Depression of Neuronal Protein Synthesis Initiation by Protein Tyrosine Kinase Inhibitors

Abstract— Growth factors stimulate cellular protein synthesis, but the intracellular signaling mechanisms that regulate initiation of mRNA translation in neurons have not been clarified. A rate-limiting step in the initiation of protein synthesis is the formation of the ternary complex among GTP, eukaryotic initiation factor 2 (elF-2), and the initiator tRNA. Here we report that genistein, a specific tyrosine kinase inhibitor, decreases tyrosine kinase activity and the content of phosphotyrosine proteins in cultured primary cortical neurons. Genistein inhibits protein synthesis by >80% in a dose-dependent manner (10–80 μg/ml) and concurrently decreases ternary complex formation by 60%. At the doses investigated, genistein depresses tyrosine kinase activity and concomitantly stimulates PKC activity. We propose that a protein tyrosine kinase participates in the initiation of protein synthesis in neurons, by affecting the activity of elF-2 directly or through a protein kinase cascade.     

Role of Intercellular and Intracellular Communication by Nitric Oxide in Coupling of Muscarinic Receptors to Activation of Guanylate Cyclase in Neuronal Cells

Abstract: Muscarinic receptor-mediated cyclic GMP formation and release of nitric oxide (NO) (or a precursor thereof) were compared in mouse neuroblastoma N1E-115 cells. [³H]Cyclic GMP was assayed in cells prelabeled with [³H]guanine. Release of NO upon the addition of muscarinic agonists to unlabeled neuroblastoma cells (NO donor cells) was quantitated indirectly by its ability to increase the [³H]cyclic GMP level in labeled cells whose muscarinic receptors were inactivated by irreversible alkylation (NO detector cells). Carbachol increased NO release in a concentration-dependent manner, with half-maximal stimulation at 173 μ M (compared to 96 μ M for direct activation of cyclic GMP formation). The maximal effect of carbachol in stimulating release of NO when measured indirectly was lower than that in elevating [³H]cyclic GMP directly in donor cells. Hemoglobin was more effective in blocking the actions of released NO than in attenuating direct stimulation of [³H]cyclic GMP synthesis. There was a good correlation between the ability of a series of muscarinic agonists to release NO or to activate [³H]cyclic GMP formation directly, and the potency of pirenzepine in inhibiting the two responses. Furthermore, there was a similar magnitude of desensitization of both responses by prolonged receptor activation or stimulation of protein kinase C. NO release was also regulated in relation to the cellular growth phase. A model is proposed in which a fraction of NO generated upon receptor activation does not diffuse extracellularly and stimulates cyclic GMP synthesis within the same cell where it is formed (locally acting NO). The remainder of NO that is extruded extracellularly might travel to neighboring cells (neurotransmitter NO) or might be taken back into the cells of origin (homing NO).     

川西峨眉晚白垩世夹关期河流沉积中的痕迹化石群落*

晚白垩世夹关组中的痕迹化石群落至少由12个痕迹属17个痕迹种组成,其中包括5个新痕迹种,即Cystichrtium cuwatitivum,Steinichnus laryus,Paradidymaulichnus emeiertsis,Monmorphichnus lineates和Rusophycus univalvis.这一化石群落主要是无脊椎动物的进食迹、觅食迹、爬迹、停息迹和居住迹,其中多数呈下浮痕和全浮痕保存,并形成于经常干旱的河流环境(大多出现在泛滥平原沉积中).该群落可识别出两个痕迹化石组合,即1) Scoyenia-Steinichnus-Rusophycus组合,它主要产自泛滥平原和漫滩环境;2)Skolithos-Arenicolites组合,它代表一种水道砂坝或曲流砂坝环境.     

Rates of apparent photosynthesis,respiration and dry matter accumulation in maize canopies

The rates of canopy apparent photosynthesis (P_C) and canopy respiration (R_c) were studied during vegetation season in two erectophile and two planophile hybrids of maize (Zea mays L.) grown at two canopy densities [7.5 plants m^-2 (HD) and 4.5 plants m^-2 (LD)]. Large differences in P_C, R_c, R_C/P_C, leaf area index (LAI), dry matter accumulation and grain yield were found among hybrids and plant densities. Variations in P_C and R_C were associated mainly with changes in LAI. There was also found change in P_C per unit LAI with time. The average R_C/P_C was 28.9 % for all treatments throughout the vegetation season. P_C and R_C per unit dry matter were higher in LD than in HD and decreased throughout the measurement period. The HD stand had higher P_C and yield in hybrids with erectophile foliage, whereas LD stand had higher P_C after male tetrad and got higher yield in hybrids with planophile foliage. Only R_C in hybrids of the two foliage types was higher under HD than under LD throughout the vegetation period.     

