A transglutaminase from Chinese shrimp (Fenneropenaeus chinensis), full-length cDNA cloning, tissue localization and expression profile after challenge

Transglutaminase can catalyze the cross-linking reaction between soluble clotting protein molecules from the plasma for prevention of excess blood loss from a wound and obstructing micro-organisms from invading the wound in crustaceans. A novel transglutaminase (FcTG) gene was cloned from hemocytes of Chinese shrimp Fenneropenaeus chinensis by 3' and 5' rapid amplification of cDNA ends (RACE) PCR. The full-length cDNA consists of 2972 bp, encoding 757 amino acids with a calculated molecular mass of 84.96 kDa and a theoretical isoelectric point of 5.61. FcTG contains a typical transglutaminase-like homologue (TGc domain: E-value = 1.94e-38). Three catalytic sites (Cys-324, His-391 and Asp-414) are present in this domain. The deduced amino acid sequence of FcTG showed high identity with black tiger shrimp TG, kuruma shrimp TG and crayfish TG. Transcripts of FcTG mRNA were mainly detected in gill, lymphoid organ and hemocytes by RT-PCR. RNA in situ hybridization further confirmed that FcTG was constitutively expressed in hemocytes both in the circulatory system and lymphoid organ. The variation of mRNA transcription level in hemocytes and lymphoid organ following injection of killed bacteria or infection with white spot syndrome virus (WSSV) was quantified by RT-PCR. The up-regulated expression of FcTG in shrimp lymphoid organ following injection of bacteria indicates that it is inducible and might be associated with bacterial challenge. (c) 2006 Elsevier Ltd. All rights reserved.

Dissection and localization of the immunostimulating domain of Edwardsiella tarda FliC

Bacterial flagellin is known to induce potent immune response in vertebrate systems via the toll-like receptor (TLR) 5. As a result, flagellin has been studied extensively as a vaccine adjuvant. In a previous study, we examined the vaccine and adjuvant potentials of the flagellin (FliC) of the fish pathogen Edwardsiella tarda. We found that E. tarda FliC induced low protective immunity by itself but could function as a molecular adjuvant and potentiate the specific immune response induced by the E. tarda antigen Eta6. Since FliC is a large protein and organized into distinct structural domains, we wondered whether the immunostimulating effect observed with the full-length protein could be localized to a certain region. To investigate this question, we in the present study dissected the FliC protein into several segments according to its structural features: (i) N163, which consists of the conserved N-terminal 163 residues of FliC; (ii) M160, which consists of the variable middle 160 residues; (iii) C94, which consists of the conserved C-terminal 94 residues; (iv) NC257, which is an artificial fusion of N163 and C94. To examine the adjuvanticity of the FliC fragments, DNA vaccine plasmids expressing FliC fragments in fusion with Eta6 were constructed and used to immunize Japanese flounder. The results showed that N163 produced the best adjuvant effect, which, in respect to improvement in the relative percent survival of the vaccinated fish, was comparable to that of the full-length FliC. None of the other FliC fragments exhibited apparent immunopotentiating effect. Further analysis showed that N163 enhanced the production of serum specific antibodies and, like full-length FliC, significantly upregulated the expression of the genes that are possibly involved in innate and adaptive immunity. These results indicate that N163 is the immunodominant region of FliC and suggest that E. tarda FliC may induce immune responses in Japanese flounder via mechanisms alternative to that involving TLR5. (C) 2010 Elsevier Ltd. All rights reserved.

Expression and localization of a novel phosducin-like protein from amphioxus Branchiostoma belcheri

A full length amphioxus cDNA, encoding a novel phosducin-like protein (Amphi-PhLP), was identified for the first time from the gut cDNA library of Branchiostoma belcheri. It is comprised of 1 550 bp and an open reading frame (ORF) of 241 amino acids, with a predicted molecular mass of approximately 28 kDa. In situ hybridization histochemistry revealed a tissue-specific expression pattern of Amphi-PhLP with the high levels in the ovary, and at a lower level in the hind gut and testis, hepatic caecum, gill, endostyle, and epipharyngeal groove, while it was absent in the muscle, neural tube and notochord. In the Chinese Hamster Ovary (CHO) cells transfected with the expression plasmid pEGFP-N1/Amphi-PhLP, the fusion protein was targeted in the cytoplasm of CHO cells, suggesting that Amphi-PhLP is a cytosolic protein. This work may provide a framework for further understanding of the physiological function of Amphi-PhLP in B. belcheri.

