Cytosine-5 methylation-directed construction of a Au nanoparticle-based nanosensor for simultaneous detection of multiple DNA methyltransferases at the single-molecule level

DNA methylation at cytosine/guanine dinucleotide islands (CpGIs) is the most prominent epigenetic modification in prokaryotic and eukaryotic genomes. DNA methyltransferases (MTases) are responsible for genomic methylation, and their aberrant activities are closely associated with various diseases including cancers. However, the specific and sensitive detection of multiple DNA MTases has remained a great challenge due to the specificity of the methylase substrate and the rareness of methylation-sensitive restriction endonuclease species. Here, we demonstrate for the first time the cytosine-5 methylation-directed construction of a Au nanoparticle (AuNP)-based nanosensor for simultaneous detection of multiple DNA MTases at the single-molecule level. We used the methyl-directed endonuclease GlaI to cleave the site-specific 5-methylcytosine (5-mC). In the presence of CpG and GpC MTases (i.e., M.SssI and M.CviPI), their hairpin substrates are methylated at cytosine-5 to form the catalytic substrates for GlaI, respectively, followed by simultaneous cleavage by GlaI to yield two capture probes. These two capture probes can hybridize with the Cy5/Cy3–signal probes which are assembled on the AuNPs, respectively, to form the double-stranded DNAs (dsDNAs). Each dsDNA with a guanine ribonucleotide can act as the catalytic substrate for ribonuclease (RNase HII), inducing recycling cleavage of signal probes to liberate large numbers of Cy5 and Cy3 molecules from the AuNPs. The released Cy5 and Cy3 molecules can be simply quantified by total internal reflection fluorescence (TIRF)-based single-molecule imaging for simultaneous measurement of M.SssI and M.CviPI MTase activities. This method exhibits good specificity and high sensitivity with a detection limit of 2.01 × 10⁻³ U mL⁻¹ for M.SssI MTase and 3.39 × 10⁻³ U mL⁻¹ for M.CviPI MTase, and it can be further applied for discriminating different kinds of DNA MTases, screening potential inhibitors, and measuring DNA MTase activities in human serum and cell lysate samples, holding great potential in biomedical research, clinical diagnosis, drug discovery and cancer therapeutics.

Cytosine-5 methylation-directed construction of Au nanoparticle-based nanosensors enables specific and sensitive detection of multiple DNA methyltransferases.     

Diastereoselective Cyclobutenol Synthesis: A Heterogeneous Palladium-Catalyzed Oxidative Carbocyclization-Borylation of Enallenols

A highly selective and efficient oxidative carbocyclization/borylation of enallenols catalyzed by palladium immobilized on amino-functionalized siliceous mesocellular foam (Pd-AmP-MCF) was developed for diastereoselective cyclobutenol synthesis. The heterogeneous palladium catalyst can be recovered and recycled without any observed loss of activity or selectivity. The high diastereoselectivity of the reaction is proposed to originate from a directing effect of the enallenol hydroxyl group. Optically pure cyclobutenol synthesis was achieved by the heterogeneous strategy by using chiral enallenol obtained from kinetic resolution.     

One-step method for preparing the experimental pure state in nuclear magnetic resonance

Quantum computing is an emerging and promising research field in modern quantum physics and is committed to accelerating traditional calculation processes using the principle of quantum mechanics[1,2].Several computation models have been proposed,including the circuit model[3],topologic quantum computation[4],adiabatic quantum computing[5],and duality quantum computing[6-8].Universal quantum computation generally includes a system initialization,evolutionary control,and a final-state readout[9],where the preparation of the initial state is a key step in the entire process.     

Identification and quantification of main flavonoids in the leaves of Bambusa multiplex cv. Fernleaf

Abstract

The chemical profile of Bambusa multiplex cv. Fernleaf (B. multiplex) leaves was analysed by UPLC-DAD-Q-TOF-MS. Twelve compounds were identified and C-glycosyl flavonoids, including vitexin, isovitexin, isoorientin and its derivatives, are the main constitutes of the plant. Besides, a HPLC method for isoorientin quantification was developed. The RSD of retention time and peak area were 0.05% and 2.04% for six times analysis of isoorientin with concentration of 20?μg/mL. The recovery of isoorientin in real sample was 99.2%. The general trend of isoorientin content in B. multiplex leaves was that it steady increased from Jan. to May, and then quickly decreased. The maximum was found on May with value of 4.7?mg/g. The lowest level of isoorientin was found during Aug. to Nov. with value of about 1.66?mg/g. In different seasons, isoorientin is always the most dominant flavonoid which was accounted for about 50% of total flavonoids in the sample.     

基于非线性光纤环形镜的少模脉冲幅度调制再生器

随着网络带宽需求的快速增加,波分复用系统的容量已接近非线性香农极限.为了适应未来网络的发展,空分复用技术引起了越来越多的关注.本文首次提出基于少模非线性光纤环形镜(FM-NOLM)的脉冲幅度调制(PAM)全光再生器,描述了其工作原理和具体设计过程.采用COMSOL软件对组成FM-NOLM的硫化物高非线性光纤进行了模式特性仿真.以LP01,LP11,LP21三个光纤模式为例,确定了再生器的参数,计算出每个模式的功率转移函数曲线.仿真分析了该少模PAM-4全光再生器的噪声抑制(NRR)性能,并与单模情形进行了比较.研究表明,1)对于每个空间模式的PAM信号,所有再生电平具有一致的功率转移性能;2)当输入信噪比(SNR)约大于20 dB时,三种模式的噪声抑制比均可超过3 dB,并随着输入信噪比线性增加,其斜率约为1.2;3)在相同输入SNR条件下,三种模式的噪声抑制比相差不大,不超过1.1 dB.为了说明再生器的再生性能,当输入SNR为25 dB时,我们还给出了再生前后PAM-4信号的功率分布直方图.与现有的再生方案相比,本文方案的均匀多电平再生转移性能,使其更适合高频谱效率的长距空分复用系统和任意电平数的PAM信号再生.此外,该方案也能够扩展到波长域,有效提高光通信系统的传输容量.     

Facile synthesis of linear–hyperbranched polyphosphoesters via one‐pot tandem ROMP and ADMET polymerization and their transformation to architecturally defined nanoparticles

A convenient one‐pot synthesis of linear–hyperbranched polyphosphoesters (l–HBPPEs) was accomplished by a tandem ring‐opening metathesis polymerization (ROMP) and acyclic diene metathesis (ADMET) polymerization procedure. A linear monotelechelic poly(norbornene) with a terminal acrylate and many pendent thiol groups is first prepared through adding an internal cis‐olefin terminating agent to the reaction mixture immediately after the completion of the living ROMP, and then utilized as a macromolecular chain stopper in subsequent ADMET polymerization of a phosphoester functional AB₂ monomer, yielding l–HBPPEs as the reaction time prolonged. These l–HBPPEs bearing lots of pendent thiol groups in linear poly(norbornene) and peripheral acrylate groups in HBPPE could be self‐crosslinked in ultradilute solution through thiol‐Michael addition click reaction between acrylate and thiol to give single‐molecule nanoparticles with comparatively uniform size. This facile approach can be extended toward the fabrication of novel nanomaterials with sophisticated structures and tunable multifunctionalities. © 2015 Wiley Periodicals, Inc. J. Polym. Sci., Part A: Polym. Chem. 2015 , 53, 964–972