Two Triterpene Synthases from Imperata cylindrica Catalyzing the Formation of a Pair of Diastereoisomers through Boat or Chair Cyclization

Imperata cylindrica is known to produce a pair of triterpenes, isoarborinol and fernenol, that exhibit identical planar structures but possess opposite stereochemistry at six of the nine chiral centers. These differences arise from a boat or a chair cyclization of the B-ring of the substrate. Herein, we report the characterization of three OSC genes from I. cylindrica. IcOSC1 and IcOSC5 were identified as isoarborinol and fernenol synthases, respectively, while IcOSC3 was characterized as a multifunctional enzyme that produces glutinol and friedelin as its major products. Mutational studies of isoarborinol and fernenol synthases revealed that the residues surrounding the DCTAE motif partially affected the conformation of the B-ring during cyclization. Additionally, the IcOSC1-W255H mutant produced the rare triterpene boehmerol. The introduced histidine residue presumably abstracted a proton from the intermediary carbocation at C18 during the 1,2-rearrangement. Expression analysis indicated that all OSC genes were highly expressed in stems.     

Developmental ability of cloned embryos from neural stem cells

The success rate is generally higher when cloning mice from embryonic stem (ES) cell nuclei than from somatic cell nuclei, suggesting that the embryonic nature or the undifferentiated state of the donor cell increases cloning efficiency. We assessed the developmental ability of cloned embryos derived from cultured neural stem cell (NSC) nuclei and compared the success rate with that of embryos cloned from other donor cells such as differentiated NSCs, cumulus cells, Sertoli cells and ES cells in the mouse. The transfer of two-cell cloned embryos derived from cultured NSC nuclei into surrogate mothers produced five live cloned mice. However, the success rate (0.5%) was higher in embryos cloned from cultured NSC nuclei than from differentiated NSCs (0%), but lower than that obtained by cloning mice from other cell nuclei (2.2-3.5%). Although the in vitro developmental potential to the two-cell stage of the cloned embryos derived from NSC nuclei (73%) was similar to that of the cloned embryos derived from other somatic cell nuclei (e.g., 85% in Sertoli cells and 75% in cumulus cells), the developmental rate to the morula-blastocyst stage was only 7%. This rate is remarkably lower than that produced from other somatic cells (e.g., 50% in Sertoli cells and 54% in cumulus cells). These results indicate that the undifferentiated state of neural cells does not enhance the cloning efficiency in mice and that the arrest point for in vitro development of cloned embryos depends on the donor cell type.     

Molecular structures of gellan gum imaged with atomic force microscopy (AFM) in relation to the rheological behavior in aqueous systems in the presence of sodium chloride

Aqueous solutions of gellan gum with comparable molecular mass but with different acyl contents were investigated by atomic force microscopy and rheological measurements in the presence of sodium chloride (NaCl). Results obtained were discussed in relation to our previous report using potassium chloride (KCl) as an added salt. For a low-acyl sample, continuous fibrous network structures were identified microscopically as in the case of KCl. The network structures were more heterogeneous than those formed with KCl in terms of the height distribution of molecular assemblies. Rheological thermal hysteresis between sol–gel transitions was detected as in the case of KCl. The storage modulus (G′) of the gelled system was ca. 15% of the corresponding data with KCl at 20 °C. For a high-acyl sample, no continuous network structures were identified but branches with observable ends were identified as in the case of KCl. The hysteresis was less evident than the corresponding data with KCl and for the low-acyl sample with NaCl. Also, G′ values at 20 °C were ca. 30% and 20% of the corresponding data with KCl and for the low-acyl sample with NaCl, respectively. Continuousness and homogeneity of network structures related to the hysteresis and elasticity of the system, respectively.     

Understanding structure ignition vulnerabilities using mock‐up sections of attached wood fencing assemblies

Firebrand production from structure combustion becomes a key factor in the magnitude of how quickly a large outdoor fire may spread. Post‐fire disaster investigations suggest that attached building components, such as wood fencing assemblies are known to be prone to ignition in these fires, and may provide pathways to structure ignition. Here, a comparison of ignition results from full‐scale fencing assembly experiments conducted using a full‐scale wind tunnel facility, to mock‐ups of full‐scale fencing assemblies using the recently developed experimental capability at the National Research Institute of Fire and Disaster (NRIFD) are discussed. In both experimental facilities, the fencing assemblies were exposed to firebrand showers using custom built continuous feed firebrand generators with size and mass distributions similar to those generated from structure combustion. Similar ignition behaviors were observed between the full‐scale fencing assemblies and the mock‐up of full‐scale fencing assemblies. Additional experiments are required for other fencing assembly types to further verify these important findings.     

