Sample records for acetoacetates

  1. Evidence for the existence of mammalian acetoacetate decarboxylase: with special reference to human blood serum

    Stekelenburg, Gerard J. van; Koorevaar, Gerrit


    In this article evidence is presented for the existence of mammalian acetoacetate decarboxylase (acetoacetate carboxy-lyase: E.G. From experiments with human blood serum the presence of a non-ultrafiltrable activator, accelerating the decomposition of acetoacetate into acetone and carbon d

  2. Deuterium Exchange in Ethyl Acetoacetate: An Undergraduate GC-MS [Gas Chromatography-Mass Spectroscopy] Experiment

    Heinson, C. D.; Williams, J. M.; Tinnerman, W. N.; Malloy, T. B.


    The role of ethanol O-d in nullifying the deuterolysis may be demonstrated by determining that transesterification of methyl acetoacetate of the ethyl ester occurs as well as deuterium exchange of the five acetoacetate hydrogens. The significant acidity of the methylene protons in the acetoacetate group, the efficacy of base catalysis, the role of…

  3. Whole cells in enantioselective reduction of benzyl acetoacetate

    Joyce Benzaquem Ribeiro


    Full Text Available The β-ketoester benzyl acetoacetate was enantioselectively reduced to benzyl (S-3-hydroxybutanoate by seven microorganism species. The best result using free cells was obtained with the yeast Hansenula sp., which furnished 97% ee and 85% of conversion within 24 h. After immobilization in calcium alginate spheres, K.marxianus showed to be more stable after 2 cycles of reaction.

  4. Stereoselective synthesis of acetoacetate-derived enol triflates.

    Babinski, David; Soltani, Omid; Frantz, Doug E


    A highly stereoselective method for preparing ( Z)- and ( E)-enol triflates derived from substituted acetoacetate derivatives is described. The salient feature of this methodology is the use of Schotten-Baumann-type conditions to control enolate geometry using either aqueous LiOH ( Z-selective) or aqueous (Me)(4)NOH ( E-selective) in combination with triflic anhydride to provide a practical and predictable approach to these valuable substrates.

  5. Acetone production in solventogenic Clostridium species: new insights from non-enzymatic decarboxylation of acetoacetate.

    Han, Bei; Gopalan, Venkat; Ezeji, Thaddeus Chukwuemeka


    Development of a butanologenic strain with high selectivity for butanol production is often proposed as a possible route for improving the economics of biobutanol production by solventogenic Clostridium species. The acetoacetate decarboxylase (aadc) gene encoding acetoacetate decarboxylase (AADC), which catalyzes the decarboxylation of acetoacetate into acetone and CO(2), was successfully disrupted by homologous recombination in solventogenic Clostridium beijerinckii NCIMB 8052 to generate an aadc ( - ) mutant. Our fermentation studies revealed that this mutant produces a maximum acetone concentration of 3 g/L (in P2 medium), a value comparable to that produced by wild-type C. beijerinckii 8052. Therefore, we postulated that AADC-catalyzed decarboxylation of acetoacetate is not the sole means for acetone generation. Our subsequent finding that non-enzymatic decarboxylation of acetoacetate in vitro, under conditions similar to in vivo acetone-butanol-ethanol (ABE) fermentation, produces 1.3 to 5.2 g/L acetone between pH 6.5 and 4 helps rationalize why various knock-out and knock-down strategies designed to disrupt aadc in solventogenic Clostridium species did not eliminate acetone production during ABE fermentation. Based on these results, we discuss alternatives to enhance selectivity for butanol production.

  6. Cosolvent Effect on the Tautomerism of Ethyl Acetoacetate in Supercritical CO2


    The effect of cosolvent cyclohexane, chloroform, and acetonitrile on the keto-enol tautomeric equilibrium of ethyl acetoacetate in supercritical CO2 was studied by UV-Vis spectroscopy over the pressure range from 76 to 110 bar at 308.15 K. It was found that the equilibrium constant decreases with increasing polarity of the cosolvents.

  7. Enhanced and suppressed mineralization by acetoacetate and β-hydroxybutyrate in osteoblast cultures.

    Saito, Akihiro; Yoshimura, Kentaro; Miyamoto, Yoichi; Kaneko, Kotaro; Chikazu, Daichi; Yamamoto, Matsuo; Kamijo, Ryutaro


    It is known that diabetes aggravates alveolar bone loss associated with periodontitis. While insulin depletion increases the blood concentration of ketone bodies, i.e., acetoacetate and β-hydroxybutyrate, their roles in bone metabolism have not been much studied until today. We investigated the effects of acetoacetate and β-hydroxybutyrate on mineralization of extracellular matrix in cultures of mouse osteoblastic MC3T3-E1 cells and primary mouse osteoblasts in the presence and absence of bone morphogenetic protein-2. Acetoacetate potentiated alkaline phosphatase activity in MC3T3-E1 cells in a concentration-dependent manner, ranging from physiological to pathological concentrations (0.05-5 mmol/L). In contrast, β-hydroxybutyrate lowered it in the same experimental settings. Mineralization in cultures of these cells was also up-regulated by acetoacetate and down-regulated by β-hydroxybutyrate. Similar results were obtained in cultures of mouse primary osteoblasts. Neither alkaline phosphatase mRNA nor its protein expression in MC3T3-E1 cells was affected by acetoacetate or β-hydroxybutyrate, indicating that these ketone bodies control the enzyme activity of alkaline phosphatase in osteoblasts and hence their mineralization bi-directionally. Finally, either gene silencing of monocarboxylate transporter-1, a major transmembrate transporter for ketone bodies, nullified the effects of ketone bodies on alkaline phosphatase activity in MC3T3-E1 cells. Collectively, we found that ketone bodies bidirectionally modulates osteoblast functions, which suggests that ketone bodies are important endogenous factors that regulate bone metabolism in both physiological and pathological situations.

  8. Baker's yeast catalyzed asymmetric reduction of methyl acetoacetate in glycerol containing systems

    Adi Wolfson


    Full Text Available The asymmetric hydrogenation of methyl acetoacetate was successfully performed with baker's yeast in pure glycerol and mixtures of glycerol and water. Though yeast viability was very low after exposure to glycerol, the enzymatic activity in pure glycerol was preserved for some days. In addition, a mixture of glycerol and water combined the advantageous of each individual solvent and resulted in high catalytic performance and efficient product extraction yield

  9. Ethyl acetoacetate ligand distribution in the course of titanium n-butoxide chelation

    Kurajica, S., E-mail:; Škorić, I.; Lozić, I.; Mandić, V.


    Sols obtained by chelation of titanium n-butoxide with ethyl acetoacetate, Eaa, in various ratios have been subjected to FTIR, {sup 1}H and {sup 13}C NMR, HSQC and UV–Vis spectroscopy in order to provide insight in the compounds obtained, their structure and quantitative relationships. Three compounds, the bis-chelated monomer, Ti(O{sup n}Bu){sub 2}(Eaa){sub 2}, bis-chelated dimer, (Ti(O{sup n}Bu){sub 3}Eaa){sub 2} and monochelated dimer, Ti{sub 2}(O{sup n}Bu){sub 7}Eaa have been established. As the molar ratio Eaa/Ti(O{sup n}Bu){sub 4} increases, the coordination changes from the monochelated and bis-chelated dimer to the bis-chelated monomer. Additionally, the transesterification reaction, influencing the chemical composition of the compounds was noted. The hydrolysis of the prepared sols was partial, leaving some residual butoxy and ethyl acetoacetate groups attached to titanium. Thermal treatment of the prepared amorphous gels at 350 °C yielded with the formation of nanocrystalline anatase. It was noted that high Eaa/Tnb ratio slightly retards the anatase formation. - Highlights: • Titanium n-butoxide was modified with ethyl acetoacetate in various ratios. • Among other chelate phases, previously undescribed Ti{sub 2}(O{sup n}Bu){sub 7}Eaa was obtained. • NMR-based mass balance procedure was introduced to obtain quantitative relationships. • The transesterification reaction has been noted. • Nanocrystalline anatase has been obtained by thermal treatment at 350 °C.

  10. Automated synthesis of {sup 11}C-acetoacetic acid, a key alternate brain fuel to glucose

    Tremblay, Sebastien [Research Center on Aging, Sherbrooke University Geriatric Institute, 1036 Belvedere Street South, Sherbrooke, Quebec, J1H 4C4 (Canada)]. E-mail:; Ouellet, Rene [Sherbrooke Molecular Imaging Centre, Etienne-LeBel Clinical Research Centre, Centre Hospitalier Universitaire de Sherbrooke, Quebec, J1H 5N4 (Canada); Rodrigue, Serge [Sherbrooke Molecular Imaging Centre, Etienne-LeBel Clinical Research Centre, Centre Hospitalier Universitaire de Sherbrooke, Quebec, J1H 5N4 (Canada); Langlois, Rejean [Sherbrooke Molecular Imaging Centre, Etienne-LeBel Clinical Research Centre, Centre Hospitalier Universitaire de Sherbrooke, Quebec, J1H 5N4 (Canada); Department of Nuclear Medicine and Radiobiology, Quebec, J1H 5N4 (Canada); Benard, Francois [Sherbrooke Molecular Imaging Centre, Etienne-LeBel Clinical Research Centre, Centre Hospitalier Universitaire de Sherbrooke, Quebec, J1H 5N4 (Canada); Department of Nuclear Medicine and Radiobiology, Quebec, J1H 5N4 (Canada); Cunnane, Stephen C. [Research Center on Aging, Sherbrooke University Geriatric Institute, 1036 Belvedere Street South, Sherbrooke, Quebec, J1H 4C4 (Canada); Department of Medicine, Faculty of Medicine and Health Sciences, Universite de Sherbrooke, Sherbrooke, Quebec, J1H 5N4 (Canada)


    An automated, one-pot radio-synthesis module for the routine preparation of 1-[{sup 11}C]acetoacetic acid has been developed. The enolate anion of acetone was reacted with [{sup 11}C]CO{sub 2} in tetrahydrofuran (THF), followed by hydrolysis and purification by ion-exchange chromatography. The total synthesis time was 18 min and radiochemical yield was 34% after decay correction. HPLC analysis showed {<=}3% impurities while residual THF ({<=}200 ppm) and ethanol ({<=}500 ppm) were well under the tolerable limits for human studies.

  11. Acetoacetate promotes the formation of fluorescent advanced glycation end products (AGEs).

    Bohlooli, Mousa; Ghaffari-Moghaddam, Mansour; Khajeh, Mostafa; Aghashiri, Zohre; Sheibani, Nader; Moosavi-Movahedi, Ali Akbar


    Acetoacetate (AA) is an important ketone body, which produces reactive oxygen species (ROS). Advanced glycation end products (AGEs) are defined as final products of glycation process whose production is influenced by the levels of ROS. The accumulation of AGEs in the body contributes to pathogenesis of many diseases including complications of diabetes, and Alzheimer's and Parkinson's disease. Here, we evaluated the impact of AA on production of AGEs upon incubation of human serum albumin (HSA) with glucose. The effect of AA on the AGEs formation of HSA was studied under physiological conditions after incubation with glucose for 35 days. The physical techniques including circular dichroism (CD) and fluorescence spectroscopy were used to assess the impact of AA on formation and structural changes of glycated HSA (GHSA). Our results indicated that the secondary and tertiary structural changes of GHSA were increased in the presence of AA. The fluorescence intensity measurements of AGEs also showed an increase in AGEs formation. Acetoacetate has an activator effect in formation of AGEs through ROS production. The presence of AA may result in enhanced glycation in the presence of glucose and severity of complications associated with accumulation of AGEs.

  12. Physiological and biochemical effects of allelochemical ethyl 2-methyl acetoacetate (EMA) on cyanobacterium Microcystis aeruginosa.

    Hong, Yu; Hu, Hong-Ying; Li, Feng-Min


    The physiological and biochemical effects of an allelochemical ethyl 2-methyl acetoacetate (EMA) isolated from reed (Phragmites communis) on bloom-forming cyanobacterium, Microcystis aeruginosa, were investigated. EMA significantly inhibited the growth of M. aeruginosa in a concentration-dependent way. The metabolic indices (represented by esterase and total dehydrogenase activities), the cellular redox status (represented by the level of reactive oxygen species (ROS)), and the oxidative damage index (represented by the content of malondialdehyde (MDA), the product of membrane lipid peroxidation) were used to evaluate the physiological and biochemical changes in M. aeruginosa after EMA exposure. Esterase activity in M. aeruginosa did not change (P>0.05) after 2 h of exposure to EMA, but increased greatly after 24 and 48 h (PEMA exposure (>0.5 mg L(-1)) resulted in a remarkable loss of total dehydrogenase activity in M. aeruginosa after 4 h (PEMA caused a great increase in ROS level of the algal cells. At high EMA concentration (4 mg L(-1)), the ROS level was remarkably elevated to 1.91 times as much as that in the controls after 2 h. Increases in the ROS level also occurred after 24 and 48 h. The increase in lipid peroxidation of M. aeruginosa was dependent upon EMA concentration and the exposure time. After 40 h of exposure, the MDA content at 4 mg L(-1) of EMA reached approximately 3.5 times (PEMA; the increased metabolic activity perhaps reflects the fact that the resistance of cellular response system to the stress from EMA is initiated during EMA exposure, and the oxidative damage induced by EMA via the oxidation of ROS may be an important factor responsible for the inhibition of EMA on the growth of M. aeruginosa.

  13. Acetoacetate reduces growth and ATP concentration in cancer cell lines which over-express uncoupling protein 2

    Quadros Edward V


    Full Text Available Abstract Background Recent evidence suggests that several human cancers are capable of uncoupling of mitochondrial ATP generation in the presence of intact tricarboxylic acid (TCA enzymes. The goal of the current study was to test the hypothesis that ketone bodies can inhibit cell growth in aggressive cancers and that expression of uncoupling protein 2 is a contributing factor. The proposed mechanism involves inhibition of glycolytic ATP production via a Randle-like cycle while increased uncoupling renders cancers unable to produce compensatory ATP from respiration. Methods Seven aggressive human cancer cell lines, and three control fibroblast lines were grown in vitro in either 10 mM glucose medium (GM, or in glucose plus 10 mM acetoacetate [G+AcA]. The cells were assayed for cell growth, ATP production and expression of UCP2. Results There was a high correlation of cell growth with ATP concentration (r = 0.948 in a continuum across all cell lines. Controls demonstrated normal cell growth and ATP with the lowest density of mitochondrial UCP2 staining while all cancer lines demonstrated proportionally inhibited growth and ATP, and over-expression of UCP2 (p Conclusion Seven human cancer cell lines grown in glucose plus acetoacetate medium showed tightly coupled reduction of growth and ATP concentration. The findings were not observed in control fibroblasts. The observed over-expression of UCP2 in cancer lines, but not in controls, provides a plausible molecular mechanism by which acetoacetate spares normal cells but suppresses growth in cancer lines. The results bear on the hypothesized potential for ketogenic diets as therapeutic strategies.

  14. Chlorine Dioxide-Iodide-Methyl Acetoacetate Oscillation Reaction Investigated by UV-Vis and Online FTIR Spectrophotometric Method

    Shi, Laishun; Wang, Xiaomei; Li, Na; Liu, Jie; Yan, Chunying


    In order to study the chemical oscillatory behavior and mechanism of a new chlorine dioxide-iodide ion-methyl acetoacetate reaction system, a series of experiments were done by using UV-Vis and online FTIR spectrophotometric method. The initial concentrations of methyl acetoacetate, chlorine dioxide, potassium iodide, and sulfuric acid and the pH value have great influence on the oscillation observed at wavelength of 289 nm. There is a preoscillatory or induction period, and the amplitude and the number of oscillations are associated with the initial concentration of reactants. The equations for the triiodide ion reaction rate changing with reaction time and the initial concentrations in the oscillation stage were obtained. Oscillation reaction can be accelerated by increasing temperature. The apparent activation energies in terms of the induction period and the oscillation period were 26.02 KJ/mol and 17.65 KJ/mol, respectively. The intermediates were detected by the online FTIR analysis. Based upon the experimental data in this work and in the literature, a plausible reaction mechanism was proposed for the oscillation reaction. PMID:22454614

  15. Chlorine Dioxide-Iodide-Methyl Acetoacetate Oscillation Reaction Investigated by UV-Vis and Online FTIR Spectrophotometric Method

    Laishun Shi


    Full Text Available In order to study the chemical oscillatory behavior and mechanism of a new chlorine dioxide-iodide ion-methyl acetoacetate reaction system, a series of experiments were done by using UV-Vis and online FTIR spectrophotometric method. The initial concentrations of methyl acetoacetate, chlorine dioxide, potassium iodide, and sulfuric acid and the pH value have great influence on the oscillation observed at wavelength of 289 nm. There is a preoscillatory or induction period, and the amplitude and the number of oscillations are associated with the initial concentration of reactants. The equations for the triiodide ion reaction rate changing with reaction time and the initial concentrations in the oscillation stage were obtained. Oscillation reaction can be accelerated by increasing temperature. The apparent activation energies in terms of the induction period and the oscillation period were 26.02 KJ/mol and 17.65 KJ/mol, respectively. The intermediates were detected by the online FTIR analysis. Based upon the experimental data in this work and in the literature, a plausible reaction mechanism was proposed for the oscillation reaction.

  16. Succinyl-CoA:acetoacetate transferase deficiency : identification of a new patient with a neonatal onset and review of the literature

    Niezen-Koning, K E; Wanders, R J; Ruiter, J P; Ijlst, L; Visser, G; Reitsma-Bierens, W C; Heijmans, Hugo; Reijngoud, D J; Smit, G P


    UNLABELLED: We describe the clinical symptoms and biochemical findings of a patient with succinyl-CoA:acetoacetate transferase deficiency who presented in the neonatal period and review the current literature on this subject. Our patient was initially suspected to have distal renal tubular acidosis,

  17. Succinyl-CoA : acetoacetate transferase deficiency: identification of a new patient with a neonatal onset and review of the literature

    NiezenKoning, KE; Ijlst, L; Visser, G; ReitsmaBierens, WCC; Heymans, HSA; Reijngoud, DJ; Smit, GPA; Ruiter, Jos P. N.


    We describe the clinical symptoms and biochemical findings of a patient with succinyl-CoA:acetoacetate transferase deficiency who presented in the neonatal period and review the current literature on this subject. Our patient was initially suspected to have distal renal tubular acidosis, and subsequ

  18. Metal-free arylation of ethyl acetoacetate with hypervalent diaryliodonium salts: an immediate access to diverse 3-aryl-4(1H)-quinolones.

    Monastyrskyi, Andrii; Namelikonda, Niranjan K; Manetsch, Roman


    A clean arylation protocol of ethyl acetoacetate was developed using hypervalent diaryliodonium salts under mild and metal-free conditions. The scope of the reaction, using symmetric and unsymmetric iodonium salts with varying sterics and electronics, was examined. Further, this method has been applied for the synthesis of antimalarial compound ELQ-300, which is currently in preclinical development.

  19. Effects of a novel allelochemical ethyl 2-methyl acetoacetate (EMA) on the ultrastructure and pigment composition of cyanobacterium Microcystis aeruginosa.

    Hong, Yu; Huang, Jing-Jing; Hu, Hong-Ying


    Allelochemical ethyl 2-methyl acetoacetate (EMA) can significantly inhibit the growth of bloom-forming Microcystis aeruginosa. In order to assess the implication of the damage of EMA on the algal photosynthetic apparatus, the effects of EMA on the algal ultrastructure and pigment composition were investigated. At initial exposure time to EMA (0-40 h), algal allophycocyanin, phycoerythrin and carotenoid degraded firstly; chlorophyll a increased, especially by 47% in the algae exposed to 2 mg L(-1) of EMA; phycocyanin was not significantly affected; lipid bodies increased remarkably. After 40 h of EMA exposure, chlorophyll a decreased gradually, especially by 45% in the algae exposed to 4 mg L(-1) of EMA; lipid bodies greatly reduced but cyanophycin granules accumulated; thylakoid structures were dissolved or disappeared with the presence of numerous vacuoles. These results showed that all ophycocyanin, phycoerythrin and carotenoid were more sensitive to EMA than other pigments, the cells of M. aeruginosa was stressed by EMA with the occurrence of cyanophycin granules and the photosynthesis pigments and ultrastructure of M. aeruginosa were quickly destroyed by EMA with exposure time increasing.

  20. Presolvated Electron Reactions with Methyl Acetoacetate: Electron Localization, Proton-Deuteron Exchange, and H-Atom Abstraction

    Alex Petrovici


    Full Text Available Radiation-produced electrons initiate various reaction processes that are important to radiation damage to biomolecules. In this work, the site of attachment of the prehydrated electrons with methyl acetoacetate (MAA, CH3-CO-CH2-COOCH3 at 77 K and subsequent reactions of the anion radical (CH3-CO•−-CH2-COOCH3 in the 77 to ca. 170 K temperature range have been investigated in homogeneous H2O and D2O aqueous glasses by electron spin resonance (ESR spectroscopy. At 77 K, the prehydrated electron attaches to MAA forming the anion radical in which the electron is delocalized over the two carbonyl groups. This species readily protonates to produce the protonated electron adduct radical CH3-C(•OH-CH2-COOCH3. The ESR spectrum of CH3-C(•OH-CH2-COOCH3 in H2O shows line components due to proton hyperfine couplings of the methyl and methylene groups. Whereas, the ESR spectrum of CH3-C(•OH-CH2-COOCH3 in D2O glass shows only the line components due to proton hyperfine couplings of CH3 group. This is expected since the methylene protons in MAA are readily exchangeable in D2O. On stepwise annealing to higher temperatures (ca. 150 to 170 K, CH3-C(•OH-CH2-COOCH3 undergoes bimolecular H-atom abstraction from MAA to form the more stable radical, CH3-CO-CH•-COOCH3. Theoretical calculations using density functional theory (DFT support the radical assignments.

  1. Hyperpolarized [1,3-13C2 ]ethyl acetoacetate is a novel diagnostic metabolic marker of liver cancer.

    Jensen, Pernille R; Serra, Sonia Colombo; Miragoli, Luigi; Karlsson, Magnus; Cabella, Claudia; Poggi, Luisa; Venturi, Luca; Tedoldi, Fabio; Lerche, Mathilde H


    An increased prevalence of liver diseases such as hepatitis C and nonalcoholic fatty liver results in an augmented incidence of the most common form of liver cancer, hepatocellular carcinoma (HCC). HCC is most often found in the cirrhotic liver and it can therefore be challenging to rely on anatomical information alone when diagnosing HCC. Valuable information on specific cellular metabolism can be obtained with high sensitivity thanks to an emerging magnetic resonance (MR) technique that uses 13C labeled hyperpolarized molecules. Our interest was to explore potential new high contrast metabolic markers of HCC using hyperpolarized 13C-MR. This work led to the identification of a class of substrates, low molecular weight ethyl-esters, which showed high specificity for carboxyl esterases and proved in many cases to possess good properties for signal enhancement. In particular, hyperpolarized [1,3-13C2 ]ethyl acetoacetate (EAA) was shown to provide a metabolic fingerprint of HCC. Using this substrate a liver cancer implanted in rats was diagnosed as a consequence of an ∼4 times higher metabolic substrate-to-product ratio than in the surrounding healthy tissue, (p=0.009). Unregulated cellular uptake as well as cosubstrate independent enzymatic conversion of EAA, made this substrate highly useful as a hyperpolarized 13C-MR marker. This could be appreciated by the signal-to-noise (SNR) obtained from EAA, which was comparable to the SNR reported in a literature liver cancer study with state-of-the-art hyperpolarized substrate, [1-13C]pyruvate. Also, the contrast-to-noise (CNR) in the EAA based metabolic ratio images was significantly improved compared with the CNR in equivalent images reported using [1-13C]pyruvate.

  2. Preparation and Catalytic Activity of PW12/PAn Material in Synthesis of 2 -Methyl-2-Ethyl Acetoacetate-1,3 -Dioxolane

    YANG Shui-jin; YU Xie-qing; LU Bao-lan; SUN Ju-tang


    Acetals and ketals are among the most important perfume materials and industrial materials of organic synthesis. Up to now, there are many methods to synthesize them.Conventionally H2SO4 is used as catalyst in factories, but it causes many problems, such as the erosion of equipment, difficulty for after-treatment, low quality of the products, etc. Heteropolyacids (HPA) and their salts have been extensively studied because of their interesting catalytic properties.Significant research effort has been directed towards the entrapment of heteropolyanions in suitable polymeric matrices with the main goal of preparing a new type of polymer-supported catalyst for various applications in heterogeneous and electrocatalysis. It showed that the main catalysis of HPA occurs through Brφnsted acid. The reports pointed that the rate of dehydration and etherification is directly in proportion to total acidity of heteropolyacids, HPA and its salts showed excellent catalytic activity to the esterification and have recently attracted much attention as catalysts for various industrial processes, because that their acidic and redox properties can be controlled at atomic/molecular levels. Mizuno and Misono, Pope, Kozhevnikov and Wang Enbo have reviewed the homogeneous catalysis and fine organic synthesis catalyzed by heteropoly compounds.Polyaniline (PAn) is another example of a polymer host which can accommodate anions originating from 12-tungstophosphoric acid (PW12). The polyemeraldine base was prepared by condensation-polymerization of aniline in aqueous HCl using (NH4)2S2O8 as the oxidant.PW12/PAn can be achieved by doping the protonation of the polyemeraldine base with H3PW12O40 in a two-step procedure. However, there is no report on the synthesis of 2-methyl-2-ethyl acetoacetate-1,3-dioxolane catalyzed by PW12/PAn.In this paper, 2-methyl-2-ethyl acetoacetate-1,3-dioxolane was synthesis from ethyl acetoacetate and ethylene glycol in the presence of PW12/PAn. The factors

  3. Responses of enzymatic antioxidants and non-enzymatic antioxidants in the cyanobacterium Microcystis aeruginosa to the allelochemical ethyl 2-methyl acetoacetate (EMA) isolated from reed (Phragmites communis).

    Hong, Yu; Hu, Hong-Ying; Xie, Xing; Li, Feng-Min


    Macrophytic allelochemicals are considered an environment-friendly and promising alternative to control algal bloom. However, studies examining the potential mechanisms of inhibitory allelochemicals on algae are few. The allelochemical ethyl 2-methyl acetoacetate (EMA), isolated from reed (Phragmites communis), was a strong allelopathic inhibitor on the growth of Microcystis aeruginosa. EMA-induced antioxidant responses were investigated in the cyanobacterium M. aeruginosa to understand the mechanism of EMA inhibition on algal growth. The activities of enzymatic antioxidants superoxide dismutase (SOD) and catalase (CAT), and the contents of non-enzymatic antioxidants reduced glutathione (GSH) and ascorbic acid (AsA) of M. aeruginosa cells were analyzed after treatments with different concentrations of EMA. Exposure of M. aeruginosa to EMA caused changes in enzyme activities and contents of non-enzymatic antioxidants in different manners. The decrease in SOD activity occurred first after 4 h of EMA exposure, and more markedly after 40 h. CAT activity did not change after 4 h of EMA exposure, but increased obviously after 40 h. The contents of AsA and GSH were increased greatly by EMA after 4 h. After 60 h, low EMA concentrations still increased the CAT activity and the contents of AsA and GSH, but high EMA concentrations started to impose a marked suppression on them. EMA increased dehydroascorbate (DHAsA) and oxidized glutathione (GSSG) contents during all exposure times. After 60 h, the regeneration rates of AsA and GSH (represented by the AsA/DHAsA ratio and GSH/GSSG ratio, respectively) were reduced by high EMA concentrations. These results suggest that the activation of CAT and the availability of AsA and GSH at early exposure are important to counteract the oxidative stress induced by EMA, and the inactivation of SOD may be crucial to the growth inhibition of M. aeruginosa by EMA.

  4. Research on Chemical Compositions of Acetoacetate in the Root of Gymnema Sylvestre%匙羹藤根部乙酸乙酯部分的化学成分研究

    姜建萍; 甄汉深; 曹音; 马雯芳; 林广莲; 王熙珍


    目的:对匙羹藤根乙酸乙酯部分的化学成分进行研究.方法:应用各种现代色谱手段对其化学成分进行分离纯化,利用理化性质结合光谱数据鉴定化合物结构.结果:分离鉴定了5个化合物,分别为正二十烷(Ⅰ)、豆甾醇(Ⅱ)、β-谷甾醇(Ⅲ)、正二十二烷酸(Ⅳ)、牛弥菜醇A(Ⅴ).结论:该方法能首次从该植物中分离得到化合物Ⅰ和Ⅳ.%Objective: To study the Chemical compositions of acetoacetate in the Root of Gymnema sylvestre. Methods: The compunds were isolated by many kinds of chromatography methods and identified on the basis of physico-chemical characters and spectroscopic analysis. Results: Five compounds were obtained from the ethyl acetate extract of Gymnema sylvestre (Retz.) Schult root, and their structures were identified as icosane (Ⅰ), Stigmasterol (Ⅱ), P-sitosterol (Ⅲ), docosanoic acid (Ⅳ) and conduritol A (Ⅴ). Conclusion: Compounds Ⅰ and Ⅳ are isolated from this plant for the first time.

  5. 乙酰乙酸/β羟丁酸检测用于1型和2型糖尿病分型的探讨%Study on the acetoacetate/β hydroxybutyrate determination in classification of type 1 and type 2 diabetes mellitus

    刘倩; 辛晓敏; 于永光; 金英玉; 王丽艳; 周游


    The clinical values of acetoacetate ( AcAA ) and β hydroxybutyrate ( βHBA ) determination in classification of type 1 and2 diabetes were explored. 102 normal control subjects,33 cases of type 1 diabetes, and 104cases of type 2 diabetes were enrolled. Serum AcAA, βHBA, fasting plasma glucose ( FPG), C-peptide, and insulin levels were measured. The results showed that serum AcAA, βHBA, total ketone tody (TKB) levels in the diabetic groups were significantly higher than those of the normal group( P<0. 01 ). AcAA, βHBA, TKB levels in type 1diabetes were higher as compared with those of type 2 diabetes( P<0.01 ). The AcAA, βHBA, and TKB levels were negatively related with C-peptide and insulin in diabetic patients( P<0. 01 ). All the type 1 diabetic patient were found to have TKB and lower C-peptide levels. TKB positive and lower C-peptide in type 2 diabetes were found in 47% and 26% respectively. Receiver operating characteristic (ROC) curve suggested that the area under the ROC curve of type 1 and type 2 diabetes was 0.926. The optimal operating point of the total ketone body was 0. 532 mmol/L with higher sensitivity and specificity. Enzymatic determination of acetoacetate and β hydroxybutyrate seems to have important clinical values for classification of type 1 and 2 diabetes.%探讨乙酰乙酸(AcAA)、β羟丁酸(βHBA)检测在1、2型糖尿病分型中的临床应用价值.收集健康体检者(正常对照组)102名,1型糖尿病患者(T1DM组)33例,2型糖尿病患者(T2DM组)104例,检测血AcAA、βHBA、空腹血糖(FPG)、C肽、胰岛素浓度.结果 显示,T1DM组和T2DM组AcAA、βHBA、AcAA+βHBA(总酮体,TKB)水平显著高于正常对照组,T1DM组显著高于T2DM组(均P<0.01);糖尿病患者(T1DM+T2DM)血AcAA、βHBA、TKB与C肽、胰岛素分别呈显著负相关(均P<0.01);T1DM组TKB阳性率100%、低C肽检出率100%,T2DM组TKB阳性率47%,低C肽检出率26%;通过建立受试者工作特征(ROC)曲线,T1DM组与T2DM

  6. Relationship between plasma concentration of β-hydroxybutyrate,acetoacetate, lactate and pyruvate and alcohol after oral or intravenous administration of alcohol to normal human subjects: a population pharmacodynamics (PD) analysis%口服及静注乙醇后血中乙醇与β-羟丁酸,乙酰乙酸,乳酸及丙酮酸浓度关系的探讨:一项群体药效动力学的研究

    万捷; 李建国; HUI C ko; Tom LIONETTI; David T GEORGE; Susan E SHOAF


    目的:应用群体药理学方法探讨血浆中乙醇浓度对β-羟丁酸,乙酰乙酸,乳酸,丙酮酸,β-羟丁酸/乙酰乙酸(H/A)比值及乳酸/丙酮酸(L/P)比值变化的效应.方法:给14名健康成人口服剂量相当于1.02 g·L-1总身体水的乙醇.在另一项实验中,给8名健康成人静脉注射剂量相当于0.83 g·L-1总身体水的乙醇.在服用乙醇后380 min采取静脉血测定乙醇,β-羟丁酸,乙酰乙酸,乳酸及丙酮酸的血浆浓度.在静注乙醇后340min采血测定上述5种物质的血浆浓度.结果:在口服乙醇实验中,G0为66.6±8.1 mg·dl-1,显著低于102mg·dl-1,(t检验,P<0.001).清除相斜率β为0.229±0.05 mg·dl-1·min-1.在静注实验中,G0为75.6±10.9 mg·dl-1,与83 mg·dl-1比较无显著性差异,β为0.245±0.05mg·dl-1·min-1.在两项实验中,我们应用群体间接生理反应模型来拟合乙醇浓度对β-羟丁酸,乙酰乙酸,乳酸,丙酮酸,β-羟丁酸/乙酰乙酸比值及乳酸/丙酮酸比值变化的效应,并得出各项参数.同时,我们发现,当乙醇的清除相结束时,H/A比值尚未达最大值,说明在乙醇的零级代谢相时肝脏仍在产生NADH.乳酸和乙醇的关系曲线显示乳酸的变化呈现一种逆时钟方向的滞后.结论:血L/P比值不适合用作实时肝脏氧化状态的指标.本研究提供的参数将有益于将来研究乙醇对肝脏氧化状态的影响.%To investigate the relationship between plasma concentration of β-hydroxybutyrate, acetoacetate, lactate, pyruvate, β-hydroxybutyrate/acetoacetate (H/A ratio) and lactate/pyruvate (L/P ratio) and alcohol by population PD analysis after oral or intravenous administration of alcohol. METHODS: An oral dose of alcohol tered to 14 normal human subjects and an Ⅳ infusion (30 water was administered to 8 normal subjects. Venous blood was sampled for determination of alcohol (BAC),β-hydroxybutyrate, acetoacetate, lactate and pyruvate for 380 min after oral administration and for

  7. Thermolytic reactions of esters. Part IV. The methyl esters of cyanoacetic, acetoacetic and malonic acid

    Tonkelaar, W.A.M. den; Louw, R.; Kooyman, E.C.


    The compounds on the title proved to be more stable than their ethyl analogues, 1,2-acid elimination being impossible. Acetic acid causes a pronounced rate enhancement. Kinetic as well as product data - including the effect of added water and/or alcohols - are interpreted on the basis of acetic acid

  8. 黄荆子乙酸乙酯提取物诱导人宫颈癌Hela细胞凋亡%Acetoacetate Extracte of Vitex Negundo Seed Induced Apoptosis in Hela Cell Line

    薛玉珍; 陈忠东


    [目的]探讨黄荊子乙酸乙酯提取物(EVn-50)体外诱导人宫颈癌Hela细胞的凋亡作用.[方法]体外培养Hela细胞,不同浓度EVn-50(1.0、10.0、100.0μg/ml)作用于Hela细胞.采用PI染色FCM检测EVn-50诱导Hela细胞的凋亡作用;采用间接免疫荧光标记FCM检测凋亡相关蛋白Bcl-2、Bax、Caspase-3的表达.[结果]EVn-50 1.0、10.0、100.0μg/ml处理Hela细胞48h后,细胞的凋亡率分别为1.06%、8.80%及20.90%,呈浓度依赖性(P<0.05).不同浓度EVn-50作用Hela细胞48h,细胞中Bax、Caspase-3蛋白表达逐渐升高,Bcl-2蛋白表达逐渐降低(P<0.05).[结论]EVn-50具有诱导Hela细胞凋亡作用,其作用可能与改变凋亡相关蛋白Bax、Caspase-3及Bcl-2的表达有关.

  9. Asymmetry Bioreduction of Ethyl Acetoacetate Catalyzed by Baker's Yeast%面包酵母催化乙酰乙酸乙酯的不对称还原反应

    朱文洲; 许建和; 俞俊棠



  10. 乙酸乙酯酯缩合法合成乙酰乙酸乙酯的热力学分析%Thermodynamics analysis of ethyl acetoacetate synthesis from ethyl acetate by ester condensation

    李丹; 丁同梅; 田恒水


    The formation enthalpy, Gibbs free energy and equilibrium constant for the reaction of ethyl acetate condensation to ethyl acetoacetatewere estimated by Benson group contribution method and Joback method. The results showed that, in 25 to 240℃, the formation enthalpy is positive, which indicated that the reaction is endothermic, and the heat absorption capacity decreases with the rise of temperature; the Gibbs free energy is positive, which indicated the reaction could not occur spontaneously; and the equilibrium constant is small but increases with the rise of temperature. So it required adding suitable catalysts to let the reaction take place smoothly, raising the reaction temperature to increase the conversion, or removing the products from the reaction system to promote the positive reaction.%用Benson基团贡献法和Joback法对乙酸乙酯酯缩合法生成乙酰乙酸乙酯反应的焓变,吉布斯自由能和反应平衡常数进行了计算与分析。结果显示,在25~240℃内,反应的焓变均为正,说明此反应为吸热反应,随着反应温度的升高吸热量减少;反应的吉布斯自由能一直为正,理论上是不能自发进行的;该反应的平衡常数很小,但是随着温度的升高,平衡常数相应有所增大。要使反应顺利进行需加入适当的催化剂,而为了提高反应的转化率,可以采取升高反应温度,或在反应过程中移走生成物的手段使反应不断向所需方向进行。

  11. Pseudoketogenesis in the perfused rat heart

    Fink, G.; Desrochers, S.; Des Rosiers, C.; Garneau, M.; David, F.; Daloze, T.; Landau, B.R.; Brunengraber, H.


    Ketogenesis is usually measured in vivo by dilution of tracers of (3R)-hydroxybutyrate or acetoacetate. We show that, in perfused working rat hearts, the specific activities of (3R)-hydroxybutyrate and acetoacetate are diluted by isotopic exchanges in the absence of net ketogenesis. We call this process pseudoketogenesis. When hearts are perfused with buffer containing 2.3 mM of (4-3H)- plus (3-14C)acetoacetate, the specific activities of (4-3H) and (3-14C)acetoacetate decrease while C-1 of acetoacetate becomes progressively labeled with 14C. This is explained by the reversibility of reactions catalyzed by mitochondrial 3-oxoacid-CoA transferase and acetoacetyl-CoA thiolase. After activation of labeled acetoacetate, the specific activity of acetoacetyl-CoA is diluted by unlabeled acetoacetyl-CoA derived from endogenous fatty acids or glucose. Acetoacetyl-CoA thiolase partially exchanges 14C between C-1 and C-3 of acetoacetyl-CoA. Finally, 3-oxoacid-CoA transferase liberates weakly labeled acetoacetate which dilutes the specific activity of extracellular acetoacetate. An isotopic exchange in the reverse direction is observed when hearts are perfused with unlabeled acetoacetate plus (1-14C)-, (13-14C)-, or (15-14C)palmitate; here also, acetoacetate becomes labeled on C-1 and C-3. Computations of specific activities of (3R)-hydroxybutyrate, acetoacetate, and acetyl-CoA yield minimal rates of pseudoketogenesis ranging from 19 to 32% of the net uptake of (3R)-hydroxybutyrate plus acetoacetate by the heart.

  12. Determination of ketone bodies in blood by headspace gas chromatography-mass spectrometry

    Holm, Karen Marie Dollerup; Linnet, Kristian; Rasmussen, Brian Schou


    A gas chromatography-mass spectrometry (GC-MS) method for determination of ketone bodies (ß-hydroxybutyrate, acetone, and acetoacetate) in blood is presented. The method is based on enzymatic oxidation of D-ß-hydroxybutyrate to acetoacetate, followed by decarboxylation to acetone, which...... was quantified by the use of headspace GC-MS using acetone-(13)C(3) as an internal standard. The developed method was found to have intra- and total interday relative standard deviations acetone+acetoacetate levels (~25 to 8300 µM) and D-ß-hydroxybutyrate levels (~30 to 16500 µM). Recovery values varied...

  13. Synthesis of 1, 4-Dihydropyridine Derivatives using FeCl3 as Catalyst under Solvent-free Condition



    Full Text Available A mixture of ethyl acetoacetate, benzaldehyde and ammonium acetate and in the presence of FeCl3 under solvent-free condition were converted to 1, 4-dihydropyridines with good yields.

  14. A New Strategy for the Synthesis of 3-Acyl-coumarin Using Mesoporous Molecular Sieve MCM-41 as a Novel and Efficient Catalyst



    3-Acyl-coumarins were obtained in high yields from ortho-hydroxybenzaldehydes and ethyl acetoacetate or ethyl benzoylacetate in acetonitrile in the presence of a catalytic amount of mesoporous molecular sieve MCM-41. 3-Acyl-coumarins were obtained in high yields from ortho-hydroxybenzaldehydes and ethyl acetoacetate or ethyl benzoylacetate in acetonitrile in the presence of a catalytic amount of mesoporous molecular sieve MCM-41.

  15. Ketogenesis in muscle: artifact or reality

    Fink, G.; Desrochers, S.; Garneau, M.; David, F.; Landau, B.R.; Brunengraber, H.


    It has been proposed that muscle is the site of net ketogenesis. This hypothesis was based on a discrepancy between the balance of unlabeled and labeled ketone bodies across muscle beds in humans infused with (/sup 14/C)acetoacetate. It has been pointed out that the dilution of the specific activity of acetoacetate could be explained by the reversal of 3-oxoacid-CoA transferase. Catabolism of endogenous or exogenous fatty acids dilutes the specific activities of acetoacetyl-CoA and, presumably, of acetoacetate. The latter mechanism was tested in perfused working hearts and hemicorpi of rats. Organs were perfused with mM concentrations of (4-/sup 3/H)acetoacetate. The specific activities of perfusate acetoacetate and 3-hydroxybutyrate equilibrated rapidly, and then decreased 15-20% over 90 min. There was net uptake of ketone bodies. The authors conclude that (i) the data purporting to show ketogenesis in muscle can be explained by isotopic exchange between acetoacetate and acetoacetyl-CoA, and (ii) there is no net ketogenesis in muscle.

  16. Ferric Perchlorate Catalyzed One-pot Synthesis of 1,2,3,4-Tetrahydro-2-pyrimidinones and -thiones:an Expedient Protocol for the Biginelli Reaction

    HERAVI,Majid Momahed; BEHBAHANI,Farahnaz Kargar OSKOOIE; Hossien Abdi


    An efficient synthesis of 1,2,3,4-tetrahydro-2-pyrimidinones and -thiones using ferric perchlorate as the catalyst from an aldehyde, ethyl acetoacetate, and urea or thiourea in acetonitrile was described. Compared to the classical Biginelli reaction conditions, this new method consistently has the advantage of full catalysis, good yields and short reaction time.

  17. L-proline-catalysed synthesis of functionalized unsymmetrical dihydro-1H-indeno[1,2-b]pyridines

    Farahnaz Kargar Behbahani; Hourieh Sadat Alaei


    Aromatic aldehydes have been employed in a one-pot four-component reaction with ethyl acetoacetate, 1,3-indandione and ammonium acetate in the presence of L-proline in water and under reflux condition to afford the corresponding dihydro-1H-indeno[1,2-b]pyridines in very good yields.

  18. 21 CFR 172.515 - Synthetic flavoring substances and adjuvants.


    .... Isoquinoline. Isovaleric acid. cis-Jasmone; 3-methyl-2-(2-pentenyl)-2-cyclopenten-1-one. Lauric aldehyde...; methyl ethyl ketone. Butter acids. Butter esters. Butyl acetate. Butyl acetoacetate. Butyl alcohol; 1-butanol. Butyl anthranilate. Butyl butyrate. Butyl butyryllactate; lactic acid, butyl ester, butyrate....

  19. Pechmann Reaction Promoted by Boron Trifluoride Dihydrate

    J. Mezger


    Full Text Available The Pechmann reaction of substituted phenols 1a-e with methyl acetoacetate (2 can be activated by boron trifluoride dihydrate (3 to give the corresponding 4-methyl- coumarin derivatives 4a-e in excellent yield (98-99 %.

  20. Silica (NPs) supported Fe (III) as a reusable heterogeneous catalyst for the one-pot synthesis of 1, 4-dihydropyridines under mild conditions

    Javad Safaei-Ghomi; Abolfazl Ziarati; Safura Zahedi


    A cheap and recyclable silica (NPs) supported Fe (III) was prepared as heterogeneous catalyst for the synthesis of various substituted 1,4-dihydropyridines via condensation of aldehydes with ethyl acetoacetate and ammonium acetate in ethanol. The products were separated from the catalyst simply by filtration and the catalyst could be recycled and reused for several times without noticeable decrease in the catalytic activity.

  1. ZnO Nanoparticles: An Efficient Reagent, Simple and One-Pot Procedure for Synthesis of Highly Functionalized Dihydropyridine Derivatives

    Marziyeh Khazaei


    Full Text Available A new and efficient one-pot synthesis of dihydropyridones derivatives by four-component reaction between cyanoacetamide, aryl aldehydes, and ethyl acetoacetate with ammonium acetate using nano ZnO is described. The reaction was performed in ethanol under reflux conditions and afforded good yields of products.

  2. Effects of a single glucocorticoid injection on propylene glycol-treated cows with clinical ketosis

    Drift, van der S.G.A.; Houweling, M.; Bouman, Marina; Koets, A.P.; Tielens, A.G.M.; Nielen, M.; Jorritsma, R.


    This study investigated the metabolic effects of glucocorticoids when administered to propylene glycol-treated cows with clinical ketosis. Clinical ketosis was defined by depressed feed intake and milk production, and a maximal score for acetoacetate in urine. All cows received 250 mL oral propylene

  3. Synthesis of 3-cyano-4-aryl-5-ethoxycarbonyl-6-methyl-3,4-dihydropyridine-2-thiones

    Krauze, A.A.; Liepin' sh, E.E.; Pelcher, Yu.E.; Kalme, Z.A.; Dipan, I.V.; Dubur, G.Ya.


    The condensation of ethyl arylidenacetoacetate with cyanothioacetamide and of arylidenecyanothioacetamides with ethyl acetoacetate or of arylidenecyanothioacetamides with ethyl ..beta..-aminocrotonate gave 3-cyano-4-aryl-5-ethoxycarbonyl-6-methyl-3,4-dihydropyridine-2-thiones. PMR spectroscopy showed that the 3-cyano-4-aryl-3,4-dihydro-pyridine-2-thiones are formed as a mixture of cis and trans isomers.

  4. Poly(4-vinylpyridine efficiently catalyzed one-pot four-component synthesis of pyrano[2,3-c]pyrazoles

    Jalal Albadi


    Full Text Available An efficient one-pot synthesis of pyrano[2,3-c]pyrazoles via four-component reaction of phenyl hydrazine, ethyl acetoacetate, malononitrile and aromatic aldehydes, catalyzed by poly(4-vinylpyridine is reported. This method provides many advantages such as, atom-economy, easy work up, clean procedure, short reaction times and high yields of products.

