Studies on the regioselectivity and kinetics of the action of trypsin on proinsulin and its derivatives using mass spectrometry

Human M-proinsulin was cleaved by trypsin at the R³¹R³²–E³³ and K⁶⁴R⁶⁵–G⁶⁶ bonds (B/C and C/A junctions), showing the same cleavage specificity as exhibited by prohormone convertases 1 and 2 respectively. Buffalo/bovine M-proinsulin was also cleaved by trypsin at the K⁵⁹R⁶⁰–G⁶¹ bond but at the B/C junction cleavage occurred at the R³¹R³²–E³³ as well as the R³¹–R³²E³³ bond. Thus, the human isoform in the native state, with a 31 residue connecting C-peptide, seems to have a unique structure around the B/C and C/A junctions and cleavage at these sites is predominantly governed by the structure of the proinsulin itself. In the case of both the proinsulin species the cleavage at the B/C junction was preferred (65%) over that at the C/A junction (35%) supporting the earlier suggestion of the presence of some form of secondary structure at the C/A junction. Proinsulin and its derivatives, as natural substrates for trypsin, were used and mass spectrometric analysis showed that the k_cat./K_m values for the cleavage were most favourable for the scission of the bonds at the two junctions (1.02 ± 0.08 × 10⁵ s^− 1 M^− 1) and the cleavage of the K²⁹–T³⁰ bond of M-insulin-RR (1.3 ± 0.07 × 10⁵ s^− 1 M^− 1). However, the K²⁹–T³⁰ bond in M-insulin, insulin as well as M-proinsulin was shielded from attack by trypsin (k_cat./K_m values around 1000 s^− 1 M^− 1). Hence, as the biosynthetic path follows the sequence; proinsulin → insulin-RR → insulin, the K²⁹–T³⁰ bond becomes shielded, exposed then shielded again respectively.     

Analysis of the genetic effects for several traits in oil palm (Elaeis guineensis Jacq.) populations. I. Population means

An oil palm experiment was set up in the Ivory Coast to compare the effects of crossing and selfing within two origins, Deli and La Mé, on the mean and the variability of Deli x Mé between-origin hybrids. The originality of the experiment lay in the crossing plan, which provided access to genetic parameters related to additivity, dominance and different components of epistasis. This first part covers the analysis of the components of the mean. The parents used were obtained from four palms, two from each origin. Those of La Mé origin were half-sibs. The common parent came from a wild stand in the Ivory Coast. Those of Deli origin were from two different populations bred in Southeast Asia for several generations from a narrow genetic base. These four parents gave rise to nine Deli x La Mé hybrid populations with double-cousintype links. The additive component is more important within the Deli origin than within the La Mé origin. This may be explained by the large genetic divergence between the two Deli parent palms. On the other hand, the additive^* additive epistasis is more substantial within the La Mé origin, probably because of inbreeding. The discussion concentrates on how this information should be used when choosing parents to be crossed and tested and to produce improved populations. The crossing plan proposed can be of general use and is suitable for other species in a reciprocal recurrent selection programme.     

基于GIS的视觉景观影响定量评价方法理论与实践

在人地关系日益紧张的今天,一些可能产生视觉景观不良影响的项目不得不建设于自然保护区、历史文化遗迹等视觉景观敏感区内或附近,为避免建设项目对自然、历史、文化、旅游资源产生不必要的损失或造成难以弥补的遗憾,视觉景观环境影响评价就显得尤为重要.视觉景观环境影响评价在中国尚处于探索阶段,相关的研究多数集中在理论综述和定性的评价.以某建设项目对明十三陵景区的景观影响评价为例,对如何运用GIS空间分析功能,开展视觉景观环境影响定量评价研究展开探讨.     

