共查询到20条相似文献,搜索用时 31 毫秒
1.
Jinsheng Xu John H. Lamb Rebecah Jukes Wendy M. Purcell David E. Ray 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1999,732(2):307
This study presents a HPLC method for the separation and purification of p-bromophenylacetylurea (BPAU) and its metabolites. The method effectively separated and purified BPAU and its metabolites. Three metabolites of BPAU, M1, M2 and M3 were characterised by mass spectroscopy and nuclear magnetic resonance. They are named as N′-hydroxy-p-bromophenylacetylurea, 4-(4-bromophenyl)-3-oxapyrrolidine-2,5-dione and N′-methyl-p-bromophenylacetylurea, respectively. The major metabolic pathways of BPAU were proposed. The establishment of the HPLC method and characterisation of BPAU metabolites make it possible for further pharmacokinetic studies to explore the mechanism of BPAU-induced delayed neuropathy. 相似文献
2.
Olivier Tranquet Luc Saulnier Jean-Pierre Utille John Ralph Fabienne Guillon 《Phytochemistry》2009,70(11-12):1366-1373
p-Coumaric acid is one of the predominant phenolic acids acylating the cell walls of grasses; p-coumarates are mainly esterified by lignins and arabinoxylans. Here we describe the production and characterisation of two monoclonal antibodies against p-coumarates.The 5-O-pCou-Ara(1 → 4)Xyl was chemically synthesized and conjugated to a carrier protein. Two interesting antibodies were obtained, hereinafter named INRA-COU1 and INRA-COU2. The specificity of these monoclonal antibodies has been evaluated using competitive-inhibition assays with different oligosaccharides and phenolic compounds. INRA-COU1, recognized free p-coumaric acid or p-coumarate esters. INRA-COU1 did not react with any of the other hydroxycinnamic acids and related compounds found in plants. INRA-COU2, only recognizes esterified p-coumarate. These antibodies were used to study the localization of p-coumarates in the cell walls of grasses. Immunocytochemical analyses indicated noticeable amounts of p-coumarate in the cell walls of the aleurone layer of wheat grain, in the epiderm of cereal straw, and in the exoderm of wheat root.The use of these antibodies will contribute to a better understanding of the organisation and developmental dynamics of cell walls in Graminaceae. 相似文献
3.
Crystal structures of cyclomaltohexaose (α-cyclodextrin) complexes with p-chlorophenol and p-cresol have been determined by single-crystal X-ray diffraction studies. The space group of the α-cyclodextrin–p-chlorophenol complex is P212121 with unit cell dimensions of a=15.299(3), b=24.795(5), c=13.447(5) Å, and that of the α-cyclodextrin–p-cresol complex is P21 with unit cell dimensions of a=7.927(7), b=13.568(7), c=24.54(1) Å, β=90.41(8)°. In spite of the similar structures of guest molecules, both complexes have different inclusion modes and packing structures. 相似文献
4.
The direction in which both sodium p-nitrophenolate and p-nitrophenol penetrate the cyclohexaamylose cavity in aqueous solution has been examined by 13C-nmr. Both sodium p-nitrophenolate and p-nitrophenol penetrate the cavity asymetrically and quite specifically nitro group first with the phenolic oxanion or hydroxyl group pointing out into solution as evidenced by the nature of the changes in the meta-carbon-13 shifts. The stoichiometries of the complexes can be defined for various host-to-guest molar ratios. Finally, the potential of these cycloamylose complexes as models for studying the effects of intermolecular interaction of the enzyme substrate type on the 13C-nmr of both host and guest molecules is discussed. 相似文献
5.
