Nitriles are an important type of synthetic intermediates in the production of fine chemicals because of their easy preparations and versatile transformations. The traditional chemical conversion of nitriles to carboxylic acids and amides is feasible but it requires relatively harsh conditions of heat, acid or alkali. Nitrile converting enzymes (nitrilase, nitrile hydratase and amidase) which are used as biocatalyst for the production of fine chemicals have attracted substantial interest because of their ability to convert readily available nitriles into the corresponding higher value amides or acids under mild conditions with excellent chemo-, regio- and stereo-selectivities. Many nitrile converting enzymes have been explored and widely used for the production of fine chemicals. In this paper, various examples of biocatalytic synthesis of pharmaceuticals and their intermediates, agrochemicals and their intermediates, food and feed additives, and other fine chemicals are presented. In the near future, an increasing number of novel nitrile converting enzymes will be screened and their potential in the production of useful fine chemicals will be further exploited.
Amides ; metabolism ; Amidohydrolases ; metabolism ; Aminohydrolases ; metabolism ; Carboxylic Acids ; metabolism ; Chemical Industry ; methods ; Hydro-Lyases ; metabolism ; Nitriles ; chemistry

OBJECTIVETo investigate the role of non-receptor tyrosine kinase Tec in the production of TNF-α and IL-1β from macrophages induced by LPS and its related mechanism.

METHODSRAW264.7 mononuclear-macrophages cultured in 6-well plates were divided into 4 groups according to the random number table, with 24 wells in each group. Cells in blank group were routinely cultured (cultured with DMEM medium containing 10% FBS) for 2 hours. Cells in LFM-A13 group were pretreated with 75 µmol/L Tec specific inhibitor LFM-A13 for 1 hour and then routinely cultured for 1 hour. Cells in LPS group were routinely cultured for 1 hour and then treated with 0.1 µg/mL LPS for 1 hour. Cells in LPS+LFM-A13 group were pretreated with 75 µmol/L LFM-A13 for 1 hour and then treated with 0.1 µg/mL LPS for 1 hour. The content of TNF-α and IL-1β in culture supernatant of cells was determined with ELISA. The mRNA expressions of TNF-α and IL-1β in cells were assayed with real-time fluorescent quantitative RT-PCR. The activity of intracellular Tec, p38 MAPK, and transforming growth factor activated kinase 1 (TAK1) was determined with Western blotting. Data were processed with one-way analysis of variance and LSD test.

RESULTSThe content of TNF-α and IL-1β in culture supernatant of cells in LFM-A13 group was close to that in blank group (with P values above 0.05). The mRNA expressions of TNF-α and IL-1β in the cells of LFM-A13 group were close to those of blank group (with P values above 0.05). The content of TNF-α and IL-1β in culture supernatant of cells in LPS group was respectively (1 213 ± 154) and (636 ± 90) pg/mL, which was higher than that in blank group [(330 ± 44) and (211 ± 31) pg/mL, with P values below 0.01]. The mRNA expressions of TNF-α and IL-1β in the cells of LPS group were respectively 1.57 ± 0.22 and 1.44 ± 0.24, which were significantly higher than those of blank group (1.00 ± 0.18 and 1.00 ± 0.19, with P values below 0.01). The content of TNF-α and IL-1β in culture supernatant of cells in LPS+LFM-A13 group was respectively (787 ± 109) and (453 ± 64) pg/mL, which was significantly lower than that in LPS group (with P values below 0.05). The mRNA expressions of TNF-α and IL-1β in the cells of LPS+LFM-A13 group were respectively 1.21 ± 0.15 and 1.21 ± 0.22, and they were significantly lower than those of LPS group (with P values below 0.05). The activity of intracellular Tec, TAK1, and p38 MAPK of cells in LPS+LFM-A13 group was close to that in blank group (with P values above 0.05). The activity of intracellular Tec, TAK1, and p38 MAPK of cells in LPS group was respectively 2.69 ± 0.41, 3.99 ± 0.65, and 2.07 ± 0.31, which was significantly higher than that in blank group (1.00 ± 0.17, 1.00 ± 0.16, and 1.00 ± 0.18, with P values below 0.01) and LPS+LFM-A13 group (1.02 ± 0.17, 1.18 ± 0.20, and 1.58 ± 0.28, P < 0.05 or P < 0.01).

