysis. Initially, a random sample of 33 trees was selected, the spectra of bark measured with an Analytical Spectral Device (ASD) spectrometer. Bark samples were air dried and ground. Then, 10 g of sample was soaked in Petroleum ether to extract crude metabolites. Further chemical analysis was conducted to quantify and isolate pure metabolite compounds such as incensole acetate and boswellic acid. The crude metabolites, which relate to frankincense produce, were compared to plant properties (such as diameter and crown area) and reflectance spectra of the bark. Moreover, the extract was compared to the ASD spectra using partial least square regression technique (PLSR) and continuum removed spectral analysis. The continuum removed spectral analysis were performed, on two wavelength regions (1275-1663 and 1836-2217) identified through PLSR, using absorption features such as band depth, area, position, asymmetry and the width to characterize and find relationship with the bark extracts. The results show that tree properties such as diameter at breast height (DBH) and the crown area of untapped and healthy trees were strongly correlated to the amount of stored crude metabolites. In addition, the PLSR technique applied to the first derivative transformation of the reflectance spectrum was found to estimate the

  14. Towards a new method for the quantification of metabolites in the biological sample

    The quantification of metabolites is a key step in drug development. The aim of this Ph.D. work was to study the feasibility of a new method for this quantification, in the biological sample, without the drawbacks (cost, time, ethics) of the classical quantification methods based on metabolites synthesis or administration to man of the radiolabelled drug. Our strategy consists in determining the response factor, in mass spectrometry, of the metabolites. This approach is based on tritium labelling of the metabolites, ex vivo, by isotopic exchange. The labelling step was studied with deuterium. Metabolites of a model drug, recovered from in vitro or urinary samples, were labelled by three ways (Crab tree's catalyst ID2, deuterated trifluoroacetic acid or rhodium chloride ID20). Then, the transposition to tritium labelling was studied and the first results are very promising for the ultimate validation of the method. (author)

  15. The use of secondary metabolite profiling in chemotaxonomy of filamentous fungi

    Frisvad, Jens Christian; Andersen, Birgitte; Thrane, Ulf

    2008-01-01

    A secondary metabolite is a chemical compound produced by a limited number of fungal species in a genus, an order, or even phylum. A profile of secondary metabolites consists of all the different compounds a fungus can produce on a given substratum and includes toxins, antibiotics and other outward......-directed compounds. Chemotaxonomy is traditionally restricted to comprise fatty acids, proteins, carbohydrates, or secondary metabolites, but has sometimes been defined so broadly that it also includes DNA sequences. It is not yet possible to use secondary metabolites in phylogeny, because of the inconsistent...... have been analysed for a wide array of terpenes, polyketides, non-ribosomal peptides, and combinations of these. Fungal chemotaxonomy based on secondary metabolites has been used successfully in large ascomycete genera such as Alternaria, Aspergillus, Fusarium, Hypoxylon, Penicillium, Stachybotrys...

  16. Identification of Epoxide-Derived Metabolite(s) of Benzbromarone.

    Wang, Kai; Wang, Hui; Peng, Ying; Zheng, Jiang

    2016-04-01

    Benzbromarone (BBR) is a benzofuran derivative that has been quite useful for the treatment of gout; however, it was withdrawn from European markets in 2003 because of reported serious incidents of drug-induced liver injury. BBR-induced hepatotoxicity has been suggested to be associated with the formation of a quinone intermediate. The present study reported epoxide-derived intermediate(s) of BBR. An N-acetylcysteine (NAC) conjugate derived from epoxide metabolite(s) was detected in both microsomal incubations of BBR and urine samples of mice treated with BBR. The NAC conjugate was identified as 6-NAC BBR. Ketoconazole suppressed the bioactivation of BBR to the epoxide intermediate(s), and the CYP3A subfamily was the primary enzyme responsible for the formation of the epoxide(s). The present study provided new information on metabolic activation of BBR. PMID:26792818

  17. Targeted metabolomics identifies reliable and stable metabolites in human serum and plasma samples.

    Michaela Breier

    Full Text Available BACKGROUND: Information regarding the variability of metabolite levels over time in an individual is required to estimate the reproducibility of metabolite measurements. In intervention studies, it is critical to appropriately judge changes that are elicited by any kind of intervention. The pre-analytic phase (collection, transport and sample processing is a particularly important component of data quality in multi-center studies. METHODS: Reliability of metabolites (within-and between-person variance, intraclass correlation coefficient and stability (shipment simulation at different temperatures, use of gel-barrier collection tubes, freeze-thaw cycles were analyzed in fasting serum and plasma samples of 22 healthy human subjects using a targeted LC-MS approach. RESULTS: Reliability of metabolite measurements was higher in serum compared to plasma samples and was good in most saturated short-and medium-chain acylcarnitines, amino acids, biogenic amines, glycerophospholipids, sphingolipids and hexose. The majority of metabolites were stable for 24 h on cool packs and at room temperature in non-centrifuged tubes. Plasma and serum metabolite stability showed good coherence. Serum metabolite concentrations were mostly unaffected by tube type and one or two freeze-thaw cycles. CONCLUSION: A single time point measurement is assumed to be sufficient for a targeted metabolomics analysis of most metabolites. For shipment, samples should ideally be separated and frozen immediately after collection, as some amino acids and biogenic amines become unstable within 3 h on cool packs. Serum gel-barrier tubes can be used safely for this process as they have no effect on concentration in most metabolites. Shipment of non-centrifuged samples on cool packs is a cost-efficient alternative for most metabolites.

  18. Functional antidopaminergic and anticholinergic effects of thioridazine and its metabolites

    Niedzwiecki, D.

    1986-01-01

    The relative potency of thioridazine and two of its clinically active metabolites, mesoridazine and sulforidazine was studied. In each of three separate methods, mesoridazine and sulforidazine exhibited greater potency than did thioridazine. Both metabolites showed greater affinities for striatal DA receptors as estimated by their competition for (/sup 3/H)spiperone binding sites in crude striatal membrane preparations. On a more functional level, both metabolites more potently antagonized the inhibitory effects of either the direct acting DA agonist apomorphine, or of endogenous DA, on the electrically evoked release of radiolabeled DA and ACh from perfused striatal slices. While thioridazine effectively blocked the agonist-induced inhibition at those striatal DA receptors which control DA release, it showed significantly lower potency at the striatal DA receptors which modulate ACh release. Thioridazine exhibited moderate affinity for striatal muscarinic cholinergic receptors. It was only five times less potent than atropine in competing for (/sup 3/H) quinuclidinylbenzilate binding sites in striatal membrane preparations. Unlike dopamine receptors, thioridazine showed greater antimuscarinic potency than did its metabolites. Despite this significant affinity for muscarinic binding sites in striatal homogenates, neither thioridezine nor its metabolites could block the inhibitory effects of the full muscarinic agonist carbachol, on the evoked release of ACh from striatal slices. This lack of effect contrasted with the antagonism of the carbachol-induced inhibition by such classical muscarinic blockers as quinuclindinylbenzilate or atropine. In combination with available pharmacokinetic data, these studies have demonstrated that the metabolites of thioridazine probably contribute to the antidopaminergic effects of this drug within the CNS.

  19. Profiling of metabolites in oil palm mesocarp at different stages of oil biosynthesis.

    Neoh, Bee Keat; Teh, Huey Fang; Ng, Theresa Lee Mei; Tiong, Soon Huat; Thang, Yin Mee; Ersad, Mohd Amiron; Mohamed, Mohaimi; Chew, Fook Tim; Kulaveerasingam, Harikrishna; Appleton, David R

    2013-02-27

    Oil palm is one of the most productive oil producing crops and can store up to 90% oil in its fruit mesocarp. However, the biosynthetic regulation and drivers of palm mesocarp development are still not well understood. Multiplatform metabolomics technology was used to profile palm metabolites during six critical stages of fruit development in order to better understand lipid biosynthesis. Significantly higher amino acid levels were observed in palm mesocarp preceding lipid biosynthesis. Nucleosides were found to be in high concentration during lipid biosynthesis, whereas levels of metabolites involved in the tricarboxylic acid cycle were more concentrated during early fruit development. Apart from insights into the regulation of metabolites during fruit development in oil palm, these results provide potentially useful metabolite yield markers and genes of interest for use in breeding programs. PMID:23384169

  20. Contribution of carbohydrate and amino acids to the formation of aromatic structure of synthetic melanoidin (model sedimentary humic substance). A study using 13C-labeled glucose

    As a part of study to elucidate the source of aromatic structure in sedimentary humic substances (SHS), we synthesized melanoidins (a model SHS) from 13C-labeled glucose and non-labeled amino acids and examined how glucose and amino acids are involved in the formation of their aromatic structures. The synthesized melanoidin was oxidized by alkaline perman ganate and benzenecarboxylic acids in their degradation products were analyzed by Chemical Ionization-GC/MS. The results indicate that aromatic structures are formed in melanoidin as a result of both selfcondensation of glucose and a reaction between glucose and amino acids. (author)

  1. A contribution to the distinction of biogenic vinegar and vinegar made from synthetic acetic acid by determining the specific 14C-radioactivity

    The method of Simon et al. for the separation of the acetic acid from vinegar prior to the determination of the specific 14C-radioactivity has been modified. The precipitation as calcium acetate and the preparation of free acetic acid by addition of diphosphoric acid has been replaced by an extraction procedure with diisopropylether which is faster and cheaper. On the Austrian market glacial acetic acid (Merck, p.A.) having the natural specific 14C-radioactivity was found. The natural specific 14C-radioactivity is therefore necessary but not sufficient to prove the biogenic origin of vinegar. (orig.)

  2. Differential metabolite profiles during fruit development in high-yielding oil palm mesocarp.

    Huey Fang Teh

    Full Text Available To better understand lipid biosynthesis in oil palm mesocarp, in particular the differences in gene regulation leading to and including de novo fatty acid biosynthesis, a multi-platform metabolomics technology was used to profile mesocarp metabolites during six critical stages of fruit development in comparatively high- and low-yielding oil palm populations. Significantly higher amino acid levels preceding lipid biosynthesis and nucleosides during lipid biosynthesis were observed in a higher yielding commercial palm population. Levels of metabolites involved in glycolysis revealed interesting divergence of flux towards glycerol-3-phosphate, while carbon utilization differences in the TCA cycle were proven by an increase in malic acid/citric acid ratio. Apart from insights into the regulation of enhanced lipid production in oil palm, these results provide potentially useful metabolite yield markers and genes of interest for use in breeding programmes.