Keith A. Brown Colin Ratledge 《Biochimica et Biophysica Acta (BBA)/General Subjects》1975,385(2):207-220
p-Aminosalicylic acid inhibits growth of Mycobacterium bovis BCG and Mycobacterium smegmatis more effectively if cells are growing with a sufficiency of iron (> 1 μg Fe/ml) in the medium than if cells are deficient in iron (<0.1 μg Fe/ml). In iron-deficient cultures formation of mycobactin, an ionophore for iron transport, is strongly inhibited by p-aminosalicylic acid. Uptake of iron into cell suspensions is also inhibited and the activity of several iron-containing enzymes declines in cells exposed to p-aminosalicylic acid during their growth. p-Aminosalicylic acid is about 50 times more effective towards a mutant of M. smegmatis which required mycobactin under iron-deficient growth conditions than towards the wild-type parent. p-Aminosalicylate is taken up into cells by an active process independent of the salicylate uptake system, possibly by the route used for assimilation of p-aminobenzoate. (This could account for why p-aminobenzoic acid, but not salicylic acid, antagonizes the action of p-aminosalicylic acid.) With iron-deficient cells, salicylate assimilation is about 50 times greater than either p-aminosalicylate or p-aminobenzoate but with iron-sufficient cells and with the mycobactin mutant salicylate uptake is negligible whereas p-aminobenzoate and p-aminosalicylate uptakes are unaffected. p-Aminosalicylic acid at 3.3 mM (500 μg/ml) partially inhibits the uptake of both p-aminobenzoate and, if it is occuring, that of salicylate as well. As p-aminosalicylic acid is always more effective when the intracellular concentration of salicylic acid is low, it probably acts as an anti-metabolite of salicylic acid, not, however, by inhibiting the conversion of salicylic acid to mycobactic, but probably somewhere along the metabolic pathway of iron uptake. 相似文献
6.
Ganga-Ramu Vasanthakumar Basanagoud S. Patil Vommina Venkata Suresh Babu 《Letters in Peptide Science》2002,9(4-5):207-209
A simple method for the synthesis of several amino acidbenzyl ester p-toluenesulfonate salts from thecorresponding amino acid and benzyl alcohol in presence of p-toluenesulfonic acid accelerated with microwave irradiation isdescribed. Under similar condition, the amino acid benzyl esterhydrochloride salts have also been obtained by using thionylchloride instead of p-toluenesulfonic acid in good yieldand purity. 相似文献
7.
A silver ion-loaded microparticulate cation-exchange resin column has been used for high-performance liquid chromatographic (HPLC) separation of the p-nitrophenacyl esters of several series of closely related prostaglandins: 8-iso-PGE2, 11-epi-PGE2, 5-trans-PGE2, PGE2, PGF1α, PGE1, and PGF2α, PGA2 and PGB2; 15 (R)-methyl-PGE2 and 15 (S)-methyl-PGE2; 5-trans-PGA2 and PGA2; and 5-trans-PGF2α and PGF2α. The properties of this column are compared with those of silica-gel and reversed-phase columns. 相似文献
8.
L. A. Decosterd A. Karagiannis J. -M. Roulet N. Blaz M. Appenzeller T. Buclin P. Vogel J. Biollaz 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1997,703(1-2)
PAH (N-(4-aminobenzoyl)glycin) clearance measurements have been used for 50 years in clinical research for the determination of renal plasma flow. The quantitation of PAH in plasma or urine is generally performed by colorimetric method after diazotation reaction but the measurements must be corrected for the unspecific residual response observed in blank plasma. We have developed a HPLC method to specifically determine PAH and its metabolite NAc-PAH using a gradient elution ion-pair reversed-phase chromatography with UV detection at 273 and 265 nm, respectively. The separations were performed at room temperature on a ChromCart® (125 mm×4 mm I.D.) Nucleosil 100-5 μm C18 AB cartridge column, using a gradient elution of MeOH–buffer pH 3.9 1:99→15:85 over 15 min. The pH 3.9 buffered aqueous solution consisted in a mixture of 375 ml sodium citrate–citric acid solution (21.01 g citric acid and 8.0 g NaOH per liter), added up with 2.7 ml H3PO4 85%, 1.0 g of sodium heptanesulfonate and completed ad 1000 ml with ultrapure water. The N-acetyltransferase activity does not seem to notably affect PAH clearances, although NAc-PAH represents 10.2±2.7% of PAH excreted unchanged in 12 healthy subjects. The performance of the HPLC and the colorimetric method have been compared using urine and plasma samples collected from healthy volunteers. Good correlations (r=0.94 and 0.97, for plasma and urine, respectively) are found between the results obtained with both techniques. However, the colorimetric method gives higher concentrations of PAH in urine and lower concentrations in plasma than those determined by HPLC. Hence, both renal (ClR) and systemic (ClS) clearances are systematically higher (35.1 and 17.8%, respectively) with the colorimetric method. The fraction of PAH excreted by the kidney ClR/ClS calculated from HPLC data (n=143) is, as expected, always <1 (mean=0.73±0.11), whereas the colorimetric method gives a mean extraction ratio of 0.87±0.13 implying some unphysiological values (>1). In conclusion, HPLC not only enables the simultaneous quantitation of PAH and NAc-PAH, but may also provide more accurate and precise PAH clearance measurements. 相似文献
9.
