Role of prostaglandins and cAMP in the secretory effects of cholera toxin

The role of adenosine 3',5'-monophosphate (cAMP) and prostaglandins in the pathogenesis of experimental cholera was evaluated. Fluid accumulated in the rabbit intestinal loop model after 16 hours of incubation with cholera toxin, prostaglandin E1, or prostaglandin E2, but not with membrane-permeable derivatives of cAMP or forskolin. Dibutyryl cAMP triggered a small, transient intestinal fluid accumulation response by 4.5 hours; however, the fluid was completely absorbed by 9 hours. After exposure of intestinal loops to cholera toxin, prostaglandin E was released into the intestinal lumen in a concentration-dependent manner independent of cAMP. Thus, not only cAMP, but also prostaglandins may regulate water and electrolyte secretion in cholera.     

Immune response to cholera toxin epitope inserted in Salmonella flagellin

Bacterial flagella are potent immunogens and aromatic-dependent (aro) Salmonella as live vaccines evoke humoral and cellular immune responses. Such strains expressing epitopes of protective antigens as inserts in flagellin would provide a novel way to vaccinate against diseases caused by unrelated pathogens. A synthetic oligonucleotide specifying an epitope of cholera toxin subunit B was inserted in a Salmonella flagellin gene. The chimeric flagellin functioned normally and the epitope was expressed at the flagellar surface. Parenteral administration to mice of an aro A flagellin-negative strain of S. dublin expressing the chimeric flagellin gene evoked antibody to cholera toxin.     

玉米对大斑病的抗性与叶组织细胞膜耐毒力的关系研究

用真菌毒素处理玉米叶片的方法,研究玉米品种对大斑病的抗性。结果证实,大斑病菌毒素对玉米叶片有很强的毒性,造成叶组织细胞膜损伤,损伤程度与玉米品种的抗性程度有关。不具Ht基因型的感病品种的叶组织细胞膜损伤程度随着毒素处理时间的延长而加重,但抗病品种叶组织细胞膜的损伤程度不明显。Ht基因型的玉米品种叶组织细胞膜对真菌毒素的敏感性高于不具Ht基因型的品种,其叶组织在毒素作用下很快枯萎,限制了体内菌丝体的进一步扩展,从而对大斑病产生高抗。     

Electrical coupling: low resistance junctions between mitotic and interphase fibroblasts in tissue culture

Dividing cells of chick embryonic fibroblasts and of mouse embryonic fibroblasts (3T3) in tissue culture are electrically coupled to their interphase neighbors. Recordings from many such cells suggest that this coupling persists throughout the division cycle of the mitotic cell.     

Separation of type 2 toxins of Vibrio cholerae

Choleragenic toxin was separated from vascular permeability factor by ion-exchange chromatography of supernatants of dialyzed peptone cultures of Vibrio cholerae. The choleragenic toxin eluted from columns of QAE-Sephadex with low-ionicity systems is free of permeability factor activity. Further elution of these columns with 0.5M NaCl removes both the permeability factor and residual choleragenic toxin. When this latter material is chromatographed on columns of carboxymethyl-Sephadex, the permeability factor toxin is eluted by 0.02M phosphate buffer and is free of choleragenic activity. Therefore, choleragenic and permeability factor activities of the type 2 cholera toxins are different and can be separated by these procedures.     

冬枣黑疔病病原菌毒素致病机制研究

利用不同浓度的冬枣(Zizyphus jujuacv.Dongzao)黑疔病病原菌毒素提取液处理冬枣果皮组织,通过测定其对果皮组织细胞膜透性、丙二醛(MDA)含量变化、过氧化物酶(POD)、抗坏血酸过氧化物酶(APX)和过氧化氢酶(CAT)活性的影响,研究该毒素在致病机制中的作用。结果表明:经毒素处理后,果皮组织浸出液的相对电导率随毒素浓度增大而增加;细胞内丙二醛含量增加,高于对照,随毒素浓度的增大而升高;细胞内CAT活性在处理9 h内低于对照,随处理时间延长呈总体下降趋势,APX、POD活性在处理中期呈下降趋势,APX活性与对照差异不明显,POD活性在中后期与对照差异明显。     

Three-dimensional structure of cholera toxin penetrating a lipid membrane

Two-dimensional crystals of cholera toxin bound to receptors in a lipid membrane give diffraction extending to 15 A resolution. Three-dimensional structure determination reveals a ring of five B subunits on the membrane surface, with one-third of the A subunit occupying the center of the ring. The remaining mass of the A subunit appears to penetrate the hydrophobic interior of the membrane. Cleavage of a disulfide bond in the A subunit, which activates the toxin, causes a major conformational change, with the A subunit mostly exiting from the B ring.     