CONCLUSIONSTec promotes the production and release of pro-inflammatory cytokines TNF-α and IL-1β from macrophages induced by LPS via TAK1-p38 MAPK signaling pathway.

Amides ; metabolism ; Cell Line ; Cytokines ; Interleukin-1beta ; metabolism ; secretion ; Lipopolysaccharides ; Macrophages ; metabolism ; Nitriles ; metabolism ; Protein-Tyrosine Kinases ; metabolism ; Signal Transduction ; Tumor Necrosis Factor-alpha ; metabolism ; secretion

To compare the cardiotoxicity induced by ropivacaine and bupivacaine and to investigate the mechanism of cardiotoxicity, 24 mature New Zealand rabbits were divided randomly into control group (group C), ropivacaine group (group R) and bupivacaine group (group B). Hearts were drawn out rapidly from the anesthetized animals and cardiac perfusion was performed immediately. Ropivacaine 500 ng/ml (group R) or bupivacaine 500 ng/ml (group B) was added to the perfusion solution. Ventricular myocardial ATP, ADP and AMP were measured with high performance liquid chromatogram. The ability of myocardial mitochondria oxidation to pyruvate or palmitoylcarnitine was detected with Clark electrode. Our results showed that myocardial ATP and ADP decreased significantly (P < 0.05) in group R and most significantly (P < 0.01) in group B as compared with group C. Myocardial ATP and ADP decreased most significantly (P < 0.01) in group B as compared with group R. The changes of myocardial AMP revealed significant difference among three groups. The changes of pyruvate oxidation exhibited no significant difference among the three groups. Palmitoylcarnitine oxidation decreased markedly (P < 0.05) in group R and most significantly (P < 0.01) in group B as compared with group C. The present study indicated that the inhibition of lipid substrate oxidation may be responsible for the cardiotoxicity induced by bupivacaine and ropivacaine. The cardiotoxicity induced by ropivacaine is far more less than bupivacaine.
Adenosine Diphosphate ; metabolism ; Adenosine Triphosphate ; metabolism ; Amides ; toxicity ; Anesthetics, Local ; toxicity ; Animals ; Bupivacaine ; toxicity ; Energy Metabolism ; In Vitro Techniques ; Mitochondria, Heart ; metabolism ; Myocardium ; metabolism ; Oxidation-Reduction ; Rabbits ; Random Allocation

BACKGROUNDGastric cancer (GC) is one of the most common types of cancer in the world. A change in the metabolism of lipids in tumor cells could lead to the pathogenesis of cancer. In this study, we investigated fatty acid and fatty acid amide metabolic perturbations associated with GC morbidity.

METHODSGas chromatography/mass spectrometry (GC/MS) was utilized to analyze fatty acids (FAs) and fatty acid amides (FAAs) of GC tissues and matched normal mucosae from 30 GC patients. Acquired lipid data was analyzed using non parametric Wilcoxon rank sum test to find the differential biomarkers for GC and diagnostic models for GC were established by using orthogonal partial least squares discriminant analysis (OPLS-DA).

RESULTSA total of 13 FAs and 4 FAAs were detected using GC/MS and 5 differential FAs as well as oleamide were identified with significant difference (P<0.05). The OPLS-DA model generated from lipid profile showed adequate discrimination of GC tissues from normal mucosae while the OPLS-DA model failed to separate GC specimens of different TNM stages. A total of 8 variables were obtained for their most contribution in the discriminating model (Variable importance in the projection (VIP) value>1.0), five of which were detected with significant difference (P<0.05).

