Green tea polyphenol EGCg induces cell fusion via reactive oxygen species.

Background: Osteoclasts are multinucleated cells formed by macrophage cell fusion that are responsible for bone resorption. Previously, we found that treating osteoclastic progenitor cells with (-)-epigallocatechin gallate (EGCg) increased cell fusion. In this study, we aimed to identify factors involved in the cell fusion induced by EGCg. Methods: We hypothesized that EGCg-induced oxidative stress might be involved in cell fusion, and used macrophage cell line RAW264.7 cells. We evaluated cell fusion activity after adding the antioxidants N-acetyl-l-cysteine (NAC) or catalase in addition to EGCg. The mRNA expressions of genes related to cell fusion and bone resorption were quantified by real-time PCR. Finally, we added hydrogen peroxide and examined its effects on cell fusion and TRAP activity. Results: EGCg-induced cell fusion was strongly inhibited by the addition of NAC in a dose-dependent manner (EGCg with 5 mM NAC; decreased to 1.5%; p < 0.05), while the inhibitory effect of catalase was limited (EGCg with 500 U/mL catalase; decreased to 27.7%; p < 0.05). DC-STAMP expression was significantly upregulated by EGCg compared with the untreated group, and the upregulation was significantly suppressed by 5 mM NAC. Conversely, Nfatc1 and TRAP expression were not upregulated by EGCg. These results suggest that EGCg induces DC-STAMP expression via reactive oxygen species production, which regulates cell fusion but does not affect the osteoclastic pathway. Although treatment with hydrogen peroxide promoted the formation of multinucleated cells, no increase in TRAP activity was observed, which was similar to EGCg treatment. Conclusions: This study suggests that the increased cell fusion by EGCg may be induced by oxidative stress due to reactive oxygen species production.

Synthetic Nanocelluloses Fluorescently Responsible to Enzymatic Degradation.

Crystalline assemblies of cellulose and cellulose derivatives that can be synthetically produced by in vitro enzymatic reactions in a bottom-up manner have attracted increasing attention as chemically designable functional nanomaterials (e.g., synthetic nanocelluloses). In this study, we demonstrate the preparation and characterization of alkyl ß-celluloside assemblies loaded with fluorescent molecules, which are fluorescently responsible to the enzymatic degradation of the cellulose moieties. The fluorescent properties are afforded to the assemblies by their bilayer-structured nanosheet morphologies realized through the uptake of environmentally responsive fluorescent molecules (namely, Nile Red (NR)). Incubation of the NR-loaded n-octyl ß-celluloside (CEL-C8) assembly with cellulase resulted in decreases in the fluorescence intensities. This suggests that NR molecules were released into the aqueous phase through enzymatic degradation of the cellulose moieties of CEL-C8 molecules in the assembly. The fluorescence decrease rates were clearly dependent on the concentration and source of cellulase. Fluorescence decreases through enzymatic degradation were again observed in the presence of contaminant proteins. These observations revealed the high potential of alkyl ß-celluloside assemblies loaded with fluorescent molecules as fluorescently responsible cellulase substrates for cellulase detection assays by simply measuring changes in the fluorescence intensities. Moreover, the assemblies were revealed as carriers for the controlled release of loaded molecules triggered by enzymatic degradation.

Isolation and characterization of a Vibrio sp. strain MA3 associated with mass mortalities of the pearl oyster Pinctada fucata.

In the summers of 2019 and 2020, a previously undescribed disease occurred in both juvenile and adult shellfish, causing mass mortalities in cultured pearl production, characterized by the major symptom of extreme atrophy of the soft tissues, including the mantle. However, the causative organism was uncertain. We isolated Vibrio sp. strain MA3 from the mantles of diseased pearl oysters Pinctada fucata. Analyses of 16S rRNA gene and DNA gyrase sequence homologies and its biochemical and morphological characteristics suggested that strain MA3 is a new strain of Vibrio alginolyticus. In addition, a hemolysin gene (Vhe1) of strain MA3 was detected as one of the virulence factors, and the complete sequence was determined. BLAST searches showed that Vhe1 shares 99.8% nucleotide sequence identity with Vibrio alginolyticus strain A056 lecithin-dependent hemolysin (ldh) gene, complete cds. Experimental infection of healthy oysters via injection with strain MA3 indicated it could cause high mortalities of the typically affected oysters from which the strain was isolated. These results suggest that the newly isolated Vibrio sp. strain MA3 is a putative causal agent of the recent disease outbreaks in Akoya pearl oysters.

Control of parallel versus antiparallel molecular arrangements in crystalline assemblies of alkyl ß-cellulosides.

HYPOTHESIS: The precise control of parallel versus antiparallel molecular arrangements in synthetic assemblies of biorelated molecules is an attractive research focus from both scientific and technological viewpoints. However, little is known about cellulose-based synthetic assemblies. We hypothesized the existence of potential parameters, such as temperature, salt concentration, salt species, and solvent species, for controlling the molecular arrangement in assemblies of alkyl ß-cellulosides with different alkyl chain lengths. EXPERIMENTAL: The self-assembly of alkyl ß-cellulosides was triggered by neutralization-induced water insolubilization. The crystal structures of the cellulose moieties in the assemblies were characterized by attenuated total reflection-Fourier transform infrared absorption spectroscopy and wide-angle X-ray diffraction measurements. The morphologies of the assemblies were also characterized by scanning electron, atomic force, and transmission electron microscopy. FINDINGS: The temperature for the self-assembly, the concentration and species of inorganic salt in the self-assembly solution, and the solvent species (namely, the addition of water-miscible organic solvents into the self-assembly solution) strongly affected the molecular arrangement of the assemblies. The observations suggested that hydrophobic effects between the alkyl groups of the alkyl ß-cellulosides and/or interactions of the alkyl ß-cellulosides with solvent species were potential factors for controlling the molecular arrangement.

Structured liquids with interfacial robust assemblies of a nonionic crystalline surfactant.

HYPOTHESIS: The structuring of liquids, that is, the kinetic trapping of nonequilibrium shapes of liquid-liquid interfaces, shows great promise for various applications, especially all-liquid devices. The strategies reported thus far to stabilize such unstable states include interfacial jamming of large colloidal particles and interfacial coassembly of elaborate molecules and/or nanoparticles. However, the structuring of liquids using a simple molecular surfactant has not been sufficiently demonstrated. We hypothesized that a surfactant with strong intermolecular interactions would form interfacial assemblies that behave substantially as solid particles for the structuring of liquids. EXPERIMENTS: n-Octyl cello-oligosaccharide, a novel nonionic surfactant developed recently was explored as a candidate because of the ability of cello-oligosaccharides to form robust crystalline assemblies. Interfacial assembly of the nonionic crystalline surfactant was investigated for various water-organic solvent interfaces via pendant drop tensiometry and emulsification. FINDINGS: The crystalline surfactant was found to self-assemble and form a crystalline monolayer at water-organic solvent interfaces, allowing arrested shape changes of the liquid-liquid interfaces. Irregular-shaped liquid droplets were successfully created under various solution conditions, such as various organic solvents for the oil phase and the water phase with high ionic strengths and harsh pH values.

Mechanically robust crystalline monolayer assemblies of oligosaccharide-based amphiphiles on water surfaces.

Cellulose oligomers with a terminal alkyl group at the reducing end formed mechanically robust crystalline monolayers via self-assembly against water surfaces from aqueous solutions. The unique properties of the monolayers resulted in anisotropic deformation or manipulation of the aqueous solution droplets.