SynGAP isoforms exert opposing effects on synaptic strength.

Alternative promoter usage and alternative splicing enable diversification of the transcriptome. Here we demonstrate that the function of Synaptic GTPase-Activating Protein (SynGAP), a key synaptic protein, is determined by the combination of its amino-terminal sequence with its carboxy-terminal sequence. 5' rapid amplification of cDNA ends and primer extension show that different N-terminal protein sequences arise through alternative promoter usage that are regulated by synaptic activity and postnatal age. Heterogeneity in C-terminal protein sequence arises through alternative splicing. Overexpression of SynGAP α1 versus α2 C-termini-containing proteins in hippocampal neurons has opposing effects on synaptic strength, decreasing and increasing miniature excitatory synaptic currents amplitude/frequency, respectively. The magnitude of this C-terminal-dependent effect is modulated by the N-terminal peptide sequence. This is the first demonstration that activity-dependent alternative promoter usage can change the function of a synaptic protein at excitatory synapses. Furthermore, the direction and degree of synaptic modulation exerted by different protein isoforms from a single gene locus is dependent on the combination of differential promoter usage and alternative splicing.

鲢(Hypophthalmichthys molitrix)IL-10基因的克隆及表达分析

白细胞介素10(IL-10)是一种作用广泛的抗炎细胞因子,主要功能是限制和最终终止炎症反应．用RACE(rapid amplification of cDNA ends)-PCR方法扩增出鲢白细胞介素10(IL-10)的 cDNA,其全长为1248nt,包含5’非编码区156 nt,3’非编码区552nt,开放阅读框540nt．鲢IL- 10的开放阅读框编码179个氨基酸,其中包含构成两对二硫键的4个保守半胱氨酸．RT-PCR结果显示鲢IL-10 mRNA主要在脾脏、鳃、头肾和肌肉中表达．将鲢IL-10完

Blade Loading and Its Application in the Mean-Line Design of Low Pressure Turbines

In order to minimize the number of iterations to a turbine design, reasonable choices of the key parameters must be made at the preliminary design stage. The choice of blade loading is of particular concern in the low pressure (LP) turbine of civil aero engines, where the use of high-lift blades is widespread. This paper considers how blade loading should be measured, compares the performance of various loss correlations, and explores the impact of blade lift on performance and lapse rates. To these ends, an analytical design study is presented for a repeating-stage, axial-flow LP turbine. It is demonstrated that the long-established Zweifel lift coefficient (Zweifel, 1945, "The Spacing of Turbomachine Blading, Especially with Large Angular Deflection" Brown Boveri Rev., 32(1), pp. 436-444) is flawed because it does not account for the blade camber. As a result the Zweifel coefficient is only meaningful for a fixed set of flow angles and cannot be used as an absolute measure of blade loading. A lift coefficient based on circulation is instead proposed that accounts for the blade curvature and is independent of the flow angles. Various existing profile and secondary loss correlations are examined for their suitability to preliminary design. A largely qualitative comparison demonstrates that the loss correlations based on Ainley and Mathieson (Ainley and Mathieson, 1957, "A Method of Performance Estimation for Axial-Flow Turbines," ARC Reports and Memoranda No. 2974; Dunham and Came, 1970, "Improvements to the Ainley-Mathieson Method of Turbine Performance Prediction," Trans. ASME: J. Eng. Gas Turbines Power, July, pp. 252-256; Kacker and Okapuu, 1982, "A Mean Line Performance Method for Axial Flow Turbine Efficiency," J. Eng. Power, 104, pp. 111-119). are not realistic, while the profile loss model of Coull and Hodson (Coull and Hodson, 2011, "Predicting the Profile Loss of High-Lift Low Pressure Turbines," J. Turbomach., 134(2), pp. 021002) and the secondary loss model of (Traupel, W, 1977, Thermische Turbomaschinen, Springer-Verlag, Berlin) are arguably the most reasonable. A quantitative comparison with multistage rig data indicates that, together, these methods over-predict lapse rates by around 30%, highlighting the need for improved loss models and a better understanding of the multistage environment. By examining the influence of blade lift across the Smith efficiency chart, the analysis demonstrates that designs with higher flow turning will tend to be less sensitive to increases in blade loading. © 2013 American Society of Mechanical Engineers.

