Rapid NMR Assignments of Proteins by Using Optimized Combinatorial Selective Unlabeling

A new approach for rapid resonance assignments in proteins based on amino acid selective unlabeling is presented. The method involves choosing a set of multiple amino acid types for selective unlabeling and identifying specific tripeptides surrounding the labeled residues from specific 2D NMR spectra in a combinatorial manner. The methodology directly yields sequence specific assignments, without requiring a contiguously stretch of amino acid residues to be linked, and is applicable to deuterated proteins. We show that a 2D N-15,H-1]HSQC spectrum with two 2D spectra can result in approximate to 50% assignments. The methodology was applied to two proteins: an intrinsically disordered protein (12kDa) and the 29kDa (268 residue) -subunit of Escherichia coli tryptophan synthase, which presents a challenging case with spectral overlaps and missing peaks. The method can augment existing approaches and will be useful for applications such as identifying active-site residues involved in ligand binding, phosphorylation, or protein-protein interactions, even prior to complete resonance assignments.

Stimuli-responsive colorimetric and NIR fluorescence combination probe for selective reporting of cellular hydrogen peroxide

Hydrogen peroxide (H2O2) is a key reactive oxygen species and a messenger in cellular signal transduction apart from playing a vital role in many biological processes in living organisms. In this article, we present phenyl boronic acid-functionalized quinone-cyanine (QCy-BA) in combination with AT-rich DNA (exogenous or endogenous cellular DNA), i.e., QCy-BA subset of DNA as a stimuli-responsive NIR fluorescence probe for measuring in vitro levels of H2O2. In response to cellular H2O2 stimulus, QCy-BA converts into QCy-DT, a one-donor-two-acceptor (D2A) system that exhibits switch-on NIR fluorescence upon binding to the DNA minor groove. Fluorescence studies on the combination probe QCy-BA subset of DNA showed strong NIR fluorescence selectively in the presence of H2O2. Furthermore, glucose oxidase (GOx) assay confirmed the high efficiency of the combination probe QCy-BA subset of DNA for probing H2O2 generated in situ through GOx-mediated glucose oxidation. Quantitative analysis through fluorescence plate reader, flow cytometry and live imaging approaches showed that QCy-BA is a promising probe to detect the normal as well as elevated levels of H2O2 produced by EGF/Nox pathways and post-genotoxic stress in both primary and senescent cells. Overall, QCy-BA, in combination with exogenous or cellular DNA, is a versatile probe to quantify and image H2O2 in normal and disease-associated cells.

On the issues of resolving a low melting combination as a definite eutectic or an elusive cocrystal: A critical evaluation

Cocrystals and eutectics are different yet related crystalline multi-component adducts with diverse applications in pharmaceutical and materials fields. Recently, they were shown to be alternate products of cocrystallization experiments. Whereas a cocrystal shows distinct diffraction, spectroscopic and thermal signatures as compared to parent components, the hallmark of a eutectic is its low melting nature. However, in certain cases, there can be a problem when one resorts to design a cocrystal and assess its formation vis-A -vis a eutectic. In the absence of a gold standard method to make a cocrystal, it is often difficult to judge how exhaustive should the cocrystallization trials be to ensure the accomplishment of a desired/putative cocrystal. Further, a cocrystal can manifest with intermolecular interactions and/or crystal structure similar to that of its parent compounds such that the conventional diffraction and spectroscopic techniques will be of little help to conclusively infer the formation of cocrystal in the lack of single crystals. Such situations combined with low melting behavior of a combination brings the complication of resolving the combination as a cocrystal or eutectic since now both the adducts share common features. Based on the curious case of Caffeine-Benzoic acid combination, this study aims to unfold the intricate issues related to the design, formation and characterization of cocrystals and eutectics for a way forward. The utility of heteronuclear seeding methodology in establishing a given combination as a cocrystal-forming one or a eutectic-forming one in four known systems is appraised.

