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.

Dynamic centrifuge tests on shallow tunnel models in dry sand

In the framework of the Italian research project ReLUIS-DPC, a set of centrifuge tests were carried out at the Schofield Centre in Cambridge (UK) to investigate the seismic behaviour of tunnels. Four samples of dry sand were prepared at different densities, in which a small scale model of circular tunnel was inserted, instrumented with gauges measuring hoop and bending strains. Arrays of accelerometers in the soil and on the box allowed the amplification of ground motion to be evaluated; LVDTs measured the soil surface settlement. This paper describes the main results of this research, showing among others the evolution of the internal forces during the model earthquakes at significant locations along the tunnel lining. © 2010 Taylor & Francis Group, London.

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

Shock transmission in a coupled beam system

This paper investigates the circumstances under which high peak acceleration can occur in the internal parts of a system when subjected to impulsive driving on the outside. Motivating examples include the design of packaging for transportation of fragile items. The system is modelled in an idealised form using two beams coupled with point connections. A Rayleigh-Ritz model of such coupled beams was validated against measurements on a particular beam system, then the model was used to explore the acceleration response to impulsive driving in the time, frequency and spatial domains. This study is restricted to linear vibration response and additional mechanisms for high internal acceleration due to nonlinear effects such as internal impacts are not considered. Using Monte Carlo simulation in which the indirectly driven beam was perturbed by randomly placed point masses a wide range of system behaviour was explored. This facilitates identification of vulnerable configurations that can lead to high internal acceleration. The results from the study indicate the possibility of curve veering influencing the peak acceleration amplification. The possibility of veering within an ensemble was found to be dependent on the relative coupling strength of the modes. Understanding of the mechanism may help to avoid vulnerable cases, either by design or by preparatory vibration testing. © 2013 Elsevier Ltd.

Influence of tire geometry on the horn effect

A reciprocal-configuration Boundary Element Method calculation of acoustic radiation characteristics has been implemented for a generic tire geometry. The influence of the geometric parameters on the radiation characteristics has been studied. The degree of amplification of noise sources on the tire belt is strongly affected by the overall tire width. In contrast, the tire radius predominantly influences the pattern of the varying amplification around the belt, rather than its absolute level. Radiusing the tire's 'shoulder' region is potentially beneficial in terms of lowering amplification levels, for a tire of fixed overall width. However, it is less effective than maintaining sharp shoulders and reducing the overall width. Thus, for an acoustically optimal belted tire, the overall width should be as small as possible, even if this leads to a larger diameter. The width should not be increased in order to accommodate a radiused crown region. Copyright © (2012) by the Institute of Noise Control Engineering (INCE).

Superfluorescence in semiconductor lasers

An analysis is made of the conditions for the generation of superfluorescence pulses in an inverted medium of electron-hole pairs in a semiconductor. It is shown that strong optical amplification in laser semiconductor amplifiers characterised by αL ≫ 1 leads to suppression of phase re-laxation of the medium during the initial stages of evolution of superfluorescence and to formation of a macroscopic dipole from electron-hole pairs. Cooperative emission of radiation in this system results in generation of a powerful ultrashort pulse of the optical gain, which interacts coherently with the semiconductor medium. It is shown that coherent pulsations of the optical field, observed earlier by the author in Q-switched semiconductor lasers, are the result of superfluorescence and of the coherent interaction between the optical field and the medium.

Superfluorescence in semiconductor lasers

An analysis is made of the conditions for the generation of superfluorescence pulses in an inverted medium of electron-hole pairs in a semiconductor. It is shown that strong optical amplification in laser semiconductor amplifiers characterised by αL ≫ 1 (α is the small-signal gain and L is the amplifier length) leads to suppression of phase relaxation of the medium during the initial stages of evolution of superfluorescence and to formation of a macroscopic dipole from electron - hole pairs. Cooperative emission of radiation in this system results in generation of a powerful ultrashort pulse of the optical gain, which interacts coherently with the semiconductor medium. It is shown that coherent pulsations of the optical field, observed earlier by the author in Q-switched semiconductor lasers, are the result of superfluorescence and of the coherent interaction between the optical field and the medium.

2.1 GHz MOS mixer linearisation approach for IIP2 enhancement using multiple factor compensation

An integrated downconversion CMOS mixer incorporating a comprehensive compensation scheme is presented which aims to minimise second-order intermodulation distortion (IMD2). Unlike previously reported IMD2 calibration schemes which tune only one nonlinear factor at a time, the presented solution allows simultaneous adjustment of several different factors thus achieving a better compensation. The mixer has been implemented in UMC 0.18 μm CMOS to verify the proposed scheme and for comparison with alternative compensation methods. Measurements show that the solution described can improve the input intercept point (IIP2) by over 20 dB while maintaining good amplification and noise performance. IMD2 calibration results are presented and show useful advantages over other approaches. To the best of the authors' knowledge, this scheme for IMD2 calibration has not been previously reported. © The Institution of Engineering and Technology 2013.