Carbon radicals generated by solid polymers: Electron spin resonance spectroscopy for detection of species in water

Radicals generated in plastic medical devices (solid phase) by γ-rays or electron-beam irradiation during sterilization are known to cause oxidation of protein drugs, resulting in a loss or reduction in drug efficacy. The generation of radical species in water by the radical species in solid polymers has not been proved. Using electron spin resonance (ESR) spectroscopy, we confirm the generation of new radicals in water by γ-ray irradiated cyclic olefin polymers (COP). ESR measurements are obtained using 4-hydroxy-2,2,6,6-tetramethylpiperidine-1-oxyl (TEMPOL) as a spin probe and 5-(2,2-dimethyl-1,3-propoxycyclophosphoryl)-5-methyl-1-pyrroline N-oxide (CYPMPO) as a spin trap, in which the irradiated COP was immersed. The ESR signals indicate the TEMPOL radicals decline over time, suggesting the generation of new radicals. Conversely, the characteristic ESR signals of the adduct formed by the reaction between CYPMPO and the hydroxyl radical are observed. Thus, hydroxyl radicals are generated because of the migration of the radicals from COP to water. © 2019 Wiley Periodicals, Inc. J. Appl. Polym. Sci. 2020 , 137, 48604.     

An efficient manufacturing method for I-shaped cross-sectional CFRP beam with arbitrary arrangement of carbon fiber using electro-activated resin molding

Abstract

The I-shaped cross-sectional beam of CFRP (CFRP I-beam) is usually manufactured by the continuous protrusion method. Carbon fibers can only be arranged in the longitudinal direction. The CFRP I-beam with arbitrary arrangement of carbon fiber was manufactured with applying the electro-activated deposition molding method. The carbon fiber fabric was immersed in the deposition solution and energized, epoxy resin precipitated around carbon fiber and impregnated. The resin-impregnated fabric was installed to the mold, and the CFRP I-beam was fabricated. The CFRP I-beam was subjected to three-point bending tests, and the relationship between load-deflection was simulated by finite-element analysis.     

Experimental investigation of structure vulnerabilities to firebrand showers

Attempting to experimentally quantify the vulnerabilities of structures to ignition from firebrand showers has remained elusive. The coupling of two facilities has begun to unravel this difficult problem. The NIST Firebrand Generator (NIST Dragon) is an experimental device that can generate a firebrand shower in a safe and repeatable fashion. Since wind plays a critical role in the spread of WUI fires in the USA and urban fires in Japan, NIST has established collaboration with the Building Research Institute (BRI) in Japan. BRI maintains one of the only full scale wind tunnel facilities in the world designed specifically for fire experimentation; the Fire Research Wind Tunnel Facility (FRWTF). The present investigation is aimed at extensively quantifying firebrand penetration through building vents using full scale tests. A structure was placed inside the FRWTF and firebrand showers were directed at the structure using the NIST Dragon. The structure was fitted with a generic building vent, consisting of only a frame fitted with a metal mesh. Six different mesh sizes openings were used for testing. Behind the mesh, four different materials were placed to ascertain whether the firebrands that were able to penetrate the building mesh assembly could ignite these materials. Reduced scale test methods afford the capability to test new vent technologies and may serve as the basis for new standard testing methodologies. As a result, a new experimental facility developed at NIST is presented and is known as the NIST Dragon's LAIR (Lofting and Ignition Research). The NIST Dragon's LAIR has been developed to simulate a wind driven firebrand attack at reduced scale. The facility consists of a reduced scale Firebrand Generator (Baby Dragon) coupled to a bench scale wind tunnel. Finally, a series of full scale experiments were conducted to visualize the flow of firebrands around obstacles placed downstream of the NIST Dragon. Firebrands were observed to accumulate in front of these obstacles at a stagnation plane, as was observed when the structure was used for firebrand penetration through building vent experiments, due to flow recirculation. The accumulation of firebrands at a stagnation plane presents a severe threat to ignitable materials placed near structures.     

Analysis of molecular dynamics of moist wood components by applying the stretched-exponential function

On the basis of the molecular dynamics of wood components, the effects of adsorption water molecules on the change of relaxation behavior were examined for wood specimens with various moisture contents using the low of mixture and stretched-exponential function (so-called the Kohlrausch–Williams–Watts (KWW) function). The KWW function has two parameters: the characteristic relaxation time and a parameter related to the dispersion of the relaxation time. Both parameters and the relaxation modulus at time 10 s, E(10), could be separated into three regions corresponding to moisture contents of 0–0.05, 0.05–0.10, and greater than 0.10. Experimental results provided a consistent explanation for the relationships between the isotherm curve analysis of wood, the dielectric relaxation behavior of adsorption water molecules, and the mechanical relaxation behavior of wood.     

Effects of siRNAs targeting the spacer sequence of plant RNAi vectors on the specificity and efficiency of RNAi

Small interfering RNAs (siRNAs) generated from the spacer region of hairpin RNAs were not detected in the RNA interference (RNAi) plants targeting the fatty acid desaturase gene. The expression of the desaturase gene was stably suppressed even when siRNAs targeting the spacer sequences were introduced into this plant.