  3. Differential metabolite profiles during fruit development in high-yielding oil palm mesocarp.

    Teh, Huey Fang; Neoh, Bee Keat; Hong, May Ping Li; Low, Jaime Yoke Sum; Ng, Theresa Lee Mei; Ithnin, Nalisha; Thang, Yin Mee; Mohamed, Mohaimi; Chew, Fook Tim; Yusof, Hirzun Mohd; Kulaveerasingam, Harikrishna; Appleton, David R

    2013-01-01

    To better understand lipid biosynthesis in oil palm mesocarp, in particular the differences in gene regulation leading to and including de novo fatty acid biosynthesis, a multi-platform metabolomics technology was used to profile mesocarp metabolites during six critical stages of fruit development in comparatively high- and low-yielding oil palm populations. Significantly higher amino acid levels preceding lipid biosynthesis and nucleosides during lipid biosynthesis were observed in a higher yielding commercial palm population. Levels of metabolites involved in glycolysis revealed interesting divergence of flux towards glycerol-3-phosphate, while carbon utilization differences in the TCA cycle were proven by an increase in malic acid/citric acid ratio. Apart from insights into the regulation of enhanced lipid production in oil palm, these results provide potentially useful metabolite yield markers and genes of interest for use in breeding programmes. PMID:23593468

  4. Identification of some benproperine metabolites in humans and investigation of their antitussive effect

    Yan LI; Da-fang ZHONG; Si-wei CHEN; Isamu MAEBA

    2005-01-01

    Aim: To identify 4 unknown metabolites of benproperine (BPP, 1) in human urine after a po dose, and to investigate the antitussive effect of monohydroxylate metabolites. Methods: The putative metabolite references were prepared using chemical synthesis. Their structures were identified using 1H and 13C nuclear magnetic resonance, and mass spectrometry. The metabolites in human urine were separated and assayed using liquid chromatography-ion trap mass spectrometry (LC/MS/MS), and further confirmed by comparison of their mass spectra and chromatographic retention times with those of synthesized reference substances. The antitussive effects of metabolites were evaluated on coughs induced by 7.5% citric acid in conscious guinea pigs. Results: 1-[1-Methyl-2-[2-(phenylmethyl)phenoxy] -ethyl] -4-piperidinol (2), 1- [ 1-methyl-2- [2-(phenylmethyl)phenoxy] ethyl]-3-piperidinol (3) and their glucuronides 4 and 5 were obtained from chemical synthesis. Four urinary metabolites in human urine showed peaks with the same chromatographic retention times and mass spectra in LC/MS/MS as synthetic substances 2, 3, 4 and 5. Phosphates of compounds 2 and 3 prolonged the latency of cough and reduced the number of coughs during the 3 min test using citric acid, but did not reduce the number of coughs during the 5 min immediately after the test in conscious guinea pigs. Conclusion: Compounds 2, 3, 4,and 5 were identified as the metabolites of BPP in human urine. Among them,compounds 2 and 3 are inactive in the antitussive effect.

  5. Two Polycyclic Geranylhydroquinone-Derived Metabolites from Roots of Arnebia hispidissima (Lehm. DC.

    Atallah F. Ahmed

    2014-05-01

    Full Text Available A phytochemical investigation of the least polar organic extracts of Arnebia hispidissima (Lehm. DC. roots has led to the isolation of two unique polycyclic geranylhydroquinone-derived metabolites, arnebacene (1 and arnebidin (2, along with some known phenolic metabolites viz., arnebin-7 (3 and vanillic acid (4. The chemical identification of the new isolated compounds, including their relative stereochemistry, was achieved via spectroscopic analyses, including 2D NMR, and spectral comparison with related compounds. A biosynthetic pathway is proposed for the new compounds on the basis of their structure-relationship with previously isolated metabolites.

  6. Fusarium verticillioides SGE1 is required for full virulence and regulates expression of protein effector and secondary metabolite biosynthetic genes.

    Brown, Daren W; Busman, Mark; Proctor, Robert H

    2014-08-01

    The transition from one lifestyle to another in some fungi is initiated by a single orthologous gene, SGE1, that regulates markedly different genes in different fungi. Despite these differences, many of the regulated genes encode effector proteins or proteins involved in the synthesis of secondary metabolites (SM), both of which can contribute to pathogenicity. Fusarium verticillioides is both an endophyte and a pathogen of maize and can grow as a saprophyte on dead plant material. During growth on live maize plants, the fungus can synthesize a number of toxic SM, including fumonisins, fusarins, and fusaric acid, that can contaminate kernels and kernel-based food and feed. In this study, the role of F. verticillioides SGE1 in pathogenicity and secondary metabolism was examined by gene deletion analysis and transcriptomics. SGE1 is not required for vegetative growth or conidiation but is required for wild-type pathogenicity and affects synthesis of multiple SM, including fumonisins and fusarins. Induced expression of SGE1 enhanced or reduced expression of hundreds of genes, including numerous putative effector genes that could contribute to growth in planta; genes encoding cell surface proteins; gene clusters required for synthesis of fusarins, bikaverin, and an unknown metabolite; as well as the gene encoding the fumonisin cluster transcriptional activator. Together, our results indicate that SGE1 has a role in global regulation of transcription in F. verticillioides that impacts but is not absolutely required for secondary metabolism and pathogenicity on maize. PMID:24742071

  7. Transport of the soy isoflavone daidzein and its conjugative metabolites by the carriers SOAT, NTCP, OAT4, and OATP2B1.

    Grosser, Gary; Döring, Barbara; Ugele, Bernhard; Geyer, Joachim; Kulling, Sabine E; Soukup, Sebastian T

    2015-12-01

    Soy isoflavones (IF) are phytoestrogens, which interact with estrogen receptors. They are extensively metabolized by glucuronosyltransferases and sulfotransferases, leading to the modulation of their estrogenic activity. It can be assumed that this biotransformation also has a crucial impact on the uptake of IF by active or passive cellular transport mechanisms, but little is known about the transport of IF phase II metabolites into the cell. Therefore, transport assays for phase II metabolites of daidzein (DAI) were carried out using HEK293 cell lines transfected with five human candidate carriers, i.e., organic anion transporter OAT4, sodium-dependent organic anion transporter (SOAT), Na(+)-taurocholate cotransporting polypeptide (NTCP), apical sodium-dependent bile acid transporter ASBT, and organic anion transporting polypeptide OATP2B1. Cellular uptake was monitored by UHPLC-DAD. DAI monosulfates were transported by the carriers NTCP and SOAT in a sodium-dependent manner, while OAT4-HEK293 cells revealed a partly sodium-dependent transport for these compounds. In contrast, DAI-7,4'-disulfate was only taken up by NTCP-HEK293 cells. DAI-7-glucuronide, but not DAI-4'-glucuronide, was transported exclusively by OATP2B1 in a sodium-independent manner. DAI-7-glucuronide-4'-sulfate, DAI-7-glucoside, and DAI were no substrate of any of the tested carriers. In addition, the inhibitory potency of the DAI metabolites toward estrone-sulfate (E1S) uptake of the above-mentioned carriers was determined. In conclusion, human SOAT, NTCP, OATP2B1, and OAT4 were identified as carriers for the DAI metabolites. Several metabolites were able to inhibit carrier-dependent E1S uptake. These findings might contribute to a better understanding of the bioactivity of IF especially in case of hormone-related cancers. PMID:25319728

  8. Characterization of oxygenated metabolites of ginsenoside Rg1 in plasma and urine of rat.

    Wang, Jing-Rong; Tong, Tian-Tian; Yau, Lee-Fong; Chen, Cheng-Yu; Bai, Li-Ping; Ma, Jing; Hu, Ming; Liu, Liang; Jiang, Zhi-Hong

    2016-07-15

    This study describes the characterization of oxygenated metabolites of ginsenoside Rg1 in rat urine and plasma. These in vivo metabolites were profiled by using UHPLC-QTOF MS-based method. On the basis of high-resolution MS/MS data, and comparison with chemically synthesized authentic compounds, nine oxygenated metabolites of Rg1 were characterized as vinaginsenosides 21 and 22 (M1 and M2), vinaginsenoside R15 (M3), 6-O-(β-d-glucopyranosyl)-20-O-(β-d-glucopyranosyl) 3β, 6α, 12β, 20(S)-tetrahydroxy-24ξ-hydroxydammar-25-ene (M4 and M5), floralginsenoside A (M7 and M8), floralginsenoside B (M9) and epoxyginsenoside Rg1 (M13), respectively. Among these metabolites, M4, M5 and M13 are new ginsenosides and others were detected as in vivo metabolites of Rg1 for the first time. In addition, a series of oxygenated metabolites of Rh1 and deglycosylated metabolite of Rg1, were observed and characterized by comparing with compounds synthesized by us, which revealed an association between C-20 configuration and the extent of oxidation metabolism. Appearance of all these metabolites in blood stream and urine after i.v. dosing and oral administration of Rg1 was further examined, which clearly showed that mono-oxygenated metabolites of Rg1 were major circulating metabolites at the early stage after dosing. Characterization of exact chemical structures of these circulating metabolites contribute greatly to our understanding of chemical exposure after consumption of ginseng products, and provide valuable information for explaining multiple bioactivities of ginseng products. PMID:26809375

  9. Isolation of an Glycyrrhizic Acid-producing Andophytic Fungus from Licorice and Analysis of Metabolites%一株产甘草酸内生真菌的分离及代谢产物分析

    王红霞; 李雅丽

    2011-01-01

    采用组织块法从采自内蒙古鄂尔多斯市的甘草根部分离纯化内生真菌,经液体发酵培养,抽提发酵粗产物,以甘草酸为标准品采用LC-MS法和HPLC法对这些真菌的代谢产物进行筛选,获得1株产甘草酸的内生菌,通过形态学研究初步确定该菌株为镰孢霉属.%A strain of glycyrrhizic acid-producing endophytic fungi was obtained from the rats of licorice in Erdos city of the Inner Mongolia autonomous region by tissue culture method. Then the purified endophytic fungal strains were fermented. The glycyrrhizic acid in the fungal extract was extract and confirmed by HPLC and LC-MS by comparison with glycyrrhizic acid standard. The strain B12 that was found as the glycyrrhizic acid-producing andophytic fungus was grouped into Fusarium based on the morphological traits.