Triterpenoid saponins from Fatsia japonica

Five triterpenoid saponins isolated from the flowers, the mature fruits and the leaves of Fatsia japonica were identified as 3-O-[β-d-glucopyranosyl(1→4)-β-d-glucopyranosyl]-hederagenin (1), 3-O-[β-d-glucopyranosyl-(1→4)-α-l-arabinopyranosyl]-oleanolic acid (2), 3-O-[α-l-arabinopyranosyl]-hederagenin (3), 3-O-[β-d-glucopyranosyl]-hederagenin (4) and 3-O-[β-d-glucopyranosyl(1→4)-α-l-arabinopyranosyl]-hederagenin (5). The saponins 1 and 2 are new, naturally occurring, triterpenoid saponins. The distribution of the five saponins in three parts of the plant was investigated. Saponins 2, 3 and 5 were present in the flowers, saponins 1, 3, 4 and 5 were in the mature fruits and saponins 2, 3, 4 and 5 were in the leaves.     

苏北沿海滩涂养殖湿地磷化氢的释放及其影响因素

利用静态暗箱采样方法和气相色谱-氮磷检测器(GC-NPD)分析技术测定了苏北沿海滩涂养殖湿地区域磷化氢的释放通量,研究了其季节和空间变化特征,并探讨了其潜在的影响因素。结果表明,各季节磷化氢释放通量的高低排序为:8月份5月份11月份2月份,其变化范围分别为16.5—168.9、8.3—105.9、8.2—64.5和-19.5—49.6 ng m-2h-1,平均值分别为65.0、36.5、25.9和16.4 ng m-2h-1;空间变化明显,各季节的平均值,b10站位最高,b3次之,b5和b8站位相对较低。磷化氢释放通量与沉积环境因子的相关性分析显示,释放通量与TP、IP、OP、OC、TN、SC和T有较好的线性正相关关系(相关系数R分别为0.807、0.579、0.828、0.825、0.467、0.605和0.551,P值0.01),与Eh有较好的线性负相关关系(R为-0.774,P值0.01),表明在研究调查区域,较高的磷、碳、氮、硫组分含量,较高的温度和较低的氧化还原电位更有利于磷化氢的释放过程。     

Differential susceptibility to hydrogen sulfide-induced apoptosis between PHLDA1-overexpressing oral cancer cell lines and oral keratinocytes: Role of PHLDA1 as an apoptosis suppressor

Hydrogen sulfide (H₂S) is a novel gasotransmitter that plays multiple biological roles in various body systems. In addition to its endogenous production, H₂S is produced by bacteria colonizing digestive organs, including the oral cavity. H₂S was previously shown to enhance pro-apoptotic effects in cancer cell lines, although the mechanisms involved remain unclear. To properly assess the anti-cancer effects of H₂S, however, investigations of apoptotic effects in normal cells are also necessary. The aims of this study were (1) to compare the susceptibility to H₂S-induced apoptosis between the oral cancer cell line Ca9-22 and oral keratinocytes that were derived from healthy gingiva, and (2) to identify candidate genes involved in the induction of apoptosis by H₂S. The susceptibility to H₂S-induced apoptosis in Ca9-22 cells was significantly higher than that in keratinocytes. H₂S exposure in Ca9-22 cells, but not keratinocytes, enhanced the expression of pleckstrin homology-like domain, family A, member 1 (PHLDA1), which was identified through a differential display method. In addition, PHLDA1 expression increased during actinomycin D-induced apoptosis in Ca9-22 cells. Knockdown of PHLDA1 expression by small interfering RNA in Ca9-22 cells led to expression of active caspase 3, thus indicating apoptosis induction. The tongue cancer cell line SCC-25, which expresses PHLDA1 at a high level, showed similar effects. Our data indicate that H₂S is an anti-cancer compound that may contribute to the low incidence of oral cancer. Furthermore, we demonstrated the role of PHLDA1 as an apoptosis suppressor.     

