Split Enzyme-Based Biosensors for Structural Characterization of Soluble and Insoluble β-Glucans

β-Glucan is widely distributed in various plants and microorganisms and is composed of β-1,3-linked d-glucose units. It may have a branched short or long side chain of glucose units with β-1,6- or β-1,4-linkage. Numerous studies have investigated different β-glucans and revealed their bioactivities. To understand the structure-function relationship of β-glucan, we constructed a split-luciferase complementation assay for the structural analysis of long-chain β-1,6-branched β-1,3-glucan. The N- and C-terminal fragments of luciferase from deep-sea shrimp were fused to insect-derived β-1,3-glucan recognition protein and fungal endo-β-1,6-glucanase (Neg1)-derived β-1,6-glucan recognition protein, respectively. In this approach, two β-glucan recognition proteins bound to β-glucan molecules come into close proximity, resulting in the assembly of the full-length reporter enzyme and induction of transient luciferase activity, indicative of the structure of β-glucan. To test the applicability of this assay, β-glucan and two β-glucan recognition proteins were mixed, resulting in an increase in the luminescence intensity in a β-1,3-glucan with a long polymer of β-1,6-glucan in a dose-dependent manner. This simple test also allows the monitoring of real-time changes in the side chain structure and serves as a convenient method to distinguish between β-1,3-glucan and long-chain β-1,6-branched β-1,3-glucan in various soluble and insoluble β-glucans.     

Process Intensification of Living Cationic Polymerization of Isobutylene by a Flow Reaction System

Increasing the reaction temperature of the living cationic polymerization of isobutylene is crucial for industrial production due to the cost of refrigeration. The reaction temperature increase was achieved with an accelerated reaction rate using a flow reaction system. The polymerization conditions, including the flow reactor design, were based on the results of kinetic studies. Utilizing a milli‐scale flow reactor, polyisobutylene, which has a narrow molecular weight distribution, was obtained within a considerably short residence time at a high temperature. Furthermore, it was confirmed that the value of M_w/M_n correlates with the product of the Reynolds number and the angle of collision.     

Adaptation to climate change and conservation of biodiversity using green infrastructure

In recent years, we have experienced mega‐flood disasters in Japan due to climate change. In the last century, we have been building disaster prevention infrastructure (artificial levees and dams, referred to as “grey infrastructure”) to protect human lives and assets from floods, but these hard protective measures will not function against mega‐floods. Moreover, in a drastically depopulating society such as that in Japan, farmland abandonment prevails, and it will be more difficult to maintain grey infrastructure with a limited tax income. In this study, we propose the introduction of green infrastructure (GI) as an adaptation strategy for climate change. If we can use abandoned farmlands as GI, they may function to reduce disaster risks and provide habitats for various organisms that are adapted to wetland environments. First, we present a conceptual framework for disaster prevention using a hybrid of GI and conventional grey infrastructure. In this combination, the fundamental GI, composed of forests and wetlands in the catchment (GI‐1) and additional multilevel GIs such as flood control basins that function when floodwater exceeds the planning level (GI‐2) are introduced. We evaluated the flood attenuation function (GI‐1) of the Kushiro Wetland using a hydrological model and developed a methodology for selecting suitable locations of GI‐2, considering flood risk, biodiversity and the distribution of abandoned farmlands, which represent social and economic costs. The results indicated that the Kushiro Wetland acts as a large natural reservoir that attenuates the hydrological peak discharge during floods and suitable locations for introducing GI‐2 are concentrated in floodplain areas developing in the downstream reaches of large rivers. Finally, we discussed the network structure of GI‐1 as a hub and GI‐2 as a dispersal site for conservation of the Red‐crowned Crane, one of the symbolic species of Japan.     

Quantitative Elemental Analysis of a Single Cell by Using Inductively Coupled Plasma-Mass Spectrometry in Fast Time-Resolved Analysis Mode

The elemental composition of a single yeast, green alga, or red blood cell (RBC) was precisely determined by using inductively coupled plasma-mass spectrometry (ICP-MS) operating in fast time-resolved analysis (TRA) mode. The technique is known as single-cell (SC)-ICP-MS. Phosphorus, sulfur, magnesium, zinc, and iron were detected in the three types of cell. The elemental composition of yeast and green alga obtained by SC-ICP-MS was consistent with results obtained from conventional ICP-MS measurements following acid digestion of the cells. Slight differences were found in the measured values between SC-ICP-MS and the conventional ICP-MS results for RBC. However, the SC-ICP-MS results for S and Fe in RBC were closer to the estimated values for these elements that were calculated from the level of hemoglobin in RBCs. The data suggest that SC-ICP-MS is suitable for the analysis of various cell types, namely, fungus, plant, and animal cells.     

First nuclear transmutation of 237Np and 241Am by accelerator-driven system at Kyoto University Critical Assembly

This study demonstrates, for the first time, the principle of nuclear transmutation of minor actinide (MA) by the accelerator-driven system (ADS) through the injection of high-energy neutrons into the subcritical core at the Kyoto University Critical Assembly. The main objective of the experiments is to confirm fission reactions of neptunium-237 (²³⁷Np) and americium-241 (²⁴¹Am), and capture reactions of ²³⁷Np. Subcritical irradiation of ²³⁷Np and ²⁴¹Am foils is conducted in a hard spectrum core with the use of the back-to-back fission chamber that obtains simultaneously two signals from specially installed test (²³⁷Np or ²⁴¹Am) and reference (uranium-235) foils. The first nuclear transmutation of ²³⁷Np and ²⁴¹Am by ADS soundly implemented by combining the subcritical core and the 100 MeV proton accelerator, and the use of a lead-bismuth target, is conclusively demonstrated through the experimental results of fission and capture reaction events.     

