Assistance of maltose binding protein to the in vivo folding of the disulfide-rich C-terminal fragment from Plasmodium falciparum merozoite surface protein 1 expressed in Escherichia coli

The C-terminal fragment of Plasmodium falciparum merozoite surface protein 1 (F19) is a leading candidate for the development of a malaria vaccine. Successful vaccination trials on primates, immunochemistry, and structural studies have shown the importance of its native conformation for its protective role against infection. F19 is a disulfide-rich protein, and the correct pairing of its 12 half-cystines is required for the native state of the protein. F19 has been produced in the Escherichia coli periplasm, which has an oxidative environment favorable for the formation of disulfide bonds. F19 was either expressed as a fusion with the maltose binding protein (MBP) or directly addressed to the periplasm by fusing it with the MBP signal peptide. Direct expression of F19 in the periplasm led to a misfolded protein with a heterogeneous distribution of disulfide bridges. On the contrary, when produced as a fusion protein with E. coli MBP, the F19 moiety was natively folded. Indeed, after proteolysis of the fusion protein, the resulting F19 possesses the structural characteristics and the immunochemical reactivity of the analogous fragment produced either in baculovirus-infected insect cells or in yeast. These results demonstrate that the positive effect of MBP in assisting the folding of passenger proteins extends to the correct formation of disulfide bridges in vivo. Although proteins or protein fragments fused to MBP have been frequently expressed with success, our comparative study evidences for the first time the helping property of MBP in the oxidative folding of a disulfide-rich protein.     

Incorporación de la ecografía y la punción de tiroides a la actividad de endocrinología en una consulta de alta resolución

IntroductionFine-needle aspiration biopsy (FNAB) is considered the reference diagnostic procedure for thyroid nodules.Materials and methodsRoutine performance of thyroid ultrasound and ultrasound-guided FNAB by endocrinologists allows a more efficient approach in the setting of a high-resolution practice, thus reducing costs and the time elapsed until diagnosis.ResultsWe present our initial results of this procedure 2 years after its introduction, with a total of 286 biopsies performed. After a progressive learning curve over time and according to the endocrinologists’ previous experience, 72.72% samples were considered satisfactory for diagnosis. Greater difficulty was observed in obtaining optimal cytological specimens in smaller nodules. In conclusion, we have successfully incorporated thyroid ultrasound and ultrasound-guided FNAB into routine endocrine practice.ConclusionRoutine performance of thyroid ultrasound in endocrine practice will considerably aid the management of nodular thyroid disease.     

Wolbachia-host interactions: connecting phenotype to genotype

The long-established view of Wolbachia as reproductive parasites of insects is becoming complicated as an increasing number of papers describe a richer picture of Wolbachia-mediated phenotypes in insects. The search for the molecular basis for this phenotypic variability has been greatly aided by the recent sequencing of several Wolbachia genomes. These studies have revealed putative genes and pathways that are likely to be involved in the host-symbiont interaction. Whereas significant progress is being made from comparative genomic studies together with the use of model host systems like Drosophila, the ultimate linking of phenotype to genotype will require the development of genetic manipulation technology for both host and symbiont.     

Diversity and distribution of unicellular opisthokonts along the European coast analysed using high‐throughput sequencing

The opisthokonts are one of the major super groups of eukaryotes. It comprises two major clades: (i) the Metazoa and their unicellular relatives and (ii) the Fungi and their unicellular relatives. There is, however, little knowledge of the role of opisthokont microbes in many natural environments, especially among non‐metazoan and non‐fungal opisthokonts. Here, we begin to address this gap by analysing high‐throughput 18S rDNA and 18S rRNA sequencing data from different European coastal sites, sampled at different size fractions and depths. In particular, we analyse the diversity and abundance of choanoflagellates, filastereans, ichthyosporeans, nucleariids, corallochytreans and their related lineages. Our results show the great diversity of choanoflagellates in coastal waters as well as a relevant representation of the ichthyosporeans and the uncultured marine opisthokonts (MAOP). Furthermore, we describe a new lineage of marine fonticulids (MAFO) that appears to be abundant in sediments. Taken together, our work points to a greater potential ecological role for unicellular opisthokonts than previously appreciated in marine environments, both in water column and sediments, and also provides evidence of novel opisthokont phylogenetic lineages. This study highlights the importance of high‐throughput sequencing approaches to unravel the diversity and distribution of both known and novel eukaryotic lineages.     

Environmental assessment of the Atlantic mackerel (<Emphasis Type="Italic">Scomber scombrus</Emphasis>) season in the Basque Country. Increasing the timeline delimitation in fishery LCA studies

Purpose  

The purpose of this study was to evaluate the environmental impacts linked to fish extraction on a temporal basis, in order to analyze the effect that stock abundance variations may have on reporting environmental burdens. Inventory data for the North-East Atlantic Mackerel (NEAM) fishing season were collected over an 8-year period and used to carry out a life cycle assessment (LCA). The selected fishery corresponds to the Basque coastal purse seining fleet.     

Individual variation in behavioural plasticity: direct and indirect effects of boldness,exploration and sociability on habituation to predators in lizards

Little is known about the factors causing variation in behavioural plasticity and the interplay between personality and plasticity. Habituation to predators is a special case of behavioural plasticity. We investigated the direct and indirect effects of boldness, exploration and sociability traits on the habituation ability of Iberian wall lizards, considering exposure and sex effects. Individual boldness was consistent across several non-habituation contexts, but it did not significantly affect habituation. Exploration had a strong direct effect on habituation, with more exploratory individuals being able to habituate faster than less exploratory ones, probably because of their ability to assess risk better. Individual variation in habituation was also affected by sociability, but this was an indirect effect mediated by exposure to the predator. Less social individuals avoided refuges with conspecific cues, increasing exposure to the predator and eventually habituation. Finally, the direct effects of sex (females habituated faster than males) were opposite to its indirect effects through exposure. We conclude that risk assessment, instead of the proactivity–reactivity gradient usually considered in the literature, can affect behavioural plasticity through complex interactions between direct and indirect effects, including exploratory behaviour, degree of exposure to the predator and sex, which represent novel mechanisms generating inter-individual variation in plasticity.     

New names for old strains? <Emphasis Type="Italic">Wolbachia w</Emphasis>Sim is actually <Emphasis Type="Italic">w</Emphasis>Ri

A response to Serendipitous discovery of Wolbachia genomes in multiple Drosophila species by SL Salzberg, JC Dunning Hotopp, AL Delcher, M Pop, DR Smith, MB Eisen and WC Nelson. Genome Biology 2005, 6:R23