An improved assay for UDPglucose pyrophosphorylase and other enzymes that have nucleotide products

UDPglucose pyrophosphorylase catalyses the interconversion UDPglucose plus pyrophosphate and glucose 1-phosphate plus UTP. Several assay methods for this enzyme have been described but the only one that can be used to investigate the specificity with respect to various UDPsugars is based on coupling to UTP formation. This assay employs phosphoglycerate kinase to catalyse the formation 1,3-bisphosphoglycerate which is then used to oxidise NADH in the presence of glyceraldehyde 3-phosphate dehydrogenase. We have found that the activity of phosphoglycerate kinase towards UTP is low which limits the usefulness of the assay to very low rates, in agreement with the published recommendation of Hansen et al.⁵. Here it is shown that the dynamic range of the assay is increased by more than five fold on addition of nucleoside diphosphate kinase and ADP, which convert UTP to the preferred phosphoglycerate kinase substrate, ATP. It is also shown that the improved assay is suitable for enzymes with other nucleotide triphosphate products.     

Developmental actions of natriuretic peptides in the brain and skeleton

The concept that atrial natriuretic peptide (ANP) and the closely related peptides BNP and CNP might be involved in the ontogeny of several organ systems emerged in the late 1980s. While many of the reported in vitro actions have not been examined in the context of organ development in vivo, recent studies demonstrate that mice which lack or overexpress natriuretic peptides or receptors exhibit pronounced skeletal growth defects. This article discusses how natriuretic peptides and other factors appear to regulate bone growth as an example of how natriuretic peptides might participate in the ontogeny of other organ systems. Evidence indicating that natriuretic peptides regulate neural development is then reviewed. Natriuretic peptides and receptors exhibit complex expression patterns in the developing nervous system, where they have been shown to act on neural cells as early as at the embryonic neural tube stage. Interestingly, both bone and brain growth appear to utilize primarily CNP and the CNP-specific type B receptor, and perhaps the type C receptor. In vitro data indicate that CNP may act on developing neurons, astrocytes and Schwann cells like a classical growth factor, regulating proliferation, patterning, phenotypic specification, survival and axonal pathfinding. Natriuretic peptides might also have roles in the vascularization of the embryonic brain, establishment of the blood-brain and blood-nerve barriers, and perhaps in nerve regeneration.Received 13 April 2004; received after revision 20 May 2004; accepted 27 May 2004     

Directed evolution of enzymes for biocatalysis and the life sciences

Engineering the specificity and properties of enzymes and proteins within rapid time frames has become feasible with the advent of directed evolution. In the absence of detailed structural and mechanistic information, new functions can be engineered by introducing and recombining mutations, followed by subsequent testing of each variant for the desired new function. A range of methods are available for mutagenesis, and these can be used to introduce mutations at single sites, targeted regions within a gene or randomly throughout the entire gene. In addition, a number of different methods are available to allow recombination of point mutations or blocks of sequence space with little or no homology. Currently, enzyme engineers are still learning which combinations of selection methods and techniques for mutagenesis and DNA recombination are most efficient. Moreover, deciding where to introduce mutations or where to allow recombination is actively being investigated by combining experimental and computational methods. These techniques are already being successfully used for the creation of novel proteins for biocatalysis and the life sciences.Received 8 June 2004; received after revision 22 July 2004; accepted 2 August 2004     

我国稀土发光材料科学技术发展回顾与展望

本文对我国稀土发光材料科学和技术的发展，从机遇和起步，跟踪和引进，自主发展，带动相关科技和产业化发展进行较全面回顾，稀土发光材料已经成为我国信息显示，绿色照明工程，医疗健康，光电子等领域的支柱材料，对比与发达国家的差距和存在问题，对进入新世纪和WTO后，所面临的新机遇-新挑战，以及蓬勃发展前景予以展望。     

The effect of ethanol on the biosynthesis and regulation of opioid peptides

Summary Alcoholism and alcohol abuse are serious health problems. Alcohol is known to influence the activity of a number of biological systems, for example the hormonal and neuronal systems. One of the biological systems whose activity is greatly influenced by alcohol is the endogenous opiate system. Alcohol modifies the function of both opiate receptors and opioid peptides. In fact it has been proposed that many of the effects of ethanol are mediated by its effects on the endogenous opiate system. This review will present results from various laboratories on the effects of acute and chronic ethanol treatments on various species, and on the release, biosynthesis and post-translational processing of the endorphins, enkephalins and dynorphins, the three known families of endogenous opioid peptides. Furthermore, the effect of acute and chronic ethanol consumption on the -endorphin system in man, and the possible implications of the functional activity of the endogenous opiate system for the genetic predisposition to alcoholism will be discussed.     

