齿叶白鹃梅叶总黄酮提取物酸水解工艺及水解后苷元含量测定

[目的]确立齿叶白鹃梅叶总黄酮提取物的最佳水解工艺,并对水解后的苷元进行含量测定。[方法]通过正交实验设计确定齿叶白鹃梅叶总黄酮提取物的最佳酸水解条件,利用此酸水解条件,对齿叶白鹃梅叶总黄酮提取物进行酸水解,并采用高效液相色谱法对水解产物中的3种苷元进行含量测定。[结果]确定齿叶白鹃梅叶总黄酮提取物的最佳酸水解条件为:盐酸-甲醇浓度为4.0 mol/L,水解温度为78℃,水解时间为4 h,经过HPLC法测定,水解产物算得到酸法水解后总黄酮苷元的平均含量为58.38%±1.34%,其中木犀草素为12.06%±1.43%,槲皮素为9.66%±0.32%,芹菜素为36.66%±2.34%。[结论]该水解方法可有效水解齿叶白鹃梅叶总黄酮提取物,HPLC测定方法准确、可靠,可作为控制齿叶白鹃梅叶总黄酮水解产物质量的定量依据。     

不同水解方法对葡萄酒厂废渣中齐墩果酸含量的影响

目的：考察不同水解方法对葡萄酒厂废渣中OA含量的影响。方法：采用未水解法、酸解法、酶解法、酶解法＋酸解法4种方法对葡萄榨汁后废渣进行处理，采用醇提法提取出其中的齐墩果酸，并进行含量测定和比较。结果：优选的水解方法为酶解法。结论：所建立的水解方法稳定、可靠，适合于工业生产。     

Receptor reserve of phosphoinositide-coupled muscarinic receptors in mouse hippocampus in vivo

The ability of the partial muscarinic agonist pilocarpine to increase in vivo phosphoinositide (PI) hydrolysis in mouse brain was compared to two full agonists. Pilocarpine increased in vivo phosphoinositide (PI) hydrolysis in cortex, striatum, and to the greatest extent in the hippocampus. Pilocarpine injected either subcutaneously or intracerebroventricularly robustly increased in vivo PI hydrolysis in hippocampus up to 500% of control levels and the increases were blocked by the muscarinic antagonist scopolamine. The increases in vivo PI hydrolysis induced by pilocarpine were 60-75% of the magnitude of the full muscarinic agonists oxotremorine-M and cis-dioxolane. The muscarinic M(1) preferring antagonist pirenzepine potently blocked pilocarpine-induced increases in in vivo PI hydrolysis, consistent with the increase being mediated by M(1) receptors. Since pilocarpine is a relatively weak partial agonist, these data suggest a substantial level of receptor reserve for the PI response in mouse hippocampus.     

Alkaline hydrolysis of methyl, ethyl and n-propyl 4-hydroxybenzoate esters in the liquid and frozen states

The alkaline hydrolysis of the methyl, ethyl and n-propyl esters of 4-hydroxybenzoic acid was studied in the liquid and frozen states in sodium hydroxide solutions. The temperature range was −26 to 60°C. Significant acceleration of the reaction rate was evident in the frozen state compared with rates found at liquid state temperatures. The maximum reaction rate in the frozen state occurred in the temperature range −12 to −10°C. Methyl 4-hydroxybenzoate showed more than a 20-fold increased rate constant from 7.17 × 10⁻⁶ s⁻¹ at 30°C to 1.53 × 10⁻⁴ s⁻¹ at −9°C in a 1.00 × 10⁻² M solution of sodium hydroxide. Rate constants in the liquid and frozen states followed pseudo first-order kinetics over 2–4 half-lives of reaction. Data were fitted to a theoretical model describing the reaction rate in the frozen state as dependent upon the increased concentration of solutes in liquid vesicles in the frozen state and the predicted reduction in the reaction rate constant with temperature decrease. Although the data exhibited similar trends to that predicted by the model, there was frequently a 50% difference in the rates observed compared with those predicted. This study has clearly demonstrated that there is a significantly increased rate of hydrolysis of these esters in the frozen state. This is a further indication that it cannot be assumed that drugs stored in solution will necessarily be stabilized, or their stability enhanced, on freezing. Storage under refrigeration conditions (4–8°C) results in enhanced shelf-lives compared with deep-freeze storage at −20°C under the conditions of this study.     

氨基葡萄糖盐酸盐稳定性及其制备的研究

测定了Ｄ－氨基葡萄糖盐酸盐在酸液中的溶解度及在加热条件下其稳定性，探讨了甲壳素酸水解和甲壳素质量与产品质量、收率之间的关系，对工业化生产Ｄ－氨基葡萄糖盐酸盐的工艺条件优化、质量和收率的提高具有指导意义。     

Cellulose utilization by Clostridium thermocellum: bioenergetics and hydrolysis product assimilation

