The novel functional nucleic acid iRed effectively regulates target genes following cytoplasmic delivery by faint electric treatment

An intelligent shRNA expression device (iRed) contains the minimum essential components needed for shRNA production in cells, and could be a novel tool to regulate target genes. However, general delivery carriers consisting of cationic polymers/lipids could impede function of a newly generated shRNA via electrostatic interaction in the cytoplasm. Recently, we found that faint electric treatment (fET) of cells enhanced delivery of siRNA and functional nucleic acids into the cytoplasm in the absence of delivery carriers. Here, we examined fET of cells stably expressing luciferase in the presence of iRed encoding anti-luciferase shRNA. Transfection of lipofectamine 2000 (LFN)/iRed lipoplexes showed an RNAi effect, but fET-mediated iRed transfection did not, likely because of the endosomal localization of iRed after delivery. However, fET in the presence of lysosomotropic agent chloroquine significantly improved the RNAi effect of iRed/fET to levels that were higher than those for the LFN/iRed lipoplexes. Furthermore, the amount of lipid droplets in adipocytes significantly decreased following fET with iRed against resistin in the presence of chloroquine. Thus, iRed could be a useful tool to regulate target genes following fET-mediated cytoplasmic delivery with endosomal escape devices.     

Syntheses of sodium 2-(N-alkyl-N-methylamino)ethanephosphates and their physicochemical properties

A series of new amphoteric surfactants, sodium salts of 2-(N-alkyl-N-methylamino)ethanephosphates (alkyl: n-dodecyl, n-tetradecyl and n-hexadecyl) were prepared by reacting alkylbromides with N-methylaminoethanol, followed by addition of phosphoric acid groups and neutralization with sodium hydroxide. The structures of these compounds were confirmed by spectroscopy and elemental analysis. Physical properties such as Krafft point, isoelectric point, critical micelle concentration, occupation area of molecule at surface, foaming power and the effect of pH on surface activity were evaluated. These surfactants were found to exhibit zwitterionic characteristics in the pH range from approximately 5 to 9 and demonstrated good surface-active properties over a wide pH range (pH 4~10). Large occupation areas of the molecule at a surface in comparison with sodium dodecyl sulfate was noted as a characteristic feature of these surfactants.     

Metabolities of dietary triacylglycerol and diacylglycerol during the digestion process in rats

The present study investigated the metabolic fate of dietary TAG and DAG and also their digestion products in the stomach and small intestine. A diet containing 10% TAG or DAG oil, enriched in 1,3-DAG, was fed to Wistar rats ad libitum for 9 d. After 18 h of fasting, each diet was re-fed ad libitum for 1 h. The weights of the contents of the stomach and small intestine were measured, and the acylglycerol and FFA levels were analyzed by GC at 0, 1, and 4 h after the 1-h re-feeding. The amounts of re-fed diet ingested and the gastric and small intestinal content were not different between the two diet groups. In the TAG diet group, the main products were TAG and DAG, especially 1(3),2-DAG. In addition, 1,3-DAG and 1(3)-MAG were present in the stomach, and the 1,3-DAG levels increased over time after the re-feeding period. In the DAG diet group, the main products in the stomach were DAG, MAG, FFA, and TAG. There were significantly greater amounts of 1,3-DAG, 1(3)-MAG, and FFA in the DAG diet group in the stomach compared with the TAG diet group. The amount of FFA in the stomach relative to the amount of ingested TAG plus DAG in the DAG diet group was higher than that in the TAG diet group. Acylglycerol and FFA levels were considerably lower in the small intestine than in the stomach. These results indicate that, in the stomach, where acyl migration might occur, the digestion products were already different between TAG and DAG oil ingestion, and that DAG might be more readily digested by lingual lipase compared with TAG. Furthermore, almost all of the dietary lipid was absorbed, irrespective of the structure of the acylglycerol present in the small intestine.     

Development of Low Temperature SQUID Gradiometer Array for Metallic Magnetic Microcalorimeters

We report on the performance of two types of SQUID gradiometers developed for the readout of magnetic calorimeters. Our previously developed low dissipation SQUID gradiometer optimized for low temperature operation has demonstrated the flux noise level of under a magnetic field of 2.5 mT and 150 mK. With a cylindrical Au:Er paramagnetic sensor mounted inside the octagonal pickup washer of the SQUID gradiometer, we succeeded in detecting X-ray signals. However, our achieved energy resolution was 47.2±2.1 eV at 5.9 keV limited by the high operating temperature of 150 mK and by a magnetic field, small for that temperature, due to the limited critical current of the field coils. Based on these results, we fabricated new arrays of SQUID gradiometer by tuning the line width and the number of turns of the field coils and shunt resistance to realize a lower noise level and a larger magnetic field. Furthermore, arrays of SQUID gradiometer with meander patterned pickup washer was fabricated which provides a stronger coupling between the paramagnetic sensor and the pickup washer, and a larger magnetic field at the sensor.      

