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Sample records for acetoacetates

  1. 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.

    2005-01-01

    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…

  2. [11C]-Acetoacetate PET imaging: a potential early marker for cardiac heart failure

    The ketone body acetoacetate could be used as an alternate nutrient for the heart, and it also has the potential to improve cardiac function in an ischemic–reperfusion model or reduce the mitochondrial production of oxidative stress involved in cardiotoxicity. In this study, [11C]-acetoacetate was investigated as an early marker of intracellular damage in heart failure. Methods: A rat cardiotoxicity heart failure model was induced by doxorubicin, Dox(+). [14C]-Acetoacetate, a non-positron (β −) emitting radiotracer, was used to characterize the arterial blood input function and myocardial mitochondrial uptake. Afterward, [11C]-acetoacetate (β +) myocardial PET images were obtained for kinetic analysis and heart function assessment in control Dox(−) (n = 15) and treated Dox(+) (n = 6) rats. The uptake rate (K1) and myocardial clearance rate (k2or kmono) were extracted. Results: [14C]-Acetoacetate in the blood was increased in Dox(+), from 2 min post-injection until the last withdrawal point when the heart was harvested, as well as the uptake in the heart and myocardial mitochondria (unpaired t-test, p < 0.05). PET kinetic analysis of [11C]-acetoacetate showed that rate constants K1, k2 and kmono were decreased in Dox(+) (p < 0.05) combined with a reduction of 24% of the left ventricular ejection fraction (p < 0.001). Conclusion: Radioactive acetoacetate ex vivo analysis [14C], and in vivo kinetic [11C] studies provided evidence that [11C]-acetoacetate can assess heart failure Dox(+). Contrary to myocardial flow reserve (rest–stress protocol), [11C]-acetoacetate can be used to assess reduced kinetic rate constants without requirement of hyperemic stress response. The proposed [11C]-acetoacetate cardiac radiotracer in the investigation of heart disease is novel and paves the way to a potential role for [11C]-acetoacetate in cardiac pathophysiology

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

    Adi Wolfson; Nisim Haddad; Chrstina Dlugy; Dorith Tavor; Yoram Shotland

    2008-01-01

    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

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

    Adi Wolfson

    2008-09-01

    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

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

    Sols obtained by chelation of titanium n-butoxide with ethyl acetoacetate, Eaa, in various ratios have been subjected to FTIR, 1H and 13C 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(OnBu)2(Eaa)2, bis-chelated dimer, (Ti(OnBu)3Eaa)2 and monochelated dimer, Ti2(OnBu)7Eaa have been established. As the molar ratio Eaa/Ti(OnBu)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 Ti2(OnBu)7Eaa 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

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

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

    2012-01-01

    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...

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

    Kurajica, S., E-mail: stankok@fkit.hr; Škorić, I.; Lozić, I.; Mandić, V.

    2014-10-15

    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.

  8. Sbi00515, a Protein of Unknown Function from Streptomyces bingchenggensis, Highlights the Functional Versatility of the Acetoacetate Decarboxylase Scaffold.

    Mueller, Lisa S; Hoppe, Robert W; Ochsenwald, Jenna M; Berndt, Robert T; Severin, Geoffrey B; Schwabacher, Alan W; Silvaggi, Nicholas R

    2015-06-30

    The acetoacetate decarboxylase-like superfamily (ADCSF) is a group of ~4000 enzymes that, until recently, was thought to be homogeneous in terms of the reaction catalyzed. Bioinformatic analysis shows that the ADCSF consists of up to seven families that differ primarily in their active site architectures. The soil-dwelling bacterium Streptomyces bingchenggensis BCW-1 produces an ADCSF enzyme of unknown function that shares a low level of sequence identity (~20%) with known acetoacetate decarboxylases (ADCs). This enzyme, Sbi00515, belongs to the MppR-like family of the ADCSF because of its similarity to the mannopeptimycin biosynthetic protein MppR from Streptomyces hygroscopicus. Herein, we present steady state kinetic data that show Sbi00515 does not catalyze the decarboxylation of any α- or β-keto acid tested. Rather, we show that Sbi00515 catalyzes the condensation of pyruvate with a number of aldehydes, followed by dehydration of the presumed aldol intermediate. Thus, Sbi00515 is a pyruvate aldolase-dehydratase and not an acetoacetate decarboxylase. We have also determined the X-ray crystal structures of Sbi00515 in complexes with formate and pyruvate. The structures show that the overall fold of Sbi00515 is nearly identical to those of both ADC and MppR. The pyruvate complex is trapped as the Schiff base, providing evidence that the Schiff base chemistry that drives the acetoacetate decarboxylases has been co-opted to perform a new function, and that this core chemistry may be conserved across the superfamily. The structures also suggest possible catalytic roles for several active site residues. PMID:26039798

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

    Quadros Edward V

    2009-05-01

    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.

  10. New liner and reticular polyelectrolytes based an acetoacetic ehter and acrylic acid: synthesis, structure and stimulating sensitivity characteristics

    Novel linear and crosslinked poly-carbolytes based on acetoacetic ester and acrylic acid were synthesized for the first time via Michael addition reaction followed by the radical poly-mineralization. The structure and properties of polyelectrolytes were characterized by FTIR, Raman and NMR spectroscopy, TGA, DSC, GPC, viscosimetry, and potentiometric titration. Considerable influence of water content on the swelling rate of hydrogel was established. The swelling-deswelling properties of hydrogels were studied in water-organic solvent mixtures, electric and magnetic fields. (author)

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

    Laishun Shi

    2012-01-01

    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.

  12. 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

    1997-01-01

    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,

  13. 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.

    1997-01-01

    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

  14. Importance of Time Scale and Local Environment in Electron-Driven Proton Transfer. The Anion of Acetoacetic Acid.

    Keolopile, Zibo G; Gutowski, Maciej; Buonaugurio, Angela; Collins, Evan; Zhang, Xinxing; Erb, Jeremy; Lectka, Thomas; Bowen, Kit H; Allan, Michael

    2015-11-18

    Anion photoelectron spectroscopy (PES) and electron energy-loss spectroscopy (EELS) probe different regions of the anionic potential energy surface. These complementary techniques provided information about anionic states of acetoacetic acid (AA). Electronic structure calculations facilitated the identification of the most stable tautomers and conformers for both neutral and anionic AA and determined their relative stabilities and excess electron binding energies. The most stable conformers of the neutral keto and enol tautomers differ by less than 1 kcal/mol in terms of electronic energies corrected for zero-point vibrations. Thermal effects favor these conformers of the keto tautomer, which do not support an intramolecular hydrogen bond between the keto and the carboxylic groups. The valence anion displays a distinct minimum which results from proton transfer from the carboxylic to the keto group; thus, we name it an ol structure. The minimum is characterized by a short intramolecular hydrogen bond, a significant electron vertical detachment energy of 2.38 eV, but a modest adiabatic electron affinity of 0.33 eV. The valence anion was identified in the anion PES experiments, and the measured electron vertical detachment energy of 2.30 eV is in good agreement with our computational prediction. We conclude that binding an excess electron in a π* valence orbital changes the localization of a proton in the fully relaxed structure of the AA(-) anion. The results of EELS experiments do not provide evidence for an ultrarapid proton transfer in the lowest π* resonance of AA(-), which would be capable of competing with electron autodetachment. This observation is consistent with our computational results, indicating that major gas-phase conformers and tautomers of neutral AA do not support the intramolecular hydrogen bond that would facilitate ultrarapid proton transfer and formation of the ol valence anion. This is confirmed by our vibrational EELS spectrum. Anions

  15. 附子乙酸乙酯提取物对虚寒模型大鼠肝LA、LDH、PA、Gn含量及ATP酶活力的影响%Effects of Acetoacetate Extract of Radix Aconite on Hepatic Contents of LA, LDH, PA, Gn, and ATPase Activities in Deficient Cold Model Rats

    刘一洋; 王世军; 韩冰冰; 刘珊; 杨富梅

    2011-01-01

    目的 观察中药附子乙酸乙酯提取物与附子水煎液相比对虚寒模型大鼠能量代谢的影响.方法 将Wistar大鼠随机分为空白对照组(空白组,10只)及虚寒模型组(30只).采用生石膏、龙胆草、黄柏和知母水煎液灌胃大鼠15天复制虚寒模型,之后将虚寒模型组大鼠再随机分为附子乙酸乙酯提取物组(治疗组,10只)、附子水煎液组(对照组,10只)及模型组(10只),两个用药组分别用附子乙酸乙酯提取物和附子水煎液分别灌胃治疗5天,模型组继续予模型药灌胃,空白对照组继续予生理盐水灌胃.检测大鼠肝组织乳酸(LA)、乳酸脱氢酶(LDH)、丙酮酸(PA)、糖原(Gn)含量以及Na+ -K+ -ATP酶、Ca2+ -Mg2+ -ATP酶的活力.结果 与空白组比较,模型组PA含量、Na+ -K+ -ATP酶、Ca2+ -Mg2+ -ATP酶活力降低;与模型组比较,两个给药组PA含量升高;与对照组比较,治疗组LDH含量、PA含量、Na+ -K+ -ATP酶活力升高,差异均有统计学意义(P<0.05).结论 附子乙酸乙酯提取物比附子水煎液热性略大,似乎能产生更多的能量,但是最终结论及具体作用机制有待进一步研究.%Objective To observe different effects of acetoacetate extract of Radix Aconite and Radix Aconite Decoction on the energy metabolism in deficient cold model rats. Methods Wistar rats were randomly divided into the blank control group (n = 10) and the deficient cold model group (n = 30). The deficient cold rat model was established using decoction consisting of gypsum, Radix Gentianae, Cortex Phellodendri, and Rhizoma Anemarrhenae. The decoction was given to rats of the deficient cold model group by gastrogavage for 5 days. Then these rats were randomly divided into the acetoacetate extract of Radix Aconite group (n = 10), the Radix Aconite Decoction group (n = 10), and the model group (n = 10). Rats in the model model group were administered with the decoction by gastrogavage. Rats in the other two groups were

  16. A convenient synthesis of 3-aryl-1,2,4-oxadiazoles from ethyl acetoacetate and amidoximes under solvent-free conditions

    Juliana L. L. F. Regueira

    2012-06-01

    Full Text Available 1,2,4-Oxadiazole containing compounds have attracted great attention due to their applications in material chemistry and therapeutics. Herein we report a convenient synthesis of 3-aril-[1,2,4-oxadiazol-5yl] propan-2-one (3a-d by heating of b-ceto esters and an amidoximes without any solvent and in the absence of base. The arylamidoximes (1a-f was synthesized in moderate and excellent yields (31- 89% by reaction of nitrile with hydroxylamine hydrochloride in water at 25ºC. In the general synthetic strategy employed in our study is illustrated in scheme 1. The 3-aril-[1,2,4-oxadiazol-5yl] propan-2-one (3a-f were synthesized by treatment of arylamidoxime (1a-f with b-ceto esters for 4 hour without any solvent and in the absence of base. The reaction was monitored  by TLC   (thin layer chromatography. The heterocycles (3a-f were obtained in moderate and good yields (60-88%. The products were identified using both analytical and spectral data (IR, 1H and 13C NMR and all compounds are in full agreement with the proposed structure.

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

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

    2010-01-01

    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...

  18. NCBI nr-aa BLAST: CBRC-CINT-01-0205 [SEVENS

    Full Text Available CBRC-CINT-01-0205 ref|ZP_01855183.1| acetoacetate metabolism regulatory protein ATOC [Plancto...myces maris DSM 8797] gb|EDL59033.1| acetoacetate metabolism regulatory protein ATOC [Planctomyces maris DSM 8797] ZP_01855183.1 6.6 35% ...

  19. Synthesis and investigation of solvent effects on the ultraviolet absorption spectra of 5-substituted-4-methyl-3-cyano-6-hydroxy-2-pyridones

    NATASA V. VALENTIC

    2001-08-01

    Full Text Available A number of 5-substituted-4-methyl-3-cyano-6-hydroxy-2-pyridones from cyanoacetamide and the corresponding alkyl ethyl acetoacetates were synthesized according to modified literature procedures. The alkyl ethyl acetoacetates were obtained by the reaction of C-alkylation of ethyl acetoacetate. An investigation of the reaction conditions for the synthesis of 4-methyl-3-cyano-6-hydroxy-2-pyridone from cyanoacetamide and ethyl acetoacetate in eight different solvents was also performed. The ultraviolet absorption spectra of synthesized pyridones were measured in nine different solvents in the range 200–400 nm. The effects of solvent polarity and hydrogen bonding on the absorption spectra are interpreted by means of linear solvation energy relationships using a general equation of the form n = n0 + sp* + aa + bb, where p* is a measure of the solvent polarity, a is the scale of the solvent hydrogen bond donor acidities and b is the scale of the solvent hydrogen bond acceptor basicities.

  20. SYNTHESIS AND MICROBIAL EVALUATION OF NOVEL TETRAZOLO-TRIAZOLE DERIVATIVES SYNTHESIS und mikrobielle Evaluierung neuartiger Tetrazolo-triazol DERIVATE

    Vijay V Dabholkar, Navnath Shinde, Abhishekh Karekar and Prem Naik

    2013-01-01

    Full Text Available 5-(4'-Bromomethyl-1, 1'-biphenyl-2-yl -1H-tetrazole (1 was converted into its azide derivative using sodium azide (2, which on further treatment with malonitrile, diethyl malonate, ethyl acetoacetate and isopropyl acetoacetate to yielded the respective tetrazole-triazole derivatives. The structures of the synthesized compounds were confirmed by Physico-chemical test and spectral techniques, representative samples were screened for their antimicrobial activity against gram positive and gram negative bacteria. Key words: Tetrazoles, Triazoles, Triphenyl phosphine, azides

  1. Nano aluminium nitride as a solid source of ammonia for the preparation of Hantzsch 1,4-dihydropyridines and bis -(1,4-dihydropyridines) in water via one pot multicomponent reaction

    Ghorbani-Choghamarani, Arash; Goudarziafshar, Hamid; Nikoorazm, Mohsen; Yousefi, Somaieh; Tahmasbi, Bahman, E-mail: arashghch58@yahoo.co [Ilam Univ., Ilam (Iran, Islamic Republic of). Faculty of Science. Dept. of Chemistry; Zolfigol, Mohammad Ali; Hajjami, Maryam [Bu-Ali Sina University, Hamadan (Iran, Islamic Republic of). Faculty of Chemistry

    2011-07-01

    Nano aluminium nitride in the presence of water acts as solid source of ammonia, which is used for the preparation of 1,4-dihydropyridines and bis-(1,4-dihydropyridines). An efficient and simple procedure for the one-pot synthesis of 1,4-dihydropyridine and bis-(1,4-dihydropyridine) derivatives was achieved by combination of methyl acetoacetate or ethyl acetoacetate with aldehydes or dialdehydes and aluminium nitride at 80 deg C in water in high purity and good yields. (author)

  2. Pechmann Reaction Promoted by Boron Trifluoride Dihydrate

    J. Mezger

    2005-08-01

    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 %.

  3. 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

    2016-04-01

    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.

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

    Khazaei, Marziyeh; Anary-Abbasinejad, Mohammad; Hassanabadi, Alireza; Sadeghi, Bahareh

    2012-01-01

    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.

  5. New Condensation Reaction of β-keto-δ-valerolactones, Carbon Disulfide and Alkyl Halides

    You Ming WANG; Yu Xin LI; Su Hua WANG; Zheng Ming LI

    2004-01-01

    β-Keto-δ-valerolactones, which were obtained by reaction of acetoacetate with aldehydes or ketones, reacted with carbon disulfide, alkyl halides and a new condensation reaction was developed. The structures of the products 3 were confirmed by 1HNMR spectra and elemental analysis.

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

    Jalal Albadi

    2014-10-01

    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.

  7. The heterocyclic compounds on the base of glycerin derivatives

    On the base of functional glycerin derivatives as a result of condensation them with different compounds (aldehydes, ketones, acetoacetic ether and etc.) receive vast row of heterocyclic compounds, which found wide use as a drugs in medicine, as a insects-fungicides in agriculture, as a monomers for polymerization, as a reagents for organic synthesis and etc

  8. 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

    2012-07-01

    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.

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

    Heravi, Majid M; Daraie, Mansoureh

    2016-01-01

    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. PMID:27043522

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

    Farahnaz Kargar Behbahani; Hourieh Sadat Alaei

    2013-05-01

    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.

  11. Direct C-H alkylation and indole formation of anilines with diazo compounds under rhodium catalysis.

    Mishra, Neeraj Kumar; Choi, Miji; Jo, Hyeim; Oh, Yongguk; Sharma, Satyasheel; Han, Sang Hoon; Jeong, Taejoo; Han, Sangil; Lee, Seok-Yong; Kim, In Su

    2015-12-18

    The rhodium(III)-catalyzed direct functionalization of aniline C-H bonds with α-diazo compounds is described. These transformations provide a facile construction of ortho-alkylated anilines with diazo malonates or highly substituted indoles with diazo acetoacetates. PMID:26458276

  12. Characterization of some Pr(III) complexes in terms of electronic spectral parameters

    Pr(III) complexes from the ligands derived from methyl acetoacetate, ethyl acetoacetate, veratraldehyde, ethyl vanillin and 2,5 dimethoxy benzaldehyde forming Schiff-bases with ortho, meta and para phenylene diamines have been synthesized. The complexes have been characterized in terms of various Slater-Condon Lande and Judd-Ofelt parameters. The various trends in the parametric values have also been described. The involvement of 4f-orbital in the Pr(III) complexes including deviation in the symmetry have been discussed on the basis of electronic spectral parameters. The validity of the theories used has been established while comparing observed and calculated energies and intensities of the various bands in the present complexes on the basis of r.m.s deviation. The trends of the curves observed in the solution spectra have also been discussed. (author). 21 refs., 5 tabs., 2 figs

  13. 西尼地平的合成工艺研究%Study on the Synthesis Technology of Cilnidipine

    施务务; 李立标

    2014-01-01

    With 2-methoxyethyl acetoacetate and cinnamyl acetoacetate as the starting materials, the cilnidipine can be obtained from 3-Aminocrotonic Cinnamate by Aldol condensation and Hantzsch cyclization. It is a suitable way to produce cilnidipin industrially,which provides the advantage of simplicity and safety.%以乙酰乙酸甲氧基乙基酯、乙酰乙酸肉桂醇酯为起始原料,经中间体3-氨基巴豆酸肉桂醇酯,通过Aldol缩合、Hantzsch环合反应,合成出了钙离子拮抗剂西尼地平。该合成工艺安全可行,操作简单,适合工业化生产。

  14. 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

    2012-03-01

    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.

