Involvement of the Golgi region in the intracellular trafficking of cholera toxin

The intracellular pathway following receptor-mediated endocytosis of cholera toxin was studied using brefeldin A (BFA), which inhibited protein secretion and induced dramatic morphological changes in the Golgi region. In both mouse Y1 adrenal cells and CHO cells, BFA at 1 μg/ml caused a 80–90% inhibition of the cholera toxin (CT)-elevation of intracellular cAMP. The inhibition of the cytotoxicity of CT by BFA was also observed in a rounding assay of Y1 adrenal cells. The inhibition of CT cytotoxicity by BFA was dose dependent, with the ID₅₀ value similar to the LD₅₀ of BFA in Y1 adrenal cells. Binding and internalization of [¹²⁵I]-cholera toxin in Y1 adrenal cells was not affected by BFA. Unlike the BFA-sensitive cell lines such as Y1 adrenal and CHO cells, BFA at 1 μg/ml did not inhibit the cytotoxicity of CT in PtK₁ cells, of which the Golgi structure was BFA-resistant. These results strongly suggest that a BFA-sensitive Golgi is required for the protection of CT cytotoxicity by BFA. In contrast, elevation of the intracellular cAMP by forskolin, which acts directly on the plasma membrane adenylate cyclase, was not affected by BFA. These observations indicate that the intoxication of target cells by CT requires an intact Golgi region for its intracellular trafficking and/or processing. In this respect, CT shares a common intracellular pathway with ricin, Pseudomonas toxin, and modeccin, even though their structures and modes of action are very different. © 1993 Wiley-Liss, Inc.     

Assignment of human platelet GP2B (GPIIb) gene to chromosome 17, region q21.1-q21.3

Summary The platelet GPIIb-IIIa complex functions as a receptor for fibrinogen, fibronectin, and von Willebrand factor on activated platelets. This glycoprotein is a member of a broadly distributed family of structurally and immunologically related membrane receptors involved in cell-cell contact and cell-matrices interactions. GPIIb-IIIa is a heterodimer complex composed of GPIIb (the subunit), which consists of two disulfide-linked heavy and light chains, and GPIIIa (the subunit), which is a single polypeptide chain. Congenital absence of platelet GPIIb-IIIa in Glanzmann's thrombasthenia results in a severe bleeding disorder characterized by defective platelet aggregation and failure of fibrinogen to bind to platelets. The gene coding for GPIIb was located on 17q21.1-17q21.3 as determined by in situ hybridization with a 2650-pb GP2B (GPIIb) cDNA probe prepared from human megakaryocytes.     

营养膜无土栽培技术

前言营养膜技术(Nutrient Film Technique简称NFT)是国外广泛采用的一种特殊的无土栽培方法。它是将植物栽植在一个狭长的不透水的营养槽中,槽中有缓慢流动的营养液。植物根系的一部分浸没在营养液中,一部分露在空气中,植物从这种小环境中吸收水、营养物和氧气。由于这种方法采用很薄一层营养液在根际周围流动,裸露的根     

从抗体阳性的甲型肝炎接触者粪便中分离出一株甲型肝炎病毒

几年来,国内外已相继用组织培养法分离多株甲型肝炎病毒(HAV),并已证实了甲型肝炎患者及亚临床型感染在传染本病中的重要性。但HAV在流行点的正常人群中存在短期携带的问题还尚未见报道。本文采用人胚肺二倍体细胞(2BS)从甲型肝炎接触者粪便中分离出一株HAV,并经免疫电镜、免疫荧光及参比血清鉴定等证实。现将结果报告如下。     

ESR检测大鼠肺巨噬细胞释放的活性氧自由基

用ESR捕捉技术检测大鼠AM释放的活性氧自由基的性质表明:1.PMA和BCG均能刺激AM产生较强的OH·;能刺激人末稍血白细胞释放活性氧自由基的ConA和顺铂在本实验条件下未能使AM产生活性氧自由基信号。2.经膜活性剂PMA刺激的AM所释放活性氧自由基的高峰在刺激后2min,而经颗粒性物质BCG刺激,AM释放自由基的高峰时间明显后移。3.测试体系中的AM数过多或过少都不适合捕捉ESR信号。在本实验条件下,捕捉到最高自由基信号的AM终浓度为5×10⁷AM/ml。4.测试体系中存在DETAPAC或EDTA,可使捕捉到的自由基信号明显增强。     

滇产薄荷的化学研究

研究了滇产３８个薄荷样品,测定了样品的得油率及化学成分。滇产薄荷的得油率在０．１８％￣０．５２％之间。从挥发油中鉴定出了１００多种化学成分,主要含醇、酮、酯、萜烯类化合物。栽培的家薄荷挥发油富含香芹酮、柠檬烯,其化学分类属于香芹酮系列。野生薄荷挥发油富含薄荷醇和薄荷酮,属于薄荷酮系列;部分野薄荷样品,富含香芹酮、环氧辣薄荷烯酮或芳樟醇,属于混合系列。     

Artificial mosaic protein containing antigenic epitopes of hepatitis E virus.

A synthetic gene encoding an artificial polypeptide composed of antigenic epitopes of the hepatitis E virus (HEV) proteins was constructed from short oligodeoxyribonucleotides by using PCR. The polypeptide comprises a mosaic of three antigenically active dominant regions from the protein encoded by open reading frame 2 (ORF2), one antigenically active region from the protein encoded by ORF3 of the Burmese HEV strain, and one antigenically active region from the protein encoded by ORF3 of the Mexican HEV strain. The mosaic protein was expressed in Escherichia coli as a chimera with glutathione S-transferase or beta-galactosidase. Guinea pig sera containing antibodies to the corresponding HEV synthetic peptides were used to demonstrate by Western immunoblot analysis and enzyme immunoassay the presence and accessibility of all HEV-specific antigenic epitopes introduced into the mosaic protein. Both the glutathione S-transferase and beta-galactosidase hybrid proteins were analyzed by using a panel of human anti-HEV-positive and -negative sera. The data obtained strongly indicate a diagnostic potential for the mosaic protein.