  5. A Novel and Efficient Five-Component Synthesis of Pyrazole Based Pyrido[2,3-d]pyrimidine-diones in Water: A Triply Green Synthesis

    Majid M. Heravi


    Full Text Available A novel one pot synthesis of pyrazolo[4′,3′:5,6]pyrido[2,3-d]pyrimidine-diones, via a five-component reaction, involving, hydrazine hydrate, ethyl acetoacetate, and 1,3-dimethyl barbituric acid, an appropriate aryl aldehydes and ammonium acetate catalyzed via both of heterogeneous and homogeneous catalysis in water, is reported.

  6. Methyl 6-Methyl-1-(4-methylphenyl-2-oxo-4-phenyl-1,2,3,4-tetrahydropyrimidine-5-carboxylate

    Haiping Wang


    Full Text Available Methyl 6-methyl-1-(4-methylphenyl-2-oxo-4-phenyl-1,2,3,4-tetrahydro-pyrimidine-5-carboxylate has been synthesized via the modified Biginelli reaction from benzaldehyde, p-tolylurea, and methyl acetoacetate, promoted with microwave irradiation and catalyzed by TsOH under solvent-free conditions in high yield.

  7. Ketogenesis evaluation in perfused liver of diabetic rats submitted to short-term insulin-induced hypoglycemia.

    Barrena, Helenton Cristhian; Gazola, Vilma Aparecida Ferreira Godoi; Furlan, Maria Montserrat Diaz Pedrosa; Garcia, Rosângela Fernandes; de Souza, Helenir Medri; Bazotte, Roberto Barbosa


    Ketogenesis, inferred by the production of acetoacetate plus ss-hydroxybutyrate, in isolated perfused livers from 24-h fasted diabetic rats submitted to short-term insulin-induced hypoglycemia (IIH) was investigated. For this purpose, alloxan-diabetic rats that received intraperitoneal regular insulin (IIH group) or saline (COG group) injection were compared. An additional group of diabetic rats which received oral glucose (gavage) (100 mg kg(-1)) 15 min after insulin administration (IIH + glucose group) was included. The studies were performed 30 min after insulin (1.0 U kg(-1)) or saline injection. The ketogenesis before octanoate infusion was diminished (p ketogenesis, livers from diabetic rats submitted to short-term IIH which received insulin or insulin plus glucose showed maintained capacity to produce acetoacetate and ss-hydroxybutyrate from octanoate.

  8. Nano Fe2O3, clinoptilolite and H3PW12O40 as efficient catalysts for solvent-free synthesis of 5(4H)-isoxazolone under microwave irradiation conditions

    Fozooni,Samieh; Hosseinzadeh,Nasrin Gholam; Hamidian, Hooshang; Akhgar,Mohammad Reza


    A quick and solvent-free approach involving the exposure of neat reactants to microwave irradiation in conjunction with the use of clinoptilolite, H3PW12O40 and Fe2O3 nanoparticle catalysts is described. In this work, condensation of hydroxylamine hydrochloride, sodium acetate, acetoacetic or benzoyl acetic ethyl ester and appropriate aldehydes by employing catalysts gave 5(4H)-isoxazolone only in one step. Catalyst amount, temperature effects and catalysts reusability were monitored. Among t...

  9. Synthesis of 2-azetidinones substituted coumarin derivative

    Jha Mukesh S.


    Full Text Available α-Naphthol is converted into 4-methylbenzo[h]chromen-2-one by reacting with ethyl acetoacetate in the presence of bismuth trichloride which is then oxidized to 2-oxo-2H-benzo[h]chromene-4-carbaldehyde and then condensed with aromatic primary amines to give Schiff bases (3a-3d. These Schiff bases are then reacted with acid chlorides in the presence of base in toluene to give 1, 3, 4-substituted 2-azetidinones.

  10. PPA-SiO2 catalyzed efficient synthesis of polyhydroquinoline derivatives through Hantzsch multicomponent condensation under solvent-free conditions

    Amir Khojastehnezhad; Farid Moeinpour; Abolghasem Davoodnia


    Silica gel-supported polyphosphoric acid (PPA-SiO2) was found to be an efficient catalyst for the one-pot four-component Hantzsch condensation reaction of aryl aldehydes, dimedone, ethyl acetoacetate and ammonium acetate to afford the corresponding polyhydroquinoline derivatives in high yields. The main advantages of the present approach are short reaction times, clean reaction profiles, simple experimental and workup procedures.

  11. Ketone body β-hydroxybutyrate blocks the NLRP3 inflammasome-mediated inflammatory disease

    Youm, Yun-Hee; Nguyen, Kim Y.; Grant, Ryan W.; Goldberg, Emily L.; Bodogai, Monica; Kim, Dongin; D'Agostino, Dominic; Planavsky, Noah; Lupfer, Christopher; Kanneganti, Thirumala D.; Kang, Seokwon; Horvath, Tamas L.; Fahmy, Tarek M.; Crawford, Peter A.; Biragyn, Arya


    Ketone bodies , β-hydroxybutyrate (BHB) and acetoacetate support mammalian survival during states of energy deficit by serving as alternative source of ATP 1 . BHB levels are elevated during starvation, high-intensity exercise or by the low carbohydrate ketogenic diet 2 . Prolonged caloric restriction or fasting reduces inflammation as immune system adapts to low glucose supply and energy metabolism switches towards mitochondrial fatty acid oxidation, ketogenesis and ketolysis 2-6 . However, ...

  12. MgO Nanoparticles as a Recyclable Heterogeneous Catalyst for the Synthesis of Polyhydroquinoline Derivatives under Solvent Free Conditions%MgO Nanoparticles as a Recyclable Heterogeneous Catalyst for the Synthesis of Polyhydroquinoline Derivatives under Solvent Free Conditions

    Ranjbar-Karimi, Reza; Hashemi-Uderji, Somayeh; Bazmandegan-Shamili, Alireza


    Magnesium oxide nanopartticels in average size between 35--120 nm were prepared by sonochemistry method. Synthesis of polyhydroquinoline derivatives using MgO nanoparticles from the reaction of dimedone, benzaldehyde ethyl acetoacetate and ammonium acetate under solvent-free conditions is reported. Easy handling, reusability, thermal stability and non-toxicity of the catalyst make the present protocol as an eco-friendly and economically acceptable method for synthesis of these heterocycles.

  13. One-pot, four-component synthesis of pyrano[2,3-c]pyrazoles catalyzed by sodium benzoate in aqueous medium

    Hamzeh Kiyani


    Full Text Available An efficient, green, and facile four-component reaction for the preparation of pyrano[2,3-c]pyrazole derivatives through the condensation reaction of aryl aldehydes, ethyl acetoacetate, malononitrile, and hydrazine hydrate or phenyl hydrazine in the presence of commercially available organocatalyst sodium benzoate under aqueous condition is reported. The products are produced with high yields and in shorter reaction times. It also is mild, safe, green and environmental friendly.

  14. Cerium(Ⅳ) ammonium nitrate catalysed facile and efficient synthesis of polyhydroquinoline derivatives through Hantzsch multicomponent condensation

    Cherkupally Sanjeeva Reddy; Mekala Raghu


    A general and convenient practical approach for the synthesis of polyhydroquinoline derivatives has been achieved via one-pot four-component Hantzsch condensation of aromatic aldehydes,dimedone,ethyl acetoacetate and ammonium acetate in the presence of a catalytic amount of cerium(Ⅳ) ammonium nitrate (CAN),in ethanol solvent at ambient temperature.Simple work-up,mild reaction conditions,inexpensive and non-toxic catalyst,and excellent product yields are the advantageous features of this method.

  15. Mesostructured SBA-15-Pr-SO3H: An efficient solid acid catalyst for one-pot and solvent-free synthesis of 3,4-dihydro-2-pyridone derivatives

    Ghodsi Mohammadi Ziarani; Somayeh Mousavi; Negar Lashgari; Alireza Badiei


    3,4-Dihydro-2-pyridone derivatives have been prepared efficiently via a one-pot four-component reaction of benzaldehyde derivatives, Meldrum’s acid, methyl acetoacetate and ammonium acetate in the presence of sulphonic acid-functionalized ordered nanoporous SBA-15 as a nano heterogeneous catalyst under solvent-free conditions. This process is a simple, environmentally friendly, rapid and high yielding reaction.

  16. Solid-supported sulfonic acid-containing catalysts efficiently promoted one-pot multi-component synthesis of -acetamido carbonyl compounds

    Mohammad Ali Zolfigol; Ardeshir Khazaei; Abdolkarim Zare; Mohammad Mokhlesi; Tahereh Hekmat-Zadeh; Alireza Hasaninejad; Fatemeh Derakhshan-Panah; Ahmad Reza Moosavi-Zare; Hassan Keypour; Ahmad Ali Dehghani-Firouzabadid; Maria Merajoddin


    Silica-functionalized sulfonic acid (SFSA) and sulfuric acid-modified polyethylene glycol-6000 (PEG-OSO3H) efficiently catalysed one-pot multi-component condensation of enolizable ketones or alkyl acetoacetates with arylaldehydes, acetonitrile and acetyl chloride to afford the corresponding -acetamido ketone or ester derivatives in high to excellent yields and in relatively short reaction times. Moreover, in this work, some novel -acetamido carbonyl compounds (i.e., one complex structure) are synthesized.

  17. Expression of Clostridium acetobutylicum ATCC 824 Genes in Escherichia coli for Acetone Production and Acetate Detoxification

    Bermejo, Lourdes L.; Welker, Neil E.; Papoutsakis, Eleftherios T.


    A synthetic acetone operon (ace4) composed of four Clostridium acetobutylicum ATCC 824 genes (adc, ctfAB, and thl, coding for the acetoacetate decarboxylase, coenzyme A transferase, and thiolase, respectively) under the control of the thl promoter was constructed and was introduced into Escherichia coli on vector pACT. Acetone production demonstrated that ace4 is expressed in E. coli and resulted in the reduction of acetic acid levels in the fermentation broth. Since different E. coli strains...

  18. Ketone Body Metabolic Enzyme OXCT1 Regulates Prostate Cancer Chemoresistance


    Fig. 8). Taken together, the increased oxygen consumption but decreased ATP and NADH production indicated that OXCT1 knock down induced metabolic... method . shC or shOXCT1 cells without Li-Acetoacetate treatment were set as 100, respectively. Progress and Accomplishments We established OXCT1...Nelson PS, Beer TM. Prostate cancer-associated gene expression alterations determined from needle biopsies. Clin Cancer Res 15 (9): 3135-42, 2009. 2

  19. Influence of Electronic Factors on “Solvent-Free and Catalyst-Free Biginelli Reaction

    Tanay Pramanik


    Full Text Available A series of Biginelli condensation were carried out employing urea, ethyl acetoacetate with a series of different derivatives of benzaldehyde. A green synthetic procedure was followed for performing the Biginelli reactions. The reaction conditions were found to be different for electron rich and electron deficient benzaldehydes. So the electronic effect has shown a significant impact on the reaction condition of the Biginelli condensation.

  20. Synthesis of Polyfunctionalized 4H-Pyrans

    Manisha Bihani


    Full Text Available Amberlyst A21 catalyzed one-pot three-component coupling of aldehyde and malononitrile with active methylene compounds such as acetylacetone and ethyl acetoacetate for the synthesis of pharmaceutically important polyfunctionalized 4H-pyrans has been reported. Simple experimental procedure, no chromatographic purification, no hazardous organic solvents, easy recovery and reusability of the catalyst, and room temperature reaction conditions are some of the highlights of this protocol for the synthesis of pharmaceutically relevant focused libraries.

  1. Synthesis of Some New Benzimidazole Derivatives of Pharmaceutical Interest

    Fawzia Zakaria El-Ablack


    Reaction of 2-(aminomethyl)benzimidazole dihydrochloride (1) with ethyl acetoacetate was studied to give diazepinone-benzimidazole derivative (2), while, treatment of 1 with phenylhydrazono ethylacetoacetate afforded phenylhydrazino diazepinone derivative (3). On the other hand, reaction of 1 with acetyl acetone resulted in the formation of diazepine derivative (4). The reaction of 1 with ethyl cyanoacetate was studied to give 3-aminodiazepinone derivative (5). Also the reaction of 1 with ace...

  2. Condensation Reaction of 5, 6-Dihydro-6-methyl-6-piperonyl-2H- pyran-2,4-dione, Ethyl Orthoformate and Substituted Anilines


    Piperonyl methyl ketone was obtained by oxidizing isosafrole with hydrogen peroxide and formic acid. Dianion of ethyl acetoacetate reacted with piperonyl methyl ketone and 5, 6-dihydro-6-methyl-6-piperonyl-2H-pyran-2, 4-dione was prepared, which reacted with substituted anilines in the presence of ethyl orthoformate to obtain 3-anilinomethylene-5, 6- dihydro- 6-methyl-6-piperonyl-2H-pyran-2, 4-diones. Their structures were confirmed by 1HNMR and elemental analysis.

  3. Synthesis of Some New 1,4-Dihydropyridine Derivatives through a Facile One-pot Hantzsch Condensation Catalyzed by Triethylamine

    Ghalem, Wassima; Boulcina, Raouf; Debache, Abdelmadjid


    A facile and efficient synthesis of new 1,4-dihydropyridine derivatives was reported via Hantzsch three-com- ponent condensation reaction of aldehydes or formylphenylboronic acids, ethyl acetoacetate, and ammonium acetate in the presence of a catalytic amount of triethylamine under solvent-free conditions. The method described here of- fers several advantages including high yields, short reaction times, and simple work-up procedure.

  4. Efficient Synthesis of β-Acetamido Ketones and Esters Using Aluminum Chloride as an Inexpensive and Green Catalyst%Efficient Synthesis of β-Acetamido Ketones and Esters Using Aluminum Chloride as an Inexpensive and Green Catalyst

    Mohammad Ali, Zolfigol; Ardeshir, Khazaei; Mohammad, Mokhlesi; Abdolkarim, Zare; Maliheh, Safaiee; Fatemeh, Derakhshan-Panah; Hassan, Keypour; Ahmad, Ali Dehghani-Firouzabadi; Maria, Merajoddin


    Aluminum chloride (AlCl3) efficiently catalyzes one-pot multicomponent condensation of enolizable ketones or alkyl acetoacetates with aldehydes, acetonitrile and acetyl chloride to afford β-acetamido ketone or ester derivatives in high to excellent yields and in relatively short reaction times. Moreover, by this synthetic method, some novel β-acetamido ketones and esters (i.e. one complex structure) are prepared.

  5. Snail shell as a new natural and reusable catalyst for synthesis of 4H-Pyrans derivatives

    Zakaria Benzekri


    Full Text Available A simple, efficient and general method for the synthesis of 4H-pyrans is established through a multi component cyclocondensation of aromatic aldehydes, malononitrile and ethyl acetoacetate or acetyl acetone using snail shell as a natural catalyst. In this method the snail shell was used as green and reusable natural catalyst. Excellent yields, short reaction times and availability of the catalyst are the advantages of this method.

  6. Purification and characterization of a novel carbonyl reductase with high stereo-selectivity

    YANG Ming; XU Yan; MU Xiaoqing; XIAO Rong


    A novel NADPH-dependent carbonyl reductase was separated from Candida parapsilosis CCTCC 203011.The enzyme gave a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE),which was purified through ammonium sulfate,Diethylamino Ethanol (DEAE) sepharose Fast flow (FF),phenyl-sepharose FF and blue sepharose FF chromatography from cell-free extract.The molecular mass of the enzyme was about 30 kDa.The optimum pH and temperature for reduction were 4.5℃ and 35℃,respectively.The Cu2+ had strong restrictive effect on enzyme activity.In addition,the carbonyl reductase was an enzyme with high substrate specificity and stereo-selectivity,and showed high asymmetric reduction activity towards α-hydroxyacetophenone and ethyl 4-chloro acetoacetate.For the asymmetric reduction of α-hydroxyacetophenone and ethyl 4-chloro acetoacetate,(S)-1-phenyl-1,2-ethanediol and (R)-ethyl 4-chloro-3-hydroxybutanoate were produced by the purified enzyme,with the 100% and 94.3% e.e.value,respectively.Therefore,the enzyme could be one of the effective biocatalysts for asymmetric synthesis of chiral alcohols.The amino acid sequences of one peptide from the purified enzyme were analyzed by LC-MASS-MASS,and the carbonyl reductase showed some identity to the hypothetical protein CaO 19.10414 reported.

  7. Metabolic engineering of Clostridium tyrobutyricum for n-butanol production: effects of CoA transferase.

    Yu, Le; Zhao, Jingbo; Xu, Mengmeng; Dong, Jie; Varghese, Saju; Yu, Mingrui; Tang, I-Ching; Yang, Shang-Tian


    The overexpression of CoA transferase (ctfAB), which catalyzes the reaction: acetate/butyrate + acetoacetyl-CoA → acetyl/butyryl-CoA + acetoacetate, was studied for its effects on acid reassimilation and butanol biosynthesis in Clostridium tyrobutyricum (Δack, adhE2). The plasmid pMTL007 was used to co-express adhE2 and ctfAB from Clostridium acetobutylicum ATCC 824. In addition, the sol operon containing ctfAB, adc (acetoacetate decarboxylase), and ald (aldehyde dehydrogenase) was also cloned from Clostridium beijerinckii NCIMB 8052 and expressed in C. tyrobutyricum (Δack, adhE2). Mutants expressing these genes were evaluated for their ability to produce butanol from glucose in batch fermentations at pH 5.0 and 6.0. Compared to C. tyrobutyricum (Δack, adhE2) without expressing ctfAB, all mutants with ctfAB overexpression produced more butanol, with butanol yield increased to 0.22 - 0.26 g/g (vs. 0.10 - 0.13 g/g) and productivity to 0.35 g/l h (vs. 0.13 g/l h) because of the reduced acetate and butyrate production. The expression of ctfAB also resulted in acetone production from acetoacetate through a non-enzymatic decarboxylation.

  8. Fatty acid oxidation and ketogenesis in astrocytes

    Auestad, N.


    Astrocytes were derived from cortex of two-day-old rat brain and grown in primary culture to confluence. The metabolism of the fatty acids, octanoate and palmitate, to CO{sub 2} in oxidative respiration and to the formation of ketone bodies was examined by radiolabeled tracer methodology. The net production of acetoacetate was also determined by measurement of its mass. The enzymes in the ketogenic pathway were examined by measuring enzymic activity and/or by immunoblot analyses. Labeled CO{sub 2} and labeled ketone bodies were produced from the oxidation of fatty acids labeled at carboxy- and {omega}-terminal carbons, indicating that fatty acids were oxidized by {beta}-oxidation. The results from the radiolabeled tracer studies also indicated that a substantial proportion of the {omega}-terminal 4-carbon unit of the fatty acids bypassed the {beta}-ketothiolase step of the {beta}-oxidation pathway. The ({sup 14}C)acetoacetate formed from the (1-{sup 14}C)labeled fatty acids, obligated to pass through the acetyl-CoA pool, contained 50% of the label at carbon 3 and 50% at carbon 1. In contrast, the ({sup 14}C)acetoacetate formed from the ({omega}-1)labeled fatty acids contained 90% of the label at carbon 3 and 10% at carbon 1.

  9. Study of ketone body kinetics in children by a combined perfusion of /sup 13/C and /sup 2/H3 tracers

    Bougneres, P.F.; Ferre, P.


    Ketone body kinetics were quantified in six children (3-5 yr old), who were fasted for 13-22 h, by a combined perfusion of (3-/sup 13/C)acetoacetate ((/sup 13/C)AcAc) and D-(-)-beta-(4,4,4-/sup 2/H3)hydroxybutyrate (beta-(/sup 2/H3)OHB) and gas chromatography-mass spectrometry analysis. Results were analyzed according to the single-pool (combined enrichments) or the two-accessible pools models. After 20-22 h of fasting, ketone body turnover rate was 30-50, a rate achieved after several days of fasting in adults. At low ketosis, acetoacetate was the ketone body preferentially synthesized de novo and utilized irreversibly. When ketosis increased, acetoacetate irreversible disposal was not enhanced, since it was largely converted into beta-OHB, whereas beta-OHB irreversible disposal was very much increased. The single-pool and two-pool models gave similar ketone body turnover rates when (/sup 13/C)AcAc was the tracer, whereas the use of beta-(/sup 2/H3)OHB gave some more divergent results, especially at low ketosis. These studies demonstrate that ketogenesis is very active in short-term fasted children and that the use of a combined infusion of (/sup 13/C)AcAc and beta-(/sup 2/H3)OHB is a convenient way to give insight into individual ketone body kinetics.

  10. Plasma carnitine concentration and lipid metabolism in infants receiving parenteral nutrition.

    Christensen, M L; Helms, R A; Mauer, E C; Storm, M C


    The relationships among plasma total carnitine concentration, postnatal age, and fatty acid metabolism were evaluated in 57 infants receiving parenteral nutrition. Concentrations of plasma carnitine, triglycerides, free fatty acids, acetoacetate, and beta-hydroxybutyrate were determined before and at 2 and 4 hours from the beginning of a standardized 2-hour lipid infusion. Plasma carnitine concentrations declined with increasing postnatal age. There were no significant differences in gestational age or triglyceride concentrations between infants less than or equal to 4 weeks of age and those greater than 4 weeks of age, whereas free fatty acid concentrations were lower and acetoacetate and beta-hydroxybutyrate concentrations were higher in the younger infants. Infants less than or equal to 4 weeks of age were further grouped according to plasma carnitine concentration greater than 13 nmol/ml (group 1) and less than or equal to 13 nmol/ml (group 2) and were then compared with infants greater than 4 weeks of age (group 3). There were no significant differences in triglyceride concentrations among the three groups; free fatty acids, acetoacetate, and beta-hydroxybutyrate concentrations for group 2 patients were similar to those of group 1 patients or fell between values for group 1 and group 3 patients. These results demonstrate decreasing plasma carnitine concentrations and possibly for more than 4 weeks.

  11. Control of ketogenesis in the perfused rat liver by the sympathetic innervation.

    Beuers, U; Beckh, K; Jungermann, K


    The regulation of ketogenesis by the hepatic nerves was investigated in the rat liver perfused in situ. Electrical stimulation of the hepatic nerves around the portal vein and the hepatic artery caused a reduction of basal ketogenesis owing to a decrease in acetoacetate release to 30% with essentially no change in 3-hydroxybutyrate release. At the same time, as observed before [Hartmann et al. (1982) Eur. J. Biochem. 123, 521-526], nerve stimulation increased glucose output, shifted lactate uptake to output and decreased perfusion flow. Ketogenesis from oleate, which enters the mitochondria via the carnitine system, was also lowered after nerve stimulation owing to a decrease of acetoacetate release to 30% with no alteration in 3-hydroxybutyrate release. Ketogenesis from octanoate, which enters the mitochondria independently of the carnitine system, was decreased after nerve stimulation as a result of a drastic decrease of acetoacetate output to 15% and a less pronounced decrease of 3-hydroxybutyrate release to 65%. Noradrenaline mimicked the metabolic nerve effects on ketogenesis only at the highly unphysiological concentration of 0.1 microM under basal conditions and in the presence of oleate as well as partly in the presence of octanoate. It was essentially not effective at a concentration of 0.01 microM, which might be reached in the sinusoids owing to overflow from the hepatic vasculature. Sodium nitroprusside prevented the hemodynamic changes after nerve stimulation; it did not affect the nerve-dependent reduction of ketogenesis under basal conditions and in the presence of oleate, yet it diminished the nerve effect on octanoate-dependent ketogenesis. Phentolamine clearly reduced the metabolic and hemodynamic nerve effects, while propranolol was without effect. The present data suggest that hepatic ketogenesis was inhibited by stimulation of alpha-sympathetic liver nerves directly rather than indirectly via hemodynamic changes or noradrenaline overflow from

  12. A green synthesis of α,β-unsaturated carbonyl compounds from glyceraldehyde acetonide

    Cláudia O. Veloso


    Full Text Available The catalytic behavior of Cs-exchanged and Cs-impregnated zeolites (X and Y was studied using the Knoevenagel condensation between glyceraldehyde acetonide and ethyl acetoacetate in order to produce the corresponding α,β-unsaturated carbonyl compound that is an important intermediate for fine chemicals. The influence of reaction temperature, type of zeolite, and basicity of the sites on the catalytic behavior of the samples was evaluated. All zeolites were active for the studied reaction. The formation of the main condensation product was favored at lower reaction temperatures. Products of further condensations were also observed especially for samples that were only dried before catalytic test.

  13. Nano-TiCl4/SiO2: An efficient heterogeneous solid acid catalyst for the one pot cascade five-component synthesis of densely functionalized tetrahydropyridines

    Abdolhamid Bamoniri; Bi Bi Fatemeh Mirjalili; Reza Tarazian


    Nano-TiCl4/SiO2 was found to be an inexpensive and efficient heterogeneous solid acid catalyst for the synthesis of one-pot cascade synthesis of highly functionalized asymmetric tetrahydropyridines from the five-component condensation reaction of the para-substituted anilines and aromatic aldehydes with ethyl acetoacetate under thermal conditions. Novel methodology, environmentally benign conditions, clean protocol, easy work-up and high yields are the important advantages of this protocol. The products were characterized using physical and spectroscopic data such as FT-IR, 1H-NMR and in some cases 13C-NMR and CHN analysis.

  14. Intense Photoluminescence and Movement of OpticalAbsorption Edge in Alumina Aerogels


    Alumina aerogels with high porosity of nanometer size were prepared from Al, n-butanol andethyl acetoacetate by the sol-gel route and the N2 supercritical extraction drying technique.Optical experiment results showed that for alumina aerogels after annealing at 500℃ for 4 h pneintense blue emission band appeared in the wavelength range of 400 to 800 nm, and the opticalabsorption edge moved from 8.0 eV of coarse grain Al2O3 to 3~5 eV and with increasing theannealing temperature, this luminescence band presented blue shifts. The reasons of these newoptical properties are discussed in detail.

  15. PPL catalyzed four-component PASE synthesis of 5-monosubstituted barbiturates: Structure and pharmacological properties.

    Bihani, Manisha; Bora, Pranjal P; Verma, Alakesh K; Baruah, Reshita; Boruah, Hari Prasanna Deka; Bez, Ghanashyam


    Enzymatic four-component reactions are very rare although three-component enzymatic promiscuous reactions are widely reported. Herein, we report an efficient PASE protocol for the synthesis of potentially lipophilic zwitterionic 5-monosubstituted barbiturates by four component reaction of mixture of ethyl acetoacetate, hydrazine hydrate, aldehyde and barbituric acid in ethanol at room temperature. Seven different lipases were screened for their promiscuous activity towards the synthesis of 5-monosubstituted barbiturates and the lipase from porcine pancreas (PPL) found to give optimum efficiency. The zwitterionic 5-monosubstituted barbiturates with pyrazolyl ring showed promising pharmacological activity upon screening for antibacterial and apoptotic properties.

  16. Acetoacetylation of Hydroxyethyl Cellulose

    陈晓锋; 高彦芳; 杜奕; 刘德山


    The acetoacetyl group can be used to improve superabsorbent resins since it is more active than the hydroxyethyl group. The acetoacetyl group can be introduced into the side group of hydroxyethyl cellulose (HEC) to activate HEC using the ester exchange reaction between HEC and ethyl acetoacetate (EAA) to improve HEC grafting. This paper discusses the main factors affecting the reaction, such as the amount of EAA and catalyzer, the reaction temperature, and the reaction time. The acetoacetyl group was successfully introduced into HEC. Within specified ranges, increasing the amount of EAA, the reaction temperature and the reaction time will increase the acetoacetylation.

  17. A microwave-catalyzed rapid, efficient and ecofriendly synthesis of substituted pyrazol-5-ones



    Full Text Available A series of 1-(3,4-dihydro-3-oxo-2H-1,4-benzoxazine-2-carbonyl-3-methyl-4-(substituted phenylhydrazono-2-pyrazolin-5-ones have been synthesized by the reaction of 2H-3,4-dihydro-3-oxo-1,4-benzoxazine-2-carboxylic acid hydrazide with substituted acetoacetic ester derivatives using acetic acid as solvent under microwave irradiation (MWI, as well as by conventional methods. The reaction rate is enhanced tremendously and the yields are improved under MWI as compared to conventional methods.

  18. One-pot solvent free synthesis of dihydropyrimidinones using calcined Mg/Fe hydrotalcite catalyst

    Vijay V Dabholkar


    Full Text Available The Mg/Fe = 3 hydrotalcite as reusable solid catalyst was found to be an excellent heterogeneous base catalyst for the synthesis of 3,4-dihydropyrimidinones/thiones, a multicomponent reaction using substituted aromatic aldehyde, ethyl acetoacetate and urea/thiourea. The work has been carried out under thermal solvent free conditions. Mg/Fe = 3 hydrotalcite heterogeneous solid catalyst offers simple means of recovery and the isolated catalyst was reused for number of times without significant loss of catalytic activity.

  19. Metabolic effect of islet B-cell function in insulin-treated diabetes

    Gjessing, H J; Matzen, L E; Iversen, S;


    We studied the relationship between endogenous insulin secretion and fasting levels of plasma free fatty acids (FFA), plasma acetoacetate plus plasma 3-hydroxybutyrate (total ketone bodies), blood glucose, and HbA1 in 132 diabetic outpatients treated with conventional insulin regimens. Patients.......60 nmol/l, and one group with C-peptide stimulation greater than 0.60 nmol/l. According to clinical criteria the prevalence of insulin-dependent diabetes mellitus was approximately 90% in patients with C-peptide stimulation less than 0.32 nmol/l, approximately 25% in patients with C-peptide stimulation...

  20. DFT study on mechanism of the classical Biginelli reaction

    Jin Guang Ma; Ji Ming Zhang; Hai Hui Jiang; Wan Yong Ma; Jian Hua Zhou


    The condensation of benzaldehyde, urea, and ethyl acetoacetate according to the procedure described by Biginelli was investigated at the B3LYP/6-31G(d), B3LYP/6-31+G(d,p), and B3LYP/6-311+G(3df,2p)//B3LYP/6-31+G(d,p) levels to explore the reaction mechanism. According to the mechanism proposed by Kappe, structures of five intermediates were optimized and four transition states were found. The calculation results proved that the mechanism proposed by Kappe is right.

  1. [The establishment of the MPEL for carboxylic acid derivatives in the air of a work area by using mathematical models].

    Gurova, A I; Smoliar, N Ia; Drozhzhina, N A


    Reports of MACs for carboxylic acid derivatives as sodium methoxyacetate (SMOA), methyl chlorpropionate (MCP) and acetoacetic ether (AAE). SMOA shows low toxicity, no cumulation and specific action MCP demonstrates moderate toxicity, low cumulation no specific action in concentrations close to Limac. AAE presents toxicity low in oral usage and moderate when inhaled, low cumulation, no irritation and sensitization. Tentative safe level of injury and MAC estimated for the aforesaid substances together with toxicity parameters compared with the substances studied before allowed the authors to set the MAC of SMOA--10, MCP--2, AAE--10 mg/m3.

  2. Synthesis of New dihydropyrimidinones catalysed by dicationic ionic liquid

    Dhanaji V Jawale; Umesh R Pratap; Aparna A Mulay; Jyotirling R Mali; Ramrao A Mane


    A convenient multi step synthetic protocol for new dihydropyrimidinones bearing quinolynyl methoxy phenyl moiety has been developed from 2-chloro-3-formyl quinolines. The last step is one-pot Biginelli reaction of multicomponents, 4-((2-chloroquinolin-3-yl) methoxy) benzaldehydes, ethyl acetoacetate and urea mediated and catalysed by dicationic ionic liquid (3-methyl-1-[3-(methyl-1H-imidazolium-1-yl) propyl]-1H-imidazolium dibromide (C3 [min]2 2 [Br−] )). Simple work-up procedures and moderate to good yields of the pyrimidinones and the intermediates are the merits of the route.

  3. Potential antibacterial activity of coumarin and coumarin-3-acetic acid derivatives.

    Chattha, Fauzia Anjum; Munawar, Munawar Ali; Nisa, Mehrun; Ashraf, Mohammad; Kousar, Samina; Arshad, Shafia


    Coumarin and coumarin-3-acetic acid derivatives were synthesized by reacting phenols with malic acid, ethyl acetoacetate and ethyl acetylsuccinate in appropriate reaction conditions. All synthesized compounds were subjected to test for their antimicrobial activities against variety of gram positive (Bacillus subtilis, Staphylococcus aureus) and gram negative bacterial stains (Shigella sonnei, Escherichia coli) by agar dilution method. Several of them exhibited appreciable good antibacterial activity against the different strains of gram positive and gram negative bacteria. These findings suggest a great potential of these compounds for screening and use as antibacterial agents for further studies with a battery of bacteria.

  4. Reactions of CH-acids with α,β-unsaturated aldehydes in ionic liquids

    Kryshtal, G. V.; Zhdankina, G. M.; Astakhova, Irina Kira


    Metal carbonate-catalyzed reactions of CH-acids (diethyl malonate, ethyl acetoacetate, ethyl cyanoacetate, and ethyl 2-acetyl- and 2-ethoxycarbonyl-5,9- dimethyldeca-4,8-dienoates) with α,β-unsaturated aldehydes (acrolein, crotonaldehyde, citral) were studied in an ionic liquid, 1-butyl-3......- methylimidazolium hexafluorophosphate [bmim][PF 6], and in a 1-butyl-3-methylimidazolium bromide ([bmim][Br]) - benzene system. The reactions with acrolein and crotonaldehyde afforded Michael addition products, those with citral resulted in Knoevenagel addition products. Sonication increased the yields...

  5. Unsaturated b-ketoesters and their Ni(II, Cu(II and Zn(II complexes



    Full Text Available A new series of b-ketoesters in which the keto group is attached to the olefinic linkage were synthesized by the reaction of methyl acetoacetate and aromatic aldehydes under specified conditions. The existence of these compounds predominantly in the intramolecularly hydrogen bonded enol form was well demonstrated from their IR, 1H-NMR and mass spectral data. Details on the formation of their [ML2] complexes with Ni(II, Cu(II and Zn(II and the nature of the bonding are discussed on the basis of analytical and spectral data.

  6. Identification of the phosphorylation site of the histidine kinase of E. coli AtoS-AtoC two-component system

    Filippou, P.S.; Kasemian, L. D.; Panagiotidis, C A; Kyriakidis, D A


    Abstract Journal URL: The sensor histidine kinase AtoS together with AtoC/Az constitute a two-component signal transduction system (TCS) in E. coli, involved in the regulation of the atoDAEB operon. Upon activation by acetoacetate, AtoS autophosphorylates and subsequently phosphorylates AtoC which is essential for the transcriptional regulation of the atoDAEB operon, the products of which are involved in the catabolism of sho...

  7. Highly efficient solvent-free synthesis of pyranopyrazoles by a Brønsted-acidic ionic liquid as a green and reusable catalyst

    Javad Ebrahimi; Ali Mohammadi; Vahid Pakjoo; Esan Bahramzade; Amir Habibi


    A simple, green and efficient protocol for synthesis of dihydropyrano[2,3-c]pyrazole derivatives is developed by a four component reaction of various benzaldehydes, ethyl acetoacetate, hydrazine hydrate and malononitrile in the presence of 3-methyl-1-(4-sulphonic acid)butylimidazolium hydrogen sulphate [(CH2)4SO3HMIM][HSO4], an acidic ionic liquid and as a catalyst, under solvent-free conditions. The key advantages of this process are high yields, shorter reaction times, easy work-up, purification of products by non-chromatographic method and the reusability of the catalyst.

  8. Interrelations between C4 Ketogenesis, C5 Ketogenesis, and Anaplerosis in the Perfused Rat Liver*

    Deng, Shuang; Zhang, Guo-Fang; Kasumov, Takhar; Roe, Charles R.; Brunengraber, Henri


    We investigated the interrelations between C4 ketogenesis (production of β-hydroxybutyrate + acetoacetate), C5 ketogenesis (production of β-hydroxypentanoate + β-ketopentanoate), and anaplerosis in isolated rat livers perfused with 13C-labeled octanoate, heptanoate, or propionate. Mass isotopomer analysis of C4 and C5 ketone bodies and of related acyl-CoA esters reveal that C4 and C5 ketogenesis share the same pool of acetyl-CoA. Although the uptake of octanoate and heptanoate by the liver ar...

  9. A Solvent-Free Claisen Condensation Reaction for the Organic Laboratory

    Esteb, John J.; Stockton, Matthew B.


    An experiment involving the Claisen condensation reaction for a first-year organic chemistry laboratory is presented. Claisen condensations are routinely covered in organic textbooks but owing to the long reaction times required to reach equilibrium in solution they are seldom explored in the undergraduate teaching laboratory. In this experiment, potassium tert-butoxide and ethyl phenylacetate are heated to 100 °C for 30 minutes under solvent-free conditions to produce 2,4-diphenyl acetoacetate in 80% yield. The solvent-free nature of this procedure greatly reduces the quantity of waste generated by students relative to typical carbonyl condensation experiments.


    V. Khatri, K. Sharma, V. Sareen, D. Shinde and S. Sareen


    Full Text Available Some new 4-[(substituted-benzothiazol-2-ylhydrazono]-2-(substituted-phenyl-5- methyl/ethoxy-2,4-dihydro-pyrazol-3-one(4 have been synthesized by reacting substituted 2- amino benzothiazol (1 with acetoacetic ester and malonic ester (2. 2-[(substituted-benzothiazol- 2-ylhydrazono]-3-oxo-butyric acid ethyl ester and 2-[(substituted-benzothiazol-2- ylhydrazono]- malonic acid diethyl ester (3 react with different hydrazines to give the title compounds(4. These compounds are evaluated for their antifungal and insecticidal activity. The structures of all these compounds have been confirmed by IR, 1H NMR, mass spectra and elemental analysis data.

  11. Preparation of fructone catalyzed by water-soluble Br(φ)nsted acid ionic liquids

    Yuan Yuan Wang; Rong Wang; Liang Chun Wu; Li Yi Dai


    Fructone (2-methyl-2-ethylacetoacetate-1,3-dioxolane), a flavouring material, has been synthesized from ethyl acetoacetate and glycol using five water-soluble Br(φ)nsted acid ionic liquids as catalysts for the first time. The used Br(φ)nsted acid ionic liquids include [Hmim]Tfa, [Hmim]Tsa, [Hmim]BF4, [Bmim]HSO4, [Bmim]H2PO4, and [Hmim]BF4 showed the highest catalytic activity for the preparation of fructone. After reaction, the product could be isolated from the reaction system automatically, and the ionic liquid could be directly reused without dehydration.

  12. Microwave assisted synthesis of novel Hantzsch 1,4-dihydropyridines, acridine-l,8-diones and polyhydroquinolines bearing the tetrazolo [1,5-a]quinoline moiety and their antimicrobial activity assess

    Niraj K. Ladani; Divyesh C. Mungra; Manish P. Patel; Ranjan G. Patel


    Microwave assisted efficient Hantzsch reaction via four-component coupling reactions of tetrazolo [l,5-a]quinoline-4-carbal-dehyde, dimedone/cyclohexane-l,3-dione, ethyl/methyl acetoacetate and ammonium acetate was described as the preparation of tetrazolo [l,5-a]quinoline based 1,4-dihydropyridines, acridine-l,8-diones and polyhydroquinolines. The process presented here is simple, rapid, environmentally welcoming and high yielding. All the derivatives were subjected to an in vitro antimicrobial screening against a representative panel of bacteria and fungi and results worth further investigations.

  13. A gas chromatography-mass spectrometry method for the determination of delta-aminolevulinic acid in plant leaves.

    Hijaz, Faraj; Killiny, Nabil


    Delta-aminolevulinic (δ-ALA) acid is an important intermediate for tetrapyrroles biosynthesis and it has recently received great attention in plant physiology and human toxicology. However, the colorimetric method which is the most common method for determination of δ-ALA is time consuming and is not specific. In this study, a method for determination of δ-ALA in plant tissues was developed based on the trimethylsilyl (TMS) derivative of the pyrrole formed from the reaction of δ-ALA with ethyl acetoacetate via Knorr condensation. The δ-ALA in the HCl extract was reacted with ethyl acetoacetate to form a pyrrole. Then, the pyrrole compound was extracted using ethyl acetate and the solvent was evaporated to dryness. The dried sample was derivatized to its TMS ester and analyzed using GC-MS. The concentration of δ-ALA in citrus leaves incubated with levulinic acid was also determined by the conventional colorimetric method. The linear range was 10-200ppm in the full scan mode and 0.1-20ppm in the selected ion monitoring (SIM). The limit of detection was 6ppm in the full scan and 0.05ppm in SIM mode, representing a four-fold increase in sensitivity compared to the colorimetric method. The GC-MS method developed in this study is simple, accurate, sensitive, and could also be used to measure δ-ALA in other biological samples.

  14. A modified pathway for the production of acetone in Escherichia coli.

    May, Antje; Fischer, Ralf-Jörg; Maria Thum, Simone; Schaffer, Steffen; Verseck, Stefan; Dürre, Peter; Bahl, Hubert


    A modified synthetic acetone operon was constructed. It consists of two genes from Clostridium acetobutylicum (thlA coding for thiolase and adc coding for acetoacetate decarboxylase) and one from Bacillus subtilis or Haemophilus influenzae (teII(srf) or ybgC, respectively, for thioesterase). Expression of this operon in Escherichia coli resulted in the production of acetone starting from the common metabolite acetyl-CoA via acetoacetyl-CoA and acetoacetate. The thioesterases do not need a CoA acceptor for acetoacetyl-CoA hydrolysis. Thus, in contrast to the classic acetone pathway of Clostridium acetobutylicum and related microorganisms which employ a CoA transferase, the new pathway is acetate independent. The genetic background of the host strains was crucial. Only E. coli strains HB101 and WL3 were able to produce acetone via the modified plasmid based pathway, up to 64mM and 42mM in 5-ml cultures, respectively. Using glucose fed-batch cultures the concentration could be increased up to 122mM acetone with HB101 carrying the recombinant plasmid pUC19ayt (thioesterase from H. influenzae). The formation of acetone led to a decreased acetate production by E. coli.

  15. Changes of NF-KB activity in colon carcinoma cells treated with different crude extracts of abrotani herba

    Feng Pan; Yuying Chen; Li Yang; Zhiheng Bian; Houjie Liang


    Objective: To study changes of NF-KB activity in colon carcinoma cell lines treated with different crude extracts of abrotani herba obtained through solvent extraction methods.Methods: Crude extracts of abrotani herba were extracted with ligarine, chloroform, acetoacetate and n-butanol in separating funnel.Exposure concentration of crude extracts were obtained through detecting viability of HT-29 cells by MTT.Then HT-29 cells and Lovo cells were treated with different crude extracts respectively.Changes of NF-KB activity in HT-29 cells and Lovo cells using different crude extracts were observed by EMSA.Results: Successfully extracted different crude extracts of abrotani herba and called them ligarine extract, chloroform extract,acetoacetate extract, n-butanol extract and remaining extract for short.NF-KB activity was significantly inhibited in HT-29 cells treated with chloroform extract, there were no significant differences in other groups compared with the control.The same change of NF-KB activity was observed in Lovo calls using different crude extracts of abrotani herba.Conclusion: NF-KB activity can be inhibited in colon carcinoma HT-29 calls and Lovo cells treated with chloroform extract obtained from abrotani herba through the method of solvent extraction.

  16. Tor-Sch9 deficiency activates catabolism of the ketone body-like acetic acid to promote trehalose accumulation and longevity.

    Hu, Jia; Wei, Min; Mirzaei, Hamed; Madia, Federica; Mirisola, Mario; Amparo, Camille; Chagoury, Shawna; Kennedy, Brian; Longo, Valter D


    In mammals, extended periods of fasting leads to the accumulation of blood ketone bodies including acetoacetate. Here we show that similar to the conversion of leucine to acetoacetate in fasting mammals, starvation conditions induced ketone body-like acetic acid generation from leucine in S. cerevisiae. Whereas wild-type and ras2Δ cells accumulated acetic acid, long-lived tor1Δ and sch9Δ mutants rapidly depleted it through a mitochondrial acetate CoA transferase-dependent mechanism, which was essential for lifespan extension. The sch9Δ-dependent utilization of acetic acid also required coenzyme Q biosynthetic genes and promoted the accumulation of intracellular trehalose. These results indicate that Tor-Sch9 deficiency extends longevity by switching cells to an alternative metabolic mode, in which acetic acid can be utilized for the storage of stress resistance carbon sources. These effects are reminiscent of those described for ketone bodies in fasting mammals and raise the possibility that the lifespan extension caused by Tor-S6K inhibition may also involve analogous metabolic changes in higher eukaryotes.

  17. Application of (1)H NMR-based serum metabolomic studies for monitoring female patients with rheumatoid arthritis.

    Zabek, Adam; Swierkot, Jerzy; Malak, Anna; Zawadzka, Iga; Deja, Stanisław; Bogunia-Kubik, Katarzyna; Mlynarz, Piotr


    Rheumatoid arthritis is a chronic autoimmune-based inflammatory disease that leads to progressive joint degeneration, disability, and an increased risk of cardiovascular complications, which is the main cause of mortality in this population of patients. Although several biomarkers are routinely used in the management of rheumatoid arthritis, there is a high demand for novel biomarkers to further improve the early diagnosis of rheumatoid arthritis, stratification of patients, and the prediction of a better response to a specific therapy. In this study, the metabolomics approach was used to provide relevant biomarkers to improve diagnostic accuracy, define prognosis and predict and monitor treatment efficacy. The results indicated that twelve metabolites were important for the discrimination of healthy control and rheumatoid arthritis. Notably, valine, isoleucine, lactate, alanine, creatinine, GPC  APC and histidine relative levels were lower in rheumatoid arthritis, whereas 3-hydroxyisobutyrate, acetate, NAC, acetoacetate and acetone relative levels were higher. Simultaneously, the analysis of the concentration of metabolites in rheumatoid arthritis and 3 months after induction treatment revealed that L1, 3-hydroxyisobutyrate, lysine, L5, acetoacetate, creatine, GPC+APC, histidine and phenylalanine were elevated in RA, whereas leucine, acetate, betaine and formate were lower. Additionally, metabolomics tools were employed to discriminate between patients with different IL-17A genotypes. Metabolomics may provide relevant biomarkers to improve diagnostic accuracy, define prognosis and predict and monitor treatment efficacy in rheumatoid arthritis.