  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

    2013-11-01

    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. Mechanism of Inhibition of Aliphatic Epoxide Carboxylation by the Coenzyme M Analog 2-Bromoethanesulfonate*

    Boyd, Jeffrey M.; Clark, Daniel D.; Kofoed, Melissa A.; Ensign, Scott A.

    2010-01-01

    The bacterial metabolism of epoxypropane formed from propylene oxidation uses the atypical cofactor coenzyme M (CoM, 2-mercaptoethanesulfonate) as the nucleophile for epoxide ring opening and as a carrier of intermediates that undergo dehydrogenation, reductive cleavage, and carboxylation to form acetoacetate in a three-step metabolic pathway. 2-Ketopropyl-CoM carboxylase/oxidoreductase (2-KPCC), the terminal enzyme of this pathway, is the only known member of the disulfide oxidoreductase fam...

  17. SODIUM SACCHARIN AS A CLEAN AND EFFICIENT CATALYST FOR THE SYNTHESIS OF 4-ARYLIDENE-3-METHYLISOXAZOL-5(4H-ONES VIA ONE-POT THREE-COMPONENT REACTION IN AQUEOUS MEDIUM. Saccharin-Natrium als saubere und effizienter Katalysator für die Synthese von 4-Aryliden-3-Methylisoxazol-5 (4H-onen über One-POT Dreikomponentenreaktion in wässrigem Medium.

    Hamzeh Kiyani*, Fatemeh Ghorbani

    2013-07-01

    Full Text Available As a result of one-pot three-component reaction of ethyl acetoacetate with hydroxylamine hydrochloride and various aromatic aldehydes using sodium saccharin as a catalyst in water, a green and environmentally benign solvent, 4-arylidene-3-methylisoxazol-5(4H-ones were obtained in high yields. The advantage of this method is efficient, clean, easy work-up, high yields, and shorter reaction time.

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

    Ghalem, Wassima; Boulcina, Raouf; Debache, Abdelmadjid

    2012-01-01

    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.

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

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

    2014-01-01

    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 a...

  20. Enhancement of L-3-hydroxybutyryl-CoA dehydrogenase activity and circulating ketone body levels by pantethine. Relevance to dopaminergic injury

    de Reggi Max; Nieoullon André; Khrestchatisky Michel; Abou-Hamdan Mhamad; Cornille Emilie; Gharib Bouchra

    2010-01-01

    Abstract Background The administration of the ketone bodies hydroxybutyrate and acetoacetate is known to exert a protective effect against metabolic disorders associated with cerebral pathologies. This suggests that the enhancement of their endogenous production might be a rational therapeutic approach. Ketone bodies are generated by fatty acid beta-oxidation, a process involving a mitochondrial oxido-reductase superfamily, with fatty acid-CoA thioesters as substrates. In this report, emphasi...

  1. 9-Ethoxy-1,5,13-trimethyl-8,10-dioxatetracyclo[7.7.1.02,7.011,16]heptadeca-2,4,6,11,13,15-hexaene

    Ewa Maria Nowakowska-Bogdan

    2009-09-01

    Full Text Available The reaction of ethyl acetoacetate with meta-cresol in an acidic ionic liquid yielded a complex mixture of condensation products. 4,7-Dimethylcoumarin and the title compound, C20H22O3, were isolated. The title compound shows chemical but not crystallographic mirror symmetry. The two aromatic rings are inclined at an angle of 73.55 (6°.

  2. Benzothiazolylazo derivatives of some β-dicarbonyl compounds and their Cu(II), Ni(II) and Zn(II) complexes

    KRISHNANNAIR KRISHNANKUTTY; MUHAMMED BASHEER UMMATHUR; DAMODARAN KAMALAKSHY BABU

    2010-01-01

    The coupling of diazotized 2-aminobenzothiazole with 1,3-dicar-bonyl compounds (benzoylacetone, methyl acetoacetate and acetoacetanilide) yielded a new series of tridentate ligand systems (HL). Analytical, IR, 1H--NMR, 13C-NMR and mass spectral data indicated that the compounds exist in the intramolecularly hydrogen bonded azo-enol tautomeric form in which one of the carbonyl groups of the dicarbonyl moiety had enolised and hydrogen bonded to one of the azo nitrogen atoms. The compounds forme...

  3. Synthesis of Polyfunctionalized 4H-Pyrans

    Manisha Bihani

    2013-01-01

    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.

  4. [Total synthesis of nordihydroguaiaretic acid].

    Wu, A X; Zhao, Y R; Chen, N; Pan, X F

    1997-04-01

    beta-Keto ester(5) was obtained from vanilin through etherification, oxidation and condensation with acetoacetic ester, (5) on oxidative coupling reaction by NaOEt/I2 produced dimer (6) in high yield. Acid catalyzed cyclodehydration of (6) gave the furan derivative(7), and by a series of selective hydrogenation nordihydroguaiaretic acid, furoguaiacin dimethyl ether and dihydroguaiaretic acid dimethyl ether were synthesized. PMID:11499030

  5. The Homogentisate Pathway: a Central Catabolic Pathway Involved in the Degradation of l-Phenylalanine, l-Tyrosine, and 3-Hydroxyphenylacetate in Pseudomonas putida

    Arias-Barrau, Elsa; Olivera, Elías R.; Luengo, José M.; Fernández, Cristina; Galán, Beatriz; García, José L.; Díaz, Eduardo; Miñambres, Baltasar

    2004-01-01

    Pseudomonas putida metabolizes Phe and Tyr through a peripheral pathway involving hydroxylation of Phe to Tyr (PhhAB), conversion of Tyr into 4-hydroxyphenylpyruvate (TyrB), and formation of homogentisate (Hpd) as the central intermediate. Homogentisate is then catabolized by a central catabolic pathway that involves three enzymes, homogentisate dioxygenase (HmgA), fumarylacetoacetate hydrolase (HmgB), and maleylacetoacetate isomerase (HmgC), finally yielding fumarate and acetoacetate. Wherea...

  6. NMR Studies on the Subacute Biochemical Effects of Aristolochic Acid on Rat Serum

    Xiao Yu ZHANG; Hui Feng WU; Xiao Jing LI; Feng Kui PEI; Jia Zuan NI

    2005-01-01

    The subacute effect of aristolochic acid (AA) on rat serum was studied by NMR method.The biochemical effects induced by AA were characterized by an increase in the amounts of creatinine, trimethylamine N-oxide, acetoacetate, acetate and 3-D-hydroxybutyrate and lactate in serum from 1H NMR spectra. Principal component analysis was used for further comparing the similarities of 1H NMR spectral profiles of serum from rats treated with AA and model toxins.

  7. Fatty acid oxidation and ketogenesis in astrocytes

    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 CO2 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 CO2 and labeled ketone bodies were produced from the oxidation of fatty acids labeled at carboxy- and ω-terminal carbons, indicating that fatty acids were oxidized by β-oxidation. The results from the radiolabeled tracer studies also indicated that a substantial proportion of the ω-terminal 4-carbon unit of the fatty acids bypassed the β-ketothiolase step of the β-oxidation pathway. The [14C]acetoacetate formed from the [1-14C]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 [14C]acetoacetate formed from the (ω-1)labeled fatty acids contained 90% of the label at carbon 3 and 10% at carbon 1

  8. 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.

    1987-11-01

    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 mumol.kg-1.min-1, 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.

  9. Validation of two-pool model for in vivo ketone body kinetics

    Previous studies have indicated that simultaneous infusions of two ketone body tracers ([13C]acetoacetate and [14C]beta-hydroxybutyrate) provide accurate estimates of exogenous ketone body inflow when an open two-pool model is employed. In the present studies, net hepatic ketone body production was determined from surgically placed arterial, portal venous, and hepatic venous catheters in conscious diabetic (n = 6) and 4-day fasted (n = 7) dogs. [13C]acetoacetate and [14C]beta-hydroxybutyrate were infused simultaneously, and ketone body production was calculated from either acetoacetate (AcAc) single-isotope data, beta-hydroxybutyrate (beta-OHB) single-isotope data, the sum of individual fluxes, or the two-pool model. In fasted animals, both the AcAc single-isotope calculation and the sum of individual fluxes overestimated net hepatic production by approximately 50% (P less than 0.05), whereas the beta-OHB single-isotope calculation and the two-pool model gave accurate estimates. In the diabetic animals, the beta-OHB single-isotope calculation underestimated net hepatic production by approximately 30% (P less than 0.05). The sum of individual fluxes overestimated net hepatic production by approximately 46% (P less than 0.05), whereas both the AcAc single-isotope calculation and the two-pool model gave accurate estimates. In conclusion, single-isotope methods give erroneous estimates of net hepatic production of ketone bodies. In contrast, a two-pool model provided an accurate estimate of net hepatic production and thus appears to be suitable for determination of ketone body kinetics in humans

  10. Novel Synthesis and Anti-Microbial Activity Study of Innovative Mannich Bases Containing 2- Phenoxy-1, 3, 2-Dioxa Phospholanes And Indole Systems

    Subba Narayana Kanchana

    2014-09-01

    Full Text Available In the present study, synthesis, characterization, antimicrobial activity on some novel pyrazolone derivatives has been taken up. The primary amines were diazotized with sodium nitrite and HCl mixture at 0-5 0C which were further coupled with ethyl acetoacetic ester to afford phenyl diazonium acetoacetic esters (II. Condensation of 4-substituted aryl hydrazono acetoacetic ester, 4-hydrazinylbenzenamine in the presence of catalytic amount of DMF under microwave irradiation afforded (4Z-4-(2-substituted aryl hydrazono-1-(4-aminophenyl-3-methyl-1H-pyrazol-5(4H-one (III. Subjecting (III to anhydrous K2CO3 in DMF for 8 hours afforded 2-{4-[(4Z-4-(2-substituted aryl hydrazono-4, 5-dihydro-3-methyl-5-oxopyrazol-1-yl] phenyl amino} acetate( IV which was further reacted with hydrazine hydrate to afford 2-{4-[(4Z-4-(2-substituted aryl hydrazono-4, 5-dihydro-3-methyl-5-oxopyrazol-1-yl] phenylamino} acetohydrazide (V. Reaction of (V and Isatin (VI b in DMF afforded (4Z-2-(4-((15Z-4-(2-phenylhydrazono-4,5-dihydro-3-methyl-5 oxopyrazol-1-ylphenylamino-N'-(2-oxoindolin-3-ylideneacetohydrazide (VII. Compound (VII was subsequently subjected to Mannich reaction with cyclic secondary amines (piperidine/morpholine/N-methyl piperidine in the presence of formaldehyde in DMF to afford (4Z-2-(4-((15Z-4-(2-substituted aryl hydrazono-4,5-dihydro-3-methyl-5-oxopyrazol-1-ylphenylamino-N'-(2-oxo-1-((piperidin-1-ylmethylindolin-3-ylideneacetohydrazide (VIII which was characterized by I.R, 1HNMR and Mass data. The anti-bacterial activity of synthesized compounds was studied by the disc diffusion method against pathogenic bacteria and fungi.

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

    MUHAMMED BASHEER UMMATHUR

    2009-03-01

    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.

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

    Cláudia O. Veloso

    2011-01-01

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

    RAHUL P. GAVANDE

    2003-10-01

    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.

  14. Estudos de proteção da célula de saccharomyces cerevisiae para utilização em reações de redução em meio orgânico

    Nascimento Maria da Graça

    2002-01-01

    Full Text Available New methodologies for protection of Saccharomyces cerevisiae (FP cells when supported in montmorillonite K10 (K10, recovered or not with gelatin (G and in the presence or absence of sucrose (S are presented. These systems were used for the enantioselective reduction of ethyl acetoacetate and a-chloroacetophenone in hexane, under FP/K10/G/S and FP/S at 20ºC during 24 hours, affording S-(+-ethyl-3-hydroxybutanoate in 100% conversion and 99% ee, and R-(--2-chloro-1-phenylethanol 79% and 78% ee at 20 and 30 ºC, respectivelly.

  15. 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; Farhad Hatamjafari

    2014-01-01

    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 environ...

  16. 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

    2015-05-01

    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.

  17. The Hantzsch Ester Production in a Water-based Biphasic Medium, Using Infrared Irradiation as the Activating Source

    Raquel Gómez Pliego; Roberto Osnaya; Isai Zamora; Benjamín Velasco Bejarano; Gabriel Arroyo; Eduardo Ramírez San Juan; José Trujillo; Francisco Delgado; René Miranda

    2007-01-01

    The reaction of a wide number of benzaldehydes (1a-n) with ethyl (2a) or methyl (2b) acetoacetate and aqueous ammonia (3) has been carried out in the absence of an organic solvent, and using for the first time infrared irradiation as the heating mode. Thus, under the green chemistry protocole, the corresponding mono-1,4-dihydropyridines (4a-j) and bis-1,4-dihydropyridines (4k-n) were obtained in good yields and short reaction times.

  18. Hyperketonemia increases monocyte adhesion to endothelial cells and is mediated by LFA-1 expression in monocytes and ICAM-1 expression in endothelial cells

    Rains, Justin L.; Jain, Sushil K.

    2011-01-01

    Frequent episodes of hyperketonemia are associated with a higher incidence of vascular disease. The objective of this study was to examine the hypothesis that hyperketonemia increases monocyte-endothelial cell (EC) adhesion and the development of vascular disease in diabetes. Human U937 and THP-1 monocyte cell lines and human umbilical vein endothelial cells (HUVECs) were cultured with acetoacetate (AA) (0–10 mM) or β-hydroxybutyrate (BHB) (0–10 mM) for 24 h prior to evaluating adhesion and a...

  19. Four-component one-pot synthesis of unsymmetrical polyhydroquinoline derivatives using 3-methyl-1-sulfonic acid imidazolium hydrogen sulfate as a catalyst

    Nader Ghaffari Khaligh

    2014-01-01

    3-Methyl-1-sulfonic acid imidazolium hydrogen sulfate has been used as an efficient, halogen-free, and reusable Brönsted acidic ionic liquid catalyst for the synthesis of ethyl-4-aryl/heteryl- hexahy-dro-trimehtyl-5-oxoquinoline-3-carboxylates via the one-pot condensation of dimedone with ar-yl/heteryl aldehydes, ethyl acetoacetate, and ammonium acetate under solvent-free conditions. This method has the advantage of being clean and simple, as well as providing the desired product in high yield over a short reaction time. Furthermore, the catalyst could be recycled and reused four times without any discernible reduction in activity.

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

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

    2004-01-01

    - 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 of the...... Michael adducts. The ionic liquid [bmim][PF 6] can be recovered and repeatedly used in the reactions.......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...

  1. 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

    2015-12-15

    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. PMID:26546212

  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

    2011-09-01

    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. Enzymes involved in 3,5-diaminohexanoate degradation by Brevibacterium sp.

    Barker, H. A.; Kahn, J. M.; Chew, S

    1980-01-01

    Cell-free extracts of Brevibacterium sp. L5 grown on DL-erythro-3,5-diaminohexanoate were found to contain a 3-keto-5-aminohexanoate cleavage enzyme that converts 3-keto-5-aminohexanoate and acetyl-coenzyme A (CokA) to 3-aminobutyryl-CoA and acetoacetate and a deaminase that coverts L-3-aminobutyryl-CoA to crotonyl-CoA. The cleavage enzyme has been purified extensively, and some of its properties have been determined for comparison with the 3-keto-6-acetamido-hexanoate cleavage enzyme of Pseu...

  4. Synthesis of carbon-14 labelled (5Z)-4-bromo-5-(bromomethylene)-2(5H)-furanone:

    Persson, T.; Johansen, S.K.; Martiny, L.;

    2004-01-01

    The potent quorum sensing inhibitor (5Z)-4-bromo-5-(bromomethylene)-2(5H)-[2-C-14]furanone has been prepared in five steps in 7.7% overall yield starting from bromo[1-C-14]acetic acid. Condensation of ethyl bromo[1-C-14]acetate with ethyl acetoacetate followed by decarboxylation was accelerated by...... microwave heating to afford [1-C-14]levulinic acid. Subsequently, bromination and oxidation gave the targeted furan-2-one with a radiochemical purity of > 97% and a specific activity of 57 mCi/mmol....

  5. 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

    2012-09-01

    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.

  6. Activation of Acetone and Other Simple Ketones in Anaerobic Bacteria.

    Heider, Johann; Schühle, Karola; Frey, Jasmin; Schink, Bernhard

    2016-01-01

    Acetone and other ketones are activated for subsequent degradation through carboxylation by many nitrate-reducing, phototrophic, and obligately aerobic bacteria. Acetone carboxylation leads to acetoacetate, which is subsequently activated to a thioester and degraded via thiolysis. Two different types of acetone carboxylases have been described, which require either 2 or 4 ATP equivalents as an energy supply for the carboxylation reaction. Both enzymes appear to combine acetone enolphosphate with carbonic phosphate to form acetoacetate. A similar but more complex enzyme is known to carboxylate the aromatic ketone acetophenone, a metabolic intermediate in anaerobic ethylbenzene metabolism in denitrifying bacteria, with simultaneous hydrolysis of 2 ATP to 2 ADP. Obligately anaerobic sulfate-reducing bacteria activate acetone to a four-carbon compound as well, but via a different process than bicarbonate- or CO2-dependent carboxylation. The present evidence indicates that either carbon monoxide or a formyl residue is used as a cosubstrate, and that the overall ATP expenditure of this pathway is substantially lower than in the known acetone carboxylase reactions. PMID:26958851

  7. High Glass Transition Temperature Renewable Polymers via Biginelli Multicomponent Polymerization.

    Boukis, Andreas C; Llevot, Audrey; Meier, Michael A R

    2016-04-01

    A novel and straightforward one-pot multicomponent polycondensation method was established in this work. The Biginelli reaction is a versatile multicomponent reaction of an aldehyde, a β-ketoester (acetoacetate) and urea, which can all be obtained from renewable resources, yielding diversely substituted 3,4-dihydropyrimidin-2(1H)-ones (DHMPs). In this study, renewable diacetoacetate monomers with different spacer chain lengths (C3, C6, C10, C20) were prepared via simple transesterification of renewable diols and commercial acetoacetates. The diacetoacetate monomers were then reacted with renewable dialdehydes, i.e., terephthalaldehyde and divanillin in a Biginelli type step-growth polymerization. The obtained DHMP polymers (polyDHMPs) displayed high molar masses, high glass transition temperatures (Tg) up to 203 °C and good thermal stability (Td5%) of 280 °C. The Tg of the polyDHMPs could be tuned by variation of the structure of the dialdehyde or the diacetoacetate component. PMID:26800511

  8. Measurement of acetol in serum.

    Casazza, J P; Fu, J L

    1985-08-01

    A method for the derivatization of acetol (1-hydroxyacetone) with 2,4-dinitrophenylhydrazine (DNPH) and for the measurement of the acetol dinitrophenylhydrazone derivative (acetol-DNPH) by high-performance liquid chromatography is presented. The chromatographic separation described here resulted in baseline resolution of the acetol-DNPH peak. Peak integration was proportional to serum acetol concentration over a 5- to 500-nmol/ml range. No other method for the determination of acetol in serum currently exists. Serum from rats in diabetic ketoacidosis was found to contain 11.2 +/- 1.1 nmol acetol/ml serum (N = 3). Serum from a 21-day-fasted human contained 16 nmol/ml acetol. Serum from rats maintained on drinking water containing 1% acetone (v:v) for 6 days contained 152 +/- 31 nmol/ml acetol (N = 5). The presence of acetol in serum under conditions where acetoacetate and acetone are chronically elevated suggests that acetoacetate may be converted to glucose through the conversion of acetone to acetol and L-1,2-propanediol. PMID:3933378

  9. 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

    2008-01-01

    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.

