Expression of preproinsulin-2 gene shapes the immune response to preproinsulin in normal mice

Deciphering mechanisms involved in failure of self tolerance to preproinsulin-2 is a key issue in type 1 diabetes. We used nonautoimmune 129SV/Pas mice lacking preproinsulin-2 to study the immune response to preproinsulin-2. In these mice, a T cell response was detected after immunization with several preproinsulin-2 peptides and confirmed by generating hybridomas. Activation of some of these hybridomas by wild-type (wt) islet cells or recombinant murine proinsulin-2 demonstrated that two epitopes can be generated from the naturally expressed protein. Although T cells from wt mice responded to preproinsulin-2 peptides, we could not detect a response to the naturally processed epitopes in these mice. Moreover, after immunization with recombinant whole proinsulin-2, a T cell response was detected in preproinsulin-2-deficient but not in wt mice. This suggests that islet preproinsulin-2-autoreactive T cells are functionally eliminated in wt mice. We used a transplantation model to evaluate the relevance of reactivity to preproinsulin-2 in vivo. Wild-type preproinsulin-2-expressing islets transplanted in preproinsulin-2-deficient mice elicited a mononuclear cell infiltration and insulin Abs. Graft infiltration was further increased by immunization with preproinsulin-2 peptides. Preproinsulin-2 expression thus shapes the immune response and prevents self reactivity to the islet. Moreover, islet preproinsulin-2 primes an immune response to preproinsulin-2 in deficient mice.     

Coenzyme based synthesis of silver nanocrystals

In this work we have carried out systematic studies to identify the critical role of a coenzyme (β-NADPH) to synthesize silver nanoparticle. Interestingly, both roles of reducing and stabilizing agents are played by β-NADPH. Nanoparticles obtained by this route exhibit a good crystallinity, a narrow size distribution and excellent stability in aqueous solution. The most advantageous points of this single-step environmentally friendly approach are that it takes place at nearly room temperature (20°C), overcomes many limitations encountered in other biological methods (such as the restricted concentration of AgNO(3), maintenance and manipulation of microorganisms, preparing extracts and contamination from residual reactants), bypasses the use of surfactants or capping agents and does not necessitate pH adjustment. The nano-Ag were characterized using transmission electron microscopy (TEM), X-ray diffraction (XRD), dynamic light scattering (DLS), zeta potential, UV-vis, and energy-dispersive X-ray spectroscopy (EDX). DLS, TEM and XRD measurements showed the formation of nano-Ag with an average diameter of 20.77±0.67nm. XRD studies confirmed the nanocrystalline nature of the silver particles. Zeta potential measurements revealed that the particles are surrounded with negatively charged groups (-41±5mV) making them stable in an aqueous medium. The EDX spectrum of the silver nanoparticles confirmed the presence of elemental silver signal in high percentage. In addition to the easy and ecofriendly method of synthesis, β-NADPH can be regenerated by enzymatic means through glucose 6-phosphate dehydrogenase, potentially making the synthesis more cost effective.     

Fertilization and wounding of the style induce the expression of a highly conserved plant gene homologous to a Plasmodium falciparum surface antigen in the wild potato Solanum chacoense Bitt.

Pistil tissues are actively involved in pollen tube growth and respond to the presence of the growing pollen tubes by modulating the expression of specific genes. Once fertilization has occurred, complex developmental programs lead to embryogenesis, ovary maturation, and seed set. In order to understand the early events that follow pollination and fertilization we have used a subtractive hybridization approach to characterize genes which are related to pollination and fertilization events. One cDNA clone isolated and named SPP30 (Solanum pollinated pistil) was found to share significant sequence identities with a Plasmodium falciparum (malaria parasite) surface antigen and a yeast gene of unknown function. Searches in recent EST databases also revealed that SPP30 homologues are found in both monocot and dicot species. The presence of this conserved gene in evolutionarily distant organisms such as yeast, Plasmodium, and plants suggests that it codes for an essential cellular function. This is also strengthened by its extremely high sequence conservation in both monocots and dicots where virtually all substitutions tolerated are conservative.     

Identification,characterization, and regulation of a cluster of genes involved in carbapenem biosynthesis in Photorhabdus luminescens

The luminescent entomopathogenic bacterium Photorhabdus luminescens produces several yet-uncharacterized broad-spectrum antibiotics. We report the identification and characterization of a cluster of eight genes (named cpmA to cpmH) responsible for the production of a carbapenem-like antibiotic in strain TT01 of P. luminescens. The cpm cluster differs in several crucial aspects from other car operons. The level of cpm mRNA peaks during exponential phase and is regulated by a Rap/Hor homolog identified in the P. luminescens genome. Marker-exchange mutagenesis of this gene in the entomopathogen decreased antibiotic production. The luxS-like signaling mechanism of quorum sensing also plays a role in the regulation of the cpm operon. Indeed, luxS, which is involved in the production of a newly identified autoinducer, is responsible for repression of cpm gene expression at the end of the exponential growth phase. The importance of this carbapenem production in the ecology of P. luminescens is discussed.