RESUMEN
We report here the whole-genome sequence of the first camelpox virus case diagnosed in Israel. The strain (Negev2016) was isolated in 2016 from a camel in southern Israel and was sequenced on the Illumina MiSeq and Oxford Nanopore MinION platforms.
RESUMEN
An outbreak of a disease in camels with skin lesions was reported in Israel during 2016. To identify the etiological agent of this illness, we employed a multidisciplinary diagnostic approach. Transmission electron microscopy (TEM) analysis of lesion material revealed the presence of an orthopox-like virus, based on its characteristic brick shape. The virus from the skin lesions successfully infected chorioallantoic membranes and induced cytopathic effect in Vero cells, which were subsequently positively stained by an orthopox-specific antibody. The definite identification of the virus was accomplished by two independent qPCR, one of which was developed in this study, followed by sequencing of several regions of the viral genome. The qPCR and sequencing results confirmed the presence of camelpox virus (CMLV), and indicated that it is different from the previously annotated CMLV sequence available from GenBank. This is the first reported case of CMLV in Israel, and the first description of the isolated CMLV subtype.
Asunto(s)
Orthopoxvirus , Infecciones por Poxviridae/epidemiología , Infecciones por Poxviridae/virología , Animales , Línea Celular , Embrión de Pollo , Chlorocebus aethiops , Efecto Citopatogénico Viral , Brotes de Enfermedades , Femenino , Humanos , Israel/epidemiología , Orthopoxvirus/clasificación , Orthopoxvirus/genética , Orthopoxvirus/ultraestructura , Filogenia , Vigilancia de la Población , Infecciones por Poxviridae/diagnóstico , Células VeroRESUMEN
Gonadotropin releasing hormone-I (GnRH-I), a decapeptide serves as a key regulator of reproduction. Recently, several groups have identified in the mammalian brain a second form of GnRH, of unknown function, designated GnRH-II. The human neuronal medulloblastoma cells (TE-671) were recently demonstrated to express the two forms of GnRH (GnRH-I and GnRH-II). We used this cell line, as a model system, to investigate the regulation of human GnRH-I and GnRH-II genes by estrogen. Estrogen is one of the principal regulators of GnRH-I in hypothalamic neurons, acting as a classic homeostatic feedback molecule between the gonads and the brain. In this study, we investigated the regulation of the two GnRH forms by estrogen, in the human neuronal cell line TE-671. We demonstrate, for the first time, that the hGnRH-II and hGnRH-I genes are differentially regulated by estrogen. Using reverse transcriptase-polymerase chain reaction (RT-PCR) and Southern hybridization, we found that estrogen increases endogenous hGnRH-II mRNA levels and decreases endogenous hGnRH-I mRNA levels. Furthermore, we found these effects to be promoter-mediated. We cloned the hGnRH-I and hGnRH-II promoter constructs upstream to a luciferase reporter plasmid, and cotransfected these constructs with an estrogen receptor alpha into the TE-671 neuronal cells. Luciferase activity of GnRH promoter constructs treated with estrogen demonstrates that the differential regulation of the GnRH genes by estrogen is mediated at the transcription level.