Proteome analysis of tissues by mass spectrometry

Tissues and biofluids are important sources of information used for the detection of diseases and decisions on patient therapies. There are several accepted methods for preservation of tissues, among which the most popular are fresh‐frozen and formalin‐fixed paraffin embedded methods. Depending on the preservation method and the amount of sample available, various specific protocols are available for tissue processing for subsequent proteomic analysis. Protocols are tailored to answer various biological questions, and as such vary in lysis and digestion conditions, as well as duration. The existence of diverse tissue‐sample protocols has led to confusion in how to choose the best protocol for a given tissue and made it difficult to compare results across sample types. Here, we summarize procedures used for tissue processing for subsequent bottom‐up proteomic analysis. Furthermore, we compare protocols for their variations in the composition of lysis buffers, digestion procedures, and purification steps. For example, reports have shown that lysis buffer composition plays an important role in the profile of extracted proteins: the most common are tris(hydroxymethyl)aminomethane, radioimmunoprecipitation assay, and ammonium bicarbonate buffers. Although, trypsin is the most commonly used enzyme for proteolysis, in some protocols it is supplemented with Lys‐C and/or chymotrypsin, which will often lead to an increase in proteome coverage. Data show that the selection of the lysis procedure might need to be tissue‐specific to produce distinct protocols for individual tissue types. Finally, selection of the procedures is also influenced by the amount of sample available, which range from biopsies or the size of a few dozen of mm² obtained with laser capture microdissection to much larger amounts that weight several milligrams.     

Divergent Synthesis of Novel Cylindrocyclophanes that Inhibit Methicillin-Resistant Staphylococcus aureus (MRSA)

The cylindrocyclophanes are a family of macrocyclic natural products reported to exhibit antibacterial activity. Little is known about the structural basis of this activity due to the challenges associated with their synthesis or isolation. We hypothesised that structural modification of the cylindrocyclophane scaffold could streamline their synthesis without significant loss of activity. Herein, we report a divergent synthesis of the cylindrocyclophane core enabling access to symmetrical macrocycles by means of a catalytic, domino cross-metathesis-ring-closing metathesis cascade, followed by late-stage diversification. Phenotypic screening identified several novel inhibitors of methicillin-resistant Staphylococcus aureus. The most potent inhibitor has a unique tetrabrominated [7,7]paracyclophane core with no known counterpart in nature. Together these illustrate the potential of divergent synthesis using catalysis and unbiased screening methods in modern antibacterial discovery.     

Isolation and identification of soil bacteria resistant to surfactants in washing detergents

Linear alkylbenzene sulfonate (LAS) and polyoxyethylene lauryl ether (POLE) are the major surfactants in washing detergents. In the present study, we isolated surfactant-resistant bacteria from soil samples collected from a sports ground and a farm field. The samples were treated with 2.0% LAS or POLE at 25°C for 30 min and cultivated on agar plates at 25°C for several days, after which manifold bacterial colonies were isolated. Thereafter, we tested the ability of each bacterial isolate to resist the antibacterial activity of the surfactant. Ten LAS-resistant strains were isolated, and all were found to be Gram-negative bacteria such as Enterobacter and Pseudomonas. On the other hand, 18 POLE-resistant strains were isolated, of which 14 were Gram-positive bacteria including Bacillus and Microbacterium. Notably, one POLE-resistant strain was identified as Bacillus cereus, a potential causative agent for foodborne illness. The genera of LAS- and POLE-resistant bacteria did not overlap. Therefore, the combination of LAS and POLE could be more effective to eliminate soil bacteria from clothes and/or daily necessities.     

适用于小型岩石试件的三轴压缩试验技术与设备的开发及其应用

为了用有限的岩石试样在室内进行大量的岩石力学试验测试，对直径10mm、长度20mm的小型岩石试件进行开发、制作。同时对S．Okubo等（2002）开发的透明围压容器进行改进，其围压可达10MPa左右。用此改进的试验设备对小型岩石试件进行三轴压缩试验。通过对田下凝灰岩的试验，由小型试件得到的力学参数——黏聚力和内摩擦角，与用常规试件获得的试验结果相一致。而且在载荷速率交替变换的试验中，用一个小型试件就可获得应力一应变曲线的载荷速率相关性。通过对三峡库区周边滑坡地带采集的岩石试样的测试，成功获得不同岩石的强度特性曲线及表示载荷速率相关性的参数。     

