酶促开环聚合与自缩合乙烯基共聚合相结合制取超支化聚合物

在Novozyme 435脂肪酶催化下, 甲基丙烯酸羟乙酯(HEMA)引发己内酯(ε-CL)开环聚合反应, 得到一端为双键, 另一端为羟基的直链聚己内酯(PCL)产物; 将其端羟基官能化得到大分子AB*型单体, 与苯乙烯以原子转移自由基聚合(ATRP)反应形式进行自缩合乙烯基共聚合, 得到超支化结构聚苯乙烯-b-聚己内酯产物.     

Role of binding entropy in the refinement of protein-ligand docking predictions: analysis based on the use of 11 scoring functions

We present results of testing the ability of eleven popular scoring functions to predict native docked positions using a recently developed method (Ruvinsky and Kozintsev, J Comput Chem 2005, 26, 1089) for estimation the entropy contributions of relative motions to protein-ligand binding affinity. The method is based on the integration of the configurational integral over clusters obtained from multiple docked positions. We use a test set of 100 PDB protein-ligand complexes and ensembles of 101 docked positions generated by (Wang et al. J Med Chem 2003, 46, 2287) for each ligand in the test set. To test the suggested method we compared the averaged root-mean square deviations (RMSD) of the top-scored ligand docked positions, accounting and not accounting for entropy contributions, relative to the experimentally determined positions. We demonstrate that the method increases docking accuracy by 10-21% when used in conjunction with the AutoDock scoring function, by 2-25% with G-Score, by 7-41% with D-Score, by 0-8% with LigScore, by 1-6% with PLP, by 0-12% with LUDI, by 2-8% with F-Score, by 7-29% with ChemScore, by 0-9% with X-Score, by 2-19% with PMF, and by 1-7% with DrugScore. We also compared the performance of the suggested method with the method based on ranking by cluster occupancy only. We analyze how the choice of a clustering-RMSD and a low bound of dense clusters impacts on docking accuracy of the scoring methods. We derive optimal intervals of the clustering-RMSD for 11 scoring functions.     

ICP-MS测量环境样品中铀的非质谱干扰内标校正研究

内标的合理应用直接关系到测量结果的可靠性,通过外标与内标校正相结合的方法,实验研究了Y,Rh,In,Tb,Yb,Tl,Bi,Re作为内标对ICP-MS测量铀的非质谱干扰的校正效果。对于非抑制/增强基体效应,各内标元素校正结果明显优于无内标校正结果,高质量数内标元素Tb,Yb,Re,Tl,Bi可获得较好的校正效果,校正偏差优于5%,其中以Tl为最佳;对于盐度较高的样品溶液,由于基体对待测元素和内标元素产生了明显不同的抑制/增强效应,因此各内标校正结果均不理想,这表明内标对可溶性固体引起的抑制/增强效应的校正能力有限。在铀的测量中,对于含盐度在300～500μg/mL的样品,内标校正造成的偏差可达到5%～10%;对于盐度在500μg/mL以上的样品,内标校正造成的偏差很可能超过10%或者更高。     

Tripodal borate ligands from tris(dimethylamino)borane: the first synthesis of a chiral tris(methimazolyl)borate ligand, and the crystal structure of a single diastereomer pseudo-C3-symmetric Ru(II) complex

The activation of tris(dimethylamino)borane towards reaction with a chiral methimazole by N-methylimidazole has been used to prepare the first example of a chiral tris(methimazolyl)borate ligand. Coordination of this neutral ligand to Ru(II) has been achieved by reaction with [(p-cymene)RuCl(2)](2) to provide a single diastereomer complex in which the chirality of the methimazolyl substituents dictate the chirality of the bicyclo[3.3.3]cage formed by the ligand on coordination to the metal. The alternative approach to chiral tris(methimazolyl)borate ligands involving the introduction of a chiral group onto the boron atom has been explored by replacing N-methylimidazole in the above reaction by chiral oxazolines as activating bases in reaction with simple methimazole. However, although the B(NMe(2))(3) is activated to reaction with methimazole by these oxazolines, an intramolecular oxazoline ring-opening by a coordinated methimazolyl sulfur occurs and prevents the successful synthesis of these ligands.     

Improved short-lived nuclide activation analysis

A novel technique has been developed to increase the sensitivity, precision and accuracy of short-lived nuclide activation analysis and especially of delayed fission neutron counting, by improving the counting statistics. This has been achieved by approaching the irradiated sample to the detector during the counting period in order to compensate for the rapid radioactive decay by variation of the counting efficiency, permitting prolongation of the counting period with constant count rate. The sample holder movement with approximately exponentially decreasing velocity is implemented by a variable DC motor with a rotary to linear movement converter, which is being replaced by a more reliable stepping motor.     