Cultivation of mammalian cells as aggregates in bioreactors: effect of calcium concentration of spatial distribution of viability

Recombinant human kidney epithelial 293 cells were cultivated as aggregates in suspension. The concentration calcium ion, in the range of 100 muM to 1mM, affected the rate of aggregate formation. During the course of cultivation the size distribution of aggregates shifted and the fraction of larger aggregates increased. This effect was more profound in cultures with a high calcium concentration. Scanning and transmission microscopic examination of the aggregates revealed that cell packing was greater in the high calcium cultures and that ultrastructural integrity was retained in aggregates from both low and high calcium cultures. Confocal microscopy was applied to examine the viability of cells in the interior of the aggregates. High viability was observed in the aggregates obtained from exponentially growing cultures. Aggregates from the high calcium culture in the stationary phase exhibited lower viability in the interior. With its ease of retention in a perfusion bioreactor, aggregate cultures offer an alternative choice for large-scale operation. (c) 1993 John Wiley & Sons, Inc.     

恒河猴、懒猴和树鼩的短时空间记忆功能与前额叶背侧部进化水平的相关性(英文)

本文研究探讨了进化地位不同的三种动物的短时空间记忆功能及其与前额叶背侧部进化水平的相关性。结果表明,在延缓反应作业中,经1000次训练后,7只恒河猴对空间位置的记忆时间平均为7.7±3.2min,懒猴为3.8±0.44min,而树鼩即使在延缓时间几乎为零秒的延缓反应中,其正确反应率也未达到90％标准。一种延缓时间仅测试一个单元,即不经训练的实验表明,恒河猴在延缓期为“0”—5min的各测试单元中,正确反应率稳定在80％以上;懒猴在延缓时间为“0”—4min的各测试单元中,平均正确反应率与恒河猴无明显差异,而当延缓时间增加到5min时,在延缓反应作业中取得的成绩显著下降;树鼩在延缓时间为1—5min的作业中取得的正确反应率在70％以下。3种动物在视觉辨别学习作业中却无明显差异。形态学研究表明,灵长类大脑前额叶的面积和结构的复杂性在进化过程中逐渐增大,如恒河猴大脑前额叶的表面积占大脑半球表面积的11.5％(Brodmann,1929),其内颗粒层发达,背侧部明显凸起,主沟区发达;懒猴的前额叶表面积占其大脑半球表面积的8.3％,背侧部凸起不显著,主沟未形成,额极内颗粒层分化明显,背侧部的内颗粒层较内侧部的发达程度差(Sanides,1967);树鼩的前额叶表面积占7.5％,额极的内颗粒层分化不明显,为非颗粒化区,此区之后为颗粒区     

Cloning of a novel, ubiquitously expressed human phosphatidylinositol 3-kinase and identification of its binding site on p85.

Phosphatidylinositol 3-kinase (PI 3-kinase) has been implicated as a participant in signaling pathways regulating cell growth by virtue of its activation in response to various mitogenic stimuli. Here we describe the cloning of a novel and ubiquitously expressed human PI 3-kinase. The 4.8-kb cDNA encodes a putative translation product of 1,070 amino acids which is 42% identical to bovine PI 3-kinase and 28% identical to Vps34, a Saccharomyces cerevisiae PI 3-kinase involved in vacuolar protein sorting. Human PI 3-kinase is also similar to Tor2, a yeast protein required for cell cycle progression. Northern (RNA) analysis demonstrated expression of human PI 3-kinase in all tissues and cell lines tested. Protein synthesized from an epitope-tagged cDNA had intrinsic PI 3-kinase activity and associated with the adaptor 85-kDa subunit of PI 3-kinase (p85) in intact cells, as did endogenous human PI 3-kinase. Coprecipitation assays showed that a 187-amino-acid domain between the two src homology 2 domains of p85 mediates interaction with PI 3-kinase in vitro and in intact cells. These results demonstrate the existence of different PI 3-kinase isoforms and define a family of genes encoding distinct PI 3-kinase catalytic subunits that can associate with p85.     

脂肪酶水解植物油的研究

本文报导了薄层层析法及高压液相色谱法定性定量地测定植物油甘油酯的组成,对不同来源的脂肪酶对植物油的水解进行了研究。实验结果表明,不同来源的脂肪酶对植物油(橄榄油)的水解性不同,同一脂肪酶水解不同种类的植物油,脂肪酶的水解率也不同,脂肪酶水解植物油有最适pH和最适温度。     

平菇子实体钙调素的分离纯化及其与猪脑钙调素的生物活性比较

平菇萃取液经酸性沉淀、热变性、Phenyl-Sepharose CL-4B疏水亲和层析和DEAE-Cellulose 52离子交換层析分冉出了电泳纯的真菌钙调素。在比较钙调素对磷酸二酯酶活化的能力和ELISA实验的免疫亲和力对发现,平菇钙调素与猪脑钙调素的生物活性有较大差异,提示在钙调素定量测定中有必要考虑到标准钙调素与样品钙调素之间的同源性差异。     

中国优质水果资源的分布与适宜生态环境

根据农业部在80年代两次组织评选出的全国189个优质水果产地的生态环境资料,用微型电子计算机系统建立数据库,统计分析柑桔、苹果和梨优质产品的构成比例、产区分布地域及其适宜的环境指标和主栽品种的生态适应性,为果树良种区域化栽培与选育提供依据。