共查询到20条相似文献,搜索用时 46 毫秒
1.
M. Ardhaoui A. Falcimaigne J. M. Engasser P. Moussou G. Pauly M. Ghoul 《Biocatalysis and Biotransformation》2004,22(4):253-259
Enzymatic acylation of rutin and esculin with aromatic, aliphatic and aryl-aliphatic acids using Candida antarctica lipase in tert amyl alcohol as solvent was investigated under low water content. Whatever the acyl donor used, the conversion yields and initial rates for esculin were higher than for rutin. For a given flavonoid, the performance of these reactions depended on the acyl donor structures. For aliphatic acids, conversion yields and initial rates of both flavonoids were respectively in the ranges of 68-90% and of 9.5×10-2-72×10-2 mmol l-1 h-1. For aromatic acids, the reaction occurred only with the aryl subgroup (cinnamic, hydrocinnamic, 3,4-dihydroxyhydrocinnamic and 4-hydroxyphenyl acetic acids) and was drastically influenced by the presence of side chain and substitution patterns of the aromatic ring. Except for hydrocinnamic acid (75%, 23.4×10-2 mmol l-1 h-1), with these acids the conversion yields and initial rates were lower and in the range of 10-45% and of 0.7×10-2 to 12.1×10-2 mmol l-1 h-1. Unsaturation of the side chain of the hydrocinnamic acid decreased the esculin conversion rate from 75 to 13% and initial rate from 23.4 to 1.76×10-2 mmol l-1 h-1. The presence of hydroxyl or nitro-groups on the aromatic ring of the aryl aliphatic acid also reduced conversion yields and initial rates. Even without a spacer, the non-phenolic ring acid (quinic acid) was reactive and lead to conversion yields of about 20 and 23% respectively for rutin and esculin. 相似文献
2.
Noppol Leksawasdi Michael Breuer Bernhard Hauer Bettina Rosche Peter L. Rogers 《Biocatalysis and Biotransformation》2003,21(6):315-320
Based on experimental data, a kinetic model for the deactivation of partially purified pyruvate decarboxylase (PDC) by benzaldehyde (0-200 mM) in MOPS buffer (2.5 M) has been developed. An initial lag period prior to deactivation was found to occur. With first order dependencies of PDC deactivation on exposure time and on benzaldehyde concentration, a reaction time deactivation constant of 2.64×10-3 h-1 and a benzaldehyde deactivation coefficient of 1.98×10-4 mM-1 h-1 were determined for benzaldehyde concentrations up to 200 mM. The PDC deactivation kinetic equations established in this study are an essential component in an overall model being developed to describe the enzymatic biotransformation of benzaldehyde and pyruvate to produce the pharmaceutical intermediate (R)-phenylacetylcarbinol (R-PAC). 相似文献
3.
A set of conditions was developed for the specific binding of acriflavine to the DNA of intact squamous cells. This was achieved through a series of studies into the relative affinities for dye between DNA and various biopolymers by an agar gel diffusion technique. Specificity was ascertained by DNase and RNase treatment of the cells. The final conditions, based on an estimated DNA-to-dye ratio of 4:1, required a constant cell count of 100,000 and dye at a concentration of 0.0025 μg per ml in 10 ml of phosphate buffer, pH 6.0-7.4. These quantities were dictated by the sensitivity limitations of the analytical apparatus. To make use of standard fluorometric instrumentation, the whole cell population method for determining average values was followed. Free dye was analyzed after cell samples were stained, and the difference between this value and that of an aliquot of working dye was taken as the amount of bound dye. To ensure cell-free residues of dye, cells were removed by centrifugation through Teflon membrane filters. The average amount of dye which was bound to the DNA of normal squamous cells was 1.6 × 10-7 μg (0.7 × 10-15 mole) per cell, and the amount bound to HeLa cells was 2.3 × 10-7 μg (1 × 10-15 mole). These values were highly replicable, making it possible to use them as an expression of the DNA content of the cells. The mildness of the staining conditions, and the preservation of cellular integrity in this technique may permit quantitative measurements of the DNA content of living cells. 相似文献
4.
