Molecular characterization and subcellular localization of Carassius auratus interferon regulatory factor-1

Interferon (IFN)-regulatory transcription factor-1 (IRF-1) has been studied in mammals and fish but little is known about the relationship between its gene structure and nuclear 'ion of IRF-1 protein. In this study, a cDNA encoding Carassius auratus IRF-1 (CaIRF-1) was isolated from an interferon-producing cell line, C. ouratus blastulae embryonic (CAB) cells, exposed to UV-inactivated grass carp hemorrhagic virus (GCHV). The CaIRF-1 genomic locus exhibits exon-intron arrangements similar to those of other vertebrate IRF-1 loci, with nine exons and eight introns, although together with pufferfish IRF-1, CaIRF-1 distinguishes itself from other vertebrate IRF-1 genes by a relatively compact genomic size. Similar to the known IRF-1 genes, CaIRF-1 is ubiquitously expressed, and is upregulated in vitro and in vivo in response to virus, Poty I:C, or CAB INF-containing supernatant (ICS). Subcellular localization analysis confirms the nuclear distribution of CaIRF-1 protein, and reveals two nuclear localization signals (NILS), any one of which is sufficient for nuclear translocation of CaIRF-1. One NLS Locates to amino acids 117-146, and appears to be the structural and functional equivalent of the NLS in mammalian IRF-1. The second NLS (amino acids 73-115) is found within the DNA-binding domain (DBD) of CaIRF-1, and contains two regions rich in basic amino acids (''(KDKSINK101)-K-95" and ''(75)KTWKANFR(82)"). In comparison with mammalian IRF-1, in which the corresponding amino acid stretch does not seem to drive nuclear translocation, five conserved basic amino acids (K-75, K-78, R-82, K-95, and K-101) and one non-conserved basic amino acid (K-97) are present in this NLS from CaIRF-1. This observation suggests that K97 Of CaIRF-1 might be essential for the function of its second NLS, wherein the six basic aminoacids might cooperate to drive CaIRF-1 to the nucleus. Therefore, the current study has revealed a new nuclear localization motif in the DBD of a vertebrate IRF-1. (C) 2007 Elsevier Ltd. All rights reserved.

Identification and characterization of two homologues of interferon-stimulated gene ISG15 in crucian carp

ISG15 is one of the most strongly induced genes upon viral infection, interferon (IFN) stimulation, and lipopolysaccharide, (LPS) stimulation, and only one copy has been found in mammals so far. Here two fish ISG15 genes, termed CaISG15-1 and CaISG15-2, have been cloned and sequenced from UV-inactivated GCHV (grass carp haemorrhagic virus)-infected and IFN-produced CAB cells (crucian carp Carassius auratus blastulae embryonic cells) by suppression subtractive hybridization. The full-length cDNA sequences of two crucian carp ISG15 encode a 155-amino-acid protein and a 161-amino-acid protein, both of which show 78.9% identity overall and possess the characteristic structures of mammalian ISG15 proteins including two tandem ubiquitin-like domains and the C-terminal canonical LRLRGG motif. In CAB cells treated with different stimuli including active virus, UV-inactivated GCHV and IFN containing supernatant (ICS), the expression of both CaISG15-1 and CaISG15-2 was up-regulated but displayed different kinetics. Poly I:C and LPS were also able to induce an increase in mRNA for both genes. In CAB cells responsive to active GCHV, UV-inactivated GCHV, CAB ICS, Poly 1:12 and LPS, CaISG15-1 was upregulated more significantly than CaISG15-2. These results suggest that there are two ISG15 homologues in crucian carp, both of which might play distinct roles in innate immunity against viral and bacterial infection. (c) 2006 Elsevier Ltd. All rights reserved.