  10. Rapid analysis of fungal cultures and dried figs for secondary metabolites by LC/TOF-MS

    A liquid chromatography-time-of-flight mass spectrometry (LC/TOF-MS) method has been developed for profiling fungal metabolites. The performance of the procedure in terms of mass accuracy, selectivity (specificity) and repeatability was established by spiking aflatoxins, ochratoxins, trichothecenes and other metabolites into blank growth media. After extracting, and carrying out LC/TOF-MS analysis, the standards were correctly identified by searching a specially constructed database of 465 secondary metabolites. To demonstrate the viability of this approach 11 toxigenic and four non-toxigenic fungi from reference collections were grown on various media, for 7-14 days. The method was also applied to two toxigenic fungi, A. flavus (200-138) and A. parasiticus (2999-465) grown on gamma radiation sterilised dried figs, for 7-14 days. The fungal hyphae plus a portion of growth media or portions of dried figs were solvent extracted and analysed by LC/TOF-MS using a rapid resolution microbore LC column. Data processing based on cluster analysis, showed that electrospray ionization (ESI)-TOF-MS could be used to unequivocally identify metabolites in crude extracts. Using the elemental metabolite database, it was demonstrated that from culture collection isolates, anticipated metabolites. The speed and simplicity of the method has meant that levels of these metabolites could be monitored daily in sterilised figs. Over a 14-day period, levels of aflatoxins and kojic acid maximised at 5-6 days, whilst levels of 5-methoxysterigmatocystin remained relatively constant. In addition to the known metabolites expected to be produced by these fungi, roquefortine A, fumagillin, fumigaclavine B, malformins (peptides), aspergillic acid, nigragillin, terrein, terrestric acid and penicillic acid were also identified

  11. Marine-Derived Metabolites of S-Adenosylmethionine as Templates for New Anti-Infectives

    Janice R. Sufrin

    2009-08-01

    Full Text Available S-Adenosylmethionine (AdoMet is a key biochemical co-factor whose proximate metabolites include methylated macromolecules (e.g., nucleic acids, proteins, phospholipids, methylated small molecules (e.g., sterols, biogenic amines, polyamines (e.g., spermidine, spermine, ethylene, and N-acyl-homoserine lactones. Marine organisms produce numerous AdoMet metabolites whose novel structures can be regarded as lead compounds for anti-infective drug design.

  12. Aleuria aurantia - indole metabolites of fruit bodies, mycelial culture and culture medium

    Janina Węgiel

    2014-08-01

    Full Text Available The aim of present study was to investigate and compare indole metabolites of fruit bodies, mycelium cultivated in vitro and culture medium of the fungus Aleuria aurantia (Fr. Fuck. By use of a number of chromatographic and spectroscopic methods several indole metabolites have been detected and identified among other the 3-indolebutyric acid was produced and extracted to the culture medium. Furthermore 3-indoleatonitrile and tryptophane degradative products have been found both in fruit bodies and mycelium.

  13. Aleuria aurantia - indole metabolites of fruit bodies, mycelial culture and culture medium

    Janina Węgiel; Stanisław Kohlmünzer

    2014-01-01

    The aim of present study was to investigate and compare indole metabolites of fruit bodies, mycelium cultivated in vitro and culture medium of the fungus Aleuria aurantia (Fr.) Fuck. By use of a number of chromatographic and spectroscopic methods several indole metabolites have been detected and identified among other the 3-indolebutyric acid was produced and extracted to the culture medium. Furthermore 3-indoleatonitrile and tryptophane degradative products have been found both in fruit bodi...

  14. Secondary Metabolites from Rubiaceae Species

    Daiane Martins; Cecilia Veronica Nunez

    2015-01-01

    This study describes some characteristics of the Rubiaceae family pertaining to the occurrence and distribution of secondary metabolites in the main genera of this family. It reports the review of phytochemical studies addressing all species of Rubiaceae, published between 1990 and 2014. Iridoids, anthraquinones, triterpenes, indole alkaloids as well as other varying alkaloid subclasses, have shown to be the most common. These compounds have been mostly isolated from the genera Uncaria, Psych...

  15. Autism and Phthalate Metabolite Glucuronidation

    Stein, T. Peter; Schluter, Margaret D.; Steer, Robert A.; Ming, Xue

    2013-01-01

    Exposure to environmental chemicals may precipitate autism spectrum disorders (ASD) in genetically susceptible children. Differences in the efficiency of the glucuronidation process may substantially modulate substrate concentrations and effects. To determine whether the efficiency of this pathway is compromised in children with ASD, we measured the efficiency of glucuronidation for a series of metabolites derived from the commonly used plasticizer, diethylhexyl phthalate. Spot urines were co...

  16. Epigenome targeting by probiotic metabolites

    Licciardi Paul V; Wong Sook-San; Tang Mimi LK; Karagiannis Tom C

    2010-01-01

    Abstract Background The intestinal microbiota plays an important role in immune development and homeostasis. A disturbed microbiota during early infancy is associated with an increased risk of developing inflammatory and allergic diseases later in life. The mechanisms underlying these effects are poorly understood but are likely to involve alterations in microbial production of fermentation-derived metabolites, which have potent immune modulating properties and are required for maintenance of...

  17. Recent advances in the analysis of the site-specific isotopic fractionation of metabolites such as fatty acids using anisotropic natural-abundance 2H NMR spectroscopy: application to conjugated linolenic methyl esters.

    Lesot, Philippe; Serhan, Zeinab; Billault, Isabelle

    2011-01-01

    The full elucidation of the enzymatic mechanisms leading to polyunsaturated ω-3 to ω-5 fatty acids (PUFAs) occurring in plants or microorganisms by analyzing their site-specific isotopic fractionation profiles is a challenging task. Isotropic SNIF-NMR® method is an historical, powerful tool for the determination of ((2)H/(1)H) ratios. However, the absence of accessible isotopic data on the enantiotopic hydrogen sites (CH(2) groups) prevents the study of the enzymatic reaction stereoselectivity. Natural-abundance deuterium (NAD) 2D NMR experiment using chiral liquid crystals (CLC) as solvent is a new tool in this field, overcoming this limitation. In this work, we have explored various possibilities for optimizing the enantio-discrimination properties of CLC by changing the nature of the polypeptide and/or increasing the polarity of the organic co-solvents. We report also the first applications of TMU as co-solvent for preparing enantio-discriminating, homogenous polypeptide mesophases. The various experimental NAD NMR results recorded at an optimal sample temperature are discussed and compared in terms of number of discriminated (2)H sites and magnitude of spectral separation for different PUFAs such as the linoleic and linolenic acids. The comparison of all NMR results shows that optimal results are obtained when CLC mixtures made of poly-γ-benzyl-L-glutamate (PBLG) and high polarity co-solvents are used. As new challenging examples of applications, we report the preliminary analytical results obtained from two ω-5 conjugated linolenic acids: the α-eleostearic acid (9Z, 11E, 13E) and the punicic acid (9Z, 11E, 13Z). NMR data are discussed in terms of molecular orientational ordering parameters and isotopic distribution. PMID:21107978

  18. Bioactive Metabolites from Spilanthes acmella Murr.

    Virapong Prachayasittikul

    2009-02-01

    Full Text Available Spilanthes acmella Murr. (Compositae has been used as a traditional medicine for toothache, rheumatism and fever. Its extracts had been shown to exhibit vasorelaxant and antioxidant activities. Herein, its antimicrobial, antioxidant and cytotoxic activities were evaluated. Agar dilution method assays against 27 strains of microorganisms were performed. Results showed that fractions from the chloroform and methanol extracts inhibited the growth of many tested organisms, e.g. Corynebacterium diphtheriae NCTC 10356 with minimum inhibitory concentration (MIC of 64-256 mg/mL and Bacillus subtilis ATCC 6633 with MIC of 128-256 mg/mL. The tested fractions all exhibited antioxidant properties in both DPPH and SOD assays. Potent radical scavenging activity was observed in the DPPH assay. No cytotoxic effects of the extracts against KB and HuCCA-1 cell lines were evident. Bioassay-guided isolation resulted in a diverse group of bioactive compounds such as phenolics [vanillic acid (2, trans-ferulic acid (5 and trans-isoferulic acid (6], coumarin (scopoletin, 4 and triterpenoids like 3-acetylaleuritolic acid (1, b-sitostenone (3, stigmasterol and stigmasteryl-3-O-b-D-glucopyranosides, in addition to a mixture of stigmasteryl-and b-sitosteryl-3-O-b-D-glucopyranosides. The compounds 1–6 represent bioactive metabolites of S. acmella Murr. that were never previously reported. Our findings demonstrate for the first time the potential benefits of this medicinal plant as a rich source of high therapeutic value compounds for medicines, cosmetics, supplements and as a health food.

  19. Synthesis of stable isotope enriched metabolites of theophylline

    Four isotopically labelled metabolites of theophylline were synthesized. The syntheses of 1-(methyl-d3)uric-2,8-13C 2-acid; 1,3-di(methyl-d3)uric-2,8-13C2 acid; 3-(methyl-d3)xanthine-2-13C,7-15N and 1-(methyl-d3)xanthine-2-13C are described. Modifications in published procedures for the synthesis of the corresponding unlabelled compounds were required to achieve the micro scale necessary to accommodate the small starting quantities of expensive stable isotope labelled precursors. (author)

  20. [Neurochemical study of effects of the new anxiolytic drugs afobazol and ladasten on the synthesis and metabolism of monoamines and their metabolites in the brain structures of Wistar rat on the model of monoamine synthesis blockade induced by aromatic amino acid decarboxylase inhibitor NSD-1015].