Numerical Analysis of a Dual Polarization Mode-Locked Laser with a Quarter Wave Plate

Dynamical behaviors and frequency characteristics of an active mode-locked laser with a quarter wave plate (QWP) are numerically studied by using a set pf vectorial laser equation. Like a polarization self-modulated laser, a frequency shift of half the cavity mode spacing exists between the eigen-modes in the two neutral axes of QWP. Within the active medium, the symmetric gain and cavity structure maintain the pulse's circular polarization with left-hand and right-hand in turn for each round trip. Once the left-hand or right-hand circularly polarized pulse passes through QWP, its polarization is linear and the polarized direction is in one of the directions of i45o with respect to the neutral axes of QWP. The output components in the directions of i45" from the mirror close to QWP are all linearly polarized with a period of twice the round-trip time.

Polarization Self-Modulation Phenomenon in a Free Oscillated Nd: YAG Laser

Polarization self-modulation effect in a free oscillated Nd:YAG laser is investigated after a quarter wave plate is introduced independently in the two positions of the cavity. As described in the previous experiments, the intensity components in the orthogonal directions are modulated with a period of the round-trip time or twice. Different pulse shapes reveal that the seed field from the spontaneous emission is not uniform and seems to be stochastic for each pulse.

Surface amorphous and crystalline microstructure by alloying zirconium using Nd : YAG pulsed laser

A novel composite coating was synthesized by laser alloying of zirconium nanoparticles on an austenite stainless steel surface using a pulsed Nd:YAG laser. The coating contained duplex microstructures comprising an amorphous phase and an austenitic matrix. A discontinuous zirconium-containing region formed at a depth of 16 mum below the surface. The amorphous phase was present in the zirconium-rich region, with the composition of zirconium ranging from 7.8 to 14.5 at. pet. The formation of the amorphous phase was attributed to the zirconium addition. The hardness, corrosion, and wear-corrosion resistance of the irradiated coating were evidently enhanced compared to those of the stainless steel.

Influence of Local Magnetic Fields on P-Doped Si Floating Zone Melting Crystal Growth in Microgravity

A two-dimensional axisymmetric numerical model is presented to study the influence of local magnetic fields on P-doped Si floating zone melting crystal growth in microgravity. The model is developed based on the finite difference method in a boundary-fitted curvilinear coordinate system. Extensive numerical simulations are carried out, and parameters studied include the curved growth interface shape and the magnetic field configurations. Computed results show that the local magnetic field is more effective in reducing the impurity concentration nonuniformity at the growth interface in comparison with the longitudinal magnetic field. Moreover, the curved growth interface causes more serious impurity concentration nonuniformity at the growth interface than the case with a planar growth interface.

Microstructure and wear resistance of in situ TiCp composite coating by laser cladding

A new in situ method was realized by one step laser cladding to produce Ni-base alloy composite coating reinforced by in situ reacted and gradiently distributed TiCp particles. The submicron TiCp particles were formed and uniformly distributed because of the in situ reaction and trapping effect under the rapid solidification condition. And, TiCp particles were of gradient distribution on a macro scale and their volume fraction increased from 1.86% at the layer/substrate interface to a maximum 38.4% at the surface of the layer. Furthermore, the in situ generated TiCp/gamma-Ni interfaces were free from deleterious surface reactions. Additionally, the clad coating also revealed a high microhardness of gradient variation with the layer depth and the superior abrasive wear resistance.

Intensity Distribution Design for Laser-Induced Thermal Loading Based on Numerical Simulation

To accomplish laser-induced thermal loading simulation tests for pistons,the Gaussian beam was modulated into multi-circular beam with specific intensity distribution.A reverse method was proposed to design the intensity distribution for the laser-induced thermal loading based on finite element(FE) analysis.Firstly,the FE model is improved by alternating parameters of boundary conditions and thermal-physical properties of piston material in a reasonable range,therefore it can simulate the experimental resul...