  10. Isolation of a new butanol-producing Clostridium strain: high level of hemicellulosic activity and structure of solventogenesis genes of a new Clostridium saccharobutylicum isolate.

    Berezina, Oksana V; Brandt, Agnieszka; Yarotsky, Sergey; Schwarz, Wolfgang H; Zverlov, Vladimir V

    2009-10-01

    New isolates of solventogenic bacteria exhibited high hemicellulolytic activity. They produced butanol and acetone with high selectivity for butanol (about 80% of butanol from the total solvent yield). Their 16S rDNA sequence was 99% identical to that of Clostridium saccharobutylicum. The genes responsible for the last steps of solventogenesis and encoding crotonase, butyryl-CoA dehydrogenase, electron-transport protein subunits A and B, 3-hydroxybutyryl-CoA dehydrogenase, alcohol dehydrogenase, CoA-transferase (subunits A and B), acetoacetate decarboxylase, and aldehyde dehydrogenase were identified in the new C. saccharobutylicum strain Ox29 and cloned into Escherichia coli. The genes for crotonase, butyryl-CoA dehydrogenase, electron-transport protein subunits A and B, and 3-hydroxybutyryl-CoA dehydrogenase composed the bcs-operon. A monocistronic operon containing the alcohol dehydrogenase gene was located downstream of the bcs-operon. Genes for aldehyde dehydrogenase, CoA-transferase (subunits A and B), and acetoacetate decarboxylase composed the sol-operon. The gene sequences and the gene order within the sol- and bcs-operons of C. saccharobutylicum Ox29 were most similar to those of Clostridium beijerinckii. The activity of some of the bcs-operon genes, expressed in heterologous E. coli, was determined. PMID:19674858

  11. Ketone bodies and islet function: 45Ca handling, insulin synthesis, and release

    D-(-)-beta-Hydroxybutyrate and acetoacetate cause a rapid, sustained, and rapidly reversible stimulation of insulin release from rat pancreatic islets incubated in the presence, but not absence, of D-glucose. This coincides with stimulation of both proinsulin biosynthesis and 45Ca net uptake. The ketone bodies also decrease 45Ca outflow from prelabeled islets perifused in the absence of Ca2+ and, in contrast, enhance effluent radioactivity in the presence of Ca2+. In the presence of D-glucose, the secretory response to D-(-)-beta-hydroxybutyrate is concentration related in the 2.5-20 mM range, abolished in the absence of Ca2+ or presence of KCN, and enhanced by theophylline and forskolin. It corresponds grossly to a shift to the left of the sigmoidal curve relating insulin output to the ambient concentration of D-glucose. The secretory, biosynthetic, and cationic response to acetoacetate is less marked than that evoked by an equimolar concentration of D-(-)-beta-hydroxybutyrate. These features are compatible with the view that the insulinotropic action of ketone bodies would be causally linked to their metabolism in islet cells

  12. Ketone bodies and islet function: sup 45 Ca handling, insulin synthesis, and release

    Malaisse, W.J.; Lebrun, P.; Yaylali, B.; Camara, J.; Valverde, I.; Sener, A. (Brussels Free Univ. (Belgium))

    1990-07-01

    D-(-)-beta-Hydroxybutyrate and acetoacetate cause a rapid, sustained, and rapidly reversible stimulation of insulin release from rat pancreatic islets incubated in the presence, but not absence, of D-glucose. This coincides with stimulation of both proinsulin biosynthesis and {sup 45}Ca net uptake. The ketone bodies also decrease {sup 45}Ca outflow from prelabeled islets perifused in the absence of Ca{sup 2+} and, in contrast, enhance effluent radioactivity in the presence of Ca{sup 2+}. In the presence of D-glucose, the secretory response to D-(-)-beta-hydroxybutyrate is concentration related in the 2.5-20 mM range, abolished in the absence of Ca{sup 2+} or presence of KCN, and enhanced by theophylline and forskolin. It corresponds grossly to a shift to the left of the sigmoidal curve relating insulin output to the ambient concentration of D-glucose. The secretory, biosynthetic, and cationic response to acetoacetate is less marked than that evoked by an equimolar concentration of D-(-)-beta-hydroxybutyrate. These features are compatible with the view that the insulinotropic action of ketone bodies would be causally linked to their metabolism in islet cells.

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

    Hijaz, Faraj; Killiny, Nabil

    2016-05-20

    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. PMID:27106399

  14. Preparation of toluenes specifically labelled with C-14

    The attempt to prepare toluene-2 C-14 starting from ethyl acetoacetate failed because the reaction of sodium ethyl acetoacetate with dibromopropane could not be performed. This labelling was also attempted starting from methyl tetrahydropyran. This synthesis was not successful because the preparationof 2-methyl pimelic acid was impossible. Labellimg of carbon-3 of toluene starting from succinic acid could not be carried out because the Grignard reaction of a-ketoglutaric acid as unsuccessful. The radioactive syntheses started from propanediol resp. from c-butyrolactone. (The labelling of carbon-3 of toluene was not possible in this case because the reduction of malonic acid to propanediol could not be carried out). The synthesis course was identical for all labellings. The synthesis differed only in the introduction of the activiy from potassium cyanide C-14. The starting compounds were reacted to propanedinitril, which was saponified to glutaric acid. The acid was reduced to pentanediol. From the diol, dibromopentane was produced. Reaction with potassium cyanide yielded, after saponification, pimelic acid. The acid was cyclizated to cyclohexanone, which was reduced to cyclohexanol. The OH group was replaced by bromine and subsequently by a methyl group. Dehydration yielded toluene. The following labellings were carried out: Toluene-2, -2.6, -1.2.6, -1.2 C-14. (G.G.)

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

    José Clecildo Barreto Bezerra

    1999-11-01

    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.

  16. 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

    2016-01-01

    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. PMID:26476882

  17. Effects of accumulated 60Co γ-ray irradiation on the small molecular metabolites in rats urine

    Objective: To explore the effects of accumulated 60Co γ-ray irradiation on small molecular metabolites in rats urine. Methods: Ten healthy male SD rats were irradiated by 60Co γ-rays in 5 days and the accumulated doses were 0, 0.2, 0.4, 0.6, 0.8, 1.0 Gy, respectively. The metabolites in urine of different groups were measured with 1H-NMR combined with principal components analysis (PCA) and partial least square discriminate analysis (PLS-DA). Results: The metabolites in rat urine were obviously changed after irradiation. Compared with control group, the amount of acetoacetate decreased after irradiation (t=29.7-30.7, P<0.05), but its relative level was stable when the dose increased (P>0.05). Meanwhile,the relative level of hippuric acid increased (t=4.4-21.6, P<0.05) especially when the accumulated dose was higher than 1 Gy (t=21.6, P<0.05). The relative level of proline, taurine and trimethylamine-N-oxide increased after irradiation with the same trend(t=3.5-13.4, 4.7-11.5, 2.9-12.7, P<0.05). Conclusions: The acetoacetate, hippuric acid, proline, taurine, and trimethylamine-N-oxide may be applicable for biomarkers of accumulative irradiation on rat. (authors)

  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

    2015-08-01

    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. PMID:26145173

  19. Measuring changes in substrate utilization in the myocardium in response to fasting using hyperpolarized [1-(13)C]butyrate and [1-(13)C]pyruvate.

    Bastiaansen, Jessica A M; Merritt, Matthew E; Comment, Arnaud

    2016-01-01

    Cardiac dysfunction is often associated with a shift in substrate preference for ATP production. Hyperpolarized (HP) (13)C magnetic resonance spectroscopy (MRS) has the unique ability to detect real-time metabolic changes in vivo due to its high sensitivity and specificity. Here a protocol using HP [1-(13)C]pyruvate and [1-(13)C]butyrate is used to measure carbohydrate versus fatty acid metabolism in vivo. Metabolic changes in fed and fasted Sprague Dawley rats (n = 36) were studied at 9.4 T after tail vein injections. Pyruvate and butyrate competed for acetyl-CoA production, as evidenced by significant changes in [(13)C]bicarbonate (-48%), [1-(13)C]acetylcarnitine (+113%), and [5-(13)C]glutamate (-63%), following fasting. Butyrate uptake was unaffected by fasting, as indicated by [1-(13)C]butyrylcarnitine. Mitochondrial pseudoketogenesis facilitated the labeling of the ketone bodies [1-(13)C]acetoacetate and [1-(13)C]β-hydroxybutyryate, without evidence of true ketogenesis. HP [1-(13)C]acetoacetate was increased in fasting (250%) but decreased during pyruvate co-injection (-82%). Combining HP (13)C technology and co-administration of separate imaging agents enables noninvasive and simultaneous monitoring of both fatty acid and carbohydrate oxidation. This protocol illustrates a novel method for assessing metabolic flux through different enzymatic pathways simultaneously and enables mechanistic studies of the changing myocardial energetics often associated with disease. PMID:27150735

  20. Synthesis, spectral, catalytic and antimicrobial studies of PPh 3/AsPh 3 complexes of Ru(II) with dibasic tridentate O, N, S donor ligands

    Balasubramanian, K. P.; Karvembu, R.; Prabhakaran, R.; Chinnusamy, V.; Natarajan, K.

    2007-09-01

    Complexes of the type [Ru(CO)(EPh 3)(B)(L)] (E = P or As; B = PPh 3, AsPh 3, py or pip; L = dianion of the Schiff bases derived from thiosemicarbazone with acetoacetanilide, acetoacet- o-toluidide and o-chloro acetoacetanilide) have been synthesized from the reactions of equimolar amounts of [RuHCl(CO)(EPh 3) 2(B)] and Schiff bases in benzene. The new complexes have been characterized by analytical and spectral (IR, electronic, NMR) data. The arrangement of PPh 3 groups around ruthenium metal was determined from 31P NMR spectra. An octahedral structure has been assigned for all the new complexes. All the complexes exhibited catalytic activity for the oxidation of benzyl alcohol and cyclohexanol in presence of N-methylmorpholine- N-oxide as co-oxidant. The complexes also exhibited antibacterial activity against E. coli, Aeromonas hydrophilla and Salmonella typhi. The activity was compared with standard streptomycin.

  1. Green Synthesis, Characterization and Electrochemical Behavior of New Thiazole Based Coumarinyl Scaffolds.

    Saeed, Aamer; Channar, Pervaiz Ali; Shabir, Ghulam; Larik, Fayaz Ali

    2016-05-01

    The present paper deals with the synthesis of thiazolo-coumarin derivatives, prepared starting from naphthaldehyde, ethyl acetoacetate and hydrazine in three steps and characterized by spectroscopic analysis. UV-Visible spectra of the compounds was carried out in different solvents DMF, ethanol, methanol, ethyl acetate and acetone and the absorption was observed in the range 338-390 nm. Electrochemical study of thiazoles was conducted in DMF and redox behavior was examined. Fluorescence carried out in ethanol showed sharp emission in the range 440-505 nm. Graphical Abstract Synthesis, characterization and electrochemical behavior of new thiazole based coumarinyl Scaffolds. Aamer Seed, Pervaiz Ali Channar, Ghulam Shabir, Fayaz Ahmed Larik. PMID:27067209

  2. 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

    2016-01-01

    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. PMID:26907242

  3. "Synthesis and smooth muscle Calcium channel antagonist effects of new derivatives of 1,4-Dihydropyridine containing Nitroimidazol substituent "

    Miri R

    2002-09-01

    Full Text Available A group of racemic 3-[(2-hydroxyethyl, (2-Methoxyethyl, (2-acetylethyl or (2-cyanoethyl], 5- methyl, ethyl or isopropyl-1, 4-dihydro-2, 6-dimethyl-4-(1-methyl-5-nitro-2-imidazolyl-3, 5-pyridinedicarboxylates [XIV-XXV] were prepared by the reaction of 1-methyl-5-nitroimidazol-2-carboxaldehyde [X] with acetoacetic esters [VI-IX] and alkys 3-aminocrotonate [XI-XIII]. In vitro calcium channel antagonist activities of the tested compounds were determined by their effects on contraction of Guinea Pig Ileal Longitudinal Smooth Muscle (GPILSM which was induced by carbacol (1.67 χ 10^-7 M. All compounds exhibited calcium channel antagonist activity (IC50=10^-12 to 10^-13 M range comparable to nifedipine as reference drug (IC50=1.07±0.12x 10^-11 M.

  4. Photochemical and hot H atom reactions of acetaldehyde

    Photolysis and hot hydrogen atom reactions of acetaldehyde have been studied simultaneously not only with a cadmium but also with a zinc resonance lamp as the light source. Contrary to previous results, new products (acetone, ethanol, 2-propanol, ethyl acetoacetate, and diacetone alcohol) are identified in the photolysis of acetaldehyde. On the other hand, the principal products in decreasing order from the hot atom reaction with acetaldehyde have been established to be hydrogen, biacetyl, methane, carbon monoxide, and ethanol, whereas acetone and ethylene existed as minor products. A study of product quantum yields at 214 nm vs. various [CH3CHO]/[CH2S] ratios are reported. The formation of several unexpected products in these studies strongly suggested that the aldehyde functional group of acetaldehyde has an unsaturated character relative to the addition of both hydrogen atoms and methyl radicals. This latter new observation and accompanying mechanisms are discussed. 2 figures, 6 tables

  5. 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

    2014-08-01

    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.

  6. 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: samieh.fozooni@uk.ac.ir, E-mail: s_fozooni@yahoo.com [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

    2013-10-15

    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)

  7. SYNTHESIS, CROSSLINKING MECHANISM AND PROPERTIES OF A POLYACRYLATE/POLYURETHANE COMPOSITE COATING

    TANG Liming; GUO Wei; ZHOU Qixiang

    1997-01-01

    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.

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

    Khodabakhsh Niknam; Nassim Borazjani; Reza Rashidian; Abbas Jamali

    2013-01-01

    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.

  9. Reassessment of 14CO2 compartmentation and of [14C]formate oxidation in rat liver

    Our previous report had concluded that a fraction of [14C]formate oxidation in liver occurs in the mitochondrion. This conclusion was based on the labeling patterns of urea and acetoacetate labeled via 14CO2 generated from [14C]formate and other [14C]substrates. We reassessed our interpretation in experiments conducted in (i) perifused mitochondria and (ii) isolated livers perfused with buffer containing [14C]formate, [14C]gluconolactone, 14CO2, or NaH13CO3, in the absence and presence of acetazolamide, an inhibitor of carbonic anhydrase. Our data show that the cytosolic pools of bicarbonate and CO2 are not in isotopic equilibrium when 14CO2 is generated in the cytosol or is supplied as NaH14CO3. We retract our earlier suggestion of a mitochondrial site of [14C]formate oxidation

  10. 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

    2015-09-01

    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.

  11. Ultrasound Irradiation Effects on the Michael Reaction Catalyzed by KF/basic Al2O3%超声波辐射下KF/basic Al2O3催化的Michael加成

    李记太; 陈国锋; 李同双

    2002-01-01

    超声辐射下,对查尔酮与含活泼亚甲基的化合物诸如丙二酸二乙酯、硝基甲烷、乙酰乙酸乙酯、环己酮等,在KF/basic Al2O3催化下的Michael 反应进行了研究,该法具有反应时间短、产率高、操作简单、催化剂可以重复使用的优点.%The Michael reaction of chalcones as acceptors with several active methylene compounds such as diethyl malonate, nitromethane,ethyl acetoacetate and cyclohexone et al as donors catalyzed by KF/basic Al2O3 under ultrasound irradiation proceed very efficiently and furnish higher yield of adducts.

  12. A green four-component synthesis of zwitterionic alkyl/benzyl pyrazolyl barbiturates and their photophysical studies.

    Bihani, Manisha; Bora, Pranjal P; Bez, Ghanashyam; Askari, Hassan

    2014-11-01

    A novel series of unsymmetrically substituted alkyl/benzyl pyrazolyl barbiturates incorporating highly biologically active pyrazolone and barbiturate moieties was synthesized by four-component reactions of a mixture of ethyl acetoacetate, hydrazine hydrate, aldehydes and barbituric acid/thiobarbituric acid in ethanol without using a catalyst. The photophysical properties of the newly designed alkyl/benzyl pyrazolyl barbiturates were studied, and good quantum yield of some products indicated a definitive scope in the field of biochemical applications. Single-crystal X-ray crystallographic studies revealed that the newly synthesized compounds exist in zwitterionic form. The zwitterionic nature of the new chimera makes them interesting candidates for drug delivery as zwitterionic drugs are known to have highly water soluble properties, specific protein absorption, slow recognition by immune system, slow blood clearance from body and can constantly diffuse and deposit throughout the physiological pH. PMID:25005075

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

    Rishiram Prajuli

    2015-12-01

    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.

  14. Synthesis of new 2-amino-4H-pyran-3,5-dicarboxylate derivatives using nanocrystalline MIIZr4(PO4)6 ceramics as reusable and robust catalysts under microwave irradiation

    In the present paper, we report the successful synthesis of nanocrystalline MIIZr4(PO4)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-MIIZr4(PO4)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 MnZr4(PO4)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. 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

    2016-01-01

    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. PMID:27563914

  16. Synthesis of Some Novel Biologically Active Disperse Dyes Derived from 4-Methyl-2,6-dioxo-1-propyl-1,2,5,6-tetrahydro-pyridine-3-carbonitrile as Coupling Component and Their Colour Assessment on Polyester Fabrics

    Mohamed H. Elnagdi

    2012-07-01

    Full Text Available A series of novel azo-disperse dyes containing alkylhydrazonopyridinone structures were synthesized. 4-Methyl-2,6-dioxo-1-propyl-1,2,5,6-tetrahydropyridine-3-carbonitrile (8 is synthesized by one-pot synthesis using ethyl cyanoacetate, propylamine, and ethyl acetoacetate. Compound 8 is then coupled with aromatic and heteroaromatic diazonium salts to afford the corresponding aryl- and heteroaryl-4-methyl-2,6-dioxo-1-propyl-1,2,5,6-tetrahydropyridine-3-carbonitriles 12a,b and 13a–c. Structural assignments to the dyes were made using NMR spectroscopic methods. A high temperature dyeing method was employed to apply these dyes to polyester fabrics. Most of the dyed fabrics tested displayed very good light fastness levels and good wash fastness. Finally, the biological activity of the prepared dyes against Gram positive bacteria and Gram negative bacteria were evaluated.