Identification of two novel interferon-stimulated genes from cultured CAB cells induced by UV-inactivated grass carp hemorrhage virus

Interferon (IFN) exerts its antiviral effect by inducing the expression of a number of IFN-stimulated genes (ISGs) to establish a host antiviral state. Earlier studies identified some important fish IFN system genes from IFN-induced CAB cells (crucian carp Carassius auratus L. embryonic blastulae cells) after treatment with UV-inactivated GCHV (grass carp hemorrhage virus). Herein, the cloning of 2 novel IFN-stimulated genes, termed Gig1 and Gig2, is described for the same cell system. The complete cDNA sequences of Gig1 and Gig2 contain 1244 bp encoding for a 194-amino-acid protein and 693 bp for a 158-amino-acid protein, respectively. A search of public databases revealed that these are 2 novel IFN-stimulated genes, since neither significant homologous genes nor conserved motifs were identified. Active GCHV, UV-inactivated GCHV and CAB IFN-containing supernatant (ICS) induced transcription of these genes and distinct kinetics were observed. An analysis of differences in expression between the 2 genes and the IFN signal factors CaSTAT1 and CaIRF7 indicated that GCHV infection activated different signal pathways for their up-regulation. Upon virus infection, the transcription of Gig1 but not of Gig2 is strongly suppressed by cycloheximide (CHX). In contrast, following treatment with CAB IFN-containing supernatant, CHX does not inhibit either gene transcription. The results suggest that GCHV infection can induce expression of both Gig1 and Gig2 via newly synthesized CAB IFN, most probably through the JAK-STAT signal pathway, and can also directly activate Gig2 transcription without ongoing protein synthesis.

Nano-layer structure of silicon-on-insulator materials

Silicon-on-insulator (SOI) has been recognized as a promising semiconductor starting material for ICs where high speed and low power consumption are desirable, in addition to its unique applications in radiation-hardened circuits. In the present paper, three novel SOI nano-layer structures have been demonstrated. ULTRA-THIN SOI has been fabricated by separation by implantation of oxygen (SIMOX) technique at low oxygen ion energy of 45 keV and implantation dosage of 1.81017/cm2. The formed SOI layer is uniform with thickness of only 60 nm. This layer is of crystalline quality. and the interface between this layer and the buried oxide layer is very sharp, PATTERNED SOI nanostructure is illustrated by source and drain on insulator (DSOI) MOSFETs. The DSOI structure has been formed by selective oxygen ion implantation in SIMOX process. With the patterned SOI technology, the floating-body effect and self-heating effect, which occur in the conventional SOI devices, are significantly suppressed. In order to improve the total-dose irradiation hardness of SOI devices, SILICON ON INSULATING MULTILAYERS (SOIM) nano-structure is proposed. The buried insulating multilayers, which are composed of SiOx and SiNy layers, have been realized by implantation of nitride and oxygen ions into silicon in turn at different ion energies, followed by two steps of high temperature annealing process, respectively, Electric property investigation shows that the hardness to the total-dose irradiation of SOIM is remarkably superior to those of the conventional SIMOX SOI and the Bond-and-Etch-Back SOI.

Space grown semi-insulating gallium arsenide single crystal and its application

Low noise field effect transistors and analogue switch integrated circuits (ICs) have been fabricated in semi-insulating gallium arsenide (SI-GaAs) wafers grown in space by direct ion-implantation. The electrical behaviors of the devices and the ICs have surpassed those fabricated in the terrestrially grown SI-GaAs wafers. The highest gain and the lowest noise of the transistors made from space-grown SI-GaAs wafers are 22.8 dB and 0.78 dB, respectively. The threshold back-gating voltage of the ICs made from space-grown SI-GaAs wafers is better than 8.5 V The con-elation between the characterizations of materials and devices is studied systematically. (C) 2002 COSPAR. Published by Elsevier Science Ltd. All rights reserved.