西洋参(Panax quinquefolium L.)栽培中存在严重的连作障碍现象,前期发现p-香豆酸在以滤纸片为基质的条件下,能够显著抑制西洋参胚根的生长。为了明确p-香豆酸在土壤基质中对种胚的化感活性以及对成株西洋参生长的作用及生理机制,以自然土壤为基质,观察p-香豆酸作用后种胚的生长情况;采用室内水培试验,观察p-香豆酸作用下2年生西洋参种根从出苗至结果期的生长及部分生理指标的变化。种胚生长实验在土壤中分别添加0.0024、0.012、0.06、0.3、1.5、7.5 mg/g的p-香豆酸,处理7 d后测定西洋参种胚的胚根长和胚芽长。水培试验中全营养液中分别添加0.012 mg/mL、0.06 mg/mL、0.3 mg/mL p-香豆酸,处理后每隔5 d测定植株叶片展开情况、株高、冠幅等生长指标;于展叶期(10 d)、现蕾期(20 d)、结果期(30 d)测定地上部分及新生须根的生物量,同时测定新生须根苯丙氨酸解氨酶(PAL)活力;叶片完全展开后测定植株净光合速率(Pn)、表观电子传递速率(ETR)和最大光化学效率(Fv/Fm)等光合特性参数。结果表明,土壤中添加0.0024-7.5 mg/g p-香豆酸西洋参胚根长度降低28.52%-100%,胚芽长度降低1.09%-100%,并呈现一定的剂量抑制效应。实验浓度内的p-香豆酸可显著抑制西洋参植株地上部分生长,推迟展叶期;结果期地上部生物量比对照降低17.17%-54.55%(P < 0.05,Dunnett-t test);同时,叶片的Pn、ETR受到抑制(P < 0.05),但Fv/Fm不变;对须根的影响主要表现为0.06 mg/mL p-香豆酸处理组在展叶期PAL酶活力提高69.05%,之后PAL活力和生物量均比对照下降,浓度增加至0.3 mg/mL时整个培养期内PAL酶活力和生物量均低于对照。由此推论,根系环境中的p-香豆酸在自然土壤中对西洋参种胚具有显著抑制其生长的化感作用;对成株西洋参的作用主要为抑制地上部分生长,并通过降低成株西洋参叶片光合能力,从而表现出明显的化感作用,0.06 mg/mL p-香豆酸诱导须根PAL酶活力先升高再降低并最终降低生物量的结果也表明p-香豆酸是西洋参根系生长的胁迫因素。结果证实p-香豆酸对西洋参种胚和成株的生长均具有自毒作用,其抑制生长的生理机制在于抑制叶片的光合作用。 相似文献
10.
Jia-Shun GONG Wei-Peng MA Jun-Xue PU Shu-Guan XU Shuang-Qing ZHENG Chun-Jie XIAO 《生物工程学报》2006,22(5):800-805
The conversion of exogenous p-hydroxybenzaldehyde into p-hydroxy-methyl-phenol-β-D-glucoside (gastrodin) was studied using cell suspension cultures of Datura tatula L. The chemical structure of the synthesized gastrodin was identified on the basis of spectral analysis and chemical evidence. The procedure of conversion of p-hydroxybenzaldehyde into gastrodin by D. tatula L. cell suspension cultures was established. The synthesized gastrodin (II) was isolated from the ferment liquor and identified by spectral analysis. Simultaneously, the p-hydroxybenzyl alcohol (I) that was converted through biotransformation of p-hydroxybenzaldehyde by cell suspension cultures of D. tatula L. was also isolated and identified. The efficiency of glucosylation of p-hydroxybenzaldehyde was remarkably enhanced by the addition of salicylic acid (0.1 mg/L) and the maintenance of low pressure (0.001 MPa) in a 25-L airlift loop bioreactor. The biotransformation of exogenous p-hydroxybenzaldehyde to gastrodin using cell suspension cultures of D. tatula L. is a promising approach. 相似文献
11.
