A novel trypsin Kazal-type inhibitor from Aedes aegypti with thrombin coagulant inhibitory activity

Kazal-type inhibitors play several important roles in invertebrates, such as anticoagulant, vasodilator and antimicrobial activities. Putative Kazal-type inhibitors were described in several insect transcriptomes. In this paper we characterized for the first time a Kazal unique domain trypsin inhibitor from the Aedes aegypti mosquito. Previously, analyses of sialotranscriptome of A. aegypti showed the potential presence of a Kazal-type serine protease inhibitor, in female salivary glands, carcass and also in whole male, which we named AaTI (A. aegypti trypsin inhibitor). AaTI sequence showed amino acid sequence similarity with insect thrombin inhibitors, serine protease inhibitor from Litopenaeus vannamei hemocytes and tryptase inhibitor from leech Hirudo medicinalis (LDTI). In this work we expressed, purified and characterized the recombinant AaTI (rAaTI). Molecular weight of purified rAaTI was 7 kDa rAaTI presented dissociation constant (K(i)) of 0.15 and 3.8 nM toward trypsin and plasmin, respectively, and it weakly inhibited thrombin amidolytic activity. The rAaTI was also able to prolong prothrombin time, activated partial thromboplastin time and thrombin time. AaTI transcription was confirmed in A. aegypti female salivary gland and gut 3 h and 24 h after blood feeding, suggesting that this molecule can act as anticoagulant during the feeding and digestive processes. Its transcription in larvae and pupae suggested that AaTI may also play other functions during the mosquito`s development. (C) 2010 Elsevier Masson SAS. All rights reserved.

Estrutura populacional do camarão-branco Litopenaeus schmitti nas regiões estuarina e marinha da baixada santista, São Paulo, Brasil

The white-shrimp Litopenaeus schmitti distributes in West Atlantic Ocean, occurring along all Brazilian cost. Population structure in the Baixada Santista region was identified from samples obtained from artisanal and industrial fishery between June of 2005 and May of 2006. A total of 2.912 specimens were collected, being 2.138 females (1.008 in the estuary and 1.130 in the marine region) and 774 males ( 334 in the estuary and 440 in the marine region). Environmental parameters were annotated together the sampling, allowing to identify that water temperature influences directly the catches. Catches variations, length composition of samples by sex and gonadal maturation of females allowed to identify that: (i) estuary is used as a nursery area by individuals with small lengths, most young; (ii) marine region is used by larger individuals ( adults) and the spawning period extend from June to February, mainly between November and January. It was verified that estuarine fishery ( artisanal) focuses immature and in development individuals, with small lengths and, the marine fishery ( industrial) focuses adults during the whole year and, only in the summer, youngling from spawn. The length of first gonadal maturation of females was estimated in 15,8mm. These results and diagnoses must be considered in the management of L. schmitti fishery in Baixada Santista region.

Cytotoxic activity of a dichloromethane extract and fractions obtained from Eudistoma vannamei (Tunicata: Ascidiacea)

This study consists of the bioassay-guided fractionation of the dichloromethane extract from Eudistoma vannamei and the pharmacological characterization of the active fractions. The dried hydromethanolic extract dissolved in aqueous methanol was partitioned with dichloromethane and chromatographed on a silica gel flash column. The anti-proliferative effect was monitored by the MTT assay. Four of the latest fractions, numbered 14 to 17, which held many chemical similarities amongst each other, were found to be the most active. The selected fractions were tested for viability, proliferation and death induction on cultures of HL-60 promycloblastic leukemia cells. The results suggested that the observed cytotoxicity is related to apoptosis induction. (C) 2007 Elsevier Inc. All rights reserved.