Identification, expression, function and localization of a DUF985 domain-containing hypothetical gene from amphioxus Branchiostoma belcheri

The progress in genome sequencing has led to an increasing submission of uncharacterized hypothetical genes with the domain of unknown function, DUF985, in GenBank, and none of these genes is related to a known protein. We therefore underwent an experimental study to identify the function of a DUF985 domain-containing hypothetical gene BbDUF985 (GenBank Accession No. AY273818) isolated from amphioxus Branchiostoma belcheri (B. belcheri). BbDUF985 was successfully expressed in both prokaryotic and eukaryotic systems, and its recombinant proteins expressed in both systems definitely exhibited an activity of phosphoglucose isomerase (PGI). Both tissue-section in situ hybridization and immunohistochemistry demonstrated that BbDUF985 was expressed in a tissue-specific manner, with most abundant levels in the hepatic caecum and ovary. In CHO cells transfected with the expression plasmid pEGFP-N1/BbDUF985, the fusion protein was targeted in the cytoplasm of CHO cells, suggesting that BbDUF985 is a cytosolic protein. In contrast, Western blotting indicated that BbDUF985 was also present in amphioxus humoral fluids, suggesting that it exists as a secreted protein as well. Our study provided a framework for further understanding the biochemical properties and physiological function of DUF985-containing hypothetical proteins in other species. (c) 2008 Elsevier Inc. All rights reserved.

Localization of the electric-field distribution in graded core-shell metamaterials

The local electric-field distribution has been investigated in a core-shell cylindrical metamaterial structure under the illumination of a uniform incident optical field. The structure consists of a homogeneous dielectric core, a shell of graded metal-dielectric metamaterial, embedded in a uniform matrix. In the quasistatic limit, the permittivity of the metamaterial is given by the graded Drude model. The local electric potentials and hence the electric fields have been derived exactly and analytically in terms of hypergeometric functions. Our results showed that the peak of the electric field inside the cylindrical shell can be confined in a desired position by varying the frequency of the optical field and the parameters of the graded profiles. Thus, by fabricating graded metamaterials, it is possible to control electric-field distribution spatially. We offer an intuitive explanation for the gradation-controlled electric-field distribution.

用于移动机器人自定位的视觉路标设计

为了实现室内移动机器人的自定位,提出了一种简易美观的新型视觉人工路标以及基于对数极坐标系投影直方图的路标识别方法,并用基于共面四点的位姿估计算法计算机器人位姿。实验结果说明,路标检测具有很高的鲁棒性;路标识别方法抗噪声和形变的能力强;位姿精度足够满足室内移动机器人自定位的需要。

基于视觉的室内移动机器人自定位技术研究

在关于移动机器人的诸多研究领域中，机器人自定位是十分关键的技术，是实现机器人自主运动和其他任务的基础，而且涉及领域广泛，有很多难点有待解决，因而是一个具有重要研究价值的课题。 本论文以沈阳新松机器人股份有限公司自主研发的家庭服务机器人为研发平台，系统地研究了基于计算机视觉的室内移动机器人自定位问题，成功设计了基于单目视觉人工路标以及粒子滤波的室内移动机器人自定位系统。 本文首先根据室内移动机器人自主导航定位的要求，设计了一种简易美观的新型视觉人工路标，并且研究与实现了该路标的实时准确检测以及不同路标的识别。 其次，在位姿计算方面，本文研究了共面P4P（4点透视）问题的解法及其在位姿计算方面的应用，并分析比较了两种不同P4P解法的优缺点，成功地将两种算法结合起来用于机器人位姿计算。 最后，在机器人自定位方面，本文将单目彩色摄像机作为传感器，在基于贝叶斯滤波理论的自定位理论框架下，利用粒子滤波自定位方法融合视觉信息与码盘信息，实现了自主移动机器人的自定位。 实践证明，本文设计的基于单目视觉人工路标的自定位系统能够成功地应用在室内移动机器人上，具有较高的应用推广价值。

