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.     

Understanding the Self-Assembly of Cellulose Nanocrystals—Toward Chiral Photonic Materials

Over millions of years, animals and plants have evolved complex molecules and structures that endow them with vibrant colors. Among the sources of natural coloration, structural color is prominent in insects, bird feathers, snake skin, plants, and other organisms, where the color arises from the interaction of light with nanoscale features rather than absorption from a pigment. Cellulose nanocrystals (CNCs) are a biorenewable resource that spontaneously organize into chiral nematic liquid crystals having a hierarchical structure that resembles the Bouligand structure of arthropod shells. The periodic, chiral nematic organization of CNC films leads them to diffract light, making them appear iridescent. Over the past two decades, there have been many advances to develop the photonic properties of CNCs for applications ranging from cosmetics to sensors. Here, the origin of color in CNCs, the control of photonic properties of CNC films, the development of new composite materials of CNCs that can yield flexible photonic structures, and the future challenges in this field are discussed. In particular, recent efforts to make flexible photonic materials using CNCs are highlighted.     

Translational Read-Through Therapy of RPGR Nonsense Mutations

X-chromosomal retinitis pigmentosa (RP) frequently is caused by mutations in the retinitis pigmentosa GTPase regulator (RPGR) gene. We evaluated the potential of PTC124 (Ataluren, Translama^TM) treatment to promote ribosomal read-through of premature termination codons (PTC) in RPGR. Expression constructs in HEK293T cells showed that the efficacy of read-through reagents is higher for UGA than UAA PTCs. We identified the novel hemizygous nonsense mutation c.1154T > A, p.Leu385* ({"type":"entrez-nucleotide","attrs":{"text":"NM_000328.3","term_id":"1677500221"}}NM_000328.3) causing a UAA PTC in RPGR and generated patient-derived fibroblasts. Immunocytochemistry of serum-starved control fibroblasts showed the RPGR protein in a dot-like expression pattern along the primary cilium. In contrast, RPGR was no longer detectable at the primary cilium in patient-derived cells. Applying PTC124 restored RPGR at the cilium in approximately 8% of patient-derived cells. RT-PCR and Western blot assays verified the pathogenic mechanisms underlying the nonsense variant. Immunofluorescence stainings confirmed the successful PTC124 treatment. Our results showed for the first time that PTC124 induces read-through of PTCs in RPGR and restores the localization of the RPGR protein at the primary cilium in patient-derived cells. These results may provide a promising new treatment option for patients suffering from nonsense mutations in RPGR or other genetic diseases.     

Physical filtration efficiency analysis of a polyaniline hybrid composite filter with graphite oxide for particulate matter 2.5

Particulate matter (PM) 2.5 pollution is a prevalent environmental and public health issue that has raised serious global concerns. Because standard heating, ventilation, and air conditioning filters are incapable of filtering out PM 2.5 particles efficiently, different methods of PM 2.5 filtration, such as physical filtration and electrostatic filtration, are under investigation to develop a filter with a high filtration efficiency and a low pressure drop. According to various studies, pressure drop has a significant influence on the filtration efficiency. An equation for the theoretical trend was generated based on the composite data gathered from similar filtration studies and was used to evaluate the relationship between pressure drop and filtration efficiency. Here, the theoretical equation indicated that the filtration efficiency increased as the pressure drop on a filter increased until 0.01 psi where the efficiency remained near constant at approximately 99.9%. In this study, we introduce a graphite oxide (GO) and polyaniline (PANi) composite hybrid filter in order to create a low-pressure (1.2 psi) drop filter. By adding GO flakes to the PANi matrix, we not only produced a highly permeable filter while allowing continuous gas flow, but also achieved a remarkable and highly effective PM 2.5 filter with a filtration efficiency of 99.7 ± 0.08%.     