M. G. Roig F. J. Burguillo N. I. Ghais B. Velasco J. M. Cachaza 《Biocatalysis and Biotransformation》1993,7(2):97-115
The kinetic behavior of human placental alkaline phosphatase, which catalyses the hydrolysis of p-nitrophenyl and of o-carboxyphenyl phosphates, was studied by means of graphical and non-linear regression statistical fitting analysis of data of rate versus substrate concentration. Non linear Lineweaver-Burk and Eadie-Hofstee plots and rational functions of degree 2:2 (F-test assessing the goodness of fit) show non-Michaelian kinetic behavior. In the same way, the behavior of the enzyme was also non-Michaelian in the simultaneous presence of these two substrates.
Norlaudanosoline is a key intermediate in the biosynthesis of the benzylisoquinoline alkaloids providing the benzyl-isoquinoline portion of the morphinan skeleton. This study examines a coupled reaction system for the production of norlaudanosoline from dopamine. In this coupled system, dopamine is enzymatically converted by monoamine oxidase (MAO) to 3,4-dihydroxyphenylacetaldehyde (dopaldehyde). In the presence of dopamine, this aldehyde undergoes a spontaneous Pictet-Spengler condensation to form norlaudanosoline. Three potential sources of MAO were investigated: a fungal source (Aspergillus niger), a bacterial source (Sarcina lutea) and a commercial source isolated from bovine plasma. Kinetic studies with dopamine as the substrate gave Michaelis constants (Km) of 1.81 × 10-5 M, 6.94 × 10-3, and 1.61 × 10-3 M for A. niger, S. lutea and bovine plasma oxidase, respectively. The reaction system is complicated because of the effect of the condensation reaction, so a more rigorous model was developed to account for this effect. The model was suitable for showing the effect of dopamine concentration on norlaudanosoline production alghough there were some model inadequacies. Using the model a forward rate constant for the Pictet-Spengler condensation was determined to be 6.8 × 10-2 M-1 s-1 and the reverse reaction appears to be negligible. Overall conversion was 14% which is 20 times that achieved in an in situ reaction system using whole cells of Aspergillus niger. 相似文献
Norlaudanosoline is a key intermediate in the biosynthesis of the benzylisoquinoline alkaloids providing the benzyl-isoquinoline portion of the morphinan skeleton. This study examines a coupled reaction system for the production of norlaudanosoline from dopamine. In this coupled system, dopamine is enzymatically converted by monoamine oxidase (MAO) to 3,4-dihydroxyphenylacetaldehyde (dopaldehyde). In the presence of dopamine, this aldehyde undergoes a spontaneous Pictet-Spengler condensation to form norlaudanosoline. Three potential sources of MAO were investigated: a fungal source (Aspergillus niger), a bacterial source (Sarcina lutea) and a commercial source isolated from bovine plasma. Kinetic studies with dopamine as the substrate gave Michaelis constants (Km) of 1.81 × 10-5 M, 6.94 × 10-3, and 1.61 × 10-3 M for A. niger, S. lutea and bovine plasma oxidase, respectively. The reaction system is complicated because of the effect of the condensation reaction, so a more rigorous model was developed to account for this effect. The model was suitable for showing the effect of dopamine concentration on norlaudanosoline production alghough there were some model inadequacies. Using the model a forward rate constant for the Pictet-Spengler condensation was determined to be 6.8 × 10-2 M-1 s-1 and the reverse reaction appears to be negligible. Overall conversion was 14% which is 20 times that achieved in an in situ reaction system using whole cells of Aspergillus niger. 相似文献
5.
S. E. Unkles C. Marriott J. R. Kinghorn C. Panter A. Blackwell 《Biocontrol Science and Technology》2004,14(4):397-401
Culicinomyces clavisporus, a fungal pathogen of a wide range of mosquito species, was investigated in relation to potential pathogenicity against Culicoides nubeculosus biting midge larvae. Seven different C. clavisporus strains were assayed. Each showed some degree of activity against C. nubeculosus larvae with LC50 values of between 3.2×10-5 and 1.1×10-6 spores/mL; these effects occurred in dose-dependent manners and tended to be delayed until 72-96 h post treatment. The results are discussed in relation to incorporation of C. clavisporus into biocontrol programmes for Culicoides spp. 相似文献
6.
Mechanism of reaction of nitrogen dioxide radical with hydroxycinnamic acid derivatives: A pulse radiolysis study 总被引:5,自引:0,他引:5
Zhang Zhouen Yao Side Lin Weizhen Wang Wenfeng Jin Yizun Lin Nianyun 《Free radical research》1998,29(1):13-16
Nitrogen dioxide radical (NO·2) is known as a toxic agent produced in the metabolism of nitrates and nitrites. By the use of the pulse radiolysis technique, the mechanism of the reaction of NO·2 radical with hydroxycinnamic acid derivatives (HCA) was studied and the rate constants have been measured. The rate constants were found to be 7.4 × 108, 7.2 × 108, 8.6 × 108 dm3 mol-1s-1 for ferulic acid, sinapic acid and caffeic acid, respectively. The reactions produce the corresponding phenoxyl radical. 相似文献
7.
