Conservation of the sizes for one but not another class of exons in two chick collagen genes

Type III collagen is often found in the same tissues as type I collagen, yet the function and nature of the fibrils formed by the two collagens differ markedly. To understand the evolutionary history of the collagen gene family in more detail, we isolated the gene for type III collagen and compared its structure with that of the gene for alpha 2(I) collagen. This comparison points to a remarkable conservation in the size distribution of the exons coding for the helical part of these two collagen polypeptides: equivalent amino acid segments in the helical domain of each polypeptide are encoded by exons of equal sizes in each gene. This suggests that after the interstitial collagen genes had been duplicated from a common ancestor about 2-5 X 10(8) years ago, no recombinations between these exons were tolerated, although the same recombinational phenomena must have played an important part in shaping the structure of the progenitor for these genes. This fixation of the size distribution of the exons which code for the interstitial collagen helical domains is found despite the persistence in these exons of sequence elements that should have favoured recombinational rearrangements, and contrasts with the variations in the pattern of sizes of some exons coding for the amino and carboxyl propeptides of these collagens.     

Fibrils from brains of cows with new cattle disease contain scrapie-associated protein

During the past two years, more than 1,000 cases of a neurological disorder of cattle, bovine spongiform encephalopathy (BSE), have been confirmed from farms throughout Great Britain. The neurological signs and brain pathology of BSE resemble those produced in other species by the pathogens of scrapie and related disorders. The discovery of fibrils similar to scrapie-associated fibrils in detergent extracts o BSE-affected brain supported the clinical and pathological diagnosis of the disease, but has been controversial. Scrapie-associated fibrils are found in brain extracts of all species affected by scrapie and diseases caused by related pathogens. They are pathological aggregates of a neuronal membrane protein termed PrP and a protease-resistant form of PrP is a molecular marker of scrapie-associated fibrils. In this report, we show the major protein of BSE fibrils is the bovine homologue of PrP as judged by its size, protease resistance, immunoreactivity, lectin binding and partial N-terminal protein sequence. This confirms that BSE is a scrapie-like disease.     

Scrapie infectivity, fibrils and low molecular weight protein

The development of a short incubation model of scrapie (strain 263K), in golden hamsters has added impetus to the purification of the infectious agent. Our own attempts have been based on methods pioneered by Millson and developed by Prusiner. We present here results indicating that a purification factor of up to 10(4) with respect to protein may now be possible. Fractions from brain with high infectivity had a sedimentation range of 70-300S and contained an abundance of fibrils closely similar to the scrapie-associated fibrils (SAF) discovered by Merz et al.. Material of molecular weight (Mr) 26,000, which is probably protein, appears to be a major constituent of the fibrils. The association between infectivity and fibrils raises two possibilities: the fibrils are an infectious form of the scrapie agent or they are a pathological response to scrapie infection.     

胶原蛋白在LB膜上的吸附行为研究

基底的性质和制备方法对吸附胶原层的超分子自组装过程有很大的影响,将一种Gemini表面活性剂C18H37(CH3)2N+-(CH2)12-N+(CH3)2C18H37·2Br-(简写为18-12-18)利用LB技术在零压下转移到云母片上制成基底,浸入到pH值为3.0的胶原溶液中37℃恒温吸附不同的时间,样品用原子力显微镜(AFM)进行表征。结果表明:在恒温过程中,LB膜上的表面活性剂分子会发生移动和翻转,胶原分子优先吸附在LB膜上形成单层网状结构,随吸附时间的延长,胶原分子也会在云母上发生吸附,最后两者会交织成无序的胶原纤维。     

Structure of the cross-beta spine of amyloid-like fibrils

Numerous soluble proteins convert to insoluble amyloid-like fibrils that have common properties. Amyloid fibrils are associated with fatal diseases such as Alzheimer's, and amyloid-like fibrils can be formed in vitro. For the yeast protein Sup35, conversion to amyloid-like fibrils is associated with a transmissible infection akin to that caused by mammalian prions. A seven-residue peptide segment from Sup35 forms amyloid-like fibrils and closely related microcrystals, from which we have determined the atomic structure of the cross-beta spine. It is a double beta-sheet, with each sheet formed from parallel segments stacked in register. Side chains protruding from the two sheets form a dry, tightly self-complementing steric zipper, bonding the sheets. Within each sheet, every segment is bound to its two neighbouring segments through stacks of both backbone and side-chain hydrogen bonds. The structure illuminates the stability of amyloid fibrils, their self-seeding characteristic and their tendency to form polymorphic structures.     