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

    Rodrigo González-Olvera

    2016-02-01

    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.

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

    Sudipta Pathak

    2013-11-01

    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.

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

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

    2015-04-01

    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).

  20. Metabolomic analysis of amino acid and energy metabolism in rats supplemented with chlorogenic acid

    Ruan, Zheng; Yang, Yuhui; Zhou, Yan; Wen, Yanmei; Ding, Sheng; Liu, Gang; Wu, Xin; Deng, Zeyuan; Assaad, Houssein; Wu, Guoyao

    2016-01-01

    This study was conducted to investigate effects of chlorogenic acid (CGA) supplementation on serum and hepatic metabolomes in rats. Rats received daily intragastric administration of either CGA (60 mg/kg body weight) or distilled water (control) for 4 weeks. Growth performance, serum biochemical profiles, and hepatic morphology were measured. Additionally, serum and liver tissue extracts were analyzed for metabolomes by high-resolution 1H nuclear magnetic resonance-based metabolomics and multivariate statistics. CGA did not affect rat growth performance, serum biochemical profiles, or hepatic morphology. However, supplementation with CGA decreased serum concentrations of lactate, pyruvate, succinate, citrate, β-hydroxybutyrate and acetoacetate, while increasing serum concentrations of glycine and hepatic concentrations of glutathione. These results suggest that CGA supplementation results in perturbation of energy and amino acid metabolism in rats. We suggest that glycine and glutathione in serum may be useful biomarkers for biological properties of CGA on nitrogen metabolism in vivo. PMID:24927697

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

    Meyrambek Ospanov

    2014-10-01

    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.

  2. Isolation and characterization of a new gram-negative, acetone-degrading, nitrate-reducing bacterium from soil, Paracoccus solventivorans sp. nov.

    Siller, H; Rainey, F A; Stackebrandt, E; Winter, J

    1996-10-01

    An acetone-degrading, nitrate-reducing, coccoid to rod-shaped bacterium, strain L1, was isolated from soil on the site of a natural gas company. Cells of the logarithmic growth phase reacted gram positive, while those of the stationary growth phase were gram negative. Single organisms were 0.4 to 0.5 by 0.9 to 1.5 microns in size, nonmotile, and non-spore forming and had poly-beta-hydroxybutyrate inclusions. The doubling time of strain L1 on acetone-CO2-nitrate at the optimal pH of 7 to 8 and the optimal temperature of 30 to 37 degrees C was 12 h. More than 0.2% NaCl or 10 mM thiosulfate inhibited growth. For oxygen or nitrate respiration, acetone and a few organic chemicals were utilized as carbon sources whereas many others could not be used (for details, see Results). Bicarbonate (or CO2) was essential for growth on acetone but not for growth on acetoacetate. The growth yields for acetone-CO2 and acetoacetate were 28.3 and 27.3 g/mol, respectively. With acetone as the carbon source, poly-beta-hydroxybutyrate accounted for up to 40% of the cellular dry weight. The DNA of strain L1 had a G + C content of 68.5 mol% (as determined by high-performance liquid chromatography of nucleotides) or 70 mol% (as determined by the TM method). The sequence of the gene coding for the 16S rRNA led to the classification of strain L1 in the paracoccus group of the alpha subclass of the Proteobacteria. The new isolate is named Paracoccus solventivorans sp. nov. DSM 6637. PMID:8863446

  3. Protein and amino acid metabolism in skeletal muscle

    Wu, Guoyao.

    1989-01-01

    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.

  4. Usefulness of postmortem biochemistry in identification of ketosis: Diagnosis of ketoacidosis at the onset of autoimmune type 1 diabetes in an autopsy case with cold exposure and malnutrition.

    Tani, Naoto; Michiue, Tomomi; Chen, Jian-Hua; Oritani, Shigeki; Ishikawa, Takaki

    2016-09-01

    A severely malnourished, Japanese female in her twenties was found dead in her apartment. On autopsy, most of the findings from the internal examination were suggestive of hypothermia. Postmortem biochemistry, however, showed severely increased levels of glycated hemoglobin (HbA1c) and blood and urine glucose levels. Levels of acetone, 3-hydroxybutyric acid, and acetoacetate in various body fluids were also highly increased, indicating ketosis. The serum insulin and c-peptide levels were severely low, and subsequent testing was positive for anti-GAD antibodies. Immunohistochemical examination of the pancreatic islet cells revealed few insulin-positive cells but many glucagon-positive cells on staining. Furthermore, slight invasion of CD8-positive lymphocytes in the pancreatic islets of Langerhans was observed. Results of immunostaining of the pancreatic and bronchial epithelial tissues were partly positive for the Influenza A virus. We concluded that severe ketoacidosis associated with rapid-onset hyperglycemia due to autoimmune type 1 diabetes (AT1D) had occurred shortly before death. However, the ketosis was accompanied by hypothermia and malnutrition as well as diabetic ketoacidosis (DKA). Therefore, we retrospectively collected biochemical data on cases of hypothermia and malnutrition and compared them with the present case. Serum glucose, acetone, 3-hydroxybutyric acid, and acetoacetic acid can be used for screening and diagnosis to distinguish DKA from ketosis due to hypothermia and malnutrition. Therefore, in the present case, we diagnosed that the natural cause of death was due to AT1D. In conclusion, screening investigations for relevant biochemical markers can provide essential information for the diagnosis of metabolic disturbances, which fail to demonstrate characteristic autopsy findings. PMID:27591535

  5. Structural Insights into an Oxalate-producing Serine Hydrolase with an Unusual Oxyanion Hole and Additional Lyase Activity.

    Oh, Juntaek; Hwang, Ingyu; Rhee, Sangkee

    2016-07-15

    In Burkholderia species, the production of oxalate, an acidic molecule, is a key event for bacterial growth in the stationary phase. Oxalate plays a central role in maintaining environmental pH, which counteracts inevitable population-collapsing alkaline toxicity in amino acid-based culture medium. In the phytopathogen Burkholderia glumae, two enzymes are responsible for oxalate production. First, the enzyme oxalate biosynthetic component A (ObcA) catalyzes the formation of a tetrahedral C6-CoA adduct from the substrates acetyl-CoA and oxaloacetate. Then the ObcB enzyme liberates three products from the C6-CoA adduct: oxalate, acetoacetate, and CoA. Interestingly, these two stepwise reactions are catalyzed by a single bifunctional enzyme, Obc1, from Burkholderia thailandensis and Burkholderia pseudomallei Obc1 has an ObcA-like N-terminal domain and shows ObcB activity in its C-terminal domain despite no sequence homology with ObcB. We report the crystal structure of Obc1 in its apo and glycerol-bound form at 2.5 Å and 2.8 Å resolution, respectively. The Obc1 N-terminal domain is essentially identical both in structure and function to that of ObcA. Its C-terminal domain has an α/β hydrolase fold that has a catalytic triad for oxalate production and a novel oxyanion hole distinct from the canonical HGGG motif in other α/β hydrolases. Functional analyses through mutagenesis studies suggested that His-934 is an additional catalytic acid/base for its lyase activity and liberates two additional products, acetoacetate and CoA. These results provide structural and functional insights into bacterial oxalogenesis and an example of divergent evolution of the α/β hydrolase fold, which has both hydrolase and lyase activity. PMID:27226606

  6. 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

    2016-04-01

    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. PMID:26969155

  7. 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.

  8. [The different notions about beta-oxidation of fatty acids in peroxisomes, peroxisomes and ketonic bodies. The diabetic, acidotic coma as an acute deficiency of acetyl-CoA and ATP].

    Kotkina, T I; Titov, V N; Parkhimovich, R M

    2014-03-01

    The mechanisms of beta-oxidation of fatty acids developed more than a century before have no compliance with actual physical chemical data. The oxidation of long-chain C 16:0 palmitic saturated fatty acid occurs not by sequential formation of eight molecules of acetyl-KoA but by force of formation of double bond and its hydrolysis on two short-chain C 8:0 fatty acids. Only short-chain fatty acids can become shorter under "chipping" of C 2-acetate with formation of C 4-butyric acid (butyrate) and its metabolites (beta-hidroxibutirate, acetoacetate, acetone). The critical moment of oxidation is a hydrolysis of acetoacetyl-KoA on two molecules of acetyl-KoA. The molecule of ATP is to be expended on hydrolysis. The foundation of nonspecific biological reaction of stress--ketoacidosis,--is a decrease in mitochondrions of acetyl-KoA pool formed both from glycogen and glucose and fatty acids. The oxalate acetate inputs into Krebs cycle inadequate amount of acetyl-KoA which limits synthesis of ATP. The insulin has no direct involvement into development of ketoacidosis but prepares conditions to facilitate nonspecific etiological factor to initiate diabetic ketoacidosis. These are the pooling of small amount of glycogen in cytozol and the predominance in cytozol of cells and adipocytes of palmitic triglycerides which are slowly hydrolyzed by hormone-dependent lipase to release non-esterified fatty acids into intercellular medium. The increase of their concentration in blood plasma precedes ketoacidosis which is developing in patients without diabetes mellitus too. When cells begin to oxidize unsaturated linoleic and linolenic acids with large number of double binds instead of medium-chain fatty acids, oleinic and palmitic fatty acids to support beta-oxidation in mitochondrions and synthesis of ATP the amount of butyric acid, beta-hidroxibutiryl-KoA and acetoacetyl-KoA increases and of acetyl-KoA decreases. The cause of fatal outcome is the development of metabolic acidosis

  9. Paper-based microfluidic devices for analysis of clinically relevant analytes present in urine and saliva.

    Klasner, Scott A; Price, Alexander K; Hoeman, Kurt W; Wilson, Rashaun S; Bell, Kayla J; Culbertson, Christopher T

    2010-07-01

    We report the use of paper-based microfluidic devices fabricated from a novel polymer blend for the monitoring of urinary ketones, glucose, and salivary nitrite. Paper-based devices were fabricated via photolithography in less than 3 min and were immediately ready for use for these diagnostically relevant assays. Patterned channels on filter paper as small as 90 microm wide with barriers as narrow as 250 microm could be reliably patterned to permit and block fluid wicking, respectively. Colorimetric assays for ketones and nitrite were adapted from the dipstick format to this paper microfluidic chip for the quantification of acetoacetate in artificial urine, as well as nitrite in artificial saliva. Glucose assays were based on those previously demonstrated (Martinez et al., Angew Chem Int Ed 8:1318-1320, 1; Martinez et al., Anal Chem 10:3699-3707, 2; Martinez et al., Proc Nat Acad Sci USA 50:19606-19611, 3; Lu et al., Electrophoresis 9:1497-1500, 4; Abe et al., Anal Chem 18:6928-6934, 5). Reagents were spotted on the detection pad of the paper device and allowed to dry prior to spotting of samples. The ketone test was a two-step reaction requiring a derivitization step between the sample spotting pad and the detection pad, thus for the first time, confirming the ability of these paper devices to perform online multi-step chemical reactions. Following the spotting of the reagents and sample solution onto the paper device and subsequent drying, color images of the paper chips were recorded using a flatbed scanner, and images were converted to CMYK format in Adobe Photoshop CS4 where the intensity of the color change was quantified using the same software. The limit of detection (LOD) for acetoacetate in artificial urine was 0.5 mM, while the LOD for salivary nitrite was 5 microM, placing both of these analytes within the clinically relevant range for these assays. Calibration curves for urinary ketone (5 to 16 mM) and salivary nitrite (5 to 2,000 microM) were

  10. 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

    2012-09-01

    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.

  11. Graphene quantum dots from graphite by liquid exfoliation showing excitation-independent emission, fluorescence upconversion and delayed fluorescence.

    Sarkar, Suprabhat; Gandla, Dayakar; Venkatesh, Yeduru; Bangal, Prakriti Ranjan; Ghosh, Sutapa; Yang, Yang; Misra, Sunil

    2016-08-21

    Facile synthesis of 2-10 nm-sized graphene quantum dots (GQDs) from graphite powder by organic solvent-assisted liquid exfoliation using a sonochemical method is reported in this study. Synthesized GQDs are well dispersed in organic solvents like ethyl acetoacetate (EAA), dimethyl formamide (DMF) and also in water. MALDI-TOF mass spectrometry reveals its selective mass fragmentation. Detailed characterizations by various techniques like X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), X-ray photoelectron spectroscopy (XPS), Raman spectroscopy and high resolution transmission electron microscopy (HRTEM) confirm the formation of disordered, functional GQDs. Density functional theory (DFT) calculation confirms HOMO-LUMO energy gap variation with changing size and functionalities. Photoluminescence (PL) properties of as-prepared GQDs were studied in detail. The ensemble studies of GQDs showed excellent photoluminescence properties comprising normal and upconverted fluorescence, delayed fluorescence and room-temperature phosphorescence. PL decay dynamics of GQDs has been explored using time-correlated single-photon technique (TCSPC) as well as femtosecond fluorescence upconversion technique. In vitro cytotoxicity study reveals its biocompatibility and high cell viability (>91%) even at high concentration (400 μg mL(-1)) of GQDs in Chinese Hamster Ovary (CHO) cells. PMID:27302411

  12. Natural spectroscopic hydrogen isotope transfer in alcohol dehydrogenase-catalysed reduction

    The enantiomeric purity of natural α-mono deuterated enantiomers, (R) and (S)ethanol-1-d1, in the alcohol produced by sugar fermentation with yeast was studied by 2H NMR using their esters derived from optical mandelic acid. The results of isotope tracing experiments show that the transfer pathways of the two eantiotopic hydrogens of the methylene group are different. It was observed that (S)-deuterium comes only from the medium water. The (R)-deuterium transferred by NADH in alcohol dehydrogenase reduction of the acetaldehyde is complex origin. Some of them originates from carbon bound hydrogen of the sugar, especially from C(4) position of glucose and most of them comes from water. Only a small portion of the NADH deuterium is incorporated indirectly from water through enzyme catalysed exchange between the pro-S site of NADH and flavin. When a carbonyl compound (ethyl acetoacetate) was reduced under the same conditions during the alcoholic fermentation, among the NADH-transferred deuterium, only a small portion comes from water while most comes from the unexchangeable positions of the glucose. (author)

  13. Ketones block amyloid entry and improve cognition in an Alzheimer's model.

    Yin, Jun Xiang; Maalouf, Marwan; Han, Pengcheng; Zhao, Minglei; Gao, Ming; Dharshaun, Turner; Ryan, Christopher; Whitelegge, Julian; Wu, Jie; Eisenberg, David; Reiman, Eric M; Schweizer, Felix E; Shi, Jiong

    2016-03-01

    Sporadic Alzheimer's disease (AD) is responsible for 60%-80% of dementia cases, and the most opportune time for preventive intervention is in the earliest stage of its preclinical phase. As traditional mitochondrial energy substrates, ketone bodies (ketones, for short), beta-hydroxybutyrate, and acetoacetate, have been reported to provide symptomatic improvement and disease-modifying activity in epilepsy and neurodegenerative disorders. Recently, ketones are thought as more than just metabolites and also as endogenous factors protecting against AD. In this study, we discovered a novel neuroprotective mechanism of ketones in which they blocked amyloid-β 42, a pathologic hallmark protein of AD, entry into neurons. The suppression of intracellular amyloid-β 42 accumulation rescued mitochondrial complex I activity, reduced oxidative stress, and improved synaptic plasticity. Most importantly, we show that peripheral administration of ketones significantly reduced amyloid burden and greatly improved learning and memory ability in a symptomatic mouse model of AD. These observations provide us insights to understand and to establish a novel therapeutic use of ketones in AD prevention. PMID:26923399

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

    A.V. Faria

    2011-02-01

    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.

  15. Stereoselective Reduction of Prochiral Ketones by Plant and Microbial Biocatalysts.

    Javidnia, K; Faghih-Mirzaei, E; Miri, R; Attarroshan, M; Zomorodian, K

    2016-01-01

    Chiral alcohols are the key chiral building blocks to many enantiomerically pure pharmaceuticals. The biocatalytic approach in asymmetric reduction of corresponding prochiral ketones to the preparation of these optically pure substances is one of the most promising routes. The stereoselective reduction of different kinds of prochiral ketones catalyzed by various plants and microorganisms was studied in this work. Benzyl acetoacetate, methyl 3-oxopentanoate, ethyl 3-oxopentanoate, and ethyl butyryl acetate were chosen as the model substrates for β-ketoesters. Benzoyl acetonitrile, 3-chloro propiophenone, and 1-acetyl naphthalene were chosen as aromatic aliphatic ketones. Finally, 2-methyl benzophenone and 4-chloro benzophenone were selected as diaryl ketones. Plant catalysis was conducted by Daucus carota, Brassica rapa, Brassica oleracea, Pastinaca sativa, and Raphnus sativus. For microbial catalysis, Aspergillus foetidus, Penicillum citrinum, Saccharomyces carlbergensis, Pichia fermentans, and Rhodotrula glutinis were chosen. Chiral alcohols were obtained in high yields and with optical purity. A superiority in the microorganisms' performance in the bioreduction of prochiral ketones was detected. Among microorganisms, Rhodotrula glutinis showed remarkable results with nearly all substrates and is proposed for future studies. PMID:27168684

  16. Stereoselective reduction of prochiral ketones by plant and microbial biocatalysts

    K Javidnia

    2016-01-01

    Full Text Available Chiral alcohols are the key chiral building blocks to many enantiomerically pure pharmaceuticals. The biocatalytic approach in asymmetric reduction of corresponding prochiral ketones to the preparation of these optically pure substances is one of the most promising routes. The stereoselective reduction of different kinds of prochiral ketones catalyzed by various plants and microorganisms was studied in this work. Benzyl acetoacetate, methyl 3-oxopentanoate, ethyl 3-oxopentanoate, and ethyl butyryl acetate were chosen as the model substrates for β-ketoesters. Benzoyl acetonitrile, 3-chloro propiophenone, and 1-acetyl naphthalene were chosen as aromatic aliphatic ketones. Finally, 2-methyl benzophenone and 4-chloro benzophenone were selected as diaryl ketones. Plant catalysis was conducted by Daucus carota, Brassica rapa, Brassica oleracea, Pastinaca sativa, and Raphnus sativus.For microbial catalysis, Aspergillus foetidus, Penicillum citrinum, Saccharomyces carlbergensis, Pichia fermentans, and Rhodotrula glutinis were chosen. Chiral alcohols were obtained in high yields and with optical purity. A superiority in the microorganisms' performance in the bioreduction of prochiral ketones was detected. Among microorganisms, Rhodotrula glutinis showed remarkable results with nearly all substrates and is proposed for future studies.