Amada Alvarez Reimer 《Journal of experimental marine biology and ecology》1972,10(3):177-181
Palythoa psammophilia Walsh & Bowers has a well coordinated, stereotyped feeding response, the culminating step of which is ingestion; this may be elicited by the synergistic effect of the tripeptide glutathione and the -imino acid, proline. Either activator acting separately causes responses only at high concentrations (above 10−5 M for glutathione; above 10−4 M for proline) in a reduced number of animals and at a low rate (5.00 ± 1.73 min in 5 × 10−3 M solutions of glutathione; 11.10±3.74 min in 5 × 10−3 M solutions of proline). Highest percentages of response were obtained in combinations where glutathione was at a concentration of 5 × 10−3 M and proline at 5 × 10−4 M or in combinations of glutathione at concentrations 5 × 10−6 M and proline at 5 × 10−5 M. The speed of ingestion is considerably enhanced when these activators are combined (1.17±1.18 min). 相似文献
8.
Peeyoosh Kant Pandey Arti Srivastava Jasaswini Tripathy Kunj Behari 《Carbohydrate polymers》2006,65(4):414-420
Guar gum has been modified by graft copolymerization with acrylic acid in aqueous medium using vanadium (V)–mercaptosuccinic acid redox system. The optimum reaction conditions affording maximum grafting ratio, efficiency, add on and conversion have been determined. The grafting parameters have been found to increase with increase in vanadium (V) concentration upto 1.0 × 10−2 mol dm−3, but these parameters decrease on further increasing the vanadium (V) concentration. On increasing the mercaptosuccinic acid concentration from 1.0 × 10−2 to 4.0 × 10−2 mol dm−3 grafting ratio, efficiency and add on increase up to 2.0 × 10−2 mol dm−3 but decrease with further increase in mercaptosuccinic acid concentration. On varying the acrylic acid concentration from 5.0 × 10−2 to 30.0 × 10−2 mol dm−3, maximum grafting ratio, efficiency and add on have been obtained at 20.0 × 10−2 mol dm−3. The grafting ratio, add on and conversion increase, on increasing the H+ ion concentration from 1.5 × 10−1 to 6.0 × 10−1 mol dm−3. On increasing the guar gum concentration the grafting parameters increase. The grafting ratio, add on and conversion have been found to increase with time period while efficiency started decreasing after 120 min. It has been observed that %G increases on increasing the temperature up to 35 °C. The graft copolymer has been characterized by IR spectroscopy and thermogravimetric analysis. 相似文献
9.
The electron transfer reactions between the trichloromethylperoxyl radical (Cl3COO·) and hydroxycinnamic acid derivatives, including chlorogenic acid, sinapic acid, caffeic acid, ferulic acid and 3,4-(methylenedioxy)cinnamic acid, have been studied by pulse radiolysis. The hydroxycinnamic acid derivatives, especially sinapic acid, are identified as good antioxidants for reduction of Cl3COO· via electron transfer reactions. From buildup kinetic analysis of phenoxyl radical, the rate constant for reaction of Cl3COO· with sinapic acid has been determined to be 8.2 × 107 dm3 mol-1 s-1, while the rate constants of electron transfer from other hydroxycinnamic acid derivatives to Cl3COO· were obtained to be about 2 × 107dm3 mol-1 s-1. The reaction of 3,4-(methylenedioxy)cinnamic acid with Cl3COO· was investigated as an evidence for the electron transfer mechanism. 相似文献
10.
A. Curulli S. Kelly C. O'Sullivan G. G. Guilbault G. Palleschi 《Biosensors & bioelectronics》1998,13(12):1245-1250
An electrochemical biosensor for the determination of lysine to be used for rapid evaluation of food quality has been developed. Platinum electrodes have been coated by electropolymerisation with 1,2-diaminobenzene (1.2-DAB) using cyclic voltammetry. The reduction in the oxidation of interferents compared with the bare platinum electrode was 100% for ascorbic acid, 99% for acetaminophen and 99% for cysteine. The enzyme L-lysine--oxidase was then immobilised onto the polymer layer by passive adsorption and a calibration curve for lysine constructed. This gave a linear range of 1×10−5 mol/l to 1×10−3 mol/l and a limit of detection of 2×10−7 mol/l. 相似文献
11.
