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1.
J Virol Methods ; 225: 4-8, 2015 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-26342906

RESUMO

Porcine epidemic diarrhea virus (PEDV) causes acute diarrhea and dehydration in sucking piglets and has a high mortality rate. An immunochromatography (IC) assay, known as a lateral flow test, is a simple device intended to detect the presence of target pathogens. Here, we developed an IC assay that detected PEDV antigens with 96.0% (218/227) sensitivity and 98.5% (262/266) specificity when compared with real-time reverse transcriptase (RT)-PCR using FAM-labeled probes based on sequences from nucleocapsid genes. The detection limits of the real-time RT-PCR and IC assays were 1×10(2) and 1×10(3) copies, respectively. The IC assay developed herein did not detect non-specific reactions with other viral or bacterial pathogens, and the assay could be stored at 4°C or room temperature for 15 months without affecting its efficacy. Thus, the IC assay may result in improved PED detection and control on farms, and is a viable alternative to current diagnostic tools for PEDV.


Assuntos
Cromatografia de Afinidade/métodos , Infecções por Coronavirus/veterinária , Vírus da Diarreia Epidêmica Suína/isolamento & purificação , Doenças dos Suínos/diagnóstico , Doenças dos Suínos/virologia , Animais , Antígenos Virais/análise , Infecções por Coronavirus/diagnóstico , Infecções por Coronavirus/virologia , Reação em Cadeia da Polimerase em Tempo Real , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sensibilidade e Especificidade , Suínos
2.
Arch Pharm Res ; 32(10): 1469-73, 2009 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-19898812

RESUMO

The aim of this study was to provide a basis for examining the molecular mechanism for the pharmacological action of ethanol. Energy transfer between the surface fluorescent probe 1-anilinonaphthalene-8-sulfonic acid and hydrophobic fluorescent probe 1,3-di(1-pyrenyl)propane was used to examine the effect of both dimyristoylphosphatidylethanol (DMPEt) and ethanol on the thickness (D) of the synaptosomal plasma membrane vesicles (SPMV) isolated from the bovine cerebral cortex. The thickness (D) of the intact SPMV was 1.044 +/- 0.008 (arbitrary units, n=5) at 37 degrees C (pH 7.4). Both DMPEt and ethanol decreased the thickness of the SPMV lipid bilayer in a dose-dependent manner with a significant decrease in thickness observed at 5 microM and 25 mM, respectively. It was assumed that both ethanol and DMPEt cause interdigitation in the SPMV lipid bilayers. The effects of ethanol on the neuronal membranes were attributed to its direct and indirect actions. The indirect action of ethanol refers to the action of phosphatidylethanol, which is an ethanol abnormal metabolite, on the neuronal membranes. The decrease in membrane thickness by both DMPEt and ethanol might be responsible for some, but not all of its anesthetic actions.


Assuntos
Membrana Celular/efeitos dos fármacos , Etanol/farmacologia , Glicerofosfolipídeos/farmacologia , Bicamadas Lipídicas/metabolismo , Neurônios/efeitos dos fármacos , Sinaptossomos/efeitos dos fármacos , Animais , Bovinos , Membrana Celular/metabolismo , Membrana Celular/ultraestrutura , Córtex Cerebral/citologia , Córtex Cerebral/efeitos dos fármacos , Córtex Cerebral/metabolismo , Relação Dose-Resposta a Droga , Transferência de Energia/efeitos dos fármacos , Técnicas In Vitro , Neurônios/metabolismo , Neurônios/ultraestrutura , Sinaptossomos/metabolismo , Sinaptossomos/ultraestrutura
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