Reaction mechanism of isomaltooligosaccharides synthesis by α-glucosidase fromAspergillus carbonarious

Summary An -glucosidase fromAspergillus carbonarious CCRC 30414 was employed for investigating the enzymatic synthesis of isomaltooligosaccharides from maltose. The enzyme transferred a glucose unit from the nonreducing end of maltose and other -linked glucosyl oligosaccharides to glucose and other glucosyl oligosaccharides which function as accepting co-substrates. The transfer of a glucose unit occurs most frequently to the 6-OH position of the nonreducing end of acceptor, but transfer to 4-OH position also occurs. Treatment of 30 % (w/v) maltose with the enzyme under optimum conditions afforded more than 50% isomaltooligosaccharides.     

DNA binding site specificity of the Neurospora global nitrogen regulatory protein NIT2: Analysis with mutated binding sites

NIT2, a positive-acting regulatory protein in Neurospora crassa, activates the expression of a series of unlinked structural genes that encode nitrogen catabolic enzymes. NIT2 binding sites in the promoter regions of nit3, alc and lao have at least two GATA sequence elements. We have examined the binding affinity of the NIT2 protein for the yeast DAL5 wild-type upstream activation sequence UAS_NTR, which contains two GATA elements, and for a series of mutated binding sites, each differing from the wild-type site by a single base. Substitution for individual nucleotides within 5′ or 3′ sequences that flank the GATA elements had only modest effects upon NIT2 binding. In contrast, nearly all substitutions within the GATA elements almost completely eliminated NIT2 binding, demonstrating the importance of the GATA sequence for NIT2 binding. Four high-affinity binding sites for the NIT2 protein were found within a central region of the nit-2 gene itself.     

Induction of apoptosis and c-myc in L1210 lymphocytic leukemia cells by adenosine

High concentrations of adenosine (Ado), when added to L1210 lymphocytic leukemia cells, resulted in apoptosis or programmed cell death. The apoptotic process was accompanied by distinct morphological changes including chromatin condensation and blebbing of plasma membranes. Extensive DNA fragmentation was correlated with Ado concentrations. Furthermore, apoptosis in these cells was preceded by an early but transient expression of c-myc proto-oncogene, and was not influenced by homocysteine thiolactone added to the cells. Since severe combined immunodeficiency (SCID) is associated with a deficiency of adenosine deaminase, leading to defects in both cellular and humoral immunity, Ado-induced apoptosis may thus be a contributing factor in the pathology of SCID.     

Identification and characterization of glucocorticoid receptors in liver of nude mice

Glucocorticoids regulate the expression of many liver-specific genes via glucocorticoid receptors. The presence of glucocorticoid receptors in liver has been reported in many mammalian species but not in nude mice. In the present study, we demonstrate the presence of specific glucocorticoid receptors in nude mouse liver. The binding of ligands to these receptors could be completely inhibited by RU486, and partially blocked by hydrocortisone and progesterone, whereas estrogen and testosterone had no effect. Hydrocortisone down-regulated the level of glucocorticoid receptors in livers of nude mice and correspondingly enhanced the activities of tyrosine aminotransferase and -glutamyltransferase. Our results indicate that glucocorticoid receptors in nude mouse liver are specific, fully functional, and present at levels 28.5-fold higher than in the liver of normal inbred mice. We suggest that the nude mouse is a valuable model for studies of hepatic glucocorticoid action and may provide a clue to a putative hepatic-thymic interaction.     

Action of lipolytical enzymes in biphasic organic-aqueous systems: dynamics of the irreversible Michaelis-Menten reaction

Through simple model analysis, the mass action kinetic model for lipolytic enzymes in biphasic aqueous-organic systems can be simplified using the quasi-steady state assumption (or the quasi-equilibrium state assumption) for the adsorbed enzyme E* or the enzyme-substrate complex E*S. Some parameter combinations leading to the above assumptions are derived confirmed by full numerical integration of the whole enzymatic process. The results may be classified into three categories: (1) the quasi-equilibrium state assumption for E*, (2) the quasi-steady state assumption for E*, and (3) the quasi-steady state assumption for E*S. Further simplification for both E* and E*S is also discussed. (c) 1993 Wiley & Sons, Inc.     

