Minimizing 18O/16O back‐exchange in the relative quantification of ribonucleic acids

The use of isotopically labeled endonuclease digestion products allows for the relative quantification of ribonucleic acids (RNAs). This approach utilizes ribonucleases such as RNase T1 to mediate the incorporation of ¹⁸O onto the 3′‐terminus of the endonuclease digestion product from a solution containing heavy water (H₂¹⁸O). The accuracy and precision of relative quantification are dependent on the efficiency of isotope incorporation and minimizing any possible ¹⁸O to ¹⁶O back‐exchange before or during mass spectral analysis. Here, we have investigated the stability of ¹⁸O‐labeled endonuclease digestion products to back‐exchange. In particular, the effects of pH, temperature and presence of RNase on the back‐exchange process were examined using matrix‐assisted laser desorption/ionization mass spectrometry (MALDI‐MS). We have found that back‐exchange depends on the presence of the RNase—back‐exchange was not observed once the enzyme was removed from the sample. With RNase present, at all pH values examined (from acidic to basic pH), back‐exchange was detected at incubation above room temperature. The rates and extent of back‐exchange were similar at all pH values. In contrast, back‐exchange in the presence of RNase was found to be especially sensitive to incubation temperature—at temperatures below room temperature, minimal back‐exchange was detected. However, back‐exchange increased as the incubation temperature increased. Based on these findings, appropriate sample‐handling and sample storage conditions for isotopically labeled endonuclease digestion products have been identified, and these conditions should improve the accuracy and precision of results from the relative quantification of RNAs obtained by this approach. Copyright © 2009 John Wiley & Sons, Ltd.     

Hydrogen Bonding in Fluorinated Amides: FTIR,Two Dimensional Correlation Spectroscopy and DFT Calculations

Fluorinated macromers with amidic functional groups are used as additives in several high tech applications. We show here how aggregation phenomena related to hydrogen bonding are one of the key factor determining their chemical/physical and macroscopic properties. IR spectra are analyzed depending on different external parameters such as the concentration of amide groups and temperature. The experimental findings have been interpreted by means of DFT (Density Functional Theory) calculations on suitable molecular models. Moreover, 2D correlation spectroscopy has been applied to different sets of data, considering concentration and temperature as perturbing variables. The two dimensional correlation approaches confirmed the computational results and give an overall interpretation of the effects due to concentration and temperature.     

Existence of Solutions for Three Dimensional Stationary Incompressible Euler Equations with Nonvanishing Vorticity

In this paper, solutions with nonvanishing vorticity are established for the three dimensional stationary incompressible Euler equations on simply connected bounded three dimensional domains with smooth boundary. A class of additional boundary conditions for the vorticities are identified so that the solution is unique and stable.     

The analysis of a finite element method for the three-species Lotka–Volterra competition-diffusion with Dirichlet boundary conditions

A Galerkin finite element method is developed for the two dimensional/three dimensional nonlinear time-dependent three-species Lotka–Volterra competition-diffusion equations on a bounded domain. The existence and uniqueness of the solution to the numerical formulation are proved. An error estimate for the numerical solution is obtained. Numerical computations are carried out to examine the expected orders of accuracy in the error estimates.     

A meshless method for numerical solution of a linear hyperbolic equation with variable coefficients in two space dimensions

A meshless method is proposed for the numerical solution of the two space dimensional linear hyperbolic equation subject to appropriate initial and Dirichlet boundary conditions. The new developed scheme uses collocation points and approximates the solution employing thin plate splines radial basis functions. Numerical results are obtained for various cases involving variable, singular and constant coefficients, and are compared with analytical solutions to confirm the good accuracy of the presented scheme. © 2008 Wiley Periodicals, Inc. Numer Methods Partial Differential Eq, 2009     

Class field theory for a product of curves over a local field

We prove that the kernel of the reciprocity map for a product of curves over a p-adic field with split semi-stable reduction is divisible. We also consider the K ₁ of a product of curves over a number field.