  18. Metabolic Rewiring by Oncogenic BRAF V600E Links Ketogenesis Pathway to BRAF-MEK1 Signaling.

    Kang, Hee-Bum; Fan, Jun; Lin, Ruiting; Elf, Shannon; Ji, Quanjiang; Zhao, Liang; Jin, Lingtao; Seo, Jae Ho; Shan, Changliang; Arbiser, Jack L; Cohen, Cynthia; Brat, Daniel; Miziorko, Henry M; Kim, Eunhee; Abdel-Wahab, Omar; Merghoub, Taha; Fröhling, Stefan; Scholl, Claudia; Tamayo, Pablo; Barbie, David A; Zhou, Lu; Pollack, Brian P; Fisher, Kevin; Kudchadkar, Ragini R; Lawson, David H; Sica, Gabriel; Rossi, Michael; Lonial, Sagar; Khoury, Hanna J; Khuri, Fadlo R; Lee, Benjamin H; Boggon, Titus J; He, Chuan; Kang, Sumin; Chen, Jing


    Many human cancers share similar metabolic alterations, including the Warburg effect. However, it remains unclear whether oncogene-specific metabolic alterations are required for tumor development. Here we demonstrate a "synthetic lethal" interaction between oncogenic BRAF V600E and a ketogenic enzyme 3-hydroxy-3-methylglutaryl-CoA lyase (HMGCL). HMGCL expression is upregulated in BRAF V600E-expressing human primary melanoma and hairy cell leukemia cells. Suppression of HMGCL specifically attenuates proliferation and tumor growth potential of human melanoma cells expressing BRAF V600E. Mechanistically, active BRAF upregulates HMGCL through an octamer transcription factor Oct-1, leading to increased intracellular levels of HMGCL product, acetoacetate, which selectively enhances binding of BRAF V600E but not BRAF wild-type to MEK1 in V600E-positive cancer cells to promote activation of MEK-ERK signaling. These findings reveal a mutation-specific mechanism by which oncogenic BRAF V600E "rewires" metabolic and cell signaling networks and signals through the Oct-1-HMGCL-acetoacetate axis to selectively promote BRAF V600E-dependent tumor development.

  19. Comparative acute effects of leptin and insulin on gluconeogenesis and ketogenesis in perfused rat liver.

    Borba-Murad, Glaucia Regina; Mario, Erica Guilhen; Bassoli, Bruna Kempfer; Bazotte, Roberto Barbosa; de Souza, Helenir Medri


    The acute effects of physiological levels of leptin (10 ng ml(-1)) and insulin (20 microU ml(-1)) on hepatic gluconeogenesis and ketogenesis were compared. Leptin or insulin alone decreased (p<0.05) the activation of hepatic glucose, L-lactate and urea production from L-alanine. However, the hepatic glucose production was not modified if leptin was combined with insulin. These results indicated that both, i.e. leptin and insulin, could promote a non-additive reduction in the rate of catabolism of L-alanine. However, in contrast with insulin (p<0.05), leptin did not inhibit the activation of hepatic glucose production from pyruvate or glycerol. On the other hand, activation of hepatic production of acetoacetate and beta-hydroxybutyrate from octanoate was not affected by leptin or insulin. Thus, our data demonstrate that the acute effect of leptin on hepatic metabolism was partially similar to insulin (activation of glucose production from L-alanine and activation of acetoacetate or beta-hydroxybutyrate production from octanoate) and partially different from insulin (activation of glucose production from pyruvate or glycerol).

  20. Mangiferin supplementation improves serum lipid profiles in overweight patients with hyperlipidemia: a double-blind randomized controlled trial.

    Na, Lixin; Zhang, Qiao; Jiang, Shuo; Du, Shanshan; Zhang, Wei; Li, Ying; Sun, Changhao; Niu, Yucun


    Our previous studies have shown that mangiferin decreased serum triglycerides and free fatty acids (FFAs) by increasing FFAs oxidation in both animal and cell experiments. This study sought to evaluate the effects of mangiferin on serum lipid profiles in overweight patients with hyperlipidemia. Overweight patients with hyperlipidemia (serum triglyceride ≥ 1.70 mmol/L, and total cholesterol ≥ 5.2 mmol/L) were included in this double-blind randomized controlled trial. Participants were randomly allocated to groups, either receiving mangiferin (150 mg/day) or identical placebo for 12 weeks. The lipid profile and serum levels of mangiferin, glucose, L-carnitine, β-hydroxybutyrate, and acetoacetate were determined at baseline and 12 weeks. A total of 97 participants completed the trial. Compared with the placebo control, mangiferin supplementation significantly decreased the serum levels of triglycerides and FFAs, and insulin resistance index. Mangiferin supplementation also significantly increased the serum levels of mangiferin, high-density lipoprotein cholesterol, L-carnitine, β-hydroxybutyrate, and acetoacetate, and increased lipoprotein lipase activity. However, there were no differences in the serum levels of total cholesterol, low-density lipoprotein cholesterol, serum glucose, and insulin between groups. Mangiferin supplementation could improve serum lipid profiles by reducing serum triglycerides and FFAs in overweight patients with hyperlipidemia, partly due to the promotion of FFAs oxidation.

  1. Profile of organic acid concentrations in the digestive gland and hemolymph of Biomphalaria glabrata under estivation

    José Clecildo Barreto Bezerra


    Full Text Available Using high performance liquid chromatography (HPLC analysis it was possible to determine simultaneously the concentration of organic acids (pyruvate, lactate, succinate, fumarate, malate, acetate, propionate, acetoacetate, and ß-hydroxybutyrate in the digestive gland and the extracellular concentration of these same acids in the hemolymph of estivating Biomphalaria glabrata, the intermediate host of Schistosoma mansoni. After a 7 day period of estivation, there was a significant increase in the tissue levels of lactate, succinate, malate and acetate compared to non-estivating snails. After 14 days of estivation, the levels of lactate and acetate were also significantly elevated. The hemolymph concentrations of pyruvate and acetate increased significantly after 7 days and acetate concentrations continued to be significantly increased up to 14 days of estivation. The other organic acids studied, such as ketone body acetoacetate and ß-hydroxybutyrate or the volatile acid propionate, did not accumulate. Their tissue concentrations, however, increased on the 7th day of estivation and reached normal levels within two weeks of estivation for some of them. One should take into consideration how the reduction in metabolism can be handled under aerobic conditions, and what role anaerobic pathways may play in both energy formation and redox balance processes.

  2. Lactate as a cerebral metabolic fuel for glucose-6-phosphatase deficient children.

    Fernandes, J; Berger, R; Smit, G P


    The main substrates for brain energy metabolism were measured in blood samples taken from the carotid artery and the internal jugular bulb of four children with glycogen storage disease caused by deficiency of glucose-6-phosphatase. Multiple paired arterial and venous blood samples were analyzed for glucose, lactate, pyruvate, D-beta-hydroxybutyrate, acetoacetate, glycerol and O2, and the arteriovenous differences of the concentrations were calculated. In the first three patients the substrates were measured in two successive conditions with lower and higher glucose-intake, respectively, inducing reciprocally higher and lower concentrations of blood lactate. In the fourth patient medium chain triglycerides were administered simultaneously with the glucose-containing gastric drip feeding. Lactate appeared to be taken up significantly. It consumed, if completely oxidized, between 40-50% of the total O2 uptake in most cases. Only once in one patient the uptake of lactate switched to its release, when the blood lactate level decreased to normal. D-beta-hydroxybutyrate and acetoacetate arteriovenous (A-V) differences were small to negligible and these ketone bodies, therefore, did not contribute substantially to the brain's energy expenditure. Glycerol was not metabolized by the brain. Lactate thus appeared to be the second brain fuel next to glucose. It may protect the brain against fuel depletion in case of hypoglycemia.

  3. Study on Screening Valid Target of Anti -inflammatory and Analgesic Effets of Calystegia Soldanella%孝扇草抗炎镇痛有效部位的筛选研究

    谭丽; 沈清; 黄真


    Objective: To separate and screen the valid target of anti - inflammatory and analgesic effects of Calystegia soldanella . Methods: Systematic solvent method is used to separate the ethanol extract of Calystegia soldanella ,ear edema induced by dimethylbenzene in mice was used to screen the valid target of the anti - inflammatory effects,mice hot -plate procedures and mice wthithing induced by acetic acid were used to screen the valid target of the analgesic effects. Results :The inhibition ratio of swelling induced by dimethylbenzene in mice of the ethanol - extracted acetoacetate - abstraction group of Calystegia soldanella is 50.53% (P < 0.05). The acetoacetate - abstraction group and the rest group could significantly prolong the mouse hot - plate - induced pain of the pain threshold of the mice( P <0.05 ,P <0.05). The ethanol - extracted acetoacetate - abstraction group and the n - buOH - abstraction group can significantly decrease ? acetic acid -induced writhing response in mice(P < 0.05, P < 0.05). Conclusion-.The ethanol -extracted acetoacetate -abstraction group of Calystegia soldanella is most propably the valid target of anti - inflammatory and analgesic.%目的:分离并筛选孝扇草抗炎镇痛作用的有效部位.方法:采用系统溶剂法对孝扇草醇提物进行初步分离;采用二甲苯致小鼠耳廓肿胀实验,筛选孝扇草抗炎作用有效部位;采用热板法和醋酸致小鼠扭体法实验,筛选其镇痛作用有效部位.结果:孝扇草醇提乙酸乙酯萃取部位对二甲苯致小鼠耳廓肿胀的抑制率为50.53%,与空白对照组比较有显著性差异(P<0.05);孝扇草醇提乙酸乙酯萃取部位和各溶剂萃取剩余部位均能显著提高小鼠痛阈值,与空白对照组比较有显著性差异(P <0.05,P<0.05);孝扇草醇提乙酸乙酯和正丁醇萃取部位均能明显减少醋酸致小鼠扭体反应次数,与空白组比较有显著性差异(P<0.05,P<0.05).结论:孝扇草醇提

  4. Multicomponent Synthesis and Evaluation of New 1,2,3-Triazole Derivatives of Dihydropyrimidinones as Acidic Corrosion Inhibitors for Steel.

    González-Olvera, Rodrigo; Román-Rodríguez, Viridiana; Negrón-Silva, Guillermo E; Espinoza-Vázquez, Araceli; Rodríguez-Gómez, Francisco Javier; Santillan, Rosa


    An efficient one-pot synthesis of 1,2,3-triazole derivatives of dihydropyrimidinones has been developed using two multicomponent reactions. The aldehyde-1,2,3-triazoles were obtained in good yields from in situ-generated organic azides and O-propargylbenzaldehyde. The target heterocycles were synthesized through the Biginelli reaction in which the aldehyde-1,2,3-triazoles reacted with ethyl acetoacetate and urea in the presence of Ce(OTf)₃ as the catalyst. The corrosion inhibition of steel grade API 5 L X52 in 1 M HCl by the synthesized compounds was investigated using the electrochemical impedance spectroscopy technique. The measurements revealed that these heterocycles are promising candidates to inhibit acidic corrosion of steel.

  5. ß-Phenylethylamine as a novel nutrient treatment to reduce bacterial contamination due to Escherichia coli O157:H7 on beef meat.

    Lynnes, Ty; Horne, S M; Prüß, B M


    Bacterial infection by Escherichia coli O157:H7 through the consumption of beef meat or meat products is an ongoing problem, in part because bacteria develop resistances towards chemicals aimed at killing them. In an approach that uses bacterial nutrients to manipulate bacteria into behaviors or cellular phenotypes less harmful to humans, we screened a library of 95 carbon and 95 nitrogen sources for their effect on E. coli growth, cell division, and biofilm formation. In the initial screening experiment using the Phenotype MicroArray(TM) technology from BioLog (Hayward, CA), we narrowed the 190 starting nutrients down to eight which were consecutively tested as supplements in liquid beef broth medium. Acetoacetic acid (AAA) and ß-phenylethylamine (PEA) performed best in this experiment. On beef meat pieces, PEA reduced the bacterial cell count by 90% after incubation of the PEA treated and E. coli contaminated meat pieces at 10°C for one week.

  6. Synthesis of Some Pyrazolone Derivatives and Evaluation of its Antibacterial and Cytotoxic Activity

    Rishiram Prajuli


    Full Text Available A series of novel pyrazolone derivative were synthesized by two different schemes (scheme-1 by the reaction of phenyl hydrazine and ethyl acetoacetate with substituted benzaldehydes PYR-1 to PYR-4 and (by the reaction of synthesized chalcone with phenyl hydrazine PYR-5 and characterised with its physical parameters (M.P, colour, %yield, solubility etc.. The entire synthesized compound was tested for their antimicrobial activity against Gram-positive and Gram-negative strains of bacteria and brimeshrimp bioassay was conducted for evaluation of cytotoxic activity The Investigation of antimicrobial screening data revealed that most of the tested compounds showed moderate to good antimicrobial activity. And cytotoxicity activity of compounds was also found to be satisfactory.

  7. Evaluation of the Accuracy of Capillary Hydroxybutyrate Measurement Compared with Other Measurements in the Diagnosis of Diabetic Ketoacidosis: A Systematic Review.

    Brooke, Joanne; Stiell, Marlon; Ojo, Omorogieva


    A complication of diabetes is diabetic ketoacidosis (DKA), which if left untreated is a life threatening condition. Prompt and accurate diagnosis of DKA is required for the commencement of life saving interventions. Measurements of ketone bodies in DKA have usually been through nitroprusside urine acetoacetate testing. The aim of this systematic review was to examine whether capillary β-hydroxybutyrate (β-OHB) testing is more accurate compared to other diagnostic methods of DKA. The following electronic databases were searched: EBSCO Host, MEDLINE, PSYCHInfo, CINAHL and Science Direct for publications from 1 January 2005 and up to and including 1 January 2016. Inclusion criteria were: Adults 18 years and over and known type 1 or type 2 diabetes. Retrospective and prospective observation studies were included. A total of nine studies met the inclusion criteria. Capillary β-OHB was found to have high sensitivity, specificity, positive predictive value and negative predictive value in identifying DKA compared to urinary ketone testing.

  8. Synthesis of functionalized pyrazolopyran derivatives: comparison of two-step vs. one-step vs. microwave-assisted protocol and X-ray crystallographic analysis of 6-amino-1,4-dihydro-3-methyl-4-phenylpyrano[2,3-c]pyrazole-5-carbonitrile

    Paritosh Shukla; Ashok Sharma; Sumati Anthal; Rajni Kant


    A library of pyrazolopyran heterocycles was synthesized first by the traditional heating techniques using two-step and one-step procedures and then by microwave-assisted (MWA) multicomponent condensation of ethyl acetoacetate, hydrazine, malonodinitrile and a variety of substituted aldehydes. A comparison of the foregoing methods was first done based on the yields and then based on the time taken for the completion of the reactions. It was found that although the traditional methods gave slightly better yields, the MWA syntheses lead to substantial reduction in reaction timings. The title compound crystallizes in the triclinic crystal system with space group P–1. The crystal structure as elucidated by X-ray diffraction methods shows the presence of different intermolecular interactions, and the nature and energetics associated with these interactions have been characterized using PIXEL software.

  9. Schiff bases of N-(2-aminoethyl)-3-aminopropyltrimethoxysilane and its silatranes: Synthesis and characterization

    Gurjaspreet Singh; Amandeep Saroa; Sadhika Khullar; Sanjay K Mandal


    This paper aims at the introduction of azomethine group by the condensation reaction of N-(2-aminoethyl)-3-aminopropyltrimethoxysilane with different compounds containing carbonyl group such as 2’-hydroxyacetophenone, salicylaldehyde, pyrrole-2-carboxaldehyde, acetylacetone and ethyl acetoacetate. Further, transesterification reaction of these Schiff base modified silanes with triethanolamine as a tripodal ligand leads to the synthesis of corresponding silatranes 1–5 bearing Schiff base functionalized long chain in the axial position. All the synthesized compounds are characterized by spectroscopic methods, elemental analysis and mass spectrometry. The authentication of Schiff base modified silatranes is scrutinized by single X-ray crystal structure of silatrane 1. The thermal stability of the five silatranes is studied by thermo-gravimetric analysis (TGA).

  10. A facile approach for the construction of the oxetane ring from 5α-acyloxy-△4(20)-taxoids

    LIU, Rui-Wu; YIN, Da-Li; GUO, Ji-Yu; LIANG, Xiao-Tian


    An oxetane ring can be constructed from 5α-acyloxy-△4(20)taxoids. Tne facile intramolecular acyl migration from 5- to 20-postion under slightly basic conditions enabled the const-uction of the oxetane ring in a convenient short cut, whereas the acyl migration from 2- to 20-position left the 2-hydroxyl ~~ble to a later benzoylation. An unexpected five-membered 4-O, 20-O sulfite ring was formed in the attempted construction of the oxetane ring with 5α-trifiate as a leaving group. After the construction of the oxetane ring, treatment with strong base LiHMDS and acetyl chloride gave the expected 4-O-acetate while treatment with acetic anhydride and DMAP gave a 4- O-acetoacetate.

  11. Engineered synthetic pathway for isopropanol production in Escherichia coli.

    Hanai, T; Atsumi, S; Liao, J C


    A synthetic pathway was engineered in Escherichia coli to produce isopropanol by expressing various combinations of genes from Clostridium acetobutylicum ATCC 824, E. coli K-12 MG1655, Clostridium beijerinckii NRRL B593, and Thermoanaerobacter brockii HTD4. The strain with the combination of C. acetobutylicum thl (acetyl-coenzyme A [CoA] acetyltransferase), E. coli atoAD (acetoacetyl-CoA transferase), C. acetobutylicum adc (acetoacetate decarboxylase), and C. beijerinckii adh (secondary alcohol dehydrogenase) achieved the highest titer. This strain produced 81.6 mM isopropanol in shake flasks with a yield of 43.5% (mol/mol) in the production phase. To our knowledge, this work is the first to produce isopropanol in E. coli, and the titer exceeded that from the native producers.

  12. Engineering a synthetic pathway in cyanobacteria for isopropanol production directly from carbon dioxide and light.

    Kusakabe, Tamami; Tatsuke, Tsuneyuki; Tsuruno, Keigo; Hirokawa, Yasutaka; Atsumi, Shota; Liao, James C; Hanai, Taizo


    Production of alternate fuels or chemicals directly from solar energy and carbon dioxide using engineered cyanobacteria is an attractive method to reduce petroleum dependency and minimize carbon emissions. Here, we constructed a synthetic pathway composed of acetyl-CoA acetyl transferase (encoded by thl), acetoacetyl-CoA transferase (encoded by atoAD), acetoacetate decarboxylase (encoded by adc) and secondary alcohol dehydrogenase (encoded by adh) in Synechococcus elongatus strain PCC 7942 to produce isopropanol. The enzyme-coding genes, heterogeneously originating from Clostridium acetobutylicum ATCC 824 (thl and adc), Escherichia coli K-12 MG1655 (atoAD) and Clostridium beijerinckii (adh), were integrated into the S. elongatus genome. Under the optimized production conditions, the engineered cyanobacteria produced 26.5 mg/L of isopropanol after 9 days.

  13. Production of isopropanol by metabolically engineered Escherichia coli.

    Jojima, Toru; Inui, Masayuki; Yukawa, Hideaki


    A genetically engineered strain of Escherichia coli JM109 harboring the isopropanol-producing pathway consisting of five genes encoding four enzymes, thiolase, coenzyme A (CoA) transferase, acetoacetate decarboxylase from Clostridium acetobutylicum ATCC 824, and primary-secondary alcohol dehydrogenase from C. beijerinckii NRRL B593, produced up to 227 mM of isopropanol from glucose under aerobic fed-batch culture conditions. Acetate production by the engineered strain was approximately one sixth that produced by a control E. coli strain bearing an expression vector without the clostridial genes. These results demonstrate a functional isopropanol-producing pathway in E. coli and consequently carbon flux from acetyl-CoA directed to isopropanol instead of acetate. This is the first report on isopropanol production by genetically engineered microorganism under aerobic culture conditions.

  14. Synthesis and anti-depressant evaluation of novel pyrazolone derivatives

    Vijay Kumar Merugumolu


    Full Text Available Diazotization of substituted anilines with NaNO2 and concentrated hydrochloric acid at 0ºC gave the diazonium chlorides. Coupling of substituted aryl diazonium chlorides with ethyl acetoacetate in methanol gave ethyl-2-aryl-hydrazono-3-oxobutyrates (2a-h. Reaction of (2a-h with naphthoic carbohydrazide (3 gave the title compounds pyrazolone derivatives (4a-h. The newly synthesized compounds were screened for their in vivo anti-depressant activity by tail suspension test and forced swimming test. Some of the tested compounds 4f, 4g showed very good activity when compared to the standard drug imipramine. The newly synthesized compounds were characterized by physical parameters and the structures were elucidated by spectral data.

  15. Silica-bonded N-propylpiperazine sodium n-propionate as recyclable catalyst for synthesis of 4H-pyran derivatives

    Khodabakhsh Niknam; Nassim Borazjani; Reza Rashidian; Abbas Jamali


    Silica-bonded N-propylpiperazine sodium n-propionate (SBPPSP) was found to act as an efficient solid base for the preparation of a series of 4H-benzo[b]pyran derivatives. SBPPSP was used as a recyclable heterogeneous solid base catalyst for the synthesis of 3,4-dihydropyrano[c]chromenes, 2-amino-4H-pyrans, 1,4-dihydropyrano[2,3-c]pyrazoles, and 2-amino-4H-benzo[e]-chromenes via the condensation reaction of dimedone, ethyl acetoacetate, 3-methyl-1-phenyl-1H-pyrazol-5(4H)-one, andα-naphthol, respectively, with aromatic aldehydes and malononitrile in refluxing aqueous ethanol. The heterogeneous solid base showed similar efficiency when reused in consecutive reac-tions.

  16. Multicomponent Synthesis and Evaluation of New 1,2,3-Triazole Derivatives of Dihydropyrimidinones as Acidic Corrosion Inhibitors for Steel

    Rodrigo González-Olvera


    Full Text Available An efficient one-pot synthesis of 1,2,3-triazole derivatives of dihydropyrimidinones has been developed using two multicomponent reactions. The aldehyde-1,2,3-triazoles were obtained in good yields from in situ-generated organic azides and O-propargylbenzaldehyde. The target heterocycles were synthesized through the Biginelli reaction in which the aldehyde-1,2,3-triazoles reacted with ethyl acetoacetate and urea in the presence of Ce(OTf3 as the catalyst. The corrosion inhibition of steel grade API 5 L X52 in 1 M HCl by the synthesized compounds was investigated using the electrochemical impedance spectroscopy technique. The measurements revealed that these heterocycles are promising candidates to inhibit acidic corrosion of steel.

  17. Biomimetic potential of some methacrylate-based copolymers: a comparative study.

    Zecheru, Teodora; Filmon, Robert; Rusen, Edina; Mărculescu, Bogdan; Zerroukhi, Amar; Cincu, Corneliu; Chappard, Daniel


    Preparation of new biocompatible materials for bone recovery has consistently gained interest in the last few decades. Special attention was given to polymers that contain negatively charged groups, such as phosphate, carboxyl, and sulfonic groups toward calcification. This present paper work demonstrates that other functional groups present also potential application in bone pathology. New copolymers of 2-hydroxyethyl methacrylate with diallyldimethylammonium chloride (DADMAC), glycidyl methacrylate (GlyMA), methacrylic acid (MAA), 2-methacryloyloxymethyl acetoacetate (MOEAA), 2-methacryloyloxyethyltriethylammonium chloride (MOETAC), and tetrahydrofurfuryl methacrylate (THFMA) were obtained. The copolymers were characterized by FTIR, swelling potential, and they were submitted to in vitro tests for calcification and cytotoxicity evaluation. GlyMA and MOETAC-containing copolymers show promising results for further in vivo mineralization tests, as a potential alternative to the classical bone grafts, in bone tissue engineering.

  18. Microwave Assisted Convenient One-Pot Synthesis of Coumarin Derivatives via Pechmann Condensation Catalyzed by FeF3 under Solvent-Free Conditions and Antimicrobial Activities of the Products

    Vahid Vahabi


    Full Text Available A rapid and efficient solvent-free one-pot synthesis of coumarin derivatives by Pechmann condensation reactions of phenols with ethyl acetoacetate using FeF3 as a catalyst under microwave irradiation is described. This one-pot synthesis on a solid inorganic support provides the products in good yields. The newly synthesized compounds were systematically characterized by IR, 1H-NMR, 13C-NMR, MS and elemental CHN analyses. The proposed solvent-free microwave irradiation method using the environmentally friendly catalyst FeF3 offers the unique advantages of high yields, shorter reaction times, easy and quick isolation of the products, excellent chemoselectivity, and a one-pot, green synthesis. The products were screened for antimicrobial activity, and the results showed that the compounds reacted against all the tested bacteria.

  19. Nano Fe{sub 2}O{sub 3,} clinoptilolite and H{sub 3}PW{sub 12}O{sub 40} as efficient catalysts for solvent-free synthesis of 5(4H)-isoxazolone under microwave irradiation conditions

    Fozooni, Samieh, E-mail:, E-mail: [Shahid Bahonar University, Kerman (Iran, Islamic Republic of). Mining Engineering Department. Zarand High Education Center; Hosseinzadeh, Nasrin Gholam; Akhgar, Mohammad Reza [Islamic Azad University, Kerman (Iran, Islamic Republic of). Department of Chemistry; Hamidian, Hooshang [Payame Noor University (PNU), Tehran (Iran, Islamic Republic of). Department of Chemistry


    A quick and solvent-free approach involving the exposure of neat reactants to microwave irradiation in conjunction with the use of clinoptilolite, H{sub 3}PW{sub 12}O{sub 40} and Fe{sub 2}O{sub 3} nanoparticle catalysts is described. In this work, condensation of hydroxylamine hydrochloride, sodium acetate, acetoacetic or benzoyl acetic ethyl ester and appropriate aldehydes by employing catalysts gave 5(4H)-isoxazolone only in one step. Catalyst amount, temperature effects and catalysts reusability were monitored. Among the catalysts, Fe{sub 2}O{sub 3} nanoparticles had better performance than other catalysts from viewpoint of yield and reaction time. The present protocol offers several advantages, such as short reaction time, reasonable yield, mild reaction condition and recycling catalysts with a very easy workup. (author)

  20. Synthesis of Leflunomide%来氟米特的合成

    沈敬山; 王斌; 李剑峰; 雷厉军; 嵇汝运


    以价廉易得的乙酰乙酸乙酯为起始原料,缩合、环合后中间产物3不分离,水解得到68%的5-甲基-4-异FDA9唑甲酸,经氯化、纯化后与对三氟甲基苯胺反应得到来氟米特,两步收率60.8%,总收率41%。操作简便,反应副产物少。%Leflunomide, a new drug for treating rheumatoid arthritis, was synthesized from ethyl acetoacetate and triethyl orthoformate by condensation, cyclization, hydrolysis and chlorination to obtain 5-methyl-4-isoxazolecarboxylic acid chloride which reacted with 4-trifluoromethylaniline to give the desired product in an overall yield of 41%.

  1. Metabolomic investigation of Mytilus galloprovincialis (Lamarck 1819) caged in aquatic environments.

    Fasulo, Salvatore; Iacono, Francesco; Cappello, Tiziana; Corsaro, Carmelo; Maisano, Maria; D'Agata, Alessia; Giannetto, Alessia; De Domenico, Elena; Parrino, Vincenzo; Lo Paro, Giuseppe; Mauceri, Angela


    Environmental metabolomics was applied to assess the metabolic responses in transplanted mussels to environmental pollution. Specimens of Mytilus galloprovincialis, sedentary filter-feeders, were caged in anthropogenic-impacted and reference sites along the Augusta coastline (Sicily, Italy). Chemical analysis revealed increased levels of PAHs in the digestive gland of mussels from the industrial area compared with control, and marked morphological changes were also observed. Digestive gland metabolic profiles, obtained by 1H NMR spectroscopy and analyzed by multivariate statistics, showed changes in metabolites involved in energy metabolism. Specifically, changes in lactate and acetoacetate could indicate increased anaerobic fermentation and alteration in lipid metabolism, respectively, suggesting that the mussels transplanted to the contaminated field site were suffering from adverse environmental condition. The NMR-based environmental metabolomics applied in this study results thus in it being a useful and effective tool for assessing environmental influences on the health status of aquatic organisms.

  2. Silica sulfuric acid: a reusable solid catalyst for one pot synthesis of densely substituted pyrrole-fused isocoumarins under solvent-free conditions

    Sudipta Pathak


    Full Text Available A convenient and efficient methodology for the synthesis of densely substituted pyrrole-fused isocoumarins, which employs solid-supported silica sulfuric acid (SSA as catalyst, has been developed. When the mixture of ninhydrin adducts of acetylacetone/ethyl acetoacetate and primary amines was heated on the solid surface of SSA under solvent-free conditions, the pyrrole-fused isocoumarins were formed in good yields. This synthetic method has several advantages such as the employment of solvent-free reaction conditions without the use of any toxic reagents and metal catalysts, the ease of product isolation, the use of a recyclable catalyst, the low cost, the easy availability of the starting materials, and the excellent yields of products.

  3. Synthesis of Some New Benzimidazole Derivatives of Pharmaceutical Interest

    Fawzia Zakaria El-Ablack


    Full Text Available Reaction of 2-(aminomethylbenzimidazole dihydrochloride (1 with ethyl acetoacetate was studied to give diazepinone-benzimidazole derivative (2, while, treatment of 1 with phenylhydrazono ethylacetoacetate afforded phenylhydrazino diazepinone derivative (3. On the other hand, reaction of 1 with acetyl acetone resulted in the formation of diazepine derivative (4. The reaction of 1 with ethyl cyanoacetate was studied to give 3-aminodiazepinone derivative (5. Also the reaction of 1 with acetophenone and/or benzophenone has been investigated to give the fused imidazolines 6 and 7 respectively, while the reaction of 1 with cyclopentanone gave benzimidazolyl derivative (8. Treatment of 1 with chloroacetyl chloride gave the fused pyrazinone (9. The treatment of 1 with benzoin gave the derivative (10. The structures of the hitherto unknown compounds have been confirmed from analytical and spectral data. The newly synthesized compounds were screened for antibacterial and antifungal activity.

  4. Evaluation of the Accuracy of Capillary Hydroxybutyrate Measurement Compared with Other Measurements in the Diagnosis of Diabetic Ketoacidosis: A Systematic Review

    Joanne Brooke


    Full Text Available A complication of diabetes is diabetic ketoacidosis (DKA, which if left untreated is a life threatening condition. Prompt and accurate diagnosis of DKA is required for the commencement of life saving interventions. Measurements of ketone bodies in DKA have usually been through nitroprusside urine acetoacetate testing. The aim of this systematic review was to examine whether capillary β-hydroxybutyrate (β-OHB testing is more accurate compared to other diagnostic methods of DKA. The following electronic databases were searched: EBSCO Host, MEDLINE, PSYCHInfo, CINAHL and Science Direct for publications from 1 January 2005 and up to and including 1 January 2016. Inclusion criteria were: Adults 18 years and over and known type 1 or type 2 diabetes. Retrospective and prospective observation studies were included. A total of nine studies met the inclusion criteria. Capillary β-OHB was found to have high sensitivity, specificity, positive predictive value and negative predictive value in identifying DKA compared to urinary ketone testing.

  5. Synthesis of Polynuclear Heterocyclic Compounds Derived from Thieno[2,3-d]pyrimidine Derivatives

    N. A. Hassan


    Full Text Available Reaction of 2-hydrazino-3-methyl-3,4-dihydrothieno[2,3-d]pyrimidin-4-one derivatives 2a,b with aliphatic acids afforded the thienotriazolopyrimidinone derivatives 3a-d, with nitrous acid yielded tetrazolothienopyrimidinone derivatives 4a,b and with carbon disulphide furnished 3-mercaptothienotriazolopyrimidinone derivatives 5a,b. Also, 2a,b reacted with aldehydes to afford the arylhydrazones 6a-f which cyclized into thienotriazolopyrimidinone derivatives 7a-f. Furthermore, 2a,b condensed with ethyl acetoacetate and ethyl cyanoacetate to afford 2-(1-pyrazolyl derivatives 9a,b and 10a,b, respectively. On the other hand, 2-hydrazino derivatives 2a,b condensed with a-halo-ketones to yield thienpyrimidotriazinone derivatives 11a,b and with β-diketones, to form 2-(1-pyrazolyl derivatives 12a-f.

  6. Effects of ketone bodies in Alzheimer's disease in relation to neural hypometabolism, β-amyloid toxicity, and astrocyte function

    Hertz, Leif; Chen, Ye; Waagepetersen, Helle S


    for the impaired glucose metabolism may be early destruction of the noradrenergic brain stem nucleus, locus coeruleus, which stimulates glucose metabolism, at least in astrocytes. These glial cells are essential in Alzheimer pathogenesis. The β-amyloid peptide Aβ interferes with their cholinergic innervation......Diet supplementation with ketone bodies (acetoacetate and β-hydroxybuturate) or medium-length fatty acids generating ketone bodies has consistently been found to cause modest improvement of mental function in Alzheimer's patients. It was suggested that the therapeutic effect might be more...... pronounced if treatment was begun at a pre-clinical stage of the disease instead of well after its manifestation. The pre-clinical stage is characterized by decade-long glucose hypometabolism in brain, but ketone body metabolism is intact even initially after disease manifestation. One reason...

  7. Dietary conjugated linoleic acid reciprocally modifies ketogenesis and lipid secretion by the rat liver.

    Sakono, M; Miyanaga, F; Kawahara, S; Yamauchi, K; Fukuda, N; Watanabe, K; Iwata, T; Sugano, M


    The effects of dietary conjugated linoleic acid (CLA) and linoleic acid (LA) on ketone body production and lipid secretion were compared in isolated perfused rat liver. After feeding the 1% CLA diet for 2 wk, the concentration of post-perfused liver cholesterol was significantly reduced by CLA feeding, whereas that of triacylglycerol remained unchanged. Livers from CLA-fed rats produced significantly more ketone bodies; and the ratio of beta-hydroxybutyrate to acetoacetate, an index of mitochondrial redox potential, tended to be consistently higher in the liver perfusate. Conversely, cumulative secretions of triacylglycerol and cholesterol were consistently lower in the livers of rats fed CLA, and the reduction in the latter was statistically significant. Thus dietary CLA appeared to exert its hypolipidemic effect at least in part through an enhanced beta-oxidation of fatty acids at the expense of esterification of fatty acid in the liver.

  8. Effects of insulin on ketogenesis following fasting in lean and obese men.

    Soeters, Maarten R; Sauerwein, Hans P; Faas, Linda; Smeenge, Martijn; Duran, Marinus; Wanders, Ronald J; Ruiter, An F; Ackermans, Mariëtte T; Fliers, Eric; Houten, Sander M; Serlie, Mireille J


    The ketone bodies (KBs) D-3-hydroxybutyrate (D-3HB) and acetoacetate (AcAc) play a role in starvation and have been associated with insulin resistance. The dose-response relationship between insulin and KBs was demonstrated to be shifted to the right in type 2 diabetes patients. However, KB levels have also been reported to be decreased in obesity. We investigated the metabolic adaptation to fasting with respect to glucose and KB metabolism in lean and obese men without type 2 diabetes using stable glucose and D-3HB isotopes in a two-step pancreatic clamp after 38 h of fasting. We found that D-3HB fluxes in the basal state were higher in lean compared to obese men: 15.2 (10.7-27.1) vs. 7.0 (3.5-15.1) micromol/kg lean body mass (LBM) x min, respectively, P ketogenesis. This implies differential insulin sensitivity of intermediary metabolism in obesity.

  9. Synthesis and Crystal Structure of Ethyl 2,7,7-Trimethyl-4-(4'-methylphenyl)-5-oxo-1,4,5,6,7,8-hexahydroquinoline-3-carboxylate in Aqueous Medium

    SHI Da-Qing; MOU Jie; ZHUANG Qi-Ya; WANG Xiang-Shan


    The title compound (C22H27NO3) was obtained by the reaction of 4-methylbenzal- dehyde, dimedone, ethyl acetoacetate and ammonium acetate in water in the presence of triethyl- benzylammonium chloride (TEBA). The crystal is of orthorhombic, space group Pbcn with a = 18.242(2), b = 15.651(1), c = 14.207(2) (A), V = 4056.15(64) (A)3, Mr = 353.45, Z = 8, Dc = 1.158 g/cm3, μ = 0.076, λ(MoKα) = 0.71073 (A) and F(000) = 1520. The final R = 0.0453 and wR = 0.0436 for 1156 observed reflections with I > 2((Ⅰ). X-ray analysis revealed that the pyridine ring is of boat conformation and the six-member ring fused with it adopts half-chair conformation.

  10. Identification by HPLC-MS of Anthocyanin Derivatives in Raisins

    Ana Marquez


    Full Text Available The anthocyanin composition in red grapes dried under controlled conditions has been studied. Pyranoanthocyanins and condensed anthocyanins with flavanols by a methylmethine bridge have been identified. Typically, these compounds appear in wine after the fermentation process, as they require compounds such as pyruvic acid, acetoacetic acid, and acetaldehyde for their formation. During the chamber-drying process a stress situation is originated, inducing significant changes in the grape metabolism from aerobic to anaerobic, and as a result it produces the activation of the alcohol dehydrogenase enzyme (ADH and others that would cause the formation of these compounds. These derivatives are very interesting because they give greater stability to the color of red wine.

  11. 双嘧达莫的合成工艺改进%Process Improvement on the Synthesis of Dipyridamole

    张秋荣; 蒋腾飞; 王惠; 邵坤鹏; 刘宏民


    Dipyridamole in total yield of 25.2% was synthesized by the condensation reaction of ethyl acetoacetate with thiourea, then nitration and oxidation, reduction, cyclization, chlorination, nucleo-philic substitution. The structure was confirmed by !H NMR and HR-MS.%以乙酰乙酸乙酯为原料,与硫脲缩合成嘧啶环后,经硝化和氧化、硝基还原、与尿素缩合生成2,4,6,8-四羟基嘧啶[5,4-d]并嘧啶,再经氯代、亲核取代等反应合成了抗血栓药双嘧达莫,总收率25.2%,其结构经1H NMR和HR-MS确证.

  12. Effect of carnitine on lipid metabolism in the newborn. I. Carnitine supplementation during total parenteral nutrition in the first 48 hours of life.

    Orzali, A; Donzelli, F; Enzi, G; Rubaltelli, F F


    The effect of carnitine administration on neonatal lipid metabolism was studied during endovenous loading with Intralipid (1 g/kg body weight over a 4-hour period). During a 6-hour period the plasma level of triglycerides, glycerol, free fatty acids (FFA), beta-hydroxybutyrate (beta-OHB), and acetoacetate were monitored in a group of newborns infused with carnitine and compared with a control group infused only with Intralipid. Carnitine administration caused an increased plasma concentration of ketone bodies, probably consequent to an increased rate of FFA mitochondrial beta-oxidation. An increased plasma level of glycerol and FFA was also observed, whereas the triglyceride plasma levels were not different between the two groups. Carnitine administration in the neonatal period seems to act by increasing ketogenesis and lipolysis.


    TANG Liming; GUO Wei; ZHOU Qixiang


    A polyacrylate/polyurethane (P(A)/P(U)) composite coating has been prepared by crosslinking an acetoacetylated polyacrylate with a vinylic group terminated polyurethane at room temperature. A model Michael reaction between ethyl acetoacetate (EAA) and methyl acrylate (MA) was designed to study the crosslinking mechanism. It was found that the two active hydrogen atoms in acetoacetyl group can both add to vinylic groups and the yield of mono- and bis-adducts are much affected by the molar ratio of acetoacetyl to vinylic groups. Higher crosslinking degree and better properties could be obtained with decreasing the molar ratio of the two active groups from 1/1 to 0.6/1 in the composite coatings.

  14. Ketone body β-hydroxybutyrate blocks the NLRP3 inflammasome-mediated inflammatory disease

    Youm, Yun-Hee; Nguyen, Kim Y.; Grant, Ryan W.; Goldberg, Emily L.; Bodogai, Monica; Kim, Dongin; D'Agostino, Dominic; Planavsky, Noah; Lupfer, Christopher; Kanneganti, Thirumala D.; Kang, Seokwon; Horvath, Tamas L.; Fahmy, Tarek M.; Crawford, Peter A.; Biragyn, Arya; Alnemri, Emad; Dixit, Vishwa Deep


    Ketone bodies , β-hydroxybutyrate (BHB) and acetoacetate support mammalian survival during states of energy deficit by serving as alternative source of ATP1. BHB levels are elevated during starvation, high-intensity exercise or by the low carbohydrate ketogenic diet2. Prolonged caloric restriction or fasting reduces inflammation as immune system adapts to low glucose supply and energy metabolism switches towards mitochondrial fatty acid oxidation, ketogenesis and ketolysis2-6. However, role of ketones bodies in regulation of innate immune response is unknown. We report that BHB, but neither acetoacetate nor structurally-related short chain fatty acids, butyrate and acetate, suppresses activation of the NLRP3 inflammasome in response to several structurally unrelated NLRP3 activators, without impacting NLRC4, AIM2 or non-canonical caspase-11 inflammasome activation. Mechanistically, BHB inhibits NLRP3 inflammasome by preventing K+ efflux and reducing ASC oligomerization and speck formation. The inhibitory effects of BHB on NLRP3 were not dependent on chirality or classical starvation regulated mechanisms like AMPK, reactive oxygen species (ROS), autophagy or glycolytic inhibition. BHB blocked NLRP3 inflammasome without undergoing oxidation in TCA cycle, independently of uncoupling protein-2 (UCP2), Sirt2, receptor Gpr109a and inhibition of NLRP3 did not correlate with magnitude of histone acetylation in macrophages. BHB reduced the NLRP3 inflammasome mediated IL-1β and IL-18 production in human monocytes. In vivo, BHB attenuates caspase-1 activation and IL-1β secretion in mouse models of NLRP3-mediated diseases like Muckle-Wells Syndrome (MWS), Familial Cold Autoinflammatory syndrome (FCAS) and urate crystal induce body cavity inflammation. Taken together, these findings suggest that the anti-inflammatory effects of caloric restriction or ketogenic diets may be mechanistically linked to BHB-mediated inhibition of the NLRP3 inflammasome, and point to the potential

  15. Preparation of TiO2-C Nano-composite Materials by Sol-Gel Method%溶胶-凝胶法制备TiO2-C纳米复合材料的研究

    邵霞; 张睿; 陆文聪; 卢春


    TiO2-C nanocomposite materials were prepared by using sol-gel process, which were characterized by XRD, SEM and TEM. Results showed that the precursors of TiO2 and carbon could be co-gelated soon at room temperature by adding propylene oxide as gelation initiator. TiO2 grain grows up after it is heated at 600 ℃ for 3 h, and its characteristic peak is more clear. The crystal structure of TiO2 is anatase. Ethyl acetoacetate as a complexing agent is beneficial for the formation of the gels. The pore volume and the pore diameter in the TiO2-C nanocomposite materials increase with the increasing amount of the ethyl acetoacetate.%采用溶胶凝胶法制备了TiO2-C纳米复合材料,利用XRD、SEM和TEM等方法对样品进行分析表征.实验结果表明:网络诱导剂的加入有利于样品在室温下静置后更快凝胶,经600℃焙烧3h后样品中二氧化钛晶粒有不同 程度长大,其特征峰更加明显,且其中二氧化钛晶粒为锐钛矿型;乙酰乙酸乙酯的加入有利于溶胶-凝胶的形成,且随着乙酰乙酸乙酯加入量的增加,TiO2-C纳米复合材料的孔容和孔径均增加.

  16. Stereochemical features of unsaturated aminodiesters - intermediates in the synthesis of Richlocaine

    Meyrambek Ospanov


    Full Text Available The article provides information on the methods for synthesis of unsaturated aminodiesters, which are the key intermediates in the synthesis of N-substituted 2,5-dimethyl-piperidin-4-ones, including 1,2,5-trimetilpieridin-3-one. The developed method includes reaction of methyl ester of methacrylic acid with alkyl (alkenyl amines. The obtained N-alkyl(alkenyl-(2-methyl-2- carbmetoxyethyl methylamine are attached to acetoacetic ester being in the enol form. The optimal conditions for synthesis of N-alkyl(alkenyl-(2-methyl-2-carbethoxyethenemethylamines have been established. Results of gas chromatographic analysis has showed that, at room temperature, the reaction in polar solvents is delayed for two days. In non-polar solvents, the reaction runs with simultaneous distillation of water as an azeotrope with the solvent. Increase of the radical size at N atom has little effect on the reaction rate. Introduction of allyl radical at the nitrogen atom, apparently due to steric hindrance, resulted in lower reactivity of the monoester with allyl radical by condensation with acetoacetic ester. The structure of (1-methyl-2-carbethoxyethene-(2-methyl-2-carbmethoxyethyl methylamine has been studied by PMR spectroscopy. Study of the effect of chirality on the NMR spectra showed that, if the molecule of the organic compound has an asymmetric carbon atom, it may lead to the disappearance of the magnetic equivalence of neighboring protons or group of protons. Presence of an asymmetric carbon atom in b-position to the amino group of enamine leads to the disappearance of the magnetic equivalence of the neighboring protons is observed in the form of two quartets. For enaminodiesters, equilibrium is shifted toward the trans-S-cis conformer where less steric hindrance is pronounced.

  17. Preparation of functionalized graphene oxide and its application as a nanoadsorbent for Hg(2+) removal from aqueous solution.

    Aghdam, Khaledeh; Panahi, Homayon Ahmad; Alaei, Ebrahim; Hasani, Amir Hesam; Moniri, Elham


    A poly(allyl acetoacetate)-grafted graphene oxide (GO-GAA) was successfully synthesized using Hummer's method by divinyl sulfone modification and allyl acetoacetate polymerizaton. This novel functionalized graphene oxide was characterized thoroughly by FTIR, XRD, FE-SEM, TEM, and TG-DT analyses. GO-GAA was then employed as an adsorbent for Hg(2+) removal from aqueous solutions. It exhibited higher adsorption capacity with regard to the pristine graphene oxide because of its effective functionalities, especially the dicarbonyl groups which are significant chelating agents. The effects of pH, temperature, and contact time on Hg(2+) adsorption were also investigated. The optimum Hg(2+) adsorption was obtained at pH 4 and T = 20-30 °C. The adsorption isotherm and kinetics were found to follow the Langmuir and pseudo-second-order models, respectively, with a correlation coefficient of 0.99 for both. The calculated maximum adsorption capacity of the adsorbent was 282.7 mg Hg(2+) per unit mass of GO-GAA, which is much more than 56 mg/g of that obtained for GO. The results showed that adsorption reaches up to 95 % of its maximum in less than 2 min. The synthesized GO-GAA as a novel and efficient adsorbent has been regenerated by HNO3 and reused. It retained its performance for Hg(2+) removal for several times and a less than 5 % decrease in removal efficiency was observed after four cycles of adsorption-desorption.

  18. Inhibition of the signal transduction through the AtoSC system by histidine kinase inhibitors in Escherichia coli.

    Theodorou, Evaggelos C; Theodorou, Marina C; Kyriakidis, Dimitrios A


    AtoSC two-component system participates in many indispensable processes of Escherichia coli. We report here that the AtoSC signal transduction is inhibited by established histidine kinase inhibitors. Closantel, RWJ-49815 and TNP-ATP belonging to different chemical classes of inhibitors, abrogated the in vitro AtoS kinase autophosphorylation. However, when AtoS was embedded in the membrane fractions, higher inhibitor concentrations were required for total inhibition. When AtoS interacted with AtoC forming complex, the intrinsic histidine kinase was protected by the response regulator, requiring increased inhibitors concentrations for partially AtoS autophosphorylation reduction. The inhibitors exerted an additional function on AtoSC, blocking the phosphotransfer from AtoS to AtoC, without however, affecting AtoC~P dephosphorylation. Their in vivo consequences through the AtoSC inhibition were elucidated on atoDAEB operon expression, which was inhibited only in AtoSC-expressing bacteria where AtoSC was induced by acetoacetate or spermidine. The inhibitor effects were extended on the AtoSC regulatory role on cPHB [complexed poly-(R)-3-hydroxybutyrate] biosynthesis. cPHB was decreased upon the blockers only in acetoacetate-induced AtoSC-expressing cells. Increased ATP amounts during bacterial growth reversed the inhibitory TNP-ATP-mediated effect on cPHB. The alteration of pivotal E. coli processes as an outcome of AtoSC inhibition, establish this system as a target of two-component systems inhibitors.

