Stress response induced by carbon nanoparticles in Paracentrotus lividus

Members of the 14-3-3 protein family are involved in many important cellular events, including stress response, survival and apoptosis. Genes of the 14-3-3 family are conserved from plants to humans, and some members are responsive to UV radiation. Despite the high rate of pollution generated by nano-pollutants, up to now their toxic effect on development is totally obscure. Embryos treated with carbon nanoparticles, RNA preparation, retro-transcription and quantitative real-time PCR. In response to carbon nano-particles exposure, the embryos collected 24 h later showed a 3,07-fold at 5x10(12) p and a 1,58-fold at 2.5x10(13) p and a 1,92-fold at 2.5x10(14) p increase in Pl14-3-3ε transcript levels compared with controls. The Pl14-3-3ε mRNA delocalization parallels the failure in archenteron elongation observed morphologically, as well as the lack of specific endoderm markers. Here, we report the isolation of the complete cDNA encoding the 14-3-3 epsilon isoform from Paracentrotus lividus sea urchin embryos, referred to as Pl14-3-3ε. Pl14-3-3ε mRNA levels were measured by RT-PCR during development and found to increase from the mesenchyme blastula to the prism stage. Our results confirm the involvement of 14-3-3ε in the stress response elicited by carbon nano-particles

Specific detection of topoisomerase I from the malaria causing P. falciparum parasite using isothermal rolling circle amplification

We present a Rolling-Circle-Enhance-Enzyme-Activity-Detection (REEAD) system with potential use for future point-of-care diagnosis of malaria. In the developed setup, specific detection of malaria parasites in crude blood samples is facilitated by the conversion of single Plasmodium falciparum topoisomerase I (pfTopI) mediated cleavage-ligation events, happening within nanometer dimensions, to micrometer-sized products readily detectable at the single molecule level in a fluorescence microscope. In principle, REEAD requires no special equipment and the readout is adaptable to simple colorimetric detection systems. Moreover, with regard to detection limit the presented setup is likely to outcompete standard gold immuno-based diagnostics. Hence, we believe the presented assay forms the basis for a new generation of easy-to-use diagnostic tools suitable for the malaria epidemic areas in developing countries