    Davydova, A I; Klodt, P M; Kudrin, V S; Kuznetsova, E A; Narkevich, V B

    2010-03-01

    Results of a neurochemical study of the effects of the new anxiolytic drugs afobazole and ladasten on the synthesis and metabolism of monoamines and their metabolites determined by HPLC on the model of monoamine synthesis blockade induced by NSD-1015 (aromatic L-amino acid decarboxylase) in the brain structures of Wistar rats are reported. A decrease in the levels of DOPAC in hypothalamus and HVA in striatum after afobazole injection may be evidence of an inhibitory action of this drug on the activity of monoamine oxidase (MAO-A), which is the main enzyme involved in dopamine biodegradation. Afobazole was also found to increase the content of serotonin (5-HT) as well as its precursor (5-OTP) and its main metabolite (5-HIAA) in hypothalamus by up to 50, 60 and 50%, respectively, which confirms a hypothesis that this anxiolytic drug can modulate the activity of tryptophan hydroxylase (5-OTP synthesis enzyme). In contrast to afobazole, ladasten demonstrated the ability to increase the level of L-DOPA (a dopamine precursor) in virtually all functional structures of the brain (except for hippocamp), which may support the hypothesis suggestion concerning a predominant action of this drug on the activity of tyrosine hydroxylase. Ladasten exhibited selectivity with respect to the dopaminergic system and affected only parameters of the dopamine metabolism, in particular, by increasing the HVA content in nucleus accumbens and decreasing it in the hypothalamus. The drug also affected the dopamine turnover parameters, producing an increase in both HVA/dopamine ratio in nucleus accumbens and DOPAC/dopamine ratio in hippocamp. PMID:20408420

  1. Metabolites identification of bioactive licorice compounds in rats.

    Wang, Qi; Qian, Yi; Wang, Qing; Yang, Yan-Fang; Ji, Shuai; Song, Wei; Qiao, Xue; Guo, De-An; Liang, Hong; Ye, Min

    2015-11-10

    Licorice (Glycyrrhiza uralensis Fisch.) is one of the most popular herbal medicines worldwide. This study aims to identify the metabolites of seven representative bioactive licorice compounds in rats. These compounds include 22β-acetoxyl glycyrrhizin (1), licoflavonol (2), licoricidin (3), licoisoflavanone (4), isoglycycoumarin (5), semilicoisoflavone B (6), and 3-methoxy-9-hydroxy-pterocarpan (7). After oral administration of 250mg/kg of 1 or 40mg/kg of 2-7 to rats, a total of 16, 43 and 31 metabolites were detected in the plasma, urine and fecal samples, respectively. The metabolites were characterized by HPLC/DAD/ESI-MS(n) and LC/IT-TOF-MS analyses. Particularly, two metabolites of 1 were unambiguously identified by comparing with reference standards, and 22β-acetoxyl glycyrrhizin-6″-methyl ester (1-M2) is a new compound. Compound 1 could be readily hydrolyzed to eliminate the glucuronic acid residue. The phenolic compounds (4-7) mainly undertook phase II metabolism (glucuronidation or sulfation). Most phenolic compounds with an isoprenyl group (chain or cyclized, 2-5) could also undertake hydroxylation reaction. This is the first study on in vivo metabolism of these licorice compounds. PMID:26311472

  2. Metabolic Profiling and Antioxidant Assay of Metabolites from Three Radish Cultivars (Raphanus sativus).

    Park, Chang Ha; Baskar, Thanislas Bastin; Park, Soo-Yun; Kim, Sun-Ju; Valan Arasu, Mariadhas; Al-Dhabi, Naif Abdullah; Kim, Jae Kwang; Park, Sang Un

    2016-01-01

    A total of 13 anthocyanins and 33 metabolites; including organic acids, phenolic acids, amino acids, organic compounds, sugar acids, sugar alcohols, and sugars, were profiled in three radish cultivars by using high-performance liquid chromatography (HPLC) and gas chromatography time-of-flight mass spectrometry (GC-TOFMS)-based metabolite profiling. Total phenolics and flavonoids and their in vitro antioxidant activities were assessed. Pelargonidins were found to be the major anthocyanin in the cultivars studied. The cultivar Man Tang Hong showed the highest level of anthocyanins (1.89 ± 0.07 mg/g), phenolics (0.0664 ± 0.0033 mg/g) and flavonoids (0.0096 ± 0.0004 mg/g). Here; the variation of secondary metabolites in the radishes is described, as well as their association with primary metabolites. The low-molecular-weight hydrophilic metabolite profiles were subjected to principal component analysis (PCA), hierarchical clustering analysis (HCA), Pearson's correlation analysis. PCA fully distinguished the three radish cultivars tested. The polar metabolites were strongly correlated between metabolites that participate in the TCA cycle. The chemometrics results revealed that TCA cycle intermediates and free phenolic acids as well as anthocyanins were higher in the cultivar Man Tang Hong than in the others. Furthermore; superoxide radical scavenging activities and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging were investigated to elucidate the antioxidant activity of secondary metabolites in the cultivars. Man Tang Hong showed the highest superoxide radical scavenging activity (68.87%) at 1000 μg/mL, and DPPH activity (20.78%), followed by Seo Ho and then Hong Feng No. 1. The results demonstrate that GC-TOFMS-based metabolite profiling, integrated with chemometrics, is an applicable method for distinguishing phenotypic variation and determining biochemical reactions connecting primary and secondary metabolism. Therefore; this study might provide

  3. Metabolic Profiling and Antioxidant Assay of Metabolites from Three Radish Cultivars (Raphanus sativus

    Chang Ha Park

    2016-01-01

    Full Text Available A total of 13 anthocyanins and 33 metabolites; including organic acids, phenolic acids, amino acids, organic compounds, sugar acids, sugar alcohols, and sugars, were profiled in three radish cultivars by using high-performance liquid chromatography (HPLC and gas chromatography time-of-flight mass spectrometry (GC-TOFMS-based metabolite profiling. Total phenolics and flavonoids and their in vitro antioxidant activities were assessed. Pelargonidins were found to be the major anthocyanin in the cultivars studied. The cultivar Man Tang Hong showed the highest level of anthocyanins (1.89 ± 0.07 mg/g, phenolics (0.0664 ± 0.0033 mg/g and flavonoids (0.0096 ± 0.0004 mg/g. Here; the variation of secondary metabolites in the radishes is described, as well as their association with primary metabolites. The low-molecular-weight hydrophilic metabolite profiles were subjected to principal component analysis (PCA, hierarchical clustering analysis (HCA, Pearson’s correlation analysis. PCA fully distinguished the three radish cultivars tested. The polar metabolites were strongly correlated between metabolites that participate in the TCA cycle. The chemometrics results revealed that TCA cycle intermediates and free phenolic acids as well as anthocyanins were higher in the cultivar Man Tang Hong than in the others. Furthermore; superoxide radical scavenging activities and 1,1-diphenyl-2-picrylhydrazyl (DPPH radical scavenging were investigated to elucidate the antioxidant activity of secondary metabolites in the cultivars. Man Tang Hong showed the highest superoxide radical scavenging activity (68.87% at 1000 μg/mL, and DPPH activity (20.78%, followed by Seo Ho and then Hong Feng No. 1. The results demonstrate that GC-TOFMS-based metabolite profiling, integrated with chemometrics, is an applicable method for distinguishing phenotypic variation and determining biochemical reactions connecting primary and secondary metabolism. Therefore; this study might

  4. Evaluation of Extraction Protocols for Simultaneous Polar and Non-Polar Yeast Metabolite Analysis Using Multivariate Projection Methods

    Nicolas P. Tambellini

    2013-07-01

    Full Text Available Metabolomic and lipidomic approaches aim to measure metabolites or lipids in the cell. Metabolite extraction is a key step in obtaining useful and reliable data for successful metabolite studies. Significant efforts have been made to identify the optimal extraction protocol for various platforms and biological systems, for both polar and non-polar metabolites. Here we report an approach utilizing chemoinformatics for systematic comparison of protocols to extract both from a single sample of the model yeast organism Saccharomyces cerevisiae. Three chloroform/methanol/water partitioning based extraction protocols found in literature were evaluated for their effectiveness at reproducibly extracting both polar and non-polar metabolites. Fatty acid methyl esters and methoxyamine/trimethylsilyl derivatized aqueous compounds were analyzed by gas chromatography mass spectrometry to evaluate non-polar or polar metabolite analysis. The comparative breadth and amount of recovered metabolites was evaluated using multivariate projection methods. This approach identified an optimal protocol consisting of 64 identified polar metabolites from 105 ion hits and 12 fatty acids recovered, and will potentially attenuate the error and variation associated with combining metabolite profiles from different samples for untargeted analysis with both polar and non-polar analytes. It also confirmed the value of using multivariate projection methods to compare established extraction protocols.

  5. Lanthanum rather than cadmium induces oxidative stress and metabolite changes in Hypericum perforatum

    Highlights: • Impact of La, Cd and Cd + La on the metabolism of Hypericum perforatum was compared. • La stimulated ROS and suppressed growth and basic antioxidants more than Cd. • Impact of Cd + La was not synergistic including the sod gene expression. • La depleted hypericin and hyp-1 gene expression but amount of hyperforin increased. • La reduced flavonols and procyanidins mainly in the roots and affected anatomy - Abstract: Physiology, oxidative stress and production of metabolites in Hypericum perforatum exposed to moderate Cd and/or La concentration (10 μM) were studied. La evoked increase in reactive oxygen species, malondialdehyde and proline but suppressed growth, tissue water content, glutathione, ascorbic acid and affected mineral nutrient contents more than Cd while the impact of Cd + La was not synergistic. Similar trend was observed at the level of superoxide dismutase gene expression. Shoot Cd amount increased in Cd + La while only root La increased in the same treatment. Extensive quantification of secondary metabolites revealed that La affected phenolic acids more pronouncedly than Cd in shoots and roots. Flavonols were suppressed by La that could contribute to the appearance of oxidative damage. Procyanidins increased in response to La in the shoots but decreased in the roots. Metabolic responses in Cd + La treatment resembled those of La treatment (almost identically in the roots). Phenylalanine ammonia-lyase activity was mainly suppressed by La. The presence of La also depleted amount of hypericin and expression of its putative gene (hyp-1) showed similar trend but accumulation of hyperforin increased under Cd or La excess. Clear differences in the stem and root anatomy in response to Cd or La were also found. Overall, H. perforatum is La-sensitive species and rather Cd ameliorated negative impact of La

  6. Lanthanum rather than cadmium induces oxidative stress and metabolite changes in Hypericum perforatum

    Babula, Petr [Department of Physiology, Faculty of Medicine, Masaryk University, Kamenice 753/5, 625 00 Brno (Czech Republic); Klejdus, Bořivoj [Institute of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University in Brno, Zemědělská 1, 613 00 Brno (Czech Republic); CEITEC–Central European Institute of Technology, Mendel University in Brno, Zemědělská 1, 613 00 Brno (Czech Republic); Kovacik, Jozef, E-mail: jozkovacik@yahoo.com [Institute of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University in Brno, Zemědělská 1, 613 00 Brno (Czech Republic); CEITEC–Central European Institute of Technology, Mendel University in Brno, Zemědělská 1, 613 00 Brno (Czech Republic); Hedbavny, Josef; Hlavna, Marián [Institute of Chemistry and Biochemistry, Faculty of Agronomy, Mendel University in Brno, Zemědělská 1, 613 00 Brno (Czech Republic)

