GOLPH3 Regulates EGFR in T98G Glioblastoma Cells by Modulating Its Glycosylation and Ubiquitylation

Protein trafficking is altered when normal cells acquire a tumor phenotype. A key subcellular compartment in regulating protein trafficking is the Golgi apparatus, but its role in carcinogenesis is still not well defined. Golgi phosphoprotein 3 (GOLPH3), a peripheral membrane protein mostly localized at the trans-Golgi network, is overexpressed in several tumor types including glioblastoma multiforme (GBM), the most lethal primary brain tumor. Moreover, GOLPH3 is currently considered an oncoprotein, however its precise function in GBM is not fully understood. Here, we analyzed in T98G cells of GBM, which express high levels of epidermal growth factor receptor (EGFR), the effect of stable RNAi-mediated knockdown of GOLPH3. We found that silencing GOLPH3 caused a significant reduction in the proliferation of T98G cells and an unexpected increase in total EGFR levels, even at the cell surface, which was however less prone to ligand-induced autophosphorylation. Furthermore, silencing GOLPH3 decreased EGFR sialylation and fucosylation, which correlated with delayed ligand-induced EGFR downregulation and its accumulation at endo-lysosomal compartments. Finally, we found that EGF failed at promoting EGFR ubiquitylation when the levels of GOLPH3 were reduced. Altogether, our results show that GOLPH3 in T98G cells regulates the endocytic trafficking and activation of EGFR likely by affecting its extent of glycosylation and ubiquitylation.     

Evaluation of hydrogen metabolism by Escherichia coli strains possessing only a single hydrogenase in the genome

The four hydrogenase isozymes; hydrogenase 1 (Hyd-1), hydrogenase 2 (Hyd-2), hydrogenase 3 (Hyd-3) and hydrogenase 4 (Hyd-4) of Escherichia coli have been reported for their crucial functions in the hydrogen metabolism; however, their distinctive roles could not be completely understood. In this study, four ideal hydrogenase operon mutants, Δhyb hyc hyf, Δhya hyc hyf, Δhya hyb hyf, and Δhya hyb hyc, in which only a single hydrogenase is intact in the genome, were constructed as well as one quadruple mutant (Δhya hyb hyc hyf) that all four hydrogenase operons were deleted. First, single operon mutants and single-gene mutants for each hydrogenase showed different hydrogen productivity and growth in the anaerobic fermentation, indicating that bacterial phenotype regarding the hydrogen metabolism via the deletion of each operon is different with that of each single gene. Then, 4 triple hydrogenase operon mutants and one quadruple mutant were investigated to evaluate the hydrogen metabolism (hydrogen production and uptake) using glucose or glycerol as a substrate of hydrogen fermentation. With both the carbon sources, only Hyd-2 and Hyd-3 were able to produce hydrogen. Furthermore, all the hydrogenases showed hydrogen uptake activity. In addition, no hydrogen production and hydrogen uptake were detected in the quadruple mutant which does not have all 4 hydrogenases. Hydrogen production from Hyd-2 and Hyd-3 was further confirmed by complementing their operons in the cloning vector pBR322.     

Frontal cationic curing of epoxy resins in the presence of defoaming or expanding compounds

Thermal frontal polymerization was carried out with trimethylol propane triglycidyl ether using two different BF₃‐amine complexes, B‐950 and B‐110 from Leepoxy, as initiators for cationic polymerization. The amounts of filler (kaolin or fumed silica), defoaming, or expansion agents were varied to study how the compositions affected the front velocity, expansion, and flexural modulus of the resulting epoxy resins. The polymer produced with B‐950 initiator showed higher modulus than the polymers produced with B‐110. Moreover, fumed silica created stronger materials than kaolin. The presence of BYK as a defoamer or an expansion agent such as the Expancel #80 was also able to affect significantly the mechanical properties. differential scanning calorimetry studies indicated that the conversion was complete and that kaolin and silica increased the rate of reaction. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 40339.     

Microstructure and mechanical properties of ternary mortars with brick powder,glass powder,slag, fly ash,and limestone

The objective of this research is to study the effects produced by ternary binders which combine the addition of waste brick powder with fly ash, limestone, ground granulated blast furnace slag or waste glass powder in the microstructure and mechanical properties of mortars. In these ternary binders, the ordinary Portland cement was partially replaced by 10% of waste brick powder and 10% of another of the abovementioned additions. Mortars prepared with ordinary Portland cement without additions were also prepared. The microstructure was characterized with mercury intrusion porosimetry, electrical resistivity, and thermogravimetric analyses. Ultrasonic pulse velocity, compressive and flexural strengths were also determined. Mortars made using ternary binders with two active additions showed higher pore refinement and higher electrical resistivity at 250 days. Furthermore, their compressive strength and ultrasonic pulse velocity were relatively similar or even higher than that noted for reference specimens.     

Maskless Preparation of Spatially-Resolved Plasmonic Nanoparticles on Polydimethylsiloxane via In Situ Fluoride-Assisted Synthesis

Here, a fluoride-assisted route for the controlled in-situ synthesis of metal nanoparticles (NPs) (i.e., AgNPs, AuNPs) on polydimethylsiloxane (PDMS) is reported. The size and coverage of the NPs on the PDMS surface are modulated with time and over space during the synthetic process, leveraging the improved yield (10×) and faster kinetics (100×) of NP formation in the presence of F⁻ ions, compared to fluoride-free approaches. This enables the maskless preparation of both linear and step gradients and patterns of NPs in 1D and 2D on the PDMS surface. As an application in flexible plasmonics/photonics, continuous and step-wise spatial modulations of the plasmonic features of PDMS slabs with 1D and 2D AgNP gradients on the surface are demonstrated. An excellent spatially resolved tuning of key optical parameters, namely, optical density from zero to 5 and extinction ratio up to 100 dB, is achieved with AgNP gradients prepared in AgF solution for 12 minutes; the performance are comparable to those of commercial dielectric/interference filters. When used as a rejection filter in optical fluorescence microscopy, the AgNP-PDMS slabs are able to reject the excitation laser at 405 nm and retain the green fluorescence of microbeads (100 µm) used as test cases.     

CPSDebug: Automatic failure explanation in CPS models

International Journal on Software Tools for Technology Transfer - Debugging cyber-physical system (CPS) models is a cumbersome and costly activity. CPS models combine continuous and discrete...