Smart polyion complex micelles for targeted intracellular delivery of PEGylated antisense oligonucleotides containing acid-labile linkages

A novel pH-sensitive and targetable antisense ODN delivery system based on multimolecular assembly into polyion complex (PIC) micelles of poly(L-lysine) (PLL) and a lactosylated poly(ethylene glycol)-antisense ODN conjugate (Lac-PEG-ODN) containing an acid-labile linkage (beta-propionate) between the PEG and ODN segments has been developed. The PIC micelles thus prepared had clustered lactose moieties on their peripheries and achieved a significant antisense effect against luciferase gene expression in HuH-7 cells (hepatoma cells), far more efficiently than that produced by the nonmicelle systems (ODN and Lac-PEG-ODN) alone, as well as by the lactose-free PIC micelle. In line with this pronounced antisense effect, the lactosylated PIC micelles showed better uptake than the lactose-free PIC micelles into HuH-7 cells; this suggested the involvement of an asialoglycoprotein (ASGP) receptor-mediated endocytosis process. Furthermore, a significant decrease in the antisense effect (27 % inhibition) was observed for a lactosylated PIC micelle without an acid-labile linkage (thiomaleimide linkage); this suggested the release of the active (free) antisense ODN molecules into the cellular interior in response to the pH decrease in the endosomal compartment is a key process in the antisense effect. Use of branched poly(ethylenimine) (B-PEI) instead of the PLL for PIC micellization led to a substantial decrease in the antisense effect, probably due to the buffer effect of the B-PEI in the endosome compartment, preventing the cleavage of the acid-labile linkage in the conjugate. The approach reported here is expected to be useful for the construction of smart intracellular delivery systems for antisense ODNs with therapeutic value.     

Synthesis of a photoimmobilizable histidine polymer for surface modification

A novel photoreactive polymer with histidine polar groups was synthesized through the copolymerization of two types of methacrylic acid, one carrying histidine groups and the other carrying azidoaniline groups. The polymer was photoimmobilized on polyester disks for surface modification. The effect of the surface modification on the hydrophilic and biofouling properties was investigated. Static contact angle measurements showed that the polymeric surface was modified to be comparatively hydrophilic in the polymer‐immobilized region. Micropattern immobilization was carried out with a photolithographic method. Atomic force microscopy measurements showed that the polymer was formed on the disks in response to ultraviolet irradiation. Protein adsorption was reduced on the polymer‐immobilized regions, and in those regions, spreading and adhesion of mammalian cells were reduced in comparison with that in nonimmobilized regions. In conclusion, a novel histidine‐containing polymer was photoreactively immobilized on a conventional polymer surface, and it had reduced interaction with proteins and cells. © 2008 Wiley Periodicals, Inc. J Appl Polym Sci, 2009     

Structural color films with lotus effects, superhydrophilicity, and tunable stop-bands

The structural blue color of a Morpho butterfly originates from the diffraction of light and interference effects due to the presence of the microstructures on the wing of the butterfly. Structural color on the surface of a damselfish reversibly changes between green and blue. Inspired by these creatures, we have been trying to prepare high-quality and functional structural color films. We describe our efforts in this Account. A useful technique to prepare such structural color films in colloidal solution is a "lifting" method, which allows us to quickly fabricate brilliant colloidal crystal films. The thicknesses of the films can be controlled by precisely adjusting the particle concentration and the lifting speed. Moreover, in order to prepare a complicated structure, we have used template methods. Indeed, we have successfully prepared the inverse structure of the wing of a Morpho butterfly with this technique. Initially, however, our structural color films had a whitish appearance due to the scattering of light by defects in the colloidal crystal film. Later, we were able to prepare a non-whitish structural color film by doping an appropriate dye in the colloidal particles to absorb the scattering light. In addition to the structural blue color, the wing of the Morpho butterfly has superhydrophobic properties. According to Wenzel's equation, the hydrophobic and hydrophilic properties are enhanced when the roughness of the hydrophobic and hydrophilic surface is increased, respectively. Based on this mechanism, we have successfully prepared structural color films with superhydrophobic properties, as well as with superhydrophilic properties. Another important property that can be seen in nature is tunable structural color, such as the color change that can be seen on the surface of a damselfish. In order to mimic such color change, we have developed several tunable structural color films. In particular, we have successfully prepared phototunable photonic crystals using photoresponsive azobenzene derivatives. In order to apply these structural color films, we developed a technique for patterning them by taking advantage of the wettability of the substrate surface. These materials can be used in the future for self-cleaning pigments and tunable photonic crystals.     

The alkylation of biphenyl over three-dimensional large pore zeolites: The influence of zeolite structure and alkylating agent on the selectivity for 4,4′-dialkylbiphenyl

Catalytic properties of three-dimensional zeolites, Y (FAU), Beta (BEA), and CIT-1 (CON) zeolites were examined in the alkylation, isopropylation, sec-butylation, and tert-butylation, of biphenyl (BP), and compared to those of H-mordenite (MOR). The selectivities for 4,4′-dialkylbiphenyl (4,4′-DABP) varied with the types of zeolite and of alkylating agent. FAU, BEA, and CON gave only low selectivities for 4,4′-diisopropylbiphenyl (4,4′-DIPB) in the isopropylation, and predominant isomers were bulky and thermodynamically unstable 2,x′-DIPB (2,2′-, 2,3′-, and 2,4′-) at lower temperatures, and bulky and thermodynamically stable 3,4′- and 3,3′-DIPB at higher temperatures: this is quite different from catalytic features over MOR, which gave 4,4′-DIPB with high selectivities at moderate temperatures. These results suggest that FAU, BEA, and CON have no shape-selective nature in the isopropylation, and that the reaction is principally controlled kinetically at lower temperatures, and thermodynamically at higher temperatures. The sec-butylation gave similar results to the isopropylation. Although the selectivities for 4,4′-di-sec-butylbiphenyl (4,4′-DSBB) were higher than those in the isopropylation, predominant isomers were 2,x′-DSBB (2,2′-, 2,3′-, and 2,4′-) at lower temperatures, and 3,4′- and 3,3′-DSBB at higher temperatures. The tert-butylation gave 4,4′-di-tert-butylbiphenyl (4,4′-DTBB) in moderate to high selectivities over all zeolites at moderate temperatures: the selectivity for 4,4′-DTBB was higher than 80% over BEA and CON; however, it still remained at 50% over FAU. FAU channels with super cages are too large for selective formation of 4,4′-DTBB.