    2015-04-09

    Highlights: • Impact of La, Cd and Cd + La on the metabolism of Hypericum perforatum was compared. • La stimulated ROS and suppressed growth and basic antioxidants more than Cd. • Impact of Cd + La was not synergistic including the sod gene expression. • La depleted hypericin and hyp-1 gene expression but amount of hyperforin increased. • La reduced flavonols and procyanidins mainly in the roots and affected anatomy - Abstract: Physiology, oxidative stress and production of metabolites in Hypericum perforatum exposed to moderate Cd and/or La concentration (10 μM) were studied. La evoked increase in reactive oxygen species, malondialdehyde and proline but suppressed growth, tissue water content, glutathione, ascorbic acid and affected mineral nutrient contents more than Cd while the impact of Cd + La was not synergistic. Similar trend was observed at the level of superoxide dismutase gene expression. Shoot Cd amount increased in Cd + La while only root La increased in the same treatment. Extensive quantification of secondary metabolites revealed that La affected phenolic acids more pronouncedly than Cd in shoots and roots. Flavonols were suppressed by La that could contribute to the appearance of oxidative damage. Procyanidins increased in response to La in the shoots but decreased in the roots. Metabolic responses in Cd + La treatment resembled those of La treatment (almost identically in the roots). Phenylalanine ammonia-lyase activity was mainly suppressed by La. The presence of La also depleted amount of hypericin and expression of its putative gene (hyp-1) showed similar trend but accumulation of hyperforin increased under Cd or La excess. Clear differences in the stem and root anatomy in response to Cd or La were also found. Overall, H. perforatum is La-sensitive species and rather Cd ameliorated negative impact of La.

  7. INTERCELLULAR AND INTRACELLULAR TRAFFICKING OF NAD+ AND NAD+ PRECURSORS, INTERMEDIATE METABOLITES AND SIGNAL METABOLITES

    Antonio De Flora

    2012-08-01

    Full Text Available Although being currently illustrated in Biochemistry textbooks, NAD+ metabolism is still largely undefined in its features. Specifically, enzymes involved in NAD+ biosynthesis and some of the enzymes involved in its utilization localize to distinct subcellular compartments of the same cell and, functionally, also to different cell types of the same organism. These findings lead to revolutionize current ideas. For instance, 1 NAD+ biosynthesis from several precursors (e.g., Nicotinamide, Nicotinic Acid, Nicotinamide mononucleotide, Nicotinamide riboside, Tryptophan, collectively defined Vit. B3 is a systemic yet segmentary process, whose individual steps may occur in different cells/tissues/organs. These activate a crosstalk via the exchange of intermediate metabolites in biological fluids and the eventual NAD+ biosynthesis takes place in selected cells able to utilize it in diverse, fundamental processes. Therefore, NAD+ metabolism is an organ- ismal process encompassing local events. 2 Utilization of NAD+ for regulation of cell functions involves the trafficking, both subcellular (autocrine and intercellular (paracrine, of signal-metabolites including NAD+ itself and NAD+-derived second messengers, e.g. Cyclic ADP-ribose and ADP-ribose. This hitherto unrecognized trafficking involves a complex in-terplay of ectoenzymes (e.g. CD38, plasmamembrane receptors and related signal transduction pathways, equilibrative and concentrative transporters, ion channels, whose outcome is the fine control of intracellular Ca2+ homeostasis and of Ca2+-dependent cell functions. Further elucidation of compartmentation of NAD+ and more extensive identification of its precursors/metabolites is expected to unveil at the mechanistic level a number of physiological and pathological processes, e.g. aging and age-related diseases.

  8. Cerebrospinal fluid monoamine metabolites and suicide.

    Jokinen, Jussi; Nordström, Anna-Lena; Nordström, Peter

    2009-01-01

    Prospective studies of the serotonergic system and suicide report that low 5-hydroxyindolacetic acid (5-HIAA) in the cerebrospinal fluid (CSF) and a history of attempted suicide predict suicide risk. Low CSF homovanillic acid (HVA) is reported to be associated with past and future lethality of suicide attempts but not with suicide. The interrelationships between monoamine metabolites, violent method, suicide intent and lethality of suicidal behaviour are complex. We hypothesized that CSF 5-HIAA and HVA levels are related to suicide intent, violence and lethality of suicidal behaviour. Fifteen male suicide attempters admitted to a psychiatric ward at the Karolinska University Hospital and eight healthy male volunteers were submitted to lumbar puncture and CSF 5-HIAA and HVA were assayed. Suicide intent with the Beck Suicide Intent Scale (SIS), lethality and violence of suicidal behaviour were assessed. All patients were followed up for causes of death. Six suicides and one fatal accident were identified with death certificates. Mean CSF 5-HIAA but not CSF HVA differed between suicides and survivors. Violent suicides had higher suicide intent and CSF 5-HIAA than non-violent suicides. In violent suicides, CSF 5-HIAA levels were negatively correlated with SIS. Greater suicide intent may be associated with greater aggressive intent and predicts a violent suicide method. PMID:19034712

  9. Antidiabetic potential and secondary metabolites screening of mangrove gastropod Cerithidea obtusa

    Reni Tri Cahyani; Sri Purwaningsih; Azrifitria

    2015-01-01

    Objective: To study the possible effects of Cerithidea obtusa extract as antidiabetic and to screen the secondary metabolites presence. Methods: Antidiabetic activity of Cerithidea obtusa extract was measured in vitro usingα-glucosidase inhibition method. Whereas, secondary metabolites screening was measured qualitatively. Results: The methanol extract had antidiabetic activity (IC50 = 36.40 mg/mL). However, the control drug acarbose had significantly higher antidiabetic activity (IC50 = 0.32 mg/mL). Secondary metabolites screening showed the presence of alkaloids, flavonoids, triterpenoids and saponins. Conclusions: The methanol extract had antidiabetic activity and the presence of alkaloids, flavonoids and triterpenoids might contribute to the activity.

  10. Chemotyping the distribution of vitamin D metabolites in human serum

    Müller, Miriam J.; Stokes, Caroline S.; Lammert, Frank; Volmer, Dietrich A.

    2016-02-01

    Most studies examining the relationships between vitamin D and disease or health focus on the main 25-hydroxyvitamin D3 (25(OH)D3) metabolite, thus potentially overlooking contributions and dynamic effects of other vitamin D metabolites, the crucial roles of several of which have been previously demonstrated. The ideal assay would determine all relevant high and low-abundant vitamin D species simultaneously. We describe a sensitive quantitative assay for determining the chemotypes of vitamin D metabolites from serum after derivatisation and ultra-high performance liquid chromatography-electrospray ionisation-tandem mass spectrometry (UHPLC-ESI-MS/MS). We performed a validation according to the ‘FDA Guidance for Industry Bioanalytical Method Validation’. The proof-of-concept of the method was then demonstrated by following the metabolite concentrations in patients with chronic liver diseases (CLD) during the course of a vitamin D supplementation study. The new quantitative profiling assay provided highly sensitive, precise and accurate chemotypes of the vitamin D metabolic process rather than the usually determined 25(OH)D3 concentrations.

  11. Simultaneous Qualitative Assessment and Quantitative Analysis of Metabolites (Phenolics, Nucleosides and Amino Acids) from the Roots of Fresh Gastrodia elata Using UPLC-ESI-Triple Quadrupole Ion MS and ESI- Linear Ion Trap High-Resolution MS

    Sha Chen; Jun Qiu Liu; Hui Xiao; Jun Zhang; An Liu

    2016-01-01

    A sensitive, effective and optimized method, based on ultra performance liquid chromatography (UPLC) coupled with ESI-triple quadrupole ion MS and ESI-linear ion trap high-resolution MS, has been developed for the simultaneous quantitative and qualitative determination of phenolics, nucleosides and amino acids in the roots of fresh Gastrodia elata. Optimization of the analytical method provided higher separation efficiency and better peak resolution for the targeted compounds. The simultaneou...

  12. Novel rapid liquid chromatography tandem masspectrometry method for vemurafenib and metabolites in human plasma, including metabolite concentrations at steady state.

    Vikingsson, Svante; Strömqvist, Malin; Svedberg, Anna; Hansson, Johan; Höiom, Veronica; Gréen, Henrik

    2016-08-01

    A novel, rapid and sensitive liquid chromatography tandem-mass spectrometry method for quantification of vemurafenib in human plasma, that also for the first time allows for metabolite semi-quantification, was developed and validated to support clinical trials and therapeutic drug monitoring. Vemurafenib was analysed by precipitation with methanol followed by a 1.9 min isocratic liquid chromatography tandem masspectrometry analysis using an Acquity BEH C18 column with methanol and formic acid using isotope labelled internal standards. Analytes were detected in multireaction monitoring mode on a Xevo TQ. Semi-quantification of vemurafenib metabolites was performed using the same analytical system and sample preparation with gradient elution. The vemurafenib method was successfully validated in the range 0.5-100 μg/mL according to international guidelines. The metabolite method was partially validated owing to the lack of commercially available reference materials. For the first time concentration levels at steady state for melanoma patients treated with vemurafenib is presented. The low abundance of vemurafenib metabolites suggests that they lack clinical significance. Copyright © 2016 John Wiley & Sons, Ltd. PMID:26683023

  13. Establishment of a yeast platform strain for production of p-coumaric acid through metabolic engineering of aromatic amino acid biosynthesis

    Rodriguez Prado, Edith Angelica; Kildegaard, Kanchana Rueksomtawin; Li, Mingji;

    2015-01-01

    Aromatic amino acids are precursors of numerous plant secondary metabolites with diverse biological functions. Many of these secondary metabolites are already being used as active pharmaceutical or nutraceutical ingredients, and there are numerous exploratory studies of other compounds with promi...