基于局部子图匹配的SLAM方法

针对现有的SLAM解决方法在机器人被"绑架"时失效的问题,提出了基于局部子图匹配的方法。该方法对现有的SLAM解决构架进行了改进,提出交点最优匹配的特征相关算法,并且将奇异值分解方法引入机器人定位。最后,在结构化环境下将本方法和基于扩展卡尔曼滤波器的方法进行比较,讨论了基于局部子图匹配的方法在结构化环境中解决机器人"绑架"问题的有效性和可行性。

一种输电线路巡检机器人控制系统的设计与实现

介绍了一种超高压输电线路巡检机器人控制系统的设计与实现方法.根据巡检作业任务的要求,采用遥控与局部自主相结合的控制模式实现巡检机器人沿线行走及跨越障碍.设计了巡检机器人有限状态机模型,实现了机器人遥控与局部自主控制的有机结合.采用基于激光传感器定位的方法实现了巡检机器人的自主越障控制.实验结果表明,该机器人可实现沿线行走及自主跨越障碍,从而验证了控制系统设计的有效性与合理性.

基于视觉伺服的输电线巡检机器人抓线控制

巡检机器人在越障时,需要完成机器人手臂的准确抓线控制.结合输电线的几何特征和摄像机成像原理,提出了一种基于单摄像机的立体视觉方法来确定输电线的位置和姿态.基于该定位方法及视觉伺服理论,建立机械手抓线伺服控制模型.利用自行研制的巡检机器人进行了视觉伺服抓线实验;实验结果验证了该方法的有效性.

基于立体视觉的虚拟机械臂平面定位研究

结合星球探测的应用背景,对漫游车的工作方式进行了研究,针对车载机械臂开发了一套基于立体视觉的机械臂平面定位仿真系统。该系统依靠虚拟现实技术,通过虚拟机械臂对三维重建得到的平整物体表面的定位仿真得到机械臂的各关节参数,以此指导真实机械臂的运动。论述了基于立体视觉的机械臂定位机理和基于OpenGL的虚拟机械臂的实现过程。采用VC++构建了仿真实验平台,进行了定位实验,获得了较高的定位精度。

超高压输电线路巡检机器人两种越障定位方法比较

将两种基于传感器信息的导线定位方法的特点和差异进行了分析和对比,得出方法Ⅱ的解算能力强于方法Ⅰ的解算能力,但由于方法Ⅱ的定位精度低于方法Ⅰ的定位精度,因此,选用方法Ⅰ进行野外作业。

输电线路巡检机器人越障控制研究

介绍了超高压输电线路巡检机器人越障控制方法。根据巡检作业任务的要求,采用遥控与局部自主控制相结合的方法,实现了巡检机器人沿线行走及跨越障碍的功能。采用基于单目摄像头定位和视觉伺服的方法,实现了巡检机器人的自主越障控制。实验结果表明,该机器人可沿线行走并自主跨越障碍,从而验证了控制系统设计的有效性与合理性。

自主移动机器人足球比赛视觉定位方法综述

综述了RoboCup足球赛中全自主移动机器人基于视觉的定位技术,包括机器人自定位和多机器人协作物体定位.介绍了定位技术的发展情况与分类.从机器人环境构建形式的不同以及先验位姿和概率方法的应用与否等方面,系统地分析和比较了各种自定位方法.对于多机器人协作物体定位,阐述了静态方法和动态跟踪方法.总结了定位过程中需要重点研究的传感器模型构建、图像处理、特征匹配以及协作过程涉及的相关问题.最后就视觉定位存在的问题和技术发展趋势进行了讨论.

移动机器人几何-拓扑混合地图的构建及自定位研究

基于墙角、房门和通路等高级环境特征的辨识与提取,依据几何和拓扑环境模型完成混合地图的构建,并根据混合地图的特点,提出在局部几何环境采用扩展卡尔曼滤波算法实现移动机器人的位姿跟踪,而在拓扑地图的节点位置则根据绑定的高级环境特征进行位姿再校正的混合定位方法.将该方法应用于实际移动机器人平台,所得结果证明了方法的有效性和实用性.