The effect of corticotropin-releasing factor and pro-opiomelanocortin-derived peptides on the phagocytosis of molluscan hemocytes

The effect of corticotropin-releasing factor (CRF) and pro-opiomelanocortin (POMC)-derived peptides on hemocyte phagocytosis in two molluscs,Planorbarius corneus andViviparus ater was studied. The peptides and related fragments examined are those which have been shown to influence hemocyte motility in the two species. The results obtained revealed that the effects on phagocytosis are not directly correlated with previous findings on cell motility. Furthermore, the mode of action of an individual peptide could be species-specific and dose-dependent. The relationships between peptides, locomotion and phagocytosis in these molluscs are discussed.     

Effect of consumption of green and black tea on the level of various enzymes in rats

Summary Drinking of both green and black tea as the only liquid ingested resulted in significant decreases in the activity of transketolase in whole blood of rats both before and after the in vitro addition, of thiamin diphosphate. Liver transketolase activity was decreased only by green tea. Mucosal transketolase activity was not affected by either type of tea. The activity of lactate dehydrogenase (LDH) was not affected by either type of tea, while whole blood LDH was decreased by both green and black tea. Neither tea had any affect on mucosal alkaline phosphatase, but thiamin diphosphatase activity was decreased by both teas. An increase in liver total thiamin resulted from the drinking of both types of tea.     

Evidence that the establishment of pregnancy requires activation of lipoxygenase and phospholipase-A2

Summary The present work investigates the possibility that lipoxygenase products are involved in the biochemical mechanisms of blastocyst implantation by utilizing nordihydroguaiaretic acid (NDGA) and caffeic acid (CA), inhibitors of lipoxygenase enzymes, and quinacrine (QU), an inhibitor of phospholipase-A₂. It has been shown previously that inhibition of cyclooxygenase results in blockade of implantation. The inhibitors were dissolved in a standard medium and 5 l of the solutions were micro-injected into the uterine horns of day-4 pregnant mice. The contralateral horns acted as controls and received only vehicle. A sham-operated group provided normal controls. In 14 NDGA-treated mice, the control horns contained 40 implantations while the treated horns contained only 6 small implantations and 8 resorbing sites. These control horns were comparable to the sham controls. In 14 CA-treated mice, treated horns contained 17 small implantations plus 4 resorptions, whereas the control horns contained 26 small implantations and 4 resorptions. Twelve QU-treated mice exhibited 7 small implantations and 4 resorptions in the treated horns, plus 24 small sites and no resorptions in the control horns. Fourteen sham-operated mice had 95 implantation sites and no resorptions in their 28 horns. The results provide evidence for the involvement of the lipoxygenase enzymes and phospholipase-A₂ in the initial implantation process and in the subsequent development of early pregnancy.     

Cellular models for the detection and evaluation of drugs that modulate human phagocyte activity

Summary Simple testing models have been developed for the evaluation of chemical or biological compounds that modulate the activity of human phagocytes. Human neutrophils from buffy coats of donor blood are used. They are stimulated with receptor agonists, and the effects of test compounds on exocytosis of different enzymes, the generation of superoxide (respiratory burst), and cytotoxicity are quantified. All assays are performed in microtiter plates and the responses are evaluated by multi-well photometry or fluorimetry. The models are apt to detect compounds acting on phagocytes as agonists or antagonists, signal transduction activators or inhibitors and primers of agonist responses, and to assess cytotoxic effects.     

Effects of chronic caffeine feeding on the activities of oxygen free radical defense enzymes in the growing rat heart and liver

The purpose of the present study was to determine the relationship between concentration of Zn, Cu and Fe, and the catalase, glutathione peroxidase and superoxide dismutase activities in the heart and liver of newborn rats whose dams were fed a diet supplemented with caffeine. Heart Zn levels of the 22- and 30-day-old rats of the caffeine group showed a decrease, whereas liver Zn levels showed an increase compared to the control. Cu levels in the liver at day 22 in the caffeine group were less than in the control. Cu- and Zn-containing superoxide dismutase activities showed an increase in the hearts of the caffeine group compared to the control. The activity of catalase and glutathion peroxidase showed no difference in the heart and liver between the groups. The present study suggests the possible involvement of superoxide dismutase enzyme in the impairment of heart formation as a result of chronic caffeine intake in the early growing period.     

Studies on some enzymes of the toad (Bufo melanostictus) testis and their probable role at the time of fertilization

Summary Glycosidases like sialidase,-galactosidase, -L-fucosidase, N-acetyl hexosaminidase and proteases were detected in toad testis. Neuraminic acid aldolase activity was also detected. The enzyme activities were found to vary as production of spermatozoa varied. All enzymes, except N-acetyl glucosaminidase, were shown to decrease after injection of toad pituitary extract and they were also found to be absent from testis containing no spermatozoa. The glycosidases were found to act on toad oviduct jelly and they may therefore be involved in the degradation of the jelly after fertilization, into smaller bits, which may be utilized as nutrients by the fertilized zygote.Acknowledgment. We thank Prof. T.R. Ramaiah, Head of the Department of Biochemistry, University of Mysore, for his help. We also acknowledge the financial assistance of University Grants Commission to one of us (MS) and CSIR through a grant No. 9 (165)83/EMR-II to HSS. Please address all correspondence to H.S. Seshadri.     