固斑蝰蛇毒C-型凝集素样蛋白Dabocetin及L-氨基酸氧化酶DRS-LAO的研究

本论文研究了从圆斑蛙蛇泰国亚种（Daboia russellii siamensis）蛇毒中纯化的C一型凝集素样蛋白Dabocetin和L一氨基酸氧化酶DRS一AO的理化性质、生物学活性和分子克隆。Dabocetin是分子量约为28扔。的异二聚体蛋白，它由分子量约为15.0kDa和14．5kDa的两个同源亚基以和p共价结合形成。N－末端氨基酸序列比较显示，Dabocetin与目前已知的蛇毒c一型凝集素样蛋白有很高的同源性。即使在终浓度达50．0。叫而时，Dabocetin也不能直接诱导血小板聚集。此外，在终浓度为40.00μg/ml时，Dabocetin几乎不能抑制由AdP，TMVA和stejnulxin诱导的血小板聚集。但是，Dabocetin呈剂量依赖地抑制瑞斯托霉素诱导的血小板凝集，其半数抑制率ICS。值为10.80ug/ml。流式细胞仪分析表明，Dabocetin显著抑制单克隆抗体522与GPIba的结合，提示Dabocetin很可能是一个GPIb结合蛋白。从圆斑蛙蛇的毒腺中克隆到了7个编码不同蛇毒C一型凝集素样蛋白亚基的七DNA（命名为DRs一1至DRs一7）。其中，DRsLS编码Dabocetin的a亚基，DRS一6编码Dabocetin的p亚基。DRs一1和DRS一2很可能是圆斑蛙蛇毒腺中表达的X因子激活剂的两条轻链LCZ和LCI的山NA。DRS一3，DRS毛4和DRSL7可能是圆斑蛙蛇毒腺中表达的C一型凝集素样蛋白p亚基的。NA。DRsLAO是一个新的L一氨基酸氧化酶，比活力为1．98U加噶。十二烷基磺酸钠一聚丙烯酞氨凝胶电泳（SDs-PAGE）分析显示，该酶在还原和非还原条件下均呈现一条蛋白带，表观分子量约为58kDa。N-末端氨基酸序列比较显示，DRS一AO与目前已知的蛇毒L一氨基酸氧化酶有很高的同源性。该酶的最适底物为L一亮氨酸，最适pH为8．8。DRs一Ao呈剂量依赖地抑制扔P和仆IvA诱导的血小板聚集，其半数抑制率ICS。值分别为32.8μg/ml和32.3μg/ml。DRS-LAO对金黄色葡萄球菌（灯Cc25923）和耐甲氧西林金黄色葡萄球菌有较强的抗菌作用。DRs一AO对金黄色葡萄球菌必Tcc25923）的最低抑菌浓度卿C）和最低杀菌浓度耐甲氧西林金黄色葡萄球菌的孤CS。和呱Cg。值分别为18．。林留时和36.0μg/ml；DRSLAo对耐甲氧西林金黄色葡萄球菌的MBC50和MBCg。值分别为36.0μg/ml和72.0μg/ml。通过对DRS一AO的分子克隆，得到了编码DRS-AO的部分cDNA序列。

Nano-layer structure of silicon-on-insulator materials

Silicon-on-insulator (SOI) has been recognized as a promising semiconductor starting material for ICs where high speed and low power consumption are desirable, in addition to its unique applications in radiation-hardened circuits. In the present paper, three novel SOI nano-layer structures have been demonstrated. ULTRA-THIN SOI has been fabricated by separation by implantation of oxygen (SIMOX) technique at low oxygen ion energy of 45 keV and implantation dosage of 1.81017/cm2. The formed SOI layer is uniform with thickness of only 60 nm. This layer is of crystalline quality. and the interface between this layer and the buried oxide layer is very sharp, PATTERNED SOI nanostructure is illustrated by source and drain on insulator (DSOI) MOSFETs. The DSOI structure has been formed by selective oxygen ion implantation in SIMOX process. With the patterned SOI technology, the floating-body effect and self-heating effect, which occur in the conventional SOI devices, are significantly suppressed. In order to improve the total-dose irradiation hardness of SOI devices, SILICON ON INSULATING MULTILAYERS (SOIM) nano-structure is proposed. The buried insulating multilayers, which are composed of SiOx and SiNy layers, have been realized by implantation of nitride and oxygen ions into silicon in turn at different ion energies, followed by two steps of high temperature annealing process, respectively, Electric property investigation shows that the hardness to the total-dose irradiation of SOIM is remarkably superior to those of the conventional SIMOX SOI and the Bond-and-Etch-Back SOI.

