Development of Fluorogenic Probes for Rapid High-Contrast Imaging of Transient Nuclear Localization of Sirtuin 3

Protein labeling using fluorogenic probes enables the facile visualization of proteins of interest. Herein, we report new fluorogenic probes consisting of a rationally designed coumarin ligand for the live-cell fluorogenic labeling of the photoactive yellow protein (PYP)-tag. On the basis of the photochemical mechanisms of coumarin and the probe–tag interactions, we introduced a hydroxy group into an environment-sensitive coumarin ligand to modulate its spectroscopic properties and increase the labeling reaction rate. The resulting probe had a higher labeling reaction rate constant and a greater fluorescence OFF–ON ratio than any previously developed PYP-tag labeling probe. The probe enabled the fluorogenic labeling of intracellular proteins within minutes. Furthermore, we used our probe to investigate the localization of sirtuin 3 (SIRT3), a mitochondrial deacetylase. Although the nuclear localization of SIRT3 has been controversial, this transient nuclear localization was clearly captured by the rapid, high-contrast imaging enabled by our probe.     

Formation Mechanism and Synthesis of Apatite-Type Structure Ba2+xLa8−x(SiO4)6O2−δ

The formation process of Ba₂La₈(SiO₄)₆O₂ was clarified using thermogravimetry–differential thermal analysis (TG-DTA) and a high-temperature powder X-ray diffraction (HT-XRD) method. Phase changes identified from the HT-XRD data surprisingly corresponded to the weight loss and/or endothermic peaks observed in the TG-DTA curves. Raw material with the composition Ba₂La₈(SiO₄)₆O₂ was completely reacted at 1400°C and produced only an apatite-type compound without a secondary phase. Moreover, the synthesis of Ba_{2+ x} La_{8− x} (SiO₄)₆O_2−δ crystals with x = 0–2 was attempted using a solid-state reaction.     

Optimization of Set-up Conditions for Stability of The Centerless Grinding Process

The stability of the centerless grinding process is very sensitive to the set-up conditions due to the uniqueness of the work-holding system. Centerless grinding produces precision components with high productivity only when the set-up condition is optimally chosen. This paper describes the effect of set-up conditions on three stability criteria of the centerless grinding system. It also presents guidelines for determining proper set-up conditions to avoid spinners, chatter vibration and roundness problems. Finally, an algorithm for providing the optimum set-up condition based on process aims is proposed and the simulation results are discussed.     

深基坑工程动态施工反演分析与变形预报

提出了动态施工反演分析的思想，即在常规的反分析过程中引入逐步开挖和逐道支撑的动态施工因素，以求仿真模拟工程实际情况，进而为相继施工阶段的变形预报提供可靠保证。文中基于任意施工阶段间围护墙体变形与内支撑梁轴力的增量测值和施工ＦＥＭ优化反演分析法，反推了各土层的弹性模量系数，给出了作用在墙体两侧的土压力分布，并预报各相继施工阶段的墙体和土体变形、内力及内支撑梁轴力。     

What is the significance of nitrogen-vacancy centers in the doping into diamond?

In order to advance quantum computer after successful synthesis of a qubit (quantum-bit) using a NV (nitrogen-vacancy) center in diamond crystal, the innovation of a NOT processor is strongly demanded. A candidate of it can be one NV center associated with an additional N atom, which is called a NV–N center. Making use of a classical molecular dynamics simulation of N ion implantation into pure diamond with an energy of 200 eV, we have examined the crystallographic structure of “NV–N” centers. However, at low temperature of 30 K, most implanted N atoms became interstitials, thus it was difficult to identify even NV centers.     

Preparation of sharp-melting hard palm midfraction and its use as hard butter in chocolate

Preparation of hard palm midfractions (PMF) and its use as a cocoa butter equivalent ingredient were studied. Hard PMF is obtained by multistep fractionation of palm oil involving dry fractionation (DF) and/or solvent fractionation (SF), usually using hexane or acetone. From our experience, in acetone, a polar solvent, symmetrical 1,3-disaturated triacylglycerols tend to selectively crystallize more than nonsymmetrical 1,2- or 2,3-disaturated triacylglycerols, making it suitable for obtaining the solid midfraction. Unfortunately, triacylglycerols are very soluble in hexane, and temperatures at least 15 degrees lower than those required for acetone must be used for equivalent crystal yields. On the other hand, DF is a less expensive and safer process. Thus, multistep fractionation combining DF and SF using acetone was developed to achieve sufficient removal of high-melting components, and further enrichment of 1,3-dipalmitoyl-2-oleoylglycerol and the hard PMF was obtained by triple-step fractionation of palm olein or double-step fractionation of soft PMF. Compared to conventional hard PMF, this hard PMF had a steeper melting curve and better snapping and sharp-melting qualities when used in chocolate. Heat resistance of the hard PMF chocolate was similar to the conventional hard PMF chocolate, and its bloom resistance could be improved by adding polyglycerol fatty acid esters.     

The mechanism of formation of iron oxide and oxyhydroxides in aqueous solutions at room temperature

For a better understanding of the atmospheric rusting of iron and steels, the present work is aimed to explore the mechanism of formation of green rusts, Fe₃O₄, α-FeOOH, β-FeOOH, γ-FeOOH, δ-FeOOH and amorphous ferric oxyhydroxide in aqueous solution at room temperature. The formation processes on which end products are determined are strongly affected by the oxidation rate, pH and the structure and composition of initial and intermediate species of iron. The systematic diagram of formation processes of iron oxide and oxyhydroxides has been presented, in which both dissolved and solid species of iron are included.     

The mechanism of atmospheric rusting and the protective amorphous rust on low alloy steel

Atmospheric rusting of mild and low alloy steels was studied by means of infra-red and far infra-red spectrophotometries, X-ray and electron diffraction methods and scanning electron microscopy. The rusting process can be interpreted on the basis of a previously reported diagram for rust formation in aqueous solution.A large amount of amorphous matter in rust formed in semi-rural atmosphere was identified by infra-red and far infra-red spectra as amorphous ferric oxyhydroxide, FeO_x (OH)_8–2x. The amorphous ferric oxyhydroxide rust on low alloy steel was dense and uniform, and contained a considerable amount of bound water. From these results it can be concluded that the amorphous ferric oxyhydroxide rust acts as a protective barrier against atmospheric rusting of the steels. Cu, P and Cr in low-alloy steels are inferred to favour the formation of crack-free, uniform rust layer and help to produce uniform amorphous ferric oxyhydroxide.     

X-ray photoelectron spectrum of Fe2+ state in iron oxides

As a standard for identification of iron oxides by X-ray photoelectron spectroscopy, Fe²⁺ spectra are extracted from mixed Fe 2p_3/2 spectra of Fe³⁺, Fe³⁺ and metallic states. The peaks of Fe²⁺ spectra are all located at binding energy of 708·5 eV. The width of Fe²⁺ spectrum seems to be dependent on crystallinity, and is 2·2 eV for a bulk crystalline oxide and 2·9 eV for an amorphous thin film under instrumental condition with FWHM of 1·3 eV for Au 4f_7/2.