CONCLUSIONSFA and FAA metabolic profiles have great potential in detecting GC and helping understand perturbations of lipid metabolism associated with GC morbidity.

Amides ; metabolism ; Fatty Acids ; metabolism ; Female ; Gas Chromatography-Mass Spectrometry ; Humans ; In Vitro Techniques ; Male ; Metabolic Diseases ; physiopathology ; Stomach Neoplasms ; metabolism ; pathology

OBJECTIVE[corrected] To assess the effects of metallothionein on myocyte apoptosis and energy supply of isolated rabbit heart muscle during perfusion with ropivacaine..

METHODSSixty New Zealand white male rabbits were randomized into 3 equal groups. In group I, the rabbits received a intreaperitioneal injection of distilled water 24 h before isolation of the heart with perfusion by Langendoff model; in group II, distilled water was injected intreaperitioneally, and 24 h later the heart was isolated and perfused with Langendoff model and ropivacaine; in group III, 3.6% ZnSO(4) was injected intreaperitioneally and the isolated heart was perfused with Langendoff model and ropivacaine. The myocardial metallothionein content, myocyte apoptosis, and myocardial ATP, ADP and AMP content were detected.

RESULTSThe myocardial metallothionein content was significantly higher in group III than in the other two groups; the percent of myocyte apoptosis was the highest in group II, and was significantly higher in group III than in group I. The myocardial content of ATP was the highest in group I, and was significantly higher in group III than in group II.

CONCLUSIONMetallothionein can significantly inhibit myocyte apoptosis and alleviate energy supply disorder induced by ropivacaine.

Amides ; pharmacology ; Animals ; Apoptosis ; drug effects ; Energy Metabolism ; drug effects ; In Vitro Techniques ; Male ; Metallothionein ; pharmacology ; Myocardium ; cytology ; metabolism ; Myocytes, Cardiac ; cytology ; metabolism ; Perfusion ; Rabbits

Ten known phenolic compounds including [4]-gingerol (1), [6]-gingerol (2), [10]-gingerol (3), (3S,5S)-3,5-dihydroxy-1-(4-hydroxy-3-methoxyphenyl) decane (4), (3R,5S) -3, 5-dihydroxy-1-(4-hydroxy-3-methoxyphenyl) decane (5), [6]-shogaol (6), [10]-shogaol (7), gingerenone A (8), hexahydrocurcumin (9), and (3R,5R)-3,5-dihydroxy-1,7-bis(4-hydroxy-3-methoxyphenyl) heptane (10), and seven amides including piperine (11), isochavicine (12), isopiperine (13), N-trans-p-coumaroyl tyramine (14), N-trans-feruloyl tyramine (15), N-trans-p-coumaroyl octopamine (16), N-trans-feruloyl octopamine (17), were isolated and identified from the roots of Lycianthes marlipoensis. Compounds 1-13 and 17 were isolated from the genus Lycianthes for the first time.
Amides ; chemistry ; isolation & purification ; Chromatography ; methods ; Magnetic Resonance Spectroscopy ; methods ; Phenols ; chemistry ; isolation & purification ; Plant Extracts ; chemistry ; isolation & purification ; Plant Roots ; chemistry ; metabolism ; Solanaceae ; chemistry ; metabolism

AIMTo explore the synthetic methods of polypeptides containing new heart of kidney imaging agents.

