The scaffolding protein Flot2 regulates cytoneme-based transport of Wnt3 in gastric cancer

The Wnt/β-catenin signalling pathway regulates multiple cellular processes during development and many diseases, including cell proliferation, migration, and differentiation. Despite their hydrophobic nature, Wnt proteins exert their function over long distances to induce paracrine signalling. Recent studies have identified several factors involved in Wnt secretion, however, our understanding of how Wnt ligands are transported between cells to interact with their cognate receptors is still debated. Here, we demonstrate that gastric cancer cells utilise cytonemes to transport Wnt3 intercellularly to promote proliferation. Furthermore, we identify the membrane-bound scaffolding protein Flotillin-2 (Flot2), frequently overexpressed in gastric cancer, as a regulator of these cytonemes. Together with the Wnt co-receptor and cytoneme initiator Ror2, Flot2 determines the number and length of Wnt3 cytonemes in gastric cancer. Finally, we show that Flot2 is necessary for Wnt8a cytonemes during zebrafish embryogenesis, suggesting a conserved mechanism for Flot2-mediated Wnt transport on cytonemes in development and disease.

Physiochemically distinct SU-8 surfaces tailor Xylella fastidiosa cell-surface holdfast and colonization

SU-8 polymer is an excellent platform for diverse applications due to its high aspect ratio of micro/nanostructures fabrication and exceptional optical, chemical, and biocompatible properties. Although SU-8 has been often investigated for a variety of biological applications, how its surface properties influence both the interaction of bacterial cells with the substrate and its colonization is poorly understood. In this work, we tailor SU-8 nanoscale surface properties to investigate single cell motility, adhesion and successive colonization of a phytopathogenic bacteria, Xylella fastidiosa. Different surface properties of SU-8 thin films have been prepared using photolithography processing and oxygen plasma treatment. We found a significant difference in bacterial cell behavior and subsequent colonization on SU-8 as surface property changes. A larger density of carboxyl groups in hydrophilic plasma-treated SU-8 surfaces promotes faster cell motility in the earlier stage of the growth. The hydrophobic nature of pristine SU-8 surfaces has no trackable bacterial motility with 5 to 10 times more single cells adhered to surface than its plasma-treated counterpart. In fact, plasma-treated SU-8 samples suppressed bacterial adhesion, with surfaces showing less than 5% coverage. These results not only showcase that SU-8 surface properties can impact the bacterial behavior in a spatiotemporal manner, but also provide insights on the prominent ability of pathogens to evolve and adapt to different surface properties.

Preparation of Chitosan Particles as a Delivery System for Tetrahydrocurcumin: β-cyclodextrin Inclusive Compound for Colorectal Carcinoma

Background: Tetrahydrocurcumin is a hydrogenated active metabolite of curcumin that exhibits similar pharmacological effects to curcumin. However, its hydrophobic nature has limited its aqueous solubility and bioavailability. By incorporating the tetrahydro curcumin into β-cyclodextrin, its physiochemical property can be improved. Objective: To develop a chitosan composite loaded with tetrahydro curcumin inclusive complex, characterize the developed composites, and evaluate its effectiveness on cancer cells. Methods: Tetrahydrocurcumin was formulated into an inclusive complex with β-cyclodextrin in the ratio of 1:2 (Tetrahydrocurcumin: β-cyclodextrin). The tetrahydro curcumin inclusive complex loaded chitosan particles (THC IC-loaded CPs) were prepared using ionic gelation and later characterized using FTIR. Cytotoxicity of THC IC-loaded CPs in human colon cancer cells, Caco-2 cells, was examined using RTCA xCELLigence technology. The uptake of these particles by Caco-2 cells was also evaluated via fluorescing microscopy. Results: FTIR results confirmed the formation of the tetrahydrocurcumin inclusive complex and the loading of this complex into chitosan composites. The cytotoxic effect of THC IC-loaded CPs showed a dose-dependent relationship, and the IC50 found was 1.117mM and 0.959mM after 48 and 72 hours, respectively. THC IC-loaded CPs showed an immediate uptake by CaCo-2 cells, and the maximum uptake was observed after 1 hour of incubation. Conclusion: This study showed that THC IC-loaded CPs is a potential drug carrier to deliver tetrahydrocurcumin into cancer cells and able to produce a cytotoxic effect on cancer cells.

