Optimal kinetics for quantification of antigen-induced cytokines in human peripheral blood mononuclear cells by real-time PCR and by ELISA.

Real-time polymerase chain reaction (PCR) has recently been described as a new tool to measure and accurately quantify mRNA levels. In this study, we have applied this technique to evaluate cytokine mRNA synthesis induced by antigenic stimulation with purified protein derivative (PPD) or heparin-binding haemagglutinin (HBHA) in human peripheral blood mononuclear cells (PBMC) from Mycobacterium tuberculosis-infected individuals. Whereas PPD and HBHA optimally induced IL-2 mRNA after respectively 8 and 16 to 24 h of in vitro stimulation, longer in vitro stimulation times were necessary for optimal induction of interferon-gamma (IFN-gamma) mRNA, respectively 16 to 24 h for PPD and 24 to 96 h for HBHA. IL-13 mRNA was optimally induced by in vitro stimulation after 16-48 h for PPD and after 48 to 96 h for HBHA. Comparison of antigen-induced Th1 and Th2 cytokines appears, therefore, valuable only if both cytokine types are analysed at their optimal time point of production, which, for a given cytokine, may differ for each antigen tested. Results obtained by real-time PCR for IFN-gamma and IL-13 mRNA correlated well with those obtained by measuring the cytokine concentrations in cell culture supernatants, provided they were high enough to be detected. We conclude that real-time PCR can be successfully applied to the quantification of antigen-induced cytokine mRNA and to the evaluation of the Th1/Th2 balance, only if the kinetics of cytokine mRNA appearance are taken into account and evaluated for each cytokine measured and each antigen analysed.

PCR from the CPR offers a historical perspective on marine population ecology.

The Continuous Plankton Recorder (CPR) survey has collected plankton samples from regular tracks across the world's oceans for almost 70 y. Over 299,000 spatially extensive CPR samples are archived and stored in buffered formalin. This CPR archive offers huge potential to study changes in marine communities using molecular data from a period when marine pollution, exploitation and global anthropogenic impact were much less pronounced. However, to harness the amount of data available within the CPR archive fully, it is necessary to improve techniques of larval identification, to genus and species preferably, and to obtain genetic information for historical studies of population ecology. To increase the potential of the CPR database this paper describes the first extraction, amplification by the polymerase chain reaction and utilization of a DNA sequence (mitochondrial 16S rDNA) from a CPR sample, a formalin fixed larval sandeel.

Full-Wave Analysis of Layered Aperture Arrays

The new rigorous numerical-analytical technique based upon Galerkin method with the entire domain basis functions has been developed and applied to the study of the periodic aperture arrays containing multiple dissimilar apertures of complex shapes in stratified medium. The rapid uniform convergence of the solutions has enabled a comprehensive parametric study of complex array arrangements. The developed theory has revealed new effects of the aperture shape and layout on the array performance. The physical mechanisms underlying the TM wave resonances and Luebbers' anomaly have been explained for the first time.