Infinite average lifetime of an unstable bright state in the green fluorescent protein.

The time evolution of the fluorescence intensity emitted by well-defined ensembles of green fluorescent proteins has been studied by using a standard confocal microscope. In contrast with previous results obtained in single-molecule experiments, the photobleaching of the ensemble is well described by a model based on Lévy statistics. By assuming the presence of thermally activated barriers, this simple model allows us to obtain information about their height distribution.

Large phenotype jumps in biomolecular evolution.

By defining the phenotype of a biopolymer by its active three-dimensional shape, and its genotype by its primary sequence, we propose a model that predicts and characterizes the statistical distribution of a population of biopolymers with a specific phenotype that originated from a given genotypic sequence by a single mutational event. Depending on the ratio g(0) that characterizes the spread of potential energies of the mutated population with respect to temperature, three different statistical regimes have been identified. We suggest that biopolymers found in nature are in a critical regime with g(0) approximately 1-6, corresponding to a broad, but not too broad, phenotypic distribution resembling a truncated Lévy flight. Thus the biopolymer phenotype can be considerably modified in just a few mutations. The proposed model is in good agreement with the experimental distribution of activities determined for a population of single mutants of a group-I ribozyme.

Preferential sputtering induced stress domains and mesoscopic phase separation on CaF2(111)

We report the formation of novel mesoscopic two-dimensional bubble structures coexisting with nanometer-scale, two monolayer high, meandering islands of characteristic separation approximately 11 nm on CaF2(111) surfaces exposed to glancing incidence ion beam irradiation (4 degrees, 4.5 keV Ar+). The island and bubble structures can be explained, respectively, by nanoscale stress domain related local self-ordering of a single calcium adlayer on phase-separated F- (fluorine ion) and F-center (color center) terminated mesoscopic domains.

Effects of isoniazid on ultrastructure of Mycobacterium aurum and Mycobacterium tuberculosis and on production of secreted proteins.

Isoniazid (INH), one of the most effective antimycobacterial drugs, specifically inhibits, at an early stage of its action, the biosynthesis of mycolic acids, specific mycobacterial lipids which play a central role in the cell envelope architecture of mycobacteria. In the present study, the consequences of the action of INH on the cell morphology of Mycobacterium tuberculosis and Mycobacterium aurum were examined. Electron microscopy was used to observe bacilli which were previously treated with either subinhibitory concentrations of INH or the MIC of the drug, leading to a decrease of 20 to 35% (by weight) of their mycolic acid contents. The earlier effect of INH on the ultrastructure of mycobacteria, as revealed by negative staining of bacilli, was the alteration of the bacterial poles; this event was observed prior to the bacteriostatic action of the drug and was accompanied by a release of material from the poles into the extracellular medium. In a later stage of the drug's action, cell deformation occurred and more extracellular material was seen. The material released following the action of the drug on susceptible mycobacterial cells was identified as being almost exclusively composed of proteins. Labeling of amino acids with 35S prior to and during the action of INH on M. aurum and subsequent analysis of the labeled proteins led to the conclusion that they consisted of secreted proteins which were up to 20-fold oversecreted in the presence of the drug. Competitive enzyme-linked immunosorbent assay with the secreted 45/47-kDa antigen complex of M. tuberculosis demonstrated up to 20-fold oversecretion of these proteins. Taken together, the production of oversecreted proteins following the decrease of the cell envelope mycolate content by INH strongly suggests that mycolic acids may act as a barrier in the export of proteins secreted by mycobacteria.

Composition and phase behaviour of polar lipids isolated from Spirulina maxima cells grown in a perdeuterated medium.

The lipid composition of Spirulina maxima cells grown in a perdeuterated medium was determined by using nuclear magnetic resonance spectroscopy, fast atom-bombardment-mass spectrometry, gas chromatography-mass spectrometry as well as conventional chemical methods. The extent of deuteration was determined by mass spectrometry and was superior to 97.5%. The major lipids identified in the strain were: non-polar lipids (9%), monogalactosyldiacylglycerol (5%), digalactosyldiacylglycerol (22%), phosphatidylglycerol (31%), sulfoquinovosyldiacylglycerol (32%), phosphatidylinositol (traces). The major fatty acids were 16:0 (80%) and 18:1 (15%). These results demonstrate that the adaptation of the cells to D2O did not imply a profound modification of the lipid composition. The perdeuterated polar lipid mixture dispersed into an excess of water organises spontaneously in a lamellar phase as seen by 31P and deuterium solid state NMR and can therefore be used to prepare perdeuterated model membranes with a well defined composition. Liposomes made using these lipids have a gel to liquid-crystalline phase transition in the range 15-27 degrees C and are in a fluid L alpha phase above this temperature.

Identification of the surface-exposed lipids on the cell envelopes of Mycobacterium tuberculosis and other mycobacterial species.

The surface-exposed lipids of Mycobacterium tuberculosis, Mycobacterium avium, Mycobacterium kansasii, Mycobacterium gastri, Mycobacterium smegmatis, and Mycobacterium aurum were isolated by gentle mechanical treatment of cells with glass beads. Analysis of the exposed lipids demonstrated a selective location of classes of ubiquitous lipids on the surfaces of mycobacteria. While phosphatidylethanolamine and phosphatidylinositol mannosides were exposed in all the species examined, dimycoloyl trehalose ("cord factor") was identified in the surface components of M. aurum only. Furthermore, monomycoloyl trehaloses and triacylglycerides were identified in the surface-exposed lipids of M. avium and M. smegmatis but not in those of the other mycobacterial species examined. The species- and type-species specific lipids were present on the mycobacterial cell surface: phenolic glycolipids, dimycocerosates of phthiocerols, and lipooligosaccharides were identified in the surface-exposed materials of M. tuberculosis (Canetti), M. kansasii, and M. gastri, whereas glycopeptidolipids were identified in the outermost lipid constituents of M. avium and M. smegmatis. This difference in the surface exposure of lipids of various mycobacterial species may reflect differences in their cell envelope organizations. Brief treatments of M. tuberculosis with Tween 80 prior to the use of glass beads led to erosion of regions of the capsule to expose gradually both cord factor and other lipids on the cell surface of the tubercle bacillus, demonstrating that the latter lipids are buried more deeply in the cell envelope and leading to the proposal of a scheme for the location of the capsular lipids of the tubercle bacillus.