Space grown semi-insulating gallium arsenide single crystal and its application

Low noise field effect transistors and analogue switch integrated circuits (ICs) have been fabricated in semi-insulating gallium arsenide (SI-GaAs) wafers grown in space by direct ion-implantation. The electrical behaviors of the devices and the ICs have surpassed those fabricated in the terrestrially grown SI-GaAs wafers. The highest gain and the lowest noise of the transistors made from space-grown SI-GaAs wafers are 22.8 dB and 0.78 dB, respectively. The threshold back-gating voltage of the ICs made from space-grown SI-GaAs wafers is better than 8.5 V The con-elation between the characterizations of materials and devices is studied systematically. (C) 2002 COSPAR. Published by Elsevier Science Ltd. All rights reserved.

全地形轮式移动机器人运动学建模与分析

提出全地形轮式移动机器人的正逆运动学问题。将机器人看成一个混合串-并联多刚体系统,从每个轮-地接触点到机器人车体分别构成一个串联子系统,抛弃车轮纯滚动假设,在轮-地接触点处建立瞬时坐标系,考虑车轮的平面滑移,从而对每个串联子系统形成一个封闭的速度链。对于每个速度闭链,可直接在驱动轮轮心处写出从机器人各驱动轮到机器人本体之间的运动方程,将每个速度闭链的运动方程合并即可得到机器人的整体运动学模型。以一个具有被动柔顺机构的六轮全地形移动机器人为对象进行推导,该方法既考虑了地形不平的影响,又考虑了车轮的前向、侧向及转向滑移,已知机构参数后就可以直接写出机器人的速度方程,且便于运动学求解。该方法对于轮式移动机器人的运动学建模具有一般性,且具有物理意义明确、推导过程简洁等特点。

基于改进SICA的人脸特征提取方法研究

独立分量分析是一种有效的人脸特征提取方法。考虑到人脸样本的对称性,本文采用对称独立分量分析的方法对人脸样本进行特征提取。为了提高独立分量分析法表征人脸特征空间的能力,本文采用遗传算法对特征空间进行选择优化,获得最优的人脸特征子集。仿真实验表明,本文提出方法的识别率明显的好于独立分量分析方法的识别率。