  19. The role of insulin, glucagon, dexamethasone, and leptin in the regulation of ketogenesis and glycogen storage in primary cultures of porcine hepatocytes prepared from 60 kg pigs.

    Fernández-Fígares, I; Shannon, A E; Wray-Cahen, D; Caperna, T J


    A study was conducted to elucidate hormonal control of ketogenesis and glycogen deposition in primary cultures of porcine hepatocytes. Hepatocytes were isolated from pigs (54-68 kg) by collagenase perfusion and seeded into collagen-coated T-25 flasks. Monolayers were established in medium containing fetal bovine serum for 1 day and switched to a serum-free medium for the remainder of the culture period. Hepatocytes were maintained in DMEM/M199 containing 1% DMSO, dexamethasone (10(-6) or 10(-7) M), linoleic acid (3.4 x 10(-5) M), and carnitine (10(-3) M) for 3 days. On the first day of serum-free culture, insulin was added at 1 or 100 ng/ml and glucagon was added at 0, 1, or 100 ng/ml. Recombinant human leptin (200 ng/ml) was added during the final 24 h; medium and all cells were harvested on the third day. Concentrations of acetoacetate and beta-hydroxybutyrate (ketone bodies) in media and glycogen deposition in the cellular compartment were determined. Ketogenesis was highly stimulated by glucagon (1 and 100 ng/ml) and inhibited by insulin. In contrast, glycogen deposition was stimulated by insulin and attenuated by glucagon; high insulin was also associated with a reduction in the ketone body ratio (acetoacetate:beta-hydroxybutyrate). High levels of dexamethasone stimulated ketogenesis, but inhibited glycogen deposition at low insulin. Culture of cells with leptin for 24 h, over the range of insulin, glucagon, and dexamethasone concentrations had no effect on either glycogen deposition or ketogenesis. These data suggest that while adult porcine hepatocytes are indeed sensitive to hormonal manipulation, leptin has no direct influence on hepatic energy metabolism in swine.

  20. Protein and amino acid metabolism in skeletal muscle

    Wu, Guoyao.


    Isolated chick extensor digitorum communis (EDC) muscles and, in some experiments, rat skeletal muscles were used to study a number of aspects of protein and amino acid metabolism. (1) Chick EDC muscles synthesize and release large amounts of alanine and glutamine, which indirectly obtain their amino groups from branched-chain amino acids (BCAA). (2) Acetoacetate or DL-{beta}-hydroxybutyrate (4 mM) decrease (P < 0.01) alanine synthesis and BCAA transamination in EDC muscles from 24-h fasted chicks by decreasing (P < 0.01) intracellular concentrations of pyruvate due to inhibition of glycolysis. (3) Glutamine is extensively degraded in skeletal muscles from both chicks and rats, thus challenging the traditional view that glutamine oxidation is negligible in skeletal muscle. The cytosolic glutamine aminotransferases L and K in the rat and the mitochondrial phosphate-activated glutaminase in the chick play important roles in the conversion of glutamine to {alpha}-ketoglutarate for further oxidation. (4) Although methionine has been reported to be extensively transaminated in rat skeletal muscle preparations in the absence of other amino acids, transamination of methionine is absent or negligible in chick and rat skeletal muscles in the presence of physiological concentrations of amino acids. (5) Glutamine at 1.0-15 mM increases (P < 0.01) protein synthesis ({sup 3}H-phenylalanine incorporation), and at 10.0-15.0 mM decreases (P < 0.05) protein degradation ({sup 3}H-phenylalanine release from prelabelled protein in vivo) in EDC muscles from fed chicks as compared to muscles incubated in the absence of glutamine. (6) Acetoacetate or DL-{beta}-hydroxybutyrate (4 mM) has a small but significant inhibitory effect (P < 0.05) on the rate of protein synthesis, but has no effect (P > 0.05) on the rate of protein degradation in EDC muscles from fed chicks.

  1. Amiodarone inhibits the mitochondrial beta-oxidation of fatty acids and produces microvesicular steatosis of the liver in mice

    Fromenty, B.; Fisch, C.; Labbe, G.; Degott, C.; Deschamps, D.; Berson, A.; Letteron, P.; Pessayre, D. (Institut National de la Sante et de la Recherche Medicale U24, Clichy (France))


    Amiodarone has been shown to produce microvesicular steatosis of the liver in some recipients. We have determined the effects of amiodarone on the mitochondrial oxidation of fatty acids in mice. In vitro, the formation of 14C-acid-soluble beta-oxidation products from (U-14C)palmitic acid by mouse liver mitochondria was decreased by 92% in the presence of 125 microM amiodarone and by 94% in the presence of 125 microM N-desethylamiodarone. Inhibition due to 100 or 150 microM amiodarone persisted in the presence of 5 mM acetoacetate, whereas acetoacetate totally relieved inhibition due to 15 microM rotenone. In vivo, exhalation of (14C)CO2 from (U-14C)palmitic acid was decreased by 31, 40, 58 and 78%, respectively, in mice receiving 19, 25, 50 and 100 of amiodarone hydrochloride 1 hr before the administration of (U-14C)palmitic acid. One hour after 100, the exhalation of (14C)CO2 from (1-14C)palmitic acid, (1-14C)octanoic acid or (1-14C)butyric acid was decreased by 78, 72 and 53%, respectively. Exhalation of (14C)CO2 from (1-14C)palmitic acid was normal between 6 and 9 hr after administration of 100 of amiodarone hydrochloride, but was still inhibited by 71 and 37%, 24 and 48 hr after 600 Twenty four hours after the latter dose of amiodarone, hepatic triglycerides were increased by 150%, and there was microvesicular steatosis of the liver. We conclude that amiodarone inhibits the mitochondrial beta-oxidation of fatty acids and produces microvesicular steatosis of the liver in mice.

  2. Enhanced lipid utilization in infants receiving oral L-carnitine during long-term parenteral nutrition.

    Helms, R A; Whitington, P F; Mauer, E C; Catarau, E M; Christensen, M L; Borum, P R


    Fourteen infants requiring long-term total parenteral nutrition but able to tolerate small quantities of enteral feedings were randomized into carnitine treatment and placebo control groups. All infants had received nutritional support devoid of carnitine. Plasma carnitine levels and observed plasma lipid indices were not different before supplementation. Under standardized, steady-state conditions, 0.5 g/kg fat emulsion (intralipid) was administered intravenously over 2 hours both before and after infants received 7 days of continuous nasogastric or gastric tube L-carnitine (50 mumol/kg/day) or placebo. Plasma triglyceride, free fatty acid, acetoacetate, beta-hydroxybutyrate, and carnitine concentrations were observed at 0 (start of lipid infusion), 2, and 4 hours for pre- and post-treatment periods, and in addition at 6 and 8 hours after carnitine supplementation. Infants receiving carnitine had significantly greater beta-hydroxybutyrate plasma concentrations (P less than 0.05) and carnitine (P less than 0.001) at 0, 2, 4, 6, and 8 hours, and greater plasma acetoacetate concentrations (P less than 0.05) at 2, 4, 6, and 8 hours, compared with controls. Twenty-four-hour urinary carnitine excretion was very low for both groups before supplementation; after supplementation, excretion was higher (P less than 0.05) in the carnitine group. No significant differences were found between groups for plasma triglyceride or free fatty acid concentrations at any observation period. This study demonstrated enhanced fatty acid oxidation, as evidenced by increased ketogenesis, with L-carnitine supplementation in infants receiving long-term total parenteral nutrition.

  3. Blister Test for Measurements of Adhesion and Adhesion Degradation of Organic Polymers on AA2024-T3

    Rincon Troconis, Brendy Carolina

    cleaning/desmutting and conversion coating (CC) on the adhesion strength of acetoacetate to AA2024-T3 and the effects of improper water rinse temperature after cleaner were assessed using the BT. The results showed that pretreatments improve the adhesion strength of acetoacetate primer on AA2024-T3, but the comparative behavior depends on the specific treatment. Process control is of paramount importance for the performance of acetoacetate coated systems applied on AA2024-T3. The lack of thermal activity in the water rinse after cleaning step produces deleterious effects on the adhesion and blistering resistance of CC. Finally, a test sample incorporating a coated and scribed Al alloy panel and uncoated through-hole fasteners (Ti, SS316, AA2024-T3) was designed to provide accelerated response during atmospheric corrosion testing in the field (long-term beach exposure) or in laboratory chambers (ASTM B117). The results after only three weeks of exposure to ASTM B117 correlated well with long-term beach exposure, allowing rapid ranking of different coating systems. Of the fastener materials studied, visual observation and volume lost determination indicated that the worst attack occurred near SS316 fasteners its effect was explained by Scanning Kelvin Probe measurements and the available cathodic current measured in chloride solution.

  4. Study on sedative, hypnotic and effects of pine needle on neurotransmitter%松针不同提取物镇静催眠作用及对脑神经递质的影响

    赵桂芝; 寿旦; 俞忠明; 李洪玉


    目的:研究比较松针不同提取物的镇静、催眠及对脑神经递质的作用.方法:通过直接诱导小鼠睡眠,观察比较松针不同提取物对戊巴比妥钠阈上、阈下剂量致小鼠睡眠的协同作用;采用HPLC法测定给药后小鼠脑海马中γ-氨基丁酸和谷氨酸的含量.结果:松针正丁醇提取物和乙酸乙酯提取物能明显延长阈上剂量戊巴比妥钠致小鼠睡眠时间(P<0.01),增加阈下剂量戊巴比妥钠致小鼠睡眠只数(P<0.01);有效提取物可增加海马中γ-氨基丁酸含量(P<0.05).结论:松针正丁醇提取物和乙酸乙酯提取物均具有明显的镇静催眠作用.%Objective: To study the sedative and hypnotic effects of the different extracts of pine needle and the effect on neurotransmitter. Methods: Sedative and hypnotic experiments were made to observe the effects of different extracts of pine needle on spontaneous motion and hypnosis in mice. Determining the levels of aminobutyric acid and glutamic acid in administered mice of hippocamp by HPLC. Results: The n-butanol extract of pine needle and its acetoacetate extract could enhance the sleeping number of pentobarbital sodium (subthleshold dosage) and prolong the sleeping time of sodium pentobarbital (superthleshold dosage, P<0.01); Effective extract can affect neurotransmitter in hippocampal slices (P<0.05). Conclusion: The n-butanol extract of pine needle and its acetoacetate extract had sedative and hypnotic effect.

  5. On Michael Addition Reactions of Chalcones%查尔酮类化合物的 Michael加成反应

    张鹏会; 李艳春; 蒲雯


    A series of chalcones are synthesized from aromatic aldehyde and acetophenone ,with H2 O as the solvent and sodium hydroxide as the catalyst .A series of Michael addition products are obtained from these chalcones and ethyl acetoacetate , catalyzed by sodium hydroxide and anhydrous sodium carbonate under the assistance of ultrasonic irradiation .And the inflences of the addition reaction condition on the yield are studied .The premium condition are determined by followings:the molar ratio of chalcone and ethyl acetoacetate is 1/1.5,sodium hydroxide/anhydrous sodium carbonate is 1/1.5 under PEG -400/H2O in 40oC,the desired product is obtained,and the yield can be up to 91.2%.The method of this syn-thesis has advantages such as simple to operate ,short reaction time and high yield .%以芳香醛与苯乙酮为原料, NaOH 为催化剂,在水相中合成了一系列查尔酮化合物。在超声辅助下,研究了混合碱( NaOH-Na2 CO3)催化查尔酮与三乙(乙酰乙酸乙酯)的 Michael加成反应,合成了一系列Michael加成产物,并对影响反应的因素进行了探究,得出最佳反应条件为:40oC 下,在 PEG -400/H2O 介质中,n查尔酮/n三乙=1/1.5,nNaOH/nNa2CO3=1/1.5,得到目标产物的产率最好,高达91.2%。此方法操作简单,反应时间短,产率高。

  6. [Methylglyoxal--a test for impaired biological functions of exotrophy and endoecology, low glucose level in the cytosol and gluconeogenesis from fatty acids (a lecture)].

    Titov, V N; Dmitriev, L F; Krylin, V A; Dmitriev, V A


    In philogenesis, due to the failure to store a great deal of carbohydrates in vivo as glycogen, all animal species began synthesizing from glucose palminitic fatty acid and depositing it as triglycerides. During biological dysfunction of exotrophy (long starvation, early postnatality, hibernation), cells also accomplish a reverse synthesis of glucose from fatty acids under aerobic conditions. Under physiological conditions, acetyl-CoA that is converted to malate and pyruvate in the glyoxalate cycle is a substrate of glyconeogenesis. Under pathological conditions of hypoxia and deficiency of macroerges, gluconeogenesis occurs without ATP consumption through the methylglyoxal pathway when used as a substrate of ketone bodies via the pathway: butyric acid (butyrate) --> beta-hydroxybutyrate --> acetoacetate --> acetone --> acetol --> methylglyoxal --> S-D-lactol-glutathione --> D-lactate --> pyruvate --> D-lactate. Under physiological conditions, this gluconeogenesis pathway does not function. We believe that with low glucose levels in the cell cytosole (glycopenia), under pathological conditions of hypoxia and due to failure to mitochondria to oxidize fatty acids, gene expression and gluconeogenesis occur through the methylglyoxal pathway. At the same time, the cytosol, intercellular environment, and plasma shows the elevated levels of methylglyoxal and D-lactate that it is converted to by the action of glyoxalases I and II. Under pathological conditions, glycopenia develops in starvation, diabetes, and metabolic acidosis, neoplasms, renal failure, and possibly, metabolic syndrome. The chemical interaction of methylglyoxal with the amino acid residues of lysine and arginine results in the denaturation of circulating and structurized proteins via carbonylation--glycosylation.

  7. Biginelli Condensation Assisted by Microwave Irradiation in Ionic Liquids%离子液体中微波促进的Biginelli缩合反应



    The reaction of a one-pot three component Biginelli condensation can be dramatically accelerated by irradiating the reaction mixture of ethyl acetoacetate, aryl aldehyde and urea in an ionic liquid 1-butyl-3-methylimidazonium tetrafluoroborate([bmim]BF4) as a catalyst with a Microwave oven. Ten 5-ethoxycarbonyl-4-aryldihydropyrimidinones were synthesized in 10min with excellent yields. Their structures were comfirmed by IR and 1H NMR.%在微波辐射下,以离子液体1-丁基-3-甲基咪唑四氟硼酸盐([bmim]BF4)为催化剂,三组分一锅法Bigi-nelli缩合反应快速地合成了10种高收率的5-乙氧羰基-4-芳基二氢嘧啶酮衍生物的缩合产物,其结构经IR和1H NMR确证.

  8. Sol-gel-derived hybrid materials multi-doped with rare-earth metal ions

    Zelazowska, E.; Rysiakiewicz-Pasek, E.; Borczuch-Laczka, M.; Cholewa-Kowalska, K.


    Four different hybrid organic-inorganic materials based on TiO2-SiO2 matrices with organic additives and doped with rare-earth metal ions (III) from the group of europium, cerium, terbium, neodymium, dysprosium and samarium, were synthesized by sol-gel method. Tetraethyl orthosilicate, titanium (IV) isopropoxide and organic compounds, such as butyl acrylate, butyl methacrylate, ethyl acetoacetate, ethylene glycol dimethacrylate, ethyl acetate, propylene carbonate, organic solvents and certain inorganic salts were used in the synthesis. The inorganic part of the sols, which were used in the synthesis of all the hybrid materials, was prepared separately and then the organic parts were added. The materials obtained were aged for three weeks at room temperature and then heated in an electric oven for three hours at temperatures of 80 °C-150 °C. Scanning electron microscopy equipped with energy dispersive X-ray spectroscopy (SEM/EDX); X-ray diffraction (XRD); Fourier transform infrared spectroscopy (KBr technique); 29Si magic-angle spinning nuclear magnetic resonance; and fluorescence spectroscopy were used for the examination of morphology, microstructure and luminescence properties, respectively. Photoluminescence properties with relatively intense narrow emission lines of Tb, Eu, Dy, Nd, Sm respectively to the RE-ions doping, were observed for all the hybrid materials.

  9. cBSA-147 for the preparation of bacterial biofilms in a microchannel reactor.

    Ng, Jeck Fei; Jaenicke, Stephan; Eisele, Klaus; Dorn, Jan; Weil, Tanja


    Whole cells are attractive biocatalysts, particularly if the reaction requires cofactors or involves multiple transformations. Immobilization of the catalyst is often a prerequisite for continuous processes. The highly cationic chemically modified plasma protein bovine serum albumin (cBSA-147) has been applied for the electrostatically mediated immobilization of the planktonic bacterium E. coli BL21 star (DE3), and the resulting biofilms were superior to those formed on poly-L-lysine coated surfaces. The biocatalyst was immobilized in a capillary column (inside diameter of 530 μm and L=30 m) and evaluated in the enantioselective reduction of ethyl acetoacetate to R-(-)ethyl hydroxybutyrate. In continuous operation in the microreactor format, the productivity of the cells was about 30% higher than that determined in a bench-scale fermentation system. This increase is attributed to the improved mass transfer over short geometrical dimensions. The similarity in the results indicates that studies on a biofilm-coated microreactor can be used for the accelerated collection of data for process optimization.

  10. Urinary Metabolomic Approach Provides New Insights into Distinct Metabolic Profiles of Glutamine and N-Carbamylglutamate Supplementation in Rats.

    Liu, Guangmang; Cao, Wei; Fang, Tingting; Jia, Gang; Zhao, Hua; Chen, Xiaoling; Wu, Caimei; Wang, Jing


    Glutamine and N-carbamylglutamate can enhance growth performance and health in animals, but the underlying mechanisms are not yet elucidated. This study aimed to investigate the effect of glutamine and N-carbamylglutamate supplementation in rat metabolism. Thirty rats were fed a control, glutamine, or N-carbamylglutamate diet for four weeks. Urine samples were analyzed by nuclear magnetic resonance (NMR)-based metabolomics, specifically high-resolution ¹H NMR metabolic profiling combined with multivariate data analysis. Glutamine significantly increased the urine levels of acetamide, acetate, citrulline, creatinine, and methymalonate, and decreased the urine levels of ethanol and formate (p carbamylglutamate significantly increased the urine levels of creatinine, ethanol, indoxyl sulfate, lactate, methymalonate, acetoacetate, m-hydroxyphenylacetate, and sarcosine, and decreased the urine levels of acetamide, acetate, citrulline, creatine, glycine, hippurate, homogentisate, N-acetylglutamate, phenylacetyglycine, acetone, and p-hydroxyphenylacetate (p carbamylglutamate could modify urinary metabolome related to nitrogen metabolism and gut microbiota metabolism. Moreover, N-carbamylglutamate could alter energy and lipid metabolism. These findings indicate that different arginine precursors may lead to differences in the biofluid profile in rats.

  11. Synthesis of 2(5H)-pyrrinones and the inhibitory effects on the Biofilms of P.aeruginosa in vitro%2(5H)-吡咯酮衍生物的合成及对铜绿假单胞菌生物膜的体外作用

    郭嘉亮; 刘兆祥; 巢伟; 孙平华


    以乙酰乙酸乙酯为原料,设计合成2(5H)-吡咯酮及其衍生物,所得化合物经1H NMR、13C NMR、MS和元素分析等确证.通过扫描电镜观察2(5H)-吡咯酮对黏液型铜绿假单胞菌P.aeruginosa(PA)体外生物被膜(biofilms,BF)的影响,可见BF被破坏,基质样物变稀疏,细菌群聚大为降低.活性实验结果表明,2(5H)-吡咯酮类化合物具有一定的抑制细菌群体感应的能力.%A series of pyrroles and 2 (5H) -pyrrinones were successfully synthesized from ethyl aceto-acetate, and characterized by 'H NMR, 13C NMR, MS spectra and elemental analysis. Scanning electron microscope (SEM) was used to evaluate the effects of the 2 (5H) -pyrrolones on the quorum sensing (QS) system and the bacterial biofilm (BF) formation biofilm formation. The biofilms of P. aeruginosa were inhibited, the base became sparseness, and the swarming was decreased. The results of biological activity tests indicated that 2 (5H) -pyrrinones showed potent ability to inhibit the formation of bacterial biofilms.

  12. Anti-Diabetic Activity and Metabolic Changes Induced by Andrographis paniculata Plant Extract in Obese Diabetic Rats

    Muhammad Tayyab Akhtar


    Full Text Available Andrographis paniculata is an annual herb and widely cultivated in Southeast Asian countries for its medicinal use. In recent investigations, A. paniculata was found to be effective against Type 1 diabetes mellitus (Type 1 DM. Here, we used a non-genetic out-bred Sprague-Dawley rat model to test the antidiabetic activity of A. paniculata against Type 2 diabetes mellitus (Type 2 DM. Proton Nuclear Magnetic Resonance (1H-NMR spectroscopy in combination with multivariate data analyses was used to evaluate the A. paniculata and metformin induced metabolic effects on the obese and obese–diabetic (obdb rat models. Compared to the normal rats, high levels of creatinine, lactate, and allantoin were found in the urine of obese rats, whereas, obese-diabetic rats were marked by high glucose, choline and taurine levels, and low lactate, formate, creatinine, citrate, 2-oxoglutarate, succinate, dimethylamine, acetoacetate, acetate, allantoin and hippurate levels. Treatment of A. paniculata leaf water extract was found to be quite effective in restoring the disturbed metabolic profile of obdb rats back towards normal conditions. Thisstudy shows the anti-diabetic potential of A. paniculata plant extract and strengthens the idea of using this plant against the diabetes. Further classical genetic methods and state of the art molecular techniques could provide insights into the molecular mechanisms involved in the pathogenesis of diabetes mellitus and anti-diabetic effects of A. paniculata water extract.

  13. Polyunsaturated fatty acyl-coenzyme As are inhibitors of cholesterol biosynthesis in zebrafish and mice

    Santhosh Karanth


    Lipid disorders pose therapeutic challenges. Previously we discovered that mutation of the hepatocyte β-hydroxybutyrate transporter Slc16a6a in zebrafish causes hepatic steatosis during fasting, marked by increased hepatic triacylglycerol, but not cholesterol. This selective diversion of trapped ketogenic carbon atoms is surprising because acetate and acetoacetate can exit mitochondria and can be incorporated into both fatty acids and cholesterol in normal hepatocytes. To elucidate the mechanism of this selective diversion of carbon atoms to fatty acids, we fed wild-type and slc16a6a mutant animals high-protein ketogenic diets. We find that slc16a6a mutants have decreased activity of the rate-limiting enzyme of cholesterol biosynthesis, 3-hydroxy-3-methylglutaryl-coenzyme A reductase (Hmgcr, despite increased Hmgcr protein abundance and relative incorporation of mevalonate into cholesterol. These observations suggest the presence of an endogenous Hmgcr inhibitor. We took a candidate approach to identify such inhibitors. First, we found that mutant livers accumulate multiple polyunsaturated fatty acids (PUFAs and PUFA-CoAs, and we showed that human HMGCR is inhibited by PUFA-CoAs in vitro. Second, we injected mice with an ethyl ester of the PUFA eicosapentaenoic acid and observed an acute decrease in hepatic Hmgcr activity, without alteration in Hmgcr protein abundance. These results elucidate a mechanism for PUFA-mediated cholesterol lowering through direct inhibition of Hmgcr.

  14. Design, synthesis of novel pyranotriazolopyrimidines and evaluation of their anti-soybean lipoxygenase, anti-xanthine oxidase, and cytotoxic activities.

    Saïd, Abderrahim Ben; Romdhane, Anis; Elie, Nicolas; Touboul, David; Jannet, Hichem Ben; Bouajila, Jalloul


    The synthesis of 14-(aryl)-14H-naphto[2,1-b]pyrano[3,2-e][1,2,4]triazolo[1,5-c]pyrimidine-2-yl) acetamidoximes 2a-e has been accomplished by reaction of 2-acetonitrile derivatives 1a-e with hydroxylamine. Cyclocondensation reaction of precursors 2a-e with some elctrophilic species such as ethylorthoformate, acetic anhydride, and methyl-acetoacetate provided the new oxadiazole derivatives 3a-e, 4a-e, and 5a-e, respectively. On the other hand, the reaction of precursors 2a-e with 2-chloropropanoyl chloride afforded the new acetimidamides 6a-e which evolve under reflux of toluene to the new oxadiazoles 7a-e. The synthetic compounds were screened for their anti-xanthine oxidase, anti-soybean lipoxygenase, and cytotoxic activities. Moderate to weak xanthine oxidase and soybean lipoxygenase inhibitions were obtained but significant cytotoxic activities were noted. The most cytotoxic activities were recorded mainly (i) 5a was the most active (IC50 = 4.0 μM) and selective against MCF-7 and (ii) 2a was cytotoxic against the four cell lines with selectivity for MCF-7 and OVCAR-3 (IC50 = 17 and 12 μM, respectively) while 2e is highly selective against OVCAR-3 (IC50 = 10 μM).

  15. Metabolic engineering of Candida utilis for isopropanol production.

    Tamakawa, Hideyuki; Mita, Tokiko; Yokoyama, Aki; Ikushima, Shigehito; Yoshida, Satoshi


    A genetically-engineered strain of the yeast Candida utilis harboring genes encoding (1) an acetoacetyl-CoA transferase from Clostridium acetobutylicum ATCC 824, (2) an acetoacetate decarboxylase, and (3) a primary-secondary alcohol dehydrogenase derived from Clostridium beijerinckii NRRL B593 produced up to 0.21 g/L of isopropanol. Because the engineered strain accumulated acetate, isopropanol titer was improved to 1.2 g/L under neutralized fermentation conditions. Optimization of isopropanol production was attempted by the overexpression and disruption of several endogenous genes. Simultaneous overexpression of two genes encoding acetyl-CoA synthetase and acetyl-CoA acetyltransferase increased isopropanol titer to 9.5 g/L. Moreover, in fed-batch cultivation, the resultant recombinant strain produced 27.2 g/L of isopropanol from glucose with a yield of 41.5 % (mol/mol). This is the first demonstration of the production of isopropanol by genetically engineered yeast.

  16. Modifying the product pattern of Clostridium acetobutylicum: physiological effects of disrupting the acetate and acetone formation pathways.

    Lehmann, Dörte; Hönicke, Daniel; Ehrenreich, Armin; Schmidt, Michael; Weuster-Botz, Dirk; Bahl, Hubert; Lütke-Eversloh, Tina


    Clostridial acetone-butanol-ethanol (ABE) fermentation is a natural source for microbial n-butanol production and regained much interest in academia and industry in the past years. Due to the difficult genetic accessibility of Clostridium acetobutylicum and other solventogenic clostridia, successful metabolic engineering approaches are still rare. In this study, a set of five knock-out mutants with defects in the central fermentative metabolism were generated using the ClosTron technology, including the construction of targeted double knock-out mutants of C. acetobtuylicum ATCC 824. While disruption of the acetate biosynthetic pathway had no significant impact on the metabolite distribution, mutants with defects in the acetone pathway, including both acetoacetate decarboxylase (Adc)-negative and acetoacetyl-CoA:acyl-CoA transferase (CtfAB)-negative mutants, exhibited high amounts of acetate in the fermentation broth. Distinct butyrate increase and decrease patterns during the course of fermentations provided experimental evidence that butyrate, but not acetate, is re-assimilated via an Adc/CtfAB-independent pathway in C. acetobutylicum. Interestingly, combining the adc and ctfA mutations with a knock-out of the phosphotransacetylase (Pta)-encoding gene, acetate production was drastically reduced, resulting in an increased flux towards butyrate. Except for the Pta-negative single mutant, all mutants exhibited a significantly reduced solvent production.

  17. Butenolide inhibits marine fouling by altering the primary metabolism of three target organisms

    Zhang, Yifan


    Butenolide is a very promising antifouling compound that inhibits ship hull fouling by a variety of marine organisms, but its antifouling mechanism was previously unknown. Here we report the first study of butenolides molecular targets in three representative fouling organisms. In the barnacle Balanus (=Amphibalanus) amphitrite, butenolide bound to acetyl-CoA acetyltransferase 1 (ACAT1), which is involved in ketone body metabolism. Both the substrate and the product of ACAT1 increased larval settlement under butenolide treatment, suggesting its functional involvement. In the bryozoan Bugula neritina, butenolide bound to very long chain acyl-CoA dehydrogenase (ACADVL), actin, and glutathione S-transferases (GSTs). ACADVL is the first enzyme in the very long chain fatty acid β-oxidation pathway. The inhibition of this primary pathway for energy production in larvae by butenolide was supported by the finding that alternative energy sources (acetoacetate and pyruvate) increased larval attachment under butenolide treatment. In marine bacterium Vibrio sp. UST020129-010, butenolide bound to succinyl-CoA synthetase β subunit (SCSβ) and inhibited bacterial growth. ACAT1, ACADVL, and SCSβ are all involved in primary metabolism for energy production. These findings suggest that butenolide inhibits fouling by influencing the primary metabolism of target organisms. © 2012 American Chemical Society.

  18. Synthesis, anti-inflammatory, analgesic and anticonvulsant activities of some new 4,6-dimethoxy-5-(heterocyclesbenzofuran starting from naturally occurring visnagin

    E.R. El-Sawy


    Full Text Available Novel 3-(4,6-dimethoxybenzofuran-5-yl-1-phenyl-1H-pyrazole-4-carboxaldehyde (3 and 3-chloro-3-(4,6-dimethoxybenzofuran-5-ylpropenal (4 were prepared via Vilsmeier–Haack reaction of 1-(4,6-dimethoxybenzofuran-5-ylethanone (1 and its hydrazone derivative 2. Reaction of compound 4 with some hydrazine derivatives, namely hydrazine hydrate, phenylhydrazine and benzylhydrazine hydrochloride led to the formation of pyrazole derivatives 5–8, respectively. On the other hand, reaction of compound 4 with thiourea, urea or guanidine gave the pyrimidine derivatives 9–11, respectively. Reaction of amino compound 11 with acetic anhydride, benzoyl chloride and benzenesulphonyl chloride yielded N-substituted pyrimidine derivatives 12–14, respectively. Reaction of diazonium salt of compound 11 with sodium azide afforded azidopyrimidine derivative 15, which upon reaction with ethyl acetoacetate gave 1,2,3-triazole derivative 16. Acid catalyzed reaction of 11 with p-nitrobenzaldehyde gave Schiff base 17, which cyclized upon reaction with thioglycolic acid or chloroacetyl chloride to give thiazolidin-4-one 18 and azetidin-2-one 19, respectively. The newly synthesized compounds were tested for their anti-inflammatory, analgesic and anticonvulsant activities. Depending on the obtained results, the newly synthesized compounds possess significant anti-inflammatory, analgesic and anticonvulsant activities.

  19. Synthesis and Spectroscopic Studies of Mixed Ligand Complexes of Pt(II and Pd(II with Ethyl-α-Isonitrosoacetoacetate and Dienes

    Anita Krishankant Taksande


    Full Text Available The mixed ligand complexes of the kind [M(L1 (L2] where M= Pt(II, Pd(II.L1 = primary ligand ethyl-α-isonitrosoacetoacetate derived from reaction between ethyl acetoacetate, acetic acid and sodium nitrite and L2=secondary ligand para-phenyldiamine (PPD are synthesized. All the prepared complexes were identified and confirmed by elemental analysis, molar conductance measurements, and infrared electronic absorption. Their complexes has been made based on elemental analysis, molar conductivity, UV-Vis, FT-IR and 1HNMR spectroscopy and magnetic moment measurements as well as thermal analysis (TGA and DTA. The elemental analysis information recommends that the stoichiometry of the complexes to be 1:2:1. The molar conductance measurements of the complexes indicate their non-electrolytic nature. The infrared spectral information showed the coordination sites of the free ligand with the central metal particle. The electronic absorption spectral information disclosed the existence of an octahedral geometry for Pt(II and Pd(II complexes. DOI: 

  20. Synthesis and intravenous infusion into the rat of glyceryl bisacetoacetate, 1-acetoacetamido-2, 3-propane diol, and partially reduced glucosyl pentaacetoacetate.

    Birkhahn, R H; Clemens, R J; Hubbs, J C


    The efficacy of parenteral nutrition could be improved by finding a more effective energy source. Esters of short-chain fatty acids have exhibited some promise as alternatives to glucose. The present study reports on two new esters and one amide, each containing acetoacetate as the organic acid. The three compounds: glyceryl bisacetoacetate, N-2',3'-dihydroxypropyl-3-oxo-butanamide (1-acetoacetamido-2,3-propane diol), and partially reduced glucosyl pentaacetoacetate, were synthesized and then continuously infused into rats for 7 d. The infusion rate provided 50% of the rats' estimated metabolic energy requirements, and rats were fed with a reduced-energy oral diet that provided the remaining 50% of energy plus adequate protein. Rat groups for each compound were: (1) experimental-compound-infused and ad libitum-fed, (2) isoenergetic glucose-infused and pairfed, and (3) saline infused and pair-fed. Body-weight changes, N losses and N retention were measured daily. All rats died from partially reduced glucosyl pentaacetoacetate infusion at 100% and 50% of the intended rate. Rats infused with 1-acetoacetamido-2,3-propane diol failed to gain weight and to increase the plasma ketone-body concentration. Glyceryl bisacetoacetate produced hyperketonaemia, and weight gain and N variables that were similar to those for glucose-infused rats. It was concluded that only glyceryl bisacetoacetate would make a satisfactory parenteral nutrient.

  1. [Effects of allelochemical EMA isolated from Phragmites communis on algal cell membrane lipid and ultrastructure].

    Li, Feng-min; Hu, Hong-ying; Chong, Yun-xiao; Men, Yu-jie; Guo, Mei-ting


    In order to reveal the antialgal mechanisms of allelochemicals, effects of the allelochemical eathyl-2-methyl acetoacetate (EMA) on cell membrane lipid and ultrastructure of Chlorella pyrenoidosa, Microcystis aeruginosa and Chlorella vulagaris were studied in this paper. The lipid fatty acids of the algal membrane were isolated following the Bligh and Dye method and quantified by gas chromatograph/mass spectrometry. The ultrastructure of algal cells was observed with TEM. The results showed that EMA increased the contents of linolenic acid and linolic acid with increment of 14%, while decreased the content of myristic acid and cetylic acid in C. pyrenoidosa, membrane. The content of unsaturated fatty acids C18:1 and C18:2 increased 12% and 10% in M. aeruginosa with the addition of EMA, while the content of saturated fatty acids C18:0 and C16:0 decreased. EMA showed no significant change in the fatty acid composition in C. vulagaris under the experiment condition. EMA broke off cell wall of C. pyrenoidosa and M. aeruginosa. EMA damaged the cell membrane and the inclusion of algal cell leaked out. Nuclear and mitochondrial structure was damaged with the addition of EMA. EMA showed no significant change in the ultrastructure of C. vulgaris.

  2. Regulated release of Ca2+ from respiring mitochondria by Ca2+/2H+ antiport.

    Fiskum, G; Lehninger, A L


    Simultaneous measurements of oxygen consumption and transmembrane transport of Ca2+, H+, and phosphate show that the efflux of Ca2+ from respiring tightly coupled rat liver mitochondria takes place by an electroneutral Ca2+/2H+ antiport process that is ruthenium red-insensitive and that is regulated by the oxidation-reduction state of the mitochondrial pyridine nucleotides. When mitochondrial pyridine nucleotides are kept in a reduced steady state, the efflux of Ca2+ is inhibited; when they are in an oxidized state, Ca2+ efflux is activated. These processes were demonstrated by allowing phosphate-depleted mitochondria respiring on succinate in the presence of rotenone to take up Ca2+ from the medium. Upon subsequent addition of ruthenium red to block Ca2+ transport via the electrophoretic influx pathway, and acetoacetate, to bring mitochondrial pyridine nucleotides into the oxidized state, Ca2+ efflux and H+ influx ensued. The observed H+ influx/Ca2+ efflux ratio was close to the value 2.0 predicted for the operation of an electrically neutral Ca2+/2H+ antiport process.

  3. Reduction of ketones and alkyl iodides by SmI(2) and Sm(II)-HMPA complexes. Rate and mechanistic studies.

    Prasad, E; Flowers, Robert A


    The effect of HMPA on the electron transfer (ET) rate of samarium diiodide reduction reactions in THF was analyzed for a series of ketones (2-butanone, methyl acetoacetate, and N,N-dimethylacetoacetamide) and alkyl iodides (1-iodobutane and 2-iodobutane) with stopped flow spectrophotometric studies. Activation parameters for the ET processes were determined by temperature-dependence studies over a range of 30-50 degrees C. The ET rate constants and the activation parameters obtained for the above systems in the presence of different equivalents of HMPA were compared to understand the mechanism of action of HMPA on various substrates. The results obtained from these studies indicate that coordination or chelation is possible in the transition state geometry for SmI(2)/ketone systems even in the presence of the sterically demanding ligand HMPA. After the addition of 4 equiv of HMPA the ET rate and activation parameters for ketone reduction by Sm is unaffected by further HMPA addition while a linear dependence of ET rate on the equivalents of HMPA was found in the SmI(2)/alkyl iodide system. The results of these studies are consistent with an inner-sphere-type ET for the reduction of ketones by SmI(2) (and SmI(2)[bond]HMPA complexes) and an outer-sphere-type ET for the reduction of alkyl iodides by SmI(2) or SmI(2)[bond]HMPA complexes.

  4. Palladium nanoparticles bonded to two-dimensional iron oxide graphene nanosheets: a synergistic and highly reusable catalyst for the Tsuji-Trost reaction in water and air.

    Liu, Jian; Huo, Xing; Li, Tianrong; Yang, Zhengyin; Xi, Pinxian; Wang, Zhiyi; Wang, Baodui


    Low cost, high activity and selectivity, convenient separation, and increased reusability are the main requirements for noble-metal-nanocatalyst-catalyzed reactions. Despite tremendous efforts, developing noble-metal nanocatalysts to meet the above requirements remains a significant challenge. Here we present a general strategy for the preparation of strongly coupled Fe(3)O(4) and palladium nanoparticles (PdNPs) to graphene sheets by employing polyethyleneimine as the coupling linker. Transmission electron microscopic images show that Pd and Fe(3)O(4) nanoparticles are highly dispersed on the graphene surface, and the mean particle size of Pd is around 3 nm. This nanocatalyst exhibits synergistic catalysis by Pd nanoparticles supported on reduced graphene oxide (rGO) and a tertiary amine of polyethyleneimine (Pd/Fe(3)O(4)/PEI/rGO) for the Tsuji-Trost reaction in water and air. For example, the reaction of ethyl acetoacetate with allyl ethyl carbonate afforded the allylated product in more than 99 % isolated yield, and the turnover frequency reached 2200 h(-1). The yield of allylated products was 66 % for Pd/rGO without polyethyleneimine. The catalyst could be readily recycled by a magnet and reused more than 30 times without appreciable loss of activity. In addition, only about 7.5 % of Pd species leached off after 20 cycles, thus rendering this catalyst safer for the environment.

  5. 1H NMR-based metabolite profiling of plasma in a rat model of chronic kidney disease.

    Ju-Ae Kim

    Full Text Available Chronic kidney disease (CKD is characterized by the gradual loss of the kidney function to excrete wastes and fluids from the blood. (1H NMR-based metabolomics was exploited to investigate the altered metabolic pattern in rats with CKD induced by surgical reduction of the renal mass (i.e., 5/6 nephrectomy (5/6 Nx, particularly for identifying specific metabolic biomarkers associated with early of CKD. Plasma metabolite profiling was performed in CKD rats (at 4- or 8-weeks after 5/6 Nx compared to sham-operated rats. Principle components analysis (PCA, partial least squares-discriminant analysis (PLS-DA and orthogonal partial least squares-discriminant analysis (OPLS-DA score plots showed a significant separation between the groups. The resulting metabolic profiles demonstrated significantly increased plasma levels of organic anions, including citrate, β-hydroxybutyrate, lactate, acetate, acetoacetate, and formate in CKD. Moreover, levels of alanine, glutamine, and glutamate were significantly higher. These changes were likely to be associated with complicated metabolic acidosis in CKD for counteracting systemic metabolic acidosis or increased protein catabolism from muscle. In contrast, levels of VLDL/LDL (CH2n and N-acetylglycoproteins were decreased. Taken together, the observed changes of plasma metabolite profiles in CKD rats provide insights into the disturbed metabolism in early phase of CKD, in particular for the altered metabolism of acid-base and/or amino acids.

  6. Enzymatic method for determining ketone body ratio in arterial blood.

    Uno, S; Takehiro, O; Tabata, R; Ozawa, K


    We have developed a new, sensitive, and rapid method for measuring the ketone body concentration in arterial blood and determining the arterial blood ketone body ratio. The procedure involves the sequential use of the enzymes 3-hydroxybutyrate dehydrogenase (3-HBDH; EC and NADH oxidase, followed by a color-generating reaction with the hydrogen peroxide produced by the oxidase reaction. The amount of oxidized chromogen produced is proportional to the 3-hydroxybutyrate (3-HBA) concentration. The acetoacetate (AcAc) concentration is obtained after complete conversion of the AcAc to 3-HBA, in the presence of 3-HBDH. The total 3-HBA concentration is measured and then subtracted from the total ketone body concentration to give the AcAc concentration. This procedure may be applied to plasma samples and the absorbance change measured with an automated chemistry analyzer. Ketone body concentration may be determined over the range 0 to 400 mumol/L. The analysis takes approximately 12 min and requires only 30 microL of plasma.

  7. The Ketone Body, β-Hydroxybutyrate Stimulates the Autophagic Flux and Prevents Neuronal Death Induced by Glucose Deprivation in Cortical Cultured Neurons.

    Camberos-Luna, Lucy; Gerónimo-Olvera, Cristian; Montiel, Teresa; Rincon-Heredia, Ruth; Massieu, Lourdes


    Glucose is the major energy substrate in brain, however, during ketogenesis induced by starvation or prolonged hypoglycemia, the ketone bodies (KB), acetoacetate and β-hydroxybutyrate (BHB) can substitute for glucose. KB improve neuronal survival in diverse injury models, but the mechanisms by which KB prevent neuronal damage are still not well understood. In the present study we have investigated whether protection by the D isomer of BHB (D-BHB) against neuronal death induced by glucose deprivation (GD), is related to autophagy. Autophagy is a lysosomal-dependent degradation process activated during nutritional stress, which leads to the digestion of damaged proteins and organelles providing energy for cell survival. Results show that autophagy is activated in cortical cultured neurons during GD, as indicated by the increase in the levels of the lipidated form of the microtubule associated protein light chain 3 (LC3-II), and the number of autophagic vesicles. At early phases of glucose reintroduction (GR), the levels of p62 declined suggesting that the degradation of the autophagolysosomal content takes place at this time. In cultures exposed to GD and GR in the presence of D-BHB, the levels of LC3-II and p62 rapidly declined and remained low during GR, suggesting that the KB stimulates the autophagic flux preventing autophagosome accumulation and improving neuronal survival.

  8. Protection of hypoglycemia-induced neuronal death by β-hydroxybutyrate involves the preservation of energy levels and decreased production of reactive oxygen species.

    Julio-Amilpas, Alberto; Montiel, Teresa; Soto-Tinoco, Eva; Gerónimo-Olvera, Cristian; Massieu, Lourdes


    Glucose is the main energy substrate in brain but in certain circumstances such as prolonged fasting and the suckling period alternative substrates can be used such as the ketone bodies (KB), beta-hydroxybutyrate (BHB), and acetoacetate. It has been shown that KB prevent neuronal death induced during energy limiting conditions and excitotoxicity. The protective effect of KB has been mainly attributed to the improvement of mitochondrial function. In the present study, we have investigated the protective effect of D-BHB against neuronal death induced by severe noncoma hypoglycemia in the rat in vivo and by glucose deprivation (GD) in cortical cultures. Results show that systemic administration of D-BHB reduces reactive oxygen species (ROS) production in distinct cortical areas and subregions of the hippocampus and efficiently prevents neuronal death in the cortex of hypoglycemic animals. In vitro results show that D-BHB stimulates ATP production and reduces ROS levels, while the nonphysiologic isomer of BHB, L-BHB, has no effect on energy production but reduces ROS levels. Data suggest that protection by BHB, not only results from its metabolic action but is also related to its capability to reduce ROS, rendering this KB as a suitable candidate for the treatment of ischemic and traumatic injury.

  9. Classification of brain tumor extracts by high resolution ¹H MRS using partial least squares discriminant analysis

    A.V. Faria


    Full Text Available High resolution proton nuclear magnetic resonance spectroscopy (¹H MRS can be used to detect biochemical changes in vitro caused by distinct pathologies. It can reveal distinct metabolic profiles of brain tumors although the accurate analysis and classification of different spectra remains a challenge. In this study, the pattern recognition method partial least squares discriminant analysis (PLS-DA was used to classify 11.7 T ¹H MRS spectra of brain tissue extracts from patients with brain tumors into four classes (high-grade neuroglial, low-grade neuroglial, non-neuroglial, and metastasis and a group of control brain tissue. PLS-DA revealed 9 metabolites as the most important in group differentiation: γ-aminobutyric acid, acetoacetate, alanine, creatine, glutamate/glutamine, glycine, myo-inositol, N-acetylaspartate, and choline compounds. Leave-one-out cross-validation showed that PLS-DA was efficient in group characterization. The metabolic patterns detected can be explained on the basis of previous multimodal studies of tumor metabolism and are consistent with neoplastic cell abnormalities possibly related to high turnover, resistance to apoptosis, osmotic stress and tumor tendency to use alternative energetic pathways such as glycolysis and ketogenesis.

  10. Effect of Chelating Agents on the Stability of Nano-TiO2 Sol Particles for Sol-Gel Coating.

    Maeng, Wan Young; Yoo, Mi


    Agglomeration of sol particles in a titanium alkoxide (tetrabutyl orthotitanate (TBOT), > 97%) solution during the hydrolysis and condensation steps makes the sol solution difficult to use for synthesizing homogeneous sol-gel coating. Here, we have investigated the effect of stabilizing agents (acetic acid and ethyl acetoacetate (EAcAc)) on the agglomeration of Ti alkoxide particles during hydrolysis and condensation in order to determine the optimized conditions for controlling the precipitation of TiO2 particles. The study was conducted at R(AC) ([acetic acid]/[TBOT]) = 0.1-5 and R(EAcAc)([EAcAc]/[TBOT]) = 0.05-0.65. We also studied the effects of a basic catalyst ethanolamine (ETA), water, and HCl on sol stability. The chelating ligands in the precursor sol were analyzed with FT-IR. The coating properties were examined by focused ion beam. The stabilizing agents (acetic acid and EAcAc) significantly influenced the agglomeration and precipitation of TBOT precursor particles during hydrolysis. As R(AC) and R(EAcAc) increased, the agglomeration remarkably decreased. The stability of the sol with acetic acid and EAcAc arises from the coordination of the chelating ligand to TBOT that hinders hydrolysis and condensation. A uniform fine coating (thickness: 30 nm) on stainless steel was obtained by using an optimized sol with R(AC) = 0.5 and R(EAcAc) = 0.65.

