Contrast features of CpG islands in the promoter and other regions in the dog genome

The recent release of the domestic dog genome provides us with an ideal opportunity to investigate dog-specific genomic features. In this study, we performed a systematic analysis of CpG islands (CGIs), which are often considered gene markers, in the dog

CpG islands or CpG clusters: how to identify functional GC-rich regions in a genome?

Background: CpG islands (CGIs), clusters of CpG dinucleotides in GC-rich regions, are often located in the 5' end of genes and considered gene markers. Hackenberg et al. ( 2006) recently developed a new algorithm, CpGcluster, which uses a completely diffe

Comparative analysis of CpG islands in four fish genomes

There has been much interest in CpG islands (CGIs), clusters of CpG dinucleotides in GC-rich regions, because they are considered gene markers and involved in gene regulation. To date, there has been no genome-wide analysis of CGIs in the fish genome. We

Sequence context analysis in the mouse genome: Single nucleotide polymorphisms and CpG island sequences

A genome-wide view of sequence mutability in mice is still limited, although biologists usually assume the same scenario for mice as for humans. In this study, we examined the sequence context in the local environment of 482,528 mouse single nucleotide po

Protective immunity induced by CpG ODNs against white spot syndrome virus (WSSV) via intermediation of virus replication indirectly in Litopenaeus vannamei

The worldwide shrimp culture is beset with diseases mainly caused by white spot syndrome virus (WSSV) and suffered huge economic losses, which bring out an urgent need to develop the novel strategies to better protect shrimps against WSSV. In the present study, CpG-rich plasmid pUC57-CpG, plasmid pUC57 and PBS were employed to pretreat shrimps comparatively to evaluate the protective effects of CpG ODNs on shrimps against WSSV. The survival rates, WSSV copy numbers, and antiviral associated factors (Dicer, Argonaute, STAT and ROS) were detected in Litopenaeus vannamei. There were higher survival proportion, lower WSSV copy numbers, and higher mRNA expression of Dicer and STAT in pUC57-CpG-pretreatment shrimps than those in pUC57- and PBS-pretreatment shrimps after WSSV infection. The Argonaute mRNA expression in pUC57-CpG-, pUC57- and PBS-pretreatment shrimps after WSSV infection was significantly higher than that of shrimps post PBS stimulation on the first day. The ROS levels in pUC57-CpG-pretreatment shrimps post secondary stimulation of PBS were significantly higher than those post WSSV infection on the first day. These results together demonstrated that pUC57-CpG induced partial protective immunity in shrimps against WSSV via intermediation of virus replication indirectly and could be used as a potential candidate in the development of therapeutic agents for disease control of WSSV in L. vannamei. (C) 2009 Elsevier Ltd. All rights reserved.

The effects of CpG-C oligodeoxynucleotides on innate immune responses in Eriocheir sinensis (H. Milne-Edwards, 1853)

CpG oligodeoxynucleotides (ODNs) can stimulate the immune system, and therefore are widely used as a therapeutic vaccination and immune adjuvant in human. In the present study, CpG-C, a combination of A- and B-class ODN, was injected into Chinese mitten crab Eriocheir sinensis at three doses (0.1, 1 and 10 mu g crab-1), and the reactive oxygen species (ROS) levels, activities of total intracellular phenoloxidase (PO) and lysozyme-like activities, the mRNA transcripts of EsproPO, EsCrustin and EsALF were assayed to evaluate its modulating effects on the immune system of crab. The ROS levels in all treated and control groups were significantly increased from 6 to 24 h, except that ROS in 0.1 mu g CpG-C-treated crabs was comparable to that of the blank at 6 h. The PO activity was significantly enhanced and EsproPO transcripts were down-regulated (P < 0.01) at 6 h after the injection of 0.1 mu g CpG-C, with no significant changes in the other dosage treatments. The lysozyme-like activities and EsCrustin transcripts in the CpG-C-treatment groups were significantly higher than those of controls. The mRNA expression of EsALF remained almost constant in all the groups during the treatment. These results collectively suggested that CpG-C could activate the immune responses of E. sinensis, and might be used as a novel immunostimulant for disease control in crabs.

