Triagem molecular de portadores assintomáticos de Plasmodium sp. entre Bancos de Sangue da região Amazônica brasileira

The transmission of malaria in Brazil is heterogeneous throughout endemic areas and the presence of asymptomatic Plasmodium sp. carriers (APCs) in the Brazilian Amazon has already been demonstrated. Malaria screening in blood banks is based on the selection of donors in respect to possible risks associated with travel or residence, clinical evidence and/or inaccurate diagnostic methods thereby increasing the probability of transfusion-transmitted infection. We evaluated the frequency of APCs in four blood services in distinct areas of the Brazilian Amazon region. DNA was obtained from 400 human blood samples for testing using the phenol-chloroform method followed by a nested-PCR protocol with species-specific primers. The positivity rate varied from 1 to 3% of blood donors from the four areas with an average of 2.3%. All positive individuals had mixed infections for Plasmodium vivax and Plasmodium falciparum. No significant differences in the results were detected among these areas; the majority of cases originated from the transfusion centres of Porto Velho, Rondônia State and Macapá, Amapá State. Although it is still unclear whether APC individuals may act as reservoirs of the parasite, efficient screening of APCs and malaria patients in Brazilian blood services from endemic areas needs to be improved.A transmissão da malária no Brasil é heterogênea em todas as áreas endêmicas e a presença de portadores assintomáticos de Plasmodium sp. (PAPs) na Amazônia brasileira já foi demonstrada. A triagem de pacientes maláricos em bancos de sangue é baseada na seleção dos doadores com relação aos riscos possíveis associados com residência, evidência clínica e/ou os métodos diagnósticos não acurados que aumentam a probabilidade da infecção transmitida por transfusão. Avaliamos a freqüência de PAPs em quatro bancos de sangue em áreas distintas da região Amazônica brasileira. O DNA foi obtido a partir de 400 amostras de sangue humano usando o método do fenol-clorofórmio, seguido por um protocolo de nested-PCR com oligonucleotídeos espécie-específicos. A taxa de positividade variou de 1 a 3% de doadores do sangue das quatro áreas, com uma média de 2,3%. Todos os indivíduos positivos tinham infecções mistas entre o Plasmodium vivax e o Plasmodium falciparum. Nenhuma diferença significativa nos resultados foi detectada entre estas áreas; a maioria dos casos originou dos Hemocentros de Porto Velho, do Estado de Rondônia e de Macapá, Estado do Amapá. Embora ainda não esteja claro se os indivíduos PAPs possam agir como reservatórios do parasito, a triagem eficiente de PAPs e de pacientes com malária em bancos de sangue no Brasil das áreas endêmicas necessita ser implementada

Angiogenic properties of dental pulp stem cells conditioned medium on endothelial cells in vitro and in rodent orthotopic dental pulp regeneration

Objectives: To evaluate the effect of SHED-CM on the proliferation, differentiation, migration ability, cell death, gene expression and production of VEGF of HUVEC in vitro and in a rodent orthotopic dental pulp regeneration. Methods: Three culture media [M199, DMEM/Ham's F12 and DMEM/Ham's F12 conditioned by SHEDs] were used as experimental groups. SHED-CM was prepared maintaining confluent cells in culture without serum for 3 days. The proliferation and cell death marker of HUVECs were assessed using flow cytometry. The capacity of formation of vascular-like structures was analyzed in cells grown over Matrigel® in hypoxic condition. HUVECs migration was followed using the scratch test. VEGF-A expression in HUVECs was assessed using real time RT-qPCR; and VEGF synthesis with ELISA test. SHED-CM was also applied in rodent ortotopic model of dental pulp regeneration in rats. The formed tissue was submitted to histological and immunohistochemical analyses. Results: SHED-CM promoted significantly lower expression of 7AAD in HUVECs; whereas the expression of the Ki67 was similar in all groups. The vascular-like structures were observed in all groups. Migration of SHED-CM group was faster than DMEM/Ham's F12. SHED-CM induced similar expression of VEGF-A than M199, and higher than DMEM/Ham's F12. SHED-CM induced significantly higher VEGF synthesis than other media. SHED-CM induced formation of a vascularized connective tissue inside the root canal. Conclusion: The study showed that SHEDs release angiogenic and cytoprotective factors, which are of great importance for tissue engineering. Clinical significance: SHED-CM could be an option to the use of stem cells in tissue engineering

Molecular screening of Plasmodium sp. asymptomatic carriers among transfusion centers from Brazilian Amazon region

A transmissão da malária no Brasil é heterogênea em todas as áreas endêmicas e a presença de portadores assintomáticos de Plasmodium sp. (PAPs) na Amazônia brasileira já foi demonstrada. A triagem de pacientes maláricos em bancos de sangue é baseada na seleção dos doadores com relação aos riscos possíveis associados com residência, evidência clínica e/ou os métodos diagnósticos não acurados que aumentam a probabilidade da infecção transmitida por transfusão. Avaliamos a freqüência de PAPs em quatro bancos de sangue em áreas distintas da região Amazônica brasileira. O DNA foi obtido a partir de 400 amostras de sangue humano usando o método do fenol-clorofórmio, seguido por um protocolo de nested-PCR com oligonucleotídeos espécie-específicos. A taxa de positividade variou de 1 a 3% de doadores do sangue das quatro áreas, com uma média de 2,3%. Todos os indivíduos positivos tinham infecções mistas entre o Plasmodium vivax e o Plasmodium falciparum. Nenhuma diferença significativa nos resultados foi detectada entre estas áreas; a maioria dos casos originou dos Hemocentros de Porto Velho, do Estado de Rondônia e de Macapá, Estado do Amapá. Embora ainda não esteja claro se os indivíduos PAPs possam agir como reservatórios do parasito, a triagem eficiente de PAPs e de pacientes com malária em bancos de sangue no Brasil das áreas endêmicas necessita ser implementada.The transmission of malaria in Brazil is heterogeneous throughout endemic areas and the presence of asymptomatic Plasmodium sp. carriers (APCs) in the Brazilian Amazon has already been demonstrated. Malaria screening in blood banks is based on the selection of donors in respect to possible risks associated with travel or residence, clinical evidence and/or inaccurate diagnostic methods thereby increasing the probability of transfusion-transmitted infection. We evaluated the frequency of APCs in four blood services in distinct areas of the Brazilian Amazon region. DNA was obtained from 400 human blood samples for testing using the phenol-chloroform method followed by a nested-PCR protocol with species-specific primers. The positivity rate varied from 1 to 3% of blood donors from the four areas with an average of 2.3%. All positive individuals had mixed infections for Plasmodium vivax and Plasmodium falciparum. No significant differences in the results were detected among these areas; the majority of cases originated from the transfusion centres of Porto Velho, Rondônia State and Macapá, Amapá State. Although it is still unclear whether APC individuals may act as reservoirs of the parasite, efficient screening of APCs and malaria patients in Brazilian blood services from endemic areas needs to be improved.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq