Isolation and Characterization of Two Types of Xyloglucanases from a Phytopathogenic Fungus,Verticillium dahliae

Xyloglucan is a major hemicellulosic component in plant cell walls. Phytopathogenic fungi secrete cell wall-degrading enzymes on their infection to hosts, while the nature of the cell wall-lytic enzymes of such fungi are yet to be fully understood. Verticillium dahliae is a soil-borne fungus that causes vascular wilt diseases in a variety of commercially important crops worldwide. We purified two types of xyloglucanases, XEG12A and XEG74B, from the culture of naturally isolated Verticillium dahliae strain 2148. XEG12A showed a molecular size of 23 kDa with its maximal activity at pH 7.5. XEG12A specifically hydrolyzed xyloglucan with no activity on other β-glucans. XEG74B had a molecular size of 110 kDa with its optimum pH at 6.0. XEG74B primarily hydrolyzed xyloglucan, with a slight activity on β-1,3-1,4-glucan. Analysis of hydrolytic products of xyloglucanooligasaccharide (XXXGXXXG) by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) revealed that the both enzymes cleaved β-1,4-glucosidic linkage at the position of unbranched chain, while XEG74B showed a little fluctuation with the cleavage site. Both enzymes did not hydrolyzed xyloglucanoheptasaccharide (XXXG) at all. N-Terminal and internal amino acid sequencing of the enzymes revealed that XEG12A and XEG74B belonged to Glycoside Hydrolase (GH) Families 12 and 74, respectively. Based on these results we concluded that V. dahliae XEG12A and XEG74B were xyloglucan-specific endo-β-1,4-glucanases (EC 3.2.1.151).     

Enhanced catalytic activity of nanoshell carbon co-doped with boron and nitrogen in the oxygen reduction reaction

The use of carbon cathode catalysts in polymer electrolyte fuel cells instead of the current platinum catalysts is attracting increasing attention. We claim that two factors are important for enhancing the activity of carbon cathode catalysts in the oxygen reduction reaction (ORR): the formation of a nanoshell structure and co-doping with boron and nitrogen. Herein, we investigate the preparation and characterization of active ORR carbon catalysts that combine the above factors. Boron and nitrogen (BN)-doped nanoshell-containing carbon (BN-NSCC) was prepared by carbonizing a mixture of poly(furfuryl alcohol), cobalt phthalocyanine, melamine, and a trifluoroborane–methanol complex at 1000 °C. Transmission electron microscopy and X-ray photoelectron spectroscopy revealed the formation of nanoshell structures with distorted graphitic layers and the introduction of boron and nitrogen atoms, respectively. The ORR activity was evaluated in oxygen-saturated 0.5 mol dm^?3 H₂SO₄ using Koutecky–Levich analysis. The BN-NSCC showed an eight to ten times higher ORR activity than undoped NSCC, with an increased number of electrons participating in the reaction. Tafel analysis revealed a change in the rate-determining step caused by BN-doping. Thus, the combination of a nanoshell structure and co-doping with boron and nitrogen was found to improve the ORR activity of carbon catalysts.     

Grain Boundary Segregation Behavior of Boron in Low-Alloy Steel

The boron concentration profiles around prior austenite grain boundaries in Fe-0.05C-0.5Mo-0.001B (mass pct) are examined using aberration-corrected STEM-EELS. In order to obtain the precise distribution of boron around the boundaries, tilt series measurements with thin specimens (<30 nm) are performed and the EEL spectra are analyzed by principal component analysis (PCA) and multivariate curve resolution (MCR). The boron concentration profile changes with the cooling rate from the solid solution temperature. The concentration at grain boundaries is maximized at a medium rate (30 °C/s), where the concentration reaches 8 at. pct, and it decreases at a larger (250 °C/s) or smaller (5 °C/s) rate. On the other hand, the boron distribution becomes wider as the cooling rate becomes smaller. The current results suggest that the boron segregation in the alloy is formed by the “non-equilibrium segregation mechanism.”     

Modeling of quantitative relationships between physicochemical properties of active pharmaceutical ingredients and tensile strength of tablets using a boosted tree

Objectives: The aim of this study was to explore the potential of boosted tree (BT) to develop a correlation model between active pharmaceutical ingredient (API) characteristics and a tensile strength (TS) of tablets as critical quality attributes.