The effects of CpG-C oligodeoxynucleotides on innate immune responses in Eriocheir sinensis (H. Milne-Edwards, 1853)

CpG oligodeoxynucleotides (ODNs) can stimulate the immune system, and therefore are widely used as a therapeutic vaccination and immune adjuvant in human. In the present study, CpG-C, a combination of A- and B-class ODN, was injected into Chinese mitten crab Eriocheir sinensis at three doses (0.1, 1 and 10 mu g crab-1), and the reactive oxygen species (ROS) levels, activities of total intracellular phenoloxidase (PO) and lysozyme-like activities, the mRNA transcripts of EsproPO, EsCrustin and EsALF were assayed to evaluate its modulating effects on the immune system of crab. The ROS levels in all treated and control groups were significantly increased from 6 to 24 h, except that ROS in 0.1 mu g CpG-C-treated crabs was comparable to that of the blank at 6 h. The PO activity was significantly enhanced and EsproPO transcripts were down-regulated (P < 0.01) at 6 h after the injection of 0.1 mu g CpG-C, with no significant changes in the other dosage treatments. The lysozyme-like activities and EsCrustin transcripts in the CpG-C-treatment groups were significantly higher than those of controls. The mRNA expression of EsALF remained almost constant in all the groups during the treatment. These results collectively suggested that CpG-C could activate the immune responses of E. sinensis, and might be used as a novel immunostimulant for disease control in crabs.

Identification and analysis of the immune effects of CpG motifs that protect Japanese flounder (Paralichthys olivaceus) against bacterial infection

CpG-containing oligodeoxynucleotides (ODNs) are known to be immunostimulatory in vertebrate systems and can activate both innate and adaptive immune responses. In this report, we described the selection, identification, and analysis of CpG motifs with immunoprotective effects in Japanese flounder. Sixteen CpG ODNs were synthesized and examined for the ability to inhibit bacterial dissemination in Japanese flounder blood. Four ODNs with the strongest inhibitory effects were selected and mixed to form ODNs 4M. In addition, a plasmid, pCN6, was constructed that contains the sequences of the four selected ODNs. When administered into Japanese flounder via intraperitoneal injection, both ODNs 4M and pCN6 could, in dose and time dependent manners, afford short-term protection against the infections of two different bacterial pathogens. Immunological analyses showed that ODNs 4M and, especially, pCN6 activated head kidney macrophages and enhanced serum bactericidal activity via probably the alternative pathway of complement activation. When used as a DNA vaccine to immunize Japanese flounder, pCN6 conferred apparent protections (42.9% and 52.6%, respectively, in terms of relative percent survival) against the challenges of two different fish pathogens at 4-week post-vaccination. Transcriptional analysis showed that vaccination with pCN6 upregulated the expression of the genes encoding NKEF, MHC II alpha, IL-1 beta, Mx, and MHC I alpha. These results demonstrate that ODNs 4M and pCN6 are immunostimulatory in Japanese flounder and can induce short- and long-term nonspecific protections against bacterial infections. (C) 2010 Elsevier Ltd. All rights reserved.

Identification and analysis of a CpG motif that protects turbot (Scophthalmus maximus) against bacterial challenge and enhances vaccine-induced specific immunity

Oligodeoxynucleotides (ODNs) containing unmethylated CpG motifs in certain contexts are known to be immunostimulatory in vertebrate systems. CpG ODNs with immune effects have been identified for many fish species but, to our knowledge, not for turbot. In this study, a turbot-effective CpG ODN, ODN 205, was identified and a plasmid, pCN5, was constructed which contains the CpG motif of ODN 205. When administered into turbot via intraperitoneal (i.p.) injection, both ODN 205 and pCN5 could (i) inhibit bacterial dissemination in blood in dose and time dependent manners, and (ii) protect against lethal bacterial challenge. Immunological analyses showed that in vitro treatment with ODN 205 stimulated peripheral blood leukocyte proliferation, while i.p. injection with ODN 205 enhanced the respiratory burst activity, chemiluminescence response, and acid phosphatase activity of turbot head kidney macrophages. pCN5 treatment-induced immune responses similar to those induced by ODN 205 treatment except that pCN5 could also enhance serum bactericidal activity in a calcium-independent manner. To examine whether ODN 205 and pCN5 had any effect on specific immunity, ODN 205 and pCN5 were co-administered into turbot with a Vibrio harveyi subunit vaccine, DegQ. The results showed that pCN5, but not ODN 205, significantly increased the immunoprotective efficacy of DegQ and enhanced the production of specific serum antibodies in the vaccinated fish. Further analysis indicated that vaccination with DegQ in the presence of pCN5 upregulated the expression of the genes encoding MHC class II alpha, IgM, Mx, and IL-8 receptor. Taken together, these results demonstrate that ODN 205 and pCN5 can stimulate the immune system of turbot and induce protection against bacterial challenge. In addition, pCN5 also possesses adjuvant property and can potentiate vaccine-induced specific immunity. (C) 2010 Elsevier Ltd. All rights reserved.

蛇形机器人步态转换CPG控制器

蛇具有细长无肢的身体、独特的半球型关节,使其可在神经系统控制下完成与环境相适应的多种节律运动.基于对该节律运动机制的分析,给出蛇运动神经系统的主要功能特性.首次应用双向循环抑制CPG模拟蛇节律运动发生机制,控制蛇形机器人组合关节,实现3种典型运动.通过单向激励串联该类CPG构成神经网络,给出该神经网络产生振荡输出的必要条件.用不同高级控制神经元命令激活下的输出,实现蛇形机器人典型运动模式的自动转换.通过动力学仿真和实验验证该CPG控制器产生不同节律运动模式的有效性.为蛇节律运动模式发生机制建模提供新方法.

基于循环抑制CPG模型控制的蛇形机器人三维运动

具有三维运动能力和独特的节律运动方式,使生物蛇能在复杂的地形环境中生存.大多数动物节律运动是由中央模式发生器(Centralpatterngenerator,CPG)控制的.以此为理论依据,首次以循环抑制建模机理构建蛇形机器人组合关节运动控制的CPG模型.证明该模型是节律输出型CPG中微分方程维数最少的.采用单向激励方式连接该类CPG构建蛇形机器人三维运动神经网络控制体系,给出该CPG网络产生振荡输出的必要条件.应用蛇形机器人动力学模型仿真得到控制三维运动的CPG神经网络参数,利用该CPG网络的输出使“勘查者”成功实现三维运动.该结果为建立未探明的生物蛇神经网络模型提供了一种全新的方法.

基于循环抑制CPG模型的蛇形机器人控制器

依据生物利用中央模式发生器(Central pattern generator,CPG)的自激行为产生有节律的协调运动适应多种环境,基于循环抑制CPG建模理论设计了蛇形机器人CPG控制器模型,分析了单个神经元、循环抑制CPG以及该控制器模型的稳定性,并把该控制器应用到一个结合蛇形机器人“勘查者-Ⅰ”动力学特性的仿真模型,得到了实现蜿蜒运动的CPG控制器参数,进而研究了调节S波个数、身体构形曲率、蜿蜒运动速度以及运动轨迹曲率的CPG控制器参数设定策略。此外,“勘查者-Ⅰ”应用该CPG控制器的输出成功实现了蜿蜒运动。该研究结果为设计人工CPG控制器提供了一个可行的方法。

基于循环抑制CPG模型控制的蛇形机器人蜿蜒运动

根据生物蛇和蛇形机器人的结构及运动特点,应用循环抑制CPG建模理论构建了蛇形机器人神经网络模型;利用蛇形机器人模型,仿真验证了CPG模型对蜿蜒运动控制的有效性;提出并验证了实现有目的转弯控制的CPG参数调节方法．最后,给出了今后的研究方向．

