RESUMO
Zika virus (ZIKV) has emerged globally as an important pathogen, since it has been recognized as a cause of microcephaly and other neurologic processes and sequalae in newborns. The virus shares homology with Hepaciviruses and therefore may be a cause of hepatitis. We sought to characterize ZIKV replication in hepatocyte-derived cell lines. Huh7.5 and HepG2 cells were infected with ZIKV and replication potential was evaluated by multiple methods including plaque assay, qRT-PCR, negative-strand ZIKV RNA production, and ZIKV NS1 protein production. Growth curves in cells and supernatant were compared to replicative capacity in Vero cells. Overall, viral replication in both hepatocyte lines approximated that observed in the Vero cells. Cell cytopathology was observed after 3 days of infection and apoptosis markers increased. Transmission electron microscopy revealed evidence of viral capsids in cells and negative staining revealed ZIKV particles in the supernatant. Conclusions: Hepatocyte-derived cell lines are permissive for ZIKV replication and produce an overt cytopathic effect consistent with development of an acute viral hepatitis. Further evaluation of replication and injury is warranted.
Assuntos
Fígado/virologia , Infecção por Zika virus/virologia , Zika virus/genética , Animais , Linhagem Celular , Linhagem Celular Tumoral , Chlorocebus aethiops , Efeito Citopatogênico Viral/genética , Células Hep G2 , Hepatócitos/virologia , Humanos , Células Vero , Carga Viral/genética , Proteínas não Estruturais Virais/genética , Vírion/genética , Replicação Viral/genéticaAssuntos
Infecções por HIV/virologia , Zika virus/imunologia , Adulto , Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/sangue , Estudos de Coortes , Vírus da Dengue/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Gana/epidemiologia , Infecções por HIV/epidemiologia , Infecções por HIV/imunologia , Humanos , Masculino , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Zika virus/genéticaRESUMO
The field of centrifugal microfluidics has experienced tremendous growth during the past 15 years, especially in applications such as lab-on-a-disc (LoD) diagnostics. The strength of LoD systems lies in its potential for development into fully integrated sample-to-answer analysis systems. This review highlights the technologies necessary to develop the next generation of these systems. In addition to outlining valving and other fluid-handling operations, we discuss the recent advances and future outlook in four categories of LoD processes: reagent storage, sample preparation, nucleic acid amplification, and analyte detection strategies.
Assuntos
Automação Laboratorial/métodos , Microfluídica/métodos , Técnicas de Diagnóstico Molecular/métodos , Microfluídica/tendências , Técnicas de Diagnóstico Molecular/tendências , Manejo de Espécimes/métodos , Manejo de Espécimes/tendênciasRESUMO
This study characterizes a model of motor neuron (MN) loss on the molecular, cellular, and behavioral levels. Injection of the toxic lectin Ricinus communis agglutinin I (RCA I or ricin) caused cellular deficit and loss of function by damaging the sciatic nerve. Since the sciatic nerve supplies movement to most of the lower limb, damaging this motor system models lower limb paralysis and the deficits that occur in diseases like amyotrophic lateral sclerosis (ALS) and infantile progressive spinal muscular atrophy (SMA). We used motor-, sensorimotor-, locomotor-, and reflex-based tests to demonstrate loss of function after ricin injection. Loss of function was also demonstrated by decreased retrograde transport, and supported by measurements of muscle wasting. Histochemical and molecular methods were used to characterize sciatic nerve damage in axons and cell bodies, including apoptotic cell death in MNs. This battery of tests documents the extent of the ricin-induced damage and provides a baseline that can be used to judge the efficacy of MN treatment strategies in preclinical studies.
