Comparison of isolation methods using commercially available kits for obtaining extracellular vesicles from cow milk

Extracellular vesicles (EV) are important for delivering biologically active substances to facilitate cell-to-cell communication. Milk-derived EV are widely known because of their potential for immune enhancement. However, procedures for isolating milk-derived EV have not been fully established. To obtain pure milk-derived EV and accurately reveal their function, such procedures must be established. The aim of the present study was to compare methods using commercially available kits for isolating milk-derived EV. Initially, we investigated procedures to remove casein, which is the major obstacle in determining milk-derived EV purity. We separated whey using centrifugation only, acetic acid precipitation, and EDTA precipitation. Then, we isolated milk-derived EV by ultracentrifugation, membrane affinity column, size exclusion chromatography (SEC), polymer-based isolation, or phosphatidylserine-affinity isolation. Using EV count per milligram of protein, which is a good indicator of purity, we determined that acetic acid precipitation was the best method for removing casein. Using nanoparticle tracking analysis, protein quantity analysis, and RNA quantity analysis, we comprehensively compared each isolation method for its purity and yield. We found that SEC-based qEV column (Izon Science) could collect purer milk-derived EV at higher quantities. Thus, a combination of acetic acid precipitation and qEV can effectively isolate high amounts of pure extracellular vesicles from bovine milk.     

Syndiotactic‐specific radical polymerization of N‐isopropylacrylamide in toluene at low temperatures in the presence of silyl alcohols

The radical polymerization of N‐isopropylacrylamide was carried out in toluene at low temperatures in the presence of silyl alcohols, such as triethylsilanol. Poly(N‐isopropylacrylamide) with a racemo dyad content of 75% was obtained at ? 80 °C with a 4:1 triethylsilanol to monomer ratio loading. NMR analysis suggests that the mechanism for syndiotactic induction, in the presence of silyl alcohols, may be similar to that observed with alkyl alcohols. In this case, a 1:2 complex formation, via hydrogen bonding interactions, leads to the induction of syndiotactic specificity. Copyright © 2012 Society of Chemical Industry     

Evaluating the protective effects of radon inhalation or ascorbic acid treatment after transient global cerebral ischemic injury in gerbils

In this study, we compared the protective effects of radon inhalation and ascorbic acid administration on transient global cerebral ischemic injury in gerbils. Gerbils were treated with radon inhalation (2000 Bq/m³, 24 hours) or ascorbic acid (100, 300, or 500 mg/kg body weight). Then, transient global cerebral ischemia was induced by bilateral occlusion of the common carotid artery. Results showed that the number of damaged neurons was significantly increased in gerbils that underwent ischemia compared with that in control gerbils. However, the number of damaged neurons in gerbils treated with radon or 500 mg/kg of ascorbic acid before ischemia was significantly lower than gerbils who were subjected to ischemia without any pretreatment, and the protective effects of radon inhalation were similar to the effects of administering 500 mg/kg ascorbic acid. The levels of superoxide dismutase (SOD) and total glutathione (t-GSH) in brain tissue were increased to a similar extent by pretreatment with radon inhalation or 500 mg/kg of ascorbic acid. These findings suggested that radon inhalation has a protective antioxidative effect against transient global cerebral ischemic injury similar to 500 mg/kg ascorbic acid treatment.     

WO3/BiVO4 composite photoelectrode prepared by improved auto-combustion method for highly efficient water splitting

We report on the improvement in the water splitting efficiency of a WO₃/BiVO₄ composite photoelectrode by the application of an improved auto-combustion method to the preparation of porous BiVO₄ thin films. The unique feature of this preparation method is the addition of both NH₄NO₃, as a strong oxidizing agent, and an organic additive into BiVO₄ precursor solution. The local decomposition heat of the organic additive and oxidizing agent created a porous film with small, highly crystalline BiVO₄ particles. The photoelectrode has many advantages over existing ones, such as the high light-harvesting efficiency (LHE), a single BiVO₄ phase, the facile access of the holes to the photoelectrode/electrolyte interface, and the ease of water and oxygen diffusion. The maximum incident photon-to-current efficiency (IPCE) was estimated to be 64% (at 440 nm, 1.23 V vs. RHE) and the applied bias photon-tocurrent efficiency (ABPE) reached as high as 1.28%. This ABPE value is highest among all oxide semiconductor photoelectrodes reported previously, except for the case of a stacking photoelectrode system.     