  17. Synthesis of labeled 5-hydroxy-6-methyl(4,6-14C)uracil

    β-Uramino(1,3-14C)crotonic ester is prepared for the first time by condensation of urea with labeled (2,4-14C)acetoacetic ester in the presence of ethanol and concentrated HCl. Treatment of β-uramino(1,3-14C)crotonic ester with base gives the Na salt of β-uramino(1,3-14C)crotonic acid. Hydrolysis of the above acid by HCl at 65 degrees gives 6-methyl(4,6-14C)uracil. The oxidation of 6-methyl(4,6-14C)uracil by solid ammonium persulfate in basic solution at 40-70 degrees C gives the intermediate 6-methyl(4,6-14C)uracil-5-ammonium sulfate in 35% yield with 99-100% purity. Hydrolysis of pure 6-methyl(4,6-14C)uracil-5-ammonium sulfate by H2SO4 at 85-95 degrees C gives 5-hydroxy-6-methyl(4,6-14C)uracil in 95-100% yield with 99-100% purity. The product purity is proved by TLC, melting point, elemental analysis, and IR, UV, and PMR spectra

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

    Maeng, Wan Young; Yoo, Mi

    2015-11-01

    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. PMID:26726529

  19. Hyperketonemia and ketosis increase the risk of complications in type 1 diabetes.

    Kanikarla-Marie, Preeti; Jain, Sushil K

    2016-06-01

    Diets that boost ketone production are increasingly used for treating several neurological disorders. Elevation in ketones in most cases is considered favorable, as they provide energy and are efficient in fueling the body's energy needs. Despite all the benefits from ketones, the above normal elevation in the concentration of ketones in the circulation tend to illicit various pathological complications by activating injurious pathways leading to cellular damage. Recent literature demonstrates a plausible link between elevated levels of circulating ketones and oxidative stress, linking hyperketonemia to innumerable morbid conditions. Ketone bodies are produced by the oxidation of fatty acids in the liver as a source of alternative energy that generally occurs in glucose limiting conditions. Regulation of ketogenesis and ketolysis plays an important role in dictating ketone concentrations in the blood. Hyperketonemia is a condition with elevated blood levels of acetoacetate, 3-β-hydroxybutyrate, and acetone. Several physiological and pathological triggers, such as fasting, ketogenic diet, and diabetes cause an accumulation and elevation of circulating ketones. Complications of the brain, kidney, liver, and microvasculature were found to be elevated in diabetic patients who had elevated ketones compared to those diabetics with normal ketone levels. This review summarizes the mechanisms by which hyperketonemia and ketoacidosis cause an increase in redox imbalance and thereby increase the risk of morbidity and mortality in patients. PMID:27036365

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

    Fiskum, G; Lehninger, A L

    1979-07-25

    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. PMID:36390

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

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

    1978-04-01

    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. PMID:25436

  2. Production of chiral alcohols from prochiral ketones by microalgal photo-biocatalytic asymmetric reduction reaction.

    Yang, Zhong-Hua; Luo, Li; Chang, Xu; Zhou, Wei; Chen, Geng-Hua; Zhao, Yan; Wang, Ya-Jun

    2012-06-01

    Microalgal photo-biocatalysis is a green technique for asymmetric synthesis. Asymmetric reduction of nonnatural prochiral ketones to produce chiral alcohols by microalgal photo-biocatalysis was studied in this work. Acetophenone (ACP) and ethyl acetoacetate (EAA) were chosen as model substrates for aromatic ketones and β-ketoesters, respectively. Two prokaryotic cyanophyta and two eukaryotic chlorophyta were selected as photo-biocatalysts. The results proved that nonnatural prochiral ketones can be reduced by microalgal photo-biocatalysis with high enantioselectivity. Illumination is indispensable to the photo-biocatalysis. For aromatic ketone, cyanophyta are eligible biocatalysts. For ACP asymmetric reduction reaction, about 45% yield and 97% e.e. can be achieved by the photo-biocatalysis reaction with Spirulina platensis as biocatalyst. On the contrary, chlorophyta are efficient biocatalysts for β-ketoester asymmetric reduction reaction among the four tested algae. For EAA asymmetric reduction reaction, about 70% yield and 90% e.e. can be achieved with Scenedesmus obliquus as biocatalyst. The microalgae used in this study outperformed other characterized biocatalysts such as microbial and plant cells. PMID:22322691

  3. Metabonomic profiling of renal cell carcinoma: High-resolution proton nuclear magnetic resonance spectroscopy of human serum with multivariate data analysis

    Metabonomic profiling using proton nuclear magnetic resonance (1H NMR) spectroscopy and multivariate data analysis of human serum samples was used to characterize metabolic profiles in renal cell carcinoma (RCC). We found distinct, easily detectable differences between (a) RCC patients and healthy humans, (b) RCC patients with metastases and without metastases, and (c) RCC patients before and after nephrectomy. Compared to healthy human serum, RCC serum had higher levels of lipid (mainly very low-density lipoproteins), isoleucine, leucine, lactate, alanine, N-acetylglycoproteins, pyruvate, glycerol, and unsaturated lipid, together with lower levels of acetoacetate, glutamine, phosphatidylcholine/choline, trimethylamine-N-oxide, and glucose. This pattern was somewhat reversed after nephrectomy. Altered metabolite concentrations are most likely the result of the cells switching to glycolysis to maintain energy homeostasis following the loss of ATP caused by impaired TCA cycle in RCC. Serum NMR spectra combined with principal component analysis techniques offer an efficient, convenient way of depicting tumour biochemistry and stratifying tumours under different pathophysiological conditions. It may be able to assist early diagnosis and postoperative surveillance of human malignant diseases using single blood samples

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

    Muhammad Tayyab Akhtar

    2016-08-01

    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.

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

    Mohamed M. Youssef

    2011-05-01

    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.

  6. A Comparative Study of the Organic Acid Content of the Hemolymph of Schistosoma mansoni-Resistant and Susceptible Strains of Biomphalaria glabrata

    José Clecildo Barreto Bezerra

    1997-05-01

    Full Text Available The freshwater snail Biomphalaria glabrata is an intermediate host of the trematode Schistosoma mansoni. However, some strains of B. glabrata are resistant to successful infection by S. mansoni larvae. The present work examines the profile of organic acids present in S. mansoni-resistant and -susceptible strains of B. glabrata, in order to determine whether the type of organic acid present is related to susceptibility. The organic acids were extracted from the hemolymph of two susceptible B. glabrata strains (PR, Puerto Rico and Ba, Jacobina-Bahia from Brazil, and from the resistant strains 13-16-R1 and 10R2, using solid phase extraction procedures followed by high performance liquid chromatography. The organic acids obtained were analyzed and identified by comparison with known standards. Pyruvate, lactate, succinate, malate, fumarate, acetate, propionate, ß-hydroxybutyrate and acetoacetate were detected in all hemolymph samples. Under standard conditions, the concentration of each of these substances varied among the strains tested and appeared to be specific for each strain. An interesting variation was the low concentration of pyruvate in the hemolymph of PR-snails. Only the concentration of fumarate was consistently different (p£ 0.05 between resistant and susceptible strains

  7. Dark fermentation: isolation and characterization of hydrogen-producing strains from sludges.

    Rajhi, Haifa; Conthe, Mónica; Puyol, Daniel; Díaz, Emiliano; Sanz, José Luis

    2013-03-01

    To improve bacterial hydrogen production, ten hydrogen-producing strains belonging to Clostridium spp. were isolated from various sludges under low vacuum. Hydrogenogenesis by dark fermentation in batch cultures of these strains was optimal at about 35 degrees C and an initial pH of 6.5, which for all strains gradually dropped to ca. pH 4 during the fermentation. Clostridium roseum H5 and C. diolis RT2 had the highest hydrogen yields per total substrate (120 ml H2/g initial COD). Substrate consumption alone by C. beijerinckii UAM and C. diolis RT2 reached 573 and 475 ml H2/g consumed COD, respectively. Butyric acid fermentation was predominant, with butyrate and acetate as the major by-products and propionate, ethanol, and lactate as secondary metabolites. The acetate:butyrate ratios and fermentation pathways varied depending on the strains and environmental conditions. Hydrogenogenesis was studied in greater detail in C. saccharobutylicum H1. In butyric acid fermentation by this representative strain, acetoacetate was detected as an intermediate metabolite. Hydrogenogenesis was also analyzed in an enrichment culture, which behaved similarly to the axenic cultures. PMID:24151782

  8. Benzothiazolylazo derivatives of some β-dicarbonyl compounds and their Cu(II, Ni(II and Zn(II complexes

    KRISHNANNAIR KRISHNANKUTTY

    2010-05-01

    Full Text Available The coupling of diazotized 2-aminobenzothiazole with 1,3-dicar-bonyl compounds (benzoylacetone, methyl acetoacetate and acetoacetanilide yielded a new series of tridentate ligand systems (HL. Analytical, IR, 1H--NMR, 13C-NMR and mass spectral data indicated that the compounds exist in the intramolecularly hydrogen bonded azo-enol tautomeric form in which one of the carbonyl groups of the dicarbonyl moiety had enolised and hydrogen bonded to one of the azo nitrogen atoms. The compounds formed stable complexes with Ni(II, Cu(II and Zn(II ions. The Cu(II complexes conform to [CuL(OAc] stoichiometry while the Ni(II and Zn(II complexes are in agreement with [ML2] stoichiometry. Analytical, IR, 1H-NMR, 13C-NMR and mass spectral data of the complexes are consistent with the replacement of the chelated enol proton of the ligand with a metal ion, thus leading to a stable six-membered chelate ring involving a cyclic nitrogen, one of the azo nitrogens and the enolate oxygen. The Zn(II chelates are diamagnetic while Cu(II and Ni(II complexes showed a normal paramagnetic moment.

  9. Acetone formation in the Vibrio family: a new pathway for bacterial leucine catabolism.

    Nemecek-Marshall, M; Wojciechowski, C; Wagner, W P; Fall, R

    1999-12-01

    There is current interest in biological sources of acetone, a volatile organic compound that impacts atmospheric chemistry. Here, we determined that leucine-dependent acetone formation is widespread in the Vibrionaceae. Sixteen Vibrio isolates, two Listonella species, and two Photobacterium angustum isolates produced acetone in the presence of L-leucine. Shewanella isolates produced much less acetone. Growth of Vibrio splendidus and P. angustum in a fermentor with controlled aeration revealed that acetone was produced after a lag in late logarithmic or stationary phase of growth, depending on the medium, and was not derived from acetoacetate by nonenzymatic decarboxylation in the medium. L-Leucine, but not D-leucine, was converted to acetone with a stoichiometry of approximately 0.61 mol of acetone per mol of L-leucine. Testing various potential leucine catabolites as precursors of acetone showed that only alpha-ketoisocaproate was efficiently converted by whole cells to acetone. Acetone production was blocked by a nitrogen atmosphere but not by electron transport inhibitors, suggesting that an oxygen-dependent reaction is required for leucine catabolism. Metabolic labeling with deuterated (isopropyl-d(7))-L-leucine revealed that the isopropyl carbons give rise to acetone with full retention of deuterium in each methyl group. These results suggest the operation of a new catabolic pathway for leucine in vibrios that is distinct from the 3-hydroxy-3-methylglutaryl-coenzyme A pathway seen in pseudomonads. PMID:10601206

  10. Isotopomer Spectral Analysis: Utilizing Nonlinear Models in Isotopic Flux Studies.

    Kelleher, Joanne K; Nickol, Gary B

    2015-01-01

    We present the principles underlying the isotopomer spectral analysis (ISA) method for evaluating biosynthesis using stable isotopes. ISA addresses a classic conundrum encountered in the use of radioisotopes to estimate biosynthesis rates whereby the information available is insufficient to estimate biosynthesis. ISA overcomes this difficulty capitalizing on the additional information available from the mass isotopomer labeling profile of a polymer. ISA utilizes nonlinear regression to estimate the two unknown parameters of the model. A key parameter estimated by ISA represents the fractional contribution of the tracer to the precursor pool for the biosynthesis, D. By estimating D in cells synthesizing lipids, ISA quantifies the relative importance of two distinct pathways for flux of glutamine to lipid, reductive carboxylation, and glutaminolysis. ISA can also evaluate the competition between different metabolites, such as glucose and acetoacetate, as precursors for lipogenesis and thereby reveal regulatory properties of the biosynthesis pathway. The model is flexible and may be expanded to quantify sterol biosynthesis allowing tracer to enter the pathway at three different positions, acetyl CoA, acetoacetyl CoA, and mevalonate. The nonlinear properties of ISA provide a method of testing for the presence of gradients of precursor enrichment illustrated by in vivo sterol synthesis. A second ISA parameter provides the fraction of the polymer that is newly synthesized over the time course of the experiment. In summary, ISA is a flexible framework for developing models of polymerization biosynthesis providing insight into pools and pathway that are not easily quantified by other techniques. PMID:26358909

  11. Keto-acids in Carbonaceous Meteorites

    Cooper, G.; Chang, P. M.; Dugas, A.; Byrd, A.; Chang, P. M.; Washington, N.

    2005-01-01

    The Murchison and Murray meteorites are the best-characterized carbonaceous meteorites with respect to organic chemistry and are generally used as references for organic compounds in extraterrestrial material. Among the classes of organic compounds found in these meteorites are amino acids, carboxylic acids, hydroxy acids, purines, and pyrimidines. Such compounds, important in contemporary biochemistry, are thought to have been delivered to the early Earth in asteroids and comets and may have played a role in early life and/or the origin of life. Absent among (today's) critically important biological compounds reported in carbonaceous meteorites are keto acids, i.e., pyruvic acid, acetoacetic acid, and higher homologs. These compounds are key intermediates in such critical processes as glycolysis and the citric acid cycle. In this study several individual meteoritic keto acids were identified by gas chromatography-mass spectrometry (GC-MS) (see figure below). All compounds were identified as their trimethylsilyl (TMS), isopropyl ester (ISP), and tert-butyldimethylsilyl (tBDMS) derivatives. In general, the compounds follow the abiotic synthesis pattern of other known meteorite classes of organic compounds [1,2]: a general decrease in abundance with increasing carbon number within a class of compounds and many, if not all, possible isomers present at a given carbon number. The majority of the shown compounds was positively identified by comparison of their mass spectra to commercially available standards or synthesized standards.

  12. Suggested involvement of ketone bodies in the pathogenesis of the metabolic syndrome.

    Alexandre, Adolfo

    2013-05-01

    Untreated brain mitochondria are strong producers of H2O2. High peroxide production (in the presence of glutamate and pyruvate) is strictly succinate-dependent. Importantly, it is inhibited by the ketone body acetoacetate (AcAc) starting at 10 μM (maximal effect at 0.5mM). Butyrate derives from the fermentation of prebiotics, is present physiologically in the colon and is a strong producer of AcAc: indeed butyrate induces in the colon the transcription of mitochondrial 3-hydroxy-3-methyl glutarylCoA (HMGCoA) synthase, a key enzyme in ketone body synthesis. Obesity and insulin resistance were shown to be dependent on increased permeability of the colon epithelium to bacterial lipopolysaccharide (LPS); the process is evident particularly upon ingestion of lipids (a peroxidative event, inhibited by vitamin E) and is likely sensitive to AcAc. The oxidation of butyrate and the production of AcAc in the colon appear to be inhibited by high luminal sulphides and high NH3, a situation that presumably facilitates LPS permeation (on the contrary beta-hydroxy-butyrate oxidation is not inhibited). It is proposed that these damaging events may be opposed by the delivery of ketone bodies directly to the colon. PMID:23466063

  13. Stereoselective Reduction of Prochiral Ketones by Plant and Microbial Biocatalysts

    Javidnia, K.; Faghih-Mirzaei, E.; Miri, R.; Attarroshan, M.; Zomorodian, K.

    2016-01-01

    Chiral alcohols are the key chiral building blocks to many enantiomerically pure pharmaceuticals. The biocatalytic approach in asymmetric reduction of corresponding prochiral ketones to the preparation of these optically pure substances is one of the most promising routes. The stereoselective reduction of different kinds of prochiral ketones catalyzed by various plants and microorganisms was studied in this work. Benzyl acetoacetate, methyl 3-oxopentanoate, ethyl 3-oxopentanoate, and ethyl butyryl acetate were chosen as the model substrates for β-ketoesters. Benzoyl acetonitrile, 3-chloro propiophenone, and 1-acetyl naphthalene were chosen as aromatic aliphatic ketones. Finally, 2-methyl benzophenone and 4-chloro benzophenone were selected as diaryl ketones. Plant catalysis was conducted by Daucus carota, Brassica rapa, Brassica oleracea, Pastinaca sativa, and Raphnus sativus. For microbial catalysis, Aspergillus foetidus, Penicillum citrinum, Saccharomyces carlbergensis, Pichia fermentans, and Rhodotrula glutinis were chosen. Chiral alcohols were obtained in high yields and with optical purity. A superiority in the microorganisms' performance in the bioreduction of prochiral ketones was detected. Among microorganisms, Rhodotrula glutinis showed remarkable results with nearly all substrates and is proposed for future studies.