METHODS AND RESULTSFive new target chelators--2-N-(2'-s-triphenylmethylacetyl) amino-(N'-2"-N",N"-diethylethylamine) phenylpropamide (MPNE), 2-N-(2'-s-triphenylmethyl acetyl) amino-(N'-2"-N",N"-dimethylethylamine) phenylpropamide (MPNM), 2-N-(2's-triphenylmethylacetyl) amino-3-methyl-(N'-2"-N",N"-dimethylethylamine) butyramide (MVNM), 2-N-(2'-s-triphenyl methylacetyl) amino-3-methyl-(N'-2"-N",N"-diethylethylamine) butyramide (MVNE), 2-N-(2'-s-triphenylmethylacetyl) amino-(N'-acetylglycine) phenylpropamide (MPG2)--were synthesized through five steps with mercaptoacetic acid as primitive materials, all of which were identified on the basis of spectroscopic data, such as IR, 1HNMR, MS or elementary analysis. The protection of the mercapto group was improved and the relatively new reaction condition of active ester with amino acid is developed. All the chelators were labeled with Technetium-99m and their biological activities in mice given in ID values was tested to explore new heart imaging agents, where ID is the percentage injected dose per organ. The ID was determined by in vivo biodistribution study. Tc-99m complexes 0.1 mL was injected into the laterial tail vein of 3 anaesthetised rats. At 2, 5, 10, 30, 60 min post-injection, rats were sacrificed by decapitation, bled from the neck and dissected. Organs were removed at dissection. The radioactivities in various organs were determined in an automatic twin crystal gamma counter.

CONCLUSIONThe bio-distribution results in mice indicate that 99Tcm-MVNM have higher heart uptake (ID = 8.40%/g, 2 min post-injection) and quicker blood clearance (ID = 4.3%/g, 60 min post-injection); 99Tcm-MPNE and 99Tcm-MPNM also have fairly high heart uptake and quick blood clearance; 99Tcm-MPG2 has better kidney accumulation and higher activity ratios of kidney to blood (about 4).

Amides ; chemical synthesis ; pharmacokinetics ; Animals ; Kidney ; metabolism ; Mice ; Molecular Structure ; Myocardium ; metabolism ; Organotechnetium Compounds ; chemical synthesis ; pharmacokinetics ; Peptides ; chemical synthesis ; chemistry ; pharmacokinetics ; Sulfides ; chemical synthesis ; pharmacokinetics ; Tissue Distribution

OBJECTIVE: To observe the behaviour and Ca2+ content of spinal cord in rats after continuous spinal anesthesia administration of different densities and doses of ropivacaine in SD rats. METHODS: Twenty-four male SD rats weighing 220 approximately 280 g were anesthetized. A polyurethane microspinal catheter was inserted into the lumbar subarachnoid space 8 cm according to the method of Yaksh's. The animals were randomly divided into 4 groups of 6 each: in group N the rats were given normal saline 40 microl intrathecally every one and half hours for 3 times; in group R1, 0.5% ropivacaine was given; in group R2 0.75% ropivacaine and in group R3 1% ropivacaine was given. The activity of rats was observed. After 6 hours rats were perfused with 4% formamint through the ascending aorta. The rats were sacrificed and L1 approximately 2 segment of spinal cord was immediately removed for Ca2+ content examination. RESULTS: A total hind limbs paralysis was seen at 30 seconds and intramuscular strain gradually came back from 30 to 90 minutes after the intrathecal administration of ropivacaine in all rats. The recovery of motor black was remarkably different in group R1, R2, and R3 (P < 0.05). The Ca2+ content of spinal cord was significantly higher in group R3 than that in group R1 and R2 (P < 0.05 ). CONCLUSION There is no significant change of motor black time and it is related to drug dose for 0.5% , 0.75% and 1% ropivacaine in continuous spinal anesthesia. 1% ropivacaine may increase Ca2+ content in spinal cord.
Amides ; pharmacology ; Anesthesia, Spinal ; Animals ; Behavior, Animal ; Calcium ; metabolism ; Dose-Response Relationship, Drug ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Ropivacaine ; Spinal Cord ; metabolism