Structurally programmed capillary folding of carbon nanotube assemblies.

We demonstrate the fabrication of horizontally aligned carbon nanotube (HA-CNT) networks by spatially programmable folding, which is induced by self-directed liquid infiltration of vertical CNTs. Folding is caused by a capillary buckling instability and is predicted by the elastocapillary buckling height, which scales with the wall thickness as t(3/2). The folding direction is controlled by incorporating folding initiators at the ends of the CNT walls, and the initiators cause a tilt during densification which precedes buckling. By patterning these initiators and specifying the wall geometry, we control the dimensions of HA-CNT patches over 2 orders of magnitude and realize multilayered and multidirectional assemblies. Multidirectional HA-CNT patterns are building blocks for custom design of nanotextured surfaces and flexible circuits.

The soft impact response of composite laminate beams

The quasi-static and dynamic responses of laminated beams of equal areal mass, made from monolithic CFRP and Ultra high molecular weight Polyethylene (UHMWPE), have been measured. The end-clamped beams were impacted at mid-span by metal foam projectiles to simulate localised blast loading. The effect of clamping geometry on the response was investigated by comparing the response of beams bolted into the supports with the response of beams whose ends were wrapped around the supports. The effect of laminate shear strength upon the static and dynamic responses was investigated by testing two grades of each of the CFRP and UHMWPE beams: (i) CFRP beams with a cured matrix and uncured matrix, and (ii) UHMWPE laminates with matrices of two different shear strengths. Quasi-static stretch-bend tests indicated that the load carrying capacity of the UHWMPE beams exceeds that of the CFRP beams, increases with diminishing shear strength of matrix, and increases when the ends are wrapped rather than through-bolted. The dynamic deformation mode of the beams is qualitatively different from that observed in the quasi-static stretch-bend tests. In the dynamic case, travelling hinges emanate from the impact location and propagate towards the supports; the beams finally fail by tensile fibre fracture at the supports. The UHMWPE beams outperform the CFRP beams in terms of a lower mid-span deflection for a given impulse, and a higher failure impulse. Also, the maximum attainable impulse increases with decreasing shear strength for both the UHMWPE and CFRP beams. The ranking of the beams for load carrying capacity in the quasi-static stretch-bend tests is identical to that for failure impulse in the impact tests. Thus, the static tests can be used to gauge the relative dynamic performances of the beams. © 2013 Elsevier Ltd. All rights reserved.

Simulations of thermal upgrading methods for oil shale reservoirs

This paper analyzes reaction and thermal front development in porous reservoirs with reacting flows, such as those encountered in shale oil extraction. A set of dimensionless parameters and a 3D code are developed in order to investigate the important physical and chemical variables of such reservoirs when heated by in situ methods. This contribution builds on a 1D model developed for the precursor study to this work. Theory necessary for this study is presented, namely shale decomposition chemical mechanisms, governing equations for multiphase flow in porous media and necessary closure models. Plotting the ratio of the thermal wave speed to the fluid speed allows one to infer that the reaction wave front ends where this ratio is at a minimum. The reaction front follows the thermal front closely, thus allowing assumptions to be made about the extent of decomposition solely by looking at thermal wave progression. Furthermore, this sensitivity analysis showed that a certain minimum permeability is required in order to ensure the formation of a traveling thermal wave. It was found that by studying the non-dimensional governing parameters of the system one can ascribe characteristic values for these parameters for given initial and boundary conditions. This allows one to roughly predict the performance of a particular method on a particular reservoir given approximate values for initial and boundary conditions. Channelling and flow blockage due to carbon residue buildup impeded each method's performance. Blockage was found to be a result of imbalanced heating. Copyright 2012, Society of Petroleum Engineers.

Europium-directed self-assembly of a luminescent supramolecular gel from a tripodal terpyridine-based ligand.

