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.     

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.     

Computer simulation of hydraulic fractures

We provide a brief historical background of the development of hydraulic fracturing models for use in the petroleum and other industries. We discuss scaling laws and the propagation regimes that control the growth of hydraulic fractures from the laboratory to the field scale. We introduce the mathematical equations and boundary conditions that govern the hydraulic fracturing process, and discuss numerical implementation issues including: tracking of the fracture footprint, the control of the growth of the hydraulic fracture as a function of time, coupling of the equations, and time-stepping schemes. We demonstrate the complexity of hydraulic fracturing by means of an application example based on real data. Finally, we highlight some key areas of research that need to be addressed in order to improve current models.     

Effect of Sliding History on Super-Low Friction of Diamond-Like Carbon Coating in Water Lubrication

Water lubrication using diamond-like carbon (DLC) coatings is regarded as being a promising eco-friendly technology. However, it has been reported that delamination of DLC coatings occurs in water. The authors clarified that the application of friction in ambient air prior to self-mating DLC water lubrication, in addition to the suppression of DLC delamination, caused the friction coefficient to decrease more than when no processing is performed in the boundary lubrication region. When compared to the case when there was no pre-sliding, in terms of the wear scar, in this case, a much more hydrophilic and smoother surface was formed. Therefore, the friction-reducing process in water due to the presence of a sliding history (pre-sliding in ambient air) was clarified by an analysis of the DLC geometry, structure, chemical state and an in situ analysis that can measure the friction properties and the chemical state of DLC. We clarified that the process in which the oxidation and structural changes of the DLC proceed at the time of pre-sliding in ambient air using in situ Fourier transform infrared spectroscopy–attenuated total reflection. It was also revealed that the OH termination subsequently accelerated rapidly due to the friction in water after pre-sliding process. In addition, a time-of-flight secondary ion mass spectrometry analysis revealed that this OH group was derived from the lubricating water and unlike the case in which there was no pre-sliding in ambient air, the rate at which OH terminates increased considerably.     

A comparison of approaches for finding minimum identifying codes on graphs

In order to formulate mathematical conjectures likely to be true, a number of base cases must be determined. However, many combinatorial problems are NP-hard and the computational complexity makes this research approach difficult using a standard brute force approach on a typical computer. One sample problem explored is that of finding a minimum identifying code. To work around the computational issues, a variety of methods are explored and consist of a parallel computing approach using MATLAB, an adiabatic quantum optimization approach using a D-Wave quantum annealing processor, and lastly using satisfiability modulo theory (SMT) and corresponding SMT solvers. Each of these methods requires the problem to be formulated in a unique manner. In this paper, we address the challenges of computing solutions to this NP-hard problem with respect to each of these methods.     

Swelling Pressure of Wet Rice Grains Estimated from Distribution Coefficients of Saccharides

The apparent density and porosity of four rice cultivars, Koshihikari, Yumepirika, Habutaemochi, and Kitayukimochi, decreased and increased, respectively, with the increasing temperature in the range of 5 °C to 60 °C. Especially large changes in both apparent density and porosity were observed at 60 °C, which can be ascribed to the starch gelatinization of rice. The swelling pressures of wet rice grains were estimated to be 7.5, 6.3, 6.2, and 5.9 MPa at 25 °C for Koshihikari, Yumepirika, Habutaemochi, and Kitayukimochi, respectively, from the distribution coefficients of solutes with different molar volumes. Rice grains having higher amylose content exhibited a weak tendency toward higher swelling pressure. The distribution coefficients of fructose for the Koshihikari and Habutaemochi cultivars were larger at higher temperatures, but the temperature dependence of the swelling pressure was not significant for both cultivars. These results suggested that the increase in the distribution coefficient was caused by the gradual relaxation of the interior structure of rice with increasing temperature.     

Moisture profiles of wheat noodles containing hydroxypropylated tapioca starch

Wheat noodles were prepared using flour to which hydroxypropylated tapioca starch was added, and the effect of this addition on the moisture distribution within the noodles during cooking was examined using a digital image processing technique. The addition of the modified starch slightly increased the moisture content and narrowed the flat distribution near the noodle surface. The distribution features reflected the changes in the water absorption behaviour caused by the properties of the modified starch and the reduction in the gluten content. Addition of the modified starch lowered Young's modulus and the energy for 99% strain of the noodles in the texture analysis to, at maximum 35% and 65%, and decreased the breakability of the noodles. These changes in the moisture distribution and textural properties have been ascribed to changes in both the state of the starch granules near the surface and the structure of the gluten network.