Proteases of Antarctic krill--a new system for effective enzymatic debridement of necrotic ulcerations

The Antarctic krill (Euphausia superba) possesses an "over-dimensioned' digestive system, which is of vital importance for the survival of this euphaucean shrimp in the extreme marine environment. The isolated enzymes contain a well-balanced mixture of both endo- and exopeptidases, assuring fast and complete breakdown of proteinaceous material. These unique properties have now been shown to be extremely valuable for the effective removal of necrotic debris, fibrin or blood crusts in vitro. Therefore the krill enzymes should be considered as an important resource in the future management of necrotic wounds.     

Serum dopamine-beta-hydroxylase activity in a new strain of spontaneously hypertensive rats

Summary The serum dopamine--hydroxylase (DBH) activity is higher in 5-week-old rats of the Lyon Hypertensive strain than in rats of the Lyon Normotensive strain. This difference disappears in older animals when the hypertension is developed, while the DBH activity decreases in the both strains.This work was supported by grants from DGRST (77.7.1930), CNRS (RCP 080.469, ATP A651-3358), INSERM (15.75.38) and the Fondation pour la Recherche Médicale Française.     

社鼠和针毛鼠线粒体DNA序列的歧异及其系统进化关系

社鼠和针毛鼠是有着广泛地理分布的山地鼠种,它们的一些形态特征及核型明显有别于同属的其它鼠类.社鼠的亚种--雷琼社鼠局限于分布在广东省西部地区和海南省,它的形态特征多似社鼠,但也有少数特征相似于针毛鼠.对于社鼠和针毛鼠的分类地位及社鼠亚种的分化一直有着各种讨论.本研究进行了社鼠和雷琼社鼠以及针毛鼠龙门种群和香港种群的细胞色素b (264个核苷酸位点)和12S rRNA (387个核苷酸位点)基因片段的测序.社鼠与雷琼社鼠线粒体的细胞色素b基因片段间包含有19个变异位点、12S rRNA 基因片段间包含11个变异位点和2个插入/缺失位点.针毛鼠的2个种群间的细胞色素b 和12S rRNA 基因片段分别包含了3个和11个变异位点.通过邻接法和最大简约法重建的系统进化关系表明:雷琼社鼠与社鼠有着紧密的亲缘关系,它的亚种地位得到了线粒体DNA序列证据的肯定.针毛鼠龙门种群和香港种群细胞色素b和12S rRNA 基因片段序列的歧异反映出两个种群由于长期的地理隔离及其所经历自然选择的结果.     

The mechanism of sodium and substrate release from the binding pocket of vSGLT

Membrane co-transport proteins that use a five-helix inverted repeat motif have recently emerged as one of the largest structural classes of secondary active transporters. However, despite many structural advances there is no clear evidence of how ion and substrate transport are coupled. Here we report a comprehensive study of the sodium/galactose transporter from Vibrio parahaemolyticus (vSGLT), consisting of molecular dynamics simulations, biochemical characterization and a new crystal structure of the inward-open conformation at a resolution of 2.7??. Our data show that sodium exit causes a reorientation of transmembrane helix 1 that opens an inner gate required for substrate exit, and also triggers minor rigid-body movements in two sets of transmembrane helical bundles. This cascade of events, initiated by sodium release, ensures proper timing of ion and substrate release. Once set in motion, these molecular changes weaken substrate binding to the transporter and allow galactose readily to enter the intracellular space. Additionally, we identify an allosteric pathway between the sodium-binding sites, the unwound portion of transmembrane helix 1 and the substrate-binding site that is essential in the coupling of co-transport.     

Rapid electroplating of insulators

Electrochemical techniques for depositing metal films and coatings have a long history. Such techniques essentially fall into two categories, with different advantages and disadvantages. The first, and oldest, makes use of spontaneous redox reactions to deposit a metal from solution, and can be used on both insulating and metallic substrates. But the deposition conditions of these processes are difficult to control in situ, in part because of the variety of salts and additives present in the solution. The second approach-electroplating-uses an electric current to reduce metal ions in solution, and offers control over the quantity (and, to some extent, grain size) of deposited metal. But application of this technique has hitherto been restricted to conducting substrates. Here we describe an electroplating technique that permits coating of insulating substrates with metals having controlled grain size, thickness and growth speed. The basis of our approach is the progressive outward growth of the metal from an electrode in contact with the substrate, with the cell geometry chosen so that the electron current providing the reduction passes through the growing deposit. Such an approach would normally form dendritic or powdery deposits, but we identify a range of conditions in which uniform films rapidly form.     