  14. Activation of Astroglial Calcium Signaling by Endogenous Metabolites Succinate and Gamma-Hydroxybutyrate in the Nucleus Accumbens

    Molnár, Tünde; Héja, László; Emri, Zsuzsa; Simon, Ágnes; Nyitrai, Gabriella; Pál, Ildikó; Kardos, Julianna

    2011-01-01

    Accumulating evidence suggests that different energy metabolites play a role not only in neuronal but also in glial signaling. Recently, astroglial Ca2+ transients evoked by the major citric acid cycle metabolite succinate (SUC) and gamma-hydroxybutyrate (GHB) that enters the citric acid cycle via SUC have been described in the brain reward area, the nucleus accumbens (NAc). Cells responding to SUC by Ca2+ transient constitute a subset of ATP-responsive astrocytes that are activated in a neur...

  15. Activation of astroglial calcium signaling by endogenous metabolites succinate and gamma-hydroxybutyrate in the nucleus accumbens

    Zsuzsa Emri; Julianna Kardos

    2011-01-01

    Accumulating evidence suggests that different energy metabolites play a role not only in neuronal but also in glial signalling. Recently, astroglial Ca2+ transients evoked by the major citric acid cycle metabolite succinate (SUC) and gamma-hydroxybutyrate (GHB) that enters the citric acid cycle via SUC have been described in the brain reward area, the nucleus accumbens (NAc). Cells responding to SUC by Ca2+ transient constitute a subset of ATP-responsive astrocytes that are activated in a neu...

  16. Influence of the RelA activity on E. coli metabolism by metabolite profiling of glucose-limited chemostat cultures

    Sónia Carneiro; Villas-Bôas, Silas G.; Ferreira, Eugénio C.; Isabel Rocha

    2012-01-01

    Metabolite profiling of E. coli W3110 and the isogenic DrelA mutant cells was used to characterize the RelA-dependent stringent control of metabolism under different growth conditions. Metabolic profiles were obtained by gas chromatography–mass spectrometry (GC-MS) analysis and revealed significant differences between E. coli strains grown at different conditions. Major differences between the two strains were assessed in the levels of amino acids and fatty acids and their precursor metabolit...

  17. Low-molecular-weight metabolite systems chemistry

    Franz eHadacek

    2015-03-01

    Full Text Available Low-molecular-weight metabolites (LMWMs comprise primary or central and a plethora of intermediary or secondary metabolites, all of which are characterized by a molecular weight below 900 Dalton. The latter are especially prominent in sessile higher organisms, such as plants, corals, sponges and fungi, but are produced by all types of microbial organisms too. Common to all of these carbon molecules are oxygen, nitrogen and, to a lesser extent, sulfur, as heteroatoms. The latter can contribute as electron donators or acceptors to cellular redox chemistry and define the potential of the molecule to enter charge-transfer complexes. Furthermore, they allow LMWMs to serve as organic ligands in coordination complexes of various inorganic metals as central atoms. Especially the transition metals Fe, Cu and Mn can catalyze one electron reduction of molecular oxygen, which results in formation of free radical species and reactive follow-up reaction products. As antioxidants LMWMs can scavenge free radicals. Depending on the chemical environment, the same LMWMs can act as pro-oxidants by reducing molecular oxygen. The cellular regulation of redox homeostasis, a balance between oxidation and reduction, is still far from being understood. Charge-transfer and coordination complex formation with metals shapes LMWMs into gel-like matrices in the cytosol. The quasi-polymer structure is lost usually during the isolation procedure. In the gel state, LMWMs possess semiconductor properties. Also proteins and membranes are semiconductors. Together they can represent biotransistor components that can be part of a chemoelectrical signaling system that coordinates systems chemistry by initiating cell differentiation or tissue homeostasis, the activated and the resting cell state, when it is required. This concept is not new and dates back to Albert Szent-Györgyi.

  18. Contribution to the study of beryllium complexes. IX. Chelation of Be2+ ions by 3-hydroxy butanoic and 2-hydroxy 2-methyl propanoic acids, potentiometric study

    In the field of work on complexation of beryllium, complex equilibria in aqueous solution between beryllium (II) and 3-hydroxybutanoic acid (HA) and 2-hydroxy-2-methyl propanoic acid (HA) were studied by potentiometric (alkalimetric) titration (at ionic strength = 0.5 M by NaClO4 and a temperature T = 25.0 +- 0.10C to show the influence of hydroxyl group in carboxylic ligands. For the system 3-hydroxy butanoic acid, the results show the presence of two mononuclear species: BeA+ and BeA2 and one hydrolized complex: Be3(OH)3A2, but for 2-hydroxy-2-methylpropanoic acid an additional species is identified, in what the alcoolic function is dissociated: Be(H-1A). The formation constants are reported

  19. Establishing an Ion Pair Interaction in the Homomeric {rho}1 {gamma}-Aminobutyric Acid Type A Receptor That Contributes to the Gating Pathway

    Wang, Jinti; Lester, Henry A.; Dougherty, Dennis A.

    2007-01-01

    {gamma}-Aminobutyric acid type A (GABAA) receptors are members of the Cys-loop superfamily of ligand-gated ion channels. Upon agonist binding, the receptor undergoes a structural transition from the closed to the open state, but the mechanism of gating is not well understood. Here we utilized a combination of conventional mutagenesis and the high precision methodology of unnatural amino acid incorporation to study the gating interface of the human homopentameric {rho}1 GABAA receptor. We have...

  20. Inhibitory effects of omega-3 fatty acids on injury-induced epidermal growth factor receptor transactivation contribute to delayed wound healing

    Turk, Harmony F.; Monk, Jennifer M.; Fan, Yang-Yi; Callaway, Evelyn S.; Weeks, Brad; Chapkin, Robert S.

    2013-01-01

    Epidermal growth factor receptor (EGFR)-mediated signaling is required for optimal intestinal wound healing. Since n-3 polyunsaturated fatty acids (PUFA), specifically docosahexaenoic acid (DHA), alter EGFR signaling and suppress downstream activation of key signaling pathways, we hypothesized that DHA would be detrimental to the process of intestinal wound healing. Using a mouse immortalized colonocyte model, DHA uniquely reduced EGFR ligand-induced receptor activation, whereas DHA and its m...

  1. Contribution of carbon fixed by Rubisco and PEPC to phloem export in the Crassulacean acid metabolism plant Kalanchoë daigremontiana

    Wild, Birgit; Wanek, Wolfgang; Postl, Wolfgang; Richter, Andreas

    2010-01-01

    Crassulacean acid metabolism (CAM) plants exhibit a complex interplay between CO2 fixation by phosphoenolpyruvate carboxylase (PEPC) and ribulose-1,5-bisphosphate carboxylase oxygenase (Rubisco), and carbon demand for CAM maintenance and growth. This study investigated the flux of carbon from PEPC and direct Rubisco fixation to different leaf carbon pools and to phloem sap over the diurnal cycle. Concentrations and carbon isotope compositions of starch, soluble sugars, and organic acids were ...

  2. The contribution of phenolic acids to the anti-inflammatory activity of mushrooms: screening in phenolic extracts, individual parent molecules and synthesized glucuronated and methylated derivatives

    Ayodele, Oludemi Taofiq; Calhelha, Ricardo C.; Heleno, Sandrina A.; Barros, Lillian; Martins, Anabela; Santos-Buelga, Celestino; Queiroz, Maria João R. P.; Isabel C. F. R. Ferreira

    2015-01-01

    In the present study, the ethanolic extracts of fourteen edible mushrooms were investigated for their anti-inflammatory potential in LPS (lipopolysaccharide) activated RAW 264.7 macrophages. Furthermore the extracts were chemically characterized in terms of phenolic acids and related compounds. The identified molecules (p-hydroxybenzoic, p-coumaric and cinnamic acids) and their glucuronated and methylated derivatives obtained by chemical synthesis were also evaluated for the same bioactivity,...

  3. Polyunsaturated fatty acids and inflammation

    Calder Philip C

    2004-01-01

    The n-6 polyunsaturated fatty acid arachidonic acid gives rise to the eicosanoid family of inflammatory mediators (prostaglandins, leukotrienes and related metabolites) and through these regulates the activities of inflammatory cells, the production of cytokines and the various balances within the immune system. Fish oil and oily fish are good sources of long chain n-3 polyunsaturated fatty acids. Consumption of these fatty acids decreases the amount of arachidonic acid in cell membranes and ...

  4. 气相色谱-火焰离子化检测器检测血清中二醇及其代谢产物羟乙酸%Determination of ethylene glycol and its metabolite glycolic acid in human serum using gas chromatography-flame ionization detector

    于天晓; 李青; 万涛; 丁世家

    2011-01-01

    A gas chromatography method with flame ionization detector for the determination of ethylene glycol (EG) and its toxic metabolite glycolic acid (GA) in serum was established. The sample preparation and derivatization methods were optimized. The EG and its metabolite GA in standard fluid and mouse model serum were determined. Good linearities were obtained at the concentration of 0.01 ~ 10μg/mL for EG and 0. 01~5μg/mL for GA with R=0. 9999 and R=0. 9988, respectively. The recoveries were 91% ~106%and 89%~95% and detection limits were 0. 005μg/mL and 0.01μg/mL, for EG and GA, respectively.The method is sensitive, accurate and easy-to-operate, and can be applied to the determination of EG and GA for medicolegal identification and clinical testing.%建立了气相色谱-火焰离子化检测器(FID)检测血清中乙二醇(ethylene glycol,EG)及其代谢产物羟乙酸(glycolic acid,GA)的方法.通过优化样本预处理和衍生化方法,用气相色谱-火焰离子化检测器检测标准工作液和小鼠中毒模型血清中乙二醇及其代谢产物羟乙酸.结果表明,乙二醇与羟乙酸浓度分别在0.01~10μg/mL,0.01~5μg/mL范围内线性良好,其标准曲线的相关系数分别为0.9999与0.9988,相对标准偏差均小于5%,绝对回收率分别为91~106%和89~95%,最低检出限分别为0.005μg/mL和0.01μg/mL.本方法灵敏度高,简单快速,结果准确可靠,适用于法医学鉴定和临床检验中乙二醇与羟乙酸的检测.