Engineered properties of polyurethane laden with beetroot and cerium oxide for cardiac patch application

The cardiac patch provides appropriate physicochemical properties and mechanical strength for the regeneration of damaged heart tissues. In this work, for the first-time, beetroot (BR) is blended with cerium oxide (CeO2) to produce nanofibrous polyurethane (PU) composite patch using electrospinning. The objective of this work is to fabricate the composite and examine its feasibility for cardiac patch applications. Morphological analysis revealed a dramatic reduction of fiber diameter of PU/BR (233 ± 175 nm) and PU/BR/CeO2 (331 ± 176 mm) compared to the pristine PU (994 ± 113 mm). Fourier transform infrared analysis (FTIR) analysis indicated functional peak intensities of the newly formed composite PU/BR and PU/BR/CeO2 were not similar to PU. The addition of beetroot rendered PU/BR hydrophilic (86° ± 2), whereas PU/BR/CeO2exhibited hydrophobic nature (99° ± 3). Atomic force microscopy (AFM) analysis depicted the reduced surface roughness of the PU/BR (312 ± 12 nm) and PU/BR/CeO2 (390 ± 125 nm) than the pristine PU (854 ± 32 nm). The incorporation of beetroot and CeO2 into PU enhanced the tensile strength compared with the pristine PU. The blood clotting time of PU/BR (APTT-204 ± 3 s and PT-103 ± 2 s) and PU/BR/CeO2 (APTT-205 ± 3 s and PT-105 ± 2s) were delayed significantly than the pristine PU(APTT-176 ± 2 s and PT-94 ± 2 s) as revealed in the coagulation study. Further, hemolysis assay showed the less toxic nature of the fabricated composites than the pristine PU. Hence, it can be inferred that the advanced physicochemical and blood compatible properties of electrospun PU/BR and PU/BR/CeO2 nanocomposite can be engineered successfully for cardiac patch applications.

Study of Dissolution Kinetics of Mefenamic Acid Solid Dispersion with Polyvinylpyrrolidone

The low solubility of a biologically active substance in an aqueous medium is often the main reason for the reduced therapeutic effect of drugs. The most common approach to solve this problem is to obtain a watersoluble salt of the active substance and an appropriate preparatory formulation based on it. In this case, the solubility of the obtained compound in hydrophobic systems decreases dramatically, which decreases the rate of transmembrane transport and changes the pharmacokinetic laws of the process. In practice, not only the dependence of the therapeutic effect on the salt compound properties, but also a complete loss of the drug active ingredient activity can be observed. The use of biologically active compound solid dispersions in watersoluble polymers is the most promising approach to increase the therapeutic effect of drugs while maintaining the hydrophobic nature of the active component, to reduce the dose load on the patient’s body and obtain prolonged action. In experiments we obtained solid dispersions of mefenamic acid in polyvinylpyrrolidone and studied kinetic regularities of solubility of this promising drug form in aqueous solution of phosphate buffer. By means of mathematical modelling it was found that the phenomenon under study is well described by Ritger --- Peppas model, which considers diffusion of biologically active component into solution according to Fick's law with possible influence on mass transfer at swelling and degradation of polymer matrix

Adsorptive Capture of Ionic and Non-Ionic Pollutants Using a Versatile Hybrid Amphiphilic-Nanomica

A versatile, functional nanomaterial for the removal of ionic and non-ionic pollutants is presented in this work. For that purpose, the high charge mica Na-4-Mica was exchanged with the cationic surfactant (C16H33NH(CH3)2)+. The intercalation of the tertiary amine in the swellable nano-clay provides the optimal hydrophilic/hydrophobic nature in the bidimensional galleries of the nanomaterial responsible for the dual functionality. The organo-mica, made by functionalization with C16H33NH3+, was also synthesized for comparison purposes. Both samples were characterized by X-ray diffraction techniques and transmission electron microscopy. Then, the samples were exposed to a saturated atmosphere of cyclohexylamine for two days, and the adsorption capacity was evaluated by thermogravimetric measurements. Eu3+ cations served as a proof of concept for the adsorption of ionic pollutants in an aqueous solution. Optical measurements were used to identify the adsorption mechanism of Eu3+ cations, since Eu3+ emissions, including the relative intensity of different f–f transitions and the luminescence lifetime, can be used as an ideal spectroscopic probe to characterize the local environment. Finally, the stability of the amphiphilic hybrid nanomaterial after the adsorption was also tested.