Eu(III), the last piece in the puzzle: Europium-induced self-assembly of ligands having a C(3)-symmetrical benzene-1,3,5-tricarboxamide core results in the formation of luminescent gels. Supramolecular polymers are formed through hydrogen bonding between the ligands. The polymers are then brought together into the gel assembly through the coordination of terpyridine ends by Eu(III) ions (blue dashed arrow: distance between two ligands in the strand direction).

Understanding the kink formation in GaAs/InAs heterostructural nanowires

The kinks formation in heterostructural nanowires was observed to be dominant when InAs nanowires were grown on GaAs nanowires. Nanowires were grown through vapor-liquid-solid (VLS) mechanism in an MOCVD (metalorganic chemical vapor deposition) reactor. GaAs nanowires were grown in [1 1 1 ]B direction on a GaAs (1 1 1 )B substrate. When InAs nanowires grown on the GaAs nanowires, most of the InAs nanowires changed their growth directions from [1 1 1 ]B to other 〈111〉B directions. The kinks formation is ascribed to the large compressive misfit strain at the GaAs/InAs interface (7.2% lattice mismatch between GaAs and InAs) and the high mobility of indium species during MOCVD growth. The in-depth analysis of the kinks formation was done by growing InAs for short times on the GaAs nanowires and characterizing the samples. The hindrance to compressively strain InAs to form coherent layers with GaAs pushed the InAs/Au interfaces to the sides of the GaAs nanowires growth ends. New InAs/Au interfaces have generated at the sides of GaAs nanowires, due to lateral growth of InAs on GaAs nanowires. These new interfaces led the InAs nanowires growth in other 〈111〉B directions. The morphological and structural features of these heterostructural kinked nanowires were characterized using scanning electron microscopy (SEM) and transmission electron microscopy (TEM) techniques. © 2006 IEEE.

Dynamic compression of foam supported plates impacted by high velocity soil

The response of back-supported buffer plates comprising a solid face sheet and foam core backing impacted by a column of high velocity particles (sand slug) is investigated via a lumped parameter model and coupled discrete/continuum simulations. The buffer plate is either resting on (unattached) or attached to a rigid stationary foundation. The lumped parameter model is used to construct maps of the regimes of behaviour with axes of the ratio of the height of the sand slug to core thickness and the normalised core strength. Four regimes of behaviour are identified based on whether the core compression ends prior to the densification of the sand slug or vice versa. Coupled discrete/continuum simulations are also reported and compared with the lumped parameter model. While the model predicted regimes of behaviour are in excellent agreement with numerical simulations, the lumped parameter model is unable to predict the momentum transmitted to the supports as it neglects the role of elasticity in both the buffer plate and the sand slug. The numerical calculations show that the momentum transfer is minimised for intermediate values of the core strength when the so-called "soft-catch" mechanism is in play. In this regime the bounce-back of the sand slug is minimised which reduces the momentum transfer. However, in this regime, the impulse reduction is small (less than 10% of that transferred to a rigid structure). For high values of the core strength, the response of the buffer plate resembles a rigid plate with nearly no impulse mitigation while at low values of core strength, a slap event occurs when the face sheet impinges against the foundation due to full densification of the foam core. This slap event results in a significant enhancement of the momentum transfer to the foundation. The results demonstrate that appropriately designed buffer plates have potential as impulse mitigators in landmine loading situations. © 2013 Elsevier Ltd. All rights reserved.

Immunoglobulin joining (J) chain and its expression in the Chinese soft-shelled turtle (Pelodiscus sinensis)

The immunoglobulin (Ig) joining (J) chain plays an important role in the formation of polymeric Igs and their transport into secretions. In the present study, the cDNA sequence of J chain has been cloned from the Chinese soft-shelled turtle (Pelodiscus sinensis) by reverse transcription (RT)-PCR and rapid amplification of cDNA ends (RACE). The cDNA sequence is 2347 bp in length and contains an open reading frame of 480 bp encoding 160 aa including the signal sequence. The deduced amino acid sequence has a high degree of homology with that of an already reported turtle J chain (80.7%), and of chicken (71.3%). By using real-time quantitative RT-PCR analysis, a significant up-regulation of J-chain transcripts was observed in spleen, kidney and blood of turtles injected with inactivated Aeromonas hydrophila, indicating the immune role of J chain in response to bacterial infection. (C) 2009 Elsevier B.V. All rights reserved.

