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.     

Voltage Control of Static Var Compensator for a Remote System Interconnected by Long‐Distance AC Cables

The voltage variation in a remote system is large when the system is connected by long‐distance AC cables due to the cable capacitance. In Japan, the longest 54‐km 66‐kV AC submarine cable interconnection between the Kyushu mainland and Goto Islands was commissioned in 2005. It was requested to mitigate the voltage variation caused by switching off and on one circuit of the two circuits in the AC cables when a fault occurs. Since the conventional voltage control methods such as transformer tap changer or shunt capacitor and reactor banks are not sufficient because of their slow response time, a static var compensator (SVC) was installed on the Goto Islands. In such an application, an SVC control method should be developed so as not to override the existing voltage control systems. This paper describes the SVC control method developed for the Goto Islands AC interconnection project, which can be applied to similar situations. The effectiveness of the control method was verified by the results of effective value simulation and of field testing, which was implemented before the SVC was commissioned in 2007. © 2013 Wiley Periodicals, Inc. Electr Eng Jpn, 186(3): 19–30, 2014; Published online in Wiley Online Library ( wileyonlinelibrary.com ). DOI 10.1002/eej.22337     

Dry season water allocation in the Chao Phraya River basin,Thailand

This study examines the recent quantitative characteristics of dry season water allocation in the Chao Phraya River basin, Thailand. Previous studies have focused on inequitable allocation, where the highest priority is given to the domestic water sector and uneven irrigation water variability exists among projects in the delta. This study uses a statistical test and panel data analysis to confirm that the characteristics highlighted in previous studies remain accurate, and it discusses the source of these characteristics in order to understand the issues in water allocation in the Chao Phraya River delta.     

Acceleration of the Rate of Silver Nanoparticle Formation Using Microbubbles in a Sonochemical Process

The effect of the addition of microbubbles on the formation of silver nanoparticles produced in an ultrasonic radiation-mediated process was investigated. The addition of microbubbles with an area-based median diameter of 26.0?µm and a bubble volume density of 0.18?cm³/L significantly increased the silver nanoparticle formation rate during the sonochemical process. The size distribution of the silver nanoparticles was largely unaffected by the addition of the microbubbles. The influence of changes in the microbubble volume density on the formation of the silver nanoparticles was also investigated; it was confirmed that the rate of formation of the silver nanoparticles increased moderately with increasing volume density. The gradient in absorption spectra was approximately 30 times larger for the case when microbubbles (median diameter: 28.3?µm) were added at 0.74?cm³/L, compared with the case without microbubbles. The results suggested that the microbubbles provided reaction sites similar to cavitation bubbles in the ultrasonic reaction.     

Catalytic cycle of cytochrome-c3 hydrogenase,a [NiFe]-enzyme,deduced from the structures of the enzyme and the enzyme mimic

Hydrogenases catalyze uptake and production of H₂. Heterolytic cleavage of H₂ bound on [NiFe]-hydrogenase (E) produces two unequal H species to form E:H_aH_b, where H_a and H_b behave differently. The structures of various states of the enzyme established by crystallography and spectroscopy were used to construct a catalytic cycle of the enzyme. The Ni–Fe center of the active enzyme has the Ni–Fe bridging site vacant. The enzyme is suggested to bind H₂ either at Ni or Fe atom. In E:H_aH_b, H_a is considered to be a protein-bound hydron (proton or deuteron) at the entrance to the hydrophobic gas tunnel. The structure of a synthetic hydrogenase-mimic suggests H_b to be the 6th ligand to Fe. Two successive one-electron processes from E:H_aH_b complete the catalytic cycle of H₂ uptake. The reverse of the cycle operates in the H₂ production. The proposed catalytic cycle is consistent with the kinetic, crystallographic and spectroscopic studies.     

Optimal operation of cogeneration system coordinated by aggregator for providing the demand and supply regulation capacity

Recent growth of renewable energy generations with natural variability, such as photovoltaic generation and wind turbine generation, would make the demand and supply control in a whole power system more difficult, and therefore, alternatives for demand and supply regulation resources would be required. The authors focus on cogeneration system (CGS) as one of regulation resources. In order to procure adequate volume of regulation capability, an aggregator coordinates a number of CGSs efficiently and flexibly considering the wide variety of electricity/thermal demands of CGS owners. This paper proposes a novel optimal operation strategy of CGS coordinated by the aggregator considering the energy balance and operation cost of individual CGS owner. This paper also demonstrates the availability of CGSs for regulation capability by numerical case studies in which the actual consumption profile is employed.     

Single-particle-diagnosis approach: An efficient strategy for discovering new nitride phosphors

Discovery of novel phosphors is extremely important to target the ever change of the solid state lighting technology. In this paper, we present a high-speed way to discover new phosphors, namely single-particle-diagnosis approach. This approach is based on investigating the crystal structure and luminescence of a tiny single crystal, without the necessity of the growth of large-size single crystals or the preparation of single phase powder samples. The concept of the approach and some new nitride phosphors explored by this approach are shown in this paper.     

Hydrophobic Tagged Dihydrofolate Reductase for Creating Misfolded Glycoprotein Mimetics

In the endoplasmic reticulum (ER), nascent glycoproteins that have not acquired the native conformation are either repaired or sorted for degradation by specific quality‐control systems composed by various proteins. Among them, UDP‐glucose:glycoprotein glucosyltransferase (UGGT) serves as a folding sensor in the ER. However, the molecular mechanism of its recognition remains obscure. This study used pseudo‐misfolded glycoproteins, comprising a modified dihydrofolate reductase with artificial pyrene–cysteine moiety on the protein surface (pDHFR) and Man₉GlcNAc₂‐methotrexate (M9‐MTX). All five M9‐MTX/pDHFR complexes, with a pyrene group at different positions, were found to be good substrates of UGGT, irrespective of the site of pyrene modification. These results suggest UGGT's mode of substrate recognition is fuzzy, thus allowing various glycoproteins to be accommodated in the folding cycle.     

Sieving of aerosol particles with metal screens

Metal screens with uniform micrometer-sized opening were employed to sieve aerosol particles by suppressing the adhesion of particles smaller than the openings. The collection efficiencies of monodispersed polystyrene latex (PSL) particles were experimentally determined using the metal screens with 1.2, 1.8, 2.5, and 4.2 μm openings at various filtration velocities. The particles smaller than the mesh opening adhered on the metal screen at a low filtration velocity, but the bounce-off of particles on the mesh surface suppressed the adhesion at a high velocity. As a result, we found that the adhesion of PSL particles larger than 0.3 μm mostly suppressed at a filtration velocity higher than 10 m s^?1 and therefore we can sieve aerosol particles according to the opening size of metal screens. We also found that the particle number concentration could be determined by measuring the increase in pressure drop since the clogging of metal screen openings takes place by the individual particles.

© 2016 American Association for Aerosol Research