  11. Ionic liquid-based dispersive liquid-liquid microextraction for the determination of formaldehyde in wastewaters and detergents.

    Arvand, Majid; Bozorgzadeh, Elahe; Shariati, Shahab; Zanjanchi, Mohammad Ali


    Spectrophotometry in combination with ionic liquid-based dispersive liquid-liquid microextraction (DLLME) was applied for the extraction and determination of formaldehyde in real samples. The method is based on the reaction of formaldehyde with methyl acetoacetate in the presence of ammonia. The variation in the absorbance of the reaction product was measured at 375 nm. An appropriate mixture of ethanol (disperser solvent) and ionic liquid, 1-hexyl-3-methylimidazoliumhexafluoro-phosphate [C(6)MIM][PF(6)] (extraction solvent) was rapidly injected into a water sample containing formaldehyde. After extraction, sedimented phase was analyzed by spectrophotometry. Under the optimum conditions, the calibration graph was linear in the range of 0.1-20 ng mL(-1) with the detection limit of 0.02 ng mL(-1) and limit of quantification of 0.08 ng mL(-1) for formaldehyde. The relative standard deviation (RSD%, n = 5) for the extraction and determination of 0.8 ng mL(-1) of formaldehyde in the aqueous samples was 2.5%. The results showed that DLLME is a very simple, rapid, sensitive, and efficient analytical method for the determination of trace amounts of formaldehyde in wastewaters and detergents, and suitable results were obtained.

  12. Microwave Assisted Synthesis and Unusual Coupling of Some Novel Pyrido[3,2-f][1,4]thiazepines

    Mohamed M. Youssef


    Full Text Available 3-Amino-3-thioxopropanamide (1 reacted with ethyl acetoacetate to form 6-hydroxy-4-methyl-2-thioxo-2,3-dihydropyridine-3-carboxamide (2, which reacted with α-haloketones 3 to produce 2,3-disubstituted-8-hydroxy-6-methyl-2H,5H-pyrido[3,2-f]-[1,4]thiazepin-5-ones 4a-c. Benzoylation of 4c led to the formation of the dibenzoate derivative 9. Compounds 4a-c could be prepared stepwise through the formation of S-alkylated derivatives 10a-c. Compounds 2, 4a-c, 9 and 10a-c were prepared using microwave as a source of heat, and gave better yields in shorter times than those achieved by traditional methods. Coupling of 4a-c with arenediazonium chlorides proceeded unusually to give the 6-hydroxy-4-methyl-2-(arylazothieno[2,3-b]pyridin-3(2H-one ring contraction products 14. Structures of the newly synthesized compounds were proven by spectral and chemical methods.

  13. [Method selected for the determination of creatinine in plasma or serum. Choice of optimal conditions of measurement].

    Labbé, D; Vassault, A; Cherruau, B; Baltassat, P; Bonète, R; Carroger, G; Costantini, A; Guérin, S; Houot, O; Lacour, B; Nicolas, A; Thioulouse, E; Trépo, D


    The method selected by the SFBC (Société française de biologie clinique) is derived from the colorimetric reaction of creatinine with alkaline picrate, measured kinetically, without any pretreatment step. The key parameters of the reaction determining the quality of the results are studied, with special regard to samples including known interferents. The aims of the study were to gain an optimal analytical sensitivity and to reduce main interferences (acetoacetate, bilirubine, glucose, protein) which plague the Jaffé reaction, through a comprehensive study of the reagents, of their concentrations and of the analytical procedures. The selected concentrations (in the test) are: 150 mmol/L sodium hydroxide, 10 mmol/L picric acid and 2 g/L sodium dodecyl sulfate. Ten millilitres of a BRIJ solution (30% volvol) are added to the reagent. The operating procedures are as follow: sample ratio 0.07 to 0.08; wavelength 505 to 510 nm; temperature 37 degrees C; incubation of the specimen with the alkaline reagent 5 mn (at least), before starting the reaction with picric acid. A seric calibrator is recommended. The first measurement is taken 20 to 40 s after starting the reaction. Total measurement time is 120 to 150 seconds.

  14. Ketosis leads to increased methylglyoxal production on the Atkins diet.

    Beisswenger, Benjamin G K; Delucia, Elizabeth M; Lapoint, Nancy; Sanford, Rebecca J; Beisswenger, Paul J


    In the popular and widely used Atkins diet, the body burns fat as its main fuel. This process produces ketosis and hence increased levels of beta-hydroxybutyrate (BOB) acetoacetate (AcAc) and its by-products acetone and acetol. These products are potential precursors of the glycotoxin methylglyoxal. Since methylglyoxal and its byproducts are recognized as a significant cause of blood vessel and tissue damage, we measured methylglyoxal, acetone, and acetol in subjects on the Atkins diet. We found that by 14-28 days, methylghyoxal levels rose 1.67-fold (P = 0.039) and acetol and acetone levels increased 2.7- and 6.12-fold, respectively (P = 0.012 and 0.028). Samples from subjects with ketosis showed even greater increases in methylglyoxal (2.12-fold), as well as acetol and acetone, which increased 4.19- and 7.9-fold, respectively; while no changes were seen in samples from noncompliant, nonketotic subjects. The increase in methylglyoxal implies that potential tissue and vascular damage can occur on the Atkins diet and should be considered when choosing a weight-loss program.

  15. Effects of a single glucocorticoid injection on propylene glycol-treated cows with clinical ketosis.

    van der Drift, Saskia G A; Houweling, Martin; Bouman, Marina; Koets, Ad P; Tielens, Aloysius G M; Nielen, Mirjam; Jorritsma, Ruurd


    This study investigated the metabolic effects of glucocorticoids when administered to propylene glycol-treated cows with clinical ketosis. Clinical ketosis was defined by depressed feed intake and milk production, and a maximal score for acetoacetate in urine. All cows received 250 mL oral propylene glycol twice daily for 3 days and were randomly assigned to a single intramuscular injection with sterile isotonic saline solution (n = 14) or dexamethasone-21-isonicotinate (n = 17). Metabolic blood variables were monitored for 6 days and adipose tissue variables for 3 days. β-Hydroxybutyrate (BHBA) concentrations in blood decreased in all cows during treatment, but were lower in glucocorticoid-treated cows. Cows treated with glucocorticoids had higher plasma glucose and insulin concentrations, whereas concentrations of non-esterified fatty acids, 3-methylhistidine and growth hormone were unaffected. mRNA expression of hormone-sensitive lipase, BHBA receptor and peroxisome proliferator-activated receptor type γ in adipose tissue was not affected. This shows that lipolytic effects do not appear to be important in ketotic cows when glucocorticoids are combined with PG. Plasma 3-methyl histidine concentrations were similar in both groups, suggesting that glucocorticoids did not increase muscle breakdown and that the greater rise in plasma glucose in glucocorticoid-treated cows may not be due to increased supply of glucogenic amino acids from muscle.

  16. Alternative energy production pathways in Taenia crassiceps cysticerci in vitro exposed to a benzimidazole derivative (RCB20).

    Fraga, Carolina Miguel; Da Costa, Tatiane Luiza; De Castro, Ana Maria; Reynoso-Ducoing, Olivia; Ambrosio, Javier; Hernández-Campos, Alicia; Castillo, Rafael; Vinaud, Marina Clare


    Biochemical studies of benzimidazole derivatives are important to determine their mode of action and activity against parasites. The lack of antihelminthic alternatives to treat parasitic infections and albendazole resistance cases make the search for new antiparasitary drugs of utmost importance. The 6-chloro-5-(1-naphthyloxy)-2-(trifluoromethyl)-1H-benzimidazole (RCB20) is a benzimidazole derivative with promising effect. This study evaluated the effect of different concentrations of RCB20 in the alternative energetic pathway of in vitro Taenia crassiceps cysticerci. The parasites were in vitro exposed to 6.5 and 13 µM of RCB20 and albendazole sulfoxide (ABZSO). The quantification of acetate, acetoacetate, β-hydroxybutyrate, fumarate and propionate was performed by high-performance liquid chromatography. The quantification of urea, creatinine and total proteins was performed by spectrophotometry. The increase in β-hydroxybutyrate reflects the enhancement of the fatty acid oxidation in the treated groups. Volatile fatty acids secretion, acetate and propionate, was increased in the treated groups. The secretion mechanisms of the treated parasites were impaired due to organic acids increased concentrations in the cysticerci. It is possible to conclude that the metabolic effect on alternative energetic pathways is slightly increased in the parasites treated with RCB20 than the ones treated with ABZSO.

  17. Direct photolysis of carbonyl compounds dissolved in cloud and fog~droplets

    Epstein, S. A.; Tapavicza, E.; Furche, F.; Nizkorodov, S. A.


    Gas-phase photolysis is an important tropospheric sink for many carbonyl compounds; however the significance of direct photolysis of these compounds dissolved in cloud and fog droplets is uncertain. We develop a theoretical approach to assess the importance of aqueous photolysis for a series of carbonyls that possess carboxyl and hydroxyl functional groups by comparison with rates of other atmospheric processes. We use computationally and experimentally derived effective Henry's law constants, hydration equilibrium parameters, aqueous hydroxyl radical (OH) rate constants, and optical extinction coefficients to identify types of compounds that will (or will not) have competitive aqueous photolysis rates. We also present molecular dynamics simulations designed to estimate gas- and aqueous-phase extinction coefficients of unstudied atmospherically relevant compounds found in d-limonene and isoprene secondary organic aerosol. In addition, experiments designed to measure the photolysis rate of glyceraldehyde, an atmospherically relevant water-soluble organic compound, reveal that aqueous quantum yields are highly molecule-specific and cannot be extrapolated from measurements of structurally similar compounds. We find that only two out of the 92 carbonyl compounds investigated, pyruvic acid and acetoacetic acid, may have aqueous photolysis rates that exceed the rate of oxidation by dissolved OH. For almost all carbonyl compounds lacking α,β-conjugation that were investigated, atmospheric removal by direct photolysis in cloud and fog droplets can be neglected under typical atmospheric conditions.

  18. The citric acid-Mn III,IVO 2(birnessite) reaction. Electron transfer, complex formation, and autocatalytic feedback

    Wang, Yun; Stone, Alan T.


    Citrate released by plants, bacteria, and fungi into soils is subject to abiotic oxidation by MnO 2(birnessite), yielding 3-ketoglutarate, acetoacetate, and Mn II. Citrate loss and generation of products as a function of time all yield S-shaped curves, indicating autocatalysis. Increasing the citrate concentration decreases the induction period. The maximum rate ( rmax) along the reaction coordinate follows a Langmuir-Hinshelwood dependence on citrate concentration. Increases in pH decrease rmax and increase the induction time. Adding Mn II, Zn II, orthophosphate, or pyrophosphate at the onset of reaction decreases rmax. Mn II addition eliminates the induction period, while orthophosphate and pyrophosphate addition increase the induction period. These findings indicate that two parallel processes are responsible. The first, relatively slow process involves the oxidation of free citrate by surface-bound Mn III,IV, yielding Mn II and citrate oxidation products. The second process, which is subject to strong positive feedback, involves electron transfer from Mn II-citrate complexes to surface-bound Mn III,IV, generating Mn III-citrate and Mn II. Subsequent intramolecular electron transfer converts Mn III-citrate into Mn II and citrate oxidation products.

  19. Fat-Free Mass Changes During Ketogenic Diets and the Potential Role of Resistance Training.

    Tinsley, Grant M; Willoughby, Darryn S


    Low-carbohydrate and very-low-carbohydrate diets are often used as weight-loss strategies by exercising individuals and athletes. Very-low-carbohydrate diets can lead to a state of ketosis, in which the concentration of blood ketones (acetoacetate, 3-β-hydroxybutyrate, and acetone) increases as a result of increased fatty acid breakdown and activity of ketogenic enzymes. A potential concern of these ketogenic diets, as with other weight-loss diets, is the potential loss of fat-free mass (e.g., skeletal muscle). On examination of the literature, the majority of studies report decreases in fat-free mass in individuals following a ketogenic diet. However, some confounding factors exist, such as the use of aggressive weight-loss diets and potential concerns with fat-free mass measurement. A limited number of studies have examined combining resistance training with ketogenic diets, and further research is needed to determine whether resistance training can effectively slow or stop the loss of fat-free mass typically seen in individuals following a ketogenic diet. Mechanisms underlying the effects of a ketogenic diet on fat-free mass and the results of implementing exercise interventions in combination with this diet should also be examined.

  20. Fenofibrate induces ketone body production in melanoma and glioblastoma cells

    Maja M Grabacka


    Full Text Available Ketone bodies (beta-hydroxybutyrate, bHB, acetoacetate are mainly produced in the liver during prolonged fasting or starvation. bHB is a very efficient energy substrate for sustaining ATP production in peripheral tissues; importantly its consumption is preferred over glucose. However, the majority of malignant cells, particularly cancer cells of neuroectodermal origin such as glioblastoma, are not able to use ketone bodies as a source of energy. Here, we report a novel observation that fenofibrate, a synthetic peroxisome proliferator-activated receptor alpha (PPARa agonist, induces bHB production in melanoma and glioblastoma cells, as well as in neurospheres composed of nontransformed cells. Unexpectedly, this effect is not dependent on PPARa activity or its expression level. The fenofibrate-induced ketogenesis is accompanied by growth arrest and down-regulation of transketolase, but the NADP/NADPH and GSH/GSSG ratios remain unaffected. Our results reveal a new, intriguing aspect of cancer cell biology and highlight the benefits of fenofibrate as a supplement to both canonical and dietary (ketogenic therapeutic approaches against glioblastoma.

  1. Cytokine-mediated inhibition of ketogenesis is unrelated to nitric oxide or protein synthesis.

    Pailla, K; El-Mir, M Y; Cynober, L; Blonde-Cynober, F


    Cytokines play an important role in the lipid disturbances commonly associated with sepsis. Ketogenesis is inhibited during sepsis, and tumor necrosis factor alpha (TNF alpha) and interleukin-6 (IL-6) have been suggested to mediate this impairment, irrespective of the ketogenic substrate (fatty acid or branched chain ketoacid). However, the underlying mechanism of cytokine action is still unknown. First we investigated the possible role of the induction of nitric oxide (NO) synthesis, using rat hepatocyte monolayers. Hepatocytes were incubated for 6 h, with either alpha -ketoisocaproate (KIC) (1 mM) or oleic acid (0.5 mM) in the presence or absence of TNF alpha (25 microg/L) and IL-6 (15 microg/L). In some experiments, cells were incubated with NO synthase (NOS) inhibitors. The ketone body (beta -hydroxybutyrate and acetoacetate) production and nitrite production were measured in the incubation medium. Our results indicated no involvement of nitric oxide in the inhibitory action of cytokines on ketogenesis. Secondly, we showed that cycloheximide (10(-4)M) did not counteract the cytokine-mediated ketogenesis decrease; hence, the effects of cytokines on ketogenesis are not protein synthesis-dependent. The cytokine-mediated inhibition of ketogenesis is therefore unrelated to either NO production or protein synthesis.

  2. Zonation of gluconeogenesis, ketogenesis and intracellular pH in livers from normal and diabetic ketoacidotic rats: evidence for intralobular redistribution of metabolic events in ketoacidosis.

    Burns, S P; Cohen, R D; Iles, R A; Bailey, R A; Desai, M; Germain, J P; Going, T C


    The intralobular distribution of metabolism was examined in the livers from rats with severe diabetic ketoacidosis (DKA), perfused at pH 6.8, and compared with that in livers from normal starved animals perfused at either pH 7.4 or 6.8. With lactate and palmitate as substrates, the perivenous uptake of periportally synthesized glucose seen in normal livers at pH 7.4 was abolished during DKA; indeed, gluconeogenesis was most active in the perivenous region. Whereas in normal livers perfused at pH 7.4 the periportal region showed a markedly elevated intracellular pH (pH(i)) compared with the perivenous zone, this distribution of pH(i) was reversed in DKA, with an intermediate distribution in normal livers perfused at pH 6. 8. 3-Hydroxybutyrate was generated throughout the lobule. Some acetoacetate generated periportally was converted to 3-hydroxybutyrate more perivenously. A steep gradient of oxygen uptake along the radius of the lobule was apparent in all three groups; oxygen uptake was greatly decreased perivenously despite adequate oxygen supply. These findings are consistent with our previous observations of the lobular co-location of high pH(i) and gluconeogenesis, and might offer an explanation of how high gluconeogenic rates can continue in spite of severe systemic acidosis in DKA. The findings provide direct evidence for a marked redistribution of intralobular metabolism in DKA.

  3. 3-Hydroxy-3-methylglutaryl CoA lyase (HL): Mouse and human HL gene (HMGCL) cloning and detection of large gene deletions in two unrelated HL-deficient patients

    Wang, S.P.; Robert, M.F.; Mitchell, G.A. [Hopital Sainte-Justine, Quebec (Canada)] [and others


    3-hydroxy-3-methylglutaryl CoA lyase (HL, EC catalyzes the cleavage of 3-hydroxy-3-methylglutaryl CoA to acetoacetic acid and acetyl CoA, the final reaction of both ketogenesis and leucine catabolism. Autosomal-recessive HL deficiency in humans results in episodes of hypoketotic hypoglycemia and coma. Using a mouse HL cDNA as a probe, we isolated a clone containing the full-length mouse HL gene that spans about 15 kb of mouse chromosome 4 and contains nine exons. The promoter region of the mouse HL gene contains elements characteristic of a housekeeping gene: a CpG island containing multiple Sp1 binding sites surrounds exon 1, and neither a TATA nor a CAAT box are present. We identified multiple transcription start sites in the mouse HL gene, 35 to 9 bases upstream of the translation start codon. We also isolated two human HL genomic clones that include HL exons 2 to 9 within 18 kb. The mouse and human HL genes (HGMW-approved symbol HMGCL) are highly homologous, with identical locations of intron-exon junctions. By genomic Southern blot analysis and exonic PCR, was found 2 of 33 HL-deficient probands to be homozygous for large deletions in the HL gene. 26 refs., 4 figs., 2 tabs.

  4. The biochemistry of ketogenesis and its role in weight management, neurological disease and oxidative stress.

    McPherson, Peter Andrew C; McEneny, Jane


    Ketogenesis is the branch of mammalian metabolism concerned with the synthesis of ketone bodies. In this process, the small, water-soluble compounds acetoacetate, D-3-β-hydroxybutyrate and propanone are produced by the liver in response to reduced glucose availability. Although ketone bodies are always present at a low level in healthy individuals, dietary manipulation and certain pathological conditions can increase the levels of these compounds in vivo. In some instances, such as in refractory epilepsy, high levels of ketone bodies can be beneficial-in this instance, by exerting an anticonvulsant effect. Conversely, if the levels of ketones rise to supraphysiological levels, as can occur in diabetes mellitus, a state of ketoacidosis can occur, which has serious consequences for cellular function. More recently, research has identified a possible link between ketogenesis and free radical-mediated pathologies, highlighting the potential application of ketogenic diets to the treatment of conditions such as Alzheimer's disease. Overall, an understanding of ketone body metabolism and its links to human disease may prove to be vital in developing new regimens for the treatment of human disease.

  5. Reciprocal effects of dietary sesamin on ketogenesis and triacylglycerol secretion by the rat liver.

    Fukuda, N; Miyagi, C; Zhang, L; Jayasooriya, A P; Sakono, M; Yamamoto, K; Ide, T; Sugano, M


    The effects of dietary sesamin (a mixture of sesamin and episesamin, 1:1, w/w) on ketone body production and lipid secretion were studied in isolated perfused liver from rats given sesamin. Feeding sesamin at the dietary level of 0.2% from 14 to 16 d resulted in an enlargement of liver weight. Ketone body production was significantly elevated in the livers perfused with oleic acid in comparison with those perfused without an exogenous-free fatty acid, and sesamin feeding caused a stimulation of ketone body production, especially when exogenous oleic acid was provided. On the other hand, the ratio of beta-hydroxybutyrate to acetoacetate, an index of mitochondrial redox potential, tended to increase in the livers perfused with oleic acid compared with those without fatty acid, thought it was consistently lowered by dietary sesamin. The cumulative secretion of triacylglycerol, but not of cholesterol, by the livers from sesamin-fed rats was decreased markedly, especially when exogenous oleic acid was provided, suggesting an inverse relationship between the rates of ketogenesis and triacylglycerol secretion. These results suggest that dietary sesamin exerts its hypotriglyceridemic effect at least in part through an enhanced metabolism of exogenous-free fatty acid to oxidation at the expense of esterification in rat liver.

  6. Intraoperative infusion of acetated Ringer solution containing glucose and ionized magnesium reduces ketogenesis and maintains serum magnesium.

    Yokoyama, Takeshi; Suwa, Kunio; Yamasaki, Fumiyasu; Yokoyama, Reiko; Yamashita, Koichi; Sellden, Eva


    The effect of glucose infusion during surgery on glucose metabolism has not been investigated sufficiently. We, therefore, examined the effect after the infusion of 1% glucose acetated Ringer solution containing Mg2+ during surgery on ketogenesis and serum Mg2+ concentrations. Patients, classified as ASA I-II, age 51-80 years, were randomly assigned to receive infusion of acetated Ringer solution. The G/Mg group received infusion with 1% glucose, Na+ 140mEq/L, Mg2+ 2 mEq/L, and the C group received infusion with glucose free solution containing Na+ 130 mEq/L without Mg2+. Both solutions were infused at a rate of 25 mL/kg for the first hour, and main-tained at 4 mL/kg/hr thereafter. Blood samples were collected three times: before infusion and at 1 hour and 4 hours after the start of infusion. Electrolytes and glucose metabolism were evaluated at each sampling. After rapid infusion, blood glucose level significantly increased to 170+/-19mg/dL in the G/Mg group, but it returned to close to baseline after 4 hours and serum ketone bodies did not increase during infusion. In the C group, however, blood glucose never increased beyond 110 mg/dL, but both acetoacetic and hydroxybutyric acids increased significantly at the third measurement.

  7. Inborn errors of ketogenesis and ketone body utilization.

    Sass, Jörn Oliver


    Ketone bodies acetoacetate and 3-hydroxy-n-butyric acid are metabolites derived from fatty acids and ketogenic amino acids such as leucine. They are mainly produced in the liver via reactions catalyzed by the ketogenic enzymes mitochondrial 3-hydroxy-3-methylglutary-coenzyme A synthase and 3-hydroxy-3-methylglutary-coenzyme A lyase. After prolonged starvation, ketone bodies can provide up to two-thirds of the brain's energy requirements. The rate-limiting enzyme of ketone body utilization (ketolysis) is succinyl-coenzyme A:3-oxoacid coenzyme A transferase. The subsequent step of ketolysis is catalyzed by 2-methylactoacetyl-coenzyme A thiolase, which is also involved in isoleucine catabolism. Inborn errors of metabolism affecting those four enzymes are presented and discussed in the context of differential diagnoses. While disorders of ketogenesis can present with hypoketotic hypoglycemia, inborn errors of ketolysis are characterized by metabolic decompensations with ketoacidosis. If those diseases are considered early and appropriate treatment is initiated without delay, patients with inborn errors of ketone body metabolism often have a good clinical outcome.

  8. Interrelations between C4 ketogenesis, C5 ketogenesis, and anaplerosis in the perfused rat liver.

    Deng, Shuang; Zhang, Guo-Fang; Kasumov, Takhar; Roe, Charles R; Brunengraber, Henri


    We investigated the interrelations between C(4) ketogenesis (production of beta-hydroxybutyrate + acetoacetate), C(5) ketogenesis (production of beta-hydroxypentanoate + beta-ketopentanoate), and anaplerosis in isolated rat livers perfused with (13)C-labeled octanoate, heptanoate, or propionate. Mass isotopomer analysis of C(4) and C(5) ketone bodies and of related acyl-CoA esters reveal that C(4) and C(5) ketogenesis share the same pool of acetyl-CoA. Although the uptake of octanoate and heptanoate by the liver are similar, the rate of C(5) ketogenesis from heptanoate is much lower than the rate of C(4) ketogenesis from octanoate. This results from the channeling of the propionyl moiety of heptanoate into anaplerosis of the citric acid cycle. C(5) ketogenesis from propionate is virtually nil because acetoacyl-CoA thiolase does not favor the formation of beta-ketopentanoyl-CoA from propionyl-CoA and acetyl-CoA. Anaplerosis and gluconeogenesis from heptanoate are inhibited by octanoate. The data have implications for the design of diets for the treatment of long chain fatty acid oxidation disorders, such as the triheptanoin-based diet.

  9. Ketone body metabolism and its defects.

    Fukao, Toshiyuki; Mitchell, Grant; Sass, Jörn Oliver; Hori, Tomohiro; Orii, Kenji; Aoyama, Yuka


    Acetoacetate (AcAc) and 3-hydroxybutyrate (3HB), the two main ketone bodies of humans, are important vectors of energy transport from the liver to extrahepatic tissues, especially during fasting, when glucose supply is low. Blood total ketone body (TKB) levels should be evaluated in the context of clinical history, such as fasting time and ketogenic stresses. Blood TKB should also be evaluated in parallel with blood glucose and free fatty acids (FFA). The FFA/TKB ratio is especially useful for evaluation of ketone body metabolism. Defects in ketogenesis include mitochondrial HMG-CoA synthase (mHS) deficiency and HMG-CoA lyase (HL) deficiency. mHS deficiency should be considered in non-ketotic hypoglycemia if a fatty acid beta-oxidation defect is suspected, but cannot be confirmed. Patients with HL deficiency can develop hypoglycemic crises and neurological symptoms even in adolescents and adults. Succinyl-CoA-3-oxoacid CoA transferase (SCOT) deficiency and beta-ketothiolase (T2) deficiency are two defects in ketolysis. Permanent ketosis is pathognomonic for SCOT deficiency. However, patients with "mild" SCOT mutations may have nonketotic periods. T2-deficient patients with "mild" mutations may have normal blood acylcarnitine profiles even in ketoacidotic crises. T2 deficient patients cannot be detected in a reliable manner by newborn screening using acylcarnitines. We review recent data on clinical presentation, metabolite profiles and the course of these diseases in adults, including in pregnancy.

  10. 云杉红翅小蠹信息素Seudenone和(±)-Seudenol的简便合成%A Facile Synthesis of Seudenone and Racemic Seudenol

    殷彩霞; 高竹林; 刘复初; 李霁良; 张甦


    Seudenone (3-methylcyclohex-2-en-1-one) is an antiaggregation pheromone of Douglas fir beetle,Dendroctonus pseudotsugae.Seudenol (3-methyl cyclohex-2-en-1-ol) is an aggregation pheromone of Douglas fir beetle,Dendroctonus pseudotsugae.The synthesis employed ethyl acetoacetate and paraformaldehyde as starting materials.The route of synthesis was carried out by 3 steps:condensation,hydrolysis and decaxboxylation,and regioselective reduction.%3-甲基环己-2-烯-1-酮(Seudenone)是云杉红翅小蠹的抑制信息素,3-甲基环己-2-烯-1-醇(±-Seudenol)却是一种聚集信息素.本合成利用乙酰乙酸乙酯和多聚甲醛为原料,通过缩合、水解脱羧和区域选择性还原三步反应合成了两个所需产物.

  11. L-carnitine: effect of intravenous administration on fuel homeostasis in normal subjects and home-parenteral-nutrition patients with low plasma carnitine concentrations.

    Bowyer, B A; Fleming, C R; Haymond, M W; Miles, J M


    We studied the effects of intravenous L-carnitine on the metabolism of fatty acids, ketone bodies, glucose, and branched-chain amino acids in four normal volunteers and four patients on long-term home parenteral nutrition (HPN) with low plasma carnitine concentrations. Substrate kinetics were determined by use of [1-14C]palmitate, [3,4-13C2]-acetoacetate, [6,6-2H2]glucose, and [5,5,5-2H3]leucine before and during a 3-h intravenous infusion of L-carnitine. HPN patients were restudied after 1 mo of nightly intravenous carnitine administration. HPN patients tolerated the short-term fast well, exhibiting neither hypoglycemia nor hypoketonemia. Intravenous carnitine had no effect on rates of fatty acid oxidation, ketone body production, glucose production, or leucine kinetics in either group. Routine addition of carnitine to the HPN regimen does not appear to be necessary. The failure of L-carnitine administration to have discernable effects on intermediary metabolism in normal volunteers casts doubt on its role in the treatment of a variety of medical conditions.

  12. Ketone-glucose interaction in fed, fasted, and fasted-infected sheep.

    Radcliffe, A G; Wolfe, R R; Colpoys, M F; Muhlbacher, F; Wilmore, D W


    Ketosis following starvation was suppressed by hindlimb infection in seven fasted sheep. Glucose production determined following the primed constant infusion of [6-3H(N)]glucose was elevated in the fasted-infected animals (9.50 +/- 1.11 mmol X kg-1 X min-1 (mean +/- SE) versus fasted controls (5.56 +/- 2.2). To determine if the ketonemia following sepsis contributed to the increased glucogenesis associated with catabolic disorder, glucose production and arterial substrates were measured before and after infusion of sodium-DL-beta-hydroxybutyrate (beta-OHB, 20 mumol X kg-1 X min-1) in fed, fasted, and fasted-infected animals. Following 3 h of beta-OHB infusion in the awake conditioned animals, beta-OHB and acetoacetate blood concentrations more than doubled. With infusion, blood glucose and alanine concentrations decreased in the fed and fasted sheep but not in the fasted-infected group. Glucose production fell significantly from 10.11 +/- 1.33 to 8.44 +/- 1.05 in the fed animals and from 5.05 +/- 0.28 to 4.11 +/- 0.33 in the fasted group. Glucose production was unaffected by beta-OHB infusion in the fasted-infected animals (9.50 +/- 1.83 vs. 9.11 +/- 1.44). The accelerated rate of glucose production in sheep following infection is not a consequence of the hypoketonemic state associated with sepsis.

  13. 3-Amino-8-hydroxy-4-imino-6-methyl-5-phenyl-4,5-dihydro-3H-chromeno [2,3-d ]pyrimidine: An Effecient Key Precursor for Novel Synthesis of Some Interesting Triazines and Triazepines as Potential Anti-Tumor Agents

    Sobhi M. Gomha


    Full Text Available A number of interesting heterocycles were prepared through interaction of the intermediate 3-amino-8-hydroxy-4-imino-6-methyl-5-phenyl-4,5-dihydro-3H-chromeno-[2,3-d]pyrimidine (1 and reagents such as hydrazonyl halides 2 to furnish triazine derivatives 4a–l. Reaction of 1 with phenacyl bromide afforded compound 5. Moreover, the title compound 1 was subjected to condensation with active methylene compounds (ethyl acetoacetate and ethyl benzoylacetate to give triazipinones 8a,b. The condensation with aromatic aldehydes afforded either the triazole derivatives 10a–d or Schiff base 11. In addition, the behaviour of compound 1 towards activated unsaturated compounds namely dimethyl acetylene dicarboxylate and ethoxymethylenemalonitrile was studied and it was found to furnish the triazine 13 and triazepine derivative 15, respectively. Combination of title compound 1 with chlorinated active methylene compounds delivered the triazine derivatives 18a–c. Reaction of 1 with chloroacetonitrile furnished compound 20. The structures of the products were elucidated based on their microanalyses and spectroscopic data. Finally, the antitumor activity of the new compounds 4a and 8a against human breast cell MCF-7 line and liver carcinoma cell line HepG2 were recorded.

  14. Synthesis of new 2-amino-4H-pyran-3,5-dicarboxylate derivatives using nanocrystalline M{sup II}Zr{sub 4}(PO{sub 4}){sub 6} ceramics as reusable and robust catalysts under microwave irradiation

    Safaei-Ghomi, Javad, E-mail: [University of Kashan, Department of Organic Chemistry, Faculty of Chemistry (Iran, Islamic Republic of); Javidan, Abdollah [Imam Hossein University, Nano-Research Center (Iran, Islamic Republic of); Ziarati, Abolfazl [University of Tehran, School of Chemistry, College of Science (Iran, Islamic Republic of); Shahbazi-Alavi, Hossein [University of Kashan, Department of Organic Chemistry, Faculty of Chemistry (Iran, Islamic Republic of)


    In the present paper, we report the successful synthesis of nanocrystalline M{sup II}Zr{sub 4}(PO{sub 4}){sub 6} ceramics (M: Mn, Ni, Fe, Co). These nano-structures were characterized by X-ray diffraction, scanning electron microscopy, and vibrating sample magnetometer (VSM). Size of nano-structures was in the range of 20–150 nm. Nano-M{sup II}Zr{sub 4}(PO{sub 4}){sub 6} as an efficient and green catalyst has been used for the preparation of 2-amino-4H-pyran-3,5-dicarboxylate derivatives by the three-component condensation reaction of ethyl cyanoacetate, ethyl acetoacetate, and various aromatic aldehydes under microwave irradiation. Extraordinarily, the best results were obtained using MnZr{sub 4}(PO{sub 4}){sub 6} nanocrystallines as an efficient catalyst. This method provides several advantages including easy work-up, excellent yields, short reaction times, using of microwave as green method, recoverability of the catalyst, and little catalyst loading.

  15. Metabolic and hormonal changes during aerobic exercise in distance runners.

    Fernández-Pastor, V J; Ruiz, M; Diego-Acosta, A M; Avila, C; García, J C; Pérez, F; Guirado, F; Noguer, N


    A group of long-distance runners is studied in order to clarify aspects concerning neuroendocrine mechanisms regulating organic adaptation to maximum effort, with special interest in the function of the growth hormone in fat metabolism and the possible use of ketone bodies as an alternative source of energy. A test is designed on a treadmill with a gradient of 3% and progressive increases in speed of 2 Km/h every 10 min, starting at 6 Km/h, and continuing until exhaustion. Masks are worn to enable the breath by breath measurement of expired gases and the subjects are monitored electrocardiographically using V5. For blood sample collection an antecubital vein is catheterized with a system enabling the replacement of the blood volume extracted by means of perfusion with physiological saline solution, and the increasing concentration of hormones in the blood is evaluated. The results obtained, indicate that epinephrine as well as GH hormones increase significatively from 20 min of exercise in runners promoting changes from carbohydrates to lipids as fuels to carry out exercise. The concomitant variations in energy substrates support the former hypothesis of work. Moreover, the muscle could employ acetylCoA originating from acetoacetate as an alternative metabolic source of fuel during maximum effort.

  16. High blood ketone body concentration in type 2 non-insulin dependent diabetic patients.

    Avogaro, A; Crepaldi, C; Miola, M; Maran, A; Pengo, V; Tiengo, A; Del Prato, S


    To assess the metabolic disturbances, and, in particular, the occurrence of high blood ketone body concentration in post-absorptive Type 2 (non-insulin-dependent) diabetic patients as compared to a matched normal population, a study was carried out in a group of 78 Type 2 diabetic outpatients matched for age and sex and in 78 normal individuals. In all subjects we measured HbA1c, and fasting levels of glucose, FFA, lactate, pyruvate, glycerol, alanine, 3-hydroxybutyrate, acetoacetate, uric acid, total cholesterol, triglycerides, creatinine, growth hormone, cortisol, glucagon, free insulin, and C-peptide. Multistix strips were used for urine ketone determination. As expected HbA1c, and plasma glucose were higher in Type 2 diabetics. This was associated with multiple metabolic disturbances as shown by higher circulating concentrations of FFA, glycerol and gluconeogenic precursors. Similarly, blood levels of ketones (351 +/- 29 vs 159 +/- 15 umol/l; P growth hormone. Plasma glucagon levels were higher in Type 2 diabetics. Blood ketone body levels were directly correlated with both plasma glucose and FFA concentrations. These observations clearly show that Type 2 diabetes is a pathologic condition characterised by multiple metabolic disturbances which are fully apparent in the basal state. Furthermore, we emphasise that Type 2 diabetic patients, though not insulin deficient, may present a significant increase in their fasting levels of ketone bodies.

  17. [Difference of acute ketone body metabolism between insulin-dependent diabetic and non-insulin-dependent diabetic individuals].

    Mayumi, K; Suzuki, S; Takuma, T; Gomi, Y; Kondo, Y; Sakamaki, T; Kokei, S; Inoue, T; Iino, S


    The difference in the acute metabolic change in ketone bodies between patients with insulin-dependent diabetes mellitus (IDDM) and non-insulin-dependent diabetes mellitus (NIDDM) was investigated in this study. The subjects employed were 7 patients with IDDM losing residual insulin secretion and 7 patients with NIDDM matched to the former patients for age, body mass index, duration of diabetes, daily insulin dosage, fasting plasma glucose and HbA1c. Blood samples were drawn at 3A.M. and 7A.M. on the same day, and plasma glucose, acetoacetic acid (AcAc), 3-beta-hydroxybutylic acid (3-OHBA), free fatty acid (FFA), glycerol, cortisol and growth hormone (GH) concentrations were determined. Plasma total ketone bodies (AcAc and 3-OHBA), 3-OHBA and FFA concentrations at 7A.M. were significantly higher in the patients with IDDM than in those with NIDDM (p ketone bodies, AcAc and 3-OHBA concentrations were also more significantly elevated in the patients with IDDM than in those with NIDDM. It was observed that the ratio of 3-OHBA was more than 2.0 in all of the patients with IDDM and less than 2.0 in all of the patients with NIDDM, the difference being significant with p < 0.001.(ABSTRACT TRUNCATED AT 250 WORDS)

  18. The effects of changes in plasma nonesterified fatty acid levels on oxidative metabolism during moderate exercise in patients with non-insulin-dependent diabetes mellitus.

    Akanji, A O; Osifo, E; Kirk, M; Hockaday, T D


    Blood levels of intermediary metabolites were measured and indirect calorimetry was performed in 10 otherwise healthy, non-insulin-dependent diabetic (NIDDM) patients before, during, and after 30 minutes of moderate exercise on three occasions in random order at weekly intervals with (1) heparin treatment to increase preexercise plasma nonesterified fatty acid (NEFA) levels (HEPARIN); (2) acipimox, a nicotinic acid analogue, to reduce preexercise plasma NEFA levels (ACIPIMOX); and (3) no manipulation of preexercise plasma NEFA levels (NIL). With ACIPIMOX, preexercise blood levels were significantly reduced for NEFAs and glycerol (P ketone bodies during either NIL or HEPARIN increased rapidly postexercise by approximately 90% to 110% for both acetoacetate and 3-hydroxybutyrate (both P growth hormone (hGH) and, perhaps, noradrenaline levels were highest both during and after exercise. The respiratory quotient (RQ) was highest with ACIPIMOX (P < .05 for exercise and postexercise periods compared with the other two treatments), which, compared with NIL, reduced fat oxidation by 27% and 60% and increased carbohydrate oxidation by 29% and 74% during and after exercise, respectively (all P < .05). These changes in substrate oxidation due to ACIPIMOX were almost opposite to those observed with HEPARIN.(ABSTRACT TRUNCATED AT 250 WORDS)

  19. NADH-generating substrates reduce peroxyl radical toxicity in RL-34 cells.

    Antosiewicz, J; Spodnik, J H; Teranishi, M; Herman-Antosiewicz, A; Kurono, Ch; Soji, T; Woźniak, M; Borkowska, A; Wakabayashi, T


    There is general agreement that oxidative stress may induce apoptotic and necrotic cell death. Recently it has been shown that NADH can be considered an important antioxidant as it reacts with peroxyl and alkoxyl radicals under in vitro conditions. Therefore, in the present study we hypothesized that an increase in intracellular NADH using specific substrates will protect RL-34 cells against cytotoxicity of 2'-azobis (2-amidinopropane) dihydrochloride (AAPH), which is a peroxyl radical generating compound. Cells treated for 24 hours with 6.0 mM AAPH were severely damaged: mitochondria were vacuolated, and the level of free radicals significantly increased. Both apoptotic and necrotic cells were detected (11.1% and 11.4%, respectively) even after 5 hours of treatment. Pretreatment of the cells with substrates which increase the intracellular level of NADH, such as lactate, beta-hydroxybutyrate, and ethanol, distinctly inhibited AAPH-induced reactive oxygen species (ROS) formation and cell death. On the other hand, acetoacetate (AcA), which decrease the intracellular level of NADH, had opposite effects. Interestingly, NADH-generating substrates augment, while AcA reduced superoxide radical formation induced by AAPH. These results may suggest that although NADH generating substrates may exert some deleterious effects within a cell by inducing reductive stress, they diminish alkoxyl or peroxyl radical cytotoxicity. The protection is associated with a decrease in ROS formation measured by dichlorofluorescein, but with an increase in superoxide radical formation.

  20. Anti-Diabetic Activity and Metabolic Changes Induced by Andrographis paniculata Plant Extract in Obese Diabetic Rats.

    Akhtar, Muhammad Tayyab; Bin Mohd Sarib, Mohamad Syakir; Ismail, Intan Safinar; Abas, Faridah; Ismail, Amin; Lajis, Nordin Hj; Shaari, Khozirah


    Andrographis paniculata is an annual herb and widely cultivated in Southeast Asian countries for its medicinal use. In recent investigations, A. paniculata was found to be effective against Type 1 diabetes mellitus (Type 1 DM). Here, we used a non-genetic out-bred Sprague-Dawley rat model to test the antidiabetic activity of A. paniculata against Type 2 diabetes mellitus (Type 2 DM). Proton Nuclear Magnetic Resonance (¹H-NMR) spectroscopy in combination with multivariate data analyses was used to evaluate the A. paniculata and metformin induced metabolic effects on the obese and obese-diabetic (obdb) rat models. Compared to the normal rats, high levels of creatinine, lactate, and allantoin were found in the urine of obese rats, whereas, obese-diabetic rats were marked by high glucose, choline and taurine levels, and low lactate, formate, creatinine, citrate, 2-oxoglutarate, succinate, dimethylamine, acetoacetate, acetate, allantoin and hippurate levels. Treatment of A. paniculata leaf water extract was found to be quite effective in restoring the disturbed metabolic profile of obdb rats back towards normal conditions. Thisstudy shows the anti-diabetic potential of A. paniculata plant extract and strengthens the idea of using this plant against the diabetes. Further classical genetic methods and state of the art molecular techniques could provide insights into the molecular mechanisms involved in the pathogenesis of diabetes mellitus and anti-diabetic effects of A. paniculata water extract.

  1. Regulation of Ca2+ release from mitochondria by the oxidation-reduction state of pyridine nucleotides.

    Lehninger, A L; Vercesi, A; Bababunmi, E A


    Mitochondria from normal rat liver and heart, and also Ehrlich tumor cells, respiring on succinate as energy source in the presence of rotenone (to prevent net electron flow to oxygen from the endogenous pyridine nucleotides), rapidly take up Ca(2+) and retain it so long as the pyridine nucleotides are kept in the reduced state. When acetoacetate is added to bring the pyridine nucleotides into a more oxidized state, Ca(2+) is released to the medium. A subsequent addition of a reductant of the pyridine nucleotides such as beta-hydroxybutyrate, glutamate, or isocitrate causes reuptake of the released Ca(2+). Successive cycles of Ca(2+) release and uptake can be induced by shifting the redox state of the pyridine nucleotides to more oxidized and more reduced states, respectively. Similar observations were made when succinate oxidation was replaced as energy source by ascorbate oxidation or by the hydrolysis of ATP. These and other observations form the basis of a hypothesis for feedback regulation of Ca(2+)-dependent substrate- or energy-mobilizing enzymatic reactions by the uptake or release of mitochondrial Ca(2+), mediated by the cytosolic phosphate potential and the ATP-dependent reduction of mitochondrial pyridine nucleotides by reversal of electron transport.

  2. Synthesis, in vitro anticancer and antimicrobial evaluation of novel substituted dihydropyrimidines

    K Rana


    Full Text Available A series of 1,4-dihydropyrimidine derivatives 3(a-t were prepared from Biginelli reactions by using ethyl acetoacetate, substituted benzaldehyde and thiourea in the presence of piperidine and ethanol. The compounds 3(a-t were reacted with dimethylsulphate, diethylsulphate, butyl bromide and benzyl chloride to give the new series of compounds 4(a-t. The structures of the newly synthesized compounds 4(a-t were established by IR, 1 H NMR, Mass spectra and elemental analysis. The synthesized compounds were evaluated for their in vitro anticancer activity by using Sulforhodamine B assay method against the growth of four humans cancer cell lines, antibacterial activity against Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa, Escherichia coli and for antifungal activity against Candida albicans and Aspergillus niger. The compounds exhibited good anticancer activity and moderate antibacterial and antifungal activities. Compounds 4b, 4c, 4d, 4g, 4i, 4n, 4o, 4q and 4s showed significant anticancer activity when compared with the doxorubicin as a standard reference drug.

  3. Effects of feeding propylene glycol on dry matter intake, lactation performance, energy balance and blood metabolites in early lactation dairy cows.

    Liu, Q; Wang, C; Yang, W Z; Zhang, W W; Yang, X M; He, D C; Dong, K H; Huang, Y X


    The objectives of this study were to evaluate effects of feeding propylene glycol (PG) on feed intake, milk yield and milk composition, blood metabolites and energy balance in Holstein dairy cows from 1 to 63 days in milk. Thirty-two multiparous cows, blocked by lactation number, previous 305-day milk production and expected calving date, were arranged into four groups in a randomized block design. Treatments were: control, low PG, medium PG and high PG with 0, 150, 300 and 450 ml PG per cow per day, respectively. The supplement of food grade PG (0.998 g/g PG) was hand-mixed into the top one-third of the daily ration. Cows were fed ad libitum a total mixed ration consisting of forage and concentrate (50 : 50, dry matter basis). Feed intake, milk yield and milk components were not affected (P > 0.05) by PG supplementation. Overall, body weight (BW) loss tended (P < 0.08) to be linearly reduced, and energy status was linearly improved with increasing PG supplementation. Concentrations of glucose in plasma were higher for cows fed PG relative to control (55.6 v. 58.9 mg/dl) and linearly increased (P < 0.01) with increasing PG supplementation. Plasma concentrations of non-esterified fatty acids and beta-hydroxybutyrate were linearly increased, but urine acetoacetate concentration was quadratically changed with the highest for control diet and the lowest for 450 ml/day of PG. These results indicated that supplementation of PG in the early lactating cow diets had minimal effects on feed intake and milk production, but may potentially reduce contents of milk fat and milk protein. Supplementation of early lactating dairy cow diets with PG is beneficial in terms of improving energy status and reducing BW loss.