根腐菌毒素对菘蓝试管苗有丝分裂行为的影响

以窄叶菘蓝试管苗无性系为材料,采用常规压片技术及核型分析,研究了根腐菌毒素对细胞有丝分裂的影响。结果表明:在组织培养获得的植株中出现了四倍体细胞,在根腐毒素胁迫的无性植株中,除四倍体植株外,还有三体、四体等非整倍体细胞。此外,毒素胁迫还引起染色体断折、染色体桥等结构变异,对细胞分裂亦有一定的抑制作用。     

Amitotic neuroblastoma cells used for neural implants in monkeys

The potential utility of cultured neuroblastoma cells as donor tissue for neutral implants into the mammalian brain has been examined. Cells from a human neuroblastoma cell line, IMR-32, were labeled with [3H]thymidine and chemically rendered amitotic. These differentiated IMR-32 cells were grafted into the hippocampi of five adult African Green monkeys, and graft survival was evaluated for up to 270 days after transplantation. Autoradiographically labeled grafted cells were identified in four animals. Processes from grafted cells could be followed for distances of up to 150 micrometers into the host brain. No evidence for neoplastic growth of the transplant was found. Thus, grafted neuroblastoma cells can survive for prolonged periods in the primate brain and may serve as a practical source of donor tissue for neural implants.     

Human melanoma proteoglycan: expression in hybrids controlled by intrinsic and extrinsic signals

Human malignant melanoma cells express specific chondroitin sulfate proteoglycans (mel-CSPG) on the surface, both in vivo and in vitro. Melanocytes in normal skin show no detectable mel-CSPG but can be induced to express the antigen when cultured in the presence of cholera toxin and the tumor promoter 12-O-tetradecanoylphorbol-13-acetate. Most other cell types do not express mel-CSPG either in vivo or in vitro. A study was designed to examine regulatory signals controlling mel-CSPG expression. The gene encoding mel-CSPG was mapped to human chromosome 15, and this chromosome was introduced into rodent cells derived from distinct differentiation lineages. Three types of mel-CSPG--expressing hybrids were found: (i) hybrids derived from human melanomas; (ii) hybrids derived from human cells that do not express mel-CSPG; and (iii) hybrids derived from human cells expressing mel-CSPG that are antigen-negative but that are induced to express mel-CSPG when cultured on extracellular matrix instead of plastic surfaces. Thus, mel-CSPG expression can be controlled both through intrinsic signals, provided by the differentiation program of the rodent fusion partner, and through extrinsic signals, provided by specific cell-matrix interactions.     

Structural basis for the activation of cholera toxin by human ARF6-GTP

The Vibrio cholerae bacterium causes devastating diarrhea when it infects the human intestine. The key event is adenosine diphosphate (ADP)-ribosylation of the human signaling protein GSalpha, catalyzed by the cholera toxin A1 subunit (CTA1). This reaction is allosterically activated by human ADP-ribosylation factors (ARFs), a family of essential and ubiquitous G proteins. Crystal structures of a CTA1:ARF6-GTP (guanosine triphosphate) complex reveal that binding of the human activator elicits dramatic changes in CTA1 loop regions that allow nicotinamide adenine dinucleotide (NAD+) to bind to the active site. The extensive toxin:ARF-GTP interface surface mimics ARF-GTP recognition of normal cellular protein partners, which suggests that the toxin has evolved to exploit promiscuous binding properties of ARFs.     

Receptor-associated resistance to growth hormone-releasing factor in dwarf "little" mice

Anterior pituitaries from the dwarf mouse strain "little" did not release growth hormone or accumulate adenosine 3',5'-monophosphate (cyclic AMP) in response to human and rat growth hormone-releasing factor (GRF). Dibutyryl cyclic AMP, as well as the adenylate cyclase stimulators forskolin and cholera toxin, markedly stimulated growth hormone (GH) release. The basis of the GH deficiency in the little mouse may therefore be a defect in an early stage of GRF-stimulated GH release related either to receptor binding or to the function of the hormone-receptor complex.     