Molecular dynamics simulations reveal the selectivity mechanism of structurally similar agonists to TLR7 and TLR8

TLR7 and TLR8 are key members of the Toll-like receptor family, playing crucial roles in the signaling pathways of innate immunity, and thus become attractive therapeutic targets of many diseases including infections and cancer. Although TLR7 and TLR8 show a highly degree of sequence homology, their biological response to small molecule binding is very different. Aiming to understand the mechanism of selective profiles of small molecule modulators against TLR7 and TLR8, we carried out molecular dynamic simulations on three imidazoquinoline derivatives bound to the receptors separately. They are Resiquimod (R), Hybrid-2 (H), and Gardiquimod (G), selective agonists of TLR7 and TLR8. Our MD trajectories indicated that in the complex of TLR7-R and TLR7-G, the two chains forming the TLR7 dimer tended to remain “open” conformation, while the rest systems maintained in the closed format. The agonists R, H, and G developed conformational deviation mainly on the aliphatic tail. Furthermore, we attempted to quantify the selectivity between TLR7 and TLR8 by binding free energies via MM-GBSA method. It showed that the three selected modulators were more favorable for TLR7 than TLR8, and the ranking from the strongest to the weakest was H, R and G, aligning well with experiment data. In the TLR7, the flexible and hydrophobic aliphatic side chain of H has stronger van der Waals interactions with Val381 and Phe351 but only pick up interaction with one amino acid residue i.e. Tyr353 of TLR8. Unsurprisingly, the positively charged side chain of G has less favor interaction with Ile585 of TLR7 and Val573 of TLR8 explaining G is weak agonist in both TLR7 and TLR8. All three imidazoquinolines can form stable hydrogen bonds with Asp555 of TLR7 and the corresponding Asp543 of TLR8. In brief, the set of total 400ns MD studies sheds light on the potential selective mechanisms of agonists towards TLR7 and TLR8, indicating the van der Waals interaction as the driving force for the agonists binding, thus provides us insights for more potent and selective modulators to cooperate with the hydrophobic nature of the binding pocket.

A Note of Caution: Gramicidin Affects Signaling Pathways Independently of Its Effects on Plasma Membrane Conductance

Gramicidin is a thoroughly studied cation ionophore widely used to experimentally manipulate the plasma membrane potential (PMP). In addition, it has been established that the drug, due to its hydrophobic nature, is capable of affecting the organization of membrane lipids. We have previously shown that modifications in the plasma membrane potential of epithelial cells in culture determine reorganizations of the cytoskeleton. To elucidate the molecular mechanisms involved, we explored the effects of PMP depolarization on some putative signaling intermediates. In the course of these studies, we came across some results that could not be interpreted in terms of the properties of gramicidin as an ionic channel. The purpose of the present work is to communicate these results and, in general, to draw attention to the fact that gramicidin effects can be misleadingly attributed to its ionic or electrical properties. In addition, this work also contributes with some novel findings of the modifications provoked on the signaling intermediates by PMP depolarization and hyperpolarization.

Modulation of Conductivity of Alginate Hydrogels Containing Reduced Graphene Oxide through the Addition of Proteins

Modifying hydrogels in order to enhance their conductivity is an exciting field with applications in cardio and neuro-regenerative medicine. Therefore, we have designed hybrid alginate hydrogels containing uncoated and protein-coated reduced graphene oxide (rGO). We specifically studied the adsorption of three different proteins, BSA, elastin, and collagen, and the outcomes when these protein-coated rGO nanocomposites are embedded within the hydrogels. Our results demonstrate that BSA, elastin, and collagen are adsorbed onto the rGO surface, through a non-spontaneous phenomenon that fits Langmuir and pseudo-second-order adsorption models. Protein-coated rGOs are able to preclude further adsorption of erythropoietin, but not insulin. Collagen showed better adsorption capacity than BSA and elastin due to its hydrophobic nature, although requiring more energy. Moreover, collagen-coated rGO hybrid alginate hydrogels showed an enhancement in conductivity, showing that it could be a promising conductive scaffold for regenerative medicine.