Implementation of trailer steering control on a multi-unit vehicle at high speeds

A high-speed path-following controller for long combination vehicles (LCVs) was designed and implemented on a test vehicle consisting of a rigid truck towing a dolly and a semitrailer. The vehicle was driven through a 3.5 m wide lane change maneuver at 80 km/h. The axles of the dolly and trailer were steered actively by electrically-controlled hydraulic actuators. Substantial performance benefits were recorded compared with the unsteered vehicle. For the best controller weightings, performance improvements relative to unsteered case were: lateral tracking error 75% reduction, rearward amplification (RA) of lateral acceleration 18% reduction, and RA of yaw rate 37% reduction. This represents a substantial improvement in stability margins. The system was found to work well in conjunction with the braking-based stability control system of the towing vehicle with no negative interaction effects being observed. In all cases, the stability control system and the steering system improved the yaw stability of the combination. © 2014 by ASME.

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.

Multi-frequency operation of a mems vibration energy harvester by accessing five orders of parametric resonance

The mechanical amplification effect of parametric resonance has the potential to outperform direct resonance by over an order of magnitude in terms of power output. However, the excitation must first overcome the damping-dependent initiation threshold amplitude prior to accessing this more profitable region. In addition to activating the principal (1st order) parametric resonance at twice the natural frequency ω0, higher orders of parametric resonance may be accessed when the excitation frequency is in the vicinity of 2ω0/n for integer n. Together with the passive design approaches previously developed to reduce the initiation threshold to access the principal parametric resonance, vacuum packaging (< 10 torr) is employed to further reduce the threshold and unveil the higher orders. A vacuum packaged MEMS electrostatic harvester (0.278 mm3) exhibited 4 and 5 parametric resonance peaks at room pressure and vacuum respectively when scanned up to 10 g. At 5.1 ms-2, a peak power output of 20.8 nW and 166 nW is recorded for direct and principal parametric resonance respectively at atmospheric pressure; while a peak power output of 60.9 nW and 324 nW is observed for the respective resonant peaks in vacuum. Additionally, unlike direct resonance, the operational frequency bandwidth of parametric resonance broadens with lower damping. © Published under licence by IOP Publishing Ltd.

Bead-probe complex capture a couple of SINE and LINE family from genomes of two closely related species of East Asian cyprinid directly using magnetic separation

Background: Short and long interspersed elements (SINEs and LINEs, respectively), two types of retroposons, are active in shaping the architecture of genomes and powerful tools for studies of phylogeny and population biology. Here we developed special protocol to apply biotin-streptavidin bead system into isolation of interspersed repeated sequences rapidly and efficiently, in which SINEs and LINEs were captured directly from digested genomic DNA by hybridization to bead-probe complex in solution instead of traditional strategy including genomic library construction and screening. Results: A new couple of SINEs and LINEs that shared an almost identical 3'tail was isolated and characterized in silver carp and bighead carp of two closely related species. These SINEs (34 members), designated HAmo SINE family, were little divergent in sequence and flanked by obvious TSD indicated that HAmo SINE was very young family. The copy numbers of this family was estimated to 2 x 10(5) and 1.7 x 10(5) per haploid genome by Real-Time qPCR, respectively. The LINEs, identified as the homologs of LINE2 in other fishes, had a conserved primary sequence and secondary structures of the 3'tail region that was almost identical to that of HAmo SINE. These evidences suggest that HAmo SINEs are active and amplified recently utilizing the enzymatic machinery for retroposition of HAmoL2 through the recognition of higher-order structures of the conserved 42-tail region. We analyzed the possible structures of HAmo SINE that lead to successful amplification in genome and then deduced that HAmo SINE, SmaI SINE and FokI SINE that were similar in sequence each other, were probably generated independently and created by LINE family within the same lineage of a LINE phylogeny in the genomes of different hosts. Conclusion: The presented results show the advantage of the novel method for retroposons isolation and a pair of young SINE family and its partner LINE family in two carp fishes, which strengthened the hypotheses containing the slippage model for initiation of reverse transcription, retropositional parasitism of SINEs on LINEs, the formation of the stem loop structure in 3'tail region of some SINEs and LINEs and the mechanism of template switching in generating new SINE family.

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.

Genetic variation of fish parasite populations in historically connected habitats: undetected habitat fragmentation effect on populations of the nematode Procamallanus fulvidraconis in the catfish Pelteobagrus fulvidraco

Habitat fragmentation may have some significant effects on population genetic structure because geographic distance and physical barriers may impede gene flow between populations. In this study, we investigated whether recent habitat fragmentation affected genetic structure and diversity of populations of the nematode Procamallanus fulvidraconis in the yellowhead catfish, Pelteobagrus fin't4draco. The nematode was collected from 12 localities in 7 floodplain lakes of the Yangtze River. Using I I intersirnple sequence repeat markers, analysis of molecular variance showed that genetic diversity occurred mainly within populations (70.26%). Expected heterozygosity (He) of P. fulvidraconis was barely different between connected (0.2105) and unconnected lakes (0.2083). Population subdivision (Fst) between connected lakes (0.2177) was higher than in unconnected lakes (0. 1676). However, the connected and unconnected lakes did not Cluster into 2 clades. A Mantel test revealed significant positive correlation between genetic and geographic distances (R = 0.5335, P < 0.01). These results suggest that habitat fragmentation did not cause genetic differentiation among populations or a reduction of diversity in isolated populations of P. fulvidraconis. At least 2 factors may increase the dispersal range of the nematode, i.e., flash flooding in summer and other species of fish that may serve as the definitive hosts. Moreover, lake fragmentation is probably a recent process; population size of the nematode in these lakes is large enough to maintain Population structure.