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.     

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.     

Synthesis,aggregation and adsorption behaviour of a thermoresponsive pentablock copolymer

Poly(ethylene oxide)–poly(propylene oxide)–poly(ethylene oxide) (PEO–PPO–PEO) triblock copolymer (Pluronic F127) was modified by introducing poly(N‐isopropylacrylamide) (PNIPAM) at both the PEO ends, and the pentablock copolymer (PNIPAM₄₁–F127–PNIPAM₄₁, PN41) so prepared was characterized using gel permeation chromatography and ¹H NMR spectroscopy. The degree of polymerization of NIPAM blocks at the two ends was 41. The solution behaviour and microstructure of PN41 aggregates in water were examined using UV–visible spectroscopy, micro‐differential scanning calorimetry and small‐angle neutron scattering (SANS) and compared with F127. Two lower critical solution temperatures (LCSTs) were observed for the pentablock copolymer, corresponding to PPO and PNIPAM blocks, respectively. The adsorption of PN41 on thiol‐grafted hydrophobic gold surfaces at various temperatures was investigated using a quartz crystal microbalance. It was found that the adsorption behaviour and mechanism of PN41 were mainly determined by the interactions of the pentablock copolymers with different chain conformations in dilute aqueous solutions at various temperatures. SANS measurements were used to determine the temperature‐dependent structural evolution of polymer micelles in aqueous solution. A NOESY study revealed that above the LSCT of PNIPAM, the interaction of PPO and PNIPAM protons increases and the distance between PPO and PNIPAM decreases. © 2019 Society of Chemical Industry     

Blue Color Formation of Cyanobacteria with β-Cyclocitral

Volatile compounds, such as β-cyclocitral, geosmin, and 2-methylisoborneol, from cyanobacteria showed a lytic activity against cyanobacteria. Particularly, β-cyclocitral caused an interesting color change in the culture broth from green to blue during the lysis process. In the present study, the lytic behavior of various cyanobacteria with β-cyclocitral was investigated, and a mechanism for the blue color formation was developed. β-Cyclocitral lysed both the laboratory strains of any genera and bloom samples including many species of cyanobacteria, and caused the characteristic color change from green to blue. β-Cyclocitral provided a characteristic behavior, such that the absorption maxima of chlorophyll-a and β-carotene disappeared, but that of phycocyanin still remained after 12 h, which indicated that β-cyclocitral decomposed chlorophyll-a and β-carotene rapidly, so that the inherent colors from the tolerant water-soluble pigments became observable in the cultured broth. This phenomenon was confirmed by another experiment using Phormidium (NIES-611), which showed a pink color derived from phycoerythrin. β-Cyclocitral was more easily oxidized when compared with similar aldehyde compounds, so that the pH of the solution quickly decreased to 4.5. An oxidation product of β-cyclocitral in water solution was isolated and identified as 2,6,6-trimethylcyclohexene-1-carboxylic acid. This study provides support that β-cyclocitral derived from cyanobacteria plays an important role in the lysis of cyanobacteria and participates in the blue color formation under natural conditions.     

Electrochemical oxidation of cholesterol in acetonitrile leads to the formation of cholesta-4,6-dien-3-one

Cholesterol was shown to be oxidized at the glassy carbon electrode in an acetonitrile–2-propanol mixture and this oxidation reaction was applied to the determination of serum total cholesterol by high-performance liquid chromatography with electrochemical detection (K. Hojo, H. Hakamata, A. Ito, A. Kotani, C. Furukawa, Y.Y. Hosokawa, F. Kusu, J. Chromatogr. A 1166 (2007) 135–141). To gain insight into the detection mechanisms of cholesterol, an electrolytic product of cholesterol was collected and characterized by infrared spectroscopy, one- and two-dimensional nuclear magnetic resonance, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. The three techniques, together with comparisons of literature spectral data, confirmed the formation of cholesta-4,6-dien-3-one. The conversion of cholesterol to cholesta-4,6-dien-3-one, a four-electron, four-proton electrochemical process, has been proposed as an electrochemical oxidation mechanism of cholesterol in acetonitrile.     

The new and improved NIST Dragon's LAIR (Lofting and Ignition Research) facility

Several studies suggest that the firebrands are a major cause of structural ignition of Wildland–Urban Interface fires in USA and Australia. For 40 years, past firebrand studies have focused on how far firebrands fly and do not assess the vulnerabilities of structures to ignition from firebrand showers. The development of the National Institute of Standards and Technology (NIST) Dragon has allowed the quantification of structure ignition vulnerabilities of full‐scale building assemblies. Full‐scale tests are necessary to highlight vulnerabilities of structures to ignition under firebrand attack, whereas bench‐scale test methods afford the capability to test new firebrand‐resistant technologies and may serve as the basis for new standard testing methodologies. To this end, the present investigation was undertaken to construct a new and improved Dragon's Lofting and Ignition Research facility. This entailed removing the NIST Baby Dragon from the wind tunnel facility and inserting the new and improved NIST continuous feed Baby Dragon. The unique feature of the continuous feed Baby Dragon, over the current NIST Baby Dragon, is the capability to produce a constant firebrand shower in order to expose building materials to continual firebrand bombardment. The efficacy of the new experimental facility to determine ignition regime maps of building materials exposed to wind‐driven firebrand showers is presented. Specifically, ignition regime maps are presented as a function of continuous firebrand generation rate, wind tunnel speed, and cedar moisture content. Copyright © 2011 John Wiley & Sons, Ltd.     

Analgesic Mechanisms of Steroid Ointment against Oral Ulcerative Mucositis in a Rat Model

Despite the long history of use of steroid ointments for oral mucositis, the analgesic mechanism has not been fully elucidated. In this study, we examined the effects of triamcinolone acetonide (Tmc) on oral ulcerative mucositis-induced pain in conscious rats by our proprietary assay system. Based on evaluations of the physical properties and retention periods in the oral mucosa of human volunteers and rats, we selected TRAFUL^® ointment as a long-lasting base. In oral ulcerative mucositis model rats, TRAFUL^® with Tmc suppressed cyclooxygenase-dependent inflammatory responses with upregulations of glucocorticoid receptor-induced anti-inflammatory genes and inhibited spontaneous nociceptive behavior. When an ointment with a shorter residual period was used, the effects of Tmc were not elicited or were induced to a lesser extent. Importantly, TRAFUL^® with Tmc also improved oral ulcerative mucositis-induced mechanical allodynia, which has been reported to be independent of cyclooxygenase. Ca²⁺ imaging in dissociated trigeminal ganglion neurons showed that long-term preincubation with Tmc inhibited the hypertonic stimulation-induced Ca²⁺ response. These results suggest that the representative steroid Tmc suppresses oral ulcerative mucositis-induced pain by general anti-inflammatory actions and inhibits mechanical sensitivity in peripheral nerves. For drug delivery, long-lasting ointments such as TRAFUL^® are needed to sufficiently induce the therapeutic effects.     

Medical image diagnosis of kidney regions by deep feedback GMDH-type neural network using principal component-regression analysis

The deep feedback Group Method of Data Handling (GMDH)-type neural network is applied to the medical image recognition of kidney regions. In this algorithm, the principal component-regression analysis is used for the learning calculation of the neural network, and the accurate and stable predicted values are obtained. The neural network architecture is automatically organized so as to fit the complexity of the medical images using the prediction error criterion defined as Akaike’s Information Criterion (AIC) or Prediction Sum of Squares (PSS). The recognition results show that the deep feedback GMDH-type neural network algorithm is useful for the medical image recognition of kidney regions, because the optimum neural network architecture is automatically organized.