Microchip electrophoresis for specific gene detection of the pathogenic bacteria V. cholerae by circle-to-circle amplification.

We have developed a new method for a fast and precise analysis of circle-to-circle amplification (C2CA) product for specific gene detection by microchip electrophoresis. In this method, we have added a new enzymatic step to the C2CA reaction, which could be carried out isothermally at 37 degrees C. Compared to the original single-stranded DNA, the double-stranded DNA that is produced by this enzymatic reaction is more reliable for analysis by microchip electrophoresis. C2CA product was detected within 55 s with high reproducibility by this method which was successfully applied to the detection of 10-ng genomic DNA of the pathogenic bacteria Vibrio. cholerae within 110 s. Purification was found to be an indispensable step for the analysis of the C2CA product of genomic DNA samples.     

Thermal decomposition of poly(sec-amyl methacrylate)

The thermal decomposition of poly(sec-amyl methacrylate) is studied by simultaneous thermogravimetry–gas chromatography–mass spectrometry and by pyrolysis–gas chromatography. The TG curve has four distinct breaks and a plateau. Results of the identification of the evolved gas at the individual breaks by GC–MS techniques lead to the conclusion that these breaks correspond to the individual processes in the decomposition mechanism like that of poly(tert-butyl methacrylate): the first break, the depolymerization initiated at the unsaturated chain ends; the second break, the depolymerization initiated at the saturated chain ends; the third break, the ester decomposition; the plateau, the inhibition of decomposition by the formation of poly(methacrylic anhydride); the fourth break, the decomposition of poly(methacrylic anhydride). The extent of ester decomposition is related to the substituent constants based on Hammett equation. The ester decomposition product is separated into three pentene isomers by pyrolysis–gas chromatography: trans-2-pentene, cis-2-pentene, and 1-pentene. As raising decomposition temperature, the composition ratio of trans-2-pentene decreases and becomes constant above 620 K, and the composition ratios of cis-2-pentene and 1-pentene increase and also become constant above 620 K. These results are accounted for by mobility of atoms included in the substituent at forming a ring transition state.     

Chemical shifts in methylcyclohexane

In order to determine the effect of a single methyl group on the chemical shifts of protons in a cyclohexane ring, methylcyclohexane-1,2,3,3,5,5,-d₆ has been synthesized. The protons in the 4-position and the 2,6-equatorial protons are not significantly different from those in ring-frozen cyclohexane. The 2,6-axial protons, however, experience an up-field shift of about 15 Hz from the position of the other axial protons. These observations are discussed in terms of the structure of methylcyclohexane.     

Treatment of ion exchange membranes to decrease divalent ion permeability

In the electrodialysis process for concentrating sea water, the addition of a small amount of the polycationic reagent Nonisold into the sea water feed results in a reduction of divalent cation permeability relative to that of monovalent cations. This is due to the formation of a reagent layer on the surface of the cation-exchange membrane. Divalent cations require greater energy to pass over the potential barrier at the reagent layer than do monovalent cations. The relationship between the height of the potential barrier and the ratio of ionic fluxes or the permselectivity coefficient is deduced on the basis of a kinetic-controlled mechanism of uptake of ions from the solution, and the height of the potential barrier is estimated from the experimental results. The difference between the potential barrier for divalent cations and that for monovalent cations may reach up to about 10 kJ/mol.     

Practical and convenient synthesis of N-heterocycles: stereoselective cyclization of N-alkenylamides with t-BuOI under neutral conditions

[Structure: see text] tert-Butyl hypoiodite (t-BuOI) was found to be a powerful reagent for the cyclization of N-alkenylamides leading to a variety of N-heterocycles under extremely mild conditions. When N-alkenylsulfonamides were employed in the reaction, three- to six-membered saturated N-heterocycles were obtained in good to excellent yields with complete stereoselectivity. The method was applicable to the cyclization of alkenylbenzamide derivatives to afford N-, O- or N-, S-heterocycles.     

Nonlinear viscoelastic properties and change in entanglement structure of linear polymer

Simultaneous measurements of stress relaxation and differential dynamic modulus were made at 268 K over a time scale of 10 to 10⁴⁵ s for nearly monodisperse polybutadiene (M _w =2.2x10⁵, 1,2-structure 70%, M _e =3600) and also one having coarse cross-linking (M _c =29000). Static shear strain ranged from 0.1 to 2.0. In a long-time region (t> _k ), the relaxation modulus G (; t) could be expressed by the product G (0; t) h (y). The observed h() agreed well with the Doi-Edwards theory without use of IA approximation. Both the cured and uncured samples showed initial drop of the differential storage modulus G (), ; t) followed by gradual recovery, but did not attain the value before shearing G (, ; t) for the uncured sample showed smaller values than that for the cured one in the whole measured time scale at the higher strain, confirming the two origins of nonlinear viscoelasticity of well entangled polymer; induced chain anisotropy and induced decrement in entanglement density. G (, ; t) curves for the cured sample agreed well with the BKZ predictions. But the curves for the uncured sample agreed well with the BKZ prediction only at the time scale of t< _k . BKZ prediction showed significant upward deviations at t> _k . Such the differences are discussed in terms of the two origins.Dedicated to Prof. John D. Ferry on the occasion of his 85th birthday.     

Nuclease tolerant FRET probe based on DNA-quantum dot conjugation.

We have developed a fluorescence resonance energy transfer (FRET) probe based on the conjugation of a quantum dot (QD) with dye (YOYO-3) intercalated DNA. The FRET-inducing electrostatic coupling of DNA and the QD made structural changes to the QD-DNA conjugates, which significantly prevented an enzymatic reaction between the DNA and a conventional restriction endonuclease (EcoRI).