Gene Expression of Neuropilin-1 and Its Receptors,VEGF/Semaphorin 3a,in Normal and Cancer Cells

Extracellular domains of the transmembrane glycoprotein, neuropilin-1 (Np1), specifically bind an array of factors and co-receptors including class-3 semaphorins (Sema3a), vascular endothelial growth factor (VEGF), hepatocyte growth factor, platelet-derived growth factor BB, transforming growth factor-β 1 (TGF-β1), and fibroblast growth factor2 (FGF2). Np1 may have a role in immune response, tumor cell growth, and angiogenesis, but its relative expression in comparison to its co-primary receptors, VEGF and Sema3a, is not known. In this study we determined the mRNA expression of Np1 and its co-receptors, VEGF and Sema3a, and the ratio of VEGF/Sema3a in different human and rodent cell lines. Expression of Np1, VEGF and Sema3a is very low in cells derived from normal tissues, but these proteins are highly expressed in tumor-derived cells. Furthermore, the ratio of VEGF/Sema3a is highly variable in different tumor cells. The elevated mRNA expression of Np1 and its putative receptors in tumor cells suggests a role for these proteins in tumor cell migration and angiogenesis. As different tumor cells exhibit varying VEGF/Sema3a ratios, it appears that cancer cells show differential response to angiogenic factors. These results bring to light the individual variation among the cancer-related genes, Np1, VEGF, and Sema3a, and provide an important impetus for the possible personalized therapeutic approaches for cancer patients.     

Structure of the ribosome associating GTPase HflX

The HflX‐family is a widely distributed but poorly characterized family of translation factor‐related guanosine triphosphatases (GTPases) that interact with the large ribosomal subunit. This study describes the crystal structure of HflX from Sulfolobus solfataricus solved to 2.0‐Å resolution in apo‐ and GDP‐bound forms. The enzyme displays a two‐domain architecture with a novel “HflX domain” at the N‐terminus, and a classical G‐domain at the C‐terminus. The HflX domain is composed of a four‐stranded parallel β‐sheet flanked by two α‐helices on either side, and an anti‐parallel coiled coil of two long α‐helices that lead to the G‐domain. The cleft between the two domains accommodates the nucleotide binding site as well as the switch II region, which mediates interactions between the two domains. Conformational changes of the switch regions are therefore anticipated to reposition the HflX‐domain upon GTP‐binding. Slow GTPase activity has been confirmed, with an HflX domain deletion mutant exhibiting a 24‐fold enhanced turnover rate, suggesting a regulatory role for the HflX domain. The conserved positively charged surface patches of the HflX‐domain may mediate interaction with the large ribosomal subunit. The present study provides a structural basis to uncover the functional role of this GTPases family whose function is largely unknown. Proteins 2010. © 2009 Wiley‐Liss, Inc.     

A cell-based screen for inhibitors of protein folding and degradation

Cancer cells are exposed to external and internal stresses by virtue of their unrestrained growth, hostile microenvironment, and increased mutation rate. These stresses impose a burden on protein folding and degradation pathways and suggest a route for therapeutic intervention in cancer. Proteasome and Hsp90 inhibitors are in clinical trials and a 20S proteasome inhibitor, Velcade, is an approved drug. Other points of intervention in the folding and degradation pathway may therefore be of interest. We describe a simple screen for inhibitors of protein synthesis, folding, and proteasomal degradation pathways in this paper. The molecular chaperone-dependent client v-Src was fused to firefly luciferase and expressed in HCT-116 colorectal tumor cells. Both luciferase and protein tyrosine kinase activity were preserved in cells expressing this fusion construct. Exposing these cells to the Hsp90 inhibitor geldanamycin caused a rapid reduction of luciferase and kinase activities and depletion of detergent-soluble v-Src::luciferase fusion protein. Hsp70 knockdown reduced v-Src::luciferase activity and, when combined with geldanamycin, caused a buildup of v-Src::luciferase and ubiquitinated proteins in a detergent-insoluble fraction. Proteasome inhibitors also decreased luciferase activity and caused a buildup of phosphotyrosine-containing proteins in a detergent-insoluble fraction. Protein synthesis inhibitors also reduced luciferase activity, but had less of an effect on phosphotyrosine levels. In contrast, certain histone deacetylase inhibitors increased luciferase and phosphotyrosine activity. A mass screen led to the identification of Hsp90 inhibitors, ubiquitin pathway inhibitors, inhibitors of Hsp70/Hsp40-mediated refolding, and protein synthesis inhibitors. The largest group of compounds identified in the screen increased luciferase activity, and some of these increase v-Src levels and activity. When used in conjunction with appropriate secondary assays, this screen is a powerful cell-based tool for studying compounds that affect protein synthesis, folding, and degradation.     