  14. Michael addition reaction catalyzed by solid base%用固体碱催化Michael加成反应的研究

    徐景士; 王红明; 陈慧宗

    2003-01-01

    Different Michael addition reactions catalyzed by solid base K2O/γ-Al2O3 and KF/γ-Al2O3, MgO/γ-Al2O3 prepared by microwave irradiation method were reported in this paper. For the K2O/γ-Al2O3, not only good yield was attained but also made the reaction of Acetonitrile that usually is regarded as inactive carbonion and α, β-unsatumted compound to carry out. The yield of Michael reaction of Ethyl acetoacetate and Crotonaldehyde catalyzed by KF/γ-Al2O3, MgO/γ-Al2O3, MgO/NaY all could reach to 80% and the latter could reach to 90%. At the same time, the catalysts of different content(the ratio of load) of MgO were also applied in this reaction, and it was found that the best content of MgO was 20% ~ 25%。

  15. "Synthesis and smooth muscle Calcium channel antagonist effect of Alkyl, Aminoalkyl 1,4-Dihydro-2,6-Dimethyl-4-Nitroimidazole-3,5 Pyridine Dicarboxylates "

    Miri R

    2001-08-01

    Full Text Available The discovery that 1,4-dihydropyridine (DHP class of calcium channel antagonist inhibits the Ca+² influx represented a major therapeutic advance in the treatment of cardiovascular diseases such as hypertension, angina pectoris and other spastic smooth muscle disorders. A novel class of calcium channel antagonist of flunarizine containing arylpiperazinyl moiety has recently been reported. It was therefore of interest to determine the effect that selected C-3 substituents contained amino alkyl and arylpiperazine, in conjunction with a C-4 1-methyl-5-nitro-2-imidazolyl substituents on calcium channel antagonist activity. The unsymmetrical analogues were prepared by a procedure reported by Meyer in which 1-methyl-5-nitro-imidazol-2-carboxaldehyde was reacted with acetoacetic esters and alkyl 3-aminocrotonate. In vitro calcium channel antagonist activities were determined by the use of high K+ contraction of guinea pig ileal longitudinal smooth muscle. All compounds exhibited comparable calcium channel antagonist activity (IC50=10^-9 to 10^-11 M against reference drug nifedipine (IC50=2.75±0.36 x 10^-10 M.

  16. 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: safaei@kashanu.ac.ir [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)

    2015-08-15

    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.

  17. 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

    2016-01-01

    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. PMID:27517894

  18. Gender-specific metabolomic profiling of obesity in leptin-deficient ob/ob mice by 1H NMR spectroscopy.

    Eun-Young Won

    Full Text Available Despite the numerous metabolic studies on obesity, gender bias in obesity has rarely been investigated. Here, we report the metabolomic analysis of obesity by using leptin-deficient ob/ob mice based on the gender. Metabolomic analyses of urine and serum from ob/ob mice compared with those from C57BL/6J lean mice, based on the (1H NMR spectroscopy in combination with multivariate statistical analysis, revealed clear metabolic differences between obese and lean mice. We also identified 48 urine and 22 serum metabolites that were statistically significantly altered in obese mice compared to lean controls. These metabolites are involved in amino acid metabolism (leucine, alanine, ariginine, lysine, and methionine, tricarbocylic acid cycle and glucose metabolism (pyruvate, citrate, glycolate, acetoacetate, and acetone, lipid metabolism (cholesterol and carnitine, creatine metabolism (creatine and creatinine, and gut-microbiome-derived metabolism (choline, TMAO, hippurate, p-cresol, isobutyrate, 2-hydroxyisobutyrate, methylamine, and trigonelline. Notably, our metabolomic studies showed distinct gender variations. The obese male mice metabolism was specifically associated with insulin signaling, whereas the obese female mice metabolism was associated with lipid metabolism. Taken together, our study identifies the biomarker signature for obesity in ob/ob mice and provides biochemical insights into the metabolic alteration in obesity based on gender.

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

    Zhang, Yifan

    2012-06-15

    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.

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

    E.R. El-Sawy

    2014-12-01

    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.

  1. Synthesis of Clevidipine Butyrate%氯维地平的合成

    张婧; 纪宪勇; 孙翔; 王杰

    2011-01-01

    Clevidipine butyrate, an antihypertensive agent, was synthesized from 2-cyanoethyl acetoacetate (2), 2,3-dichlorobenzaldehyde and methyl 3-aminocrotonate by Hantzsch cyclocondensation and followed by selective hydrolysis with sodium sulfide at room temperature to give 4-(2,3-dichlorophenyl)-l,4-dihydro-2,6-dimethyl-3,5-pyridinedicarboxylic acid monomethyl ester, which was then subjected to reaction with chloromethyl butyrate with an overall yield of about 59% (based on 2).%3-羟基丙腈和双乙烯酮在三乙胺作用下制得乙酰乙酸(2ˉ氰基乙基)酯(2),再与2,3-二氯苯甲醛和3ˉ氨基巴豆酸甲酯经Hantzsch缩合闭环,接着用硫化钠在常温下选择性水解得4-(2,3-二氯苯基)-1-4-二氢-2,6-二甲基-3,5-吡啶二羧酸单甲酯,最后与正丁酸氯甲酯反应即得抗高血压药氯维地平,总收率约59%(以2计).

  2. Serum metabolic signature of minimal hepatic encephalopathy by (1)H-nuclear magnetic resonance.

    Jiménez, Beatriz; Montoliu, Carmina; MacIntyre, David A; Serra, Miguel A; Wassel, Abdallah; Jover, María; Romero-Gomez, Manuel; Rodrigo, Jose M; Pineda-Lucena, Antonio; Felipo, Vicente

    2010-10-01

    Minimal hepatic encephalopathy (MHE) reduces quality of life of cirrhotic patients, predicts overt hepatic encephalopathy, and is associated with poor prognosis. We hypothesized that MHE arises once metabolic alterations derived from the liver reach a particular threshold. Our aim was to assess whether metabolic profiling of serum samples by high-field (1)H-nuclear magnetic resonance spectroscopy ((1)H NMR) and subsequent multivariate analyses would be useful to characterize metabolic perturbations associated with MHE and to identify potential metabolic biomarkers. Metabolic serum profiles from controls (n = 69) and cirrhotic patients without MHE (n = 62) and with MHE (n = 39) were acquired using high field NMR. Supervised modeling of the data provided perfect discrimination between healthy controls and cirrhotic patients and allowed the generation of a predictive model displaying strong discrimination between patients with and without MHE (R(2)Y = 0.68, Q(2)Y = 0.63). MHE patients displayed increased serum concentrations of glucose, lactate, methionine, TMAO, and glycerol, as well as decreased levels of choline, branch amino acids, alanine, glycine, acetoacetate, NAC, and lipid moieties. Serum metabonomics by (1)H NMR offers a useful approach for characterizing underlying metabolic differences between patients with and without MHE. This procedure shows great potential as a diagnostic tool of MHE as it objectively reflects measurable biochemical differences between the patient groups and may facilitate monitoring of both disease progression and effects of therapeutic treatments. PMID:20690770

  3. Imact of feeding and post prandial time on plasma ketone bodies in sows during transition and lactation

    Theil, Peter Kappel; Olesen, A K; Flummer, Christine;

    2013-01-01

    high levels (3% or 8%) of dietary fats with different proportions of medium- and long-chain fatty acids. Blood was obtained by jugular venipuncture on d 3 and 7 prepartum, and d 1, 10, 17, and 28 postpartum, and concentrations of plasma beta-hydroxy butyric acid (BHBA), acetoacetate + acetone (Ac......Ac+Ac), glucose, NEFA, lactate, acetate, and butyrate were determined. For 4 out of 5 treatments, plasma BHBA decreased slightly, whereas plasma AcAc+Ac remained stable. However, plasma BHBA (P < 0.01) and AcAc+Ac (P < 0.001) doubled after d 10 of lactation in sows fed 4% octanoic acid and 4% fish oil diet (4......+4% FO; P < 0.001), compared with earlier in lactation (P < 0.001). Plasma AcAc+Ac was positively related to BHBA (P < 0.01), glucose (P < 0.05), and butyrate (P < 0.001), and negatively related to the acetate:butyrate ratio (P < 0.001). In addition, plasma BHBA was positively related to lactate (P < 0...

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

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

    2013-09-01

    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.

  5. The stability of free radicals in densely crosslinked acrylate polymer networks

    Four types of crosslinked polymeric systems based on pentaerythritetatraacrylate (PETA) and propylene glycol - 424 -diacetonate (PGDSAA) were studied from the standpoints of free radical decay and the network density. The networks samples were prepared by two step synthesis. At first the Michael dark reaction results for multifunctional acrylates and acetoacetates in polymer network. The crosslinking continued in the second step of network formation by a photo-polymerization of the residual vinyl monomer, and pendant double bonds build up in the primary network in the first step of the synthesis. The free radical accumulation to the steady state during the irradiation, and their decay in the dark periods were studied directly in ESR spectrometer at 25 grad C. The kinetic of the radicals decay after switch of the light was used to calculate the rate constants for bimolecular termination reaction kt. These values show on the existence of long lived free radicals in the investigated polymeric networks. The stability of radicals is approximately of two decimal orders higher in networks irradiated in nitrogen compared to irradiation in air. A loss of the hyperfine structure of the spectra during repeated irradiation cycles is explained by increasing spin-spin dipolar interactions when radicals are trapped in more rigid regions of the polymer matrix. (authors)

  6. Changes of arterial blood ketone body ratio following hypoperfusion in old and adult rats

    Ling YE; Shiwen WANG; Songtao YU; Wei CHEN

    2004-01-01

    Objective To evaluate the sensitivity of arterial ketone body ratio as an indicator for multiple organ failure.Materials and methods The experimental model of multiple organ failure was made in adult and old rats by hypoperfusion-induced hemorrhagic shock. After blood sampling, the arterial acetoacetate, β-hydroxybutyrate, total ketone body, ALT, AST, BUN, creatinine at 2, 4, 8 hr in hypoperfusion were examined to compare the differences of ketone body ratio and organ failure between adult and old rats. Hepatic and mitochondrial metabolism were assessed by comparing ketone body ratios (AcAc/β-OHB) and free NAD+/NADH ratios. Results Ketone body ratio in old rats at 2, 4, 8 hr after the induction of hemorrhagic shock decreased from 0.68 to 0.31, 0.27 and 0.22, respectively. In adult rats, it decreased from 1.12 to 0.17, 0.12 and 0.09, respectively. Changes of ketone body ratio in the adult group were larger than in the elderly group ( P < 0.001). The development of multiple organ failure is associated with the time of hemorrhagic shock development. Conclusions There was a different ketone body ratio between multiple organ failure in the elderly (MOFE) and multiple organ failure (MOF) in general adults. Ketone body ratio is a better indicator than ALT and AST in reflecting hepatic function in the early status of MOF. (J Geriatr Cardiol 2004;1(2) :125-128. )

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

    Anita Krishankant Taksande

    2015-12-01

    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: http://dx.doi.org/10.17807/orbital.v7i4.633 

  8. Can ketones compensate for deteriorating brain glucose uptake during aging? Implications for the risk and treatment of Alzheimer's disease.

    Cunnane, Stephen C; Courchesne-Loyer, Alexandre; St-Pierre, Valérie; Vandenberghe, Camille; Pierotti, Tyler; Fortier, Mélanie; Croteau, Etienne; Castellano, Christian-Alexandre

    2016-03-01

    Brain glucose uptake is impaired in Alzheimer's disease (AD). A key question is whether cognitive decline can be delayed if this brain energy defect is at least partly corrected or bypassed early in the disease. The principal ketones (also called ketone bodies), β-hydroxybutyrate and acetoacetate, are the brain's main physiological alternative fuel to glucose. Three studies in mild-to-moderate AD have shown that, unlike with glucose, brain ketone uptake is not different from that in healthy age-matched controls. Published clinical trials demonstrate that increasing ketone availability to the brain via moderate nutritional ketosis has a modest beneficial effect on cognitive outcomes in mild-to-moderate AD and in mild cognitive impairment. Nutritional ketosis can be safely achieved by a high-fat ketogenic diet, by supplements providing 20-70 g/day of medium-chain triglycerides containing the eight- and ten-carbon fatty acids octanoate and decanoate, or by ketone esters. Given the acute dependence of the brain on its energy supply, it seems reasonable that the development of therapeutic strategies aimed at AD mandates consideration of how the underlying problem of deteriorating brain fuel supply can be corrected or delayed. PMID:26766547

  9. 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

    1996-04-01

    3-hydroxy-3-methylglutaryl CoA lyase (HL, EC 4.1.3.4) 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.

  10. Immunoaffinity purification and characterization of mitochondrial membrane-bound D-3-hydroxybutyrate dehydrogenase from Jaculus orientalis

    Cherkaoui-Malki Mustapha

    2008-09-01

    Full Text Available Abstract Background The interconversion of two important energy metabolites, 3-hydroxybutyrate and acetoacetate (the major ketone bodies, is catalyzed by D-3-hydroxybutyrate dehydrogenase (BDH1: EC 1.1.1.30, a NAD+-dependent enzyme. The eukaryotic enzyme is bound to the mitochondrial inner membrane and harbors a unique lecithin-dependent activity. Here, we report an advanced purification method of the mammalian BDH applied to the liver enzyme from jerboa (Jaculus orientalis, a hibernating rodent adapted to extreme diet and environmental conditions. Results Purifying BDH from jerboa liver overcomes its low specific activity in mitochondria for further biochemical characterization of the enzyme. This new procedure is based on the use of polyclonal antibodies raised against BDH from bacterial Pseudomonas aeruginosa. This study improves the procedure for purification of both soluble microbial and mammalian membrane-bound BDH. Even though the Jaculus orientalis genome has not yet been sequenced, for the first time a D-3-hydroxybutyrate dehydrogenase cDNA from jerboa was cloned and sequenced. Conclusion This study applies immunoaffinity chromatography to purify BDH, the membrane-bound and lipid-dependent enzyme, as a 31 kDa single polypeptide chain. In addition, bacterial BDH isolation was achieved in a two-step purification procedure, improving the knowledge of an enzyme involved in the lipid metabolism of a unique hibernating mammal. Sequence alignment revealed conserved putative amino acids for possible NAD+ interaction.

  11. False-positive breath-alcohol test after a ketogenic diet.

    Jones, A W; Rössner, S

    2007-03-01

    A 59-year-old man undergoing weight loss with very low calorie diets (VLCD) attempted to drive a car, which was fitted with an alcohol ignition interlock device, but the vehicle failed to start. Because the man was a teetotaller, he was surprised and upset by this result. VLCD treatment leads to ketonemia with high concentrations of acetone, acetoacetate and beta-hydroxybutyrate in the blood. The interlock device determines alcohol (ethanol) in breath by electrochemical oxidation, but acetone does not undergo oxidation with this detector. However, under certain circumstances acetone is reduced in the body to isopropanol by hepatic alcohol dehydrogenase (ADH). The ignition interlock device responds to other alcohols (e.g. methanol, n-propanol and isopropanol), which therefore explains the false-positive result. This 'side effect' of ketogenic diets needs further discussion by authorities when people engaged in safety-sensitive work (e.g. bus drivers and airline pilots) submit to random breath-alcohol tests. PMID:16894360

  12. Comparison of palmitic acid kinetics during glucose or ketone body infusions

    Birkhahn, R.H.; Block, D.J.; Birkhahn, G.C.; Thomford, N.R.

    1986-03-05

    Ketone body interactions can be observed for extended ketosis by infusion by monoacetoacetin (the monoglyceride of acetoacetic acid). Palmitic acid kinetics were compared on the 5th day of glucose or ketone body-glucose infusions. 20 rats were fed complete diets intravenously at the rate of 50 ml/day. All diets contained vitamins, trace minerals, electrolytes, amino acids and 1 kcal/ml of non-protein energy. Rats were divided by energy source: Group A (n = 10) received energy from glucose and Group B (n = 10) from 72% monoacetoacetin plus 28% glucose. Diets were given at 1/2 and 3/4 rats on days 1 and 2, respectively and at full rate for days 3-5. Urinary nitrogen losses, body weight and dietary intake were measured daily. Palmitate kinetics was measured on day 5 using a continuous infusion of (1-/sup 14/C) palmitate and measuring C-14 in breath and plasma and plasma palmitate by GC. The two groups had similar body weight changes and urinary nitrogen losses over the 3 days of full intake Group A had lower plasma palmitate (88 +/- 7 vs 105 +/- 6 micromol/l) but similar turnover (17.1 +/- 2.4 vs 15.0 +/- 1.9 mmol/hr) and oxidation 2.3 +/- 0.3 vs 2.2 +/- 0.05 mmol/hr) compared to Group B. These data show that feeding monoacetoacetin intravenously does not stimulate fatty acid metabolism in the well nourished rat.

  13. Serum and urine trace metals in ketosis

    Burch, R.E.; Hiner, C.; Sullivan, J.F.

    1974-01-01

    Three groups of male rats were housed in metabolic cages. The control group was fed ad libitum, a second group was fasted and diabetes was induced in the third group by the subcutaneous injection of alloxan. After 48 hours all animals were bled and livers, which were quick-frozen in situ, were obtained. Fasted and diabetic animals were ketotic as manifested by lowering urinary pH and elevated urinary and blood acetoacetate and ..beta..-hydroxybutyrate. Blood and hepatic content of acetate was elevated in diabetic versus control rats. Blood and hepatic content of acetate was not increased in fasted animals. In fasted rats total urinary Zn and Cu were decreased and Mg and Ca did not differ significantly from control values. In diabetic rats total urinary excretion of Mg was increased whereas urinary Zn, Cu and Ca levels did not differ significantly from control values. In fasted animals only serum Zn was diminished. In diabetic rats serum Cu and Ca were decreased and serum Mg was increased while serum Zn was unchanged. Thus, the ketosis of fasting differs from the ketosis of diabetes in several of its metabolic manifestations. 11 references, 9 figures.

  14. Breath Ketone Testing: A New Biomarker for Diagnosis and Therapeutic Monitoring of Diabetic Ketosis

    Yue Qiao

    2014-01-01

    Full Text Available Background. Acetone, β-hydroxybutyric acid, and acetoacetic acid are three types of ketone body that may be found in the breath, blood, and urine. Detecting altered concentrations of ketones in the breath, blood, and urine is crucial for the diagnosis and treatment of diabetic ketosis. The aim of this study was to evaluate the advantages of different detection methods for ketones, and to establish whether detection of the concentration of ketones in the breath is an effective and practical technique. Methods. We measured the concentrations of acetone in the breath using gas chromatography-mass spectrometry and β-hydroxybutyrate in fingertip blood collected from 99 patients with diabetes assigned to groups 1 (−, 2 (±, 3 (+, 4 (++, or 5 (+++ according to urinary ketone concentrations. Results. There were strong relationships between fasting blood glucose, age, and diabetic ketosis. Exhaled acetone concentration significantly correlated with concentrations of fasting blood glucose, ketones in the blood and urine, LDL-C, creatinine, and blood urea nitrogen. Conclusions. Breath testing for ketones has a high sensitivity and specificity and appears to be a noninvasive, convenient, and repeatable method for the diagnosis and therapeutic monitoring of diabetic ketosis.

