Runx1 Messenger RNA Delivered by Polyplex Nanomicelles Alleviate Spinal Disc Hydration Loss in a Rat Disc Degeneration Model

Vertebral disc degenerative disease (DDD) affects millions of people worldwide and is a critical factor leading to low back and neck pain and consequent disability. Currently, no strategy has addressed curing DDD from fundamental aspects, because the pathological mechanism leading to DDD is still controversial. One possible mechanism points to the homeostatic status of extracellular matrix (ECM) anabolism, and catabolism in the disc may play a vital role in the disease’s progression. If the damaged disc receives an abundant amount of cartilage, anabolic factors may stimulate the residual cells in the damaged disc to secrete the ECM and mitigate the degeneration process. To examine this hypothesis, a cartilage anabolic factor, Runx1, was expressed by mRNA through a sophisticated polyamine-based PEG-polyplex nanomicelle delivery system in the damaged disc in a rat model. The mRNA medicine and polyamine carrier have favorable safety characteristics and biocompatibility for regenerative medicine. The endocytosis of mRNA-loaded polyplex nanomicelles in vitro, mRNA delivery efficacy, hydration content, disc shrinkage, and ECM in the disc in vivo were also examined. The data revealed that the mRNA-loaded polyplex nanomicelle was promptly engulfed by cellular late endosome, then spread into the cytosol homogeneously at a rate of less than 20 min post-administration of the mRNA medicine. The mRNA expression persisted for at least 6-days post-injection in vivo. Furthermore, the Runx1 mRNA delivered by polyplex nanomicelles increased hydration content by ≈43% in the punctured disc at 4-weeks post-injection (wpi) compared with naked Runx1 mRNA administration. Meanwhile, the disc space and ECM production were also significantly ameliorated in the polyplex nanomicelle group. This study demonstrated that anabolic factor administration by polyplex nanomicelle-protected mRNA medicine, such as Runx1, plays a key role in alleviating the progress of DDD, which is an imbalance scenario of disc metabolism. This platform could be further developed as a promising strategy applied to regenerative medicine.     

A novel method for determining vitamin B1 in a wide variety of foodstuffs with or without polyphenols

Certain foodstuffs exhibit matrix interference effects on the vitamin B₁ analysis prescribed in the official methods of the European Union, AOAC International, and Japan. In this study, we demonstrated that one of the problematic polyphenols in green tea or cocoa was tannin. For these matrices, thiamine was found to interact with tannin in the extraction step and was thus removed by filtration as a residue. To resolve the matrix interference, we proposed lowering the pH of the extraction solution by adding hydrochloric acid after the enzyme reaction. High precision (2–3% RSDr) and good recovery (98.3–103%) were obtained with reference materials using the proposed method. We also confirmed the equivalence of the obtained data from the proposed method and the Japanese official method for nutritional labeling. From these results, the method was found to be effective for vitamin B₁ analysis regardless of the presence of interference matrices.     

Topical Phenomena Observed in a Short-Gap Atmospheric Discharge Using Rotor and Post Electrodes

Correlation of light emission, discharge structure, waveform of the discharge current, electrode configuration, and electromagnetic radiation is examined with the intent of obtaining an effective means for preventing electromagnetic interference (EMI) due to a short-gap discharge. The electromagnetic radiation (EMR) level resulting from a current step which, in turn, was formed by a discrete movement of a cathode spot was clearly recognized. A combination of needle rotor and needle post gave the smallest electromagnetic radiation level in the experiments.     

Verification of the energy-saving effect of the district heating and cooling system—Simulation of an electric-driven heat pump system

A district heating and cooling system (DHC) is expected to be a promising energy-saving measure for high-density business areas in Japan. However, it has not been verified what advantages of the DHC are important for energy conservation. The clarification of this issue is supposed to contribute to improving the energy efficiency of the DHC.     

