Entry of Human Papillomavirus Type 16 by Actin-Dependent, Clathrin- and Lipid Raft-Independent Endocytosis

Infectious endocytosis of incoming human papillomavirus type 16 (HPV-16), the main etiological agent of cervical cancer, is poorly characterized in terms of cellular requirements and pathways. Conflicting reports attribute HPV-16 entry to clathrin-dependent and -independent mechanisms. To comprehensively describe the cell biological features of HPV-16 entry into human epithelial cells, we compared HPV-16 pseudovirion (PsV) infection in the context of cell perturbations (drug inhibition, siRNA silencing, overexpression of dominant mutants) to five other viruses (influenza A virus, Semliki Forest virus, simian virus 40, vesicular stomatitis virus, and vaccinia virus) with defined endocytic requirements. Our analysis included infection data, i.e. GFP expression after plasmid delivery by HPV-16 PsV, and endocytosis assays in combination with electron, immunofluorescence, and video microscopy. The results indicated that HPV-16 entry into HeLa and HaCaT cells was clathrin-, caveolin-, cholesterol- and dynamin-independent. The virus made use of a potentially novel ligand-induced endocytic pathway related to macropinocytosis. This pathway was distinct from classical macropinocytosis in regards to vesicle size, cholesterol-sensitivity, and GTPase requirements, but similar in respect to the need for tyrosine kinase signaling, actin dynamics, Na+/H+ exchangers, PAK-1 and PKC. After internalization the virus was transported to late endosomes and/or endolysosomes, and activated through exposure to low pH

ZAP: Um Algorítmo de Atribuição Distribuída de Canais para Mitigação de Interferências em Redes com Rádio Cognitivo

International audienceThis paper presents ZAP, a protocol for distributed assignment of frequency channels to links in cognitive radio networks. These radios are capable of identifying under-utilized licensed bands of the spectrum, allowing their reuse without interfering with primary users. The frequency assignment must be simple, incur acceptable communication overhead, provide timely response and be adaptive to accommodate constant changes in the network. Another challenge is the optimization of network capacity through interference minimization. Unlike the related proposals in the literature, ZAP responds to this challenge with a distributed approach based only on local (neighborhood) knowledge, while significantly reducing computational cost and number of messages required for channel assignment. Simulations confirm the good quality of ZAP in terms of (1) performance compromise between different metrics and (2) fast solution achievement regardless of network size

Effect of Inflow Conditioning for Dry Powder Inhalers

The transport of pharmaceutical dry powder inside an optically accessible inhaler-like device is studied using both macro- and microscopic high-speed imaging. The investigation aims to systematically study the effect of inflow modifications on the dispersion characteristics of agglomerates inside a dry powder inhaler (DPI) geometry. An inhaler device was designed with geometrical features akin to commercial inhalers used in the current market and research oriented inhalers such as the Twincer®: two offset inlet channels (one with a powder pocket), a clockwise swirling chamber and a single outlet channel. At the device outlet, a vacuum pump was fitted with an actuator and calibrated to achieve a steady state inhalation with a peak flowrate of 85 and 125 L/min. Airflow conditions at the intake of the device were strategically perturbed in order to induce powder fluidisation and dispersion using turbulence grids and through physically obstructing channel streams in order to achieve changes in flow behaviour (e.g., flow separation). Complete fluidisation of the powder bed was observed with image processing enabling statistics on de-agglomerated fragment size and velocity. A range of behaviour was noted including local turbulence through introduction of a grid, bimodal fragment size behaviour for cohesive mannitol powder, as well as introduction of low velocity zones in the device through flow splitting. The geometry enables simple systematic study of inflow conditions into a DPI-like device with the data being useful for study of a given powder formulation (mannitol) and validation of computational models