Antimicrobial activity-specific to Gram-negative bacteria and immune modulation-mediated NF-kappa B and Sp1 of a medaka beta-defensin

Defensins are a group of cationic antimicrobial peptides which play an important role in the innate immune system by exerting their antimicrobial activity against pathogens. In this study, we cloned a novel beta-defensin cDNA from medaka (Oryzias latipes) by rapid amplification of cDNA ends (RACE) technique. The full-length cDNA consists of 480 bp, and the open reading frame (CRF) of 189 bp encodes a polypeptide of 63 amino acids (aa) with a predicted molecular weight of 7.44 kDa. Its genomic organization was analyzed, and Southern blot detection confirmed that only one copy of beta-defensin exists in the medaka HNI strain. RT-PCR, Western blot and immunohistochemistry detections showed that the beta-defensin transcript and protein could be detected in eyes, liver, kidney, blood, spleen and gill, and obviously prevalent expression was found in eyes. Antimicrobial activity of the medaka beta-defensin was evaluated, and the antibacterial activity-specific to Gram-negative bacteria was revealed. Furthermore, the lipopolysaccharide (LPS), a major component of the outer membrane of Gram-negative bacteria, was demonstrated to be able to induce about 13-fol up-regulation of the beta-defensin within first 12 h. In addition, promoter and promoter mutagenesis analysis were performed in the medaka beta-defensin. A proximal 100 base pair(bp) sequence (+26 to -73)and the next 1700 bp sequence (-73 to -1755) were demonstrated to be responsible for the basal promoter activity and for the transcription regulation. Three nuclear factor kappa B (NF-kappa B) cis-elements and a Sp1 cis-element were revealed by mutagenesis analysis to exist in the 5' flanking sequence, and they were confirmed to be responsible for the up-regulation of medaka beta-defensin stimulated by LPS. And, the Sp1 cis-element was further revealed to be related to the basal promoter activity, and transcriptional factor II D (TFIID) was found to be in charge of the gene transcription initiation. All the obtained data suggested that the novel medaka beta-defensin should have antimicrobial activity-specific to Gram-negative bacteria, and the antibacterial immune function should be modulated by NF-kappa B and Sp1. (C) 2008 Elsevier Ltd. All rights reserved.

Metallothionein-2 gene from the mandarin fish Siniperca chuatsi: cDNA cloning, tissue expression, and immunohistochemical localization

The metallothionein-2 (MT-2) gene was isolated from the mandarin fish, one of the most important industrial aquatic animals in China, by using rapid amplification of cDNA ends (RACE). The deduced amino acid sequence of MT-2 comprised 60 amino acids and showed approximately 62.3% identity to human metallothionein. Its promoter region was amplified by thermal asymmetric interlaced polymerase chain reaction (TAIL-PCR). The MT-2 gene consists of 3 exons and 2 introns, extending approximately 900 bp of genomic sequence. Phylogenetic analysis clearly demonstrated that MT-2 formed a clade with fish metallothionein. The promoter region contained 5 putative metal-regulatory elements (MREs) and 1 TATA box. Real-time quantitative RT-PCR analysis revealed that MT-2 transcripts were significantly increased in the brain and gills and were stable in the muscles, liver, and trunk kidney in Cd2+-stimulated fish. Western blotting analysis demonstrated that the protein of the MT-2 gene was expressed mainly in the gills, liver, heart, trunk kidney, muscle, and intestine; it was weakly detected in the brain and head kidney. Moreover, the MT-2 protein was immunohistochemically detected in the cytoplasm in the liver and trunk kidney. All the above results revealed that the mandarin fish MT-2 would be a useful biomarker for metal pollution. (C) 2008 Published by Elsevier Inc.