Novel cytotoxic peptides from the tropical marine cyanobacteriumHormothamnion enteromorphoides 1. Discovery,isolation and initial chemical and biological characterization of the hormothamnins from wild and cultured material

A Caribbean cyanobacterium,Hormothamnion enteromorphoides, was found to produce a complex mixture of ichthyotoxic peptides, perhaps explaining the apparent absence of predation upon these potentially palatable life forms. Bioassay-guided fractionation was used to isolate these toxic and antimicrobial natural products, and a variety of techniques including HR FAB mass spectrometry, 2D-NMR, traditional hydrolysis-amino acid analysis, and several chemical reactions were used to define the basic structural features of the major peptide, hormothamnin A. Hormothamnin A is a cyclic undecapeptide containing six common and five uncommon or new amino acid residues. HPLC analyses indicate that the relative proportions of these peptide natural products remain relatively constant between different collection locations and years, however, they do vary seasonally. Clonal isolates of this cyanobacterium in culture produce the full spectrum of toxic peptides.     

天宫一号目标飞行器载人环境设计与实施

基于天宫一号目标飞行器载人环境控制任务的新特点,以航天员的"舒适性"为设计目标,天宫一号进行了大气环境控制设计、噪声控制设计、辐射控制设计以及组合体热支持设计等载人环境设计,将组合体密封舱内的大气总压、温湿度、风速、气体成分、噪声以及辐射水平控制在人体舒适性范围内,天宫一号飞行任务表明,其载人环境系统设计正确,为飞行任务期间航天员的身体健康和工作效率提供有力保障.     

应急系统中数据收集与库设计

介绍应急系统中数据库设计思想、方法;分析了应急系统中所需的数据信息以及开发该应急系统所应建立的数据库;同时,还研究了应急系统中各数据库的相应属性及数据库之间所存在的联系。其目的是为了更好的收集、整理、共享处理应急突发事件时所需的各种数据信息,并将其以库的形式存储起来,为应急指挥部提供准确、具体的信息,为快速响应、协调行动提供信息基础。     

Theories that narrate the world: Ronald A. Fisher's mass selection and Sewall Wright's shifting balance

Theories are composed of multiple interacting components. I argue that some theories have narratives as essential components, and that narratives function as integrative devices of the mathematical components of theories. Narratives represent complex processes unfolding in time as a sequence of stages, and hold the mathematical elements together as pieces in the investigation of a given process. I present two case studies from population genetics: R. A. Fisher's “mas selection” theory, and Sewall Wright's shifting balance theory. I apply my analysis to an early episode of the “R. A. Fisher – Sewall Wright controversy.”     

印度科技政策与科技发展--兼对中印国家创新能力比较

印度科技政策对推动印度科技进步起着重要的促进作用,1958年“科学政策决议”奠定了基本原则,“2003年科学技术政策”明确了印度科技政策目标,还制订了科技战略行动计划。印度科技发展具有科技体制较为完善、科技投入不断增加、科研经费重国防轻民用、科技著作多、专利获得少、科技人才素质高、高科技异军突起等特点。与中国相比,印度国家创新能力评价更高。本文阐述了印度科技政策演变过程,分析了印度科技发展特征,论述了印度十五计划科技发展战略重点,并比较了中印国家创新能力。     

基于开放式的错误处理机制的设计与实现

本文探讨了软件项目中错误处理机制的作用及应用需求,结合软件项目开发中的实践经验,利用XML进行配置和描述,巧妙地设计与实现了一种通用的,基于开放式的错误处理机制,该机制配置灵活、操作方便、通用性好.     

Newtonianism and religion in the Netherlands

In the early eighteenth century Newtonianism became popular in the Netherlands both in academic (Boerhaave, ’sGravesande, Van Musschenbroek) and non-academic circles. The ‘Book of Nature’ was interpreted with the help of Newton’s natural philosophy and his ideas about a providential deity, thereby greatly enhancing the attractiveness of physico-theology in the eighteenth-century United Provinces. Like other Europeans the Dutch welcomed physico-theology as a strategic means in their battle against irreligion and atheism. Bernard Nieuwentijt, Johan Lulofs, Petrus Camper, and Johannes Florentius Martinet were prominent experts in the field. Combining Newtonian notions with Leibnizian optimism and romanticist trends, physico-theology remained popular in the Netherlands well into the nineteenth century.