  5. Identification of a Classical Mutant in the Industrial Host Aspergillus niger by Systems Genetics: LaeA Is Required for Citric Acid Production and Regulates the Formation of Some Secondary Metabolites

    Niu, Jing; Arentshorst, Mark; Nair, P. Deepa S.;

    2015-01-01

    The asexual filamentous fungus Aspergillus niger is an important industrial cell factory for citric acid production. In this study, we genetically characterized a UV-generated A. niger mutant that was originally isolated as a nonacidifying mutant, which is a desirable trait for industrial enzyme...... with high-throughput genome sequencing to identify the mutation responsible for the nonacidifying phenotype. Since A. niger has no sexual cycle, parasexual genetics was used to generate haploid segregants derived from diploids by loss of whole chromosomes. We found that the nonacidifying phenotype was...

  6. Rumen-protected conjugated linoleic acid supplementation to dairy cows in late pregnancy and early lactation: effects on milk composition, milk yield, blood metabolites and gene expression in liver

    Meyer Heinrich HD

    2010-02-01

    Full Text Available Abstract Background Conjugated linoleic acid (CLA is a collective term for isomers of octadecadienoic acid with conjugated double-bond system. Thus, it was the objective to investigate whether milk composition and metabolic key parameters are affected by adding CLA to the diet of dairy cows in the first four weeks of lactation. Methods A study was carried out with five primiparous cows fed a CLA supplemented diet compared to five primiparous cows without CLA supplementation. CLA supplemented cows received 7.5 g CLA/day (i.e. 50% cis(c9,trans(t11- and 50% t10,c12-CLA starting two weeks before expected calving and 20 g CLA/day (i.e. 50% c9,t11- and 50% t10,c12-CLA throughout day 1 to 28 of lactation. Results The CLA supplement was insufficiently accepted by the animals: only 61.5% of the intended amount was ingested. Fed CLA were detectable in milk fat, whereas contents of c9,t11-CLA and t10,c12-CLA in milk fat were higher for CLA supplemented cows compared to the control group. On average over the entire treatment period, there was a decrease of saturated fatty acids (FA in milk fat of CLA supplemented cows, combined with a higher content of monounsaturated and trans FA. Our study revealed no significant effects of c9,t11- and t10,c12-CLA supplementation either on milk yield and composition or on metabolic key parameters in blood. Furthermore the experiment did not indicate significant effects of c9,t11- and t10,c12-CLA-supplementation on gene expression of peroxisome proliferator-activated receptor-alpha (PPARα, PPARγ, sterol regulatory element-binding protein-1 and tumor necrosis factor-alpha in liver tissue. Conclusions Feeding c9,t11- and t10,c12-CLA during the first weeks after calving did not affect metabolic key parameters of blood serum or milk composition of fresh cows. Milk fatty acid composition was changed by feeding c9,t11- and t10,c12-CLA resulting in higher contents of these isomers in milk fat. High contents of long chain FA in

  7. Alterations of urinary metabolite profile in model diabetic nephropathy

    Stec, Donald F. [Vanderbilt Institute of Chemical Biology, Vanderbilt University Medical Center, Nashville, TN 37232 (United States); Wang, Suwan; Stothers, Cody [Department of Medicine, Vanderbilt University Medical Center, Nashville, TN 37232 (United States); Avance, Josh [Berea College, 1916 CPO, Berea, KY 40404 (United States); Denson, Deon [Choctaw Central High School, Philadelphia, MS 39350 (United States); Harris, Raymond [Department of Medicine, Vanderbilt University Medical Center, Nashville, TN 37232 (United States); Voziyan, Paul, E-mail: paul.voziyan@vanderbilt.edu [Department of Medicine, Vanderbilt University Medical Center, Nashville, TN 37232 (United States)

    2015-01-09

    Highlights: • {sup 1}H NMR spectroscopy was employed to study urinary metabolite profile in diabetic mouse models. • Mouse urinary metabolome showed major changes that are also found in human diabetic nephropathy. • These models can be new tools to study urinary biomarkers that are relevant to human disease. - Abstract: Countering the diabetes pandemic and consequent complications, such as nephropathy, will require better understanding of disease mechanisms and development of new diagnostic methods. Animal models can be versatile tools in studies of diabetic renal disease when model pathology is relevant to human diabetic nephropathy (DN). Diabetic models using endothelial nitric oxide synthase (eNOS) knock-out mice develop major renal lesions characteristic of human disease. However, it is unknown whether they can also reproduce changes in urinary metabolites found in human DN. We employed Type 1 and Type 2 diabetic mouse models of DN, i.e. STZ-eNOS{sup −/−} C57BLKS and eNOS{sup −/−} C57BLKS db/db, with the goal of determining changes in urinary metabolite profile using proton nuclear magnetic resonance (NMR). Six urinary metabolites with significantly lower levels in diabetic compared to control mice have been identified. Specifically, major changes were found in metabolites from tricarboxylic acid (TCA) cycle and aromatic amino acid catabolism including 3-indoxyl sulfate, cis-aconitate, 2-oxoisocaproate, N-phenyl-acetylglycine, 4-hydroxyphenyl acetate, and hippurate. Levels of 4-hydroxyphenyl acetic acid and hippuric acid showed the strongest reverse correlation to albumin-to-creatinine ratio (ACR), which is an indicator of renal damage. Importantly, similar changes in urinary hydroxyphenyl acetate and hippurate were previously reported in human renal disease. We demonstrated that STZ-eNOS{sup −/−} C57BLKS and eNOS{sup −/−} C57BLKS db/db mouse models can recapitulate changes in urinary metabolome found in human DN and therefore can be

  8. Characterization of furanocoumarin metabolites in parsnip webworm, Depressaria pastinacella.

    Nitao, James K; Berhow, Mark; Duval, Sandra M; Weisleder, David; Vaughn, Steven F; Zangerl, Arthur; Berenbaum, M R

    2003-03-01

    Although metabolites of furanocoumarins have been characterized in a wide range of organisms, to date they have been identified in only a single insect species, Papilio polyxenes. Depressaria pastinacella, the parsnip webworm, like P. polyxenes a specialist on Apiaceae, routinely consumes plant tissues higher in furanocoumarin content than does P. polyxenes and is capable of faster cytochrome P-450-mediated detoxification of these compounds. In this study, we characterized metabolites of xanthotoxin, a linear furanocoumarin, and sphondin, an angular furanocoumarin, in midguts and frass of parsnip webworms. Two metabolites were isolated and identified from webworms fed artificial diet containing xanthotoxin. LC-ESI-MS analysis resulted in the determination of a MW of 266 for the compound in the frass and one of the compounds in the midgut; 1H NMR confirmed its structure as 6-(7-hydroxy-8-methoxycoumaryl)-hydroxyacetic acid (HCHA). The second compound from the midgut had a MW of 252 and was identified by 1H NMR and 13C NMR analysis as 6-(7-hydroxy-8-methoxycoumaryl)-hydroxyethanol) (HMCH). Whereas HCHA has been found in frass of Papilio polyxenes fed xanthotoxin, HMCH has not been reported previously in insects. Although the first step of metabolism of xanthotoxin in webworms as well as P. polyxenes is likely the formation of an epoxide on the furan ring, angular furanocoumarin metabolism in webworms appears to differ. The principal metabolite of sphondin was identified as demethylated sphondin (6-hydroxy-2H-furo[2,3-h]-1-benzopyran-2-one) by LC-ESI-MS and confirmed by 1H NMR and 13C NMR analyses. That webworms produce metabolites of xanthotoxin in common not only with other Lepidoptera (e.g., HCHA) but with other vertebrates (e.g., HMCH) suggests a remarkable conservatism in the metabolic capabilities of cytochrome P-450s and raises the possibility that insects may share other detoxification reactions with vertebrates with respect to toxins in foodplants. PMID

  9. DETERMINATION OF PRIMARY METABOLITES IN CEREAL MILLING FRACTIONS

    Eva Ivanišová

    2013-10-01

    Full Text Available The aim of this study was to determine of primary metabolites (content of starch, total dietary fibre, reducing sugars, proteins and amino acids in four milling fractions of selected cereals (barley, wheat, oat, spelt, rye, triticale grew in the year 2009. It was found that flour fractions (break flour and reduction flour showed the lower content of primary metabolites than bran fractions (fine bran and coarse bran. The aim of this study was also to mention the potential use of bran parts of grain - substances from these parts can be isolated and after treatment, which causes their efficiently usable for human body, they can be used for fortification of wide range of food products.

  10. AGE metabolites: a biomarker linked to cancer disparity?

    Foster, Dion; Spruill, Laura; Walter, Katherine R; Nogueira, Lourdes M; Fedarovich, Hleb; Turner, Ryan Y; Ahmed, Mahtabuddin; Salley, Judith D; Ford, Marvella E; Findlay, Victoria J; Turner, David P

    2014-10-01

    Socioeconomic and environmental influences are established factors promoting cancer disparity, but the contribution of biologic factors is not clear. We report a mechanistic link between carbohydrate-derived metabolites and cancer that may provide a biologic consequence of established factors of cancer disparity. Glycation is the nonenzymatic glycosylation of carbohydrates to macromolecules, which produces reactive metabolites called advanced glycation end products (AGE). A sedentary lifestyle and poor diet all promote disease and the AGE accumulation pool in our bodies and also increase cancer risk. We examined AGE metabolites in clinical specimens of African American and European American patients with prostate cancer and found a higher AGE concentration in these specimens among African American patients when compared with European American patients. Elevated AGE levels corresponded with expression of the receptor for AGE (RAGE or AGER). We show that AGE-mediated increases in cancer-associated processes are dependent upon RAGE. Aberrant AGE accumulation may represent a metabolic susceptibility difference that contributes to cancer disparity. PMID:25053712

  11. A convenient method for simultaneous quantification of multiple phytohormones and metabolites: application in study of rice-bacterium interaction

    Liu Hongbo; Li Xianghua; Xiao Jinghua; Wang Shiping

    2012-01-01

    Abstract Background Simultaneous analysis of multiple functional-related phytohormones and their metabolites will improve our understanding of interactions among different hormones in the same biologic process. Results A method was developed for simultaneous quantification of multiple phytohormones, abscisic acid, indole-3-acetic acid (IAA), jasmonic acid (JA), and salicylic acid, hormone conjugates, IAA-aspartic acid, JA-isoleucine, and methyl JA, and phytoalexins, momilactone A, naringenin,...