Atomic structures of amyloid cross-beta spines reveal varied steric zippers

Amyloid fibrils formed from different proteins, each associated with a particular disease, contain a common cross-beta spine. The atomic architecture of a spine, from the fibril-forming segment GNNQQNY of the yeast prion protein Sup35, was recently revealed by X-ray microcrystallography. It is a pair of beta-sheets, with the facing side chains of the two sheets interdigitated in a dry 'steric zipper'. Here we report some 30 other segments from fibril-forming proteins that form amyloid-like fibrils, microcrystals, or usually both. These include segments from the Alzheimer's amyloid-beta and tau proteins, the PrP prion protein, insulin, islet amyloid polypeptide (IAPP), lysozyme, myoglobin, alpha-synuclein and beta(2)-microglobulin, suggesting that common structural features are shared by amyloid diseases at the molecular level. Structures of 13 of these microcrystals all reveal steric zippers, but with variations that expand the range of atomic architectures for amyloid-like fibrils and offer an atomic-level hypothesis for the basis of prion strains.     

Human blood platelets showing no response to collagen fail to express surface glycoprotein Ia

The interaction of blood platelets with collagen is generally considered to be of primary importance in the arrest of bleeding and to have a role in the pathogenesis of thrombosis and atherosclerosis. Following damage to the vascular endothelium, circulating platelets come into contact with exposed collagen fibrils in the subendothelium and spread along it; this is followed by the secretion of several biologically active substances and by aggregation of platelets. The glycoproteins of the platelet plasma membrane have an important role in the mechanisms underlying these processes. So far, two specific defects of platelet function in patients with a bleeding disorder are known to be associated with a glycoprotein defect and the study of these patients has contributed significantly to present concepts of platelet function. The glycoprotein (GP) IIB-III complex, absent or deleted in the aggregation-defective Glanzmann's thrombasthenia, has been identified as the platelet fibrinogen receptor. GPIb, which is absent in the adhesion-defective Bernard-Soulier syndrome, has been identified as the von Willebrand factor receptor on platelets. We now report a defect of the platelet plasma membrane glycoprotein composition in a patient whose platelets are totally unresponsive to collagen.     

Experimental manipulation of a contact guidance system in amphibian gastrulation by mechanical tension

Contact guidance has been implied in various morphogenetic movements including neural crest cell migration, primordial germ cell migration and guidance of axonal growth cone. In urodele gastrulae, we reported the presence of an aligned network of extracellular fibrils on the inside of the ectodermal layer and suggested that it guides the migration of the presumptive mesodermal cells from the blastopore towards the animal pole. We also reported in vitro experiments in which the fibril network of the ectodermal layer was transferred onto the surface of a coverslip. Dissociated mesodermal cells attach to and locomote actively on such conditioned surfaces in an oriented fashion along the blastopore-animal pole axis (bp-ap axis) of the ectodermal layer that conditioned the surface. Recent reports suggest that these fibrils contain fibronectin. We now report that the fibril network on the conditioned surface can be artificially aligned in any orientation by exerting mechanical tension on the ectodermal layer during the conditioning. Such prepared surfaces cause cell movements aligned along the tension axis, even when the tension axis is perpendicular to the natural axis of alignment along the bp-ap axis. These results suggest that the extracellular matrix fibrils aligned by the mechanical stress that arises in embryos during development can orient cell migration by the contact guidance, in a similar manner to that reported in the collagen gel and fibroblasts system.     

慢性不可预知性应激刺激建立大鼠颞下颌关节紊乱病模型

目的对大鼠进行人工干预下的情绪应激试验,观察大鼠下颌关节结构的变化,尝试建立颞下颌关节紊乱病(TMD)的动物模型。方法雄性Wistar大鼠16只,随机分为两组,分别为实验组和对照组。实验组进行慢性不可预知性应激试验,对照组正常饲养。观察各组大鼠行为学改变,6周后处死,完整解剖分离下颌髁状突及关节盘,扫描电镜观察。结果实验组大鼠行为学指标改变明显,扫描电镜下显示髁状突表面凝胶不完整,胶原纤维紊乱,断裂,暴露深层胶原组织等,关节盘表面凝胶样物质覆盖仍较完整,但沟回排列不规则,回隆起缩窄降低,沟变浅,对照组无此改变。结论慢性不可预见性应激刺激可造成大鼠下颌关节盘、突退行性改变,符合TMD病理改变,认为建立大鼠颞下颌关节紊乱病动物模型成功。     