Utilization of different tall oils for improving the water resistance of cellulosic fibers

This study was conducted to assess the effect of the pulping by-products crude tall oil (CTO), distilled tall oil (DTO), and tall oil fatty acid (TOFA) on dynamic water vapor sorption behavior, interfiber strength, and thermal stability of cellulosic paper-sheets. The results were compared against those obtained in cellulose papers treated with the conventional petroleum-derived hydrophobic agent hydrowax and in untreated ones. The tall oil treatments caused strong reduction in equilibrium moisture content of the paper-sheets during adsorption and desorption runs. The same trend was noticed for the hydrowax-treated papers, however, it was less pronounced than the CTO-treated and DTO-treated samples in the relative humidity range of 75–95%. The sorption hysteresis was considerably decreased after the treatments. The ultimate dry-tensile strengths of the paper-sheets were significantly reduced by TOFA and hydrowax treatments, while CTO and DTO showed comparable strength as that of untreated control. The ultimate wet-strengths of the paper-sheets were improved after the treatments. The thermal stability of the specimens was improved by the tall oil treatments, and the hydrowax-treated samples illustrated lower degradation temperature than the untreated control. The results are promising for the use of tall oils as alternative hydrophobic agents of cellulosic fiber-based products, such as wood panels and paper packaging. © 2018 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2019 , 136, 47303.     

Impact of Intrinsic Muscle Weakness on Muscle–Bone Crosstalk in Osteogenesis Imperfecta

Bone and muscle are highly synergistic tissues that communicate extensively via mechanotransduction and biochemical signaling. Osteogenesis imperfecta (OI) is a heritable connective tissue disorder of severe bone fragility and recently recognized skeletal muscle weakness. The presence of impaired bone and muscle in OI leads to a continuous cycle of altered muscle–bone crosstalk with weak muscles further compromising bone and vice versa. Currently, there is no cure for OI and understanding the pathogenesis of the skeletal muscle weakness in relation to the bone pathogenesis of OI in light of the critical role of muscle–bone crosstalk is essential to developing and identifying novel therapeutic targets and strategies for OI. This review will highlight how impaired skeletal muscle function contributes to the pathophysiology of OI and how this phenomenon further perpetuates bone fragility.     

Antioxidant Activity of Seaweed Extracts: In Vitro Assays,Evaluation in 5 % Fish Oil‐in‐Water Emulsions and Characterization

In this study the antioxidant activity of absolute ethanol, 50 % ethanol and water extracts of two species of seaweeds, namely Fucus serratus and Polysiphonia fucoides, were evaluated both in in vitro assays and in 5 % fish oil‐in‐water (o/w) emulsions. The 50 % ethanolic extracts of P. fucoides showed higher antioxidant activity both in in vitro assays and in 5 % oil‐in‐water emulsion in the presence or absence of iron. In spite of the higher phenolic content and very good antioxidant activity in some of the in vitro assays, the absolute ethanol extracts of both the species showed a pro‐oxidative tendency in 5 % fish oil‐in‐water emulsion in the presence or absence of iron. In order to investigate the reason for the higher antioxidant activity of 50 % ethanolic extracts of P. fucoides, these extracts were further fractionated into polyphenol‐rich, protein‐rich, polysaccharide‐rich and low‐molecular‐weight fractions. These fractions were tested both in in vitro and in 5 % oil‐in‐water emulsions. The results of the present study showed that the main effect was due to the phenolic compounds. In conclusion, the 50 % ethanolic extracts of P. fucoides can be a potential source of natural antioxidants as these extracts have antioxidant activities similar to those of synthetic antioxidants such as BHT.     

Thioether Analogues of Disulfide‐Bridged Cyclic Peptides Targeting Death Receptor 5: Conformational Analysis,Dimerisation and Consequences for Receptor Activation

Cyclic peptides containing redox‐stable thioether bridges might provide a useful alternative to disulfide‐bridged bioactive peptides. We report the effect of replacing the disulfide bridge with a lanthionine linkage in a 16‐mer cyclic peptide that binds to death receptor 5 (DR5, TRAIL‐R2). Upon covalent oligomerisation, the disulfide‐bridged peptide has previously shown similar behaviour to that of TNF‐related apoptosis inducing ligand (TRAIL), by selectively triggering the DR5 cell death pathway. The structural and biological properties of the DR5‐binding peptide and its desulfurised analogue were compared. Surface plasmon resonance (SPR) data suggest that these peptides bind DR5 with comparable affinities. The same holds true for dimeric versions of these peptides: the thioether is able to induce DR5‐mediated apoptosis of BJAB lymphoma and tumorigenic BJELR cells, albeit to a slightly lower extent compared to its disulfide homologue. NMR analysis revealed subtle variation in the conformations of the two peptides and suggests that the thioether peptide is slightly less folded than its disulfide homologue. These observations could account for the different capability of the two dimers to cluster DR5 receptors on the cell surface and to trigger apoptosis. Nevertheless, our results suggest that the thioether peptide is a potential candidate for evaluation in animal models.