Enhanced melanin fluorescence by stepwise three-photon excitation

The fluorescence of eumelanin (from Sepia officinalis and black human hair) was activated and enhanced by almost three orders of magnitude by exposure to near-infrared radiation. No activation or enhanced emission was observed when the samples were heated up to 100°C. The near-infrared irradiation caused obvious changes to the eumelanin and could be seen by fluorescence and bright field imaging. The area of enhanced emission appeared to originate from a region with changes in the morphology of the eumelanin's granule and increased with exposure time. At least two different components with enhanced fluorescence were activated and could be distinguished by their excitation properties. One component could be excited efficiently with wavelengths in the visible region and exhibited linear absorption dependence with respect to the laser power level. The second component could be excited efficiently using near-infrared wavelengths by a nonlinear process and exhibited a third-order dependence on the excitation. The third-order dependence is explained by a step-wise excited-state absorption process since the same third-order dependence was present when either continuous wave or femtosecond pulsed laser, with similar average-power levels, was used.     

四环素-铜(Ⅱ)配合物与DNA相互作用的吸收光谱研究

用紫外光谱方法研究了四环素(TC) Cu(II)配合物与DNA的相互作用.吸收光谱研究表明,DNA能与四环素(TC)及Cu(II)形成的配合物发生反应,配合物与DNA的作用方式随着配合物类型及DNA浓度的不同而不尽相同:当四环素与铜形成1∶1型配合物时,较低浓度的DNA能与配合物以嵌插方式相互作用,而较高浓度的DNA与该配合物除了发生嵌插作用外,还存在另外的作用方式;当四环素与铜形成1∶2型配合物时,DNA与该配合物则主要以嵌插方式相互作用,并且这两种配合物与DNA的嵌插作用均是通过四环素配体插入的.     

Spot-test detection of aryl hydrazines, hydrazones, osazones, oximes and aromatic amines

Arylhydrazines are detected by oxidation with N-bromosuccinimide and coupling with resorcinol to form azo-dyes which are intensely coloured in alkaline media. Hydrazones and osazones are hydrolysed to form the arylhydrazines, which are then tested for 4-Nitro- and 2,4-dinitrophenylhydrazines are tested for by forming their hydrazone with salicylaldehyde and adding alkali to produce a violet colour. The hydroxylamine formed by the hydrolysis of oximes is oxidized by iodine monochloride in the presence of sulphanilic acid; coupling with 8-hydroxyquinoline forms a dye that is red in alkali. Intense colours are immediately produced when primary, secondary and tertiary aromatic amines are mixed with diacetoxyiodobenzene. All the tests are sensitive and appear to be specific.     

Calixarenes with nitrogen or phosphorus substituents on the lower rim

Calix[n]arenes (n=4,6) with diphenylphosphinite groups appended to their lower rim have been synthesized by reaction first with base, followed by chlorodiphenylphosphine. The reaction has also been carried out with the partially methoxylated calix[n]arenes. Calix[6]arenes with phosphate groups selectively bridging adjacent pairs of oxygens have been synthesized by reaction first with base, followed by ethyl dichlorophosphate. Calix[n]arenes (n=4,6) with 2-aminoethyloxy groups appended to the lower rim have been synthesized both by the reduction of an amide or nitrile group. Calixarenes with 2-hydroxyethyloxy and 2-bromoxyethyloxy groups appended to the lower rim have also been prepared. A route to preparing calixarene-functionalized polymers by the alkylation of polyethyleneimine is also described.This paper is dedicated to the commemorative issue on the 50th anniversary of calixarenes.     

A minimal multiconfigurational technique

A direct minimization method previously presented by the authors is applied here to biconfigurational wave functions. A very moderate increasing in the time by iteration with respect to the one-determinant calculation and good convergence properties have been found. So qualitatively correct studies on singlet systems with strong biradical character can be performed with a cost similar to that required by Hartree-Fock calculations.     

Sulfines and Sulfenes – the S-Oxides and S,S-Dioxides of Thioaldehydes and Thioketones

Four sulfines have so far been prepared by elimination of HCl from sulfinyl chlorides with triethylamine and by oxidation of diaryl thioketones with peracid. They are colored, crystalline compounds, which slowly decompose at room temperature. Attempts to prepare sulfenes have not yet been successful, but mesylsulfene^[1] has been isolated as the trimethylamine adduct. It can be shown by interception reactions that sulfenes are formed as intermediates by the action of trialkylamines on aliphatic sulfonyl chlorides and by the action of sulfur dioxide on diazoalkanes. They react with ROD to form monodeuterated sulfonates, with diazoalkanes to give three-membered rings, with enamines, dienamines, ketene O,O,O,N-, and N,N-acetals, and chloral, and in some cases with vinyl ethers, to give four-membered rings. They also react with nitrones to give five-membered rings, and with β-aminovinyl ketones and dienamines to give six-membered rings.     

