ABSTRACT
BACKGROUND Gene reporter-fluorescent cells have emerged as alternative method for drug screening. OBJECTIVE Achievement of constitutive expression of fluorescent protein GFP by Leishmania braziliensis as alternative method for drug screening. METHODS L. braziliensis-GFP was generated using Leishmania tarentolae pLEXSY-egfp for constitutive expression of GFP. Fluorescent cells were selected and subjected to standardisation tests of anti-promastigote and anti-intracellular amastigote assays. FINDINGS Our results showed that L. braziliensis-GFP method is faster and more sensitive than Allamar Blue-resazurin. MAIN CONCLUSION Transfected parasites maintained stable fluorescence after successive in vitro passages and pLEXSY system can be used to achieve non-L. tarentolae fluorescent cells.
Subject(s)
Animals , Leishmania braziliensis/drug effects , Amphotericin B/pharmacology , Green Fluorescent Proteins/pharmacology , Drug Evaluation, Preclinical/methods , Antiprotozoal Agents/pharmacology , Time Factors , Microscopy, FluorescenceABSTRACT
Porcine circovirus-2 (PCV-2) infection is currently considered an important disease of swine. The pathogenic agent was first described in Brazil in 2000. This study detected the PCV-2 DNA in four Brazilian pig tissues collected between 1978 and 1979. This observation is the oldest description of this virus in Brazil.
Subject(s)
Animals , Circoviridae Infections , Circovirus , Diagnostic Techniques and Procedures , Polymerase Chain Reaction , Swine , Methods , MethodsABSTRACT
Este trabalho mostrou a purificação de uma apirase em extratos de tegumentos de vermes adultos de S. mansoni com razão de hidrólise de ADP/ATP aproximadamente 2. Análise da amostra por MALDI-TOF evidenciou a presença de helicase e carboxilesterase, sendo que a atividade de hidrólise de ADP ainda não foi descrita para estas proteínas. Através de ®Western blot¼ mostrou-se que a proteína purificada não tem epítopos reconhecíveis por anticorpo anti-apirase de batata, e que a imunoreatividade cruzada com apirase da família CD39 está presente na fração insolúvel que não foi utilizada no processo de purificação. Esta tese caracteriza também a utilização de ®primers¼ consenso degenerados na construção de bibliotecas de cDNA com PCR de baixa estringência...