The bioenergetics of cellulose utilization by Clostridium thermocellum was investigated. Cell yield and maintenance parameters, Y(X/ATP)True = 16.44 g cell/mol ATP and m = 3.27 mmol ATP/g cell per hour, were obtained from cellobiose-grown chemostats, and it was shown that one ATP is required per glucan transported. Experimentally determined values for G(ATP)P-T (ATP from phosphorolytic beta-glucan cleavage minus ATP for substrate transport, mol ATP/mol hexose) from chemostats fed beta-glucans with degree of polymerization (DP) 2-6 agreed well with the predicted value of (n-2)/n [corrected] (n = mean cellodextrin DP assimilated). A mean G(ATP)(P-T) value of 0.52 +/- 0.06 was calculated for cellulose-grown chemostat cultures, corresponding to n = 4.20 +/- 0.46. Determination of intracellular beta-glucan radioactivity resulting from 14C-labeled substrates showed that uptake is different for cellulose and cellobiose (G2). For 14C-cellobiose, radioactivity was greatest for G2; substantially smaller but measurable for G1, G3, and G4; undetectable for G5 and G6; and n was approximately 2. For 14C-cellulose, radioactivity was greatest for G5; lower but substantial for G6, G2, and G1; very low for G3 and G4; and n was approximately 4. These results indicate that: (i) C. thermocellum hydrolyzes cellulose by a different mode of action from the classical mechanism involving solubilization by cellobiohydrolase; (ii) bioenergetic benefits specific to growth on cellulose are realized, resulting from the efficiency of oligosaccharide uptake combined with intracellular phosphorolytic cleavage of beta-glucosidic bonds; and (iii) these benefits exceed the bioenergetic cost of cellulase synthesis, supporting the feasibility of anaerobic biotechnological processing of cellulosic biomass without added saccharolytic enzymes.     

Phospholipase C-gamma1 in tumor progression

The vast majority of cancer morbidity and mortality arises from tumor progression beyond the primary tumor site. Unfortunately, most therapies are not effective for advanced stage disease with regional extension or distant metastases. Thus, new treatments are needed to target rate limiting steps in tumor progression. The ability of cancers to invade and metastasize requires the acquisition of specific cell behaviors that enable the cell to escape from the localized site, breach the defined boundaries, reach a hospitable ectopic site and grow in this new locale. Recently, dysregulation of cell motility as stimulated by various extracellular factors has gained credence as a rate-limiting alteration in tumor progression in carcinomas and some other solid tumors. This has focused attention on initiators of signaling cascades that regulate tumor migration. In this effort, one molecule, phospholipase C-γ1 (PLCγ), has been shown to function as a key molecular switch. This revised version was published online in July 2006 with corrections to the Cover Date.     

Tetanus toxin modulates serotonin transport in rat-brain neuronal cultures

As has been previously described, tetanus toxin (TeTx) and its H_C fragment inhibit the sodium-dependent 5-hydroxytryptamine (5-HT) uptake in rat-brain synaptosomes, probably through a kinase mechanism affecting the 5-HT transporter. Now, the inhibition of 5-HT uptake in neurons in primary culture by TeTx in a dose-dependent manner is described in this work. This effect is also produced by the nontoxic C-terminal fragment of the TeTx heavy chain (Hc-fragment), indicating that 5-HT uptake inhibition is a consequence of the toxin binding to the plasmatic membrane and not to its catalytic activity. This conclusion is supported by the fact that the 5-HT accumulation was not inhibited by the light chain of TeTx or the toxoid, and was even potentiated by botulinum neurotoxin A. These results correlate with the activation of phosphoinositide-phospholipase C activity in the cultures used in this study, this activity only being enhanced by TeTx and by its Hc-fragment. On the other hand, the use of tyrosine phosphorylation modulators indicates that both Na₃VO₄ and basic fibroblast growth factor (bFGF) produce an enhancement of 5-HT uptake in this system, which is also sensitive to TeTx inhibition. On the other hand, genistein alone is able to reduce the 5-HT transport in cultured neurons, and this effect did not appear to be additive to that elicited by TeTx. This result suggests that TeTx and genistein may share some events in their respective mechanisms of action. Furthermore, the incubation at different concentrations of 12-0-tetradecanoylphorbol 13-acetate (TPA) confirms the involvement of protein kinase C (PKC) in 5-HT transport modulation in rat-brain neuronal primary cultures. In summary, we shall demonstrate in this work that TeTx induces, through its Hc fragment, an inhibition of both basal and stimulated serotonin uptakes in primary neuronal cultures, in parallel to the activation of phosphoinositide-phospholipase C activity and PKC activation.     

Acyloxymethyl acidic drug derivatives: in vitro hydrolytic reactivity

A series of acyloxymethyl drug derivatives of the NH-acidic drugs, phenytoin and theophylline and of the carboxylic acid drugs, thioctic acid and indomethacin, were prepared in order to determine the effect of varying the nature of the drug on the in vitro rate of hydrolysis catalyzed by porcine liver esterase and human plasma. The acyl portion was comprised of either valeric acid (val) or γ-linolenic acid (GLA). With the exception of some GLA prodrugs, the derivatives displayed first-order kinetics in both enzyme systems. The NH-acidic drug derivatives were hydrolyzed faster than the carboxylic drug derivatives by porcine liver esterase and human plasma. It was found that the short chain valeric acid derivatives were hydrolyzed faster than the GLA derivatives. The rates of hydrolysis for the relatively smaller prodrugs of theophylline and thioctic acid were greater than the rates of hydrolysis for the bulkier phenytoin and indomethacin prodrugs indicating steric hindrance was important. The lipophilicity index, log K, of the valeric acid drug derivatives was plotted against the logarithm of the hydrolysis rate constant, k, and it was observed that log k decreased with an increase in log K. A comparison of these results with those of previous studies where the alkyl and acyl moieties were varied of acyloxyalkyl theophylline derivatives has provided a rationale, based on lipophilicity, for the structure of a prodrug to be designed based on an in vitro desired rate of hydrolysis.