Capillary zone electrophoresis for the quantitation of oligosaccharides formed through the action of chitinase.

Capillary zone electrophoresis with fluorescence detection was used to analyze the products formed by chitinase acting on N-acetylchitooligosaccharide-fluorescent conjugates. Six oligosaccharides of the structure [N-acetylglucosamine(1----4)]n (where n = 1-6) were conjugated to 7-amino-1,3-naphthalene disulfonic acid by reductive amination. Each oligosaccharide-fluorescent conjugate was purified by preparative gradient polyacrylamide gel electrophoresis, semi-dry electrotransfer to a positively-charged nylon membrane and recovered by washing the membrane with salt solution. The products formed by treating each oligosaccharide-fluorescent conjugate with chitinase were analyzed by capillary zone electrophoresis. The chitinase treatment hexasaccharide-fluorescent conjugate was also examined kinetically to study the action pattern of this enzyme.     

Highly efficient proteome analysis with combination of protein pre-fractionation by preparative microscale solution isoelectric focusing and identification by μRPLC-MS/MS with serially coupled long microcolumn

To improve the efficiency of proteome analysis, a strategy with the combination of protein pre-fractionation by preparative microscale solution isoelectric focusing, peptide separation by μRPLC with serially coupled long microcolumn and protein identification by ESI-MS/MS was proposed. By preparative microscale solution isoelectric focusing technique, proteins extracted from whole cell lysates of Escherichia coli were fractionated into five chambers divided by isoelectric membranes, respectively with pH range from 3.0 to 4.6, 4.6 to 5.4, 5.4 to 6.2, 6.2 to 7.0 and 7.0 to 10.0. Compared to the traditional on-gel IFF, the protein recovery could be obviously improved to over 95%. Subsequently, the enriched and fractionated proteins in each chamber were digested, and further separated by a 30-cm long serially coupled RP microcolumn. Through the detection by ESI-MS/MS, about 200 proteins were identified in each fraction, and in total 835 proteins were identified even with one-dimensional μRPLC-MS/MS system. All these results demonstrate that by such a combination strategy, highly efficient proteome analysis could be achieved, not only due to the in-solution protein enrichment and pre-fractionation with improved protein recovery but also owing to the increased separation capacity of serially coupled long μRPLC columns.     

Water-soluble nanodiamond

Reduction of the graphenic edges of annealed nanodiamond by sodium in liquid ammonia leads to a nanodiamond salt that reacts with either alkyl or aryl halides by electron transfer to yield radical anions that dissociate spontaneously into free radicals and halide. The free radicals were observed to add readily to the aromatic rings of the annealed nanodiamond. Nanodiamonds functionalized by phenyl radicals were sulfonated in oleum, and the resulting sulfonic acid was converted to the sodium salt by treatment with sodium hydroxide. The solubility of the salt in water was determined to be 248 mg/L. Nanodiamond functionalized by carboxylic acid groups could be prepared by reacting 5-bromovaleric acid with the annealed nanodiamond salt. The solubility of the sodium carboxylate in water was found to be 160 mg/L.     

Mass spectrometry of 4-amino-4′-nitroazobenzene compounds

The mass spectra of 32 substituted 4-amino-4′-nitroazobenzene compounds have been recorded and the most intense peaks have been used to characterize these spectra. It was found that the spectra of 4-amino-4′-nitroazobenzene compounds are characterized by peaks due to: (1) molecular ions, (2) fragment ions formed by cleavage of one of the carbon-nitrogen bonds adjacent to the azo linkage with the positive charge remaining with the amine fragment, (3) ions formed by cleavage alpha to the amine nitrogen with the charge remaining with the amine substituent, (4) ions formed by cleavage beta to the amine nitrogen with the loss of the amine substituent fragment, (5) secondary ions formed by cleavage beta to the amine nitrogen with the loss of the amine substituent fragment from the primary amine fragment (2), and (6) ions formed by loss of NO from the molecular ion. This work shows that 4-amino-4′-nitroazobenzene compounds exhibit fragmentation which is dependent in a consistent manner on the types of substituents. This work provides a basis for a systematic approach to the identification of 4-amino-4′-nitroazobenzene compounds.     