细菌Ⅵ型分泌系统效应蛋白的装配及功能的研究进展

蛋白分泌作为细胞之间传递信号的途径之一,在微生物生存竞争中也扮演着重要的角色。革兰氏阴性菌可以通过Ⅵ型分泌系统(type Ⅵ secretion system, T6SS)将效应蛋白传递至胞外或原核和真核微生物中,从而介导微生物间的竞争或宿主-细菌的相互作用,最终建立竞争优势。本文主要总结了T6SS的结构与组成,并重点对效应蛋白的装配以及其与免疫蛋白的作用机制的研究进展进行阐述,为以后靶向T6SS抗菌药物的研制提供新思路。     

放牧对祁连山高寒典型草原土壤质量的影响

土壤质量是维持陆地生态系统稳定性与功能多样性的基础。放牧作为草地资源最广泛的利用方式之一,其对草地土壤质量的影响却缺乏量化标准,且两者之间的作用机理尚不明确。以祁连山高寒草原两个季节性牧场为研究对象,结合生态系统耦合与生态系统多功能性,探究了放牧对高寒草原土壤质量的影响与潜在机制。试验结果表明：基于最小数据集,不同放牧率下土壤质量指数差异显著(P<0.05),冬季牧场和春秋季牧场放牧率分别在2.45头月^-1 hm^-2和0.80头月^-1 hm^-2时土壤质量指数最高。土壤速效磷、有机碳、氮磷比和土壤pH是决定冬季牧场土壤质量的关键因子,而春秋季牧场中则是土壤有机碳、碳氮比和土壤pH;两个季节性牧场土壤质量指数与物种丰富度指数(P<0.05)和香浓维纳多样性指数(P<0.0001)呈显著正相关。高寒草原季节性牧场放牧地植物群落物种多样性与土壤因子耦合度在0.67—0.81之间,平均耦合度为0.74,属于中度协调;随着放牧率的增加,生态系统多功能性指数逐渐减低且与土壤质量指数变化趋势相似,...     

海洋生物礁类型、生态功能及其生态修复

海洋生物礁是由具有造礁能力的海洋生物聚集而成的一种三维礁体结构,其形成改变了海底地貌、增加了不同尺度上的地形复杂性,为其他海洋生物提供了栖息地并维持了生物多样性。近年来,由于自然因素和人为因素影响,海洋生物礁受到了严重威胁,已成为海洋生态保护和修复领域的重要研究对象。综述了海洋生物礁的类型、生态功能及其生态修复的研究进展。根据形成海洋生物礁的优势造礁生物种类,将海洋生物礁分为海藻礁、海绵礁、刺胞动物礁、贝类礁和多毛类礁,其优势造礁生物分别是珊瑚藻和仙掌藻、钙质海绵和硅质海绵、造礁珊瑚、牡蛎、龙介虫。目前国内对海洋生物礁的全面了解相对较少,主要集中在珊瑚礁和牡蛎礁。海洋生物礁的生态功能主要有海岸防护、提供栖息地、净化水体、固碳作用和能量耦合等。全球变暖和海洋酸化等全球气候变化以及海洋污染、破坏性渔业捕捞、海岸工程、水产养殖和敌害生物等自然和人为因素对海洋生物礁构成了严重威胁。海洋生物礁的生态修复方法分为两类：在退化生物礁区投放造礁生物逐渐成礁,投放人工礁体补充造礁生物逐渐成礁。针对海洋生物礁保护和修复的需要,提出下一步应加强海洋造礁生物生态特征、海洋造礁生物种群丧失因素和海洋生物礁保护与...     

Determination of nalbuphine by high-performance liquid chromatography with ultraviolet detection: application to human and rabbit pharmacokinetic studies

A rapid, sensitive, precise and accurate high-performance liquid chromatographic assay with ultraviolet detection was developed for the determination of nalbuphine in human, rabbit, pig and dog plasma. It is comprised of only a one-step extraction procedure with hexane-isoamyl alcohol at pH 9.25 and reversed-phase chromatography on a μPorasil column. The recoveries of nalbuphine and ethylmorphine (internal standard) were greater than 86%. Calibration graphs were linear over the concentration range 0.75–150 ng/ml with a coefficient of variation, both within-day and between-day, of less than 10% at any level. The limit of quantitation was 0.75 ng/ml of plasma based on a signal-to-noise ratio of 3. Seven other clinically used analgesics were investigated to check for potential interferences and their analytical conditions. The specificity of this assay was checked with a metabolite of nalbuphine (noroxymorphine). Nalbuphine in plasma did not decompose significantly at −20°C for six weeks. Pharmacokinetic application in three surgical patients and four rabbits revealed that nalbuphine followed a linear three-compartment model with two distribution phases. The two distribution and one elimination half-lives and the plasma clearance of nalbuphine were 0.9, 5.8 and 157 min and 370 ml/min in human, and 3.5, 28 and 117 min and 21 166 ml/min in rabbits.