天山乌鲁木齐河源1号冰川成冰过程的地球化学研究--以低分子有机酸为主

冰川上的表面雪层是联系大气成分与冰芯记录的重要纽带，对雪－冰现代过程的研究将有助于正确解释保存在冰芯中的古气候和环境信息。山地冰川雪冰中有机酸记录的研究在认识有机酸生物地球化学循环方面占很大的优势，本文依托天山冰川观测试验站开展的冰雪物理、化学现代过程方面的研究，对乌鲁木齐河源1号冰川中低分子有机酸和无机阴离子在成冰作用过程中的迁移转化进行了研究。新雪和保存较好的表层雪主要用来分析采样点低分子有机酸和无机阴离子的季节变化特征，粒雪坑样品主要用于进行低分子有机酸和无机阴离子沉积后过程的研究。 雪冰样品采集于1号冰川东支东经86°49′，北纬43°06′，海拔4 130 m 处，2004年3月至2005年3月期间，共获取了23个连续雪坑剖面的粒雪样品。采用美国Dionex公司的ICS-90型离子色谱仪加RFC-30型淋洗液在线发生器进行测定，检测到的低分子有机酸主要有HCOO-、CH3COO-、C2H5COO- 和(COO)22-，无机阴离子主要有F-、Cl-、NO2-、NO3-、SO42- 和PO43-。 表层雪是研究粒雪化和成冰作用过程中化学组成变化的起点。为配合天山乌鲁木齐河源1号冰川沉积后过程中化学组成变化的研究，我们首先对表层雪样品（雪坑表层5cm的雪样）进行了低分子有机酸和无机阴离子含量的分析。结果显示，除(COO)22- 外，大部分有机酸和高浓度的无机阴离子因受到周围环境和盛行风的影响呈现出明显的季节变化特征，即夏半年离子浓度变化剧烈，最大值和最小值同时出现在夏半年，冬半年的浓度则相对小而稳定；而(COO)22- 和低浓度的无机阴离子随季节变化的特征不明显，在全年均显示出波动性。数据表明，在外界条件不变的情况下，表层雪可以长时间（至少半年时间）保存其中高含量的化学组成不被改变。 通过对高含量低分子有机酸和高含量无机阴离子的沉积后过程的研究，发现它们在沉积后的迁移转化过程受温度和融水的影响较大，在不同时期表现出不同的季节变化特征。雪层消融初期，淋溶作用导致雪层内的大部分离子组分进入初始融水中，并随着融水下渗、聚集，使下部雪层中离子组分的浓度不断增加；当消融进一步加剧，大部分低分子有机酸和无机阴离子随融水径流而流失，雪层中记录的低分子有机酸和无机阴离子含量迅速降低；在消融季节，峰值向雪坑底部移动的速度较快。在冬半年的负温条件下，雪层中记录的低分子有机酸和无机阴离子的含量及其保存在雪坑下部的峰值都相对稳定，这也再次说明如果外界条件不发生改变，雪冰可以长时间（至少半年时间）保存其中的化学组成不被改变这个结论；另外，冬半年雪坑上部记录的峰值在一定程度上可以反映它们在大气中的初始水平。CH3COO- 和(COO)22- 的峰值通常会出现在污化层的附近，与污化层的位置具有较好的一致性，说明污化层对CH3COO- 和(COO)22- 的沉积后过程可能有一定的影响。

Immunization with cocktail of HIV-derived peptides in montanide ISA-51 is immunogenic, but causes sterile abscesses and unacceptable reactogenicity.

BACKGROUND: A peptide vaccine was produced containing B and T cell epitopes from the V3 and C4 Envelope domains of 4 subtype B HIV-1 isolates (MN, RF, CanO, & Ev91). The peptide mixture was formulated as an emulsion in incomplete Freund's adjuvant (IFA). METHODS: Low-risk, healthy adult subjects were enrolled in a randomized, placebo-controlled dose-escalation study, and selected using criteria specifying that 50% in each study group would be HLA-B7+. Immunizations were scheduled at 0, 1, and 6 months using a total peptide dose of 1 or 4 mg. Adaptive immune responses in16 vaccine recipients and two placebo recipients after the 2nd immunization were evaluated using neutralization assays of sera, as well as ELISpot and ICS assays of cryopreserved PBMCs to assess CD4 and CD8 T-cell responses. In addition, (51)Cr release assays were performed on fresh PBMCs following 14-day stimulation with individual vaccine peptide antigens. RESULTS: 24 subjects were enrolled; 18 completed 2 injections. The study was prematurely terminated because 4 vaccinees developed prolonged pain and sterile abscess formation at the injection site-2 after dose 1, and 2 after dose 2. Two other subjects experienced severe systemic reactions consisting of headache, chills, nausea, and myalgia. Both reactions occurred after the second 4 mg dose. The immunogenicity assessments showed that 6/8 vaccinees at each dose level had detectable MN-specific neutralizing (NT) activity, and 2/7 HLA-B7+ vaccinees had classical CD8 CTL activity detected. However, using both ELISpot and ICS, 8/16 vaccinees (5/7 HLA-B7+) and 0/2 controls had detectable vaccine-specific CD8 T-cell responses. Subjects with moderate or severe systemic or local reactions tended to have more frequent T cell responses and higher antibody responses than those with mild or no reactions. CONCLUSIONS: The severity of local responses related to the formulation of these four peptides in IFA is clinically unacceptable for continued development. Both HIV-specific antibody and T cell responses were induced and the magnitude of response correlated with the severity of local and systemic reactions. If potent adjuvants are necessary for subunit vaccines to induce broad and durable immune responses, careful, incremental clinical evaluation is warranted to minimize the risk of adverse events. TRIAL REGISTRATION: ClinicalTrials.gov NCT00000886.