Targeted pharmacological depletion of serum amyloid P component for treatment of human amyloidosis

The normal plasma protein serum amyloid P component (SAP) binds to fibrils in all types of amyloid deposits, and contributes to the pathogenesis of amyloidosis. In order to intervene in this process we have developed a drug, R-1-[6-[R-2-carboxy-pyrrolidin-1-yl]-6-oxo-hexanoyl]pyrrolidine-2-carboxylic acid, that is a competitive inhibitor of SAP binding to amyloid fibrils. This palindromic compound also crosslinks and dimerizes SAP molecules, leading to their very rapid clearance by the liver, and thus produces a marked depletion of circulating human SAP. This mechanism of drug action potently removes SAP from human amyloid deposits in the tissues and may provide a new therapeutic approach to both systemic amyloidosis and diseases associated with local amyloid, including Alzheimer's disease and type 2 diabetes.     

The most infectious prion protein particles

Neurodegenerative diseases such as Alzheimer's, Parkinson's and the transmissible spongiform encephalopathies (TSEs) are characterized by abnormal protein deposits, often with large amyloid fibrils. However, questions have arisen as to whether such fibrils or smaller subfibrillar oligomers are the prime causes of disease. Abnormal deposits in TSEs are rich in PrP(res), a protease-resistant form of the PrP protein with the ability to convert the normal, protease-sensitive form of the protein (PrP(sen)) into PrP(res) (ref. 3). TSEs can be transmitted between organisms by an enigmatic agent (prion) that contains PrP(res) (refs 4 and 5). To evaluate systematically the relationship between infectivity, converting activity and the size of various PrP(res)-containing aggregates, PrP(res) was partially disaggregated, fractionated by size and analysed by light scattering and non-denaturing gel electrophoresis. Our analyses revealed that with respect to PrP content, infectivity and converting activity peaked markedly in 17-27-nm (300-600 kDa) particles, whereas these activities were substantially lower in large fibrils and virtually absent in oligomers of < or =5 PrP molecules. These results suggest that non-fibrillar particles, with masses equivalent to 14-28 PrP molecules, are the most efficient initiators of TSE disease.     

Conformational change of glutathione-S-transferase by its co-expression with prion domain of yeast Ure2p

Ure2 protein from Saccharomyces cerevisisae has a changeable structure similar to that ofrnammalian prion protein. Its N-terminal is the prion domain (PrD) consisting of 65 amino acids which plays a critical role in yeast prion development. In this study, PrD gene was recombinated with glutathione-S-transferase(GST) gene, and a soluble GST-PrD(sGST-PrD) fusion protein was expressed in E. coli. sGST-PrD could spontaneously polymerize into amyloid fibrils in vitro, displaying typical β-sheet-type structure; it had increased resistance to proteinase K and exhibited amvloid-like optical properties. Moreover, the aggregated GST-PrD(aGST-PrD) could induce sGST-PrD to aggregate into fibrils. These results indicate that PrD could change the conformation of GST moiety in a recombinant protein with PrD to form a prion-like chimeric protein, which proves that PrD has the ability to mediate a prion-like conversion of other proteins fused with it.     

番木瓜、萝卜和油菜可溶性和结合型芥子酶活性的比较

芥子酶在植物细胞中有可溶性的自由二聚体和不溶的结合形式两种存在形态。本研究发现物种间芥子酶的形态有较大差异。番木瓜芥子酶在种子、子叶和真叶等3种组织中,主要以可溶形式存在,不溶态芥子酶微量。而在油菜种子中,芥子酶主要以结合形式存在,在子叶和真叶中,芥子酶则主要以可溶形式存在。萝卜的芥子酶介于二者之间。供试3个物种的可溶性和不溶性芥子酶都存在相互抑制的现象。     

葛仙米及其生长土壤的扫描电子显微镜观察

借助扫描电子显微镜对鹤峰走马葛仙米及对应的土壤样品进行检测,发现葛仙米的表层发育出2条胶质的薄层长丝体;同时葛仙米表层有一个孔.从土壤样品可观察到葛仙米母体解体后呈胶质的鞘丝体和胶质的片断.葛仙米对土壤颗粒具有集结粘连的作用.     

