Magnetically and Near‐Infrared Light‐Powered Supramolecular Nanotransporters for the Remote Control of Enzymatic Reactions

Cancer is one of the primary causes of death worldwide. A high‐precision analysis of biomolecular behaviors in cancer cells at the single‐cell level and more effective cancer therapies are urgently required. Here, we describe the development of a magnetically‐ and near infrared light‐triggered optical control method, based on nanorobotics, for the analyses of cellular functions. A new type of nanotransporters, composed of magnetic iron nanoparticles, carbon nanohorns, and liposomes, was synthesized for the spatiotemporal control of cellular functions in cells and mice. Our technology will help to create a new state‐of‐the‐art tool for the comprehensive analysis of “real” biological molecular information at the single‐cell level, and it may also help in the development of innovative cancer therapies.     

Unidirectional crystallization of charged colloidal silica due to the diffusion of a base

Dilute aqueous dispersions of charged colloidal silica (particle volume fraction = approximately 0.03-0.04, particle diameter = 110 nm) exhibit unidirectional crystal growth due to the diffusion of a weak base, pyridine (Py). Similar diffusion-crystallization is enabled by a salt of a weak acid and a strong base, sodium hydrogen carbonate (NaHCO3). The resulting crystals consist of columnar (or cubic) crystal grains with a maximum height of a few centimeters and a maximum width of 1 cm. The crystal growth process is attributed to a combination of (i) the diffusion of Py or NaHCO3 accompanied by a charging reaction of the silica particles and (ii) the charge-induced crystallization of the silica colloids. Theoretical growth curves based on the reaction-diffusion model for the case of Py were in good agreement with the observed curves. We also report the immobilization of the resulting large crystals by using a polymer hydrogel matrix.     

Adsorption kinetics of carcinogens to DNA liquid crystalline gel beads

Adsorption behaviors of acridine orange (AO) and biphenyl (BP) to DNA liquid crystalline gel (LCG) beads in aqueous dispersing solution have been studied theoretically and experimentally. A theoretical consideration based on nonequilibrium thermodynamics predicted that the time course of the adsorption process is expressed with a scaled equation, and a scaled number of adsorbed carcinogen molecules ? is expressed with the square root of a scaled immersion time t, ? proportional, variant square root t at early stage, whereas it is expressed with a power law function 1 - ? proportional, variant (te - t)3/2 for ?0 > 1 and an exponential equation ?0 - ? proportional, variant e-t/alpha tau0 for ?0 > 1 at later stages of adsorption. Here, ?0 is the ratio of the initial number of carcinogen molecules in the dispersing solution to the number of the sites of adsorption of carcinogen molecules in the beads, te is the scaled equilibrium time of adsorption, tau0 is a time constant for adsorption, and alpha is a constant. Observed adsorption processes for AO were well expressed by the predicted ones, and the fitting parameters ?0 and tau0 increased with increasing cobalt chloride concentration CCo used for preparation of the beads, and both saturated above CCo > or = 400 mM for the adsorption of AO, whereas the adsorption processes for BP were expressed with the square root function. These results indicate that (1) the adsorption process at early stage is explained by diffusion-limited binding of the carcinogen molecules to DNA beads, and the time range of the early stage depends on the solubility (the solubility of AO in water is high whereas that of BP is low); and (2) the process at later stages depends on the balance of the numbers of adsorption sites and carcinogen molecules.     