FACL4, encoding fatty acid-CoA ligase 4, is mutated in nonspecific X-linked mental retardation

X-linked mental retardation (XLMR) is an inherited condition that causes failure to develop cognitive abilities, owing to mutations in a gene on the X chromosome. The latest XLMR update lists up to 136 conditions leading to 'syndromic', or 'specific', mental retardation (MRXS) and 66 entries leading to 'nonspecific' mental retardation (MRX). For 9 of the 66 MRX entries, the causative gene has been identified. Our recent discovery of the contiguous gene deletion syndrome ATS-MR (previously known as Alport syndrome, mental retardation, midface hypoplasia, elliptocytosis, OMIM #300194), characterized by Alport syndrome (ATS) and mental retardation (MR), indicated Xq22.3 as a region containing one mental retardation gene. Comparing the extent of deletion between individuals with ATS-MR and individuals with ATS alone allowed us to define a critical region for mental retardation of approximately 380 kb, containing four genes. Here we report the identification of two point mutations, one missense and one splice-site change, in the gene FACL4 in two families with nonspecific mental retardation. Analysis of enzymatic activity in lymphoblastoid cell lines from affected individuals of both families revealed low levels compared with normal cells, indicating that both mutations are null mutations. All carrier females with either point mutations or genomic deletions in FACL4 showed a completely skewed X-inactivation, suggesting that the gene influences survival advantage. FACL4 is the first gene shown to be involved in nonspecific mental retardation and fatty-acid metabolism.     

Role of transposable elements in heterochromatin and epigenetic control

Heterochromatin has been defined as deeply staining chromosomal material that remains condensed in interphase, whereas euchromatin undergoes de-condensation. Heterochromatin is found near centromeres and telomeres, but interstitial sites of heterochromatin (knobs) are common in plant genomes and were first described in maize. These regions are repetitive and late-replicating. In Drosophila, heterochromatin influences gene expression, a heterochromatin phenomenon called position effect variegation. Similarities between position effect variegation in Drosophila and gene silencing in maize mediated by "controlling elements" (that is, transposable elements) led in part to the proposal that heterochromatin is composed of transposable elements, and that such elements scattered throughout the genome might regulate development. Using microarray analysis, we show that heterochromatin in Arabidopsis is determined by transposable elements and related tandem repeats, under the control of the chromatin remodelling ATPase DDM1 (Decrease in DNA Methylation 1). Small interfering RNAs (siRNAs) correspond to these sequences, suggesting a role in guiding DDM1. We also show that transposable elements can regulate genes epigenetically, but only when inserted within or very close to them. This probably accounts for the regulation by DDM1 and the DNA methyltransferase MET1 of the euchromatic, imprinted gene FWA, as its promoter is provided by transposable-element-derived tandem repeats that are associated with siRNAs.     

Mutant small heat-shock protein 27 causes axonal Charcot-Marie-Tooth disease and distal hereditary motor neuropathy

Charcot-Marie-Tooth disease (CMT) is the most common inherited neuromuscular disease and is characterized by considerable clinical and genetic heterogeneity. We previously reported a Russian family with autosomal dominant axonal CMT and assigned the locus underlying the disease (CMT2F; OMIM 606595) to chromosome 7q11-q21 (ref. 2). Here we report a missense mutation in the gene encoding 27-kDa small heat-shock protein B1 (HSPB1, also called HSP27) that segregates in the family with CMT2F. Screening for mutations in HSPB1 in 301 individuals with CMT and 115 individuals with distal hereditary motor neuropathies (distal HMNs) confirmed the previously observed mutation and identified four additional missense mutations. We observed the additional HSPB1 mutations in four families with distal HMN and in one individual with CMT neuropathy. Four mutations are located in the Hsp20-alpha-crystallin domain, and one mutation is in the C-terminal part of the HSP27 protein. Neuronal cells transfected with mutated HSPB1 were less viable than cells expressing the wild-type protein. Cotransfection of neurofilament light chain (NEFL) and mutant HSPB1 resulted in altered neurofilament assembly in cells devoid of cytoplasmic intermediate filaments.     

A previously unidentified MECP2 open reading frame defines a new protein isoform relevant to Rett syndrome

Rett syndrome is caused by mutations in the gene MECP2 in approximately 80% of affected individuals. We describe a previously unknown MeCP2 isoform. Mutations unique to this isoform and the absence, until now, of identified mutations specific to the previously recognized protein indicate an important role for the newly discovered molecule in the pathogenesis of Rett syndrome.