The alternative pathway in cucumber seedlings under low temperature stress was enhanced by salicylic acid

The alternative pathway is a cyanide-resistant and non-phosphorylatory electron transport pathway in mitochondria of higher plants. Alternative oxidase (AOX) is the terminal oxidase of this pathway. Our present study investigated the effect of exogenous salicylic acid (SA) on alternative pathway in cucumber (Cucumis sativus L.) seedlings under low temperature stress. Results showed that during the process of low temperature stress, the alternative pathway capacity was enhanced as AOX expression increased in SA pretreated seedlings. Compared with seedlings without SA pretreatment, slower decrease of relative water content and lower levels of electrolyte leakage, H₂O₂ and malonyldialdehyde content were detected in SA pretreated seedlings. These results indicated that SA could alleviate the injury caused by low temperature on cucumber seedlings. Since the special protective functions of alternative pathway and AOX in plants, we suggested that the alternative pathway was related to SA-mediated plant resistance to environmental stresses such as low temperature.     

泰顺杜鹃扦插繁殖试验研究

本研究采用插床育苗、薄膜覆盖的方法对泰顺杜鹃插穗进行不同剂型生根粉、不同浸泡时间、不同年龄、不同插穗长度和不同品种对比的扦插实验。结果表明:(1)生长调节剂GGR 6号相对于萘乙酸、清水生根率和不定根数较多;(2)按照高浓度速蘸低浓度浸泡原则,同激素的GGR 6号在同一浓度100 mg/L进行浸泡,以1.5 h~2h效果显著;(3)对于不同年龄生的插穗,一年生、两年生和三年生的虽不存在显著差异,但一年生插穗生根性状指标还是略高于其他;(4)对于不同规格的插穗,长度6~10 cm的生根率明显好于其他规格;(5)不同品种杜鹃生根状况存在差异,实验结果刺毛杜鹃整体性状优于泰顺杜鹃。     

人工接种猪瘟病毒对猪外周血白细胞的影响

为研究CSFV强毒感染对猪外周血白细胞的影响,本研究用猪瘟病毒石门株(CSFV SM)感染60日龄仔猪后,分析外周血中CSFV核酸载量动态变化、白细胞亚群变化和白细胞SLAⅠ和SLAⅡDR分子的表达情况。实验结果显示:实验仔猪经CSFV感染后48小时体温升高并可以在血液中检测到CSFV核酸,核酸载量持续升高,在感染后6日(DPI)达到最大值,为2 DPI时核酸载量的104.84±0.98倍;WBC、LYM、PLT数量持续降低,WBC在1DPI和2DPI分别降至65.87%和50.00%,LYM在1~3DPI分别降至70.68%、47.88%和23.29%,PLT数量持续降低,6DPI时仅为初始值的34.59%;NK、γδT、Tc、Th、CD3+CD4+CD8+和CD3-CD4-CD8-淋巴细胞在感染后均不同程度的减少,其中NK细胞在1DPI时减少78.49%,而后变化与1DPI比较差异不显著,γδT、Tc、CD3-CD4-CD8-、CD3+CD4+CD8+在3DPI时分别降至41.74%、43.83%、15.87%和32.96%,Th细胞在感染后持续下降,在6DPI时减少至42.95%;感染后淋巴细胞中表达S...     

工业园区磷代谢分析——以江苏宜兴经济开发区为例

工业园区是工业活动的重要载体,也是水污染控制与治理的焦点对象。运用物质代谢分析方法解析水中主要污染主导元素在工业园区的代谢途径、结构与动力机制,有助于寻求提高水资源利用效率和减缓水环境污染压力的举措。以江苏宜兴经济开发区为案例,基于物质流分析方法构建了工业园区的磷代谢网络,详细解析了工业系统和污水处理模块的磷代谢途径和通量。研究表明,印染、食品加工和机械(磷化)行业是宜兴经济开发区的主要磷排放源;企业自备处理设施除磷效果不佳,磷去除率大约为60%,集中污水处理厂可以有效除磷,去除率约75%;生活污水磷去除率低;不经处理直接排入水体的降水给水体造成了较大的负荷,为34%。由此,建议企业完善简单的处理设施,污水处理厂提高企业纳管率,同时园区加强对生活污水、生活垃圾和企业固体排放物的管理。     

水鼩鼱消化道5-羟色胺阳性细胞的形态与分布

应用免疫酶标技术ABC( avidin-biotin-peroxidase complex)法,对水鼩鼱(Neomys fodiens)消化道内5-羟色胺(5-HT)阳性细胞的分布及形态进行研究.结果显示,5-羟色胺阳性细胞仅在肠内分布,食管和胃处没有分布.细胞分布密度曲线呈抛物线型,空肠部分布密度最高,十二指肠部次之...