From these results, it is concluded that the selectivity for 4,4′-DABP in the alkylation over MOR, FAU, BEA, and CON is determined by the exclusion of bulky isomers at their transition states, and that the exclusion is caused by the steric restriction at the transition states of bulky isomers by the zeolite channels.     

Development of gap sensing system for narrow gap laser welding

Narrow gap welding with an oscillation laser beam is one of the effective processes for thick plate welding. To put this welding process into practical manufacturing, a groove-sensing system using image processing for narrow gap welding with an oscillation laser beam is used. This developed system uses still images of the weld zone taken by a coaxial CMOS camera. It can recognize the position of the groove wall by analysing the brightness distribution in the still image. It can then control the oscillation width and the laser-irradiated area by calculating the groove width and the groove centre position. Some narrow gap welding experiments were performed to evaluate the performance of the developed system. The results revealed that the developed system is effective for narrow gap welding with an oscillation laser beam. Using this system, the narrow gap groove can be welded even if the groove width has changed during the welding process.     

Proton-conducting electrolyte membranes based on hyperbranched polymer with a sulfonic acid group for high-temperature fuel cells

The hyperbranched polymers (HBP-SA-Acs) with both a sulfonic acid group as a functional group and an acryloyl group as a cross-linker at terminals in different ratios of sulfonic acid group/acryloyl group (SO₃H/Ac) were successfully synthesized as a new thermally stable proton-conducting electrolyte. The cross-linked hyperbranched polymer electrolyte membranes (CL-HBP-SAs) were prepared by thermal polymerizations of the HBP-SA-Acs using benzoyl peroxide, and their ionic conductivities under dry condition and thermal properties were investigated. The ionic conductivities of the CL-HBP-SAs were found to be in the range of 2.2 × 10⁻⁴ to 3.3 × 10⁻⁶ S/cm, depending upon the SO₃H unit contents, at 150 °C under dry condition, and showed the Vogel-Tamman-Fulcher (VTF) type temperature dependence, indicating that proton transfer is cooperated by local polymer chain motion. All CL-HBP-SAs were thermally stable up to 260 °C, and they had suitable thermal stability as electrolyte membranes for the high-temperature fuel cells under dry condition. Fuel cell measurement using a single membrane electrode assembly cell with a cross-linked electrolyte membrane was successfully performed under non-humidified condition. It was demonstrated that applying the concept of dry polymer system to proton conduction is one possible approach toward high-temperature fuel cells.     

In situ measurement of lithium mass transfer during charging and discharging of a Ni–Sn alloy electrode

The charging (lithiation) and discharging (delithiation) of a Ni–Sn alloy electrode was examined by means of holographic interferometry and laser scanning confocal microscopy (LSCM). The developing concentration profile of Li⁺ ion during discharging (delithiation) of the Ni–Sn alloy electrode agreed reasonably well with the prediction based on transient diffusion theory, indicating that the cell configuration successfully suppressed natural convection due to the electrochemical reaction. The morphological variation of Ni–Sn alloy electrode during charge/discharge cycle could be observed very clearly by in situ LSCM. It was found that the contraction of active material during the discharge (delithiation) initiates cracks in the Ni–Sn alloy electrode.     

Optimization of antioxidant peptide production from grass carp sarcoplasmic protein using response surface methodology

The sarcoplasmic protein was extracted from the muscle of grass carp, a Chinese freshwater carp, and was hydrolyzed with five proteases to produce antioxidant peptides. Papain hydrolysate was found to have the greatest activity against lipid peroxidation. Response surface methodology (RSM) was applied to optimize the hydrolysis conditions (including enzyme to substrate ratio, time and temperature). The minimum EC₅₀ value (317.25 μg/mL) which signified the maximum antioxidant activity was obtained at an enzyme to substrate ratio of 0.79%, an incubation time of 5.69 h and an incubation temperature of 52.15 °C, which was in agreement with the predicted value (313.99 μg/mL) estimated by RSM within a 95% confidence interval. Also, it was found that moderate denaturation of the sarcoplasmic protein and a modest increase in the degree of hydrolysis promoted the antioxidant activities of the hydrolysates. Oligopeptides (< 3 kDa) contributed more to the antioxidant activity than polypeptides.     

Characteristics of transient vortices in the boundary layer on a rotating disk under orbital motion

The objective of this study is to experimentally examine the characteristics of transient vortices in the boundary layer on a disk undergoing both rotation and orbital motion. The velocity fluctuations on a rotating, orbiting disk （disk radius equal to orbital radius） are measured by the hot-wire method, and the effects of orbital motion on the transient vortices in the boundary layer are examined. When the ratio of the orbital speed to the speed of rotation is i-0.025, the interval of transient vortices depends on only the orbital radius, regardless of the directions of rota- tion and orbital motion. The rate of low-frequency disturbances increases as the orbital speed increases, and the vortices induced by these low-frequency disturbances travel over the disk and then develop in the region of in- creased velocity. Consequently, no vortices generated on a rotating disk under orbital motion are stationary rela- tive to the disk.