  4. Escherichia coli genome-wide promoter analysis: Identification of additional AtoC binding target elements

    Kolisis Fragiskos N


    Full Text Available Abstract Background Studies on bacterial signal transduction systems have revealed complex networks of functional interactions, where the response regulators play a pivotal role. The AtoSC system of E. coli activates the expression of atoDAEB operon genes, and the subsequent catabolism of short-chain fatty acids, upon acetoacetate induction. Transcriptome and phenotypic analyses suggested that atoSC is also involved in several other cellular activities, although we have recently reported a palindromic repeat within the atoDAEB promoter as the single, cis-regulatory binding site of the AtoC response regulator. In this work, we used a computational approach to explore the presence of yet unidentified AtoC binding sites within other parts of the E. coli genome. Results Through the implementation of a computational de novo motif detection workflow, a set of candidate motifs was generated, representing putative AtoC binding targets within the E. coli genome. In order to assess the biological relevance of the motifs and to select for experimental validation of those sequences related robustly with distinct cellular functions, we implemented a novel approach that applies Gene Ontology Term Analysis to the motif hits and selected those that were qualified through this procedure. The computational results were validated using Chromatin Immunoprecipitation assays to assess the in vivo binding of AtoC to the predicted sites. This process verified twenty-two additional AtoC binding sites, located not only within intergenic regions, but also within gene-encoding sequences. Conclusions This study, by tracing a number of putative AtoC binding sites, has indicated an AtoC-related cross-regulatory function. This highlights the significance of computational genome-wide approaches in elucidating complex patterns of bacterial cell regulation.

  5. Facile Synthesis, Characterization, and Antimicrobial Evaluation of Novel Heterocycles, Schiff Bases, and N-Nucleosides Bearing Phthalazine Moiety.

    Azab, Mohamed Emad; Rizk, Sameh Ahmed; Mahmoud, Naglaa Fawzy


    The present work describes convenient synthesis of the novel Schiff bases 5a and b by reacting phthalazinones 4a and b with 4-methoxybenzaldehyde Reaction of the Schiff bases with phenylisothiocyanate afforded diazetidine derivatives 7a and b. Also, compounds 4a and b reacted with 2-bromoglucoside tetraacetate giving peracetylated N-glycosides 6a and b, which were deacetylated to afford N-glycosylated phthalazinones 8a and b. On the other hand, when compound 3 was treated with POCl3/PCl5 and/or ethyl chloroacetate, chlorophthalazine and ethyl acetate derivatives 9 and 10 were obtained, respectively. Hydrazinolysis of compounds 9 and 10 produced the hydrazino and hydrazide derivatives 11 and 12, respectively. When compound 11 reacted with 2-furanaldehyde, acetic anhydride, and/or carbon disulphide, it gave compounds 13-15, respectively. Treatment of the hydrazide 12 with aromatic aldehydes, acetic anhydride, ethyl acetoacetate, acetyl acetone, ammonium thiocyanate, and/or phthalic anhydride furnished compounds 17-21. Meanwhile, reacting Schiff base 22 with the chlorophthalazine derivative 9 produced compound 23, which on treatment with furoyl chloride produced compound 24. The structures of the novel compounds were confirmed by IR, (1)H-NMR, (13)C-NMR, MS, and elemental analysis. The newly synthesized compounds were tested against Bacillus subtilis and Staphylococcus aureus as Gram-positive bacteria, Escherichia coli and Pseudomonas aurignosa as Gram-negative bacteria, and Candida albicans and Aspergillus niger as fungi strains. Compounds 5a and b, 23, and 24 showed greater antimicrobial activity than the stranded compounds, suggesting that they could be considered as promising antimicrobial agents.

  6. Advances in Synthesis and Aromatization of Hantzsch 1,4-Dihydropyridine Derivatives%Hantzsch 1,4-二氢吡啶衍生物的合成及其芳香化研究进展



      综述了Hantzsch 1,4-二氢吡啶衍生物的合成及其芳香化研究进展.Hantzsch 1,4-二氢吡啶衍生物的合成方法有:(1)乙酰乙酸乙酯、芳香醛和氨或伯胺反应;(2)1,3-二酮化合物、芳香醛和氨或伯胺反应;(3)烯胺化合物和芳香醛反应;(4)1,5-二酮与氨反应;(5) a,b-不饱和酮与烯胺或酮和氨反应等.在合成技术上有固相合成和微波辅助合成等.Hantzsch 1,4-二氢吡啶芳香化方法有:光化学反应、电化学反应、脱氢反应、氢转移反应等,本文主要对脱氢反应研究进展进行综述.%  The main methods of synthesis and aromatization of Hantzsch 1,4-dihydropyridine derivatives was reviewed in this paper. The former included the reactions of ethyl acetoacetate and aromaticaldehyde with ammonia or primary amine, the reactions of 1,3-diketone compounds and aromaticaldehyde with ammonia or primary amine, the reactions of enamine compounds with aromaticaldehyde, the reactions of 1,5-diketone with amine, the reactions of a,b-unsaturated ketone with enamine or ketone and amine. The synthetic technologies included solid-phase synthesis and microwave-assistant synthesis. The latter included photochemical reactions, electrochemical reactions, dehydrogenation reactions, hydrogen transfer reactions.

  7. The ketone metabolite β-hydroxybutyrate blocks NLRP3 inflammasome-mediated inflammatory disease.

    Youm, Yun-Hee; Nguyen, Kim Y; Grant, Ryan W; Goldberg, Emily L; Bodogai, Monica; Kim, Dongin; D'Agostino, Dominic; Planavsky, Noah; Lupfer, Christopher; Kanneganti, Thirumala D; Kang, Seokwon; Horvath, Tamas L; Fahmy, Tarek M; Crawford, Peter A; Biragyn, Arya; Alnemri, Emad; Dixit, Vishwa Deep


    The ketone bodies β-hydroxybutyrate (BHB) and acetoacetate (AcAc) support mammalian survival during states of energy deficit by serving as alternative sources of ATP. BHB levels are elevated by starvation, caloric restriction, high-intensity exercise, or the low-carbohydrate ketogenic diet. Prolonged fasting reduces inflammation; however, the impact that ketones and other alternative metabolic fuels produced during energy deficits have on the innate immune response is unknown. We report that BHB, but neither AcAc nor the structurally related short-chain fatty acids butyrate and acetate, suppresses activation of the NLRP3 inflammasome in response to urate crystals, ATP and lipotoxic fatty acids. BHB did not inhibit caspase-1 activation in response to pathogens that activate the NLR family, CARD domain containing 4 (NLRC4) or absent in melanoma 2 (AIM2) inflammasome and did not affect non-canonical caspase-11, inflammasome activation. Mechanistically, BHB inhibits the NLRP3 inflammasome by preventing K(+) efflux and reducing ASC oligomerization and speck formation. The inhibitory effects of BHB on NLRP3 are not dependent on chirality or starvation-regulated mechanisms like AMP-activated protein kinase (AMPK), reactive oxygen species (ROS), autophagy or glycolytic inhibition. BHB blocks the NLRP3 inflammasome without undergoing oxidation in the TCA cycle, and independently of uncoupling protein-2 (UCP2), sirtuin-2 (SIRT2), the G protein-coupled receptor GPR109A or hydrocaboxylic acid receptor 2 (HCAR2). BHB reduces NLRP3 inflammasome-mediated interleukin (IL)-1β and IL-18 production in human monocytes. In vivo, BHB or a ketogenic diet attenuates caspase-1 activation and IL-1β secretion in mouse models of NLRP3-mediated diseases such as Muckle-Wells syndrome, familial cold autoinflammatory syndrome and urate crystal-induced peritonitis. Our findings suggest that the anti-inflammatory effects of caloric restriction or ketogenic diets may be linked to BHB

  8. 一种新型Hsp90抑制剂的合成及其抑制活性%Synthesis of Novel Hsp90 Inhibotor and Its Inhibition

    张晓颖; 张杰; 李伟; 王小龙


    对氨基苯甲酸经重氮化后再与2-氯乙酰乙酸乙醅反应制得(Z)4-[2-(1-氯-2-乙氧基-2-羰亚甲基)肼基]苯甲酸(1);1与二甲基环己二酮成环后再与对羟基环己胺完成酰胺化反应合成了一种新型Hs90抑制剂——N-(4-羟基环己基)-4-[1-(6,6-二甲基-4-氧-3-乙酰氧基-4,5,6,7-四氢吲唑)]苯甲酰胺(3),其结构经1 H NMR表征.荧光偏振法研究结果表明3对Hsp90α具有明显的抑制活性.%(Z)-4-[2-(l-chloro-2-ethoxy-2-oxoethylidene)hydrazinyl]benzoic acid(l) was prepared from p-aminobenzoic acid by diazotization, then reacted with ethyl 2-chloro-acetoacetate. A novel Hsp90 inhibotor, ethyl l-{4-[ (4-hydroxycyclohexyl)carbamoyl]phenyl} -6,6-dimethyl-4-oxo-4,5,6, 7-tetrahydro-lH-indazole-3-carboxylate(3), was synthesized by cyclization between 1 and 5,5-dime-thyl-cyclohexane-1,3-dione, then amidation with p-hydroxyl-cyclohexylamine. The structure was characterized by ' H NMR. The result from fluorescence polarization showed that 3 exhibited good inhibition against Hsp90a.

  9. 2,4-二甲基吡咯-3,5-二羧酸二乙酯的合成--推荐一个大学有机化学实验%Synthesis of Diethyl 2,4-dimethyl-pyrrole-3,5-dicarboxylate:Recommending a University Organic Chemistry Experiment

    汪剑波; 宗乾收; 沈锦锦; 毕成; 吴晨俊


    Using Knorr pyrrole synthesis, diethyl 2,4-dimethyl-pyrrole-3,5-dicarboxylate was synthesized via one-pot method with ethyl acetoacetate and sodium nitrite as starting materials in the presence of acetic acid and zinc powder. The experiment involves the experimental operation and analyses including temperature control, reflux, recrystal ization, melting point, IR and NMR spectroscopies. The advantages of the experiment are simple raw materials and high product yield. The experiment is helpful for cultivating and improving the synthesis ability of students, as wel as in-depth understanding of heterocyclic synthesis and spectral analysis.%采用Knorr吡咯合成法,以乙酰乙酸乙酯和亚硝酸钠为原料,在醋酸和锌粉作用下采用“一锅法”得到2,4-二甲基吡咯-3,5-二羧酸二乙酯。本实验涉及到控温、回流、重结晶、熔点测定、红外光谱及核磁共振等实验操作和分析检测方法。该实验原料简单易得、产物收率高,结合波谱解析方法,可以培养和提高学生合成实验的能力,加强学生对杂环合成和波谱解析的理解与分析。

  10. Glucose tolerance of pregnant sows is related to postnatal pig mortality.

    Kemp, B; Soede, N M; Vesseur, P C; Helmond, F A; Spoorenberg, J H; Frankena, K


    An experiment was conducted on three farms to study the relationship between the results of an oral glucose tolerance test on d 104 +/- 4 of pregnancy and subsequent reproductive performance of 104 multiparous sows. After an overnight period without feed, sows were fed (3 g/kg BW.75) glucose, and blood samples were taken for analyses of glucose at -10, 10, 20, 30, 40, 50, 60 70, 80, 90, 105, and 120 min after administration of glucose. Glucose concentrations before glucose administration ranged from 2.0 to 3.6 mmol/L. Sows with lesser pretest glucose concentrations had longer (P < .01) durations of pregnancy (corrected for number of live pigs born) and heavier (P < .05) pigs at birth (corrected for farm and number of live pigs born). Peak glucose concentrations and areas under the curves (i.e., first 70 min and entire 120 min) ranged from .1 to 4.4 mmol/L, -40 to 211 (mmol/L).min, and -95 to 247 (mmol/L).min, respectively. Sows with greater (P < .01) peak concentration of glucose and greater area under the curve (first 70 min [P < .01] and entire 120 min [P < .01]) had increased pig mortality during the first 7 d after farrowing (corrected for live weight of the sows). In 83 sows, a sample of morning urine (before feeding) was collected and tested for glucose and ketone bodies. Neither glucose nor the ketone body acetoacetic acid was detected in the urine. The results indicate that sows that are less glucose-tolerant have greater pig mortality.

  11. The neuropharmacology of butyrate: The bread and butter of the microbiota-gut-brain axis?

    Stilling, Roman M; van de Wouw, Marcel; Clarke, Gerard; Stanton, Catherine; Dinan, Timothy G; Cryan, John F


    Several lines of evidence suggest that brain function and behaviour are influenced by microbial metabolites. Key products of the microbiota are short-chain fatty acids (SCFAs), including butyric acid. Butyrate is a functionally versatile molecule that is produced in the mammalian gut by fermentation of dietary fibre and is enriched in butter and other dairy products. Butyrate along with other fermentation-derived SCFAs (e.g. acetate, propionate) and the structurally related ketone bodies (e.g. acetoacetate and d-β-hydroxybutyrate) show promising effects in various diseases including obesity, diabetes, inflammatory (bowel) diseases, and colorectal cancer as well as neurological disorders. Indeed, it is clear that host energy metabolism and immune functions critically depend on butyrate as a potent regulator, highlighting butyrate as a key mediator of host-microbe crosstalk. In addition to specific receptors (GPR43/FFAR2; GPR41/FFAR3; GPR109a/HCAR2) and transporters (MCT1/SLC16A1; SMCT1/SLC5A8), its effects are mediated by utilisation as an energy source via the β-oxidation pathway and as an inhibitor of histone deacetylases (HDACs), promoting histone acetylation and stimulation of gene expression in host cells. The latter has also led to the use of butyrate as an experimental drug in models for neurological disorders ranging from depression to neurodegenerative diseases and cognitive impairment. Here we provide a critical review of the literature on butyrate and its effects on multiple aspects of host physiology with a focus on brain function and behaviour. We find fundamental differences in natural butyrate at physiological concentrations and its use as a neuropharmacological agent at rather high, supraphysiological doses in brain research. Finally, we hypothesise that butyrate and other volatile SCFAs produced by microbes may be involved in regulating the impact of the microbiome on behaviour including social communication.

  12. Effects of ketone bodies in Alzheimer's disease in relation to neural hypometabolism, β-amyloid toxicity, and astrocyte function.

    Hertz, Leif; Chen, Ye; Waagepetersen, Helle S


    Diet supplementation with ketone bodies (acetoacetate and β-hydroxybuturate) or medium-length fatty acids generating ketone bodies has consistently been found to cause modest improvement of mental function in Alzheimer's patients. It was suggested that the therapeutic effect might be more pronounced if treatment was begun at a pre-clinical stage of the disease instead of well after its manifestation. The pre-clinical stage is characterized by decade-long glucose hypometabolism in brain, but ketone body metabolism is intact even initially after disease manifestation. One reason for the impaired glucose metabolism may be early destruction of the noradrenergic brain stem nucleus, locus coeruleus, which stimulates glucose metabolism, at least in astrocytes. These glial cells are essential in Alzheimer pathogenesis. The β-amyloid peptide Aβ interferes with their cholinergic innervation, which impairs synaptic function because of diminished astrocytic glutamate release. Aβ also reduces glucose metabolism and causes hyperexcitability. Ketone bodies are similarly used against seizures, but the effectively used concentrations are so high that they must interfere with glucose metabolism and de novo synthesis of neurotransmitter glutamate, reducing neuronal glutamatergic signaling. The lower ketone body concentrations used in Alzheimer's disease may owe their effect to support of energy metabolism, but might also inhibit release of gliotransmitter glutamate. Alzheimer's disease is a panglial-neuronal disorder with long-standing brain hypometabolism, aberrations in both neuronal and astrocytic glucose metabolism, inflammation, hyperexcitability, and dementia. Relatively low doses of β-hydroxybutyrate can have an ameliorating effect on cognitive function. This could be because of metabolic supplementation or inhibition of Aβ-induced release of glutamate as gliotransmitter, which is likely to reduce hyperexcitability and inflammation. The therapeutic

  13. Metabolic conversion of dicarboxylic acids to succinate in rat liver homogenates. A stable isotope tracer study.

    Tserng, K Y; Jin, S J


    The metabolic conversion of dicarboxylic acids into succinate and other gluconeogenic intermediates in rat liver homogenates was investigated using [1,2,4-13C4]dodecanedioic acid as tracer. Isotope enrichments in 3-hydroxybutyrate, succinate, fumarate, and malate, as well as dicarboxylates (dodecanedioic, sebacic, suberic, and adipic acids) were measured with selected ion monitoring capillary column gas chromatograph-mass spectrometry. Significant enrichment in the M + 4 (four labeled carbons) ion of succinate (0.4-2.9%) was detected, unequivocally demonstrating the direct conversion of dicarboxylate into succinate. In addition, significant enrichment of the M + 2 ion of succinate was also observed. This labeled species was generated from labeled acetyl-CoA through the tricarboxylic acid cycle. The partition of acetyl-CoA into the tricarboxylic acid cycle relative to ketone body formation was higher in the beta oxidation of dicarboxylate than monocarboxylate. Therefore, in addition to the production of succinate, the beta oxidation of dodecanedioate resulted in the channeling of the acetyl-CoA produced to the tricarboxylic acid cycle instead of to acetoacetate production. The enrichments in lower chain dicarboxylates are consistent with a partial bidirectional beta oxidation of dodecanedioic acid. In addition to the expected M + 0 and M + 4 labels, significant M + 2 species were detected in suberic and adipic acids. These M + 2-labeled species were produced from the released free dicarboxylate intermediates which were then reactivated and metabolized. In these experiments, the overall succinate production was derived 4% from the direct conversion of dodecanedioic acid and 11% from the indirect route via acetyl-CoA through tricarboxylic acid.

  14. Isopropanol production with engineered Cupriavidus necator as bioproduction platform.

    Grousseau, Estelle; Lu, Jingnan; Gorret, Nathalie; Guillouet, Stéphane E; Sinskey, Anthony J


    Alleviating our society's dependence on petroleum-based chemicals has been highly emphasized due to fossil fuel shortages and increasing greenhouse gas emissions. Isopropanol is a molecule of high potential to replace some petroleum-based chemicals, which can be produced through biological platforms from renewable waste carbon streams such as carbohydrates, fatty acids, or CO2. In this study, for the first time, the heterologous expression of engineered isopropanol pathways were evaluated in a Cupriavidus necator strain Re2133, which was incapable of producing poly-3-hydroxybutyrate [P(3HB)]. These synthetic production pathways were rationally designed through codon optimization, gene placement, and gene dosage in order to efficiently divert carbon flow from P(3HB) precursors toward isopropanol. Among the constructed pathways, Re2133/pEG7c overexpressing native C. necator genes encoding a β-ketothiolase, a CoA-transferase, and codon-optimized Clostridium genes encoding an acetoacetate decarboxylase and an alcohol dehydrogenase produced up to 3.44 g l(-1) isopropanol in batch culture, from fructose as a sole carbon source, with only 0.82 g l(-1) of biomass. The intrinsic performance of this strain (maximum specific production rate 0.093 g g(-1) h(-1), yield 0.32 Cmole Cmole(-1)) corresponded to more than 60 % of the respective theoretical performance. Moreover, the overall isopropanol production yield (0.24 Cmole Cmole(-1)) and the overall specific productivity (0.044 g g(-1) h(-1)) were higher than the values reported in the literature to date for heterologously engineered isopropanol production strains in batch culture. Strain Re2133/pEG7c presents good potential for scale-up production of isopropanol from various substrates in high cell density cultures.

  15. An insight into the metabolic responses of ultra-small superparamagnetic particles of iron oxide using metabonomic analysis of biofluids

    Feng Jianghua [Department of Physics, Fujian Key Laboratory of Plasma and Magnetic Resonance, State Key Laboratory of Physical Chemistry of Solid Surfaces, Xiamen University, Xiamen, 361005 (China); Liu Huili; Zhang Limin [State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, Wuhan Institute of Physics and Mathematics, Chinese Academy of Sciences, Wuhan 430071 (China); Bhakoo, Kishore [Singapore Bioimaging Consortium, Agency for Science, Technology and Research (A-STAR) 138667 (Singapore); Lu Lehui, E-mail:, E-mail: [State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin 130022 (China)


    Ultra-small superparamagnetic particles of iron oxides (USPIO) have been developed as intravenous organ/tissue-targeted contrast agents to improve magnetic resonance imaging (MRI) in vivo. However, their potential toxicity and effects on metabolism have attracted particular attention. In the present study, uncoated and dextran-coated USPIO were investigated by analyzing both rat urine and plasma metabonomes using high-resolution NMR-based metabonomic analysis in combination with multivariate statistical analysis. The wealth of information gathered on the metabolic profiles from rat urine and plasma has revealed subtle metabolic changes in response to USPIO administration. The metabolic changes include the elevation of urinary {alpha}-hydroxy-n-valerate, o- and p-HPA, PAG, nicotinate and hippurate accompanied by decreases in the levels of urinary {alpha}-ketoglutarate, succinate, citrate, N-methylnicotinamide, NAG, DMA, allantoin and acetate following USPIO administration. The changes associated with USPIO administration included a gradual increase in plasma glucose, N-acetyl glycoprotein, saturated fatty acid, citrate, succinate, acetate, GPC, ketone bodies ({beta}-hydroxybutyrate, acetone and acetoacetate) and individual amino acids, such as phenylalanine, lysine, isoleucine, glycine, glutamine and glutamate and a gradual decrease of myo-inositol, unsaturated fatty acid and triacylglycerol. Hence USPIO administration effects are reflected in changes in a number of metabolic pathways including energy, lipid, glucose and amino acid metabolism. The size- and surface chemistry-dependent metabolic responses and possible toxicity were observed using NMR analysis of biofluids. These changes may be attributed to the disturbances of hepatic, renal and cardiac functions following USPIO administrations. The potential biotoxicity can be derived from metabonomic analysis and serum biochemistry analysis. Metabonomic strategy offers a promising approach for the detection of

  16. Acetone production with metabolically engineered strains of Acetobacterium woodii.

    Hoffmeister, Sabrina; Gerdom, Marzena; Bengelsdorf, Frank R; Linder, Sonja; Flüchter, Sebastian; Öztürk, Hatice; Blümke, Wilfried; May, Antje; Fischer, Ralf-Jörg; Bahl, Hubert; Dürre, Peter


    Expected depletion of oil and fossil resources urges the development of new alternative routes for the production of bulk chemicals and fuels beyond petroleum resources. In this study, the clostridial acetone pathway was used for the formation of acetone in the acetogenic bacterium Acetobacterium woodii. The acetone production operon (APO) containing the genes thlA (encoding thiolase A), ctfA/ctfB (encoding CoA transferase), and adc (encoding acetoacetate decarboxylase) from Clostridium acetobutylicum were cloned under the control of the thlA promoter into four vectors having different replicons for Gram-positives (pIP404, pBP1, pCB102, and pCD6). Stable replication was observed for all constructs. A. woodii [pJIR_actthlA] achieved the maximal acetone concentration under autotrophic conditions (15.2±3.4mM). Promoter sequences of the genes ackA from A. woodii and pta-ack from C. ljungdahlii were determined by primer extension (PEX) and cloned upstream of the APO. The highest acetone production in recombinant A. woodii cells was achieved using the promoters PthlA and Ppta-ack. Batch fermentations using A. woodii [pMTL84151_actthlA] in a bioreactor revealed that acetate concentration had an effect on the acetone production, due to the high Km value of the CoA transferase. In order to establish consistent acetate concentration within the bioreactor and to increase biomass, a continuous fermentation process for A. woodii was developed. Thus, acetone productivity of the strain A. woodii [pMTL84151_actthlA] was increased from 1.2mgL(-1)h(-1) in bottle fermentation to 26.4mgL(-1)h(-1) in continuous gas fermentation.

  17. High acetone-butanol-ethanol production in pH-stat co-feeding of acetate and glucose.

    Gao, Ming; Tashiro, Yukihiro; Wang, Qunhui; Sakai, Kenji; Sonomoto, Kenji


    We previously reported the metabolic analysis of butanol and acetone production from exogenous acetate by (13)C tracer experiments (Gao et al., RSC Adv., 5, 8486-8495, 2015). To clarify the influence of acetate on acetone-butanol-ethanol (ABE) production, we first performed an enzyme assay in Clostridium saccharoperbutylacetonicum N1-4. Acetate addition was found to drastically increase the activities of key enzymes involved in the acetate uptake (phosphate acetyltransferase and CoA transferase), acetone formation (acetoacetate decarboxylase), and butanol formation (butanol dehydrogenase) pathways. Subsequently, supplementation of acetate during acidogenesis and early solventogenesis resulted in a significant increase in ABE production. To establish an efficient ABE production system using acetate as a co-substrate, several shot strategies were investigated in batch culture. Batch cultures with two substrate shots without pH control produced 14.20 g/L butanol and 23.27 g/L ABE with a maximum specific butanol production rate of 0.26 g/(g h). Furthermore, pH-controlled (at pH 5.5) batch cultures with two substrate shots resulted in not only improved acetate consumption but also a further increase in ABE production. Finally, we obtained 15.13 g/L butanol and 24.37 g/L ABE at the high specific butanol production rate of 0.34 g/(g h) using pH-stat co-feeding method. Thus, in this study, we established a high ABE production system using glucose and acetate as co-substrates in a pH-stat co-feeding system with C. saccharoperbutylacetonicum N1-4.

  18. The extracellular redox state modulates mitochondrial function, gluconeogenesis, and glycogen synthesis in murine hepatocytes.

    Laura Nocito

    Full Text Available Circulating redox state changes, determined by the ratio of reduced/oxidized pairs of different metabolites, have been associated with metabolic diseases. However, the pathogenic contribution of these changes and whether they modulate normal tissue function is unclear. As alterations in hepatic gluconeogenesis and glycogen metabolism are hallmarks that characterize insulin resistance and type 2 diabetes, we tested whether imposed changes in the extracellular redox state could modulate these processes. Thus, primary hepatocytes were treated with different ratios of the following physiological extracellular redox couples: β-hydroxybutyrate (βOHB/acetoacetate (Acoc, reduced glutathione (GSH/oxidized glutathione (GSSG, and cysteine/cystine. Exposure to a more oxidized ratio via extracellular βOHB/Acoc, GSH/GSSG, and cysteine/cystine in hepatocytes from fed mice increased intracellular hydrogen peroxide without causing oxidative damage. On the other hand, addition of more reduced ratios of extracellular βOHB/Acoc led to increased NAD(PH and maximal mitochondrial respiratory capacity in hepatocytes. Greater βOHB/Acoc ratios were also associated with decreased β-oxidation, as expected with enhanced lipogenesis. In hepatocytes from fasted mice, a more extracellular reduced state of βOHB/Acoc led to increased alanine-stimulated gluconeogenesis and enhanced glycogen synthesis capacity from added glucose. Thus, we demonstrated for the first time that the extracellular redox state regulates the major metabolic functions of the liver and involves changes in intracellular NADH, hydrogen peroxide, and mitochondrial respiration. Because redox state in the blood can be communicated to all metabolically sensitive tissues, this work confirms the hypothesis that circulating redox state may be an important regulator of whole body metabolism and contribute to alterations associated with metabolic diseases.

  19. Engineering yield and rate of reductive biotransformation in Escherichia coli by partial cyclization of the pentose phosphate pathway and PTS-independent glucose transport.

    Siedler, Solvej; Bringer, Stephanie; Blank, Lars M; Bott, Michael


    Optimization of yields and productivities in reductive whole-cell biotransformations is an important issue for the industrial application of such processes. In a recent study with Escherichia coli, we analyzed the reduction of the prochiral β-ketoester methyl acetoacetate by an R-specific alcohol dehydrogenase (ADH) to the chiral hydroxy ester (R)-methyl 3-hydroxybutyrate (MHB) using glucose as substrate for the generation of NADPH. Deletion of the phosphofructokinase gene pfkA almost doubled the yield to 4.8 mol MHB per mole of glucose, and it was assumed that this effect was due to a partial cyclization of the pentose phosphate pathway (PPP). Here, this partial cyclization was confirmed by (13)C metabolic flux analysis, which revealed a negative net flux from glucose 6-phosphate to fructose 6-phosphate catalyzed by phosphoglucose isomerase. For further process optimization, the genes encoding the glucose facilitator (glf) and glucokinase (glk) of Zymomonas mobilis were overexpressed in recombinant E. coli strains carrying ADH and deletions of either pgi (phosphoglucose isomerase), or pfkA, or pfkA plus pfkB. In all cases, the glucose uptake rate was increased (30-47%), and for strains Δpgi and ΔpfkA also, the specific MHB production rate was increased by 15% and 20%, respectively. The yield of the latter two strains slightly dropped by 11% and 6%, but was still 73% and 132% higher compared to the reference strain with intact pgi and pfkA genes and expressing glf and glk. Thus, metabolic engineering strategies are presented for improving yield and rate of reductive redox biocatalysis by partial cyclization of the PPP and by increasing glucose uptake, respectively.

  20. Effects of feeding dry glycerin to early postpartum Holstein dairy cows on lactational performance and metabolic profiles.

    Chung, Y-H; Rico, D E; Martinez, C M; Cassidy, T W; Noirot, V; Ames, A; Varga, G A


    Effects of feeding a dry glycerin product (minimal 65% of food grade glycerol, dry powder) to 39 multiparous Holstein dairy cows (19 control and 20 glycerin-supplemented; lactation number = 2.2 +/- 1.3 SD) on feed intake, milk yield and composition, and blood metabolic profiles were investigated. Dry glycerin was fed at 250 g/d as a top dressing (corresponding to 162.5 g of glycerol/d) to the common lactating total mixed ration from parturition to 21 d postpartum. Individual milk was sampled from 2 consecutive milkings weekly and analyzed for components. Blood was sampled from the coccygeal vein at 4, 7, 14, and 21 (+/-0.92, pooled SD) d in milk and analyzed for urea nitrogen, glucose, insulin, nonesterified fatty acids, and beta-hydroxybutyrate. Urine was tested for the acetoacetate level weekly by using Ketostix. Average feed intake, milk yield and components, blood metabolites, and serum insulin concentrations were not affected by dry glycerin supplementation. Glycerin-supplemented cows experienced a more positive energy status (higher concentrations of plasma glucose, lower concentrations of plasma beta-hydroxybutyrate, and lower concentrations of urine ketones), which was observed during the second week of lactation, suggesting that energy availability may have been improved. This glucogenic effect of dry glycerin did not result in an increase in feed intake or milk yield during the first 3 wk of lactation, likely because of the relatively less negative energy status of cows transitioning into lactation. The tendency toward higher milk yield for glycerin-supplemented cows during wk 6 of lactation (52 vs. 46 kg/d) after the supplementation period (dry glycerin was terminated at wk 3 of lactation) suggested a potential benefit of dry glycerin on subsequent milk production, perhaps through changes in metabolism, which requires further investigation.

  1. NMR-based metabonomic study of the sub-acute toxicity of titanium dioxide nanoparticles in rats after oral administration

    Bu Qian; Lin Hongjun; Xu Youzhi; Cao Zhixing; Zhou Tian; Zhao Yinglan [State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041 (China); Yan Guangyan; Cen Xiaobo [National Chengdu Center for Safety Evaluation of Drugs, State Key Laboratory of Biotherapy, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041 (China); Deng Pengchi [Analytical and Testing Center, Sichuan University, Chengdu 610041 (China); Peng Feng [Department of Thoracic Oncology of Cancer Center and State Key Laboratory of Biotherapy, West China Hospital, West China Medical School, Sichuan University, Chengdu 610041 (China); Xue Aiqin [Institute of Bioengineering, Zhejiang Sci-Tech University Road 2, Xiasha, Hangzhou 310018 (China); Wang Yanli, E-mail: [Tianjin Children' s Hospital, Tianjin 300074 (China)


    As titanium dioxide nanoparticles (TiO{sub 2} NPs) are widely used commercially, their potential toxicity on human health has attracted particular attention. In the present study, the oral toxicological effects of TiO{sub 2} NPs (dosed at 0.16, 0.4 and 1 g kg{sup -1}, respectively) were investigated using conventional approaches and metabonomic analysis in Wistar rats. Serum chemistry, hematology and histopathology examinations were performed. The urine and serum were investigated by {sup 1}H nuclear magnetic resonance (NMR) using principal components and partial least squares discriminant analysis. The metabolic signature of urinalysis in TiO{sub 2} NP-treated rats showed increases in the levels of taurine, citrate, hippurate, histidine, trimethylamine-N-oxide (TMAO), citrulline, {alpha}-ketoglutarate, phenylacetylglycine (PAG) and acetate; moreover, decreases in the levels of lactate, betaine, methionine, threonine, pyruvate, 3-D-hydroxybutyrate (3-D-HB), choline and leucine were observed. The metabonomics analysis of serum showed increases in TMAO, choline, creatine, phosphocholine and 3-D-HB as well as decreases in glutamine, pyruvate, glutamate, acetoacetate, glutathione and methionine after TiO{sub 2} NP treatment. Aspartate aminotransferase (AST), creatine kinase (CK) and lactate dehydrogenase (LDH) were elevated and mitochondrial swelling in heart tissue was observed in TiO{sub 2} NP-treated rats. These findings indicate that disturbances in energy and amino acid metabolism and the gut microflora environment may be attributable to the slight injury to the liver and heart caused by TiO{sub 2} NPs. Moreover, the NMR-based metabolomic approach is a reliable and sensitive method to study the biochemical effects of nanomaterials.

  2. [Effects of allelochemical isolated from Phragmites communis on algal membrane permeability].

    Li, Feng-Min; Hu, Hong-Ying; Chong, Yun-Xiao; Guo, Mei-Ting; Men, Yu-Jie


    Efflux of K+, Mg2+, Ca2+ ions from algal cells as signals of cell membrane permeability, inductively coupled plasma mass spectrometry (ICP-MS) as detection method of ions, the present research investigated effects of allelochemical eathyl-2-methyl acetoacetate (EMA) isolated from Phragmites communis on cell membrane permeability of Microcystis aeruginosa, Chlorella pyrenoidosa and Chlorella vulagaris. The results showed that, when the cells were boiled for 10 min and the membrane was destroyed absolutely, the K+ efflux of M. aeruginosa and C. pyrenoidosa were 1.45 and 1.59 microg x (10(9) cell) (-1), respectively. When the concentration of EMA was 2 mg x L(-1), the K+ efflux of M. aeruginosa and C. pyrenoidosa were 1.38 and 1.40 microg x (10(9) cell)(-1), respectively. The K+ efflux of M. aeruginosa and C. pyrenoidosa reached 1.44 and 1.58 microg x (10(9) cell)(-1) while the EMA was 4 mg x L(-1). When the concentrations were 2 mg x L(-1) or 4 mg x L(-1) the K+ efflux reached more than 95% of the total ion amount in M. aeruginosa and C. pyrenoidosa cells. But when EMA concentration was 4 mg x L(-1), K+ efflux of C. vulagaris was 0.64 microg x (10(9) cell)(-1), which was only 31.5% of total K+ amount in C. vulagaris. Effects EMA on efflux of Mg2+ and Ca2+ were similar to those of K+. The results indicated that EMA destroyed the cell membrane of M. aeruginosa and C. pyrenoidosa but not C. vulagaris. This is one of the mechanisms of EMA species-selective antialgal.

  3. Taxonomic characterization and metabolic analysis of the Halomonas sp. KM-1, a highly bioplastic poly(3-hydroxybutyrate)-producing bacterium.

    Kawata, Yoshikazu; Shi, Lian-Hua; Kawasaki, Kazunori; Shigeri, Yasushi


    In a brief previous report, the gram-negative moderately halophilic bacterium, Halomonas sp. KM-1, that was isolated in our laboratory was shown to produce the bioplastic, poly(3-hydroxybutyrate) (PHB), using biodiesel waste glycerol (Kawata and Aiba, Biosci. Biotechnol. Biochem., 74, 175-177, 2010). Here, we further characterized this KM-1 strain and compared it to other Halomonas strains. Strain KM-1 was subjected to a polyphasic taxonomic study. Strain KM-1 was rod-shaped and formed colonies on a plate that were cream-beige in color, smooth, opaque, and circular with entire edges. KM-1 grew under environmental conditions of 0.1%-10% (w/v) NaCl, pH 6.5-10.5 and at temperatures between 10°C and 45°C. The G+C content of strain KM-1 was 63.9 mol%. Of the 16 Halomonas strains examined in this study, the strain KM-1 exhibited the highest production of PHB (63.6%, w/v) in SOT medium supplemented with 10% glycerol, 10.0 g/L sodium nitrate and 2.0 g/L dipotassium hydrogen phosphate. The intracellular structures within which PHB accumulated had the appearance of intracellular granules with a diameter of approximately 0.5 μm, as assessed by electron microscopy. The intra- and extra-cellular metabolites of strain KM-1 were analyzed by capillary electrophoresis mass spectrometry. In spite of the high amount of PHB stored intra-cellularly, as possible precursors for PHB only a small quantity of 3-hydroxybutyric acid and acetyl CoA, and no quantity of 3-hydroxybutyl CoA, acetoacetyl CoA and acetoacetate were detected either intra- or extra-cellularly, suggesting highly efficient conversion of these precursors to PHB.

  4. Metabolomics of Human Amniotic Fluid and Maternal Plasma during Normal Pregnancy.

    Orczyk-Pawilowicz, Magdalena; Jawien, Ewa; Deja, Stanislaw; Hirnle, Lidia; Zabek, Adam; Mlynarz, Piotr


    Metabolic profiles of amniotic fluid and maternal blood are sources of valuable information about fetus development and can be potentially useful in diagnosis of pregnancy disorders. In this study, we applied 1H NMR-based metabolic profiling to track metabolic changes occurring in amniotic fluid (AF) and plasma (PL) of healthy mothers over the course of pregnancy. AF and PL samples were collected in the 2nd (T2) and 3rd (T3) trimester, prolonged pregnancy (PP) until time of delivery (TD). A multivariate data analysis of both biofluids reviled a metabolic switch-like transition between 2nd and 3rd trimester, which was followed by metabolic stabilization throughout the rest of pregnancy probably reflecting the stabilization of fetal maturation and development. The differences were further tested using univariate statistics at α = 0.001. In plasma the progression from T2 to T3 was related to increasing levels of glycerol, choline and ketone bodies (3-hydroxybutyrate and acetoacetate) while pyruvate concentration was significantly decreased. In amniotic fluid, T2 to T3 transition was associated with decreasing levels of glucose, carnitine, amino acids (valine, leucine, isoleucine, alanine, methionine, tyrosine, and phenylalanine) and increasing levels of creatinine, succinate, pyruvate, choline, N,N-dimethylglycine and urocanate. Lactate to pyruvate ratio was decreased in AF and conversely increased in PL. The results of our study, show that metabolomics profiling can be used to better understand physiological changes of the complex interdependencies of the mother, the placenta and the fetus during pregnancy. In the future, these results might be a useful reference point for analysis of complicated pregnancies.

  5. Synthesis and conformational study of some r(2), c(4)-bis(isopropoxycarbonyl)- t(3)-aryl- c(5)-hydroxy- t(5)-methylcyclohexanones using NMR spectra

    Pandiarajan, K.; Mohan, R. T. Sabapathy; Murugavel, K.; Hema, R.


    Seven r(2), c(4)-bis(isopropoxycarbonyl)- t(3)-aryl- c(5)-hydroxy- t(5)-methylcyclohexanones 7- 13 (aryl = C 6H 5, p-ClC 6H 4, p-FC 6H 4, p-OMeC 6H 4, p-Me 2NC 6H 4, m-O 2NC 6H 4 and m-C 6H 5OC 6H 4) have been synthesized by condensing isopropyl acetoacetate with aromatic aldehydes in the presence of methylamine. For all these compounds 1H and 13C NMR spectra have been recorded. For 7, HOMOCOSY, NOESY, HSQC and HMBC spectra also have been recorded. The spectral data suggest that compounds 7- 13 exist in chair conformation with axial orientation of the hydroxy group and equatorial orientations of all the other substituents. Long range coupling is observed between OH proton and H(6a) in all cases suggesting that OH group is anti to C(5) sbnd C(6) bond. In all cases four distinct doublets are observed for the methyl protons of the two isopropyl groups. In 7 and 13 four separate, closely spaced signals are observed for the methyl carbons of the two isopropyl groups. All signals could be assigned unambiguously in the case of 7. The protons of one methyl group of the isopropoxycarbonyl group at C-4 seems to be highly shielded. This suggests that these protons lie above the plane of the aryl group at C(3). This conclusion is in accordance with the structure of 7, determined by X-ray crystallography. MOPAC calculations on 7 suggest that the chair conformation with OH group anti to C(5) sbnd C(6) bond is more stable than the chair conformation with OH bond anti to C(5) sbnd CH 3 bond by 1.5 kcal mol -1.

  6. Can Ketones Help Rescue Brain Fuel Supply in Later Life? Implications for Cognitive Health during Aging and the Treatment of Alzheimer's Disease.

    Cunnane, Stephen C; Courchesne-Loyer, Alexandre; Vandenberghe, Camille; St-Pierre, Valérie; Fortier, Mélanie; Hennebelle, Marie; Croteau, Etienne; Bocti, Christian; Fulop, Tamas; Castellano, Christian-Alexandre


    We propose that brain energy deficit is an important pre-symptomatic feature of Alzheimer's disease (AD) that requires closer attention in the development of AD therapeutics. Our rationale is fourfold: (i) Glucose uptake is lower in the frontal cortex of people >65 years-old despite cognitive scores that are normal for age. (ii) The regional deficit in brain glucose uptake is present in adults old who have genetic or lifestyle risk factors for AD but in whom cognitive decline has not yet started. Examples include young adult carriers of presenilin-1 or apolipoprotein E4, and young adults with mild insulin resistance or with a maternal family history of AD. (iii) Regional brain glucose uptake is impaired in AD and mild cognitive impairment (MCI), but brain uptake of ketones (beta-hydroxybutyrate and acetoacetate), remains the same in AD and MCI as in cognitively healthy age-matched controls. These observations point to a brain fuel deficit which appears to be specific to glucose, precedes cognitive decline associated with AD, and becomes more severe as MCI progresses toward AD. Since glucose is the brain's main fuel, we suggest that gradual brain glucose exhaustion is contributing significantly to the onset or progression of AD. (iv) Interventions that raise ketone availability to the brain improve cognitive outcomes in both MCI and AD as well as in acute experimental hypoglycemia. Ketones are the brain's main alternative fuel to glucose and brain ketone uptake is still normal in MCI and in early AD, which would help explain why ketogenic interventions improve some cognitive outcomes in MCI and AD. We suggest that the brain energy deficit needs to be overcome in order to successfully develop more effective therapeutics for AD. At present, oral ketogenic supplements are the most promising means of achieving this goal.

  7. Metabolomics provide new insights on lung cancer staging and discrimination from chronic obstructive pulmonary disease.

    Deja, Stanislaw; Porebska, Irena; Kowal, Aneta; Zabek, Adam; Barg, Wojciech; Pawelczyk, Konrad; Stanimirova, Ivana; Daszykowski, Michal; Korzeniewska, Anna; Jankowska, Renata; Mlynarz, Piotr


    Chronic obstructive pulmonary disease (COPD) and lung cancer are widespread lung diseases. Cigarette smoking is a high risk factor for both the diseases. COPD may increase the risk of developing lung cancer. Thus, it is crucial to be able to distinguish between these two pathological states, especially considering the early stages of lung cancer. Novel diagnostic and monitoring tools are required to properly determine lung cancer progression because this information directly impacts the type of the treatment prescribed. In this study, serum samples collected from 22 COPD and 77 lung cancer (TNM stages I, II, III, and IV) patients were analyzed. Then, a collection of NMR metabolic fingerprints was modeled using discriminant orthogonal partial least squares regression (OPLS-DA) and further interpreted by univariate statistics. The constructed discriminant models helped to successfully distinguish between the metabolic fingerprints of COPD and lung cancer patients (AUC training=0.972, AUC test=0.993), COPD and early lung cancer patients (AUC training=1.000, AUC test=1.000), and COPD and advanced lung cancer patients (AUC training=0.983, AUC test=1.000). Decreased acetate, citrate, and methanol levels together with the increased N-acetylated glycoproteins, leucine, lysine, mannose, choline, and lipid (CH3-(CH2)n-) levels were observed in all lung cancer patients compared with the COPD group. The evaluation of lung cancer progression was also successful using OPLS-DA (AUC training=0.811, AUC test=0.904). Based on the results, the following metabolite biomarkers may prove useful in distinguishing lung cancer states: isoleucine, acetoacetate, and creatine as well as the two NMR signals of N-acetylated glycoproteins and glycerol.

  8. Serum metabonomic analysis of protective effects of Curcuma aromatica oil on renal fibrosis rats.

    Liangcai Zhao

    Full Text Available BACKGROUND: Curcuma aromatica oil is a traditional herbal medicine demonstrating protective and anti-fibrosis activities in renal fibrosis patients. However, study of its mechanism of action is challenged by its multiple components and multiple targets that its active agent acts on. METHODOLOGY/PRINCIPAL FINDINGS: Nuclear magnetic resonance (NMR-based metabonomics combined with clinical chemistry and histopathology examination were performed to evaluate intervening effects of Curcuma aromatica oil on renal interstitial fibrosis rats induced by unilateral ureteral obstruction. The metabolite levels were compared based on integral values of serum 1H NMR spectra from rats on 3, 7, 14, and 28 days after the medicine administration. Time trajectory analysis demonstrated that metabolic profiles of the agent-treated rats were restored to control levels after 7 days of dosage. The results confirmed that the agent would be an effective anti-fibrosis medicine in a time-dependent manner, especially in early renal fibrosis stage. Targeted metabolite analysis showed that the medicine could lower levels of lipid, acetoacetate, glucose, phosphorylcholine/choline, trimethylamine oxide and raise levels of pyruvate, glycine in the serum of the rats. Serum clinical chemistry and kidney histopathology examination dovetailed well with the metabonomics data. CONCLUSIONS/SIGNIFICANCES: The results substantiated that Curcuma aromatica oil administration can ameliorate renal fibrosis symptoms by inhibiting some metabolic pathways, including lipids metabolism, glycolysis and methylamine metabolism, which are dominating targets of the agent working in vivo. This study further strengthens the novel analytical approach for evaluating the effect of traditional herbal medicine and elucidating its molecular mechanism.

  9. NMR-based metabonomic study of the sub-acute toxicity of titanium dioxide nanoparticles in rats after oral administration

    Bu, Qian; Yan, Guangyan; Deng, Pengchi; Peng, Feng; Lin, Hongjun; Xu, Youzhi; Cao, Zhixing; Zhou, Tian; Xue, Aiqin; Wang, Yanli; Cen, Xiaobo; Zhao, Ying-Lan


    As titanium dioxide nanoparticles (TiO2 NPs) are widely used commercially, their potential toxicity on human health has attracted particular attention. In the present study, the oral toxicological effects of TiO2 NPs (dosed at 0.16, 0.4 and 1 g kg - 1, respectively) were investigated using conventional approaches and metabonomic analysis in Wistar rats. Serum chemistry, hematology and histopathology examinations were performed. The urine and serum were investigated by 1H nuclear magnetic resonance (NMR) using principal components and partial least squares discriminant analysis. The metabolic signature of urinalysis in TiO2 NP-treated rats showed increases in the levels of taurine, citrate, hippurate, histidine, trimethylamine-N-oxide (TMAO), citrulline, α-ketoglutarate, phenylacetylglycine (PAG) and acetate; moreover, decreases in the levels of lactate, betaine, methionine, threonine, pyruvate, 3-D-hydroxybutyrate (3-D-HB), choline and leucine were observed. The metabonomics analysis of serum showed increases in TMAO, choline, creatine, phosphocholine and 3-D-HB as well as decreases in glutamine, pyruvate, glutamate, acetoacetate, glutathione and methionine after TiO2 NP treatment. Aspartate aminotransferase (AST), creatine kinase (CK) and lactate dehydrogenase (LDH) were elevated and mitochondrial swelling in heart tissue was observed in TiO2 NP-treated rats. These findings indicate that disturbances in energy and amino acid metabolism and the gut microflora environment may be attributable to the slight injury to the liver and heart caused by TiO2 NPs. Moreover, the NMR-based metabolomic approach is a reliable and sensitive method to study the biochemical effects of nanomaterials.

