Combinational synthesis of oligonucleotides and assembly fabrication of oligonucleotide array

In this paper, a simple, reliable and flexible method, which integrated in situ synthesis with the spotting technique, was reported to fabricate oligonucleotide array. Different oligonucleotide sequences are synthesized on their relative code glass slides through combinational chemistry, thus the slides are broken into smaller pieces, in which the same code pieces have the same probe sequences. An oligonucleotide array is fabricated by arbitrarily assembling these different code pieces onto another solid substrate. In principle experimentation, four different sequences of P16 gene were synthesized and a 5 × 5 array including these four sequences and the control black was fabricated. The analysis results indicated that the hybridization fluorescence intensity of the same sequences locating different sets on the array gave the approximate values, and the fluorescence intensity ratio of matched sequence to one middle location base mismatched, two base mismatched, three middle base mismatched is (1.000 ± 0.080):(0.4991 ± 0.0671):(0.2360 ± 0.0044):(0.0493 ± 0.0033). Their relative accuracies were from 6.64 to 10.2%. This result might be used to rapidly screen single-nucleotide polymorphisms (SNPs).     

DNA microarray: a high throughput approach for methylation detection

We described a DNA microarray-based method combined with bisulphite treatment of DNA and regular PCR to examine hyper-methylation in promoter 1A of APC gene. A set of oligonucleotide probes were designed and immobilized on the aldehyde-coated glass slides for detecting the methylation pattern of 15 selected CpG sites in the region. The methylation status of 30 colorectal tumor samples have been examined by both of methylation-specific PCR (MS-PCR) and the present microarray method. The methylation pattern of the 15 CpG sites for the samples have been obtained with the microarray. A total of 19 samples out of 30 were methylated by microarray, in which five samples cannot be detected by MS-PCR due to the methylated CpG patterns not accordant to the MS-PCR primers. The detecting ratio for methylation of APC gene of colorectal tumor samples increased from 46.7% with MS-PCR to 63.3% with the microarray, which successfully demonstrated that DNA microarray-based method not only can obtained the methylation patterns for the related genes, but also decrease the false-negative results of methylation status by the conventional MS-PCR for the investigated genes.     

Four new triterpene glycosides from Thalictrum squarrosum

Four new triterpene glycosides were isolated from the dried aerial parts of Thalictrum squarrosum (Ranunculaceae). They were designated as squarroside I, being a cycloartane-type glycoside, and squarrosides II, III and IV, being oleanene-type glycosides. Their structures were established by using two dimensional (2D) NMR techniques.     

The first mu3-oxalato-bridged copper complex with tridentate Schiff base ligand N-ethyl-N'-salicylidene-1,2-diaminoethane: synthesis,structure, and magnetic properties

The first mu(3)-oxalato-bridged copper(II) complex, [[Cu(3)(L)(3)(mu(3)-C(2)O(4))][Cu(L)(H(2)O)](ClO(4))(2)] x 0.5(H(2)O) x 0.5(CH(3)OH), where HL = N-ethyl-N'-salicylidene-1,2-diaminoethane, has been synthesized and characterized by variable-temperature magnetic susceptibility measurement. The complex exhibits ferromagnetic coupling between the oxalato-bridged copper atoms and antiferromagnetic coupling between the oxygen-bridged copper atoms.     

Measuring intracellular calcium fluxes in high throughput mode

The measurement of intracellular calcium fluxes in real time is widely applied within the pharmaceutical industry to measure the activation of G-protein coupled receptors (GPCRhyp;s), either for pharmacological characterisation or to screen for new surrogate ligands. Initially restricted to G(q) coupled GPCRs, the introduction of promiscuous and chimeric G-proteins has further widened the application of these assays. The development of new calcium sensitive dyes and assays has provided sensitive, homogeneous assays which can be readily applied to high throughput screening (HTS). In this paper we describe the full automation of this assay type using a fluorometric imaging plate reader (FLIPR ) integrated into a Beckman/Sagian system to establish a simple robotic system that is well suited for the current medium throughput screening in this area of lead discovery. Using a recently completed HTS we discuss important determinants for FLIPR based screening, highlight some limitations of the current approach, and look at the requirements for future automated systems capable of keeping up with expanding compound files.     

Determination of proteins at nanogram levels by a resonance light-scattering technique with tetra-substituted sulphonated aluminum phthalocyanine

This is the first report on the determination of proteins with tetra-substituted sulphonated aluminum phthalocyanine (AlS₄Pc) by resonance light-scattering (RLS). At pH 3.0, the weak RLS of AlS₄Pc can be enhanced by the addition of proteins. Based on this, a novel quantitative method has been developed for the determination of proteins in aqueous solutions. Under optimal conditions, the linear ranges of the calibration curves were 0.050–2.0 μg ml⁻¹ for both human serum albumin (HSA) and human r-IgG. The detection limits were 12.7 ng ml⁻¹ for HSA and 16.1 ng ml⁻¹ for human r-IgG. The method has been applied to the analysis of total protein in human serum samples collected from the hospital and the results were in good agreement with those reported by the hospital, which indicates that the method presented here is not only sensitive, simple, but also reliable and suitable for practical applications.     

Stereoselective synthesis of substituted gamma-butyrolactones by the [3+2] annulation of allylic silanes with chlorosulfonyl isocyanate: enantioselective total synthesis of (+)-blastmycine

[structure: see text]. A stereoselective synthesis of gamma-butyrolactones by the [3 + 2] annulation of allylic silanes with N-chlorosulfonyl isocyanate (CSI) was developed. An enantioselective total synthesis of (+)-blastmycinone was accomplished using this annulation as the key step.     

带有控制-状态混合约束的时滞Volterra型积分系统的最优控制问题

对带有控制-状态混合约束的一类受控时滞Volterra型积分系统,导出了其最优控制的一组必要条件和一组充分条件．     

Rapid-phase modulation of terahertz radiation for high-speed terahertz imaging and spectroscopy

Rapid voltage-controlled phase modulation of cw terahertz (THz) radiation is demonstrated. By transmitting an infrared beam through a lithium niobate phase modulator the phase of the THz radiation, which is generated by the photomixing of two infrared beams, can be directly modulated through a 2pi phase shift. The 100 kHz modulation rate that is demonstrated with this technique is approximately 3 orders of magnitude faster than what can be achieved by mechanical scanning.     

The secondary structure of heated whey protein and its hydrolysates antigenicity

Fourier transform infrared spectroscopy(FTIR) and circular dichroism(CD) were used to investigate the conformational changes of heated whey protein(WP) and the corresponding changes in the hydrolysates immunoreactivity were determined by competitive enzyme-linked immunosorbent assay(ELISA).Results showed that the contents of α-helix and β-sheet of WP did not decrease much under mild heating conditions and the antigenicity was relatively high;when the heating intensity increased(70 ℃ for 25 min or 75 ℃ for 2...