Cloning, characterization and expression analysis of SIMP (source of immunodominant MHC-associated peptides) in grass carp Ctenopharyngodon idella

SIMP (source of immunodominant MHC-associated peptides) plays a key rote in N-linked glycosylation with the active site of oligosaccharyltransferase, being the source of MHC-peptides in the MHC I presentation pathway. In the present study, the SIMP gene has been cloned from grass carp Ctenopharyngodon idella by rapid amplification of cDNA ends (RACE). The full length of the cDNA sequence is 4384 bp, including a 1117 bp 5' UTR (untranslated region), a 2418 bp open reading frame, and a 849 bp 3' UTR. The deduced amino acids of the grass carp SIMP (gcSIMP) are a highly conserved protein with a STT3 domain and 11 transmembrane regions. The gcSIMP spans over more than 24,212 bp in length, containing 16 exons and 15 introns. Most encoding exons, except the first and the 15th, have the same length as those in human and mouse. The gcSIMP promoter contains many putative transcription factor binding sites, such as Oct-1, GCN4, YY1, Sp1, Palpha, TBP, GATA-1, C/EBP beta, and five C/EBP alpha binding sites. The mRNA expression of gcSIMP in different organs was examined by real-time PCR. The gcSIMP was distributed in all the organs examined, with the highest level in brain, followed by the level in the heart, liver, gill, trunk kidney, muscle, head kidney, thymus, and the lowest level in spleen. Furthermore, the recombinant gcSIMP has been constructed successfully and expressed in Escherichia coli by using pQE-40 vector, and the polyclonal antibody for rabbit has been successfully obtained, which was verified to be specific. Identification of gcSIMP will help to explore the function in fish innate immunity. (c) 2007 Elsevier Ltd. All rights reserved.

Seasonal patterns of sedimentation and their associations with benthic communities in Xiangxi Bay of the Three Gorges Reservoir, China

Sedimentation variables and benthic community data were collected at seven stations during four seasons in Xiangxi Bay of the Three Gorges Reservoir, China. Summer, the season of highest discharge into the reservoir, was characterized by the extreme sediment loading. The benthic macroinvertebrate community was dominated by oligochaetes across all seasons at most stations. In winter/spring, macroinvertebrate density and richness increased. Correspondence analysis showed that community structure differed among stations at the two ends of the bay in winter and among almost all stations in spring, However, no variable associated with sedimentation appeared to be associated with differences in the community.

Cloning and characterization of interferon stimulated genes Viperin and ISG15, and their promoters from snakehead Channa argus

By suppression subtractive hybridization, rapid amplification of cDNA ends and gene walking methods, interferon stimulated genes (ISGs), Viperin and ISG15, and their promoters have been cloned and characterized from snakehead Channa argus. The Viperin cDNA was found to be 1474 nt and contain an open reading frame (ORF) of 1059 nt that translates into a putative peptide of 352 amino acid (aa). The putative peptide of Viperin shows high identity to that in teleosts and mammals except for the N-terminal 70 aa. The ISG15 cDNA was found to be 758 nt and contain an ORF of 468 nt that translates into a putative peptide of 155 aa. The putative peptide of ISG15 is composed of two tandem repeats of ubiquitin-like (UBL) domains, and a canonical conjugation motif (LRGG) at C-terminal. Viperin and ISG15 promoter regions were characterized by the presence of interferon stimulating response elements (ISRE) and gamma-IFN activation sites (GAS). ISRE is a feature of IFN-induced gene promoter and partially overlaps interferon regulatory factor (IRF) 1 and IRF2 recognition sites. GAS is responsible for the gamma-IFN mediated transcription. One conserved site for NF-kappa B was found in the promoter region of Viperin. This is the first report of conservative binding motif for NF-kappa B in accordance with the consensus sequence (GGGRN-NYYCC) among teleost ISG promoters. Moreover, there were also TATA, CAAT and Sp1 transcription factor sites in Viperin and ISG15 promoters. In 5' untranslated region (UTR), snakehead ISG15 gene contains a single intron, which differs from Viperin gene. The transcripts of Vipeirn and ISG15 mRNA were mainly expressed in head kidney, posterior kidney, spleen and gill. The expression levels in liver were found to increase obviously in response to induction by IFN-inducer poly I : C.