  10. Redox state and energy metabolism during liver regeneration: alterations produced by acute ethanol administration.

    Gutiérrez-Salinas, J; Miranda-Garduño, L; Trejo-Izquierdo, E; Díaz-Muñoz, M; Vidrio, S; Morales-González, J A; Hernández-Muñoz, R


    Ethanol metabolism can induce modifications in liver metabolic pathways that are tightly regulated through the availability of cellular energy and through the redox state. Since partial hepatectomy (PH)-induced liver proliferation requires an oversupply of energy for enhanced syntheses of DNA and proteins, the present study was aimed at evaluating the effect of acute ethanol administration on the PH-induced changes in cellular redox and energy potentials. Ethanol (5 g/kg body weight) was administered to control rats and to two-thirds hepatectomized rats. Quantitation of the liver content of lactate, pyruvate, beta-hydroxybutyrate, acetoacetate, and adenine nucleotides led us to estimate the cytosolic and mitochondrial redox potentials and energy parameters. Specific activities in the liver of alcohol-metabolizing enzymes also were measured in these animals. Liver regeneration had no effect on cellular energy availability, but induced a more reduced cytosolic redox state accompanied by an oxidized mitochondrial redox state during the first 48 hr of treatment; the redox state normalized thereafter. Administration of ethanol did not modify energy parameters in PH rats, but this hepatotoxin readily blocked the PH-induced changes in the cellular redox state. In addition, proliferating liver promoted decreases in the activity of alcohol dehydrogenase (ADH) and of cytochrome P4502E1 (CYP2E1); ethanol treatment prevented the PH-induced diminution of ADH activity. In summary, our data suggest that ethanol could minimize the PH-promoted metabolic adjustments mediated by redox reactions, probably leading to an ineffective preparatory event that culminates in compensatory liver growth after PH in the rat.

  11. Roles and regulation of ketogenesis in cultured astroglia and neurons under hypoxia and hypoglycemia.

    Takahashi, Shinichi; Iizumi, Takuya; Mashima, Kyoko; Abe, Takato; Suzuki, Norihiro


    Exogenous ketone bodies (KBs), acetoacetate (AA), and β-hydroxybutyrate (BHB) act as alternative energy substrates in neural cells under starvation. The present study examined the endogenous ketogenic capacity of astroglia under hypoxia with/without glucose and the possible roles of KBs in neuronal energy metabolism. Cultured neurons and astroglia were prepared from Sprague-Dawley rats. Palmitic acid (PAL) and l-carnitine (LC) were added to the assay medium. The 4- to 24-hr production of AA and BHB was measured using the cyclic thio-NADH method. (14)C-labeled acid-soluble products (KBs) and (14)CO2 produced from [1-(14)C]PAL were also measured. l-[U-(14)C]lactic acid ([(14)C]LAC), [1-(14)C]pyruvic acid ([(14)C]PYR), or β-[1-(14)C]hydroxybutyric acid ([(14)C]BHB) was used to compare the oxidative metabolism of the glycolysis end products with that of the KBs. Some cells were placed in a hypoxic chamber (1% O2). PAL and LC induced a higher production of KBs in astroglia than in neurons, while the CO2 production from PAL was less than 5% of the KB production in both astroglia and neurons. KB production in astroglia was augmented by the AMP-activated protein kinase activators, AICAR and metformin, as well as hypoxia with/without glucose. Neuronal KB production increased under hypoxia in the absence of PAL and LC. In neurons, [(14)C]LAC and [(14)C]PYR oxidation decreased after 24 hr of hypoxia, while [(14)C]BHB oxidation was preserved. Astroglia responds to ischemia in vitro by enhancing KB production, and astroglia-produced KBs derived from fatty acid might serve as a neuronal energy substrate for the tricarboxylic acid cycle instead of lactate, as pyruvate dehydrogenase is susceptible to ischemia.

  12. Quantitation of ketogenesis in periportal and pericentral regions of the liver lobule.

    Olson, M J; Thurman, R G


    A method has been devised to quantitate rates of ketogenesis (acetoacetate + beta-hydroxybutyrate production) in discrete regions of the liver lobule based on changes in NADH fluorescence. In perfused livers from fasted rats, ketogenesis was inhibited nearly completely with either 2-bromoctanoate (600 microM) or 2-tetradecylglycidic acid (25 microM). During inhibition of ketogenesis, a linear relationship (r = 0.90) was observed between decreases in NADH fluorescence detected from the liver surface and decreases in ketone body production. NADH fluorescence was monitored subsequently from individual regions of the liver lobule by placing microlight guides on periportal and pericentral regions of the liver lobule visible on the liver surface. Rates of ketogenesis in sublobular regions were calculated from regional decreases in NADH fluorescence and changes in the rate of ketone body formation by the whole liver during infusion of inhibitors. In the presence of bromoctanoate, ketogenesis was reduced 80% and local rates of ketogenesis were decreased 31 +/- 4 mumol/g/h in periportal areas and 28 +/- 3 mumol/g/h in pericentral regions. Similar results were observed with tetradecylglycidic acid. Therefore, it was concluded that submaximal rates of ketogenesis from endogenous, mainly long-chain fatty acids are nearly equal in periportal and pericentral regions of the liver lobule in liver from fasted rats. Rates of ketogenesis and NADH fluorescence were strongly correlated during fatty acid infusion. Infusion of 250 microM oleate increased NADH fluorescence maximally by 8 +/- 1% over basal values in periportal regions and 17 +/- 4% in pericentral areas. Local rates of ketogenesis, calculated from these changes in fluorescence, increased 35 +/- 6 mumol/g/h in periportal areas and 55 +/- 5 mumol/g/h in pericentral regions. Thus, oleate stimulated ketogenesis nearly 60% more in pericentral than in periportal regions of the liver lobule.

  13. Roles and Regulation of Ketogenesis in Cultured Astroglia and Neurons Under Hypoxia and Hypoglycemia

    Shinichi Takahashi


    Full Text Available Exogenous ketone bodies (KBs, acetoacetate (AA, and β-hydroxybutyrate (BHB act as alternative energy substrates in neural cells under starvation. The present study examined the endogenous ketogenic capacity of astroglia under hypoxia with/without glucose and the possible roles of KBs in neuronal energy metabolism. Cultured neurons and astroglia were prepared from Sprague-Dawley rats. Palmitic acid (PAL and l-carnitine (LC were added to the assay medium. The 4- to 24-hr production of AA and BHB was measured using the cyclic thio-NADH method. 14C-labeled acid-soluble products (KBs and 14CO2 produced from [1-14C]PAL were also measured. l-[U-14C]lactic acid ([14C]LAC, [1-14C]pyruvic acid ([14C]PYR, or β-[1-14C]hydroxybutyric acid ([14C]BHB was used to compare the oxidative metabolism of the glycolysis end products with that of the KBs. Some cells were placed in a hypoxic chamber (1% O2. PAL and LC induced a higher production of KBs in astroglia than in neurons, while the CO2 production from PAL was less than 5% of the KB production in both astroglia and neurons. KB production in astroglia was augmented by the AMP-activated protein kinase activators, AICAR and metformin, as well as hypoxia with/without glucose. Neuronal KB production increased under hypoxia in the absence of PAL and LC. In neurons, [14C]LAC and [14C]PYR oxidation decreased after 24 hr of hypoxia, while [14C]BHB oxidation was preserved. Astroglia responds to ischemia in vitro by enhancing KB production, and astroglia-produced KBs derived from fatty acid might serve as a neuronal energy substrate for the tricarboxylic acid cycle instead of lactate, as pyruvate dehydrogenase is susceptible to ischemia.

  14. Synthesis of novel furobenzopyrone derivatives and evaluation of their antimicrobial and antiinflammatory activity

    Srinivasan K


    Full Text Available Certain 4′-(4′′-substituted phenyl-4-methylfurobenzopyrones were synthesized and evaluated for antibacterial activity. Six of the synthesized compounds were also screened for their antiinflammatory activity. Substituted resorcinols were condensed with ethyl acetoacetate to afford different coumarins (2a-c. Various substituted phenacyl bromides (4a-g were prepared by the bromination of para-substituted acetophenones. The coumarins (2a-c and phenacyl bromides (4a-g were condensed to give oxoethers (5a-s. These were cyclised by using 1 M sodium hydroxide to afford the desired furobenzopyrone derivatives (FCa-s. All the compounds have been evaluated for their antibacterial activity against different strains of gram positive and gram negative bacteria. All the compounds have shown good activity against Pseudomonas aeruginosa . Compounds, 3-(4-chlorophenyl-5-methylfuro-[3,2-g][1]benzopyran-7-one, 3-(4-chlorophenyl-5,9-dimethylfuro[3,2-g][1]benzopyran-7-one and 4,5-dimethyl-3-phenylfuro[3,2-g][1]benzopyran-7-one (FCe, FCi, FCn were active against E. coli . A few compounds showed moderate activity against Bacillus subtilis also. Antiinflammatory activity of six selected compounds was also tested using the carrageenan-induced rat paw oedema method. Among them, 5-methyl-3-p-tolylfuro[3,2-g][1]benzopyran-7-one (FCg showed excellent activity. 5-Methyl-3-phenylfuro[3,2-g][1]benzopyran-7-one (FCa and 4,5-dimethyl-3-(4-nitrophenyl-furo[3,2-g][1]benzopyran-7-one (FCc showed activity comparable to that of the standard drug ibuprofen.

  15. Effect of intravenous L-carnitine on growth parameters and fat metabolism during parenteral nutrition in neonates.

    Helms, R A; Mauer, E C; Hay, W W; Christensen, M L; Storm, M C


    To determine whether intravenous carnitine can improve nutritional indices, neonates requiring parenteral nutrition were randomized into carnitine treatment (n = 23) and control (n = 20) groups. Observed plasma lipid indices, carnitine and nitrogen balances, and plasma carnitine concentrations were not different in the prestudy period. Under standardized, steady-state conditions, 0.5 g/kg Intralipid was administered intravenously over 2 hr prior to carnitine administration, after infants received 7 days of 50 mumol/kg/day, and after a second 7 days of 100 mumol/kg/day of continuous intravenous L-carnitine as part of parenteral nutrition. Triglyceride (TGY), free fatty acid (FFA), acetoacetate (AA), beta-hydroxybutyrate (BOB), and plasma carnitine concentrations were measured prior to and at 2, 4, and 6 hr after the initiation of the lipid bolus. Twenty-four-hour urine collections for nitrogen and carnitine balance were obtained on days 7 and 14. Neonates receiving carnitine had significantly greater concentrations of plasma carnitine on days 7 and 14 (p less than 0.001). Greater nitrogen (p less than 0.05) and carnitine (p less than 0.001) balances and weight gain (week 2, p less than 0.05) were found in the carnitine-supplemented group when compared with controls. On day 14, (BOB + AA)/FFA ratios were significantly higher (p less than 0.05), and peak TGY concentrations and 6-hr FFA concentrations were significantly lower (p less than 0.05) in the treatment group. Carnitine supplementation was associated with modest increases in growth and nitrogen accretion possibly by enhancing the neonate's ability to utilize exogenous fat for energy.

  16. Aging Impairs Myocardial Fatty Acid and Ketone Oxidation and Modifies Cardiac Functional and Metabolic Responses to Insulin in Mice

    Hyyti, Outi M.; Ledee, Dolena; Ning, Xue-Han; Ge, Ming; Portman, Michael A.


    Aging presumably initiates shifts in substrate oxidation mediated in part by changes in insulin sensitivity. Similar shifts occur with cardiac hypertrophy and may contribute to contractile dysfunction. We tested the hypothesis that aging modifies substrate utilization and alters insulin sensitivity in mouse heart when provided multiple substrates. In vivo cardiac function was measured with microtipped pressure transducers in the left ventricle from control (4–6 mo) and aged (22–24 mo) mice. Cardiac function was also measured in isolated working hearts along with substrate and anaplerotic fractional contributions to the citric acid cycle (CAC) by using perfusate containing 13C-labeled free fatty acids (FFA), acetoacetate, lactate, and unlabeled glucose. Stroke volume and cardiac output were diminished in aged mice in vivo, but pressure development was preserved. Systolic and diastolic functions were maintained in aged isolated hearts. Insulin prompted an increase in systolic function in aged hearts, resulting in an increase in cardiac efficiency. FFA and ketone flux were present but were markedly impaired in aged hearts. These changes in myocardial substrate utilization corresponded to alterations in circulating lipids, thyroid hormone, and reductions in protein expression for peroxisome proliferator-activated receptor (PPAR)α and pyruvate dehydrogenase kinase (PDK)4. Insulin further suppressed FFA oxidation in the aged. Insulin stimulation of anaplerosis in control hearts was absent in the aged. The aged heart shows metabolic plasticity by accessing multiple substrates to maintain function. However, fatty acid oxidation capacity is limited. Impaired insulin-stimulated anaplerosis may contribute to elevated cardiac efficiency, but may also limit response to acute stress through depletion of CAC intermediates.

  17. Metabolic effects of vasopressin infusion in the starved rat. Reversal of ketonaemia.

    Rofe, A M; Williamson, D H


    The effects of vasopressin on the metabolism of starved rats were investigated by using a constant-infusion regimen (50 pmol/kg body wt. per min, after an initial loading dose of 150 pmol/kg body wt.). 2. Blood ketone bodies decreased by 50% in 10 min, and this was accompanied by a 60% decrease in the plasma non-esterified fatty acids. 3. Blood glucose increased by 0.9 mM within 5 min and decreased to control values over the 40 min infusion. Small increases in lactate and pyruvate also occurred. 4. Plasma insulin was not increased by vasopressin infusion. 5. The net decrease in blood ketone bodies caused by vasopressin was similar when somatostatin was infused simultaneously (1 nmol/kg body wt. per min). 6. Hepatic ketone bodies were significantly decreased by vasopressin, as was the 3-hydroxybutyrate/acetoacetate ratio. A small increase in the hepatic concentration of several glycolytic intermediates also occurred. 7. Vasopressin did not decrease the ketonaemia produced by infusions of octanoate or long-chain triacylglycerol in rats that had been pre-treated with the anti-lipolytic agent 3,5-dimethylpyrazole. 8. In comparison with vasopressin, the infusion of adrenaline or glucose had much smaller effects in decreasing the ketonaemia of starvation, despite the 4-fold increase in plasma insulin, at 10 min, with the glucose infusion. 9. The primary metabolic effect of vasopressin in the starved rat appears to be that of decreased supply of non-esterified fatty acid to the liver. It is suggested that vasopressin has a direct anti-lipolytic effect in adipose tissue. PMID:6135420

  18. The origin of free brain malonate

    Riley, K.M.; Dickson, A.C.; Koeppen, A.H. (Research Service, Veterans Administration Medical Center, Albany, NY (United States))


    Rat brain contains substantial concentrations of free malonate (192 nmol/g wet weight) but origin and biological importance of the dicarboxylic acid are poorly understood. A dietary source has been excluded. A recently described malonyl-CoA decarboxylase deficiency is associated with malonic aciduria and clinical manifestations, including mental retardation. In an effort to study the metabolic origin of free malonate, several labeled acetyl-CoA precursors were administered by intracerebral injection. (2-14C)pyruvate or (1,5-14C)citrate produced radioactive glutamate but failed to label malonate. In contrast, (1-14C)acetate, (2-14C)acetate, and (1-14C)butyrate were converted to labeled glutamate and malonate after the same route of administration. The intracerebral injection of (1-14C)-beta-alanine as a precursor of malonic semialdehyde and possibly free malonate did not give rise to radioactivity in the dicarboxylate. The labeling pattern of malonic acid is compatible with the reaction sequence: acetyl-CoA----malonyl-CoA----malonate. The final step is thought to occur by transfer of the CoA-group from malonyl-CoA to succinate and/or acetoacetate. Labeling of malonate from acetate is most effective at the age of 7 days when the net concentration of the dicarboxylic acid in rat brain is still very low. At this age, butyrate was a better precursor of malonate than acetate. It is proposed that fatty acid oxidation provides the acetyl-CoA which functions as the precursor of free brain malonate. Compartmentation of malonate biosynthesis is likely because the acetyl-CoA precursors citrate and pyruvate are ineffective.

  19. Reactions of Cg10062, a cis-3-Chloroacrylic Acid Dehalogenase Homologue, with Acetylene and Allene Substrates: Evidence for a Hydration-Dependent Decarboxylation.

    Huddleston, Jamison P; Johnson, William H; Schroeder, Gottfried K; Whitman, Christian P


    Cg10062 is a cis-3-chloroacrylic acid dehalogenase (cis-CaaD) homologue from Corynebacterium glutamicum with an unknown function and an uninformative genomic context. It shares 53% pairwise sequence similarity with cis-CaaD including the six active site amino acids (Pro-1, His-28, Arg-70, Arg-73, Tyr-103, and Glu-114) that are critical for cis-CaaD activity. However, Cg10062 is a poor cis-CaaD: it lacks catalytic efficiency and isomer specificity. Two acetylene compounds (propiolate and 2-butynoate) and an allene compound, 2,3-butadienoate, were investigated as potential substrates. Cg10062 functions as a hydratase/decarboxylase using propiolate as well as the cis-3-chloro- and 3-bromoacrylates, generating mixtures of malonate semialdehyde and acetaldehyde. The two activities occur sequentially at the active site using the initial substrate. With 2,3-butadienoate and 2-butynoate, Cg10062 functions as a hydratase and converts both to acetoacetate. Mutations of the proposed water-activating residues (E114Q, E114D, and Y103F) have a range of consequences from a reduction in wild type activity to a switch of activities (i.e., hydratase into a hydratase/decarboxylase or vice versa). The intermediates for the hydration and decarboxylation products can be trapped as covalent adducts to Pro-1 when NaCNBH3 is incubated with the E114D mutant and 2,3-butadienoate or 2-butynoate, and the Y103F mutant and 2-butynoate. Three mechanisms are presented to explain these findings. One mechanism involves the direct attack of water on the substrate, whereas the other two mechanisms use covalent catalysis in which a covalent bond forms between Pro-1 and the hydration product or the substrate. The strengths and weaknesses of the mechanisms and the implications for Cg10062 function are discussed.

  20. Medical aspects of ketone body metabolism.

    Mitchell, G A; Kassovska-Bratinova, S; Boukaftane, Y; Robert, M F; Wang, S P; Ashmarina, L; Lambert, M; Lapierre, P; Potier, E


    Ketone bodies are produced in the liver, mainly from the oxidation of fatty acids, and are exported to peripheral tissues for use as an energy source. They are particularly important for the brain, which has no other substantial non-glucose-derived energy source. The 2 main ketone bodies are 3-hydroxybutyrate (3HB) and acetoacetate (AcAc). Biochemically, abnormalities of ketone body metabolism can present in 3 fashions: ketosis, hypoketotic hypoglycemia, and abnormalities of the 3HB/AcAc ratio. Normally, the presence of ketosis implies 2 things: that lipid energy metabolism has been activated and that the entire pathway of lipid degradation is intact. In rare patients, ketosis reflects an inability to utilize ketone bodies. Ketosis is normal during fasting, after prolonged exercise, and when a high-fat diet is consumed. During the neonatal period, infancy and pregnancy, times at which lipid energy metabolism is particularly active, ketosis develops readily. Pathologic causes of ketosis include diabetes, ketotic hypoglycemia of childhood, corticosteroid or growth hormone deficiency, intoxication with alcohol or salicylates, and several inborn errors of metabolism. The absence of ketosis in a patient with hypoglycemia is abnormal and suggests the diagnosis of either hyperinsulinism or an inborn error of fat energy metabolism. An abnormal elevation of the 3HB/AcAc ratio usually implies a non-oxidized state of the hepatocyte mitochondrial matrix resulting from hypoxia-ischemia or other causes. We summarize the differential diagnosis of abnormalities of ketone body metabolism, as well as pertinent recent advances in research.

  1. Alterações metabólicas induzidas por isquemia hepática normotérmica experimental e o efeito hepatoprotetor da ciclosporina Induced metabolic alterations due to experimental normothermic hepatic ischemia and the hepatoprotector effect of cyclosporin

    José Huygens Parente Garcia


    hepatic ischemia and the probable hepatoprotective effect cyclosporin. METHOD: Normothermic hepatic ischemia during 60 minutes was induced in the rats. The time-course (0, 1, 6, 24 hours of changes in blood and in the hepatic concentrations of lactate, pyruvate, glucose, ketone bodies and in the ratio of acetoacetate/3-hydroxybutyrate, as well as the cytoplasmic and mitochondrial redox state of the liver cells were determined. A group of animals was daily pre-treated with cyclosporine (10 mg/kg during 4 days until the induction of hepatic ischemia, then they studied 1 hour after hepatic revascularization. Hepatic ischemia caused elevation in the concentrations of lactate in the liver, suggesting that a pronounced level of anaerobic metabolism occurred during the ischemia period. Liver ischemia promoted yet a fall in the concentration and in the ratio of ketone bodies (acetoacetate/3-hydroxybutyrate in the arterial blood in the studied period of one hour post-revascularization, perhaps reflecting impairment of ketogenesis as a result of the ischemic injury. CONCLUSION: The treatment with cyclosporine cause elevation in the concentration of ketone bodies and in the ratio of acetoacetate/3-hydroxybutyrate in the arterial blood 1 hour after reperfusion of the liver, suggesting that these drugs may accelerate the recovery of the ischemic hepatic lesion with reactivation of ketogenesis.

  2. Potential hazards to embryo implantation: A human endometrial in vitro model to identify unwanted antigestagenic actions of chemicals

    Fischer, L.; Deppert, W.R. [Department of Obstetrics and Gynecology, University Hospital Freiburg (Germany); Pfeifer, D. [Department of Hematology and Oncology, University Hospital Freiburg (Germany); Stanzel, S.; Weimer, M. [Department of Biostatistics, German Cancer Research Center, Heidelberg (Germany); Hanjalic-Beck, A.; Stein, A.; Straßer, M.; Zahradnik, H.P. [Department of Obstetrics and Gynecology, University Hospital Freiburg (Germany); Schaefer, W.R., E-mail: [Department of Obstetrics and Gynecology, University Hospital Freiburg (Germany)


    Embryo implantation is a crucial step in human reproduction and depends on the timely development of a receptive endometrium. The human endometrium is unique among adult tissues due to its dynamic alterations during each menstrual cycle. It hosts the implantation process which is governed by progesterone, whereas 17β-estradiol regulates the preceding proliferation of the endometrium. The receptors for both steroids are targets for drugs and endocrine disrupting chemicals. Chemicals with unwanted antigestagenic actions are potentially hazardous to embryo implantation since many pharmaceutical antiprogestins adversely affect endometrial receptivity. This risk can be addressed by human tissue-specific in vitro assays. As working basis we compiled data on chemicals interacting with the PR. In our experimental work, we developed a flexible in vitro model based on human endometrial Ishikawa cells. Effects of antiprogestin compounds on pre-selected target genes were characterized by sigmoidal concentration–response curves obtained by RT-qPCR. The estrogen sulfotransferase (SULT1E1) was identified as the most responsive target gene by microarray analysis. The agonistic effect of progesterone on SULT1E1 mRNA was concentration-dependently antagonized by RU486 (mifepristone) and ZK137316 and, with lower potency, by 4-nonylphenol, bisphenol A and apigenin. The negative control methyl acetoacetate showed no effect. The effects of progesterone and RU486 were confirmed on the protein level by Western blotting. We demonstrated proof of principle that our Ishikawa model is suitable to study quantitatively effects of antiprogestin-like chemicals on endometrial target genes in comparison to pharmaceutical reference compounds. This test is useful for hazard identification and may contribute to reduce animal studies. -- Highlights: ► We compare progesterone receptor-mediated endometrial effects of chemicals and drugs. ► 4-Nonylphenol, bisphenol A and apigenin exert weak

  3. Preparation of Stable Alumina-silica Sol and Its Application in Mullite Fiber%高稳定性氧化铝-二氧化硅溶胶的制备及在莫来石纤维上的应用

    陈根; 张力; 陈立富


    以仲丁醇铝、正硅酸乙酯为原料,制备了铝硅混合溶胶,干法拉丝制备了凝胶纤维,高温热解烧结得到莫来石纤维,研究了溶胶的稳定性以及热处理过程中相演变的过程.研究发现,加入乙酰乙酸乙酯,使其与铝离子螯合,可以有效防止铝溶胶的沉淀,显著改善溶胶的稳定性;聚乙烯吡咯烷酮(PVP)作为成纤助剂显著改善了溶胶的稳定性以及拉丝性能. FTIR、XRD、SEM、TG及DTA分析表明,利用该方法获得的溶胶属于单相溶胶.利用该溶胶制备的凝胶纤维在热处理过程中不形成氧化铝或者二氧化硅结晶相,而是在980℃直接形成莫来石,得到致密、细晶的莫来石纤维.%A stable alumina-silica sol had been prepared by using aluminum sec-butoxide as the source of aluminum and partially hydrolyzed tetraethyl orthosilicate as the source of silicon.FTIR,XRD,SEM,TG and DTA were used to characterize the sol and examine the crystallographic and microstructure evolution of the dry-spun gel fibers during pyrolysis.It is found that ethyl acetoacetate (EAcAc) can effectively retard the precipitation in the sol by chelating with aluminum ions and significantly improve the stability of the sol.Polyvinylpyrrolidone (PVP) is compatible with the sol and has very positive effect on the dry-spinning ability of the spinning sol.No crystalline phases corresponding to alumina or silica have been found during pyrolysis before the formation of crystalline mullite phase at 980℃,indicating that the alumina-silica sol is mono-phasic.When the fiber was fired over 1 000℃,dense mullite fibers were obtained.




    Full Text Available The aim of the study was to analyse the relationship between the body condition and the results of metabolic profile tests done in the milk (DIM of dairy cows in different days. Moreover, critical points in the early pre- and postpartum period were also analysed. In the experiment, blood and urine samples were taken from 1984 clinically healthy cows (from 49 large scale dairy farms in Hungary, selected randomly from various groups of cows with different physiological stage of lactation and gestation, 3-5 hours after the morning feeding. During the experiment body condition scoring (BCS was measured on 1-5 scale, as well. It was concluded, that the BCS (body condition score decreased from the 1st day of lactation (3.48 onwards till the 44th day (2.65 and slightly increased till the day 218 (2.89. The haemoglobin value and the glucose concentration in blood samples were ranging within the physiological range and followed the tendency of BCS and the relationship between them and DIM was (P<0.001. There was a close negative correlation between the NEFA (non-esterified-fatty-acid concentration in blood samples and BCS change and it was found that these values were significantly different (P<0.01 compared to the DIM. The aceto-acetic acid concentration exceeded the upper limit of the physiological range indicating hyperketonaemia at DIM 18. The AST (aspartate aminotransferase, liver-enzyme activity value exceeded the upper limit of physiological range and followed the tendency of BCS change . The urea concentration in the blood exceeded the upper limit of the physiological range in all cows. The NABE (net acid-base empty value in the urine samples indicated acid load in the first two groups of samples (pre-, and post calving. During factor analysis I could differentiate three group factors and one individual. The most important factor is the acid-based factor (with urine pH and NEBA. The results of the present study also confirm that the body

  5. Effects of low-dose recombinant human insulin-like growth factor-I on insulin sensitivity, growth hormone and glucagon levels in young adults with insulin-dependent diabetes mellitus.

    Acerini, C L; Harris, D A; Matyka, K A; Watts, A P; Umpleby, A M; Russell-Jones, D L; Dunger, D B


    Despite recent interest in the therapeutic potential of recombinant human insulin-like growth factor-I (rhIGF-I) in the treatment of diabetes mellitus, its mechanism of action is still not defined. We have studied the effects of low-dose bolus subcutaneous rhIGF-I (40 microg/kg and 20 microg/kg) on insulin sensitivity, growth hormone (GH) and glucagon levels in seven young adults with insulin-dependent diabetes mellitus (IDDM) using a randomized double-blind placebo-controlled crossover study design. Each was subjected to a euglycemic clamp (5 mmol/L) protocol consisting of a variable-rate insulin infusion clamp (6:00 PM to 8:00 AM) followed by a two-dose hyperinsulinemic clamp (insulin infusion of 0.75 mU x kg(-1) x min(-1) from 8 to 10 AM and 1.5 mU x kg(-1) x min(-1) from 10 AM to 12 noon) incorporating [6,6 2H2]glucose tracer for determination of glucose production/utilization rates. Following rhIGF-I administration, the serum IGF-I level (mean +/- SEM) increased (40 microg/kg, 655 +/- 90 ng/mL, P overnight GH level (40 microg/kg, 9.1 +/- 1.4 mU/L, P = .04; 20 microg/kg, 9.6 +/- 2.0 mU/L, P = .12; placebo, 11.3 +/- 1.7 mU/L) and GH pulse amplitude (40 microg/kg, 18.8 +/- 2.9 mU/L, P = .04; 20 microg/kg, 17.0 +/- 3.4 mU/L, P > .05; placebo, 23.0 +/- 3.7 mU/L) were also reduced. No differences in glucagon, IGF binding protein-1 (IGFBP-1), acetoacetate, or beta-hydroxybutyrate levels were found. During the hyperinsulinemic clamp conditions, no differences in glucose utilization were noted, whereas hepatic glucose production was reduced by rhIGF-I 40 microg/kg (P = .05). Our data demonstrate that in subjects with IDDM, low-dose subcutaneous rhIGF-I leads to a dose-dependent reduction in the insulin level for euglycemia overnight that parallels the decrease in overnight GH levels, but glucagon and IGFBP-1 levels remain unchanged. The decreases in hepatic glucose production during the hyperinsulinemic clamp study observed the following day are likely related to GH

  6. Acid Catalysis in Basic Solution: A Supramolecular Host PromotesOrthoformate Hydrolysis

    Pluth, Michael D.; Bergman, Robert G.; Raymond, Kenneth N.


    Though many enzymes can promote chemical reactions by tuning substrate properties purely through the electrostatic environment of a docking cavity, this strategy has proven challenging to mimic in synthetic host-guest systems. Here we report a highly-charged, water soluble, metal-ligand assembly with a hydrophobic interior cavity that thermodynamically stabilizes protonated substrates and consequently catalyzes the normally acidic hydrolysis of orthoformates in basic solution, with rate accelerations of up to 890-fold. The catalysis reaction obeys Michaelis-Menten kinetics, exhibits competitive inhibition, and the substrate scope displays size selectivity consistent with the constrained binding environment of the molecular host. Synthetic chemists have long endeavored to design host molecules capable of selectively binding slow-reacting substrates and catalyzing their chemical reactions. While synthetic catalysts are often site-specific and require certain properties of the substrate to insure catalysis, enzymes are often able to modify basic properties of the bound substrate such as pK{sub a} in order to enhance reactivity. Two common motifs used by nature to activate otherwise unreactive compounds are the precise arrangement of hydrogen-bonding networks and electrostatic interactions between the substrate and adjacent residues of the protein. Precise arrangement of hydrogen bonding networks near the active sites of proteins can lead to well-tuned pK{sub a}-matching, and can result in pK{sub a} shifts of up to eight units, as shown in bacteriorhodopsin. Similarly, purely electrostatic interactions can greatly favor charged states and have been responsible for pK{sub a} shifts of up to five units for acetoacetate decarboxylase. Attempts have been made to isolate the contributions of electrostatic versus covalent interactions to such pK{sub a} shifts; however this remains a difficult challenge experimentally. This challenge emphasizes the importance of synthesizing


    Mahmoud Reza ASHRAFI


    Full Text Available Clinical differential DiagnosisThe organic acidemias are important in the differential diagnosis of metabolic and neurologic derangement in the neonate and of new-onset neurologic signs in the older child.A-Organic aciduriaSeveral disorders, not classified as primary disorders of organic acid metabolism, have a characteristic urinary organic acid profile that suggests the appropriate diagnosis.• Mevalonicaciduria, a disorder of cholesterol biosynthesis, shows mevalonic acid in the urine.• Glutaricacidemia type II, a disorder of fatty acid oxidation, has multiple organic acids in abnormal concentration in urine. These organic acids include ethylmalonic acid, glutaric acid, dicarboxylic acids, and glycine conjugates of medium chain dicarboxylic acids.• The fatty acylCoA-glycine conjugates that signal incomplete fatty acid oxidation and serves as signals to the diagnosis of MCAD defeciency and other disorders of fatty acid oxidation and transport.• Biotinidase deficiency, a disorder of biotin recycling, results in the urinary excretion of several unusual organic acids, including 3-hydroxy-isovaleric, 3-hydroxypropionic, 3-hydroxybutyric acids, and acetoacetate. Propionyl glycine may also be seen.• Mitochondrial diseases with disordered oxidative phosphorylation often demonstrate the presence of abnormal organic acids in the urine.B-AcidosisNon-genetic conditions, such as shock, sepsis, DKA, liver and kidney failure, thiamine deficiency, RTA, some drug intoxication cause acidosis- genetic conditions are include: inherited metabolic disorders of lactate and pyruvate metabolism and oxidative phosphorylation, disorders of the Krebs cycle such as fumarase deficiency.C-HyperammonemiaDisorders of the urea cycle and the hyperammonemia-hypoglycemia syndrome.Neuroimaging• A variety of MRI abnormalities have been described in the organic academia, including distinctive basal ganglia lesions in glutaricacidemia type I (GA I, white matter changes in

  8. Milk metabolome relates enteric methane emission to milk synthesis and energy metabolism pathways.

    Antunes-Fernandes, E C; van Gastelen, S; Dijkstra, J; Hettinga, K A; Vervoort, J


    Methane (CH4) emission of dairy cows contributes significantly to the carbon footprint of the dairy chain; therefore, a better understanding of CH4 formation is urgently needed. The present study explored the milk metabolome by gas chromatography-mass spectrometry (milk volatile metabolites) and nuclear magnetic resonance (milk nonvolatile metabolites) to better understand the biological pathways involved in CH4 emission in dairy cattle. Data were used from a randomized block design experiment with 32 multiparous Holstein-Friesian cows and 4 diets. All diets had a roughage:concentrate ratio of 80:20 (dry matter basis) and the roughage was grass silage (GS), corn silage (CS), or a mixture of both (67% GS, 33% CS; 33% GS, 67% CS). Methane emission was measured in climate respiration chambers and expressed as CH4 yield (per unit of dry matter intake) and CH4 intensity (per unit of fat- and protein-corrected milk; FPCM). No volatile or nonvolatile metabolite was positively related to CH4 yield, and acetone (measured as a volatile and as a nonvolatile metabolite) was negatively related to CH4 yield. The volatile metabolites 1-heptanol-decanol, 3-nonanone, ethanol, and tetrahydrofuran were positively related to CH4 intensity. None of the volatile metabolites was negatively related to CH4 intensity. The nonvolatile metabolites acetoacetate, creatinine, ethanol, formate, methylmalonate, and N-acetylsugar A were positively related to CH4 intensity, and uridine diphosphate (UDP)-hexose B and citrate were negatively related to CH4 intensity. Several volatile and nonvolatile metabolites that were correlated with CH4 intensity also were correlated with FPCM and not significantly related to CH4 intensity anymore when FPCM was included as covariate. This suggests that changes in these milk metabolites may be related to changes in milk yield or metabolic processes involved in milk synthesis. The UDP-hexose B was correlated with FPCM, whereas citrate was not. Both metabolites were

  9. Exogenous ketone supplements reduce anxiety-related behavior in Sprague-Dawley and Wistar Albino Glaxo/Rijswijk rats

    Csilla Ari


    Full Text Available Nutritional ketosis has been proven effective for seizure disorders and other neurological disorders. The focus of this study was to determine the effects of ketone supplementation on anxiety-related behavior in Sprague-Dawley (SPD and Wistar Albino Glaxo/Rijswijk (WAG/Rij rats. We tested exogenous ketone supplements added to food and fed chronically for 83 days in SPD rats and administered sub-chronically for 7 days in both rat models by daily intragastric gavage bolus followed by assessment of anxiety measures on elevated plus maze (EPM. The groups included standard diet (SD or SD + ketone supplementation. Low-dose ketone ester (LKE (1,3-butanediol-acetoacetate diester, ~10 g/kg/day, LKE, high dose ketone ester (HKE (~25 g/kg/day, HKE, beta-hydroxybutyrate-mineral salt (βHB-S (~25 g/kg/day, KS, and βHB-S + medium chain triglyceride (MCT (~25 g/kg/day, KSMCT were used as ketone supplementation for chronic administration. To extend our results, exogenous ketone supplements were also tested sub-chronically on SPD rats (KE, KS and KSMCT; 5 g/kg/day and on WAG/Rij rats (KE, KS and KSMCT; 2.5 g/kg/day. At the end of treatments behavioral data collection was conducted manually by a blinded observer and with a video-tracking system, after which blood βHB and glucose levels were measured. Ketone supplementation reduced anxiety on EPM as measured by less entries to closed arms (sub-chronic KE and KS: SPD rats and KSMCT: WAG/Rij rats, more time spent in open arms (sub-chronic KE: SPD and KSMCT: WAG/Rij rats; chronic KSMCT: SPD rats, more distance travelled in open arms (chronic KS and KSMCT: SPD rats, and by delayed latency to entrance to closed arms (chronic KSMCT: SPD rats, when compared to control. Our data indicates that chronic and sub-chronic ketone supplementation not only elevated blood βHB levels in both animal models, but reduced anxiety-related behavior. We conclude that ketone supplementation may represent a promising anxiolytic strategy

  10. Serum metabolomics studies of abnormal savda syndrome rat model%基于异常黑胆质证载体大鼠模型血清代谢组学研究

    阿衣古丽·玉努斯; 哈木拉提·吾甫尔; 帕丽旦·麦麦提; 阿布力孜·阿卜杜扎依尔; 甫拉提·吐尔逊; 库热西·玉努斯


    Objective:To investigate metabolites content variation in occurrence and development of abnormal savda syndrome.Methods:Based on establishing abnormal savda syndrome rat model,based metabolomic analysis was performed on serum of abnormal savda syndrome rat model groups and control groups to identify differential metabolites.Results:Compared with normal control groups,there was significant serum content difference of VLDL,LDL,unsaturated fatty acid,acetoacetic acid,acetone,phenylalanine,creatine,inositol in model groups (P<0.05).Conclusion:The serum metabolic spectrum in the model groups differed from those in the control groups.The occurrence and development of abonormal savda syndrome can cause metabolic disturbance of creatine,carbohydrate,fat and amino acid.%目的:探讨异常黑胆质型体液在异常黑胆质证发生、发展过程中对大鼠血清代谢组分的影响.方法:在建立异常黑胆质证大鼠模型的基础上,运用代谢组学研究技术,对异常黑胆质证及正常大鼠血清进行代谢组学研究,分析两组大鼠血清差异代谢组分.结果:异常黑胆质证组与正常对照组大鼠血清间存在差异代谢组分,与正常对照组相比,异常黑胆质证组大鼠血清中VLDL、LDL、不饱和脂肪酸、乙酰乙酸、丙酮等代谢组分的含量升高,苯丙氨酸、肌酸、肌醇等代谢组分的含量下降(P<0.05).结论:异常黑胆质证组与正常对照组大鼠血清代谢物谱存在差异,异常黑胆质证大鼠存在糖、脂肪、蛋白质及磷酸肌酸代谢紊乱.

  11. Synthesis, Antibacterial and Antifungal Activity of Some New Pyrazoline and Pyrazole Derivatives

    Seham Y. Hassan


    Full Text Available A series of 2-pyrazolines 5–9 have been synthesized from α,β-unsaturated ketones 2–4. New 2-pyrazoline derivatives 13–15 bearing benzenesulfonamide moieties were then synthesized by condensing the appropriate chalcones 2–4 with 4-hydrazinyl benzenesulfonamide hydrochloride. Ethyl [1,2,4] triazolo[3,4-c][1,2,4]triazino[5,6-b]-5H-indole-5-ethanoate (26 and 1-(5H-[1,2,4]triazino[5,6-b] indol-3-yl-3-methyl-1H-pyrazol-5(4H-one (32 were synthesized from 3-hydrazinyl-5H-[1,2,4]triazino[5,6-b]indole (24. On the other hand ethyl[1,2,4]triazolo[3,4-c][1,2,4]triazino[5,6-b]-5,10-dihydroquinoxaline- 5-ethanoate (27 and 1-(5,10-dihydro-[1,2,4]triazino[5,6-b]quinoxalin-3-yl-3-methyl-1H-pyrazol-5(4H-one (33 were synthesized from 3-hydrazinyl-5,10-dihydro-[1,2,4]triazino[5,6-b]quinoxaline (25 by reaction with diethyl malonate or ethyl acetoacetate, respectively. Condensation of 6,6-dimethyl-4-oxo-4,5,6,7-tetrahydro-1H-indole-2-carbaldehyde (1' with compound 24 or 25 afforded the corresponding Schiff's bases 36 and 37, respectively. Reaction of the Schiff's base 37 with benzoyl hydrazine or acetic anhydride afforded benzohydrazide derivative 39 and the cyclized compound 40, respectively. Furthermore, the pyrazole derivatives 42–44 were synthesized by cyclization of hydrazine derivative 25 with the prepared chalcones 2–4. All the newly synthesized compounds have been characterized on the basis of IR and 1H-NMR spectral data as well as physical data. Antimicrobial activity against the organisms E. coli ATCC8739 and P. aeruginosa ATCC 9027 as examples of Gram-negative bacteria, S. aureus ATCC 6583P as an example of Gram-positive bacteria and C. albicans ATCC 2091 as an example of a yeast-like fungus have been studied using the Nutrient Agar (NA and Sabouraud Dextrose Agar (SDA diffusion methods. The best performance was found for the compounds 16, 17, 19 and 20.

  12. C-Myc induced compensated cardiac hypertrophy increases free fatty acid utilization for the citric acid cycle.

    Olson, Aaron K; Ledee, Dolena; Iwamoto, Kate; Kajimoto, Masaki; O'Kelly Priddy, Colleen; Isern, Nancy; Portman, Michael A


    The protooncogene C-Myc (Myc) regulates cardiac hypertrophy. Myc promotes compensated cardiac function, suggesting that the operative mechanisms differ from those leading to heart failure. Myc regulation of substrate metabolism is a reasonable target, as Myc alters metabolism in other tissues. We hypothesize that Myc induced shifts in substrate utilization signal and promote compensated hypertrophy. We used cardiac specific Myc-inducible C57/BL6 male mice between 4-6 months old that develop hypertrophy with tamoxifen (tam) injections. Isolated working hearts and (13)Carbon ((13)C)-NMR were used to measure function and fractional contributions (Fc) to the citric acid cycle by using perfusate containing (13)C-labeled free fatty acids, acetoacetate, lactate, unlabeled glucose and insulin. Studies were performed at pre-hypertrophy (3-days tam, 3dMyc), established hypertrophy (7-days tam, 7dMyc) or vehicle control (Cont). Non-transgenic siblings (NTG) received 7-days tam or vehicle to assess drug effect. Hypertrophy was assessed by echocardiograms and heart weights. Western blots were performed on key metabolic enzymes. Hypertrophy occurred in 7dMyc only. Cardiac function did not differ between groups. Tam alone did not affect substrate contributions in NTG. Substrate utilization was not significantly altered in 3dMyc versus Cont. The free fatty acid FC was significantly greater in 7dMyc versus Cont with decreased unlabeled Fc, which is predominately exogenous glucose. Free fatty acid flux to the citric acid cycle increased while lactate flux was diminished in 7dMyc compared to Cont. Total protein levels of a panel of key metabolic enzymes were unchanged; however total protein O-GlcNAcylation was increased in 7dMyc. Substrate utilization changes for the citric acid cycle did not precede hypertrophy; therefore they are not the primary signal for cardiac growth in this model. Free fatty acid utilization and oxidation increase at established hypertrophy. Understanding the

  13. NADPH-dependent reductive biotransformation with Escherichia coli and its pfkA deletion mutant: influence on global gene expression and role of oxygen supply.