HisE11 and HisF8 Provide Bis-histidyl Heme Hexa-coordination in the Globin Domain of Geobacter sulfurreducens Globin-coupled Sensor

Among heme-based sensors, recent phylogenomic and sequence analyses have identified 34 globin coupled sensors (GCS), to which an aerotactic or gene-regulating function has been tentatively ascribed. Here, the structural and biochemical characterization of the globin domain of the GCS from Geobacter sulfurreducens (GsGCS¹⁶²) is reported. A combination of X-ray crystallography (crystal structure at 1.5 Å resolution), UV-vis and resonance Raman spectroscopy reveals the ferric GsGCS¹⁶² as an example of bis-histidyl hexa-coordinated GCS. In contrast to the known hexa-coordinated globins, the distal heme-coordination in ferric GsGCS¹⁶² is provided by a His residue unexpectedly located at the E11 topological site. Furthermore, UV-vis and resonance Raman spectroscopy indicated that ferrous deoxygenated GsGCS¹⁶² is a penta-/hexa-coordinated mixture, and the heme hexa-to-penta-coordination transition does not represent a rate-limiting step for carbonylation kinetics. Lastly, electron paramagnetic resonance indicates that ferrous nitrosylated GsGCS¹⁶² is a penta-coordinated species, where the proximal HisF8-Fe bond is severed.     

The D' measure of overall gametic disequilibrium between pairs of multiallelic loci

Abstract The D ' coefficient is one of the most commonly used measures of the extent of gametic disequilibrium between multiallelic loci. It has been suggested that the range of the D ' measure of overall disequilibrium between pairs of multiallelic loci depends on allele frequencies, except under some very restricted conditions. Nevertheless, the problem of dependence of the range of D ' has not been characterized under a wide set of possible polymorphisms. Evaluation of the utility of D ' as a measure of the strength of overall disequilibrium between all possible pairs of alleles at two multiallelic loci requires better knowledge of its range than is currently available. In this work, the conditions of polymorphism under which the range of D ' is frequency independent are given. It is found that the range of D ' is more often independent of allelic frequencies than is commonly thought. Furthermore, the range of D ' undergoes only small fluctuations as a function of the polymorphisms at the loci. Numerical cases and microsatellite data from humans are used for illustration. These observations indicate that the D ' coefficient is a useful tool for the estimation and comparison of the extent of overall disequilibrium across pairs of multiallelic loci.     

Virtual resource development in the glycosciences

The development of Internet-based virtual resources is a relatively new area of scientific and technical activity that is currently undergoing rapid expansion. Major factors fuelling recent growth include the emergence of multimedia capabilities through the rapid evolution of the World Wide Web, the reduction in cost of high quality personal computers and graphics workstations and the provision of mass-marketed provider services. Prior to 1995 the presence of Internet resources in the glycosciences was virtually non-existent. Existing scientific knowledge was primarily made available on the Net through the provision of databases from gopher and ftp sites. A particular example in the glycosciences is the Carbbank database of biological carbohydrate sequences. We will describe here our efforts in 1994–95 in establishing The Glycoscience Network (TGN, http://bellatrix.pcl.ox.ac.uk/TGN/). These activities included the establishment of a newsgroup, mailing lists, Web resources and the running of the First Electronic Glycoscience Conference (EGC-1, http://bellatrix.pcl.ox.ac.uk/egc/). EGC-1 included many novel initiatives in the glycosciences including electronic posters and papers, a Virtual Conference Centre, a Web-based hyperglossary, Virtual Trade and Employment Centres, refereed electronic publishing, and the creation of a Virtual Reality Gallery. We would like to look towards the near future and discuss several initiatives in virtual resource creation that we believe will have significant scientific impact on the glycosciences including the development of bioinformatics-based servers, sophisticated interactive databases, and videoconferencing. Furthermore, we cherish the belief that these resources will foster international scientific collaboration and progress of an extent never previously possible. Finally, we indulge in speculation and make some suggestions on the form and long-term impact of Glycoscience Virtual Resources. We predict that their development may completely reconstruct the scientific environment that we work in as scientists and we reflect on the probable benefits and pitfalls to be encountered.This paper was presented at the First Electronic Glycoscience Conference (EGCI) on the World Wide Web, September 1995.