Possibility of Japanese Cedar Pollen Causing False Positives in the Deep Mycosis Test

Because Japanese cedar pollen (JCP) contains beta-1,3-d-glucan (BG), there is concern that its lingering presence in the atmosphere, especially during its scattering period, may cause false positives in the factor-G-based Limulus amebocyte lysate (LAL) assay used to test for deep mycosis (i.e., G-test). Hence, we examined whether the LAL assay would react positively with substances contained in JCP by using the G-test to measure JCP particles and extracts. BG was purified from the JCP extract on a BG-specific affinity column, and the percentage extractability was measured using three different BG-specific quantitative methods. The G-test detected 0.4 pg BG in a single JCP particle and 10 fg from a single particle in the extract. The percentage extractability of JCP-derived BG was not significantly different among the three quantitative methods. As the JCP particles should technically have been removed during serum separation, they should be less likely to be a direct false-positive factor. However, given that the LAL-assay-positive substances in the JCP extract were not distinguishable by the three BG-specific quantitative methods, we conclude that they may cause the background to rise. Therefore, in Japan false positives arising from JCP contamination should be considered when testing patients for deep mycosis.     

Production of thin graphite sheets for a high electrical conductivity film by the mechanical delamination of ternary graphite intercalation compounds

Herein we propose a production scheme for conductive films composed of thin graphite sheets with high crystallinity and polymeric resin. The crystalline graphite sheets were successfully produced from natural graphite powder by solution-phase synthesis of graphite intercalation compounds (GICs), following a wet planetary-ball milling under mild conditions. The shear forces in the milling pot lead to a peeling of graphite flakes. Taking into consideration the interlayer bonding force, the delamination should be preferentially done from the expanded GICs interlayer rather than intrinsic graphite one. Some composite films derived from the phenolic resin and flaky graphite sheets displayed much higher electrical conductivities compared to the film from the feed graphite particles. We also demonstrate the stage structure of synthetic GICs affected the film conductivity. The composite films made from exfoliated products of ground (around stage IV) GICs exhibited high electrical conductivity with a small amount of the graphite sheets.     

Diphenyl Carbonate Synthesis by Homogeneous Pd Electrocatalyst

Electrochemical carbonylation of phenol with CO to diphenyl carbonate (DPC) using Pd electrocatalyst was studied at P(CO) = 1 atm and room temperature. Electrochemical carbonylation was conducted by galvanostatic electrolysis at 1 mA in electrolyte solutions containing phenol, sodium phenoxide and CH₃CN. It was found that electrochemical carbonylation by Pd was promoted by using 1,3-dimesitylimidazol-2-ylidene (IMes) ligand of a N-heterocyclic carbene. The current efficiency of DPC increased more than 3.4 times using the IMes ligand. The maximum current efficiency of DPC formation was 45 %.     

Effect of Vacancy Distribution on the Thermal Conductivity of Ga<Subscript>2</Subscript>Te<Subscript>3</Subscript> and Ga<Subscript>2</Subscript>Se<Subscript>3</Subscript>

Our group has focused attention on Ga₂Te₃ as a natural nanostructured thermoelectric material. Ga₂Te₃ has basically a zincblende structure, but one-third of the Ga sites are structural vacancies due to the valence mismatch between Ga and Te. It has been confirmed that (1) vacancies in Ga₂Te₃ exist as two-dimensional (2D) vacancy planes, and (2) Ga₂Te₃ exhibits an unexpectedly low thermal conductivity (κ), most likely due to highly effective phonon scattering by the 2D vacancy planes. However, the effect of the size and periodicity of the 2D vacancy planes on κ has been unclear. In addition, it has also been unclear whether only the 2D vacancy planes reduce κ or if point-type vacancies can also reduce κ. In the present study, we tried to prepare Ga₂Te₃ and Ga₂Se₃ with various vacancy distributions by controlling annealing conditions. The atomic structures of the samples were characterized by means of transmission electron microscopy, and κ was evaluated from the thermal diffusivity measured by the laser flash method. The effects of vacancy distributions on κ of Ga₂Te₃ and Ga₂Se₃ are discussed.     

Effects of Tl-filling into the voids and Rh substitution for Co on the thermoelectric properties of CoSb3

In order to enhance the thermoelectric (TE) properties of CoSb₃, we tried to reduce the lattice thermal conductivity (κ_lat) by filling Tl into the voids and substitution of Rh for Co. We prepared polycrystalline samples of Tl_x(Co_1−yRh_y)₄Sb₁₂ (x = 0, 0.05, 0.10, 0.15, 0.20 and y = 0.1, 0.2) and examined their TE properties from room temperature to 750 K. All the samples indicated negative values of the Seebeck coefficient (S). Both the electrical resistivity and the absolute values of the S decreased with increasing the Tl-filling ratio. The Tl-filling and Rh substitution reduced the κ_lat, due to the rattling and the alloy scattering effects. The minimum value of the κ_lat was 1.54 W m⁻¹ K⁻¹ at 550 K obtained for Tl_0.20(Co_0.8Rh_0.2)₄Sb₁₂. Tl_0.20(Co_0.8Rh_0.2)₄Sb₁₂ exhibited the best TE performance; the maximum value for the dimensionless figure of merit ZT was 0.58 at around 600 K.