The phylogenic expression of plasmolipin in the vertebrate nervous system

Plasmolipin is a plasma membrane proteolipid is a major myelin membrane component (Cochary et al., 1990). In this study we report the phylogenic expression of plasmolipin in the vertebrate nervous system. Using Western blot analysis with polyclonal antibodies, we have analyzed membrane fractions, including myelin, from elasmobranchs, teleosts, amphibians, reptiles, birds and mammals. On the basis of immune detection, plasmolipin appears to be restricted to the mammalian nervous system. Comparison of the central and peripheral nervous systems of mammals showed only minor differences in the level of plasmolipin in these two regions. Within mammals, little quantitative differences were observed when rat, human and bovine membrane fractions were compared. The late evolutionary expression of plasmolipin which results in its restriction to mammals makes it unique among the (major) myelin proteins. The potential physiologic significance of these data are discussed.Abbreviations EDTA Ethylene diamine N.,NN tetracetic acid - EGTA Ethylene glycol bis-(B-Aminoethyl Ether) N,,NN tetracetic acid - MES ([N-Morpholino] ethane sulfonic acid) DCCD, N, Dicyclohexyl carbodiimide     

Aerial Dispersal and Epiphytic Survival of Pseudomonas syringae during a Pretest for the Release of Genetically Engineered Strains into the Environment

Prospective experimental field evaluation of genetically engineered microorganisms, such as microbial pest control agents, raises issues of how to properly ascertain their fate and survival in the environment. Field trials with recombinant organisms must reflect requirements for sampling and monitoring. Field trials were conducted at Tulelake, Calif., to monitor the numbers of viable cells of a nonrecombinant strain of Pseudomonas syringae that entered the atmosphere and landed on plants and soil during and after an aerosol spray application. An exponential decrease in numbers of viable cells deposited at increasing distances from three sprayed plots was observed. The relative rate of survival of cells sprayed directly on plants was more than 10 times higher than that of cells dispersed through the air to similar adjacent plants. Results are being used to gain experience with the characteristics of a release site that influence containment or dispersal and to develop appropriate sampling methodologies for evaluating survival and dispersal characteristics of genetically engineered bacteria released into the environment. The ability to make predictions about microbial dispersal and survival will reduce the uncertainties associated with environmental releases of recombinant organisms.     

Extracellular Fluid Proteins of Goldfish Brain: Evidence for the Presence of Proteases and Esterases

Preparations of enriched fractions of extracellular fluid (ECF) proteins from goldfish brain were found to contain protease(s) and esterase(s). The N-substituted furanacryloyl (FA) peptides FA-Phe-Gly-Gly and FA-Phe-OMe were used as model substrates for determining protease and esterase activity, respectively, in a spectrophotometric assay. Studies of the profile of substrate specificity and identification of the types of compounds that were effective as inhibitors showed that these ECF enzymes have some distinctive properties. GSH, but not GSSG, and EDTA inhibited the protease(s) without influencing the esterase(s), whereas L-1-tosylamide-2-phenylethylchloromethyl ketone blocked both protease and esterase activities of ECF. Most of the protease and esterase properties of ECF could be bound to concanavalin A-Sepharose affinity chromatographic columns in association with ependymin--a brain extracellular protein. These observations indicate that ECF may contain a metalloprotease(s) and raise the possibility that the ependymins might be a substrate for these ECF enzymes.     

Purification of human plasma retinol-binding protein by hydrophobic interaction chromatography

Human plasma retinol-binding protein has been purified to homogeneity by a simple method that requires an ammonium sulfate fractionation, a hydrophobic interaction chromatography on phenyl-Sepharose, which dissociates the complex between retinol-binding protein and its carrier, transthyretin, and a gel filtration on Sephadex G-50. The yield of pure protein is comparable or higher than that obtained with the more complex procedures previously reported.     

Procedure for isolation of gangliosides in high yield and purity: simultaneous isolation of neutral glycosphingolipids

While several methods for ganglioside extraction and isolation have been described, relatively little attention has been given to the effectiveness of separation from peptides, phospholipids, and various low-molecular-weight contaminants. A procedure is described for isolation of gangliosides in high purity and good yield from 1- to 400-mg samples (wet wt). A key step was mild acidification following homogenization, designed to dissociate gangliosides from lipophilic peptides which coextracted into organic solvents. This has proved particularly helpful for myelin and myelin-containing tissues (e.g., white matter, nerve) whose proteins have presented special problems in ganglioside isolation. In this study isolation was effected by consecutive chromatographies on Sephadex LH-20, DEAE-Sephadex, and silica gel following the initial acidification. The method applied to bovine white matter gave tissue concentrations (calculated from yields and radiolabeled tracer recoveries) that were similar to those obtained with three previously described procedures; however, peptide contaminants were an order of magnitude lower. Removal of low-molecular-weight contaminants, including nucleotide sugars, was virtually complete. In addition to ganglioside isolation the method can be used to obtain neutral glycosphingolipids as well. It is believed to have broad applicability to a diversity of tissues.     