    Siedler, Solvej; Bringer, Stephanie; Polen, Tino; Bott, Michael


    An Escherichia coli ΔpfkA mutant lacking the major phosphofructokinase possesses a partially cyclized pentose phosphate pathway leading to an increased NADPH per glucose ratio. This effect decreases the amount of glucose required for NADPH regeneration in reductive biotransformations, such as the conversion of methyl acetoacetate (MAA) to (R)-methyl 3-hydroxybutyrate (MHB) by an alcohol dehydrogenase from Lactobacillus brevis. Here, global transcriptional analyses were performed to study regulatory responses during reductive biotransformation. DNA microarray analysis revealed amongst other things increased expression of soxS, supporting previous results indicating that a high NADPH demand contributes to the activation of SoxR, the transcriptional activator of soxS. Furthermore, several target genes of the ArcAB two-component system showed a lower mRNA level in the reference strain than in the ΔpfkA mutant, pointing to an increased QH2 /Q ratio in the reference strain. This prompted us to analyze yields and productivities of MAA reduction to MHB under different oxygen regimes in a bioreactor. Under anaerobic conditions, the specific MHB production rates of both strains were comparable (7.4 ± 0.2 mmolMHB  h(-1)  gcdw (-1) ) and lower than under conditions of 15% dissolved oxygen, where those of the reference strain (12.8 mmol h(-1)  gcdw (-1) ) and of the ΔpfkA mutant (11.0 mmol h(-1)  gcdw (-1) ) were 73% and 49% higher. While the oxygen transfer rate (OTR) of the reference strain increased after the addition of MAA, presumably due to the oxidation of the acetate accumulated before MAA addition, the OTR of the ΔpfkA strain strongly decreased, indicating a very low respiration rate despite sufficient oxygen supply. The latter effect can likely be attributed to a restricted conversion of NADPH into NADH via the soluble transhydrogenase SthA, as the enzyme is outcompeted in the presence of MAA by the recombinant NADPH-dependent alcohol

  14. Postprandial metabolomics: A pilot mass spectrometry and NMR study of the human plasma metabolome in response to a challenge meal

    Karimpour, Masoumeh; Surowiec, Izabella; Wu, Junfang [Computational Life Science Cluster (CLiC), Department of Chemistry, Umeå University, 90187 Umeå (Sweden); Gouveia-Figueira, Sandra [Computational Life Science Cluster (CLiC), Department of Chemistry, Umeå University, 90187 Umeå (Sweden); Department of Pharmacology and Clinical Neuroscience, Umeå University, Umeå (Sweden); Pinto, Rui [Computational Life Science Cluster (CLiC), Department of Chemistry, Umeå University, 90187 Umeå (Sweden); Bioinformatics Infrastructure for Life Sciences (Sweden); Trygg, Johan [Computational Life Science Cluster (CLiC), Department of Chemistry, Umeå University, 90187 Umeå (Sweden); Zivkovic, Angela M. [Department of Nutrition, University of California, Davis, One Shields Ave, CA 95616 (United States); Nording, Malin L., E-mail: [Computational Life Science Cluster (CLiC), Department of Chemistry, Umeå University, 90187 Umeå (Sweden)


    The study of postprandial metabolism is relevant for understanding metabolic diseases and characterizing personal responses to diet. We combined three analytical platforms – gas chromatography-mass spectrometry (GC-MS), liquid chromatography-mass spectrometry (LC-MS) and nuclear magnetic resonance (NMR) – to validate a multi-platform approach for characterizing individual variation in the postprandial state. We analyzed the postprandial plasma metabolome by introducing, at three occasions, meal challenges on a usual diet, and 1.5 years later, on a modified background diet. The postprandial response was stable over time and largely independent of the background diet as revealed by all three analytical platforms. Coverage of the metabolome between NMR and GC-MS included more polar metabolites detectable only by NMR and more hydrophobic compounds detected by GC-MS. The variability across three separate testing occasions among the identified metabolites was in the range of 1.1–86% for GC-MS and 0.9–42% for NMR in the fasting state at baseline. For the LC-MS analysis, the coefficients of variation of the detected compounds in the fasting state at baseline were in the range of 2–97% for the positive and 4–69% for the negative mode. Multivariate analysis (MVA) of metabolites detected with GC-MS revealed that for both background diets, levels of postprandial amino acids and sugars increased whereas those of fatty acids decreased at 0.5 h after the meal was consumed, reflecting the expected response to the challenge meal. MVA of NMR data revealed increasing postprandial levels of amino acids and other organic acids together with decreasing levels of acetoacetate and 3-hydroxybutanoic acid, also independent of the background diet. Together these data show that the postprandial response to the same challenge meal was stable even though it was tested 1.5 years apart, and that it was largely independent of background diet. This work demonstrates the efficacy of a

  15. 七叶鬼灯檠挥发油及不同提取物抑菌活性的研究%Study on antimicrobial activities of the essential oil and various extracts of Rodgersia aesculifolia Batalin

    王健; 黄慧; 王喆之; 张媛


    研究了七叶鬼灯檠根状茎的挥发油及不同提取物的体外抑菌活性,为七叶鬼灯檠资源开发利用提供科学依据.采用滤纸片法和牛津杯法对七叶鬼灯檠根状茎挥发油及其甲醇、丙酮和乙酸乙酯提取物体外抑菌能力进行评价.结果表明,七叶鬼灯檠根状茎挥发油表现出较强的抑菌活性,对肺炎克雷伯氏菌抑制作用最强,其次为巨大芽孢杆菌和粘红酵母,抑菌圈直径分别为(29.11±0.31)、(26.77±0.35)、(26.20±0.62)mm.在不同溶剂提取物中,丙酮提取物和甲醇提取物均有明显的抑菌效果,解脂假丝酵母对提取物最敏感.七叶鬼灯檠的挥发油及提取物含有天然抑菌活性物质,具有光谱的抑菌效果,值得进一步开发利用.%Examined the antimicrobial activities of the essential oil and various extracts of Rodgersia aesculifolia Batalin.Disc diffusion method and agar well-diffusion method were used to evaluate the in vitro antimicrobial activities of the essential oil and various extracts( methanol extract,acetone extract and acetoacetate extract)of R. aesculifolia , respectively. The results indicated that the essential oil showed strong antimicrobial activity against Klebsiella pneumoniae subsp.Pneumoniae(with(29.11±0.31) mm inhibition zone diameter).The oil was found to possess high antimicrobial activities against Bacillus megaterium(26.77±0.35) mm and Rhodotorula glatinis(26.20 ±0.62) mm. Among the different extracts, acetone and methanol extracts exhibited significant antimicrobial activities against all tested microbial species,especially against Candida lipoiytica.The essential oil and extracts of Raesculifoiia has potential utilities as a natural antibacterial substance, and it is of worth for further study.

  16. Scientific Opinion on the safety and efficacy of primary aliphatic saturated or unsaturated alcohols/aldehydes/acids/acetals/esters with a second primary, secondary or tertiary oxygenated functional group including aliphatic lactones (chemical group 9 when used as flavourings for all animal species

    EFSA Panel on Additives and Products or Substances used in Animal Feed (FEEDAP


    Full Text Available

    Chemical group 9 consists of primary aliphatic saturated or unsaturated alcohols/aldehydes/acids/acetals/esters with a second primary, secondary or tertiary oxygenated functional group including aliphatic lactones, of which 30 are currently authorised for use as flavours in food. The FEEDAP Panel was unable to perform an assessment of 2-oxopropanal because of issues related to the purity of the compound. The FEEDAP Panel concludes that lactic acid, succinic acid, fumaric acid, 4-oxovaleric acid, ethyl lactate, butyl lactate, butyl-O-butyryllactate, hex-3-enyl lactate, hexyl lactate, ethyl acetoacetate, ethyl 4-oxovalerate, diethylsuccinate and diethyl malonate are considered to be safe for all animal species at the use levels proposed when used as feed flavourings; octano-1,4-lactone, nonano-1,4-lactone, decano-1,4-lactone and undecano-1,4-lactone are safe at 20 mg/kg complete feed; butyro-1,4-lactone, pentano-1,4-lactone, hexano-1,4-lactone, heptano-1,4-lactone, octano-1,5-lactone, nonano-1,5-lactone, decano-1,5-lactone and undecano-1,5-lactone at 5 mg/kg complete feed; dodecano-1,4-lactone, dodecano-1,5-lactone, tetradecano-1,5-lactone, and pentadecano-1,15-lactone at a maximum of 1.5 mg/kg complete feed for cattle, salmonids and non food producing animals and of 1 mg/kg complete feed for pigs and poultry. No safety concern was identified for the consumer from the use of compounds belonging to CG 9 up to the highest safe level in feedingstuffs for all animal species. All compounds should be considered as irritants to skin, eyes and respiratory tract, and as skin sensitizers. The compounds do not pose a risk to the environment when used at concentrations considered safe for the target species. Since all compounds are used in food as flavourings, no further demonstration of efficacy is necessary.

  17. Changes in energetic metabolism of Biomphalaria glabrata (Mollusca, Planorbidae in response to exogenous calcium

    L. D. Silva

    Full Text Available Abstract Calcium is considered an essential element for the metabolism of aquatic snail Biomphalaria glabrata (Say, 1818, intermediate host of Schistosoma mansoni Sambon, 1907 in Brazil, and represents a limiting factor to its distribution and adaptation to the environment. This study investigated the effect of different concentrations of exogenous CaCO3 on the energetic metabolism of B. glabrata for better understanding the physiological interference of chemical elements dissolved in the environment with the physiology of this species. Sixty-day-old snails were distributed into six groups, five exposed to different concentrations of CaCO3 (20, 40, 60, 80 and 100 mg/L and a control group. The exposure to CaCO3 was assessed over time, with analysis of 15 snails of each group in the following intervals: 1, 14, 21 or 30 days for hemolymph extraction. Concentrations of calcium and glucose in the hemolymph were determined by commercial kits, and organic acids were extracted using an ion exchange column and analyzed by high-performance liquid chromatography. Concentration of calcium in the hemolymph showed no significant difference (p>0.05 from the control group and between the concentrations tested. Concentration of glucose decreased (p<0.05 in the treatments of exposure to 20 and 40 mg/L and increased when exposed to 80 and 100 mg/L CaCO3 compared to control and to other concentrations tested over 30 days. The organic acids pyruvate, oxaloacetate, citrate, succinate, fumarate, beta-hydroxybutyrate and lactate presented increased concentrations, while propionate and acetoacetate, decreased concentrations, when exposed to CaCO3 compared to control. Considering the influence of different periods of exposure to CaCO3, on the 14th day, there were stronger alterations in the metabolism of B. glabrata. In conclusion, exposure to CaCO3 reduced the concentration of glucose, which is metabolized into pyruvate, the final product of glycolysis, and also

  18. Diurnal variation of blood ketone bodies in insulin-dependent diabetes mellitus and noninsulin-dependent diabetes mellitus patients: the relationship to serum C-peptide immunoreactivity and free insulin.

    Ubukata, E


    We examined whether the rise in ketone body concentration around midnight and in the early morning was due to the lack of free insulin (IRI) or excess of insulin counterregulatory hormones such as human growth hormone (hGH), cortisol and glucagon in noninsulin-dependent diabetes mellitus (NIDDM) and insulin-dependent diabetes mellitus (IDDM) patients and whether the monitoring of blood ketone body concentration was clinically useful as an index of metabolic control for deciding to increase or decrease the insulin dose in the treatment of diabetes mellitus. Serum levels of 3-hydroxybutyrate (3-OHBA), acetoacetate (AcAc) and 3-OHBA/AcAc ratio before breakfast were significantly increased in insulin-treated NIDDM patients with well-controlled fasting plasma glucose levels and IDDM patients compared to those in normal subjects. Mirror image diurnal changes were found between serum concentrations of 3-OHBA and serum C-peptide or free IRI in normal subjects and NIDDM patients treated with diet alone or sulfonylurea during the 24-hour daily profiles. However, there were no correlations between 3-OHBA and free IRI in the NIDDM patients treated with insulin and IDDM patients who had a much larger increase in the mean concentration of serum 3-OHBA at 6 a.m. caused by a low concentration of free IRI. Counterregulatory hormones were not increased in IDDM patients compared to normal subjects in the early morning. Cortisol/free IRI and hGH/free IRI molar ratios were significantly increased in NIDDM and IDDM patients compared to normal subjects in the early morning, but glucagon/free IRI molar ratio was not changed between IDDM and normal subjects. In conclusion, the early morning rising of ketone body concentration in insulin-treated diabetic patients, particularly IDDM patients, is due to the absolute lack of free IRI and/or the relative lack of free IRI to the levels of hGH or cortisol, and the monitoring of 3-OHBA is clinically useful as a more sensitive index of metabolic

  19. C-Myc Induced Compensated Cardiac Hypertrophy Increases Free Fatty Acid Utilization for the Citric Acid Cycle

    Olson, Aaron; Ledee, Dolena; Iwamoto, Kate; Kajimoto, Masaki; O' Kelly-Priddy, Colleen M.; Isern, Nancy G.; Portman, Michael A.


    The protooncogene C-Myc (Myc) regulates cardiac hypertrophy. Myc promotes compensated cardiac function, suggesting that the operative mechanisms differ from those leading to heart failure. Myc regulation of substrate metabolism is a reasonable target, as Myc alters metabolism in other tissues. We hypothesize that Myc-induced shifts in substrate utilization signal and promote compensated hypertrophy. We used cardiac specific Myc-inducible C57/BL6 male mice between 4-6 months old that develop hypertrophy with tamoxifen (tam). Isolated working hearts and 13Carbon (13C )-NMR were used to measure function and fractional contributions (Fc) to the citric acid cycle by using perfusate containing 13C-labeled free fatty acids, acetoacetate, lactate, unlabeled glucose and insulin. Studies were performed at pre-hypertrophy (3-days tam, 3dMyc), established hypertrophy (7-days tam, 7dMyc) or vehicle control (cont). Non-transgenic siblings (NTG) received 7-days tam or vehicle to assess drug effect. Hypertrophy was confirmed by echocardiograms and heart weights. Western blots were performed on key metabolic enzymes. Hypertrophy occurred in 7dMyc only. Cardiac function did not differ between groups. Tam alone did not affect substrate contribution in NTG. Substrate utilization was not significantly altered in 3dMyc versus cont. The free fatty acid FC was significantly greater in 7dMyc vs cont with decreased unlabeled Fc, which is predominately exogenous glucose. Free fatty acid flux to the citric acid cycle increased while lactate flux was diminished in 7dMyc compared to cont. Total protein levels of a panel of key metabolic enzymes were unchanged; however total protein O-GlcNAcylation was increased in 7dMyc. Substrate utilization changes did not precede hypertrophy; therefore they are not the primary signal for cardiac growth in this model. Free fatty acid utilization and oxidation increase at established hypertrophy. Understanding the mechanisms whereby this change maintained

  20. 1H nuclear magnetic resonance spectroscopy-basedmetabonomic study in patients with cirrhosis and hepaticencephalopathy


    AIM To identify plasma metabolites used as biomarkersin order to distinguish cirrhotics from controls and encephalopathics.METHODS: A clinical study involving stable cirrhoticpatients with and without overt hepatic encephalopathywas designed. A control group of healthy volunteers wasused. Plasma from those patients was analysed using1H - nuclear magnetic resonance spectroscopy. Weused the Carr Purcell Meiboom Gill sequence to processthe sample spectra at ambient probe temperature. Weused a gated secondary irradiation field for water signalsuppression. Samples were calibrated and referencedusing the sodium trimethyl silyl propionate peak at0.00 ppm. For each sample 128 transients (FID's)were acquired into 32 K complex data points over aspectral width of 6 KHz. 30 degree pulses were appliedwith an acquisition time of 4.0 s in order to achievebetter resolution, followed by a recovery delay of 12s, to allow for complete relaxation and recovery ofthe magnetisation. A metabolic profile was created forstable cirrhotic patients without signs of overt hepaticencephalopathy and encephalopathic patients as wellas healthy controls. Stepwise discriminant analysis wasthen used and discriminant factors were created todifferentiate between the three groups.RESULTS: Eighteen stabled cirrhotic patients, eighteenpatients with overt hepatic encephalopathy and seventeenhealthy volunteers were recruited. Patients with cirrhosishad significantly impaired ketone body metabolism, ureasynthesis and gluconeogenesis. This was demonstratedby higher concentrations of acetoacetate (0.23 ± 0.02vs 0.05 ± 0.00, P 〈 0.01), and b-hydroxybutarate (0.58± 0.14 vs 0.08 ± 0.00, P 〈 0.01), lower concentrationsof glutamine (0.44 ± 0.08 vs 0.63 ± 0.03, P 〈 0.05),histidine (0.16 ± 0.01 vs 0.36 ± 0.04, P 〈 0.01) andarginine (0.08 ± 0.01 vs 0.14 ± 0.02, P 〈 0.03) andhigher concentrations of glutamate (1.36 ± 0.25 vs0.58 ± 0.04, P 〈 0.01), lactate (1

  1. Lipoperoxidação, perda de viabilidade celular e diminuição da liberação de IL-8 de neutrófilos humanos na presença de corpos cetônicos



    Full Text Available

    Portadores de diabetes tipo-1 são acometidos por episódios freqüentes de acidose causada pelo aumento no metabolismo de ácido graxos com conseqüente acúmulo de acetoacetato (AcAc e β-hidroxibutirato (β -OB. Neste trabalho estudou-se o efeito de concentrações patológicas destes metabólitos na lipoperoxidação, viabilidade e liberação da quimiocina CXCL8 (IL-8 por neutrófilos em cultura. Neutrófilos de indivíduos saudáveis foram isolados por gradiente de densidade (Histopaque 1077/1119 e incubados com os corpos cetônicos. A lipoperoxidação foi determinada pela presença de substâncias reagentes ao ácido tiobarbitúrico (TBARS. A viabilidade celular foi avaliada pela liberação da enzima lactato desidrogenase. A liberação de CXCL8 para o meio extracelular foi medida após cultura de 24 horas de neutrófilos estimulados por zymosan opsonizado por ensaio imunoenzimático (ELISA. O AcAc causou um aumento na lipoperoxidação dos neutrófilos e este efeito foi dependente da sua concentração (p < 0.05; r = 0.99146; não se observou efeito do β-HOB. No estudo do efeito citotóxico, houve aumento dose-dependente da liberação da LDH até 40 mM de AcAc (p < 0.05; não se observou efeito do β-HOB. A liberação de CXCL8 foi suprimida de modo dose dependente por AcAc e β-HOB. Estes resultados sugerem que o acúmulo de corpos cetônicos pode contribuir para aumentar o tempo de remissão de doenças e mesmo estar relacionado com a gravidade destas em indivíduos diabéticos. Palavras-chave: Acetoacetato. Lipoperoxidação. Neutrófilos. Interleucina-8. Diabetes mellitus. ABSTRACT Lipoperoxidation, loss of cell viability and inhibition of release of IL-8 BY human neutrophils in the presence of ketone bodies Type-1 diabetes patients suffer from frequent episodes of acidosis caused by an increased fatty acid metabolism and consequently increased plasma level of acetoacetate (AcAc and β-hydroxybutyrate (

  2. Plasma Metabonomics Analysis of Tumor Patients of Phlegm-Stasis Syndrome%痰瘀证肿瘤患者血浆代谢组学研究

    巴吐尔·买买提明; 哈木拉提·吾甫尔


    Objective To study metabonomic changes in plasma of tumor patients of phlegm-stasis syndrome by Chinese medicine and their in vivo metabolic mechanism. Methods 1H nuclear magnetic resonance (NMR) based metabonomic analysis was performed on plasma samples from 356 tumor patients of the phlegmstasis syndrome and 104 tumor patients of the non-phlegm-stasis syndrome, and 50 healthy subjects. The spectrogram integral results were analyzed by orthogonal partial least-squares discriminant analysis (OPLS-DA). Results Compared with healthy subjects, various amino acids including leucine, alanine, citrulline, tyrosine, histidine, arginine, methionine, isoleucine, valine, acetylcysteine, etc. in the plasma of patients of the phlegm-stasis syndrome were significantly lowered (P <0. 05). Glucose, glycoprotein, glutamine, myo-inositol, lactic acid, choline, creatine also significantly decreased ( P < 0. 05). But the plasmal formic acid, acetone, acetic acid, acetoacetate, pyruvate, β-hydrocxy butyrate, carnitine, malonic acid, and unsaturated fatty acid, very low density lipoprotein cholesterol (VLDL-C), low density lipoprotein cholesterol (LDL-C) increased in tumor patients of the phlegm-stasis syndrome. Compared with tumor patients of non-phlegm-stasis syndrome, patients of the phlegm-stasis syndrome had obvious lower plasmal contents of leucine, alanine, citrulline, tyrosine, histidine, isoleucine, valine, glutamine, myo-inositol, scyllo-inositol, lactic acid, creatine ( P < 0. 05), higher plasmal contents of acetone, acetoacetate, unsaturated fatty acid, VLDL-C, α-glucose, β-glucose, glycoprotein, and so on ( P<0. 05). Conclusions Tumor patients of the phlegm-stasis syndrome had strengthened in vivo fat metabolism and lowered various amino acids. The decreased antioxidation capacities resulted in aggravated cell membrane injuries. The in vivo metabolic disorder was more severe in tumor patients of the phlegm-stasis syndrome than in tumor patients of the non

  3. Effects of extracts of Dragon's blood on fibroblast proliferation and extracellular matrix hyaluronic acid%血竭提取物对成纤维细胞增殖及合成透明质酸的影响

    李丹; 惠瑞; 胡咏武; 韩岩; 郭树忠


    Objective To investigate the effects of Dragon' s blood extract on proliferation and secret extracellular matrix function of fibroblasts in vitro.Methods Dragon ' s blood was extracted by chloroform,acetoacetic ester,alcohol.Human fibroblast were cultured in vitro in media containing gradient dilutions of Dragon's blood extracts (0.002,0.02,0.2,2,20 mg/ml),which was followed by cell proliferation assessed with MTT assay on 0,12,24,36,48,60,72 h.Under the optimal concentration,the cell growth curves were drawn and the flow cytometry (FCM) was used to determine the changes of cell cycle.On 0,12,24,36,48,60,72 h,the concentration of hyaluronic acid in the supernatant of fibroblast culture was measured by radioimmunoassay.Results 0.2-2 mg/ml Dragon' s blood extracts enhanced the proliferation of fibroblasts in a dose-dependent manner.2 mg/ml was the optimal dilution of Dragon' s blood extract,and it increased the ratio of S cells in cell cycle[(25.80 ± 3.10) %] than control group[(7.50 ± 0.70)%,P < 0.01].From 12 h to 72 h,in 2 mg/ml Dragon's blood group,concentration of Hyaluronic acid secreted by fibroblasts gradually increased,but were less than control(P < 0.01).Conclusions Dragon ' s blood acetoacetic ester extract improved the proliferation of cultured human fibroblasts in vitro,might be beneficial to promote wound healing.%目的 研究血竭提取物对成纤维细胞生物学作用的影响.方法 将血竭经过氯仿、乙酸乙酯、乙醇依次回流提取分别得到3种血竭提取液,培养液稀释后行成纤维细胞培养.噻唑蓝法MTT检测浓度分别为0.002、0.02、0.2、2、20 mg/ml的血竭提取物,在0、12、24、36、48、60、72h7个检测点对体外培养成纤维细胞增殖的影响,并绘制最适浓度下细胞生长曲线.流式细胞术FCM分析最适浓度培养下成纤维细胞的细胞周期变化.应用放射免疫分析法检测0、12、24、36、48、60、72 h细胞培养上清液中透明质酸(hyaluronic acid

  4. Detection and Validated Quantification of 11 Sedative-Hypnotics in Human Plasma by UPLC-MS/MS Method%超高效液相色谱-串联质谱法同时检测人血浆中11种镇静催眠药浓度∗

    安静; 董占军; 魏欣; 白万军; 宋浩静


    Objective To develop a rapid ultra high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/ MS) for determination of 11 sedative-hypnotics in human plasma. Methods The plasma samples were extracted with dichloromethane:n-hexane:acetoacetate (5∶4∶1). The separation was performed on a Waters ACQUITY UPLC HSS T3 column (2.1 mm×100 mm,1.8 μm) using the mobile phase composed of acetonitrile-0.1% ammonium solution at a flow rate of 0.2 mL•min-1 in gradient elution mode. The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring (MRM) via electrospray ionization (ESI) source in positive mode. Results Good linear relation was obtained over the investigated concentration range, with all correlation coefficients higher than 0.99. The limit of detection was 200 pg•mL-1 for dexzopiclone,and 10- 20 pg•mL-1 for other sedative-hypnotics. The intra- and inter-day RSD of 11 sedative-hypnotics were no more than 15.0%. The recoveries were in the range of 72.3%-108.0%, and the matrix effects were approximately 0.86- 1. 12. Conclusion The method is rapid, sensitive and reliable, and suitable for the simultaneous determination of 11 sedative-hypnotics in human plasma.%目的:建立超高效液相色谱-串联质谱(UPLC-MS/ MS)测定人血浆中11种镇静催眠药的浓度。方法血样采用二氯甲烷:正己烷:乙酸乙酯(5:4:1)进行萃取,Waters ACQUITY UPLC HSS T3(2.1 mm×100 mm,1.8μm)色谱柱进行分离,乙腈-0.1%氨溶液为流动相进行梯度洗脱,流速0.2 mL•min-1,通过电喷雾离子源(ESI),多重反应监测(MRM)正离子模式进行检测。结果11种镇静催眠药在所测质量浓度范围内呈现良好的线性关系,相关系数均>0.99,除右佐匹克隆检测限为200 pg•mL-1,其他物质检测限为10~20 pg•mL-1,11种被分析物的日内、日间 RSD 均<15.0%,提取回收率范围为72.3%~108.0%,基质效应范围为0

  5. Determination of fluorouracil concentration in tissues by HPLC%高效液相色谱法测定氟尿嘧啶的组织浓度

    余俊先; 蔡军; 史丽敏; 王康里; 罗晓; 温爱萍; 李任; 王汝龙


    Objective: An HPLC method was developed to determine the concentration of fluorouracil in tissues, which was a metabolite of capecitabine in tissues. Methods: Plasma contained fluorouracil with bromouracil as the internal standard was extracted with acetoacetate, then the organic layer was evaporated to dryness under nitrogen stream and the residue was reconstituted with mobile phase for HPLC assay. Gastric carcinoma tissues and perienchyma through exairesis were homogenated with blank plasma and dealt with the same as described above. The mobile phase consisted of acetointrile: water (1: 99) at a flow rate of 1.0 mL·min-1, and the detection wavelength was 254 nm. Results: The retention times of fluorouracil and bromouracil were 3.20 minutes and 7.50 minutes, respectively. The assay exhibited excellent linearity with the range of 0.02 - 1.0 μg·mL-1 (r = 0.999 6). The reproducibility, extraction recovery, the variability of intra- and inter- day assay were consistent with requirement for determining biological specimen. Fluorouracil, metabolite of capecitabine, in gastric carcinoma central tissues (1.095 μg·g-1) was significantly higher than that in normal gastric tissues (0.047 μg·g-1). Conclusion: The method is suitable for determining the concentration of fluorouracil in tissues, and applicable to the study of capecitabine metabolism in human.%目的:建立高效液相色谱法测定卡培他滨代谢物氟尿嘧啶的组织浓度.方法:取氟尿嘧啶血浆溶液,加入溴尿嘧啶为内标,经乙酸乙酯提取后氮气吹干,流动相复溶进样分析;手术切除的胃癌及其周边组织用空白血浆匀浆,匀浆液处理方法同上.流动相为乙腈-水(1:99),流速1.0mL·min-1,检测波长254nm.结果:氟尿嘧啶与内标溴尿嘧啶保留时间分别为3.20min和7.50min.氟尿嘧啶的标准曲线范围0.02~1.0μg·mL-1(r=0.9996),方法的重现性、回收率、精密度和稳定性均符合生物样品分析要求.卡培他滨代谢物

  6. The effect of annealing temperature on the structure and optical properties of sol-gel derived nanocrystalline cobalt aluminate spinel

    Kurajica, S., E-mail: [University of Zagreb, Faculty of Chemical Engineering and Technology, 19 Marulic Square, HR-10000 Zagreb (Croatia); Popovic, J. [Division of Materials Physics, Ruder Boskovic Institute, Bijenicka 54, Zagreb (Croatia); Tkalcec, E. [University of Zagreb, Faculty of Chemical Engineering and Technology, 19 Marulic Square, HR-10000 Zagreb (Croatia); Grzeta, B. [Division of Materials Physics, Ruder Boskovic Institute, Bijenicka 54, Zagreb (Croatia); Mandic, V. [University of Zagreb, Faculty of Chemical Engineering and Technology, 19 Marulic Square, HR-10000 Zagreb (Croatia)


    The thermal evolution of a sol-gel derived cobalt aluminate (CoAl{sub 2}O{sub 4}) of a spinel-type structure was studied. The samples were prepared by the sol-gel technique using aluminum-sec-butoxide [Al(O{sup s}Bu){sub 3}] and cobalt nitrate hexahydrate Co(NO{sub 3}){sub 2} Multiplication-Sign 6H{sub 2}O as starting materials and ethyl-acetoacetate (C{sub 6}H{sub 10}O{sub 3}) as a chelating agent. The powder precursors were annealed at various temperatures ranging from 200 to 900 Degree-Sign C. The properties of products were characterized using differential thermal analysis and thermo-gravimetric analysis (DTA/TGA), UV-vis diffuse reflectance spectroscopy (DRS) and X-ray diffraction (XRD). The UV-vis DRS spectra revealed that in samples annealed at temperatures below 700 Degree-Sign C a part of Co existed in a 3 + oxidation state. The accommodation of Co{sup 3+} in octahedral cation sites yields with a green color. Thermal treatment at higher temperatures promote the reduction of Co{sup 3+} ions to Co{sup 2+} and a change of color from green to blue, later being characteristic of Co{sup 2+} ions in tetrahedral coordination. The XRD revealed that annealing of the precursor powder at a lower temperature (500 and 600 Degree-Sign C) resulted in a crystal spinel-type phase Co{sub 2}AlO{sub 4.} Annealing at higher temperatures (700-900 Degree-Sign C) induces a reaction of Co{sub 2}AlO{sub 4} with amorphous alumina producing a phase CoAl{sub 2}O{sub 4} having a partially inverse spinel structure, with {delta} in the range of 0.198-0.260. The spinel powders obtained at all annealing temperatures were in the nanosize range. -- Graphical abstract: UV-vis DRS spectra of cobalt aluminate powders heat treated at various temperatures. Highlights: Black-Right-Pointing-Pointer Sol-gel derived spinel-type CoAl{sub 2}O{sub 4} thermal evolution was monitored. Black-Right-Pointing-Pointer Below 700 Degree-Sign C part of Co exists in 3{sup +} state (octahedral sites), yielding green

  7. 反向高效液相色谱测定法同时测定人血浆中5种镇静催眠药浓度%Simultaneous determination of 5 sedative hypnotics in human plasma by reversed phase high-performance liquid chromatography

    梁湘辉; 戴智勇; 刘文恩


    目的:建立人血浆中地西泮、硝西泮、奥沙西泮、艾司唑仑和阿普唑仑的反向高效液相色谱测定法(RP-HPLC).方法:以血浆为基质,配制含有5种镇静催眠药的混合标准品,取1 mL样品加入50 mg/L卡马西平10μL为内标,乙酸乙酯萃取后用100 μL流动相溶解,进样20 μL,通过高效液相色谱仪检测人血浆基质中5种镇静催眠药浓度.色谱条件:色谱柱为C18柱(4.6 mm×250 mm),柱温30℃,流动相为甲醇:水(65∶35),流速1.0 mL/min,紫外检测波长为230 nm.结果:地西泮、硝西泮、奥沙西沣、艾司唑仑和阿普唑仑在5~1 200μg/L范围内线性关系良好(r≥0.9966,P<0.05),加样回收率在95.5%~105.6%,萃取回收率均在75%以上,日内及日间变异相对标准偏差(RSD)均小于10%(n=5).结论:RP-HPLC方法简便、准确、灵敏,适用于人血浆中地西泮、硝西泮、奥沙西泮、艾司唑仑和阿普唑仑的血药浓度的同时检测.%Objective To determine diazepam, nitrazepam, oxazepam, estazolam, and al-prazolam simultaneously in human plasma by reversed phase high-performance liquid chromatography (RP-HPLC). Methods Ten microliter carbamazepine (50 mg/L)as the internal standard was added into 1 mL sample, which contained the 5 mixed sedative hypnotics as standard substance and human plasma as ground substance. They were extracted with acetoacetate from plasma samples, and then were dissolved by 100 μL mobile phase. The blood drug levels were analyzed by high perform-ance liquid chromatograph with 20 μL sample injection on a chromatographic column C 18 (4.6 mm×250 mm)at 30℃. The mobile phase consisted of methanol and water (65:35) , and the flow rate was 1.0 mL/min. The ultraviolet detection wavelength was 230 nm. Results The linearity range of the 5 drugs was 5~1 200 μg/L(r≥0.9966, P<0.05). The recovery rate was 95.5%~105.6%. The extraction recovery rate was more than 75%. The relative standard deviation (RSD) of intra-day and

  8. 1-酰基苯并咪唑酮酰胺衍生物的合成及其抗菌活性%Synthesis and antimicrobial activity of 1-acyl benzimidazolone amide derivatives

    李方方; 魏少鹏; 宗兆锋; 姬志勤


    以邻苯二胺和乙酰乙酸乙酯为起始原料制得异丙烯基苯并咪唑酮(Ⅲ),再经N-酰化反应得到13个苯并咪唑酮酰胺衍生物(Ⅳ-01~Ⅳ-13)以及由Ⅳ-02脱异丙烯基的产物Ⅳ-02a,其中9个为未见文献报道的新化合物.通过核磁共振氢谱和碳谱、质谱以及元素分析对其结构进行了表征.抑菌活性测定结果表明,化合物Ⅳ-01~Ⅳ-03、Ⅳ-11及Ⅳ-02a对供试病原细菌和真菌均表现出明显的抑菌活性,其中化合物Ⅳ-02和Ⅳ-02a尤为突出,且二者活性相近,其中Ⅳ-02对蜡状芽孢杆菌Bacillus cereus(1.184 6)、枯草芽孢杆菌Bacillus subtilis(1.88)、金黄色葡萄球菌Staphylococcus aureus (1.89)和大肠杆菌Escherichia coil(1.157 4)的MIC(抑制生长的最低浓度)值分别为0.78、12.5、1.56和1.56 μg/mL,对番茄灰霉病菌Botrytis cinerea的有效抑制中浓度(EC5o)为7.02 μg/mL.%Thirteen benzimidazolone derivatives were synthesized from isopropenyl-benzimidazolone via N-acylation reaction, and o-phenylenediamine and acetoacetic ester were used as starting material. The structures of the new derivatives were confirmed by 1H NMR,13C NMR and ESI-MS spectral analysis. Antimicrobial activity test indicated that compounds Ⅳ-01~Ⅳ-03 and Ⅳ-11,as well as the de-isopropenyl product of Ⅳ-02,Ⅳ-02a,have good inhibitory activity against the tested microbial.The MIC values of compound Ⅳ-02 were 0.78,12. 5,1. 56 and 1. 56 μg/mL against Bacillus cereus, Bacillus subtilis, Staphylococcus aureus and Escherichia coil,and the EC50 value was 7. 02 μg/mL against Botrytis cinerea, respectively. The de-isopropenyl product, Ⅳ-02a, showed the similarly antimicrobial activity as that of Ⅳ-02.

  9. Experimental Design for Green Chemistry Directed Hantzsch Reaction%绿色化学导向的Hantzsch反应实验设计

    查正根; 郑媛; 郑小琦; 汪志勇


    在水相中,70℃时,六亚甲基四胺、乙酰乙酸乙酯与碳酸铵发生Hantzsch反应,生成1,4-二氢-2,6-二甲基吡啶-3,5-二羧酸二乙酯,产率达90%;在55~60℃时,醛、乙酰乙酸乙酯和碳酸铵Hantzsch反应得到对称的Hantzsch酯,反应时间1~4 h,产率为88%~99%.以醋酸铵代替碳酸铵,在绿色有机溶剂(水、乙醇)中,催化剂(脯氨酸、胍盐酸盐)催化该反应得到中等至定量的产率;以5,5-二甲基-1,3-环己二酮代替另一分子乙酰乙酸乙酯,反应产生不对称的Hantzsch酯.该反应甚至在无溶剂、无催化剂、室温条件下,主要得到空气氧化的芳构化产物.以绿色化学为导向,将Hantzsch反应基础研究的成果设计成设计型、综合型实验,合成在化学、生物和医学上有广泛应用的Hantzsch酯,既开拓了学生的创新能力,培养了他们绿色环保意识,又理论联系实际、学以致用.%In aqueous media, Hantzsch reaction of hexamethylenetetramine and ethyl acetoacetate with ammonium carbonate occured to diethyl 1,4-dihydro-2,6-dimethylpyridine-3,5-dicarboxylate,in yield of 90% at 70 ℃.In 55 - 60℃ aldehyde and ethyl allylacetoacetate reacted with ammonium carbonate to give symmetrical Hantzsch ester in yield of 88% - 99% in 1 - 4 h.Instead of ammonium carbonate with ammonium acetate, in green organic solvent (water or ethanol),catalyst (proline or guanidine hydrochloride) catalyzed Hantzsch reaction in medium to quantitative yield.Instead of another ethyl allylacetoacetate with 5,5-dimethyl-cyclohexane-1, 3-dione, asymmetric Hantzsch reactions occured.Even without solvent and catalyst at ambient temperature, Hantzsch esters of aromatization were produced.With the green chemistry oriented, the basic research results of Hantzsch reaction were designed into were designed and comprehensive synthesis experiments to achieve Hantzsch esters, and they are widely used in chemistry, biology and medicine.Not only do they develop the

  10. Inhibition of Fatty Acid Synthase in vitro by Xuezhiling Tablets%血脂灵片对脂肪酸合酶的体外抑制作用

    王鹏; 高敏艳; 高岚; 肖学凤


    Objective: To study the inhibition of fatty acid synthase ( FAS) by Xuezhiling tablets in vitro, and explore its mechanism of action. Method: The ultraviolet spectroscopy was used to evaluate the activity of FAS through monitoring the alteration of absorbance ( A) value of reduced nicotinamide-adenine dinucleotide phosphate ( NADPH ) and the substrates were respectively acetyl coenzyme A ( AcCoA ) , malonyl-CoA (MalCoA) , acetoacetyl-coenzymeA ( AcAcCoA) , ethyl acetoacetate, ethyl crotonate, NADPH for different active sites. The inhibition of FAS by overall reduction and different active sites was separately detected after the treatment with different doses of Xuezhiling tablets. Result: The activity of FAS could be inhibited by Xuezhiling tablets. After the treatment with Xuezhiling tablets (150 mg·L-1 ) , the inhibition ratio was 51% with manner if time and dose relationship. For the different active sites of FAS, Xuezhiling tablets displayed different abilities. It showed more potential in inhibiting the enoyl reduction and AcAcCoA reduction than keto-acyl reduction, and it was also dose dependent for the different active sites. The residual activity of different active sites was less than 70% , and more than 50% , when FAS was treated with Xuezhiling tablets at the concentration of 150 mg ·L-1 . Conclusion: FAS can be inhibited by Xuezhiling tablets, and this ability was attributed to inhibit the different active sites of FAS. This article prove that the ratiocination about the lowering blood lipids of Xuezhiling tablets is related to FAS and it can be a reference for Xuezhiling tablets used for obesity and other disease that related to FAS.%目的:研究血脂灵片对脂肪酸合酶(fatty acid synthase,FAS)的体外抑制作用,并初步探索其作用机制.方法:以乙酰辅酶A(AcCoA),丙二酰辅酶A(MalCoA),乙酰乙酰辅酶A(AcAcCoA),乙酰乙酸乙酯,丁烯酸乙酯,原型辅酶Ⅱ(NADPH)等为底物,采用紫外分光光度法,通过测定340

  11. Ketone bodies metabolism during ischemic and reperfusion brain injuries following bilateral occlusion of common carotid arteries in rats Metabolismo dos corpos cetônicos durante as lesões de isquemia e reperfusão cerebrais após oclusão bilateral das artérias carótidas comuns em ratos

    Mário Henrique Girão Faria


    Full Text Available PURPOSE: To evaluate the in vivo alterations on ketone bodies metabolism after cerebral ischemia/reperfusion through an experimental model of brain ischemia induced by simple occlusion of common carotid arteries (CCAs in Wistar rats. METHODS: Forty-eight male Wistar rats were randomly distributed on two groups (S - Sham; T - Test and further redistributed into four times sets of study. After bilateral occlusion of CCAs for 30min, the animals of group T were allowed reperfusion for 0, 5, 10 and 15min. Samples of cerebral tissue and systemic arterial blood were collected and the metabolites acetoacetate (ACT and beta-hydroxybutyrate (BHB were determined. RESULTS: Cerebral ACT and BHB levels increased significantly in Group T after 30min of carotid occlusion (time 0. The highest brain ketone bodies (ACT+BHB concentration was verified at 5min of reperfusion, decreasing after 10min of recirculation. Systemic ketone bodies levels increased similarly between test and sham groups. Group S demonstrated a significant increase in cerebral and systemic ACT and BHB concentrations mainly after 40-45min of study. CONCLUSIONS: The partial transient acute global brain ischemia induced by the bilateral carotid occlusion in Wistar rats triggered ketogenesis probably due to a central stimulation of catecholamine secretion. There was an increased cerebral uptake of ketone bodies following brain ischemia, reaffirming these metabolites as alternative energy substrates under conditions of cerebral metabolic stress as well as its potential role on neuroprotection. The greatest changes in ketone bodies metabolism were verified at initial minutes of recirculation as a result of the reperfusion injury phenomenon.OBJETIVO: Avaliar as alterações in vivo no metabolismo dos corpos cetônicos após isquemia/reperfusão cerebral através de um modelo experimental de isquemia cerebral induzido pela simples oclusão das artérias carótidas comuns (CCAs em ratos Wistar. M

  12. Observation of curative effect of modified atkins diet in treatment of adults with refractory epilepsy%改良的阿特金斯饮食治疗成人难治性癫痫的疗效观察

    尚丽; 王曼; 龚德山; 张翠荣; 王开颜


    目的:评估改良的阿特金斯饮食( MAD)治疗成人难治性癫痫的疗效。方法19例成人难治性癫痫患者接受MAD治疗,随访1~15个月。记录发作频率及不良反应。结果1个月有效率为63%,保留率100%;3个月有效率57%,保留率89%;6个月有效率为47%,保留率为47%。患者治疗前发作频率为4~152次/月,平均25次/月;治疗后1个月发作频率为0~100次/月,平均8次/月。住院期间患者β-羟丁酸(1.5±0.6)mmol/L,血糖(4.6±0.4)mmol/L;随访期间乙酰乙酸波动于触~揣。与治疗前比较,治疗后第1个月和第3个月三酰甘油水平显著升高(均P<0.05),但均值仍在正常范围;总胆固醇及低密度脂蛋白水平差异无统计学意义。不良反应表现为体质量减轻7例,高脂血症6例,乏力4例,便秘3例,腹泻2例,呕吐及肾结石1例。结论 MAD治疗成人难治性癫痫安全有效,没有手术指征的成人难治性癫痫患者排除禁忌后可选择MAD。%Objective To evaluate the efficacy of modified atkins diet ( MAD ) in treatment of adults with refractory epilepsy.Methods Nineteen adult patients with refractory epilepsy were initiated on the MAD and followed 1-15 months.Seizure frequencies and adverse effects were recorded.Results The effective rate was 63%, and retention rate was 100%at 1 month after MAD treatment.The effective rate was 57%, and retention rate was 89%at 3 month after MAD treatment.The effective rate was 47%, and retention rate was 47% at 6 month after MAD treatment.The seizure frequencies was 4-152 times per month before treatment, and the average was 25 times per month;the seizure frequencies was 0-100 times per month at 1 month after treatment, and average was 8 per month. The serumβ-hydroxybutyric acid level was (1.5 ±0.6)mmol/L, blood glucose level was (4.6 ±0.4)mmol/L in the hospitalization period;the level of acetoacetic acid

  13. Effects of ketogenic amino acid replacement diet on insulin resistance in mice fed with high fat diet%富生酮氨基酸饮食对高脂诱导的小鼠胰岛素抵抗的影响

    徐玲; 马红艳; 李佳; 高陈林; 徐勇


    目的:探讨富生酮氨基酸饮食对高脂诱导小鼠胰岛素抵抗的影响及机制。方法 C57BL小鼠随机分4组,给予常规饮食( Con)、高脂饮食( HFD)、富生酮氨基酸高脂饮食( HFDKAAR )以及高脂喂养8周后改为富生酮氨基酸高脂饮食( HFD→HFDKAAR )喂养。测食量、体重及内脏脂肪,16周后行腹腔内葡萄糖耐量试验,测血糖、胰岛素、血酮,根据血糖和胰岛素计算曲线下面积( AUC)及胰岛素抵抗指数( IRI),检测肝脏LKB1、AMPK、mTOR蛋白和mcp-1 mRNA表达。结果各组小鼠摄入量及血酮无统计学差异。与Con组比较,HFD组小鼠表现体重及内脏脂肪增加、糖耐量受损和高胰岛素水平,肝LKB1、p-AMPK表达下降,p-mTOR蛋白和mcp-1 mRNA表达增加。与HFD比较,HFDKAAR及HFD→HFDKAAR组小鼠体重及内脏脂肪均减轻,血糖、AUC、胰岛素浓度和IRI均降低(P<0.05),肝LKB1、p-AMPK、p-mTOR蛋白和mcp-1 mRNA表达异常均被明显逆转(P<0.05)。结论富生酮氨基酸饮食可能通过调节肝脏LKB1-AMPK-mTOR通路,降低mcp-1 mRNA水平,改善或逆转高脂饮食诱导的肥胖、胰岛素抵抗及糖耐量受损。%Objective To investigate the effects of ketogenic amino acid ( KAA) replacement diet on insulin resistance in mice fed with high fat diet(HFD) and to analyze the possible mechanism. Methods C57BL mice were fed with a control diet, HFD, and KAA-fortified HFD(HFDKAAR)from the age of 8 weeks, and 8 weeks after HFD initiation, the HFD-fed mice were divided into two groups:one group of mice were fed the same HFD, the other group were fed HFDKAAR ( HFD→HFDKAAR ) . The metabolic evaluations were performed at the end of 16 weeks. Blood glucose levels were measured at 0, 15, 30, 60, 90, and 120 min after the injection of glucose ( 1 g/kg BW intraperitoneal glucose tolerance test, ipGTT) . The insulin,β-hydroxybutyrate, and acetoacetate levels in the plasma were measured via ELISA. The insulin resistance index ( IRI) and

  14. No-mediated mitochondria damage induce liver function failure in rats sepsis%一氧化氮介导的线粒体损害诱发败血症肝功衰竭的研究

    涂巍; 曲文志; 赵嫚; 于作夫; 金光华; 王勤勇; 胡松


    nucleotide in the hepatocytes with the high-performance liquid chromatography ( HPLC),and determined the content of the ketone body in the hepatocytes of the two groups with the enzymatic method,then calculated the ketone body ratio ( acetoacetate/β-hydroxy butyrate,KBR).Results The production of NO in the CLP group is twice to third as many as that in the Sham group ,IL-1β can decrease the content of ATP and the KBR in the hepatocytes of the two groups,and the decrease degree in the CLP group is greater than that in the Sham group.When the L-arginine was injected,the production of NO in the hepatocytes in the CLP group is increased,and the level of ATP and KBR is decreased.NG-methyl-L-arginine (L-NMMA),the inhibitor of NO syntbetase,can inhibit the production of NO,and recuperate the decrease of ATP and KBR.Conclusion Enhanced production of NO primed by the septic response may be associated with hepatic dysfunction in the liver of septic rats.The regulation of the production of NO may be an effective method in the prevention of the hepatic dysfunction in sepsis.

  15. Effect of monomers extracted from Spatholobus suberectus Dunn on proliferation of hematopoietic progenitor cells in marrow-depressed mice%鸡血藤单体化合物对骨髓抑制小鼠造血祖细胞增殖的影响

    王东晓; 刘屏; 陈若芸; 陈孟莉; 陈桂芸


    BACKGROUND: Hematogenesis of a body mainly depends on the proliferation of hematopoietic progenitor cell (HPC). Hematopoietic functional impairment will occur when hematopoietic cells are injured by radioactive ray or chemical drug. The proliferation of HPC is the key link of promoting hematogenesis.OBJECTIVE: To study the effects of nine monomers extracted from Spatholobus suberectus Dunn (SSD) on proliferation of HPC in marrow-depressed mice. DESIGN: Randomized controlled trial.SETTING: Department of Pharmacy, General Hospital of Chinese PLA.MATERIALS: This experiment was conducted in the Department of Clinical Pharmacology, General Hospital of Chinese PLA from November 2002 to February 2003. Totally 348 healthy Kunming mice, weighing 22-25g, clean grade, of irrespective gender, were selected in this study (certification: SCXK-2001-001). The animal experiment was approved by the local ethics committee. SSD was provided by Dispensary of Traditional Chinese Medicine, General Hospital of Chinese PLA; monomers (gallocatechin, formononetin, catechin, pyromucic acid, syringic acid, Demethylvestitol, 1,3,5-benzenetriol, ononin, and epicatechin) were extracted from SSD acetoacetate; TGL-16 centrifuger was made in Shanghai 6th Medical Equipment Factory; CO2 incubator was made in SANYO Company, Japan; MK inverted microscope was provided by OLYMPUS Company, Japan.METHODS: Experimental grouping: Mice were randomly divided into 29 groups, including normal group; control group; gallocatechin high-, medium-, low-dose groups; formononetin high-, medium-, low-dose groups; catechin high-, medium-, low-dose groups; pyromucic acid high-, medium-, low-dose groups; syringic acid high-, medium-, low-dose groups; Demethylvestitol high-, medium-, low-dose groups; 1,3,5-benzenetriol high-, medium-, low-dose groups; ononin high-, medium-, low-dose groups; epicatechin high-, medium-, low-dose groups with 12 mice in each group. Experimental intervention: All the mice except the mice in