Simulation of a chemical reaction, 2LiH→Li2+H2, driven by doubly excitation

A direct computer simulation of reaction dynamics at the electronic excited states is not easy to perform, because nonadiabatic equations must be solved as a function of time. Here we present a simple simulation to integrate directly the time-dependent Schrödinger equation within the framework of the time-dependent density functional theory (for electrons) coupled with the Newtonian equation of motion (for nuclei). We find that a chemical reaction, 2LiH→Li₂+H₂, takes place by the doubly excitation. Along the reaction, a level crossing occurs automatically between the highest occupied and lowest unoccupied levels. The simulation demonstrates a mechanism for relaxation for the reactions driven by doubly excitation: electronic excited state changes smoothly into the electronic ground state leaving a kinetic energy of the atoms.     

Robust comprehension of natural language instructions by a domestic service robot

We present a method through which domestic service robots can comprehend natural language instructions. For each action type, a variety of natural language expressions can be used, for example, the instruction, ‘Go to the kitchen’ can also be expressed as ‘Move to the kitchen.’ We are of the view that natural language instructions are intuitive and, therefore, constitute one of the most user-friendly robot instruction methods. In this paper, we propose a method that enables robots to comprehend instructions spoken by a human user in his/her natural language. The proposed method combines action-type classification, which is based on a support vector machine, and slot extraction, which is based on conditional random fields, both of which are required in order for a robot to execute an action. Further, by considering the co-occurrence relationship between the action type and the slots along with the speech recognition score, the proposed method can avoid degradation of the robot’s comprehension accuracy in noisy environments, where inaccurate speech recognition can be problematic. We conducted experiments using a Japanese instruction data-set collected using a questionnaire-based survey. Experimental results show that the robot’s comprehension accuracy is higher in a noisy environment using our method than when using a baseline method with only a 1-best speech recognition result.     

Numerical computation of two-loop box diagrams with masses

A new approach is presented to evaluate multi-loop integrals, which appear in the calculation of cross-sections in high-energy physics. It relies on a fully numerical method and is applicable to a wide class of integrals with various mass configurations. As an example, the computation of two-loop planar and non-planar box diagrams is shown. The results are confirmed by comparisons with other techniques, including the reduction method, and by a consistency check using the dispersion relation.     

Adhesion between polymer surface modified by graft polymerization and tissue during surgery using an ultrasonically activated scalpel device

In the field of surgery, achieving adhesion between a polymer implant and tissue poses a challenge considering that suturing is not appropriate for the stability of such implants. An ultrasonically activated scalpel that generates heat by mechanical vibration and promotes adhesion between a polymer implant and native tissue by pressing the two materials together has very good potential for application in the field. To determine the type of polymer that is suitable for the purpose, we investigated polyethylene (PE) and polystyrene (PS) films, the surfaces of which were activated by corona discharge. Graft polymerization was then performed on the corona‐treated surfaces to vary their properties. The corona‐treated PE and PS films grafted with poly(acrylic acid), poly(methacrylic acid), poly(vinyl benzylacrylic acid), and poly(hydroxylethyl acrylate), respectively, adhered to the tissue when the ultrasonically activated scalpel was applied. The heat generated by the mechanical vibration and the applied pressure enabled the carboxyl or hydroxyl groups to bond with the proteins in the extracellular matrix. We therefore concluded that it was possible to integrate this technique in the development of new types of polymer devices that could be stably implanted in a living body. © 2014 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2014 , 131, 40885.