Methods: First, we evaluated 81 kinds of API characteristics, such as particle size distribution, bulk density, tapped density, Hausner ratio, moisture content, elastic recovery, molecular weight, and partition coefficient. Next, we prepared tablets containing 50% API, 49% microcrystalline cellulose, and 1% magnesium stearate using direct compression at 6, 8, and 10?kN, and measured TS. Then, we applied BT to our dataset to develop a correlation model. Finally, the constructed BT model was validated using k-fold cross-validation.

Results: Results showed that the BT model achieved high-performance statistics, whereas multiple regression analysis resulted in poor estimations. Sensitivity analysis of the BT model revealed that diameter of powder particles at the 10th percentile of the cumulative percentage size distribution was the most crucial factor for TS. In addition, the influences of moisture content, partition coefficients, and modal diameter were appreciably meaningful factors.

Conclusions: This study demonstrates that BT model could provide comprehensive understanding of the latent structure underlying APIs and TS of tablets.     

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.     

Local structure and photocatalytic property of sol–gel synthesized ZnO doped with transition metal oxides

ZnO nanoparticles doped with up to 5 at% of Co and Mn were prepared using a co-precipitation method. The location of dopant ions and the effect of doping on the photocatalytic activity were investigated. The crystal structure of nanoparticles and local atomic arrangements around dopant ions were analyzed by X-ray absorption spectroscopy. The results showed that the Co ions substituted the Zn ions in the ZnO wurtzite phase structure and induced lattice shrinkage, while Mn ions were not completely incorporated in the crystal lattice. The photocatalytic activity under simulated sunlight was characterized by the decomposition of Rhodamine B dye molecules. It was revealed that Co-doping strongly reduced the photocatalytic activity but Mn-doping showed a weaker effect on the reduction of the photoactivity.     

Antitermitic activity of extracts from Chamaecyparis obtusa branch heartwood

The antitermitic activity of extracts from the branch heartwood of Chamaecyparis obtusa (hinoki) against Japanese termites (Reticulitermes speratus) was compared with that of the trunk. Samples of branch and trunk heartwood were extracted with n-hexane, ethyl acetate, and methanol successively. n-Hexane extracts of branch and trunk heartwood were strongly antitermitic, and branch heartwood contained greater quantities of active n-hexane extracts than trunk heartwood. Germacra-1-(10), 5-dien-4β-ol, t-cadinol, α-cadinol, hinokiresinol, and hinokinin were separated from the branch extracts and the termiticidal and antifeedant activity of these compounds was tested by no-choice and dual-choice test methods. The sesquiterpenoids, germacra-1-(10), 5-dien-4β-ol, t-cadinol and α-cadinol were strongly termiticidal. The norlignan hinokiresinol and lignan hinokinin had weak termiticidal, and strong antifeedant and repellent activity. High concentrations of germacra-1-(10), 5-dien-4β-ol and hinokiresinol were present in branch heartwood. These compounds protect hinoki branches from termites and other harmful organisms. Hinoki branch heartwood, which is usually unused, is a potential source of active antitermitic compounds.     

Connection‐Strength Estimation of Neuronal Networks by Fitting for Izhikevich Model

Recently, there has been abundant research using multineuron recording, but there are many problems with extracting the features from the obtained spike time series, which are huge in volume and complex. Here we introduce a new method of estimating synaptic connection strengths between neurons by fitting to the Izhikevich model by maximum likelihood estimation. We demonstrate that our method can estimate connection strengths from spike time series given by a simulated neural ensemble and can estimate nonconnectivity between two independent cultured neuronal networks. These results suggest that our method is applicable to network and plasticity analysis of neuronal networks.     

铭琪加特纳维尔癌症关顾中心

正铭琪癌症关顾中心旨在为癌症患者以及他们的家人和朋友提供实际上和情绪上的支援服务。自1996年第一所铭琪中心在爱丁堡成立以来,铭琪癌症关顾中心基金会的规模日益扩展,并透过委托世界知名的建筑师发展一系列开创先河的建筑。不论是刻意还是偶然的,当代建筑有时难免会产生冷漠或拒人于千里之外的观感,但每一所铭琪中心——在格拉斯哥、伦敦抑或香港——都希望营造令人宾至如归的感觉,