基于循环抑制CPG的蛇形机器人运动控制方法

蛇具有细长无肢的身体、独特的半球形关节，使其可在神经系统控制下完成与环境相适应的多种节律运动。模仿蛇的运动机理和行为方式而设计的蛇形机器人克服了轮腿式机器人的缺点，增加了机器人的运动方式，扩大了机器人的应用范围。但应用传统的控制策略实现蛇形机器人运动控制遇到了很难克服的问题。随着社会经济与科技的发展，研究人员把从蛇运动神经系统研究中得到的启示应用到蛇形机器人上，希望不仅可以解决其运动控制问题，更能在构型、步态及控制机制上皆可展示蛇的特征。 生物学家已经证明动物的节律运动是其低级神经中枢的自激行为，是由中枢模式发生器(Central Pattern Generator，CPG)控制的。中枢模式发生器是一种能够在缺乏有规律的感知和中枢控制输入的情况下，产生有节奏模式输出的神经网络。 本文以国家自然科学基金课题《基于CPG的蛇形机器人控制方法研究》和国家“863”高技术计划资助项目《具有环境适应能力的蛇形机器人的研究》为依托，突破以相互抑制机理研究CPG的传统观点，首次创新性地提出应用循环抑制(Cyclic Inhibition, CI)机理来研究蛇形机器人的CPG建模与实现问题。本研究涵概了神经元模型的特性分析、蛇形机器人关节循环抑制CPG建模理论、蛇形机器人循环抑制CPG神经网络稳定性分析以及典型步态的生成方法、循环抑制CPG神经网络控制蛇形机器人蜿蜒运动参数设定策略、应用动力学仿真和实验对该CPG控制方法有效性的验证。 首先，本文介绍了两个用于CPG建模研究的蛇形机器人“勘查者”和“勘查者-I”。给出各自机械系统、控制系统的构成和动力学仿真平台。 其次，详细分析了神经元以及传统的相互抑制(Mutual Inhibition, MI)CPG的特性。从工程角度首次创新性地应用循环抑制建模理论构建了蛇形机器人CPG模型，并对其稳定性进行了深入的分析。首次证明持续型神经元构成的单向循环抑制(Unilateral Cyclic Inhibition, UCI) CPG是能产生振荡输出CPG中微分方程数量最少的，而且其产生振荡输出的机理完全不同于传统的相互抑制CPG。其不需要具备调整功能，只需要神经元之间强的单向循环抑制连接。 第三，首次应用单向激励连接循环抑制CPG构成蛇形机器人神经网络系统。分析了其稳定性，给出其产生振荡输出的条件。通过仿真和实验验证了循环抑制CPG神经网络实现典型步态(蜿蜒运动、伸缩运动和侧向运动)的有效性。首次应用双向循环抑制(Bidirectional Cyclic Inhibition, BCI)CPG神经网络在不同高级控制神经元命令激活下的输出实现蛇形机器人典型运动步态之间的转换。为蛇节律运动生成机制建模提供了新方法。 最后，从实时性、控制方便性等工程应用的角度，对单向循环抑制CPG神经网络实现蛇形机器人蜿蜒运动控制进行了深入的分析。给出了S-波形、幅值、运动速度和运动轨迹曲率的参数设定策略。该系统应用首CPG自激励权重调解成功解决了传统CPG控制系统中CPG的个数比蛇形机器人关节数多一个的问题，并用其实现了一种独特的转弯控制策略。 综上，为蛇形机器人运动控制提供了全新的方法。

Imunização de bovinos com MSP1a e MSP2 recombinantes de Anaplasma marginale com CpG ODN 2006 como adjuvante, e desafio com isolado heterólogo.

A anaplasmose é uma importante enfermidade de bovinos de áreas tropicais e subtropicais do mundo, causada pela riquétsia intra-eritrocítica Anaplasma marginale. A vacinação tem sido a forma mais econômica e eficiente de controlar a anaplasmose bovina. Nos últimos anos, esses estudos têm se concentrado nas proteínas de membrana da riquétsia, sobretudo MSP1a e MSP2. Este trabalho teve como objetivos avaliar o grau de proteção induzido pelas proteínas de membrana MSP1a e MSP2 recombinantes de A. marginale, associadas com adjuvante CpG ODN 2006, perante desafio heterólogo e avaliar a resposta imune gerada. Novilhos da raça Aberdeen Angus foram imunizados três vezes com 200 ?g de MSP1a e/ou MSP2 recombinantes, associadas com CpG ODN 2006 e alúmen. Posteriormente, foram desafiados com 3 x 107 eritrócitos infectados com isolado heterólogo de A. marginale. Os animais experimentais apresentaram quadro clínico de anaplasmose (redução do volume globular, febre e riquetsemias detectáveis por distensões sangüíneas coradas). Não foram detectadas diferenças significativas entre os grupos imunizados e os controles quanto ao percentual de redução do volume globular, riquetsemias máximas e temperaturas retais máximas, indicando que as imunizações não foram protetoras. A despeito da significativa produção de IgG total contra MSP1a e MSP2, detectada no dia do desafio, os animais imunizados apresentaram produção significativa de IgG2 apenas contra MSP1a. As razões para as possíveis falhas de proteção são discutidas. Neste trabalho, é relatada a imunização de bovinos com MSP1a e MSP2 recombinantes de A. marginale, associadas com alúmen e CpG ODN 2006, e posterior desafio com isolado heterólogo, bem como a avaliação da resposta imune gerada.