Characterization and properties of iron-incorporated gismondine prepared at 80°C

Iron-incorporated zeolites were successfully synthesized at a low temperature such as 80°C by choosing appropriate starting materials and characterized by inductively coupled plasma atomic emission spectrometry (ICP-AES), wide-angle X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FT-IR), and magnetic susceptibility. ICP-AES showed␣that Fe component can be readily incorporated␣up to a maximum extent of Fe substitution, Fe/(Fe + Al) × 100 = 22.7%. XRD measurements suggested that the zeolites obtained have a crystal structure of gismondine type. The characterizations identified that the Fe component present in the products is all incorporated into the zeolite framework. The ammonia and water desorption profiles were compared for Fe-free and 22.7% Fe-zeolites ion-exchanged for NH₄⁺ by means of TG-MS and DSC. The ammonia desorption peak temperatures considerably shifted toward lower temperatures by the introduction of Fe, suggesting decreased solid acidity. DSC thermograms of the as-synthesized gismondines revealed that they do not contain free water (i.e., water not coordinated to cations) in the pores irrespective of the Fe content. The enhanced catalytic reactivity of the Fe-incorporated gismondines was also confirmed from the decomposition of hydrogen peroxide. An apparent activation energy of 43 kJ mol⁻¹ was obtained independent of the Fe contents in zeolites. This value was much lower than 70 kJ mol⁻¹ for the same reaction in the homogeneous solution containing iron alum as a reference sample.     

Syntheses of 2-(N-alkyl-N,N-dimethylammonio)ethyl hydrogen phosphates and their physicochemical properties

A series of new amphoteric surfactants, having a quaternary ammonium group and a phosphoric acid group, 2-(N-alkyl-N,N-dimethylammonio)ethyl hydrogen phosphates (alkyl: dodecyl, tetradecyl, hexadecyl), were prepared by introducing a phosphoric acid group into N-alkyl-N,N-dimethyl-(N-2-hydroxyethyl) ammoniumiodide, followed by neutralization with sodium hydroxide and removal of inorganic salts. By the evaluation of the physicochemical properties, it was found that the phosphobetaines behave like “nonionic” surfactant in the zwitterionic region, having very small CMC values in comparison with sodium 2-(N-alkyl-N-methylamino)ethyl hydrogen phosphates. Additionally, similar trends were also observed in the experiments from the point of the effect of electrolyte and temperature on CMC, respectively. It should be considered that the hydrophobicity of the surfactant molecule is increased by the electronic interaction between the quaternary ammonium group and a phosphoric acid group. On the other hand, the phosphobetaines gave the smaller values of the occupied area per molecule at the air/water interface than sodium 2-(N-alkyl-N-methylamino)ethyl hydrogen phosphates. Therefore, it is clear that the phosphobetaines have higher surface-active properties than the amphoteric surfactants.     

The E3 Ubiquitin-Protein Ligase RNF4 Promotes TNF-α-Induced Cell Death Triggered by RIPK1

Receptor-interacting protein kinase 1 (RIPK1) is a key component of the tumor necrosis factor (TNF) receptor signaling complex that regulates both pro- and anti-apoptotic signaling. The reciprocal functions of RIPK1 in TNF signaling are determined by the state of the posttranslational modifications (PTMs) of RIPK1. However, the underlying mechanisms associated with the PTMs of RIPK1 are unclear. In this study, we found that RING finger protein 4 (RNF4), a RING finger E3 ubiquitin ligase, is required for the RIPK1 autophosphorylation and subsequent cell death. It has been reported that RNF4 negatively regulates TNF-α-induced activation of the nuclear factor-κB (NF-κB) through downregulation of transforming growth factor β-activated kinase 1 (TAK1) activity, indicating the possibility that RNF4-mediated TAK1 suppression results in enhanced sensitivity to cell death. However, interestingly, RNF4 was needed to induce RIPK1-mediated cell death even in the absence of TAK1, suggesting that RNF4 can promote RIPK1-mediated cell death without suppressing the TAK1 activity. Thus, these observations reveal the existence of a novel mechanism whereby RNF4 promotes the autophosphorylation of RIPK1, which provides a novel insight into the molecular basis for the PTMs of RIPK1.     

The effect of abrading and cutting instruments on machinability of dental ceramics

The aim was to investigate the effect of machining instruments on machinability of dental ceramics. Four dental ceramics, including two zirconia ceramics were machined by three types (SiC, diamond vitrified, and diamond sintered) of wheels with a hand-piece engine and two types (diamond and carbide) of burs with a high-speed air turbine. The machining conditions used were abrading speeds of 10,000 and 15,000?r.p.m. with abrading force of 100?gf for the hand-piece engine, and a pressure of 200?kPa and a cutting force of 80?gf for the air-turbine hand-piece. The machinability efficiency was evaluated by volume losses after machining the ceramics. A high-abrading speed had high-abrading efficiency (high-volume loss) compared to low-abrading speed in all abrading instruments used. The diamond vitrified wheels demonstrated higher volume loss for two zirconia ceramics than those of SiC and diamond sintered wheels. When the high-speed air-turbine instruments were used, the diamond points showed higher volume losses compared to the carbide burs for one ceramic and two zirconia ceramics with high-mechanical properties. The results of this study indicated that the machinability of dental ceramics depends on the mechanical and physical properties of dental ceramics and machining instruments.

Open image in new window

The abrading wheels show autogenous action of abrasive grains, in which ground abrasive grains drop out from the binder during abrasion, then the binder follow to wear out, subsequently new abrasive grains come out onto the instrument surface (autogenous action) and increase the grinding amount (volume loss) of grinding materials.

     

High‐resolution OLED display with remarkably low power consumption using blue/yellow tandem structure and RGBY subpixels

A top‐emission organic light‐emitting diode (OLED) with a microcavity structure combined with a blue/yellow tandem structure was developed. A high‐resolution active‐matrix OLED display with the world's lowest level of power consumption using the tandem OLED with red, green, blue, and yellow subpixels was fabricated.