In Vitro Analyses of Spinach-Derived Opioid Peptides,Rubiscolins: Receptor Selectivity and Intracellular Activities through G Protein- and β-Arrestin-Mediated Pathways

Activated opioid receptors transmit internal signals through two major pathways: the G-protein-mediated pathway, which exerts analgesia, and the β-arrestin-mediated pathway, which leads to unfavorable side effects. Hence, G-protein-biased opioid agonists are preferable as opioid analgesics. Rubiscolins, the spinach-derived naturally occurring opioid peptides, are selective δ opioid receptor agonists, and their p.o. administration exhibits antinociceptive effects. Although the potency and effect of rubiscolins as G-protein-biased molecules are partially confirmed, their in vitro profiles remain unclear. We, therefore, evaluated the properties of rubiscolins, in detail, through several analyses, including the CellKey^TM assay, cADDis^® cAMP assay, and PathHunter^® β-arrestin recruitment assay, using cells stably expressing µ, δ, κ, or µ/δ heteromer opioid receptors. In the CellKey^TM assay, rubiscolins showed selective agonistic effects for δ opioid receptor and little agonistic or antagonistic effects for µ and κ opioid receptors. Furthermore, rubiscolins were found to be G-protein-biased δ opioid receptor agonists based on the results obtained in cADDis^® cAMP and PathHunter^® β-arrestin recruitment assays. Finally, we found, for the first time, that they are also partially agonistic for the µ/δ dimers. In conclusion, rubiscolins could serve as attractive seeds, as δ opioid receptor-specific agonists, for the development of novel opioid analgesics with reduced side effects.     

Formation of fractal porous silica by hydrolysis of TEOS in a bicontinuous microemulsion

Mesoporous silica particles have been prepared by hydrolysis of TEOS (Si(OC₂H₅)₄) in bicontinuous microemulsions containing polyoxyethylene (POE) dodecylether, isooctane and water. TEOS was dissolved in a continuous water phase and hydrolyzed by the dispersed water at around the phase inversion temperature (60°C). Undulating solid materials with layered mesostructures were produced from middle-phase microemulsions in the three phase region (o/w=0.2–0.7). On the other hand, the solids obtained from the lower aqueous phase in the three phase region were found to have a heterogeneous disordered structure. Measurements of the fractal dimensions were performed in the macropore region using a box-counting method for the outline of the SEM texture. We found that the macropore size distribution in the particles prepared from the middle-phase microemulsion follows the fractal rule with a dimension of 1.7. From the results of nitrogen adsorption/desorption curves on the silica, a steep increase in the adsorption amounts was observed at a relative pressure below 0.2, and adsorption/desorption hysteresis was also observed at a relative pressure between 0.3 and 0.5. These studies suggest that the silica synthesized in the bicontinuous microemulsion mesostructure has a very broad size range from micro to macropores with a fractal distribution.     

Thermoreversible gelation and phase separation in aqueous methyl cellulose solutions

We derived typical phase diagrams for aqueous solutions of methyl cellulose (MC) of different molecular weights via micro‐differential scanning calorimetry, small‐angle X‐ray scattering, and visual inspection. The phase diagrams showed the cooccurrence of gelation and phase separation and qualitatively agreed with the theoretically calculated diagrams. The sol–gel transition line and phase separation line of a lower critical solution point type shifted toward lower temperatures and lower concentrations with an increase in the MC molecular weight. The sol–gel transition line intersected at a temperature higher than the critical point of the phase separation; therefore, both sol–gel phase separation and gel–gel phase separation were possible, depending on the temperature. Specifically, through visual inspection of a high molecular weight MC sample in the critical temperature region, we observed phase separation into two coexisting gels with different polymer concentrations. © 2000 John Wiley & Sons, Inc. J Polym Sci B: Polym Phys 39: 91–100, 2001     

Electrostatic Assembly of Dendrimer Electrolytes: Negatively and Positively Charged Dendrimer Porphyrins

Carboxylate and ammonium functionalities—32 of each— facilitate electrostatic interaction between oppositely charged dendrimer electrolytes and porphyrin cores, which leads in protic media to a supramolecular, fluorescence-active aggregate of two chromophores that communicate and have a predictable topology (see drawing on the right; A=acceptor, D=donor). In contrast to the complexation of linear polyelectrolytes, the contact area for the two dendrimer molecules is very limited, just as one would expect for an assembly of spherical molecules.