Use of hydrogen/deuterium exchange mass spectrometry and mutagenesis as a tool to identify the binding region of inhibitors targeting the human mitotic kinesin Eg5

An experimental procedure associating both hydrogen/deuterium exchange mass spectrometry (H/D-MS) and mutagenesis was developed to identify the protein-binding region of small inhibitors targeting the motor domain of the human mitotic kinesin Eg5. All the tested inhibitors decrease the deuterium incorporation rate of the same peptides corresponding to the following secondary structure elements: loop L5/helix alpha2 (region Tyr125-Glu145) and strand beta5/helix alpha3 (region Ile202-Leu227). Replacement of these two regions by the equivalent ones from N. crassa conventional kinesin heavy chain completely abolishes the modification of the deuterium incorporation rate by the inhibitors as well as their effects on the basal ATPase activity. The six tested inhibitors thus share a common binding site on Eg5. The strategy reported here allows the regions of a protein involved in ligand binding to be rapidly pinpointed and can be applied to other proteins and used as a general in vitro screening procedure to identify compounds targeting specific binding regions.     

河南省连康山自然保护区木本植物的研究

连康山自然保护区位于大别山北坡。这里地形复杂,水热资源丰富,次生森林植被发育良好,有木本植物78科,195属,437种及变种,是研究亚热带向暖温带过渡地带植物区系和森林生态系统的理想场所。根据野外调查、采集和鉴定、整理的结果,本文分析研究了本区的木本植物区系,木本植物资源,主要森林植被类型以及珍稀濒危树种,并提出了关于保护,引种栽培、发展和开发利用资源植物的建议,以增强保护区的活力,适应社会主义市场经济体制建设的需要。     

广西龙桥森林生态效益的研究

1982～1992年,对广西龙桥林区不同森林类型及空旷地的主要气象水文要素及凋落物进行了定位对比观测研究。揭示了该区不同森林类型调节气侯、涵养水源、保持水土及凋落物营养元素归还等生态效应。为该区的森林经营、效益评估及环境保护提供了依据和建议。     

抚顺市区及环城林带气挟菌分布的研究

本文研究了抚顺市区及环城林带不同环境位置大气微生物分布情况。实验结果显示：市区大气中细菌浓度明显高于环城林带地区；在环城林带地区，林带外大气中的细菌浓度也明显高于林带内。霉菌的分布规律基本与细菌相同。放线菌在市区与环城带地区分布的差别不大。环城林带地区空气清洁，而市区空气污染严重，尤其是繁忙的交通路口和繁华的商业区受气挟菌污染尤为严重。     

Denaturant sensitive regions in creatine kinase identified by hydrogen/deuterium exchange

The GdmHCl-induced unfolding of creatine kinase (CK) has been studied by hydrogen/deuterium (H/D) exchange combined with mass spectrometry. MM-CK unfolded for various periods in different denaturant concentrations was pulsed-labeled with deuterium to identify different conformational intermediate states. For all denaturation times or GdmHCl concentrations, we observed variable proportions of only two species. The low-mass envelope of isotope peaks corresponds to a species that has gained about 10 deuteriums more than native CK, and the high-mass envelope to a completely deuterated species. To localize precisely the unfolded regions in the states highly populated during denaturation, the protein was digested with two proteases (pepsin and type XIII protease) after H/D exchange and rapid quenching of the reaction. The two sets of fragments obtained were analyzed by liquid chromatography coupled to mass spectrometry to determine the deuterium level in each fragment. Bimodal distributions of deuterium were found for most peptides, indicating that these regions were either folded or unfolded. This behavior is consistent with cooperative, localized unfolding. However, we observed a monomodal distribution of deuterium in two regions (1-12 and 162-186). We conclude that the increment of mass observed in the low-mass species of the intact protein (+10 Da) has its origin in these two segments. These regions, which are very sensitive to low GdmHCl concentrations, are involved in the monomer-monomer interface of CK and their perturbation is likely to weaken the dimeric structure. At higher denaturant concentration, this would induce dissociation of the dimer.     

