Cell-Free Synthesis of Selenoproteins in High Yield and Purity for Selective Protein Tagging

The selenol group of selenocysteine is much more nucleophilic than the thiol group of cysteine. Selenocysteine residues in proteins thus offer reactive points for rapid post-translational modification. Herein, we show that selenoproteins can be expressed in high yield and purity by cell-free protein synthesis by global substitution of cysteine by selenocysteine. Complete alkylation of solvent-exposed selenocysteine residues was achieved in 10 minutes with 4-chloromethylene dipicolinic acid (4Cl-MDPA) under conditions that left cysteine residues unchanged even after overnight incubation. Gd^III−Gd^III distances measured by double electron–electron resonance (DEER) experiments of maltose binding protein (MBP) containing two selenocysteine residues tagged with 4Cl-MDPA-Gd^III were indistinguishable from Gd^III−Gd^III distances measured of MBP containing cysteine reacted with 4Br-MDPA tags.     

Oxidation of oleuropein studied by EPR and spectrophotometry

The autoxidation at alkaline pH and enzymatic oxidation by mushroom tyrosinase of oleuropein, the dominant biophenol present in the fruits and leaves of Olea europea, was followed by both electron paramagnetic resonance (EPR) and absorption spectroscopy. For comparison, the same oxidation processes were applied to 4‐methylcatechol, a simple polyphenol present in olive mill wastewaters. EPR spectra of stable o‐semiquinone radicals produced during autoxidation at pH 12 and short‐lived o‐semiquinone free radicals produced during autoxidation at pH 9.0 or tyrosinase action and stabilized by chelation with a diamagnetic metal ion (Mg²⁺) were recorded for both polyphenols, and the corresponding hyperfine splitting constants were determined. The UV‐Vis spectral characteristics of the oxidation of polyphenols were highly dependent on the type of polyphenol, oxidant type and the pH of the reaction. The kinetic behavior of tyrosinase in the presence of oleuropein and 4‐methylcatechol was followed by recording spectral changes at 400 nm (absorption maximum) over time. The tysosinase activity with oleuropein showed a pronounced pH optimum at pH 6.5 and a minor one around pH 8. From the data analysis of the initial rate at pH 6.5, the kinetic parameters K_m = 0.34 ± 0.03 mM and V_max = 0.029 ± 0.002 ΔA₄₀₀ min^–1 were determined for oleuropein.     

PARAMAGNETIC MEISSNER EFFECTS IN MELT-TEXTURED Y_(0.99)Nd_(0.01)Ba_2Cu_3O_(7-y)SUPERCONDUCTOR

The paramagnetic Meissner effect (PME) was observed on the field-cooled M-T curvesof melt-textured Y0.99Nd0.01Ba2Cu3O7-y in applied magnetic fields from 1kOe toJ0kOe. In the magnetic field below 1kOe, the sample undergoes the typical transi-tion from the normal state to the superconducting state when it was cooled down. Theapplied magnetic fields in which the PME occurs, are very high compared with thefields reported in literatures. We propose that the occurrence of the PME may be dueto a phase transition of flux vortices in the superconductors.     

掺锰BaTiO3 PTCR陶瓷的电子顺磁共振研究

利用电子顺磁共振（EPR）方法对Mn掺杂的BaTiO3 PTCR陶瓷在120－450K的温度范围内进行了研究，研究结果表明：高自旋Mn^2 离子的6重谱线非常明显，表征谱线位置的g因子为2．009 9,Mn^2 离子信号弱弱与BaTiO3晶体结构密切相关，Mn^2 离子信号的温度分布呈U型，即在三方相和立方相时，信号强度较强，而在四方相和斜方相时信号很弱，这是因为在四方相和斜方相中，由于BaTiO3晶体结构的扭曲和畸变，导致Mn^2 和Vo空位缔合生成Mn^2 -Vo复合缺陷，未成对电子的消失以及复合缺陷n^2 -Vo的生成最Mn^2 在四方相和斜方相（200－380K）消失的主因，实验中没有发现Mn^3 和Mn^4 的信号，而与实验的环境条件一致，掺Mn后样品Curie点的漂移可能是Mn进入晶格的一个判据，此外，由于试样的n(Ba)/n(Ti)=1，因而没有发现钡空位VBa及其它的复合缺陷（如：VBa-Mn^2 ,Mn^3 -Vo,La^3 -Mn^3 等），Mn^2 离子等电子陷阱的生长是Curie温度处电阻骤然增加的主要原因。     

EPR study of hydrogen-related defects in boron-doped p-type CVD homoepitaxial diamond films

Boron-doped p-type single crystalline chemical vapor deposition (CVD) homoepitaxial diamond films were investigated by electron paramagnetic resonance (EPR). Carbon dangling bond defects, which were accompanied by a nearby hydrogen atom, were observed in boron-doped p-type CVD diamond films on a IIa substrate similar to those observed in undoped diamond. This result suggested that the energy level position of the defects is located below the Fermi energy of boron-doped diamond, at around 0.3 eV above the valence-band top. The reason why the Fermi energy could be changed by the incorporation of boron atoms at low density (10¹⁶–10¹⁷/cm³) in the film in spite of the existence of the large defect density of EPR centers (10¹⁸/cm³) is thought to be that the singly occupied electron states of defects are located near the band edge. As for the thermal annealing effect of the defects, it was revealed that the concentration of the defects and the mobility of the p-type film did not change after annealing up to 1200 °C which is much higher than the temperature of boron–hydrogen pair dissociation.     