  12. Antifungal Activity of Microbial Secondary Metabolites

    Jeffrey J Coleman; Ghosh, Suman; Okoli, Ikechukwu; Mylonakis, Eleftherios

    2011-01-01

    Secondary metabolites are well known for their ability to impede other microorganisms. Reanalysis of a screen of natural products using the Caenorhabditis elegans-Candida albicans infection model identified twelve microbial secondary metabolites capable of conferring an increase in survival to infected nematodes. In this screen, the two compound treatments conferring the highest survival rates were members of the epipolythiodioxopiperazine (ETP) family of fungal secondary metabolites, acetylg...

  13. Secondary metabolites in fungus-plant interactions

    Pusztahelyi, Tünde; Holb, Imre J.; Pócsi, István (1961-) (biológus)

    2015-01-01

    Fungi and plants are rich sources of thousands of secondary metabolites. The genetically coded possibilities for secondary metabolite production, the stimuli of the production, and the special phytotoxins basically determine the microscopic fungi-host plant interactions and the pathogenic lifestyle of fungi. The review introduces plant secondary metabolites usually with antifungal effect as well as the importance of signaling molecules in induced systemic resistance and systemic acquired resi...

  14. Probabilistic Approach for Evaluating Metabolite Sample Integrity

    Slaff, Barry M.; Jensen, Shane T.; Weljie, Aalim M.

    2015-01-01

    The success of metabolomics studies depends upon the "fitness" of each biological sample used for analysis: it is critical that metabolite levels reported for a biological sample represent an accurate snapshot of the studied organism's metabolite profile at time of sample collection. Numerous factors may compromise metabolite sample fitness, including chemical and biological factors which intervene during sample collection, handling, storage, and preparation for analysis. We propose a probabi...

  15. Biochemical parameters as biomarkers for the early recognition of environmental pollution on Scots pine trees. II. The antioxidative metabolites ascorbic acid, glutathione, {alpha}-tocopherol and the enzymes superoxide dismutase and glutathione reductase

    Schulz, H.; Haertling, S. [UFZ Centre for Environmental Research Leipzig-Halle, Halle (Germany). Dept. of Soil Sciences

    2001-10-01

    Field investigations with Scots pine trees (Pinus sylvestris L.) were performed in eastern Germany, where ambient SO{sub 2}, NO{sub x} and O{sub 3} concentrations differed significantly in 1992-99 at three sites, namely Neuglobsow (yearly mean SO{sub 2} in 1992: 9 {mu}g m{sup -3}), Taura (yearly mean SO{sub 2} in 1992: 54 {mu}g m{sup -3}) and Roesa (yearly mean SO{sub 2} in 1992: 73 {mu}g m{sup -3}). To investigate the effects of SO{sub 2}, NO{sub x} and O{sub 3} on antioxidants (superoxide dismutase, ascorbic acid, glutathione, glutathione reductase, {alpha}-tocopherol) and pigments including chlorophyll fluorescence as well as visible damage symptoms in the form of needle yellowing and tip necroses, needles of the 1st and 2nd age class from young and mature trees were collected at the sites every October. Eight years after the start of the field study in 1992, the ambient SO{sub 2} concentrations had decreased significantly at Neuglobsow (yearly mean SO{sub 2} in 1999: 4 {mu}g m{sup -3}), Taura (yearly mean SO{sub 2} in 1999: 5 {mu}g m{sup -3}) and Roesa (yearly mean SO{sub 2} in 1999: 5 {mu}g m{sup -3}). NO{sub x} and O{sub 3} differed less at the three sites and showed no temporal variations. Whole needle glutathione continuously decreased, although concentrations were higher in needles of the 1st and 2nd age class from the polluted sites Taura and Roesa than the unpolluted site Neuglobsow. The activities of glutathione reductase exhibited the same site-related differences and temporal variations and were correlated with concentrations of oxidized glutathione (GSSG). In contrast, the activities of the enzyme superoxide dismutase and the concentrations of whole needle ascorbic acid remained unchanged over the period. Only at the end of the investigation period did the concentrations of oxidized ascorbic acid (dehydroascorbate) increase in six-month-old needles at the polluted sites Taura and Roesa. Despite the clear decreases in SO{sub 2}, the visible symptoms

  16. Isolation and identification of metabolites of porfiromycin formed in the presence of a rat liver preparation.

    Lang, W; Mao, J; Wang, Q; Niu, C; Doyle, T W; Almassian, B

    2000-02-01

    The isolation and identification of the major metabolites of porfiromycin formed in the presence of a rat liver preparation under aerobic conditions were performed with high-performance liquid chromatography and electrospray ionization mass spectrometry. Porfiromycin was extensively metabolized by the rat liver preparation in an aqueous 0.1 M potassium phosphate buffer (pH 7.4) containing an NADPH generating system at 37 degrees C. A total of eight metabolites was identified as mitosene analogs. Of these, three primary metabolites are 2-methylamino-7-aminomitosene, 1,2-cis and 1,2-trans-1-hydroxy-2-methylamino-7-aminomitosene, which are consistent with those previously observed in hypoxia using purified rat liver NADPH-cytochrome c reductase. Interestingly, 2-methylamino-7-aminomitosene is a reactive metabolite, which undergoes further activation at the C-10 position by the loss of carbamic acid and then links with the 7-amino group of the primary metabolites to yield two dimeric adducts. In addition, three phosphate adducts, 10-decarbamoyl-2-methylamino-7-aminomitosene-10-phosphate, 1,2-cis and 1,2-trans-2-methylamino-7-aminomitosene-1-phosphate, were also identified in the incubation system. The configurations of the diastereoisomeric metabolites were determined with (1)HNMR and phosphatase digestion. On the basis of the metabolite profile, we propose in vitro metabolic pathways for porfiromycin. The findings provide direct evidence for understanding the reactive nature and hepatic metabolism of the drug currently in phase III clinical trials. PMID:10688748

  17. Metabolites from Alternaria Fungi and Their Bioactivities

    Ligang Zhou

    2013-05-01

    Full Text Available Alternaria is a cosmopolitan fungal genus widely distributing in soil and organic matter. It includes saprophytic, endophytic and pathogenic species. At least 268 metabolites from Alternaria fungi have been reported in the past few decades. They mainly include nitrogen-containing metabolites, steroids, terpenoids, pyranones, quinones, and phenolics. This review aims to briefly summarize the structurally different metabolites produced by Alternaria fungi, as well as their occurrences, biological activities and functions. Some considerations related to synthesis, biosynthesis, production and applications of the metabolites from Alternaria fungi are also discussed.

  18. Untargeted MS-based small metabolite identification from the plant leaves and stems of Impatiens balsamina.

    Chua, Lee Suan

    2016-09-01

    The identification of plant metabolites is very important for the understanding of plant physiology including plant growth, development and defense mechanism, particularly for herbal medicinal plants. The metabolite profile could possibly be used for future drug discovery since the pharmacological activities of the indigenous herbs have been proven for centuries. An untargeted mass spectrometric approach was used to identify metabolites from the leaves and stems of Impatiens balsamina using LC-DAD-MS/MS. The putative compounds are mostly from the groups of phenolic, organic and amino acids which are essential for plant growth and as intermediates for other compounds. Alanine appeared to be the main amino acid in the plant because many alanine derived metabolites were detected. There are also several secondary metabolites from the groups of benzopyrones, benzofuranones, naphthoquinones, alkaloids and flavonoids. The widely reported bioactive components such as kaempferol, quercetin and their glycosylated, lawsone and its derivatives were detected in this study. The results also revealed that aqueous methanol could extract flavonoids better than water, and mostly, flavonoids were detected from the leaf samples. The score plots of component analysis show that there is a minor variance in the metabolite profiles of water and aqueous methanolic extracts with 21.5 and 30.5% of the total variance for the first principal component at the positive and negative ion modes, respectively. PMID:27135814

  19. Study on the radiation-induced biological responses based on the analysis of metabolites

    1. Objectives □ Establishment of basis of biological radiation response study by metabolite analysis 2. Project results □ Establishment of analytical basis of radiation-responsive metabolites in biological samples - Large scale collection of tissue samples from irradiated animal for radiation metabolomics research - Establishment of mass spectromety (GC MS, LC MS-MS) analysis methods of biological samples - 3 Standard Operation Protocols (SOP) for ultra high resolution mass spectrometry (FT-ICR MS, Q-TOF MS) analysis of metabolites from biological samples - Establishment of database for radiation metabolites □ Basic research on radiation-responsive metabolites and the interpretation of their functions - Validation of spermidine as a candidate biomarker of acute radiation response in mouse blood - Verification of 5 radiation-responsive steroid hormones and alteration of their metabolic enzyme activities in mouse blood - Verification of 13 radiation-responsive amino acids (related to oxidative stress, neurotransmission, energy metabolism) in regional mouse brain -Verification of 10 radiation-responsive amino acids (related to oxidative stress, neurotransmission, energy metabolism) in regional mouse brain - Verification of 74 radiation-responsive metabolites in whole rat brain by ultra high resolution FT-ICR MS and Q-TOF MS analysis 3. Expected benefits and plan of application □ Establishment of research basis of radiation metabolomics in Korea □ Provision of core technology in radiation bioscience and safety field by application of radiation metabolomics results to the technology development in radiation biodosimetry, and radiation response evaluation and modulation

  20. Metabolite Profiling of Root Exudates of Common Bean under Phosphorus Deficiency

    Keitaro Tawaraya

    2014-07-01

    Full Text Available Root exudates improve the nutrient acquisition of plants and affect rhizosphere microbial communities. The plant nutrient status affects the composition of root exudates. The purpose of this study was to examine common bean (Phaseolus vulgaris L. root exudates under phosphorus (P deficiency using a metabolite profiling technique. Common bean plants were grown in a culture solution at P concentrations of 0 (P0, 1 (P1 and 8 (P8 mg P L−1 for 1, 10 and 20 days after transplanting (DAT. Root exudates were collected, and their metabolites were determined by capillary electrophoresis time-of-flight mass spectrometry (CE-TOF MS. The shoot P concentration and dry weight of common bean plants grown at P0 were lower than those grown at P8. One hundred and fifty-nine, 203 and 212 metabolites were identified in the root exudates, and 16% (26/159, 13% (26/203 and 9% (20/212 of metabolites showed a P0/P8 ratio higher than 2.0 at 1, 10 and 20 DAT, respectively. The relative peak areas of several metabolites, including organic acids and amino acids, in root exudates were higher at P0 than at P8. These results suggest that more than 10% of primary and secondary metabolites are induced to exude from roots of common bean by P deficiency.