Influence of aerial dispersal on persistence and spread of pesticide-resistant Metaseiulus occidentalis in California almond orchards

Aerial dispersal of the phytoseiid Metaseiulus occidentalis (Nesbitt) was evaluated as a component in managing pesticide-resistant populations established in California almond orchards. Peak dispersal occurred in late July and early August during 1982 and 1983. Most predators (and spider mites) left the orchards on the prevailing winds from the northwest. Within the orchard, the prevailing winds had less influence, and dispersal was usually random. Both spider mites and predators dispersed randomly with regard to height from the almond trees, but data obtained during one 24-h interval suggest they do not disperse randomly throughout the day. Most aerial movements occurred between 16–22 h when relative humidity and wind speeds increased and temperatures decreased. Spider mites and predators were trapped on panels located 200 m from the orchard. A survey of carbaryl resistance levels in M. occidentalis collected from almond orchards surrounding the release sites indicates that carbaryl-resistant M. occidentalis dispersed at least 800 m between 1981–83. However, growers wishing to use the resistant strains should release them in their orchards as natural dispersal appears to be too slow. Migration of native M. occidentalis into the release sites appeared to be sufficiently rare that dilution of carbaryl-resistant populations was minimal during a 2–4 year period.

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Résumé La dispersion aérienne du phytoseïdae, M. occidentalis (Nesbitt), a été estimée comme élément de la lutte contre les populations résistantes aux insecticides établies dans les vergers de Californie. La dispersion maximale s'est produite fin juillet et début a oût en 1982 et 1983. La plupart des prédateurs (et des acariens) quittent les vergers avec les vents dominants du nordouest. Dans le verger, les vents dominants sont moins importants et la dispersion est généralement au hasard. Tant les acariens que les prédateurs se dispersaient au hasard par rapport à la taille des amandiers, mais les relevés sur 24 heures laissent supposer qu'il n'y a pas une distribution aléatoire pendant la journée. La plupart des mouvements aériens se produisirent entre 16 et 22 heures quand HR et vitesse du vent augmentaient et température diminuait. Les acariens et prédateurs ont été piégés sur des panneaux à 200 m du verger.

     

Monoclonal antibodies against neoantigens of the terminal C5b-9 complex of human complement

Assembly of the terminal C5b-C9 complement components into the cytolytic C5b-9 complex is accompanied by exposure of characteristic neoantigens on the macromolecule. We report the production and characterization of mouse monoclonal antibodies to C9-dependent neoantigens of human C5b-9. Binding-inhibition assays with EDTA-human plasma and micro-ELISA assays with purified C9 showed that the antibodies did not react with native complement components and thus confirmed the specificity of the antibodies for the neoantigens. The monoclonal antibodies did, however, cross-react with cytolyticaIly inactive, fluid-phase C5b-9 complexes, Thus, expression of the neoantigenic determinants was not dependent on the formation of high molecular weight C9 polymers with the complex, since these are absent in fluid-phase C5b-9. Radioiodinated antibodies could be utilized in immunoradiometric assays for the detection and quantitation of C5b-9 on cell membranes. Cross-reactivities of the antibodies with C9-dependent neoantigens of several other animal species were examined and antibody clones cross-reacting with rabbit (clones 3BI, 3Dg, and 2F3), sheep (clones 3Dg and 2F3) and guinea-pig (clone 3D8) neoantigens were identified . Three of four tested clones (3D8, 2F3, IA12) precipitated C5b-9 complexes in double-diffusion assays, probably due to their interaction with multiple and repeating C9-epitopes on the terminal complexes. The monoclonal antibodies will be of value for definitive identification and quantitation of C5b-9 on cell membranes and in tissues, and for establishing immunoassays for detection and quantitation of terminal fluid-phase C5b-9 complexes in plasma.     

14C-Metabolism in cultured cotyledon explants of radiata pine

Excised cotyledons of radiata pine ( Pinus radiata D. Don), cultured under shootforming (plus cytokinin) and elongating (minus cytokinin) conditions, were incubated in ¹⁴C-glucose, ¹⁴C-acetate or ¹⁴C-bicarbonate at different stages of growth and differentiation. ¹⁴CO₂ was produced when the cotyledons were fed ¹⁴C-glucose and ¹⁴C-acetate (no measurement was made for ¹⁴C-bicarbonate feeding). Label from these precursors was incorporated into ethanol-soluble and -insoluble fractions. The largest percentage of radioactivity was associated with the ethanol-soluble portion, which was further fractionated into lipids, amino acids, organic acids and sugars. The amount of label and the pattern of labelling associated with each of the above classes of metabolites varied with time in culture and morphogenetic behaviour of the cotyledons. In general, there was a tendency towards a high rate of incorporation of label in elongating cotyledons during the period of rapid elongation. On the other hand, a high rate of incorporation of label in shoot-forming cotyledons coincided with the period of meristematic tissue formation. The data obtained support the hypothesis that organized development in vitro involves a shift in metabolism, which precedes and is coincident with the initiation of the process.