David B. Green Stephen M. Dodge Jeffrey R. Lee Gary Tallman 《Biotechnic & histochemistry》1990,65(1):15-24
The effect was examined of the chemical decomposition of the potassium stain, sodium hexanitrocobaltate (III) (SHC), on its ability to produce stain granules of consistent size that could be used to estimate the K+ contents of stomafal guard cells. Stomata in detached epidermis from leaves of Vicia faba (fava bean) were stimulated to accumulate K+ by treating them with fusicoccin. Stomatal apertures and the fraction of guard cell area covered by K+ precipitate granules (K+ score) were measured by digitizing photographic enlargements, and K* scores were correlated with the age of stain that had been stored either in open or closed containers. The ability of stain aged in open containers to produce consistent fractional cell coverage was compared to 1) the ability of identically treated stain to precipitate K+ from solutions of KCl, and to 2) the kinetics of decomposition of SHC. It was found that the fractional coverage of guard cells of stomata opened to the same apertures decreased with a first order rate constant of 2.3 × 10-8/sec. The mass of precipitate formed by treatment of KCl solutions was unchanged for 2 hr after initial preparation of the SHC, and decreased thereafter with a first order rate constant of 1.0 × 10-5/sec. When stored in tightly sealed containers, nearly 100 hr were required for an occasionally opened bottle of SHC to decay to the same efficacy as a solution left open to the air for 8 hr. 相似文献
12.
Nicolas Van Larebeke Gudrun Koppen Vera Nelen Greet Schoeters Herman Van Loon Harma Albering Louk Riga Robert Vlietinck Jos Kleinjans 《Biomarkers》2004,9(1):71-84
Biomarkers were measured in residents of Wilrijk and Hoboken, industrial suburbs of the city of Antwerp, and of Peer, a rural municipality in Flanders, Belgium. Persons with known occupational exposures to toxic compounds or commuting over long distances were excluded. Here, we report the hypoxanthine phosphoribosyltransferase gene (HPRT) variant frequencies for 99 non-smoking women aged 50-65 years. HPRT values above the detection limit (Vfpos values) were observed for 43 subjects (21 from Peer, 22 from Antwerp). The median (10th to 90th percentiles) HPRT variant frequency (Vfpos) in peripheral lymphocytes was 9.59 (3.44-56.99) for Peer and 3.57 (1.57-13.96) for Antwerp. The Vfpos value was significantly higher in Peer than in Antwerp, both in terms of crude data (p=0.011) and after correction for age, level of education, smoking status, serum level of selenium and body mass index through analysis of covariance (p=0.011). For the total study population, serum lead concentration showed a non-significant positive correlation with lnVfpos. In addition, subjects with a blood lead concentration above the median tended to have higher Vfpos values (9.45×10-6 for 'high' group versus 5.21×10-6 for 'low' group; p=0.077 after correction for confounding). Subjects with a serum selenium level above the median tended to have lower Vfpos values (4.99×10-6 for 'high' group versus 9.83×10-6 for 'low' group; p=0.051 after correction for confounding). These data are consistent with an indirect genotoxic effect of lead and with an antimutagenic effect of selenium. 相似文献
13.
Effect of reaction temperature and reaction time on the preparation of low-molecular-weight chitosan using phosphoric acid 总被引:6,自引:0,他引:6
Low-molecular-weight chitosan were prepared using 85% phosphoric acid at different reaction temperatures and reaction time. At room temperature, the viscosity average-molecular weights (Mv) of chitosan decreased to 7.1×104 from 21.4×104 after 35 days treatment. The degradation rate decreased with increasing hydrolysis time. The yields of chitosan also continuously decreased from 68.4 to 40.2% after 35 days. At 40, 60 and 80 °C, the molecular weight decreased to 3.70×104, 3.50×104 and 2.00×104 on 8 h hydrolysis, respectively. The yields of chitosan remain at a high level compared with that at room temperature and were 86.5, 71.4 and 61.3% at 40, 60 and 80 °C treatment, respectively. The different reaction time gave chitosan with different molecular weights. At 60 °C, the molecular weight of products decreased to 7.40×104 from 21.4×104 within 4 h, then decreased slowly to 1.90×104 in 15 h. It was also found that the water-solubility of chitosan increased as the molecular weight decreased. Results show the changes in yields and molecular weight of chitooligomers were strongly dependent on the reaction temperature and reaction time. 相似文献
14.
