No abstract

Dengue serotype 3 viruses were isolated from patients in Brazil from 2002 through 2004. On the basis of phylogenetic analyses, these isolates were assigned genotype 1. This genotype had never been reported in South America before. Its appearance indicates a major risk factor for dengue epidemics and severe disease.

The lack of an accurate diagnosis has been a serious obstacle to the advancement of the anti-Trypanosoma cruzi chemotherapy and long-term infection can result in different health risks to human. PCRs are alternative methods, more sensitive than conventional parasitological techniques, which due to their low sensitivities are considered unsuitable for these purposes. The aim of this study was to investigate a sensitive diagnostic strategy to quantify blood and cardiac tissues parasites based on real-time PCR tools during acute and chronic phases of murine Chagas disease, as well as to monitor the evolution of infection in those mice under specific treatment. In parallel, fresh blood examination, immunological analysis and quantification of cardiac inflammation were also performed to confront and improve real-time PCR data. Similar profiles of parasitemia curves were observed in both quantification techniques during the acute phase of the infection. In contrast, parasites could be quantified only by real-time PCR at 60 and 120 days of infection. In cardiac tissue, real-time PCR detected T. cruzi DNA in 100% of infected mice, and using this tool a significant Pearson correlation between parasite load in peripheral blood and in cardiac tissue during acute and chronic phases was observed. Levels of serum CCL2, CCL5 and nitric oxide were coincident with parasite load but focal and diffuse mononuclear infiltrates was observed, even with significant (p<0.05) reduction of parasitism after 60 days of infection. Later, this methodology was used to monitor the evolution of infection in animals treated with itraconazole (Itz). Itz-treatment induced a reduction of parasite load in both blood and cardiac muscle at the treatment period, but after the end of chemotherapy an increase of parasitism was detected. Interestingly, inflammatory mediators levels and heart inflammation intensity had similar evolution to the parasite load, in the group of animals treated. Taken together, our data show that real-time PCR strategy used was suitable for studies of murine T. cruzi infection and may prove useful in investigations involving experimental chemotherapy of the disease and the benefits of treatment in relation to parasitism and inflammatory response.

No abstract

The mosquito Stegomyia albopicta is among the most important arbovirus vectors in the world, particularly for Dengue viruses. Their natural history suggests that biologically these viruses are highly adapted to their mosquito hosts and they were most likely mosquito viruses prior to becoming adapted to lower primates and humans. As well as being maintained by transmission among susceptible humans, Dengue viruses may also be maintained by vertical transmission in mosquitoes during inter-epidemic periods. The larvae and mosquitoes of Stegomyia albopicta were used to identify the vertical transmission of the dengue virus in nature and to confirm the vectorial capacity concerning the Dengue virus type 2 infection. The minimum infection rate concerning S. albopicta infection with the Dengue virus was 1:36.45. In Brazil this was the first time that high minimum infection rates of vertical transmission of S. albopicta were detected in this species.Keywords: Dengue virus, vertical transmission, Stegomyia albopicta, vectorial capacity. Transmissão vertical natural de Stegomyia albopicta como vetor de dengue no BrasilResumo O mosquito Stegomyia albopicta está dentre os mais importantes vetores de arbovírus do mundo, particularmente para o Dengue virus. A história natural sugere que biologicamente esses vírus são altamente adaptados aos seus mosquitos vetores e foram, provavelmente, os vírus que infectavam mosquitos antes de se tornarem adaptados a primatas não humanos e humanos. Além de serem mantidos entre os homens susceptíveis, os Dengue viruses podem também ser mantidos pela transmissão vertical em mosquitos durante os períodos interepidêmicos. As larvas e mosquitos da espécie Stegomyia albopicta foram utilizados para a identificação da transmissão vertical do vírus dengue na natureza e para confirmar a capacidade vetorial em relação à infecção pelo DENV-2 infection. A taxa mínima de infecção em relação à infecção do S. albopicta com o Dengue virus foi de 1:36,45. No Brasil, esta é a primeira vez que altas taxas de infecção mínima da transmissão vertical de S. albopicta foram detectadas nessa espécie.Palavras-chave: Dengue vírus, transmissão vertical, Stegomyia albopicta, capacidade vetorial.

We report the detailed molecular characterization of two PLA2s, Lys49 and Asp49 isolated from Bothrops leucurus venom, and examined their effects against Dengue virus (DENV). The Bl-PLA2s, named BlK-PLA2 and BlD-PLA2, are composed of 121 and 122 amino acids determined by automated sequencing of the native proteins and peptides produced by digestion with trypsin. They contain fourteen cysteines with pIs of 9.05 and 8.18 for BlK- and BlD-PLA2s, and show a high degree of sequence similarity to homologous snake venom PLA2s, but may display different biological effects. Molecular masses of 13,689.220 (Lys49) and 13,978.386 (Asp49) were determined by mass spectrometry. DENV causes a prevalent arboviral disease in humans, and no clinically approved antiviral therapy is currently available to treat DENV infections. The maximum non-toxic concentration of the proteins to LLC-MK2 cells determined by MTT assay was 40 µg/mL for Bl-PLA2s (pool) and 20 µg/mL for each isoform. Antiviral effects of Bl-PLA2s were assessed by quantitative Real-Time PCR. Bl-PLA2s were able to reduce DENV-1, DENV-2, and DENV-3 serotypes in LLC-MK2 cells infection. Our data provide further insight into the structural properties and their antiviral activity against DENV, opening up possibilities for biotechnological applications of these Bl-PLA2s as tools of research.

No abstract

No abstract