Thomas F. Holzman Christine C. Chung Rohinton Edalji David A. Egan Margaret Martin Earl J. Gubbins Grant A. Krafft Gary T. Wang A. Mitchel Thomas Saul H. Rosenberg et al. 《Journal of Protein Chemistry》1991,10(5):553-563
The kinetic behavior andpH-stability of recombinant human renin was analyzed using a new fluorogenic substrate based on the normal P6-P3 renin cleavage sequence in human angiotensinogen. The design of this fluorogenic substrate makes possible, for the first time, direct monitoring of the kinetics of proteolytic conversion of prorenin to renin. ThepH-stability profile for renin, measured with the substrate at 25°C, indicated a broad plateau of stability betweenpH 6.0 and 10.0. Analysis of thepH-activity profile of renin for the substrate indicated a minimumK
m
(1.8 µM) atpH 7.4 and a maximumV
m
betweenpH 7.4 and 8.0. The thermodynamics of the binding of a novel, soluble, peptidomimetic inhibitor to renin indicated it is possible to retain the tight-binding characteristics and enthalpy contributions to binding of larger peptide-derived inhibitors, while reducing inhibitor size and entropic contributions to binding. A novel derivative of the fluorogenic substrate, containing a 3-methyl histidine substitution at the P2 site, was used to test the recent hypothesis that renin functions by virtue of substrate-directed catalysis. 相似文献
12.
p14ARF对人黑色素瘤细胞增殖的影响及其作用机理的初探 总被引:2,自引:0,他引:2
ARF(alternative reading frame)作为INK4a/ARF的β转录产物,能够稳定p53, 诱导细胞周期阻断或凋亡.利用高表达p14ARF的人黑色素瘤细胞模型,探讨了ARF抑制细胞增殖的分子作用机理.研究发现p14ARF高表达能将细胞周期阻断在G1和G2期, p53, p21cip1和p27kip1蛋白水平明显增强, 而p-ERK1/2,CyclinD1和CyclinE蛋白水平下降, 明显抑制细胞生长. 提示p14ARF能通过ERK(extracellular signal-regulated kinase)信号通路相互协调作用抑制A375细胞增殖. 相似文献
13.
Effects of pH and ligand binding upon the conformation of Cellobiohydrolase I (CBHI) and endoglucanase I (EGI) from Trichoderma pseudokiningii S-38 have been studied by circular dichroism measurements. In the high-pH range (6–9), increasing pH resulted in a similar conformational change occurring in free CBHI and EGI, while such treatment gave different changes of the two enzyme conformations in the presence of cellobiose. On the other hand, in the low-pH region, with both CBHI an EGI in the active form, decreasing pH resulted in a large conformational change of free EGI compared to that of free CBHI, whereas ligand binding resulted in a similar change of both CBHI and EGI, independent of pH change. 相似文献
14.
Xinghui Qiu Qiuzan Zhong Mei Li Wenqin Bai Baotong Li 《International biodeterioration & biodegradation》2007,59(4):297-301
Ochrobactrum sp. B2, a methyl parathion-degrading bacterium, was proved to be capable of using p-nitrophenol (PNP) as carbon and energy source. The effect of factors, such as temperature, pH value, and nutrition, on the growth of Ochrobactrum sp. B2 and its ability to degrade p-nitrophenol (PNP) at a higher concentration (100 mg l−1) was investigated in this study.The greatest growth of B2 was observed at a temperature of 30 °C and alkaline pH (pH 9–10). pH condition was proved to be a crucial factor affecting PNP degradation. Enhanced growth of B2 or PNP degradation was consistent with the increase of pH in the minimal medium, and acidic pH (6.0) did not support PNP degradation. Addition of glucose (0.05%, 0.1%) decreased the rate of PNP degradation even if increased cell growth occurred. Addition of supplemental inorganic nitrogen (ammonium chloride or ammonium sulphate) inhibited PNP degradation, whereas organic nitrogen (peptone, yeast extract, urea) accelerated degradation. 相似文献
15.