Biomimetic self-templating supramolecular structures

In nature, helical macromolecules such as collagen, chitin and cellulose are critical to the morphogenesis and functionality of various hierarchically structured materials. During tissue formation, these chiral macromolecules are secreted and undergo self-templating assembly, a process whereby multiple kinetic factors influence the assembly of the incoming building blocks to produce non-equilibrium structures. A single macromolecule can form diverse functional structures when self-templated under different conditions. Collagen type I, for instance, forms transparent corneal tissues from orthogonally aligned nematic fibres, distinctively coloured skin tissues from cholesteric phase fibre bundles, and mineralized tissues from hierarchically organized fibres. Nature's self-templated materials surpass the functional and structural complexity achievable by current top-down and bottom-up fabrication methods. However, self-templating has not been thoroughly explored for engineering synthetic materials. Here we demonstrate the biomimetic, self-templating assembly of chiral colloidal particles (M13 phage) into functional materials. A single-step process produces long-range-ordered, supramolecular films showing multiple levels of hierarchical organization and helical twist. Three distinct supramolecular structures are created by this approach: nematic orthogonal twists, cholesteric helical ribbons and smectic helicolidal nanofilaments. Both chiral liquid crystalline phase transitions and competing interfacial forces at the interface are found to be critical factors in determining the morphology of the templated structures during assembly. The resulting materials show distinctive optical and photonic properties, functioning as chiral reflector/filters and structural colour matrices. In addition, M13 phages with genetically incorporated bioactive peptide ligands direct both soft and hard tissue growth in a hierarchically organized manner. Our assembly approach provides insight into the complexities of hierarchical assembly in nature and could be expanded to other chiral molecules to engineer sophisticated functional helical-twisted structures.     

木棉纤维的微细结构研究——胞壁层次结构与原纤尺度

通过对木棉纤维横截面超薄切片的透射电镜观察，获得了木棉纤维的胞壁层次结构、原纤尺度及排列。木棉纤维胞壁具有清晰可见的多层状结构，基本上可以分为外表皮层S、胞壁1层W1、胞壁2层W2、胞壁3层W3和内皮层IS共5个基本的层次，各层具有不同的厚度和原纤堆砌密度与排列方向。可见木棉纤维横截面最小结构单元宽度为3．2～5．0nm，与棉纤维基原纤尺寸相当。实验证明，碱液对木棉纤维的可及性存在显著的个体差异；对于同一纤维胞壁各层的溶胀也存在差异。其中W2是最易被溶胀的；内皮层的原纤容易被分离出来。而未经溶胀处理的木棉纤维电镜照片反差弱，层次结构不够细致。     

Target size regulates calibre and myelination of sympathetic axons

Axons in vertebrate peripheral nerves are ensheathed by Schwann cells. For some axons, this sheath consists of a single layer of glial cell cytoplasm and plasma membranes; for other axons, Schwann cells form multilayered myelin. Whether or not a Schwann cell makes myelin is determined by a signal from the axon, but the nature of this signal is not known. Here I show that sympathetic postganglionic axons, which are normally not myelinated, become myelinated when their calibre is increased as a result of increasing the size of the peripheral target they innervate. This result implies that axon calibre, which is known to be correlated with myelination, is in fact the crucial determinant of whether an axon becomes myelinated. Furthermore, the finding that increasing or decreasing target size causes corresponding increases or decreases in axon size indicates that axon calibre is itself regulated by retrograde signals from peripheral target tissues.     

基于活塞挤出的组织工程支架低温沉积制造工艺

快速成形(RP)工艺可实现个性化制造和可控的材料、结构组成,因此在组织工程中得到广泛应用。该文设计开发基于活塞挤出的材料喷头,对挤出工艺进行探讨与研究,提出采用压力自释放方案解决其流涎问题,并将其应用于低温沉积制造(LDM)工艺。实验表明,基于活塞挤出的LDM工艺可以成形组织工程支架,并且具有广泛的材料适应性,可成形多种生物材料,尤其可成形其他RP工艺难于成形的胶原材料。     

共同性斜视眼肌的组织学研究

用光镜和电镜研究１１例共同性斜视患者手术切除的眼肌，结果表明：横纹肌细胞发生进行性变性坏死，胶原纤维增生。与此同时，神经纤维减少甚至消失，说明共同性斜视眼肌和神经纤维的确产生病变，这些病变可能与感染有关。     

共同性斜视眼肌的组织学研究

用光镜和电镜研究１１例共同性斜视患者手术切除的眼肌，结果表明，横 肌细胞发生进行性变性坏死，胶原纤维增生，与此同时，神经纤维减少甚至消失，说明共同性斜视眼肌和神经纤维的确产生病变，这些病变可能与感染有关。