A plant tissue biosensor associated with flow injection analysis is proposed to determine epinephrine in pharmaceutical samples. The polyphenol oxidase enzymes present in the fibers of a palm tree fruits (Livistona chinensis), catalyses the oxidation of epinephrine to epinephrinequinone as a primary product. This product is then electrochemically reduced (at −0.10 V versus Ag/AgClsat) on the biosensor surface and the resulting current is used for the quantification of epinephrine. The biosensor provides a linear response for epinephrine in the concentration range from 5.0 × 10−5 to 3.5 × 10−4 mol l−1. The limit of detection estimated for this interval was 1.5 × 10−5 mol l−1 and the correlation coefficient of 0.998, working under a flow rate of 2.0 ml min−1 and using a sample loop of 100 μl. The repeatability (R.S.D. for 10 consecutive determinations of a 3.0 × 10−4 mol l−1 epinephrine solution) was 3.1%. The results obtained by the method here proposed were compared with the official UV spectrophotometric procedure and also using a plant tissue reactor. The responses obtained with the proposed strategies were in good agreement with both ways of analyses, whereas the values obtained by the official spectrophotometric method was strongly affected by benzoic acid, present in the formulation of pharmaceutical product utilized for inhalation. Such favorable results obtained with the carbon paste biosensor or utilizing the bioreactor, joined with the simplicity of its preparation turns these procedures very attractive for epinephrine quantification in pharmaceutical products. 相似文献
15.
The free radical (II) produced by one-electron reduction of adriamycin (I) exists in aqueous solution at pH 7.0 in equilibrium with the parent and the two-electron reduced form (III). Over some hundreds of milliseconds deglycosylation takes place yielding an aglycone (IV) which subsequently rearranges to form a more stable aglycone. 7-deoxyadriamycinone (V). The changes in the optical absorption spectrum accompanying these processes are reported. The rate constant for III + IV is 1.1 s-1 and for IV + V is 1.5 × 10--2 s.-1. At pH 4.0 the two electron reduced form of adriamycin exists predominantly in a different tautomeric form (VII). It is suggested that this deglycosylates via a free radical mechanism involving the acidic form of the semiquinone free radical (VI) 相似文献
16.
Amperometric choline biosensors were fabricated by the covalent immobilization of an enzyme of choline oxidase (ChO) and a bi-enzyme of ChO/horseradish peroxidase (ChO/HRP) onto poly-5,2′:5′,2″-terthiophene-3′-carboxylic acid (poly-TTCA) modified electrodes (CPMEs). A sensor modified with ChO utilized the oxidation process of enzymatically generated H2O2 in a choline solution at +0.6 V. The other one modified with ChO/HRP utilized the reduction process of H2O2 in a choline solution at −0.2 V. Experimental parameters affecting the sensitivity of sensors, such as pH, applied potential, and temperature were optimized. A performance comparison of two sensors showed that one based on ChO/HRP/CPME had a linear range from 1.0×10−6 to 8.0×10−5 M and the other based on ChO/CPME from 1.0×10−6 to 5.0×10−5 M. The detection limits for choline employing ChO/HRP/CPME and ChO/CPME were determined to be about 1.0×10−7 and 4.0×10−7 M, respectively. The response time of sensors was less than 5 s. Sensors showed good selectivity to interfering species. The long-term storage stability of the sensor based on ChO/HRP/CPME was longer than that based on ChO/CPME. 相似文献
17.
Martin Grootveld Christopher J. L. Silwood Edward J. Lynch Ismail Y. Patel David R. Blake 《Free radical research》1999,30(5):351-369
High field proton (1H) NMR spectroscopy has been employed to evaluate the abilities of the antioxidant thiol drug N-acetylcysteine and exogenous cysteine to protect metabolites present in intact inflammatory synovial fluid samples against oxidative damage arising from gamma-radiolysis (5.00 kGy) in the presence of atmospheric O2. Although oxidation of urate to allantoin by radiolytically-generated *OH radical was readily circumventable by pre-treatment of synovial fluids with N-acetylcystine (1.00 or 3.00 × 10-3 mol · dm-3) or cysteine (1.00, 2.00 or 5.00 × 10-3 mol · dm-3), both thiols offered only a limited protective capacity with respect to hyaluronate depolymerisation and the production of formate from carbohydrates in general. Radiolytic products generated from the added thiols (predominantly their corresponding disulphides) were simultaneously detectable in 1H Hahn spin-echo spectra of gamma-irradiated synovial fluids, permitting a quantitative evaluation of the radioprotective capacity of these agents. It is concluded that the multicomponent analytical ability of high field 1H NMR spectroscopy provides much useful molecular information regarding mechanisms associated with the radioprotectant actions of thiols in intact biofluids. 相似文献
18.