16.
The ability of grape skins to catalyze in vitro conversion of p-coumaric acid to the more potent antioxidant caffeic acid was studied. Addition of different concentrations of p-coumaric to red grape skins (Cabernet Sauvignon) resulted in formation of caffeic acid. This caffeic acid formation (Y) correlated
positively and linearly to p-coumaric acid consumption (X): Y = 0.5 X + 9.5; R
2 = 0.96, P < 0.0001. The kinetics of caffeic acid formation with time in response to initial p-coumaric acid levels and at different grape skin concentrations, indicated that the grape skins harboured an o-hydroxylation activity, proposedly a monophenol- or a flavonoid 3′-monooxygenase activity (EC 1.14.18.1 or EC 1.14.13.21).
The K
m of this crude o-hydroxylation activity in the red grape skin was 0.5 mM with p-coumaric acid. 相似文献
17.
R.J.E Grouls E.W Ackerman H.H.M Korsten L.J Hellebrekers D.D Breimer 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1997,694(2):1313
For the determination of the logarithmic partition coefficients between n-octanol and water (log Po/w) of local anesthetics, the pH of the aqueous phase needs to be adjusted to high values to ensure that the local anesthetics are in the unionized form. Using the shake-flask or the stir-flask method, this high pH may catalyze hydrolysis, leading to increasing amounts of impurities in time. These impurities exclude non-selective quantification methods like UV spectrometry and require repetitive quantitative analysis of both liquid phases resulting in a tedious and time-consuming method. A rapid reversed-phase HPLC method was developed to measure log Po/w of the local anesthetics N-butyl-p-aminobenzoate, methyl-p-aminobenzoate, benzocaine, procaine, mepivacaine, prilocaine, lidocaine, bupivacaine, etidocaine, tetracaine and oxubuprocaine. 相似文献
18.
Alexander A. Kortt John E. Burns J. Bruce Caldwell Teresa Ferro Phillip M. Strike 《Journal of Protein Chemistry》1991,10(2):183-188
The primary structure of acidic trypsin inhibitor-2a (WBTI-2a,pI 5.9) fromPsophocarpus tetragonolobus (L.) DC seed was determined. This inhibitor consists of a single polypeptide chain of 180 amino acids including four half-cystine residues and has an N-terminal residue of pyroglutamic acid. The sequence of WBTI-2a,pI 5.9, showed 84% identity to acidic trypsin inhibitor-2 (WBTI-2,pI 5.1) but only 57% identity to the basic trypsin inhibitor (WBTI-1,pI 8.9) and 50% identity to the chymotrypsin inhibitor of winged bean. The data indicate that winged bean seed contains a family of three Kunitz-type inhibitors which have about 50% identity. 相似文献
19.
The effects of adding D2SO4, and p-toluenesulfonic acid-d to D-cellobiose dissolved in D2O were investigated at 23 °C by plotting 13C NMR chemical shift changes (Δδ) against the acid to D-cellobiose molar ratio. 13C Chemical shifts of all 18 carbon signals from α and β anomers of D-cellobiose showed gradual decreases due to increasing acidity in aqueous D2SO4 medium. The C-1 of the α anomer showed a slightly higher response to increasing D+ concentration in the surrounding. In the aqueous p-toluenesulfonic acid-d medium, C-6′ and C-4′ carbons of both α, and β anomeric forms of D-cellobiose are significantly affected by increasing the sulfonic acid concentrations, and this may be due to a 1:1 interaction of p-toluenesulfonic acid-d with the C-6′, C-4′ region of the cellobiose molecule. 相似文献