Pilot survey of expressed sequence tags (ESTs) from the asexual blood stages of Plasmodium vivax in human patients

Abstract Background Plasmodium vivax is the most widely distributed human malaria, responsible for 70–80 million clinical cases each year and large socio-economical burdens for countries such as Brazil where it is the most prevalent species. Unfortunately, due to the impossibility of growing this parasite in continuous in vitro culture, research on P. vivax remains largely neglected. Methods A pilot survey of expressed sequence tags (ESTs) from the asexual blood stages of P. vivax was performed. To do so, 1,184 clones from a cDNA library constructed with parasites obtained from 10 different human patients in the Brazilian Amazon were sequenced. Sequences were automatedly processed to remove contaminants and low quality reads. A total of 806 sequences with an average length of 586 bp met such criteria and their clustering revealed 666 distinct events. The consensus sequence of each cluster and the unique sequences of the singlets were used in similarity searches against different databases that included P. vivax, Plasmodium falciparum, Plasmodium yoelii, Plasmodium knowlesi, Apicomplexa and the GenBank non-redundant database. An E-value of <10-30 was used to define a significant database match. ESTs were manually assigned a gene ontology (GO) terminology Results A total of 769 ESTs could be assigned a putative identity based upon sequence similarity to known proteins in GenBank. Moreover, 292 ESTs were annotated and a GO terminology was assigned to 164 of them. Conclusion These are the first ESTs reported for P. vivax and, as such, they represent a valuable resource to assist in the annotation of the P. vivax genome currently being sequenced. Moreover, since the GC-content of the P. vivax genome is strikingly different from that of P. falciparum, these ESTs will help in the validation of gene predictions for P. vivax and to create a gene index of this malaria parasite.

Developmental basis of limb homology in Pleurodiran turtles, and the identity of the hooked element in the chelonian tarsus

Although Pleurodiran turtles represent an important component of extant turtle radiation, our knowledge of the development and homology of limb bones in turtles rests mostly upon observations made on derived members of the Cryptodiran clade. Herein, we describe limb development in three pleurodirans: Podocnemis unifilis Troschel, 1848, Podocnemis sextuberculata Cornalia, 1849 and Phrynops hilarii (Dumeril and Bibron, 1835), in an effort to contribute to filling this anatomical gap. For earlier stages of limb development, we described the Y-shaped condensation that gave rise to the zeugopodial cartilages, and differentiation of the primary axis/digital arch that reveals the invariant pattern common to tetrapods. There are up to four central cartilaginous foci in the carpus, and the proximal tarsale is formed by the fusion of the fibulare, intermedium, and centrale 4. Digital development is similar for the five digits. Changes in toe V occur predominantly in the distal tarsale 5. Ontogenetic reduction of phalanges is observed in toe V of Podocnemis. Based on these results, we suggest that the hooked element present in the chelonian tarsus, and traditionally recognized as a modified fifth metatarsale, is actually the fifth distal tarsale. Additionally, our data on limb development of pleurodiran turtles supply more taxonomically comprehensive information to interpret limb configuration within the chelonian clade. (C) 2009 The Linnean Society of London, Zoological Journal of the Linnean Society, 2009, 155, 845-866.