Safety and immunogenicity of a replication-defective adenovirus type 5 HIV vaccine in Ad5-seronegative persons: a randomized clinical trial (HVTN 054).

BACKGROUND: Individuals without prior immunity to a vaccine vector may be more sensitive to reactions following injection, but may also show optimal immune responses to vaccine antigens. To assess safety and maximal tolerated dose of an adenoviral vaccine vector in volunteers without prior immunity, we evaluated a recombinant replication-defective adenovirus type 5 (rAd5) vaccine expressing HIV-1 Gag, Pol, and multiclade Env proteins, VRC-HIVADV014-00-VP, in a randomized, double-blind, dose-escalation, multicenter trial (HVTN study 054) in HIV-1-seronegative participants without detectable neutralizing antibodies (nAb) to the vector. As secondary outcomes, we also assessed T-cell and antibody responses. METHODOLOGY/PRINCIPAL FINDINGS: Volunteers received one dose of vaccine at either 10(10) or 10(11) adenovector particle units, or placebo. T-cell responses were measured against pools of global potential T-cell epitope peptides. HIV-1 binding and neutralizing antibodies were assessed. Systemic reactogenicity was greater at the higher dose, but the vaccine was well tolerated at both doses. Although no HIV infections occurred, commercial diagnostic assays were positive in 87% of vaccinees one year after vaccination. More than 85% of vaccinees developed HIV-1-specific T-cell responses detected by IFN-γ ELISpot and ICS assays at day 28. T-cell responses were: CD8-biased; evenly distributed across the three HIV-1 antigens; not substantially increased at the higher dose; and detected at similar frequencies one year following injection. The vaccine induced binding antibodies against at least one HIV-1 Env antigen in all recipients. CONCLUSIONS/SIGNIFICANCE: This vaccine appeared safe and was highly immunogenic following a single dose in human volunteers without prior nAb against the vector. TRIAL REGISTRATION: ClinicalTrials.gov NCT00119873.

Furnishing a Globalized World: Local Distinctiveness in the International Furniture Industry

Advances in technology, communication, and transportation over the past thirty years have led to tighter linkages and enhanced collaboration across traditional borders between nations, institutions, and cultures. This thesis uses the furniture industry as a lens to examine the impacts of globalization on individual countries and companies as they interact on an international scale. Using global value chain analysis and international trade data, I break down the furniture production process and explore how countries have specialized in particular stages of production to differentiate themselves from competitors and maximize the benefits of global involvement. Through interviews with company representatives and evaluation of branding strategies such as advertisements, webpages, and partnerships, I investigate across four country cases how furniture companies construct strong brands in an effort to stand out as unique to consumers with access to products made around the globe. Branding often serves to highlight distinctiveness and associate companies with national identities, thus revealing that in today’s globalized and interconnected society, local differences and diversity are more significant than ever.

An investigation of assessment policy and practices in terms of broad social and educational purposes

Based upon relevant literature, this study investigated the assessment policy and practices for the BSc (Hons) Computing Science programme at the University of Greenwich (UOG), contextualising these in terms of broad social and educational purposes. It discusses Assessment, and then proceeds to give a critical evaluation of the assessment policy and practices at the UOG. Although this is one case study, because any of the features of the programme are generic to other programmes and institutions, it is of wider value and has further implications. The study was concluded in the summer of 2002. It concludes that overall, the programme's assessment policy and practices are well considered in terms of broad social and educational purposes, although it identifies and outlines several possible improvements, as well as raising some major issues still to be addressed which go beyond assessment practices.