Tissue specificity of histone phosphorylation

The incorporation of (32)P-phosphate into histone fractions isolated from normal, hepatectomized, and Novikoff hepatoma-bearing rats was investigated. Varying degrees of phosphorylation were exhibited by different histone fractions. The phosphorylation of histones is tissue specific and appears to be correlated with metabolic cell functions, that is, it decreases with increasing mitotic activity of the cells.     

Direct evidence that endogenous GM1 ganglioside can mediate thymocyte proliferation

The B subunit of cholera toxin, which is multivalent and binds exclusively to a specific ganglioside, GM1, was mitogenic for rat thymocytes. When exposed to the B subunit, the cells proliferated, as measured by 3H-labeled thymidine incorporation. Mitogenesis depended on the direct interaction of the B subunit with GM1 on the surface of the cells. This demonstrates that endogenous plasma membrane gangliosides can mediate proliferation in lymphocytes.     

Sequence of the alpha subunit of photoreceptor G protein: homologies between transducin, ras, and elongation factors

A bovine retinal complementary DNA clone encoding the alpha subunit of transducin (T alpha) was isolated with the use of synthetic oligodeoxynucleotides as probes, and the complete nucleotide sequence of the insert was determined. THe predicted protein sequence of 354 amino acids includes the known sequences of four tryptic peptides and sequences adjacent to the residues that undergo adenosine diphosphate ribosylation by cholera toxin and pertussis toxin. On the basis of homologies to other proteins, such as the elongation factors of protein synthesis and the ras oncogene proteins, regions are identified that are predicted to be acylated and involved in guanine nucleotide binding and hydrolysis. Amino acid sequence similarity between T alpha and ras is confined to these regions of the molecules.     

镰刀菌毒素在小麦组织离体培养中的类激素活性

通过使用含镰刀菌粗毒素的诱导培养基,研究了镰刀菌毒素在小麦离体组织脱分化中的作用.发现禾谷镰刀菌粗毒素在小麦组织离体培养中具有关激素活性和毒害双重作用.毒素的类激素活性因所用外植体而异,在幼穗组织中表现较为显着,最适浓度下,3个供试品种的愈伤组织产量比对照均高出1倍.同时,镰刀菌毒素还能影响被诱导愈伤组织的继代和分化性能,降低其存活率、生长势和绿苗分化频率.镰刀菌毒素的类激素活性和毒害作用在不同抗性品种的实验中显示出毒素浓度上的差异,感抗品种之间具有明显的界限.     

Site-specific recombination between homologous chromosomes in Drosophila

The ability to mark a cell and its descendants genetically so that the resulting cell clone can be distinguished from neighboring cells facilitates studies in animal biology and development. A method of generating clones by inducing homologous mitotic recombination in Drosophila with a site-specific yeast recombinase is described. This method allows for frequent mosaicism after mitotic exchange is induced at predefined sites in the genome.     

Exploring and engineering the cell surface interface

Cells are inherently sensitive to local mesoscale, microscale, and nanoscale patterns of chemistry and topography. We review current approaches to control cell behavior through the nanoscale engineering of materials surfaces. Far-reaching implications are emerging for applications including medical implants, cell supports, and materials that can be used as instructive three-dimensional environments for tissue regeneration.     

转Bt基因抗虫棉Bt基因表达的时空动态

以转Bt基因抗虫棉 33B为材料 ,利用ELISA检测方法 ,通过对棉株不同发育时期和同一时期的不同组织或器官Bt晶体蛋白含量的测定 ,研究了转Bt基因抗虫棉Bt基因表达的时空变化。结果表明 ,随着棉株生育进程的推进和株体的老化 ,Bt晶体蛋白含量随着植株体内可溶性总蛋白含量的逐渐降低而降低 ,而Bt基因的表达强度从苗期到蕾期随着棉株营养生长的加快而呈上升趋势 ,至蕾期达到高峰 ,以后逐渐减弱。不同组织或器官Bt晶体蛋白的含量也有较大差异 ,表现在幼嫩组织或器官的含量较高 ,成熟组织或器官次之 ,衰老组织最低。这说明Bt基因表达强度的减弱和Bt晶体蛋白含量的降低是转Bt基因抗虫棉生育后期抗虫性降低的根本原因