Effects of La2O3 on Microstructure and Wear Properties of Laser Clad gamma/Cr7C3/TiC Composite Coatings on TiAl Intermatallic Alloy

The effects of La2O3 addition on the microstructure and wear properties of laser clad gamma/C(r)7C(3)/TiC composite coatings on gamma-TiAl intermetallic alloy substrates with NiCr-Cr3C2 precursor mixed powders have been investigated by optical microscopy (OM), scanning electron microscopy (SEM), X-ray diffraction (XRD) and energy-dispersive spectrometer (EDS) and block-on-ring wear tests. The responding wear mechanisms are discussed in detail. The results are compared with that for composite coating without La2O3. The comparison indicates that no evident new crystallographic phases are formed except a rapidly solidified microstructure consisting of the primary hard Cr7C3 and TiC carbides and the gamma/Cr7C3 eutectics distributed in the tough gamma nickel solid solution matrix. Good finishing coatings can be achieved under a proper amount of La2O3-addition and a suitable laser processing parameters. The additions of rare-earth oxide La,03 can refine and purify the microstructure of coatings, relatively decrease the volume fraction of primary blocky Cr7C3 to Cr7C3/gamma eutectics, reduce the dilution of clad material from base alloy and increase the microhardness of the coatings. When the addition of La2O3 is approximately 4 wt.%, the laser clad composite coating possesses the highest hardness and toughness. The composite coating with 4 wt.%La2O3 addition can result the best enhancement of wear resistance of about 30%. However, too less or excessive addition amount of La2O3 have no better influence on wear resistance of the composite coating.

Numerical simulation of an adaptive optics system with laser propagation in the atmosphere

A comprehensive model of laser propagation in the atmosphere with a complete adaptive optics (AO) system for phase compensation is presented, and a corresponding computer program is compiled. A direct wave-front gradient control method is used to reconstruct the wave-front phase. With the long-exposure Strehl ratio as the evaluation parameter, a numerical simulation of an AO system in a stationary state with the atmospheric propagation of a laser beam was conducted. It was found that for certain conditions the phase screen that describes turbulence in the atmosphere might not be isotropic. Numerical experiments show that the computational results in imaging of lenses by means of the fast Fourier transform (FFT) method agree well with those computed by means of an integration method. However, the computer time required for the FFT method is 1 order of magnitude less than that of the integration method. Phase tailoring of the calculated phase is presented as a means to solve the problem that variance of the calculated residual phase does not correspond to the correction effectiveness of an AO system. It is found for the first time to our knowledge that for a constant delay time of an AO system, when the lateral wind speed exceeds a threshold, the compensation effectiveness of an AO system is better than that of complete phase conjugation. This finding indicates that the better compensation capability of an AO system does not mean better correction effectiveness. (C) 2000 Optical Society of America.

Microstructure of Zr-alloyed coating using pulsed laser

Coatings were synthesized by laser alloying of zirconium (Zr) particles using a pulsed Nd:YAG laser on an austenite stainless steel. The distribution of Zr is uneven, in a depth of 18 mum just below the surface of the pool. The coating consisted of duplex microstructures, i.e. metallic glass (MG) and austenite. The MG formed in Zr-contained regions, with the Zr composition ranging from 7.6 to 16.8 at.%. The formation of the MG was attributed to an increase in glass-forming ability by Zr addition.

Application of the Conjugate Gradient Optimization Algorithm of Neural Network on Laser Machining

在应用激光技术加工复杂曲面时,通常以采样点集为插值点来建立曲面函数,然后实现曲面上任意坐标点的精确定位。人工神经网络的BP算法能实现函数插值,但计算精度偏低,往往达不到插值精确要求,造成较大的加工误差。提出人工神经网络的共轭梯度最优化插值新算法,并通过实例仿真,证明了这种曲面精确定位方法的可行性,从而为激光加工的三维精确定位提供了一种良好解决方案。这种方法已经应用在实际中。