OBJECTIVE: To explore the relationship of changes of amide A in rabbit heart and the postmortem interval (PMI) by FTIR spectroscopy technique. METHODS: Thirty-two rabbits were sacrificed and the hearts were sampled at 20 degrees C within 48 h postmortem points. All samples were sliced and tested by FTIR spectroscopy technique. The images of amide A were created by FTIR spectroscopic imaging. The positive and negative area ratios of amide A were analyzed using imaging analysis system. RESULTS: The positive and negative area ratios declined regularly with the prolongation of death time in 48 h. There was a significant quadric relationship between the area ratios (y) of amide A (positive and negative area) and PMI(x). The regression equation was y = 0.001x2-0.038x + 0.747(R2 = 0.940). CONCLUSION The ratios of positive and negative area of amide A showed a strong correlation with PMI and could be used to estimate PMI.
Amides/metabolism* ; Animals ; Female ; Forensic Pathology/methods* ; Male ; Myocardium/metabolism* ; Postmortem Changes ; Rabbits ; Regression Analysis ; Spectroscopy, Fourier Transform Infrared ; Time Factors

OBJECTIVETo investigate the changes in endothelial cytoskeletal reorganization and the role of Rho in the signal transduction pathway.

METHODSECV304 cells were cultured and randomly divided into following groups: i.e. sham (with normal rat serum treatment), burn (with burn rat serum treatment), Y (with 30 micromol/L Rho kinase inhibitor Y-27632 treatment), burn plus Y (pretreatment of cells with burn serum before treated with 30 micromol/L Y-27632), Y plus burn (pretreatment of cells with Y-27632 for 1 hour before treated with burn serum), LPA (with normal rat serum and 13 micromol/L LPA), and LPA plus Y (pretreatment of cells with LPA before treated with Y-27632) groups. The indices were examined at 6, 7 and 8 posttreatment hours (PTH) in all groups except in Y group. The endothelial morphology was observed with HE staining. Endothelial cytoskeleton was observed by dual-fluorescence labeling of filamenta (F) with Rhodamine-phalloidin and monomer (G) with oregon green labeled DNAase. The actin content in the cells in all groups was measured with flow cytometry.

RESULTSIn sham and control group, the cells were in fusiform or polygonal shape, with satisfactory growth; filamentous actin (F-actin) was mainly distributed in the peripheral site of the cytoplasm and formed peripheral filamental band. The cells became confluent to form a single layer with reticular structure. Globular actin (G-actin) was concentrated in the nucleus and per nucleus. In burn group, after 6 hours of burn serum treatment, the ability of cells to adhere to vessel wall was weakened, and a striking reorganization of the actin cytoskeleton and the formation of the stress fibers were found. Furthermore, the fluorescent intensity of the peripheral filament bands was weakened, and dispersed actin monomers were seen in the cytoplasm. This reaction was enhanced along with elapse of stimulation time. In burn plus Y or Y plus burn group, the cells grew and adhered well to the wall of culture vessel. The distribution of the filamentous actin was the same as the sham group, while the stress fiber decreased in amount obviously. The structure of globular actin was condensed with little G-actin in the cytoplasm. The changes in actin cytoskeleton in LPA group was similar to that in burn group. The effects of LPA on actin reorganization could also be reversed by Y-27632. The content of F-actin in burn group at 6 PTH (0.63 +/- 0.07) was lower than that in sham group (0.75 +/- 0.08), while the content of G-actin in burn group (1.28 +/- 0.27) was higher than that in sham group (1.16 +/- 0.16, P > 0.05).

CONCLUSIONBurn serum induces vascular endothelial actin cytoskeleton reorganization in endothelial cells via the Rho-dependent signal pathway. Similar to the effect of LPA, this effect could be reversed by Y-27632.

Actins ; metabolism ; Amides ; pharmacology ; Animals ; Burns ; blood ; metabolism ; Cells, Cultured ; Cytoskeleton ; metabolism ; Endothelial Cells ; metabolism ; Endothelium, Vascular ; Humans ; Male ; Pyridines ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Serum ; metabolism ; Signal Transduction ; rho-Associated Kinases ; antagonists & inhibitors ; metabolism