Intracellular localization of two betaine lipids by cell fractionation and immunomicroscopy

The cellular localization of the betaine lipids diacylglyceryl-N,N,N-trimethylhomoserine (DGTS) and diacylglycerylhydroxymethyl-N,N,N-trimethyl-beta-alanine (DGTA) was investigated by a) chemical analysis of subcellular fractions and b) immunochemical methods using specific antisera and either fluorescence microscopy or electron microscopy for detection of the label. A homogenate of Lycopodium annotinum (Pteridophyta) was fractionated by differential and density gradient centrifugation. The particulate fractions obtained were analyzed for chlorophyll, cyt c oxidase, NADH-cyt c reductase and DGTS. Non-plastidial fractions were enriched in DGTS and only minor amounts of this lipid could be attributed to chloroplasts. Anti-DGTS and anti-DGTA sera were produced by immunization of rabbits. The monospecificity of the antisera was examined with cells of Chlamydomonas reinhardtii (Chlorophyceae) containing DGTS, Pavlova lutheri (Haptophyceae) containing DGTA and Ochromonas danica (Chrysophyceae) containing both DGTS and DGTA. Euglena gracilis which is free of betaine lipids, was used as a control. For the test, a FITC-coupled goat anti-rabbit antibody was used and detected by fluorescence microscopy. Thin sections of Ochromonas and Pavlova were incubated first with the anti-lipid sera and subsequently with a gold-coupled anti-rabbit serum and then examined in the electron microscope. With Ochromonas, anti-DGTS as well as anti-DGTA sera gave an accumulation of gold label in the cytoplasmic space but not in the chloroplasts. Similar results were obtained with Pavlova using anti-DGTA serum. These results describe for the first time the cytochemical localization of DGTS and DGTA strongly suggesting both these lipids to be associated mainly with non-plastidial structures.     

Zur Analyse von Petrolsulfonaten

Analysis of Petroleum Sulfonates The application of column chromatography using silica gel and ion-exchange resins has been studied for the separation of components of petroleum sulfonates present in technical products. The customary standard procedures based on extraction have been replaced by column chromatography, since the latter is more reliable and rapid. The various petroleum sulfonates and other products of sulfonation are differentiated by spectroscopic methods.     

How to Build an Ideal Cipher: The Indifferentiability of the Feistel Construction

This paper provides the first provably secure construction of an invertible random permutation (and of an ideal cipher) from a public random function that can be evaluated by all parties in the system, including the adversary. The associated security goal was formalized via the notion of indifferentiability by Maurer et al. (TCC 2004). The problem is the natural extension of that of building (invertible) random permutations from (private) random functions, first solved by Luby and Rackoff (SIAM J Comput 17(2):373–386, 1988) via the four-round Feistel construction. As our main result, we prove that the Feistel construction with fourteen rounds is indifferentiable from an invertible random permutation. We also provide a new lower bound showing that five rounds are not sufficient to achieve indifferentiability. A major corollary of our result is the equivalence (in a well-defined sense) of the random oracle model and the ideal cipher model.     

Overproduction of the Bradyrhizobium japonicum c-type cytochrome subunits of the cbb3 oxidase in Escherichia coli

We report on a system to improve expression of mature c-type cytochromes in Escherichia coli. It is based on the use of plasmid pEC86 that expresses the E. coli cytochrome c maturation genes ccmABCDEFGH constitutively, whereby the production of both endogenous and foreign c-type cytochromes was increased substantially. The periplasmic soluble domains of the c-type cytochrome subunits FixO and FixP of the Bradyrhizobium japonicum cbb3 oxidase could be expressed in E. coli only when pEC86 was provided in a degP-deficient strain. This shows that a stimulation of heme attachment by the Ccm maturase system combined with the diminished proteolytic activity in the periplasm can increase c-type cytochrome yields.     

Monitoring the WCl6/(CH3)4Sn initiated polymerization of substituted acetylenes by 1H-NMR spectroscopy

Summary A study of the WCl₆/(CH₃)₄Sn/CD₂Cl₂ initiated polymerization of otrimethylsilylphenylacetylene and 2-hexyne was completed. This study suggests that generated carbenes initiate the polymerization of mono and disubstituted acetylenes. Evidence was provided which clearly shows the formation ofthe initiating [W]=CH₂ carbene complex and its immediate disappearance upon addition of both the 2-hexyne and o-trimethylsilylphenylacetylene. Under the reaction conditions used in the present study, no spectroscopic evidence of an initiating or propagating chain end or metallocyclobutene intermediate could be provided.     

Methacrylate-capped fluoro side chain siloxanes: Synthesis,characterization, and their use in the design of oxygen-permeable hydrogels

Methacrylate end-capped 3-(2,2,3,3,-tetrafluoropropoxy)propyl, 3-(2,2,3,3,4,4,5,5-octafluoropentoxy)propyl, and 3-(2,2,3,3,4,4,5,5,6,6,7,7-dodecafluorotridecoxy)propyl side chain siloxanes were evaluated for potential use in hydrogels for contact lens application. The preparation of the methacrylate end-capped perfluoro side chain siloxanes was accomplished in two relatively simple synthetic steps. The first step consisted of the acid-catalyzed co-ring opening polymerization of octamethylcyclotetrasiloxane, tetramethylcyclotetrasiloxane, and 1,3-bis-methacryloxybutyltetramethyldisiloxane, followed by a platinum-catalyzed hydrosilation with the tetrafluoro-, octafluoro-, and dodecafluoro-substituted allylic ethers. The incorporation of the terminal [—CF₂—H] functionality had a dramatic effect on the increased solubility of the fluoro siloxane with hydrophilic monomers. Radical bulk polymerization of the methacrylate-capped fluoro side chain siloxanes with hydrophilic monomers, such as dimethylacrylamide and N-vinyl pyrrolidinone, resulted in transparent hydrogels possessing a wide range of water contents, high oxygen permeability, and a low modulus of elasticity. © 1997 John Wiley & Sons, Inc. J Appl Polym Sci 65:1081–1089, 1997     

Yeast Los1p has properties of an exportin-like nucleocytoplasmic transport factor for tRNA

Saccharomyces cerevisiae Los1p, which is genetically linked to the nuclear pore protein Nsp1p and several tRNA biogenesis factors, was recently grouped into the family of importin/karyopherin-beta-like proteins on the basis of its sequence similarity. In a two-hybrid screen, we identified Nup2p as a nucleoporin interacting with Los1p. Subsequent purification of Los1p from yeast demonstrates its physical association not only with Nup2p but also with Nsp1p. By the use of the Gsp1p-G21V mutant, Los1p was shown to preferentially bind to the GTP-bound form of yeast Ran. Furthermore, overexpression of full-length or N-terminally truncated Los1p was shown to have dominant-negative effects on cell growth and different nuclear export pathways. Finally, Los1p could interact with Gsp1p-GTP, but only in the presence of tRNA, as revealed in an indirect in vitro binding assay. These data confirm the homology between Los1p and the recently identified human exportin for tRNA and reinforce the possibility of a role for Los1p in nuclear export of tRNA in yeast.