  15. Catalytic synthesis of 5-ethoxycarbonyl-4-(4-hydroxyphenyl)-6-methyl-3,4-dihydropyrimidine-2(1H)-ketone with H6P2W18O62/SiO2%H6P2W18O62/SiO2催化合成5-乙氧羰基-4-(4-羟基苯基)-6-甲基-3,4-二氢嘧啶-2(1H)-酮

    刘晓霞; 肖欢; 杨水金

    2015-01-01

    5-Ethoxycarbonyl-4-(4-hydroxyphenyl)-6-methyl-3,4-dihydropyrimidine-2(1H)-ketone was synthesized by using H6P2W18O62/SiO2 as the catalyst and p-hydroxy benzaldehyde,ethyl acetoacetate and urea as raw materials. Catalytic activity of this reaction using H6P2W18O62/SiO2 as the catalyst was explored. The effects of reaction on the yield of the product were investigated and the optimum conditions were determined as follows:fixing p-hydroxy benzaldehyde consumption is 0.04 mol,n(p-hydroxy benzaldehyde):n(ethyl acetoacetate):n(urea)=1∶1.5∶1.5,the mass ratio of the catalyst to total reactant is 1.5%,the reaction temperature is 80℃, and the reaction time is 1.0 h. The yield of the product was up to 94.4%.%采用H6P2W18O62/SiO2为催化剂,以对羟基苯甲醛、乙酰乙酸乙酯、尿素为原料合成5-乙氧羰基-4-(4-羟基苯基)-6-甲基-3,4-二氢嘧啶-2(1H)-酮.探讨H6P2W18O62/SiO2对本反应的催化活性,较系统地研究了反应物物质的量比、催化剂用量、反应时间、反应温度四因素对产物收率的影响.实验表明:H6P2W18O62/SiO2是合成5-乙氧羰基-4-(4-羟基苯基)-6-甲基-3,4-二氢嘧啶-2(1H)-酮的良好催化剂;在n (对羟基苯甲醛)∶n (乙酰乙酸乙酯)∶n (尿素)=1∶1.5∶1.5,催化剂的用量占反应物料总质量的1.5%,反应温度为80℃,反应时间为60 min的最佳条件下,其收率可达94.4%.

  16. Assessment of Early Postpartum Reproductive Performance in Two High Producing Estonian Dairy Herds

    Kindahl H

    2003-09-01

    Full Text Available Early postpartum (6 weeks ovarian activity, hormonal profiles, uterine involution, uterine infections, serum electrolytes, glucose, milk acetoacetate and blood urea nitrogen (BUN levels were studied in 2 Estonian high producing dairy herd with annual milk production of 7688 (Farm A and 9425 (Farm B. From each farm 10 cows, with normal calving performance were used. Blood samples for the hormonal (PGF2α-metabolite, progesterone analyses were withdrawn. On day 25 PP blood serum samples were taken for the evaluation of metabolic/electrolyte status. On the same day estimation of milk acetoacetate values was done. The ultrasound (US was started on day 7 PP and was performed every 3rd day until the end of experiment. Uterine content, follicular activity and sizes of the largest follicle and corpus luteum were monitored and measured. Vaginal discharge and uterine tone were recorded during the rectal palpation. Each animal in the study was sampled for bacteriological examination using endometrial biopsies once a week. Two types of PGF2α-metabolite patterns were detected: elevated levels during 14 days PP, then decline to the basal level and then a second small elevation at the time of final elimination of the bacteria from the uterus; or elevated levels during first 7 days PP, then decline to the basal level and a second small elevation before the final elimination of bacteria. Endometritis was diagnosed in 5 cows in farm A and in 3 cows in farm B respectively. In farm A, 5 cows out of 10 ovulated during experimental period and in 1 cow cystic ovaries were found. In farm B, 3 cows out of 10 ovulated. In 3 cows cystic ovaries were found. Altogether 40% of cows had their first ovulation during the experimental period. Three cows in farm A and 5 cows in farm B were totally bacteria negative during the experimental period. The most frequent bacteria found were A. pyogenes, Streptococcus spp., E. coli., F. necrophorum and Bacteroides spp. The highest

  17. Applications of lasers in metallization of thermoplastic and thermosetting polymers

    P. Rytlewski

    2013-04-01

    Full Text Available Purpose: This work focuses on the studies of chemical and physical changes induced by ArF-laser irradiation leading to formation of surfaces catalytically highly active and fully prepared for the direct electroless metallization for the case of thermoplastic and thermosetting polymer composites. The only pretreatment method for surface to be activated was laser irradiation. There are compared two polymer composites: thermoplastic and thermosetting with the same qualitative and quantitative contents of the selected copper compounds. Additionally, there is presented wide context of laser applications in electroless metallization of polymeric materials.Design/methodology/approach: The composites contained the same amount of copper(II oxide (CuO and copper(II acetoacetate Cu(acac2, while varied with the type of polymer matrix. There were chosen polyamide 6 as thermoplastic and polyurethane resin as thermosetting polymer matrixes. The composites were irradiated with various numbers of ArF excimer laser pulses (λ = 193 nm at constant fluence of 100 mJ/cm2. The metallization procedure of the laser-irradiated samples was performed by use of a commercial metallization bath and formaldehyde as a reducing agent. The samples were examined using FTIR, contact angle measurement and SEM techniques.Findings: It was found that laser irradiation induce catalytic properties in the studied composites. However, better catalytic properties were achieved for the thermoplastic than thermosetting polymer composites.Research limitations/implications: In order to better understand the differences in laser interactions between thermoplastic and thermosetting composites more examples of various polymer matrixes should be investigated.Practical implications: Suitable condition for laser irradiation of the composites associated with the best catalytic properties were proposes. Better catalytic properties were achieved for thermoplastic than thermosetting composite

  18. 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

    尹显洪

    2003-01-01

      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.

  19. Isopropanol production with engineered Cupriavidus necator as bioproduction platform

    Grousseau, E; Lu, JN; Gorret, N; Guillouet, SE; Sinskey, AJ

    2014-03-07

    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 beta-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.

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

    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.

  1. Systemic and characteristic metabolites in the serum of streptozotocin-induced diabetic rats at different stages as revealed by a (1)H-NMR based metabonomic approach.

    Diao, Chengfeng; Zhao, Liangcai; Guan, Mimi; Zheng, Yongquan; Chen, Minjiang; Yang, Yunjun; Lin, Li; Chen, Weijian; Gao, Hongchang

    2014-03-01

    Diabetes mellitus is a typical heterogeneous metabolic disorder characterized by abnormal metabolism of carbohydrates, lipids, and proteins. Investigating the changes in metabolic pathways during the evolution of diabetes mellitus may contribute to the understanding of its metabolic features and pathogenesis. In this study, serum samples were collected from diabetic rats and age-matched controls at different time points: 1 and 9 weeks after streptozotocin (STZ) treatment. (1)H nuclear magnetic resonance ((1)H NMR)-based metabonomics with quantitative analysis was performed to study the metabolic changes. The serum samples were also subjected to clinical chemistry analysis to verify the metabolic changes observed by metabonomics. Partial least squares discriminant analysis (PLS-DA) demonstrated that the levels of serum metabolites in diabetic rats are different from those in control rats. These findings indicate that the metabolic characteristics of the two groups are markedly different at 1 and 9 weeks. Quantitative analysis showed that the levels of some metabolites, such as pyruvate, lactate, citrate, acetone, acetoacetate, acetate, glycerol, and valine, varied in a time-dependent manner in diabetic rats. These results suggest that serum metabolites related to glycolysis, the tricarboxylic acid cycle, gluconeogenesis, fatty acid β-oxidation, branched-chain amino acid metabolism, and the tyrosine metabolic pathways are involved in the evolution of diabetes. The metabolic changes represent potential features and promote a better understanding of the mechanisms involved in the development of diabetes mellitus. This work further suggests that (1)H NMR metabonomics is a valuable approach for providing novel insights into the pathogenesis of diabetes mellitus and its complications. PMID:24448714

  2. Preparation of zirconia coatings by hydrolysis of zirconium alkoxide with hydrogen peroxide

    Zirconia has gained a great deal of attention because of its superior properties of mechanical strength, chemical resistance, and ionic conductivity. Zirconia coatings and thin films are receiving attention as tribological and thermal barrier coatings for engines, high-reflective coatings, solid electrolytes for fuel cells, oxygen sensors, etc. The sol-gel coating method has several advantages, such as low processing temperatures, homogeneity, control of micro-structure, and good productivity compared to chemical vapor deposition and physical vapor deposition. However, there are few reports concerning the preparation of zirconia coatings and thin films by the sol-gel method. Up to the present, zirconia coatings have been prepared from zirconium propoxide (not heated), zirconium tetrabutoxide modified by acetylacetone and ethyl acetoacetate, zirconium diethoxydichloride (ZrCl2(OC2H5)2), and a hydrosol prepared from a zirconium oxychloride solution. Coatings of 8.8-mol%-yttria-doped zirconia were fabricated using a transparent and spinnable sol prepared by hydrolysis of zirconium alkoxide with hydrogen peroxide and nitric acid. The sol gave a crack-free coating film consisting of fine grains. The crystalline phase was cubic after heating of 1,000 and 1,200 C and cubic and tetragonal at 1,350 C, with the coating being highly oriented in the (111) plane, especially at 1,000 C. Activation energy of the coating films was higher than that of the bulk. Transmittance through a film thickness of about 0.3 μm on each side was 75%

  3. Antioxidant, antityrosinase and antitumor activity comparison: the potential utilization of fibrous root part of Bletilla striata (Thunb. Reichb.f.

    Fusheng Jiang

    Full Text Available This study was carried out to evaluate the utilization probability of the fibrous root part (FRP of Bletilla striata, which was usually discarded and harvesting pseudobulb part (PSP. The chemical composition, total phenolic content, DPPH radical scavenging activity, Ferric-reducing antioxidant power and tyrosinase inhibition activity were compared between FRP and PSP. Antioxidant and pro-oxidant effect as well as antitumor effect of the extract of FRP and PSP were analyzed by in vitro cell system as well. Thin layer chromatography and high performance liquid chromatography analysis indicated that the chemical compositions in the two parts were similar, but the content in FRP was much higher than PSP. Meanwhile, the FRP extracts showed higher phenolic content, stronger DPPH scavenging activity, Ferric-reducing antioxidant capacity and tyrosinase inhibition activity. Sub-fraction analysis revealed that the distribution characteristic of phenolic components and other active constituents in FRP and PSP were consistent, and mainly deposited in chloroform and acetoacetate fractions. Especially, the chloroform sub-fraction (sch of FRP showed extraordinary DPPH scavenging activity and tyrosinase inhibition activity, with IC50 0.848 mg/L and 4.3 mg/L, respectively. Besides, tyrosinase inhibition activity was even stronger than the positive compound arbutin (31.8 mg/L. Moreover, In vitro cell system analysis confirmed that FRP extract exerts comparable activity with PSP, especially, the sub-fraction sch of FRP showed better antioxidant activity at low dosage and stronger per-oxidant activity at high dosage, and both sch of FRP and PSP can dose-dependent induce HepG2 cells apoptosis, which implied tumor therapeutic effect. Considering that an additional 0.3 kg FRP would be obtained when producing 1.0 kg PSP, our work demonstrated that FRP is very potential to be used together with PSP.

  4. The synthesis and characterization of 7-hydroxy-4-methylcoumarin and the investigation of the fluorescence properties of its 7-hydroxy-4-methylcoumarin-chitosan films

    Chitosan fluorescent films containing 7-hydroxy-4-methylcoumarin (7H4MC) have been successfully prepared. Used chitosan was obtained from chitin isolated from skin of tiger prawns (Penaeus monodon) through the deproteination, demineralization, and deacetylation process. The yields of chitin and chitosan are 10.66% and 23.83%, respectively. The chitosan has 55.00% degree of deacetylation based on FTIR spectroscopy. Average molecular mass of chitosan which was determined by Ostwald viscometry method is 8.55 × 106 g/mol. The 7H4MC was synthesized from resorcinol and ethyl acetoacetate using amberlyst-15 as catalyst based on Pechmann reaction with chemical yields of 90.01% and the melting point of 189–190°C. The FTIR, 1H–NMR, and 13C–NMR spectroscopies confirmed the structure which corresponds to the structure of 7H4MC. The films of chitosan containing 7H4MC were prepared by solvent evaporation method in 2% (v/v) acetic acid. The 7H4MC content in each film was 0% (blank), 0.2%, 0.4%, 0.6%, and 0.8% (w/w). The UV-Vis spectrum of 7H4MC in methanol showed λmax at 235 and 337 nm. The observed fluorescence is the fluorescence color of cyan. The excitation wavelengths are 200, 235, 275, 337, and 365 nm. The highest intensity of cyan color fluorescence of chitosan containing 7H4MC films was obtained at the concentration of 0.2% of 7-hydroxy-4-methylcoumarin at the excitation wavelength of 275 nm

  5. The accidental assignment of function in the tautomerase superfamily

    Jamison P. Huddleston

    2015-03-01

    Full Text Available Cg10062 from Corynebacterium glutamicum is a tautomerase superfamily member with the characteristic β−α−β fold and catalytic Pro-1. It is a cis-3-chloroacrylic acid dehalogenase (cis-CaaD homologue with high sequence similarity (53% that includes the six critical active site residues (Pro-1, His-28, Arg-70, Arg-73, Tyr-103, and Glu-114. However, Cg10062 is a poor cis-CaaD: it has much lower catalytic efficiency and lacks isomer specificity. Two acetylene compounds (propiolate and 2-butynoate and an allene (2,3-butadienote were investigated as potential substrates for Cg10062. Cg10062 is a hydratase/decarboxylase using propiolate and cis-3-chloro- and 3-bromoacrylates, where malonate semialdehyde is the product of hydration and acetaldehyde is the product of decarboxylation. The two activities occur consecutively using the initial substrate. In contrast, 2-butynoate and 2,3-butadienote only undergo a hydration reaction with Cg10062 to afford acetoacetate. cis-CaaD does not function as a hydratase/decarboxylase using any of these substrates, yielding only the products of hydration. Cg10062 proceeds by direct hydration or covalent catalysis (using Pro-1 depending on the substrate. Direct hydration yields the hydration products and covalent catalysis yields the hydration and decarboxylation products. Cg10062 mutants shift the reaction toward one or the other mechanism. The observation that propiolate is the best substrate suggests that Cg10062 could be a hydratase/decarboxylase in a pathway that transforms an unknown acetylene compound to acetaldehyde via propiolate. The bifunctional activity of Cg10062 might also have implications for the evolution of the dehalogenase and decarboxylase activities in the tautomerase superfamily.

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

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

    2016-08-01

    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. PMID:26928043

  7. Enhancement of L-3-hydroxybutyryl-CoA dehydrogenase activity and circulating ketone body levels by pantethine. Relevance to dopaminergic injury

    de Reggi Max

    2010-04-01

    Full Text Available Abstract Background The administration of the ketone bodies hydroxybutyrate and acetoacetate is known to exert a protective effect against metabolic disorders associated with cerebral pathologies. This suggests that the enhancement of their endogenous production might be a rational therapeutic approach. Ketone bodies are generated by fatty acid beta-oxidation, a process involving a mitochondrial oxido-reductase superfamily, with fatty acid-CoA thioesters as substrates. In this report, emphasis is on the penultimate step of the process, i.e. L-3-hydroxybutyryl-CoA dehydrogenase activity. We determined changes in enzyme activity and in circulating ketone body levels in the MPTP mouse model of Parkinson's disease. Since the active moiety of CoA is pantetheine, mice were treated with pantethine, its naturally-occurring form. Pantethine has the advantage of being known as an anti-inflammatory and hypolipidemic agent with very few side effects. Results We found that dehydrogenase activity and circulating ketone body levels were drastically reduced by the neurotoxin MPTP, whereas treatment with pantethine overcame these adverse effects. Pantethine prevented dopaminergic neuron loss and motility disorders. In vivo and in vitro experiments showed that the protection was associated with enhancement of glutathione (GSH production as well as restoration of respiratory chain complex I activity and mitochondrial ATP levels. Remarkably, pantethine treatment boosted the circulating ketone body levels in MPTP-intoxicated mice, but not in normal animals. Conclusions These finding demonstrate the feasibility of the enhancement of endogenous ketone body production and provide a promising therapeutic approach to Parkinson's disease as well as, conceivably, to other neurodegenerative disorders.

  8. 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: alancenxb@sina.com [Tianjin Children' s Hospital, Tianjin 300074 (China)

    2010-03-26

    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.

  9. Metabonomic alterations from pancreatic intraepithelial neoplasia to pancreatic ductal adenocarcinoma facilitate the identification of biomarkers in serum for early diagnosis of pancreatic cancer.

    Lin, Xianchao; Zhan, Bohan; Wen, Shi; Li, Zhishui; Huang, Heguang; Feng, Jianghua

    2016-08-16

    Pancreatic cancer is a highly malignant disease with a poor prognosis and it is essential to diagnose and treat the disease at an early stage. The aim of this study was to understand the underlying biochemical mechanisms of pancreatic intraepithelial neoplasia (PanIN) and pancreatic ductal adenocarcinoma (PDAC) and to identify potential serum biomarkers for early detection of pancreatic cancer. 7,12-Dimethylbenz(a)anthracene (DMBA)-induced PanIN and PDAC rat models were established and the serum samples were collected. The serum samples were measured using (1)H nuclear magnetic resonance (NMR) spectroscopy and analyzed by chemometric methods including principal component analysis (PCA) and (orthogonal) partial least squares discriminant analysis ((O)PLS-DA). The related biochemical pathways were derived from KEGG analysis of the significantly different metabolites. As results, some serum metabolites demonstrated alarming metabolic changes in the precursor lesion of pancreatic cancer (PanIN-2 in this study). These changes involved elevated levels of ketone compounds including 3-hydroxybutyrate, acetoacetate, and acetone, some amino acids including asparagine, glutamate, threonine, and phenylalanine, glycoproteins and lipoproteins including N-acetylglycoprotein, LDL and VLDL, and some metabolites that have been shown to contribute to mutagenicity and cancer promotion such as deoxyguanosine and cytidine. More metabolites were shown to be significantly different between PanIN and PDAC, suggesting that a more complex set of changes occurs from noninvasive precursor lesion to invasive cancer. The serum metabonomic changes of rats with PanIN and PDAC may extend our understanding of pancreatic molecular pathogenesis, and the metabolic variations from PanIN to PDAC will be helpful to understand evolution processes of the pancreatic disease. NMR-based metabonomic analysis of animal models will be beneficial for the human study and will be helpful for the early detection of

  10. 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

    2010-03-01

    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.

  11. 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: jianghua.feng@hotmail.com, E-mail: jianghua.feng@wipm.ac.cn [State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun, Jilin 130022 (China)

    2010-10-01

    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

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

    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 α-hydroxy-n-valerate, o- and p-HPA, PAG, nicotinate and hippurate accompanied by decreases in the levels of urinary α-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 (β-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 subtle

  13. Enzymatic capacities of metabolic fuel use in cuttlefish (Sepia officinalis) and responses to food deprivation: insight into the metabolic organization and starvation survival strategy of cephalopods.