Enhancement of gemcitabine affinity for biomembranes by conjugation with squalene: differential scanning calorimetry and Langmuir-Blodgett studies using biomembrane models

Molecular interactions between gemcitabine, alone or conjugated with squalene to form the gem-squalene prodrug, with dimyristoylphosphatidylcholine have been investigated by differential scanning calorimetry and Langmuir film balance techniques to gain information about the interaction of gemcitabine and its prodrug with mammalian cell membranes and to evaluate the potential of liposomes as a delivery system for gemcitabine prodrugs. Phospholipids assembled as multilamellar vesicles or monolayers (at the air water interface) have been used as biomembrane models. Different interactions of gemcitabine, its prodrug, and squalene with the lipid were detected by dispersing the compounds in the MLV and were compared with kinetic experiments carried out to consider the ability of the examined compounds to dissolve in an aqueous medium, to migrate through it, and to be captured by multilamellar vesicles. Their ability to be released from drug-loaded liposomes and be taken up by empty vesicles mimicking biomembranes was also considered. Analysis of the differential scanning calorimetry curves reveals that gemcitabine has very little interaction with multilamellar vesicles whereas the gem-squalene prodrug strongly interacts with multilamellar vesicles. The kinetic experiments suggest that an aqueous medium does not permit the prodrug uptake by the biomembrane models, whereas it is allowed when gem-squalene is gradually released by the liposomes. The molecular area/surface pressure isotherms of the gemcitabine/lipid, gem-squalene/lipid, and pure compound monolayers, in agreement with the calorimetric results, indicate that gem-squalene interacts with the phospholipid monolayer with the squalene moiety in contact with the phospholipid chains and gemcitabine protruding in the aqueous medium.     

Determination of fatty acid methyl esters: Elimination of tissue-derived contamination and artefacts and preparation of a support-coated acid-modified polyester liquid phase

Fatty acid methyl esters made by transesterification of extracted plasma lipids with methanolic hydrochloric acid were found to contain substances which interfered with quantitation by gas-liquid chromatography. A method to eliminate most interfering materials and a procedure that allows reproducible wetting of an inert support with acid-modified ethylene glycol succinate are reported. Hydrocarbon contamination and artefacts produced during transesterification from substances native to tissue matrices were identified by gas chromatography/mass spectrometry and removed by chromatography over 6% hydrated alumina. Ethylene glycol succinate was treated with orthophosphoric acid to improve wetting of silanized supports and extend the thermal stability of the polymer. Quantitation was achieved by using synthetic triheptadecanoin as an internal standard.     

Triphenyltelluroniumthiolate

Triphenyltelluroniumthiolates Triphenyltelluroniumthiolates are prepared by reaction of Triphenyltelluroniumchloride with Sodiumthiolates or by reaction of Triphenyltelluroniummethanolates with Thiols. The title compounds decompose by heating to Diphenyltelluride and Phenylthioether. Reaction of the Triphenyltelluroniumalkylthiolates with CS₂ leads to the Triphenyltelluroniumalkylthioxanthates.     

Analysis of chicken and turkey ovalbumins by microchip electrophoresis combined with exoglycosidase digestion

The polypeptide and carbohydrate patterns of two glycoproteins, chicken ovalbumin (CO) and turkey ovalbumin (TO), were analyzed by microchip electrophoresis (ME), following digestion with proteases and exoglycosidases. Glycopeptides derived from ovalbumin were obtained by digestion with Pronase, followed by dialysis, and then separated by ME. Using CO as model, the method was developed to deduce the structure of glycans from glycoproteins by comparing the electropherograms of glycopeptides with and without digestion of exolycosidases. Applying the same approach, the structure of oligosaccharides linked to TO was determined. TO was found to contain high-mannose type oligosaccharides and oligosaccharides with terminal N-acetylglucosamine residues. The complete primary analysis of CO and TO by ME described in this paper provides a basis for an analysis of glycoproteins with an integrated microfluidic chip.     

Zur Analyse der Sulfobetaine

The analysis of the sulphobetaines is dealt with with special attention to the separation from all other detergents. Contrary to other ampholytic detergents the sulphobetaines are recovered together with the nonionic detergents by using ion-exchange methods. They can be separated from the non-ionic detergents by a thin-layer chromatographic method. The IR- and NMR-spectra allowing the identification are discussed. The quantitative determination of the sulphobetaines cannot be accomplished by means of the methods normally used in detergent analysis. It has to be done with the aid of chemical decomposing reactions by a determination of sulphur.