Hypochlorous acid (HOCl) is an oxygen-derived species involved in physiological processes related to the defence of the organism that may cause adverse effects when its production is insufficiently controlled. In order to examine its reactivity with potential scavenging molecules from the non steroidal anti-inflammatory drugs (NSAIDs) family, a competition assay based on para-aminobenzoic acid (PABA) chlorination was developed. The original optimised in vitro fluorimetric procedure offered the possibility to determine rate constants (ks) for the reaction with HOCl in physiologically relevant conditions. The specificity of the system was improved by a liquid chromatography (LC) which allows the separation of the drugs and their oxidation products. After determination of the rate constant for PABA chlorination by HOCl (mean±SD in M-1 s-1: 4.3±0.3×103), the applied mathematical model for a chemical competition permits to obtain linear curves from competition studies between several NSAIDs and PABA. Their slopes provided the following rate constants for the different studied drugs: tenoxicam: 4.0±0.7×103, piroxicam: 3.6±0.7×103, lornoxicam: 4.3±0.7×103, meloxicam: 1.7±0.3×104, nimesulide: 2.3±0.6×102. Meloxicam therefore reacted significantly faster than the other oxicams and nimesulide, which is the weakest scavenger of the studied series. The identification of some of the oxidation products by NMR or MS permitted to explore the reaction mechanism and to examine some aspects of the structure/activity relationships for the molecules of the same chemical family. 相似文献
19.
Vascular smooth muscle relaxation by several vasodilators, including acetylcholine (Ach) and ATP, depends on the presence of intact endothelium. Ach is thought to activate muscarinic receptors on endothelium to release an endothelium-derived relaxing factor (EDRF) which brings about relaxation of smooth muscle. In order to assess the role of free radicals in the endothelium-dependent relaxation of blood vessel, we have studied the effect of a spin-trapping agent, phenyl t-butyl nitrone (PBN). on Ach-, ATP-, and sodium nitroprusside-induced relaxation of guinea pig pulmonary artery. Arterial strips were mounted in a 5-ml organ bath containing Krebs solution equilibrated with 95% O2 and 5% CO2 at 37°C. After increasing vascular tone by a synthetic prostaglandin endoperoxide analog (50 ng/ml), the strips relaxed dose-dependently in response to Ach (5 × 10-8M), ATP (1.5 × 10-6M) or sodium nitroprusside (6 × 10-9 M). Removal of the endothelium abolished the relaxation by Ach or ATP, but did not affect the relaxation by sodium nitroprusside. PBN inhibited Ach-induced relaxation of pulmonary artery dose-dependently, but had no effect on relaxations by ATP or sodium nitroprusside. PBN did not block radioligand binding to muscarinic cholinergic membrane receptors on both chick embryonic heart and guinea pig pulmonary artery endothelial cells indicating that it does not block the muscarinic receptors. Spin trapping in combination with electron paramagnetic resonance (EPR) spectral analysis revealed a carbon-centered radical with hyperfine splitting constants of aN = 16.0 G and aβH: = 3.85 G in the lipid extracts of pulmonary artery (0.2-0.4g) incubated with PBN (14mM) and Ach (3 × 10-6M) for 20min. No signal was detected when endothelium was removed. Our data suggest that the endothelium-dependent relaxation of pulmonary artery by Ach is associated with the generation of a free-radical and can be prevented by a spin-trapping agent. ATP, however, relaxes the arterial smooth muscle by a different mechanism. 相似文献
20.
Sulfation of tissue sections which contained nonmetachromatic polysaccharides was done in an equivolume mixture of concentrated sulfuric acid and glacial acetic acid at 0°C. The sections were washed successively in glacial acetic acid and water. Aqueous solutions of toluidine blue, 1.5 × 10-4 M, were used for staining. The procedure produced a metachromatic substrate in sites previously nonmetachromatic. Normal metachromasy was unaffected. There was no loss, damage or distortion of the tissue sections. 相似文献