    Speers-Roesch, Ben; Callaghan, Neal I; MacCormack, Tyson J; Lamarre, Simon G; Sykes, Antonio V; Driedzic, William R

    2016-08-01

    Food limitation is a common challenge for animals. Cephalopods are sensitive to starvation because of high metabolic rates and growth rates related to their "live fast, die young" life history. We investigated how enzymatic capacities of key metabolic pathways are modulated during starvation in the common cuttlefish (Sepia officinalis) to gain insight into the metabolic organization of cephalopods and their strategies for coping with food limitation. In particular, lipids have traditionally been considered unimportant fuels in cephalopods, yet, puzzlingly, many species (including cuttlefish) mobilize the lipid stores in their digestive gland during starvation. Using a comprehensive multi-tissue assay of enzymatic capacities for energy metabolism, we show that, during long-term starvation (12 days), glycolytic capacity for glucose use is decreased in cuttlefish tissues, while capacities for use of lipid-based fuels (fatty acids and ketone bodies) and amino acid fuels are retained or increased. Specifically, the capacity to use the ketone body acetoacetate as fuel is widespread across tissues and gill has a previously unrecognized capacity for fatty acid catabolism, albeit at low rates. The capacity for de novo glucose synthesis (gluconeogenesis), important for glucose homeostasis, likely is restricted to the digestive gland, contrary to previous reports of widespread gluconeogenesis among cephalopod tissues. Short-term starvation (3-5 days) had few effects on enzymatic capacities. Similar to vertebrates, lipid-based fuels, putatively mobilized from fat stores in the digestive gland, appear to be important energy sources for cephalopods, especially during starvation when glycolytic capacity is decreased perhaps to conserve available glucose. PMID:27138338

  14. The synthesis and pharmacological evaluation of (±-2,3-seco-fentanyl analogues

    LJ. DOSEN-MICOVIC

    2004-11-01

    Full Text Available An efficient, five-step synthetic approach to various acyclic 1,3-diamines has been developed and applied to the preparation of a novel class of open-chained fentanyl analogues. The acyclic derivatives 5.1–5.5 (all new compounds were synthesized with the aim of estimating the significance of the piperidine ring for the opioid analgesic activity of anilido-piperidines. The starting b-keto-amide 1.1, prepared by the aminolysis of methyl acetoacetate with methylphenethylamine, (93 % yield, was successively reacted with NaH and BuLi, to form the highly reactive a,g-dienolate anion 1.1a. Regio and chemoselective g-alkylation of the dienolate with various primary and secondary alkyl halides furnished the b-keto-amides 1.2–1.5 (76–91 %. Reductive amination of the keto-amides 1.1–1.5 with aniline and Zn powder in acetic acid, via the enamine intermediates 2.1–2.5, afforded the b-anilino amides 3.1–3.5 (74–85 %. After reductive deoxygenation of the tertiary amide group, using in situ generated diborane, the corresponding 1,3-diamines 4.1–4.5 were obtained (87–97 %. The synthesis of (±-2,3-seco-fentanyls 5.1–5.5 was completed by N-acylation of the diamines 4.1–4.5 with propionyl chloride, followed by precipitation of the monooxalate salts (86–95 %. The parent compound, 2,3-seco-fentanyl 5.1, was found to be a 40 times less potent narcotic analgesic than fentanyl but still 5–6 times more active than morphine in rats, while i-Pr derivative 5.3 was inactive. Apart from the pharmacological significance, the general procedure described herein may afford various functionalized, 1,3-diamines as potential complexing agents and building blocks for the synthesis of aza-crown ethers.

  15. CRITICAL POINTS IN THE FEEDING OF HIGH YIELDING DAIRY COWS IN ASSOCIATION WITH BCS AND METABOLIC PROFILE TEST

    Z. GERGÁCZ

    2013-07-01

    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

  16. Hibernation impact on the catalytic activities of the mitochondrial D-3-hydroxybutyrate dehydrogenase in liver and brain tissues of jerboa (Jaculus orientalis

    Hafiani Assia

    2003-09-01

    Full Text Available Abstract Background Jerboa (Jaculus orientalis is a deep hibernating rodent native to subdesert highlands. During hibernation, a high level of ketone bodies i.e. acetoacetate (AcAc and D-3-hydroxybutyrate (BOH are produced in liver, which are used in brain as energetic fuel. These compounds are bioconverted by mitochondrial D-3-hydroxybutyrate dehydrogenase (BDH E.C. 1.1.1.30. Here we report, the function and the expression of BDH in terms of catalytic activities, kinetic parameters, levels of protein and mRNA in both tissues i.e brain and liver, in relation to the hibernating process. Results We found that: 1/ In euthemic jerboa the specific activity in liver is 2.4- and 6.4- fold higher than in brain, respectively for AcAc reduction and for BOH oxidation. The same differences were found in the hibernation state. 2/ In euthermic jerboa, the Michaelis constants, KM BOH and KM NAD+ are different in liver and in brain while KM AcAc, KM NADH and the dissociation constants, KD NAD+and KD NADH are similar. 3/ During prehibernating state, as compared to euthermic state, the liver BDH activity is reduced by half, while kinetic constants are strongly increased except KD NAD+. 4/ During hibernating state, BDH activity is significantly enhanced, moreover, kinetic constants (KM and KD are strongly modified as compared to the euthermic state; i.e. KD NAD+ in liver and KM AcAc in brain decrease 5 and 3 times respectively, while KD NADH in brain strongly increases up to 5.6 fold. 5/ Both protein content and mRNA level of BDH remain unchanged during the cold adaptation process. Conclusions These results cumulatively explained and are consistent with the existence of two BDH enzymatic forms in the liver and the brain. The apoenzyme would be subjected to differential conformational folding depending on the hibernation state. This regulation could be a result of either post-translational modifications and/or a modification of the mitochondrial membrane state

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

    The thermal evolution of a sol–gel derived cobalt aluminate (CoAl2O4) of a spinel-type structure was studied. The samples were prepared by the sol–gel technique using aluminum-sec-butoxide [Al(OsBu)3] and cobalt nitrate hexahydrate Co(NO3)2 × 6H2O as starting materials and ethyl-acetoacetate (C6H10O3) as a chelating agent. The powder precursors were annealed at various temperatures ranging from 200 to 900 °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 °C a part of Co existed in a 3 + oxidation state. The accommodation of Co3+ in octahedral cation sites yields with a green color. Thermal treatment at higher temperatures promote the reduction of Co3+ ions to Co2+ and a change of color from green to blue, later being characteristic of Co2+ ions in tetrahedral coordination. The XRD revealed that annealing of the precursor powder at a lower temperature (500 and 600 °C) resulted in a crystal spinel-type phase Co2AlO4. Annealing at higher temperatures (700–900 °C) induces a reaction of Co2AlO4 with amorphous alumina producing a phase CoAl2O4 having a partially inverse spinel structure, with δ 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: ► Sol–gel derived spinel-type CoAl2O4 thermal evolution was monitored. ► Below 700 °C part of Co exists in 3+ state (octahedral sites), yielding green color. ► Above 700 °C Co3+ reduction to Co2+ (tetrahedral sites) is promoted, changing to blue. ► Treatment at 500–600 °C resulted in crystal spinel-type phase Co2AlO4. ► At 700–900 °C Co2AlO4 and amorphous alumina react

  18. 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: wolfgang.schaefer@uniklinik-freiburg.de [Department of Obstetrics and Gynecology, University Hospital Freiburg (Germany)

    2012-05-01

    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

  19. DIFFERENTIAL DIAGNOSIS OF ORGANIC ACIDEMIA: CLINICAL AND NEUROIMAGING FINDINGS

    Mahmoud Reza ASHRAFI

    2012-03-01

    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

  20. Chronic alcohol ingestion increases mortality and organ injury in a murine model of septic peritonitis.

    Benyam P Yoseph

    Full Text Available BACKGROUND: Patients admitted to the intensive care unit with alcohol use disorders have increased morbidity and mortality. The purpose of this study was to determine how chronic alcohol ingestion alters the host response to sepsis in mice. METHODS: Mice were randomized to receive either alcohol or water for 12 weeks and then subjected to cecal ligation and puncture. Mice were sacrificed 24 hours post-operatively or followed seven days for survival. RESULTS: Septic alcohol-fed mice had a significantly higher mortality than septic water-fed mice (74% vs. 41%, p = 0.01. This was associated with worsened gut integrity in alcohol-fed mice with elevated intestinal epithelial apoptosis, decreased crypt proliferation and shortened villus length. Further, alcohol-fed mice had higher intestinal permeability with decreased ZO-1 and occludin protein expression in the intestinal tight junction. The frequency of splenic and bone marrow CD4+ T cells was similar between groups; however, splenic CD4+ T cells in septic alcohol-fed mice had a marked increase in both TNF and IFN-γ production following ex vivo stimulation. Neither the frequency nor function of CD8+ T cells differed between alcohol-fed and water-fed septic mice. NK cells were decreased in both the spleen and bone marrow of alcohol-fed septic mice. Pulmonary myeloperoxidase levels and BAL levels of G-CSF and TFG-β were higher in alcohol-fed mice. Pancreatic metabolomics demonstrated increased acetate, adenosine, xanthine, acetoacetate, 3-hydroxybutyrate and betaine in alcohol-fed mice and decreased cytidine, uracil, fumarate, creatine phosphate, creatine, and choline. Serum and peritoneal cytokines were generally similar between alcohol-fed and water-fed mice, and there were no differences in bacteremia, lung wet to dry weight, or pulmonary, liver or splenic histology. CONCLUSIONS: When subjected to the same septic insult, mice with chronic alcohol ingestion have increased mortality

  1. Hyperglycemic hyperosmolar nonketotic syndrome.

    Venkatraman, R; Singhi, Sunit C

    2006-01-01

    Hyperglycemic hyperosmolar nonketotic syndrome (HHNS) was infrequently diagnosed till recently. Now it is being diagnosed with increasing frequency in obese children with type 2 diabetes mellitus (T2 DM) and its incidence is likely to go up, given global increase in incidence of childhood obesity, increased insulin resistance, and T2 DM. The syndrome is characterized by severe hyperglycemia, a marked increase in serum osmolality and dehydration without accumulation of beta -hydroxybutyric or acetoacetic ketoacids. Significant ketogenesis is restrained by the ability of the pancreas to secrete small amount of insulin. Prolonged phase of osmotic diuresis leads to severe depletion of body water, which excees that of sodium, resulting in hypertonic dehydration. These children, usually obese adolescents with T2 DM, present with signs of severe dehydration and depressed mental status but continue to have increased rather than decreased urine output and are at increased risk of developing rhabdomyolysis and malignant hyperthermia. Emergency treatment is directed at restoration of the intravascular volume, followed by correction of deficits of fluid and electrolyte (Na+, K+, Ca++, Mg++, PO4++), hyperglycemia and serum hyperosmolarity, and a thorough search for conditions that may lead to this metabolic decompensation and their treatment. Use of iso-osomolar isotonic fluid (0.9% saline) until hemodynamic stabilization initially, followed by 0.45% saline with insulin infusion at the rate of 0.1 units/kg/hour, addition of 5% dextrose in fluids and reduction of insulin infusion once the blood glucose is 250 to 300 mg/dl is generally recommended. However, evidence-based guidelines about composition and tonicity of fluids and electrolyte solutions for early resuscitation and rehydration, the rate of infusion-rapid vs slow, and insulin dose-low vs normal, in treatment of HHNS in children are awaited. Careful monitoring of glucose levels and ensuring adequate hydration in patients

  2. 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.

    2013-02-01

    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

  3. 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

    2016-08-01

    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

  4. 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

    2014-10-01

    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

  5. 1H nuclear magnetic resonance spectroscopy-based metabonomic study in patients with cirrhosis and hepatic encephalopathy

    Dabos, Konstantinos John; Parkinson, John Andrew; Sadler, Ian Howard; Plevris, John Nicholas; Hayes, Peter Clive

    2015-01-01

    AIM: To identify plasma metabolites used as biomarkers in order to distinguish cirrhotics from controls and encephalopathics. METHODS: A clinical study involving stable cirrhotic patients with and without overt hepatic encephalopathy was designed. A control group of healthy volunteers was used. Plasma from those patients was analysed using 1H - nuclear magnetic resonance spectroscopy. We used the Carr Purcell Meiboom Gill sequence to process the sample spectra at ambient probe temperature. We used a gated secondary irradiation field for water signal suppression. Samples were calibrated and referenced using the sodium trimethyl silyl propionate peak at 0.00 ppm. For each sample 128 transients (FID’s) were acquired into 32 K complex data points over a spectral width of 6 KHz. 30 degree pulses were applied with an acquisition time of 4.0 s in order to achieve better resolution, followed by a recovery delay of 12 s, to allow for complete relaxation and recovery of the magnetisation. A metabolic profile was created for stable cirrhotic patients without signs of overt hepatic encephalopathy and encephalopathic patients as well as healthy controls. Stepwise discriminant analysis was then used and discriminant factors were created to differentiate between the three groups. RESULTS: Eighteen stabled cirrhotic patients, eighteen patients with overt hepatic encephalopathy and seventeen healthy volunteers were recruited. Patients with cirrhosis had significantly impaired ketone body metabolism, urea synthesis and gluconeogenesis. This was demonstrated by higher concentrations of acetoacetate (0.23 ± 0.02 vs 0.05 ± 0.00, P < 0.01), and b-hydroxybutarate (0.58 ± 0.14 vs 0.08 ± 0.00, P < 0.01), lower concentrations of 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) and arginine (0.08 ± 0.01 vs 0.14 ± 0.02, P < 0.03) and higher concentrations of glutamate (1.36 ± 0.25 vs 0.58 ± 0.04, P < 0.01), lactate (1.53 ± 0

  6. 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

    2011-01-01

    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

  7. Adrenocortical and thyroid function, hormone and metabolite profiles and the onset of ovarian cyclicity in dairy cows suffering from various forms of ketosis

    Huszenicza Gyula

    2006-01-01

    Full Text Available The involvement of adrenocortical and thyroid hormones in the pathogenesis of ketosis, as well as the ovarian consequences of this metabolic disorder, were studied in _2 parity cows (n=199 in 3 large scale dairy herds. To compare the plasma/serum concentrations of certain hormones Šcortisol, thyroxin (T4, triiodothyronine (T3, insulin, insulin-like growth factor-1 (IGF-1¹ and metabolites Šglucose (G, acetoacetic acid (ACAC, βOH-butyrate (BHB, non-esterified fatty acid (NEFA, trigliceride (TG, total cholesterol (TCh¹, and the activity of aspartate aminotransferase (AST, blood samples were taken 1 to 3 days after calving and again 4 times 7 days apart. The ACTH-challenged cortisol responsiveness and the TRH-induced T4/T3 increase were determined between days 1 to 3 and again between days 28 to 35. The resumption of ovarian cyclicity was followed up by individual progesterone (P4 profiles based on milk samples taken 3 times a week for about 80 to 85 days. BHB level of 1 mmol/L was estimated as a border line between hyper- (>1 mmol/L and normoketonaemic (<1 mmol/L conditions. Five different ketone patterns were distinguished: (1 non-ketotic (n=98; normoketonaemia in all samples, (2 early type ketosis (n=45; hyperketonemia was detected only in the first week after calving, (3 late type (lactational ketosis (n=11; after a normoketonaemic period increasing hyperketonaemia was detected in the 5th, or in the 4th and 5th weeks, (4 temporary ketosis (n=11; hyperketonaemia was detected for 1-2 weeks in the 2nd and 3rd or in the 3rd and 4th weeks; (5 long-lasting ketosis (n=34; hyperketonaemia has been detected since calving for 4 to 5 weeks or until dying / emergency slaughtering. Simultaneously with the hyperketonaemic stage increased NEFA, ACAC, depressed TCh, glucose and decreased insulin, IGF-1, T4 and T3 concentrations were detected in almost all the cases. Obvious metabolic and endocrine alterations were found, however, only in long

  8. 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

    2007-04-01

    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

  9. Cloning, expression and characterization of a short-chain dehydrogenase from Pseudomonasfluorescens%荧光假单胞菌短链脱氢酶的克隆、表达及酶学性质分析

    薛群; 应向贤; 杨池; 汪钊

    2011-01-01

    为了研究荧光假单胞菌中短链脱氢酶的生理角色和催化特性,从荧光假单胞菌Pseudomonas fluorescens GIMl.49基因组DNA克隆表达了一个短链脱氢酶的编码基因pfd,并分析了该基因产物的酶学性质.基因pfd全长684 bp,编码227个氨基酸,推算分子量为24.2 kDa.将携带短链脱氢酶基因的重组质粒pET28b-pfd转入大肠杆菌BL21(DE3)进行表达,得到了28 kDa的表达产物.重组荧光假单胞菌短链脱氢酶(PFD)能氧化4-氯-3-羟基丁酸乙酯、1-苯乙醇、苯甲醇、仲丁醇和还原4-氯-乙酰乙酸乙酯、2-溴-苯乙酮、4-溴-苯乙酮等底物.以4-氯-3-羟基丁酸乙酯为底物时活力最高,Km值为186.40 mmol/L,Vmax为89.56 U/mg.氧化4-氯-3-羟基丁酸乙酯时,最适反应温度和pH分别为12℃和10.5,倾向于利用NAD+作辅酶;而还原4-氯-乙酰乙酸乙酯时,最适温度和pH为24℃和8.8,倾向于利用NADPH作辅酶.重组PFD能耐受50%(V/V)的甲醇等有机助溶剂,Ca2+(1 mmol/L)和EDTA(5 mmol/L)对其酶活有一定的促进作用.上述结果表明,重组PFD是一个新型的短链脱氢酶,其代谢角色推测与卤代次级醇的氧化降解有关.%To explore the physiological role and biocatalytic properties of short-chain dehydrogenases from Pseudomonas fluorescens GIM1.49, we cloned the structural gene pfd and characterized its over-expressed product. The length of gene pfd was 684 bp encoding a short-chain dehydrogenase with 227 amino acid residues and calculated molecular mass of 24.2 kDa. The recombinant plasmid pET28b-pfy was constructed and functionally expressed in Escherichia coli BL21(DE3), resulting in the over-production of recombinant short-chain dehydrogenase PFD with a size of 28 kDa. The enzyme could oxidize alcohols including 4-chloro-3-hydroxbutanoate ester and reduce 4-chloro-acetoacetate ester using either NAD(H) or NADP(H) as coenzyme. The enzyme showed the highest activity against 4-chloro-3-hydroxbutanoate ester as