Cosserat continuum modelling of grain size effects in metal polycrystals

The global response of polycrystalline aggregates is investigated, in order to simulate grain size effects in IF ferritic steels. The mechanics of generalized continua is used to describe the studied phenomena. The polycrystal is regarded as a heterogeneous Cosserat medium, and the overall properties are estimated using a specific homogenization technique. To illustrate the capabilities of the model, some simple bidimensionnal computations are presented for different grain sizes. Afterwards tridimensionnal computations are shown in order to extract the global effect on the mechanical behaviour for grain sizes ranging from 5 µm to 120 µm. The finite element response is harder for the smallest grain size, but the model still underestimate the grain size effect on the tensile response. (© 2005 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)     

Strongly nonlinear modal equations for nearly integrable PDEs

Summary The purpose of this paper is the derivation of reduced, finite-dimensional dynamical systems that govern the near-integrable modulations ofN-phase, spatially periodic, integrable wavetrains. The small parameter in this perturbation theory is the size of the nonintegrable perturbation in the equation, rather than the amplitude of the solution, which is arbitrary. Therefore, these reduced equations locally approximate strongly nonlinear behavior of the nearly integrable PDE. The derivation we present relies heavily on the integrability of the underlying PDE and applies, in general, to anyN-phase periodic wavetrain. For specific applications, however, a numerical pretest is applied to fix the truncation orderN. We present one example of the reduction philosophy with the damped, driven sine-Gordon system and summarize our present progress toward application of the modulation equations to this numerical study.     

Influence of femtosecond laser marking on the corrosion resistance of stainless steels

Marking is of prime importance in the field of biomaterials to allow the identification of surgical tools as well as prostheses. Nowadays, marking is often achieved by means of laser beam, which may modify the characteristics of the treated surfaces. The use of laser devices delivering nanosecond pulses is known to induce dramatic corrosion degradations during sterilization or decontamination processes of the biomaterials. The aim of the present study is to investigate the ability of femtosecond (pulse duration in the 10⁻¹⁵ s range) laser treatments to avoid preferential corrosion processes of the marked areas, in order to extend the durability and the reliability of biomaterials. Experiments have been performed on martensitic Z30C13 and austenitic 316L stainless steels. Electrochemical measurements (cyclic polarization curves) were carried out to determine the passive state of samples before and after engraving, their corrosion rate and their susceptibility to localized corrosion. Further protracted immersion tests were also carried out to evaluate the natural long-term degradation of engraved parts. The electrochemical behavior is then explained on the basis of surface characterizations. Femtosecond laser marking is shown to provide an electrochemical ennoblement. Moreover, the chemical composition is not affected so that the passive character of both stainless steels is maintained, even improved if we consider the susceptibility to localized corrosion.     

Laser spectroscopy of cooled zirconium fission fragments

The first on-line laser spectroscopy of cooled fission fragments is reported. The r ions, produced in uranium fission, were extracted and separated using an ion guide isotope separator. The ions were cooled and bunched for collinear laser spectroscopy by a gas-filled linear Paul trap. New results for nuclear mean-square charge radii, dipole, and quadrupole moments are reported across the N=60 shape change. The mean-square charge radii are found to be almost identical to those of the Sr isotones and previously offered modeling of the radial changes is critically reviewed.     

Direct and inverse modeling for stochastic passive microbead rheology

The need in biology to understand the rheological properties of cells, tissues, membranes, and biological liquids, has led to new techniques in microrheology, where passive bead tracking is the fundamental probe of viscoelastic properties. We present stochastic methods for direct and inverse characterization based on noisy time series associated with either one or multiple beads, focusing on the distinctions in bead statistics due to bead-bead hydrodynamic interactions versus the ratio of bead separation distance and bead radii. Our modeling combines the seminal works of Mason & Weitz for single bead statistics and of Crocker-Levine-Lubensky for 2-bead statistics, together with the recent methods of the authors on direct and inverse simulation tools for generalized Langevin equations. The aim is to develop statistically accurate and fast direct simulation tools for bead-bead fluctuations, and then inversion methods derived from maximum likelihood methods and the Kalman filter. The application is to support experiments by R. Superfine and D. Hill and for characterization and simulations of transport mechanisms in lung airway surface liquids within the Virtual Lung Project at UNC Chapel Hill. (© 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)