Aerosol Particle Collection Efficiency of Holey Carbon Film-Coated TEM Grids

A simple sampling method to collect aerosol particles for transmission electron microscope (TEM) analysis was developed by R’mili and others in 2013. The method involves passing air through a holey carbon film-coated copper mesh TEM grid (holey carbon grid) and sampling particles by filtration. In this study, we proposed a modified calculation method to represent the collection efficiencies of holey carbon grids, taking into consideration the porosity of the copper mesh. We then evaluated the particle collection efficiencies of holey carbon grids both theoretically and experimentally. We tested the collection efficiency of two types of holey carbon grids, with nominal pore sizes of 1.2 and 0.6 μm, using particles of monodispersed polystyrene latex (PSL) and potassium chloride. The overall collection efficiency of each grid (E_grid) was determined by the downstream/upstream concentration ratio measured by condensation particle counters (CPCs). In addition, for PSL particles, the collection efficiency of the holey carbon film (E_film) was determined by the ratio of the number of particles on the film (counted on a scanning electron microscope) to the number of inflow particles (counted by a CPC). We compared model calculations against the experimental results obtained in this study and those reported by R’mili and others in 2013. These data showed that the calculated E_grid values were in reasonably good agreement with the experimental E_grid values. However, although the model calculation indicated that E_film ≈ E_grid, there was an inconsistency between the experimental E_film and E_grid, which requires further investigation in order to determine its cause.

Copyright 2014 American Association for Aerosol Research     

Study on FBR core concepts to increase proliferation resistance of plutonium in LWR–FBR transition period

The possibility to enhance proliferation resistance of discharged plutonium in fast breeder reactor (FBR) has been investigated in terms of reactor core-design aspects. The provisional target for proliferation resistance measures based on Saito's attractiveness (ATTR) is defined. It is found that a few percent of plutonium loading and/or Am/Cm loading, which come from various types of spent fuel, might satisfy the provisional target with a minimum impact on the core neutronic performances. On Am/Cm loading core, decay heat constraints for fuel-handling aspects are found to be important and should be considered in design. There is not significant change on the current developing scenarios for light water reactor–FBR transition period by applying the measures based on Saito's ATTR. It is found that applying Kimura's proposal, 15% ²³⁸Pu content requires about 7% MA loading fraction both in the core and the blanket, and it only can be applied at limited case. The period to consume minor actinides is shorter than to consume plutonium.     

The influence of processing methods on the dielectric properties of BaTi1-xGdxO3-x/2 - Based materials

In this study, the intermediate rare-earth oxide Gd₂O₃ (Gd) was substituted in different amounts (x = 0.2–2 mol%) for the formulation of BaTi_1-xGd_xO_3-x/2 (BTG_x) dielectric materials. The effect of B-site substitution was confirmed by the additional Raman active A_1g octahedral peak at ~835cm^-1 strengthened at x ≥ 0.4 mol%. Additionally, properties of 0.9BTG_0.007-0.1BA dielectric ceramics were analysed based on the influence of various processing methods as a function of sintering temperature. The focal samples were labelled Method-A (direct-mix) and Method-B (indirect-mix). As the sintering temperature (1075–1200 °C) increased, the 1 kHz response of the ε–T curves of Method-A samples transformed from a single peak to broad-narrow double peaks of high dielectric loss tangent (tan δ). Nonetheless, samples of Method-B possessed a clearly defined transmission electron microscopy (TEM) core-shell structure, flattened double-peak ε-T curves, optimised dielectric properties (ε = ~1563–1851 and tan δ < 1.5% at room temperature), and a wide-ranging temperature behaviour that meets the X8R dielectric standards (ΔC/C_25°C < ±15%). The maximum dielectric breakdown strength of Method-B samples reached ~131 kVcm, while the energy storage density was ~0.726 J/cm³ at a maximum efficiency of ~80% at 1100 °C. Thus, exhibiting good potentials for balancing temperature stability with energy storage applications.     

Development of Motor Model of Rotor Slot Harmonics for Speed Sensorless Control of Induction Motor

This paper proposes a novel mathematical dynamic model to represent the steady‐state and transient‐state characteristics of rotor slot harmonics of an induction motor for sensorless control. Although it is well known that the rotor slot harmonics originate from the mechanical structure of the induction motor, a mathematical model that describes the relationship between the stator/rotor currents of the induction motor and the slot harmonics has not yet been proposed. Therefore, in this paper, a three‐phase model of the induction motor that depicts the rotor slot harmonics is developed by taking into consideration the magnetomotive force harmonics and the change in the magnetic air gap caused by the rotor slots. The validity of the proposed model is verified by comparing the experimental results and the calculated values. © 2013 Wiley Periodicals, Inc. Electr Eng Jpn, 186(3): 63–74, 2014; Published online in Wiley Online Library ( wileyonlinelibrary.com ). DOI 10.1002/eej.22478     

Design of Nonlinear Servo Systems by the Fuzzy Method

This paper presents the design of nonlinear servo systems using the fuzzy method. The idea behind this control method is to divide the operating region of a nonlinear system into a collection of local linear servo control systems and apply the fuzzy method for calculation of the control input. The control input of each local linear servo system is calculated using the Davison–Smith method and the Hikita pole assignment method. We can prove that all states of the fuzzy servo system are bounded and that the output errors converge to zero by the Lyapunov method. Simulations on a nonlinear mass‐spring‐damper system illustrate the effectiveness of the proposed method.