Diagnóstico molecular de arboviroses brasileiras em soros de pacientes com doença febril aguda

As arboviroses freqüentemente são associadas com surtos nos seres humanos e representam um sério problema de saúde pública, com impactos econômico e social. Entre os arbovírus brasileiros, os vírus Mayaro (MAYV), Dengue (DENV), Febre Amarela (Yellow Fever – YFV), Rocio (ROCV) e Oropouche (OROV) são responsáveis por mais de 95% dos casos de arboviroses humanas no Brasil, causando muitas vezes a mesma sintomatologia clínica, especialmente na fase aguda das infecções. Este estudo teve como objetivo analisar a presença de arbovírus em soros de pacientes com doença febril aguda, durante epidemia de Dengue no município de São José do Rio Preto (SP), bem como de pacientes com doença febril aguda, negativos para Malária e arbovírus do município de Acrelândia (AC). Foi utilizada a técnica de RT-Nested-PCR para a detecção e identificação dos principais arbovírus brasileiros pertencentes aos gêneros Flavivirus, Alphavirus e Orthobunyavirus. Na análise molecular das amostras de São José do Rio Preto, 320 (98%) foram positivas para DENV-3, 3 (1%) para DENV-2, 6 (2%) para o SLEV. Uma destas amostras apresentou co-infecção por DENV-2 e DENV-3 e três amostras apresentaram co-infecção por DENV-3 e SLEV. A alta positividade para o DENV-3 está relacionada com a introdução deste sorotipo no município. Em amostras de Acrelândia, 5 positivas para DENV-3, 1 para MAYV e 2 positivas para OROV. A análise filogenética do gene do envelope mostrou que a amostra isolada de SLEV agrupou-se próxima às estirpes de Belém, que pertencem ao genótipo V. A amostra de OROV agrupou-se ao genótipo I, sendo este o mais distribuído na região Amazônica. Estes dados indicam a circulação de diferentes arbovírus em duas regiões distintas do Brasil e demonstram a necessidade de um sistema de vigilância eficiente para controlar a disseminação destes na população.Arboviruses are frequently associated with outbreaks in humans and represent a serious public health problem, with economic and social impacts. Among the Brazilian arboviruses, Mayaro virus (MAYV), Dengue virus (DENV), Yellow Fever virus (YVF), Rocio virus (ROCV) and Oropouche virus (OROV) are responsible for more than 95% of human cases in Brazil. These arboviruses usually present the same clinical symptoms, especially on the acute phase of infection. The aim of this study was to investigate the presence of arboviruses in serum of patient presenting acute febrile illness, during a dengue outbreak in the city of São José do Rio Preto (state of São Paulo) and the serum of patients who were tested negatively for malaria and arboviruses in the city of Acrelândia (state of Acre). The RT-Nested-PCR technique was applied to detect and to identify the main brazilian arboviruses, especially in Flavivirus, Alphavirus e Orthobunyavirus genera. The molecular analysis in the samples of São José do Rio Preto samples showed that 320 (98%) were positive for DENV-3, 3 (1%) to DENV-2, 6 (2%) to SLEV. Among the positive samples, one presented coinfection with DENV-2 and DENV-3 and three presented coinfection with DENV-3 and SLEV. The high positivity of DENV-3 is associated with the introduction of a new DENV serotype in the city. In the samples from Acrelândia, 5 were positive to DENV-3, 1 to MAYV and 2 positive to OROV. The phylogenetic analysis of the envelope gene from a positive SLEV sample indicated that the virus circulating in São José do Rio Preto is most related to strains from Belém, which belong to lineage V. The OROV belongs to lineage I, the most widespread in Amazon region. These results indicate the circulation of different arboviruses in two distinct regions of Brazil and they demonstrate the need of an efficient surveillance system to control the dissemination of these arbovirus in the population

Avaliação da caspase-3 e Ki-67 como marcadores prognósticos nas neoplasias mamárias em cadelas

Apoptosis, as a cellular event, has an important participation in tumorigenesis, determining the tumoral growth and aggressiveness. The present study had as objective to evaluate the occurrence of apoptosis, associated cellular proliferation, in canine mammary neoplasias and the patient clinical evolution. Seventy dogs had been submitted to surgical excision of the tumor fragment and they were submitted to the histopathologic diagnosis and imunohistochemistry procedure to caspase-3 and the Ki-67. This apoptosis and cellular proliferation markers demonstrated great expression in malignant neoplasias especially carcinoma, considered the most malignant of the tumors. These results confirm consulted literature contributing for a criterious tumoral prognostic complementing the tumoral classification, providing a greater and better supervened due to adequacy of therapeutic procedure of each patient.A apoptose e a proliferação celular possuem uma participação importante na tumorigênese, determinando o crescimento tumoral e consequentemente sua agressividade. O presente estudo teve como objetivo avaliar a ocorrência da apoptose associada a proliferação celular em neoplasias mamárias em cadelas e a elas a evolução clínica do paciente. Setenta animais foram submetidos à exérese cirúrgica do tumor, sendo este submetido ao diagnóstico histopatológico e marcação imuno-histoquímica para caspase-3 e Ki-67. Estes marcadores de apoptose e proliferação celular demonstraram grande expressão nas neoplasias malignas, principalmente nos carcinomas, considerado o mais maligno dos tumores. Estes resultados corroboram os dados da literatura e contribuem para um prognóstico tumoral criterioso que complementa a classificação tumoral existente proporcionando uma melhor e maior sobrevida devido a uma adequação do procedimento terapêutico de cada paciente

Concurrent dengue and malaria in the Amazon region

Introduction: The Amazon region has extensive forested areas and natural ecosystems, providing favorable conditions for the existence of innumerous arboviruses. Over 200 arboviruses have been isolated in Brazil and about 40 are associated with human disease. Four out of 40 are considered to be of public health importance in Brazil: Dengue viruses (1-4), Oropouche, Mayaro and Yellow Fever. Along with these viruses, about 98% of the malaria cases are restricted to the Legal Amazon region. Methods: This study aimed to investigate the presence of arboviruses in 111 clinical serum samples from patients living in Novo Repartimento (Para), Placido de Castro (Acre), Porto Velho (Rondonia) and Oiapoque (Amapa). The viral RNA was extracted and RT-PCR was performed followed by a Multiplex-Nested-PCR, using Flavivirus, Alphavirus and Orthobunyavirus generic and species-specific primers. Results: Dengue virus serotype 2 was detected in two patients living in Novo Repartimento (Para) that also presented active Plasmodium vivax infection. Conclusions: Despite scant data, this situation is likely to occur more frequently than detected in the Amazon region. Finally, it is important to remember that both diseases have similar clinical findings, thus the diagnosis could be made concomitantly for dengue and malaria in patients living or returning from areas where both diseases are endemic or during dengue outbreaks.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

First genome sequence of St. Louis encephalitis virus (SLEV) isolated from a human in Brazil

St. Louis encephalitis virus (SLEV), a member of the family Flaviviridae, genus Flavivirus, is a causative agent of encephalitis in the Americas. In Brazil, sporadic cases of SLEV infection have been reported since 1953, but the first outbreak of SLEV in Brazil was identified only in 2007, concomitant with an outbreak of dengue virus (DENV) serotype 3. This finding, along with other reports, indicates that SLEV circulation in Brazil is largely unknown, and there may be epidemiological implications of the co-circulation of SLEV, DENV and other flaviviruses in Brazil. Here, we describe the first complete genome sequence of an SLEV strain isolated from a human patient in Brazil, strain BeH 355964. Phylogenetic analysis was performed to determine the genotype of BeH 355964 using the full-length genome and envelope (E) gene sequences separately. Both analyses showed that BeH 355964 could be classified as genotype V. Although the number of single gene sequences available is greater (such as for the E gene), the phylogenetic tree based on the complete genome sequence was better supported and provided further information about the virus.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Isolation and characterization of mayaro virus from a human in Acre, Brazil

Mayaro virus (MAYV) is widely distributed throughout South America and is the etiologic agent of Mayaro fever, an acute febrile illness often presenting with arthralgic manifestations. The true incidence of MAYV infection is likely grossly underestimated because the symptomatic presentation is very similar to that of dengue fever and other acute febrile tropical diseases. We report the complete genome sequence of a MAYV isolate detected from an Acrelandia patient presenting with fever, chills, and sweating, but with no arthralgia. Results show that this isolate belongs to genotype D and is closely related to Bolivian strains. Our results suggest that the Acre/Mayaro strain is closely related to the progenitor of these Bolivian strains that were isolated between 2002 and 2006.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP