Membrane Targeting and GTPase Activity of Rab7 Are Required for Its Ubiquitination by RNF167

Rab7 is a GTPase that controls late endosome and lysosome trafficking. Recent studies have demonstrated that Rab7 is ubiquitinated, a post-translational modification mediated by an enzymatic cascade. To date, only one ubiquitin E3 ligase and one deubiquitinase have been identified in regulating Rab7 ubiquitination. Here, we report that RNF167, a transmembrane endolysosomal ubiquitin ligase, can ubiquitinate Rab7. Using immunoprecipitation and in vitro ubiquitination assays, we demonstrate that Rab7 is a direct substrate of RNF167. Subcellular fractionation indicates that RNF167 activity maintains Rab7′s membrane localization. Epifluorescence microscopy in HeLa cells shows that Rab7-positive vesicles are larger under conditions enabling Rab7 ubiquitination by RNF167. Characterization of its ubiquitination reveals that Rab7 must be in its GTP-bound active form for membrane anchoring and, thus, accessible for RNF167-mediated ubiquitin attachment. Cellular distribution analyses of lysosome marker Lamp1 show that vesicle positioning is independent of Rab7 and RNF167 expression and that Rab7 endosomal localization is not affected by RNF167 knockdown. However, both Rab7 and RNF167 depletion affect each other’s lysosomal localization. Finally, this study demonstrates that the RNF167-mediated ubiquitination of Rab7 GTPase is impaired by variants of Charcot–Marie–Tooth Type 2B disease. This study identified RNF167 as a new ubiquitin ligase for Rab7 while expanding our knowledge of the mechanisms underlying the ubiquitination of Rab7.     

Pure Chitosan Biomedical Textile Fibers from Mixtures of Low- and High-Molecular Weight Bidisperse Polymer Solutions: Processing and Understanding of Microstructure–Mechanical Properties’ Relationship

Natural polymers, as extracted from biomass, may exhibit large macromolecular polydispersity. We investigated the impact of low molar mass chitosan (LMW, DP_w~115) on the properties of chitosan fibers obtained by wet spinning of chitosan solutions with bimodal distributions of molar masses. The fiber crystallinity index (CrI) was assessed by synchrotron X-ray diffraction and the mechanical properties were obtained by uniaxial tensile tests. The LMW chitosan showed to slightly increase the crystallinity index in films which were initially processed from the bimodal molar mass chitosan solutions, as a result of increased molecular mobility and possible crystal nucleating effects. Nevertheless, the CrI remained almost constant or slightly decreased in stretched fibers at increasing content of LMW chitosan in the bidisperse chitosan collodion. The ultimate mechanical properties of fibers were altered by the addition of LMW chitosan as a result of a decrease of entanglement density and chain orientation in the solid state. An increase of crystallinity might not be expected from LMW chitosan with a still relatively high degree of polymerization (DPw ≥ 115). Instead, different nucleation agents—either smaller molecules or nanoparticles—should be used to improve the mechanical properties of chitosan fibers for textile applications.     

Effects of photoperiod and pair-feeding on lactation of cows fed corn or barley grain in total mixed rations

Effects of photoperiod and type of grain were assessed using 47 cows (wk 1; 59.5 d of lactation) exposed daily to 16 h of light (long day) and fed randomly one of two diets containing an equal amount of cracked corn or rolled barley. During wk 5, cows were assigned within each diet to long day and ad libitum feeding, short day (8 h of light: 2 h of dark: 2 h of light: 12 h of dark) and ad libitum feeding, or long day and ad libitum feeding (wk 5 to 8) followed by pair feeding with short day cows between wk 9 and 16. Milk composition was not affected by treatments. Between wk 9 and 16, long day cows fed for ad libitum consumption or pair fed produced 5 to 11% more milk than cows exposed to short day; feed intake of long day cows fed for ad libitum consumption was greater than short short day and long day cows pair fed cows. Milk yield and total feed intake were not affected by type of grain in the diet. In conclusion, long day photoperiod increased milk yield and feed intake.     

The PANOPTIC Camera: A Plenoptic Sensor with Real-Time Omnidirectional Capability

A new biologically-inspired vision sensor made of one hundred “eyes” is presented, which is suitable for real-time acquisition and processing of 3-D image sequences. This device, named the Panoptic camera, consists of a layered arrangement of approximately 100 classical CMOS imagers, distributed over a hemisphere of 13 cm in diameter. The Panoptic camera is a polydioptric system where all imagers have their own vision of the world, each with a distinct focal point, which is a specific feature of the Panoptic system. This enables 3-D information recording such as omnidirectional stereoscopy or depth estimation, applying specific signal processing. The algorithms dictating the image reconstruction of an omnidirectional observer located at any point inside the hemisphere are presented. A hardware architecture which has the capability of handling these algorithms, and the flexibility to support additional image processing in real time, has been developed as a two-layer system based on FPGAs. The detail of the hardware architecture, its internal blocks, the mapping of the algorithms onto the latter elements, and the device calibration procedure are presented, along with imaging results.     

Use of signal processing imaging for the study of a 3D package in harsh environment

Assembly technologies follow the reduction in size of circuit board components and correlatively the number of active layers increases significantly. Therefore, during a conventional acoustic analysis, the obtained image becomes more and more complicated to analyze. In a previous paper, we focused our research on a method for a suitable visualization of the timescale of an acoustic signal (CWT). In this paper we present the use of the non-destructive control for the study of a 3D package submitted to accelerated ageing tests using harsh conditions.     

Quantitative luminescence mapping of Cu(In,Ga)Se2 thin‐film solar cells

We investigate photoluminescence and electroluminescence (PL and EL) emission images from Cu(In,Ga)Se₂‐based solar cells by means of a Hyperspectral Imager. Using the generalized Planck's law, maps of the effective quasi‐Fermi level splitting Δμ_eff in absolute values are obtained. A good agreement is found between the spatially averaged splitting in PL and the global open‐circuit voltage. However, from a local carrier transport discussion, we conclude that the equality does not hold locally. The spatial variations are rather attributed to local depth variations of the quasi‐Fermi level splitting due to material properties spatial fluctuations. By comparing PL and EL emissions, we discuss qualitatively the local effective lifetimes and collection efficiencies. Copyright © 2014 John Wiley & Sons, Ltd.     

High pass filter with above IC integrated SrTiO3 high K MIM capacitors

This paper describes realization and characterization of SrTiO₃ (STO) high K MIM capacitors above BiCMOS integrated circuit (IC). These capacitances are connected to IC and are used as coupling capacitors in order to realize a high pass filter. Surface capacitance achieved is 10 nF/mm² with capacitance value of 1.2 nF. The process for STO MIM fabrication does not exceed 400 °C, which is compatible with interconnections. Typical K and dielectric losses values obtained are, respectively 110% and 2%. Yield obtained reaches 83% for capacitors. A functional high pass filter using these STO capacitors was realized in this study. It exhibits a cut-off frequency at 6.5 kHz and a constant gain at higher frequencies of −1.3 dB.