Highly dense protein layers confirmed by atomic force microscopy and quartz crystal microbalance

Protein adsorption on a gold surface is investigated by comparing the results of quartz crystal microbalance method and atomic force microscopy. The adsorption of streptavidin on functional gold surfaces is directly monitored by a quartz crystal microbalance, and confirmed by atomic force microscopy. For this investigation, a modified gold substrate is fabricated to obtain a topographic image of streptavidin molecules. Both methods show a correlation in terms of the highly dense protein single-layer formation, and the modified gold electrode shows a slightly denser protein layer formation because of the difference in substrate geometry as compared with that of a mica surface.     

Prediction method of critical power by film flow rate measurement and subchannel analysis

This paper presents a method that can estimate the critical power of boiling water reactors, BWRs, with regard to spacer geometry. The current experimental method for estimating the critical power for BWR design requires many trained experts and expensive facilities to conduct the experiments. In the present method, the liquid film flow rate of adiabatic gas‐liquid two‐phase flow and a subchannel analysis of the actual BWR flow condition are measured experimentally and analyzed. In the experiment, deposition enhancement coefficients of three spacer geometries—a ferrule, an egg‐crate, and a ferrule spacer with twisted tape (CYCLONE spacer)—were estimated by measuring the liquid film flow rate of air‐water two‐phase flow flowing up in a vertical square (4 × 4) rod bundle that simulated the rod bundle of a BWR. Using these coefficients, the critical powers for bundles using each type of spacer geometry were calculated in the subchannel analysis. This method was validated using previous critical power data in the actual BWR flow condition. The critical powers predicted by this method agreed well with those of the experimental data. The result confirmed the effectiveness of this experiment‐simulation combined method, as well as the advantage over current experimental methods in terms of human and facility costs. © 2005 Wiley Periodicals, Inc. Heat Trans Asian Res, 34(5): 309–323, 2005; Published online in Wiley InterScience ( www.interscience.wiley.com ). DOI 10.1002/htj.20069     

Interactive configuration through force analysis of GM1 pentasaccharide-Vibrio cholera toxin interaction

Understanding of the molecular relationships in carbohydrate-protein interactions provides useful information on biological processes in living organisms and is also helpful for development of potent biomedical agents. Herein, the interaction unbinding force between GM1 pentasaccharide and Vibrio cholera toxin (ctx) proteins was measured using atomic force microscopy (AFM), which enabled us to determine the interaction of ctx holotoxin (ctxAB) with GM1 and the interactive formation. First, the interaction force measured between A and B subunits (ctxA-ctxB) was 184.2 ± 4.5 pN, and the unbinding forces were evaluated to confirm the role of ctxA in ctxAB complex formation and were determined to be 443.7 ± 7.5 and 535.7 ± 25.9 pN for GM1-ctxB and GM1-ctxAB complexes, respectively. The force difference of ～90 pN between GM1-ctxB and GM1-ctxAB might be due to the formation of the cholera toxin complex. Importantly, from the analogue analyses, we understand how structural and binding positional differences in complex carbohydrates affect the interaction with protein and surmise that the GM1-ctxAB complex makes a "two-finger grip" formation through the conformational change of a flexible carbohydrate. In conclusion, using AFM force analysis, we successfully quantified and characterized the interactive configuration of carbohydrate-protein molecules.     

CuPc/PbZr0.2Ti0.8O3/(La,Ba)MnO3 field effect transistor heterojunction photomemory

We investigated a photoconductor(PC)/ferroelectric oxide(OF)/semiconductor oxide(SO) POS-FET structure photomemory consisting of organic photoconductor CuPc/inorganic ferroelectric PbZr_0.2Ti_0.8O₃ heterojunction gate and a ferromagnetic oxide semiconductor La_0.87Ba_0.13MnO₃ channel. Visible light information detected by photoconductor CuPc can be memorized in ferroelectric PbZr_0.2Ti_0.8O₃, and non-volatile and non-destructive reading out process of light information memorized in this ferroelectric layer were achieved by reading out the resistance modulation of ferromagnetic semiconductor oxide La_0.87Ba_0.13MnO₃ channel under the photoconductor/ferroelectric gate.