ESCRT-III activation by parallel action of ESCRT-I/II and ESCRT-0/Bro1 during MVB biogenesis

The endosomal sorting complexes required for transport (ESCRT) pathway facilitates multiple fundamental membrane remodeling events. Previously, we determined X-ray crystal structures of ESCRT-III subunit Snf7, the yeast CHMP4 ortholog, in its active and polymeric state (<xref ref-type="bibr" rid="bib20">Tang et al., 2015</xref>). However, how ESCRT-III activation is coordinated by the upstream ESCRT components at endosomes remains unclear. Here, we provide a molecular explanation for the functional divergence of structurally similar ESCRT-III subunits. We characterize novel mutations in ESCRT-III Snf7 that trigger activation, and identify a novel role of Bro1, the yeast ALIX ortholog, in Snf7 assembly. We show that upstream ESCRTs regulate Snf7 activation at both its N-terminal core domain and the C-terminus α6 helix through two parallel ubiquitin-dependent pathways: the ESCRT-I-ESCRT-II-Vps20 pathway and the ESCRT-0-Bro1 pathway. We therefore provide an enhanced understanding for the activation of the spatially unique ESCRT-III-mediated membrane remodeling.

Luminescence of ZnO Nanocrystals in C21H38NCl and C23H14O11Na2 Matrixes

We report the data concerning investigations of photoluminescence spectra of ZnO nanoparticles in the matrixes of liquid crystal materials performed at T=290 K within a visible region. The mentuhflecioned spectra were obtained at polymeric state of Cetylpyridinium Chloride and Disodium Cromoglycate materials. The model of the radiation centers and mechanisms is discussed.

High-Pressure Studies of the Rotor-Stator Compound C60-Cubane

Insertion of cubane (C8H8) into the octahedral voids of the C60 lattice leads to the formation of an interesting rotor-stator compound which can be converted into a C60 co-polymer by heating. We have treated a number of C60·C8H8 samples for up to 3 h each in the range 380-875 K under pressures up to 2 GPa. The resulting materials were investigated by Raman spectroscopy and X-ray diffraction. Depending on treatment conditions, at least five different structural phases can be found. In addition to the four structural phases observed at atmospheric pressure and different temperatures we find that a new polymeric state is created at pressures above 1 GPa, and we tentatively identify its structure as pseudo-orthorhombic. The cubic-orthorhombic phase transition line is found to have a slope of 295 K GPa-1, much larger than the slope of the fcc-sc line in pure C60.

Paraformaldehyde fixation of neutrophils for immunolabeling of granule antigens in cryoultrasections.

Paraformaldehyde (PFA) fixation was optimized to facilitate the immobilization and labeling of multiple granule antigens, using short fixation regimens and cryoultramicrotomy of unembedded neutrophils (PMNs). In the optimal protocol, extraction of azurophil granule antigens (especially of the abundant elastase) was obviated by manipulating the polymeric state of PFA, and hence its rate of cross-linking, by altering its concentration and pH in a multistep process. Primary fixation conditions used (4% PFA, pH 8.0, 5 min) favor fixative penetration and rapid cross-linking. Stable cross-linking of the antigen was achieved in a secondary fixation step using conditions that favor larger, more cross-linking polymeric forms of PFA (8% PFA, pH 7.2, 15 min). Immobilization of granule antigens was enhanced by flotation of cut sections on fixative (8% PFA, pH 8.0) before labeling and by using post-labeling fixation with 1% glutaraldehyde. The optimized protocol facilitated immobilization and immunolabeling of elastase, myeloperoxidase, lactoferrin, and cathepsin D in highly hydrated, unembedded PMNs.

Mechanical Properties of Pyrolytic "SiOCN" Thin Coatings

ABSTRACTPyrolysis of polymer precursors (polysilazane) is a technologically and economically interesting way to produce thin ceramic coatings. However, many cracks appear and decohesion occurs during pyrolysis when the ceramic coatings (SiOCN) are thicker than 0.5 micrometers. In order to understand these cracking phenomena, the coatings are mechanically characterized by nanoindentation at different stages of the pyrolysis heat treatment.During pyrolysis, the cracking temperature is detected by in-situ optical observation. The thickness of the coatings varies during pyrolysis from 3 micrometers at the polymeric state to 1 micrometer at the ceramic state. The coatings' properties, hardness and Young's modulus are evaluated after heat treatment, taking into account the substrate's influence. A large variation of these properties occurs at the cracking temperature. Both the hardness and the Young's modulus are multiplied by a factor of 10. By analysing these results, we show that cracking is correlated with the evolution of the coatings' mechanical properties during the transformation.