An overview of malaria transmission from the perspective of Amazon Anopheles vectors

In the Americas, areas with a high risk of malaria transmission are mainly located in the Amazon Forest, which extends across nine countries. One keystone step to understanding the Plasmodium life cycle in Anopheles species from the Amazon Region is to obtain experimentally infected mosquito vectors. Several attempts to colonise Ano- pheles species have been conducted, but with only short-lived success or no success at all. In this review, we review the literature on malaria transmission from the perspective of its Amazon vectors. Currently, it is possible to develop experimental Plasmodium vivax infection of the colonised and field-captured vectors in laboratories located close to Amazonian endemic areas. We are also reviewing studies related to the immune response to P. vivax infection of Anopheles aquasalis, a coastal mosquito species. Finally, we discuss the importance of the modulation of Plasmodium infection by the vector microbiota and also consider the anopheline genomes. The establishment of experimental mosquito infections with Plasmodium falciparum, Plasmodium yoelii and Plasmodium berghei parasites that could provide interesting models for studying malaria in the Amazonian scenario is important. Understanding the molecular mechanisms involved in the development of the parasites in New World vectors is crucial in order to better determine the interaction process and vectorial competence.

Computational drug design strategies applied to the modelling of human immunodeficiency virus-1 reverse transcriptase inhibitors

Reverse transcriptase (RT) is a multifunctional enzyme in the human immunodeficiency virus (HIV)-1 life cycle and represents a primary target for drug discovery efforts against HIV-1 infection. Two classes of RT inhibitors, the nucleoside RT inhibitors (NRTIs) and the nonnucleoside transcriptase inhibitors are prominently used in the highly active antiretroviral therapy in combination with other anti-HIV drugs. However, the rapid emergence of drug-resistant viral strains has limited the successful rate of the anti-HIV agents. Computational methods are a significant part of the drug design process and indispensable to study drug resistance. In this review, recent advances in computer-aided drug design for the rational design of new compounds against HIV-1 RT using methods such as molecular docking, molecular dynamics, free energy calculations, quantitative structure-activity relationships, pharmacophore modelling and absorption, distribution, metabolism, excretion and toxicity prediction are discussed. Successful applications of these methodologies are also highlighted.

Identification of the nicotinamide mononucleotide adenylyltransferase of Trypanosoma cruzi

The intracellular parasite Trypanosoma cruzi is the aetiological agent of Chagas disease, a public health concern with an increasing incidence rate. This increase is due, among other reasons, to the parasite’s drug resistance mechanisms, which require nicotinamide adenine dinucleotide (NAD+). Furthermore, this molecule is involved in metabolic and intracellular signalling processes necessary for the survival of T. cruzi throughout its life cycle. NAD+ biosynthesis is performed by de novo and salvage pathways, which converge on the step that is catalysed by the enzyme nicotinamide mononucleotide adenylyltransferase (NMNAT) (enzyme commission number: 2.7.7.1). The identification of the NMNAT of T. cruzi is important for the development of future therapeutic strategies to treat Chagas disease. In this study, a hypothetical open reading frame (ORF) for NMNAT was identified in the genome of T. cruzi. The corresponding putative protein was analysed by simulating structural models. The ORF was amplified from genomic DNA by polymerase chain reaction and was further used for the construction of a corresponding recombinant expression vector. The expressed recombinant protein was partially purified and its activity was evaluated using enzymatic assays. These results comprise the first identification of an NMNAT in T. cruzi using bioinformatics and experimental tools and hence represent the first step to understanding NAD+ metabolism in these parasites.

Functional complementation of Leishmania (Leishmania) amazonensis AP endonuclease gene (lamap) in Escherichia coli mutant strains challenged with DNA damage agents

During its life cycle Leishmania spp. face several stress conditions that can cause DNA damages. Base Excision Repair plays an important role in DNA maintenance and it is one of the most conserved mechanisms in all living organisms. DNA repair in trypanosomatids has been reported only for Old World Leishmania species. Here the AP endonuclease from Leishmania (L.) amazonensis was cloned, expressed in Escherichia coli mutants defective on the DNA repair machinery, that were submitted to different stress conditions, showing ability to survive in comparison to the triple null mutant parental strain BW535. Phylogenetic and multiple sequence analyses also confirmed that LAMAP belongs to the AP endonuclease class of proteins.

Biologia de Neotrioza tavaresi Crawford, 1925 (Hemiptera, Psyllidae), galhador da folha do araçazeiro (Psidium cattleianum)

Biology of the leaf gall inducer Neotrioza tavaresi Crawford, 1925 (Hemiptera, Psyllidae) on strawberry guava tree (Psidium cattleianum). A field study was conducted in Curitiba region, State of Paraná, southern Brazil, to describe the life cycle of Neotrioza tavaresi Crawford, 1925, a leaf galling insect in strawberry guava trees (Psidium cattleianum). Three cycles were observed (1997, 1998, 1999) during regular field trips and the insects were observed in Piraquara municipality, where 15 samples with 50 infested leaves were sampled in the 1997-98 cycle. Galls were dissected for detailed studies. Neotrioza tavaresi has a univoltine cycle in which adult individuals were found inside the galls from August onwards. The sexually mature insects with sex ratio 1, emerged from the galls after their dehiscence caused by feeding of the adult insects on the gall walls. Adult emergence started in early October and ended by early December, with its peak in November. Copulation took place as soon as adults exit the gall and egg laying started the next day. Females had more than 100 ovarioles containing 218.7±44.7 (n=50) fully formed eggs. This indicated the short sexual adult life-span (aprox. 5-7 days) of the species, also characterized by a concentrated oviposition. Adult individuals fed and laid their eggs on younger shoots of the plant. The bottoms of the yellowish eggs were inserted into the leaf tissue, mainly on its adaxial edge (78.1%). The nymphs hatched and, as they fed on the adaxial side of expanding leaves, modified the cell growth pattern and the round-shape galls developed on the adaxial side with one insect inside. The gall wall showed distinct layers, with the inner one suppliyng the food to the insects, and the outer layer supplying gall protection. Nymphs went through five instars and the exuviae remained stuck on a ball of wax inside the gall. All parasitoids found were Hymenoptera belonging to Chalcidoidea: Eulophidae (1 sp), Pteromalidae (2 spp) and Encyrtidae (3 spp). The findings suggest that leaf gall inducer and parasitoids insects and plant life cycles are closely connected and both leaf sprouting and gall opening seem to be triggered by the same environmental and plant conditions. The high abundance of shoots may favor insect performance as adult individuals can easily find an ideal place for feeding, copulating and laying eggs.

Aspectos biológicos de Zabrotes subfasciatus (Bohemann, 1833) (Coleoptera, Bruchidae) em Phaseolus vulgaris L., cv. Carioca (Fabaceae), sob condições de laboratório

Zabrotes subfasciatus is a serious pest of common beans, P. vulgaris L.. In Brazil there are several studies dealing with resistance of bean genotypes to this insect, while other studies have emphasized the utilization of oils and powders from plants to repel their attack. In this paper, fecundity, fertility, pattern of oviposition, life cycle and longevity were evaluated for a Brazilian stock from the Goiás State on P. vulgaris cv. Carioca, at 30ºC and 70% R.H. The mean fecundity was 38 eggs per female and 73% of viability. Egg laying showed an aggregated pattern. Males and females lived an average of 13 and 9 days, respectively. The total life cycle lasted for about 28 days.