Identification of copper(II) and copper(I) and their interconversion in Cu/ZSM-5 De-NOx catalysts

A combined Fourier transform IR (FT-IR) and electron paramagnetic resonance (EPR) study shows that copper in ‘excessively exchanged’ Cu/ZSM-5 is initially present as OH bridged Cu²⁺ dimers, besides isolated Cu²⁺ ions. Upon heating, the dimers lose water and become oxygen bridged [Cu---O---Cu]²⁺ complexes. These are ‘EPR-silent’, presumably as a consequence of antiferromagnetic coupling of the unpaired electrons in each Cu²⁺; they are, however, detectable by their perturbation of the lattice vibrations, detected by a FT-IR band at 918–923 cm⁻¹. Reduction by hydrogen or carbon monoxide converts the [Cu---O---Cu]²⁺ complexes to pairs of Cu⁺ ions, while the color changes from green to grey. Reductive adsorption of nitrogen monoxide on Cu²⁺ results in the formation of Cu⁺---NO⁺. Destructive thermal desorption of nitrogen monoxide at 100°C not only restores the Cu²⁺ ions, but also appears to regenerate the [Cu---O---Cu]²⁺ complex. The results suggest that pairs of copper ions are instrumental in the catalytic decomposition of nitrogen monoxide.     

Photoinduced phenomena in nanostructured porous silicon

In this work we study the evolution of porous silicon photoluminescence under illumination. Samples were obtained by electrochemical etching of crystalline silicon wafers of different types. For the p-type samples the evolution of the spectra is explained in terms of photoinduced oxidation of nanostructures, which in turns leads to a discrete change in the photoluminescence spectra, as we reported in previous works. For the n-type material, a progressive decrease of the luminescence intensity is observed, which is attributed to the photoinduced generation of dangling bond related defect states at the surface layer surrounding the nanostructures. This model explains qualitatively well the kinetics of the evolution of the measured photoluminescence. Preliminary results of electronic paramagnetic resonance spectroscopy agree with this model.     

超导磁体系统产生的磁场作用下的微重力环境

超导磁体系统产生的磁场力可抑制晶体生长过程中的自然对流,从而提高晶体结晶质量。通过理论分析,得到了实现微重力环境所要求的磁场强度分布。针对特定结构的超导磁体系统,数值模拟得到了不同晶体结晶过程中自然对流完全抑止所需要的线圈电流密度,以及工作空间内的磁场强度和磁加速度。以圆筒内水的自然对流为例,分别得到了重力作用下的流场和温度场,以及超导磁体系统微重力区内的流场和温度场。结果表明:通过特定的超导磁体结构和不同电流密度,可实现不同晶体结晶过程的微重力环境,达到抑止自然对流的目的。     

顺磁氧分析仪在转炉煤气回收系统的应用

顺磁氧分析仪在转炉煤气回收系统中的应用是种新尝试，简要介绍了顺磁氧分析仪的测量原理、基本构成、基本特点及系统控制原理，从而确定顺磁氧分析仪在转炉煤气回收系统中的可行性。     

Role of Nitric Oxide-Derived Metabolites in Reactions of Methylglyoxal with Lysine and Lysine-Rich Protein Leghemoglobin

Carbonyl stress occurs when reactive carbonyl compounds (RCC), such as reducing sugars, dicarbonyls etc., accumulate in the organism. The interaction of RCC carbonyl groups with amino groups of molecules is called the Maillard reaction. One of the most active RCCs is α-dicarbonyl methylglyoxal (MG) that modifies biomolecules forming non-enzymatic glycation products. Organic free radicals are formed in the reaction between MG and lysine or Nα-acetyllysine. S-nitrosothiols and nitric oxide (^•NO) donor PAPA NONOate increased the yield of organic free radical intermediates, while other ^•NO-derived metabolites, namely, nitroxyl anion and dinitrosyl iron complexes (DNICs) decreased it. At the late stages of the Maillard reaction, S-nitrosoglutathione (GSNO) also inhibited the formation of glycation end products (AGEs). The formation of a new type of DNICs, bound with Maillard reaction products, was found. The results obtained were used to explain the glycation features of legume hemoglobin—leghemoglobin (Lb), which is a lysine-rich protein. In Lb, lysine residues can form fluorescent cross-linked AGEs, and ^•NO-derived metabolites slow down their formation. The knowledge of these processes can be used to increase the stability of Lb. It can help in better understanding the impact of stress factors on legume plants and contribute to the production of recombinant Lb for biotechnology.