RESUMO
The infectious pancreatic necrosis virus (IPNV) is responsible for significant economic losses in the aquaculture industry. It is an unenveloped virus with an icosahedral capsid. Its viral genome comprises two dsRNA segments, A and B. Segment A contains a small ORF, which encodes VP5, and a large ORF, which encodes a polyprotein that generates the structural proteins and the viral protease. Segment B encodes the RNA-dependent RNA polymerase (RdRp), called VP1 in this free form, or Vpg when it covalently attaches to the viral RNA. The viral genome does not have cap or poly(A). Instead, each 5' end is linked to the Vpg. Recently, we demonstrated that mRNA-A contains an internal ribosome entry site (IRES) to command polyprotein synthesis. However, the presence of Vpg on IPNV mRNAs and its impact on cellular translation has not been investigated. This research demonstrates that IPNV mRNAs are linked to Vpg and that this protein inhibits cap-dependent translation on infected cells. Also, it is demonstrated that Vpg interacts with eIF4E and that rapamycin treatment partially diminishes the viral protein synthesis. In addition, we determined that an IRES does not command translation of IPNV mRNA-B. We show that VPg serves as a cap substitute during the initiation of IPNV translation, contributing to understanding the replicative cycle of Birnaviruses. Our results indicate that the viral protein VP1/Vpg is multifunctional, having a significant role during IPNV RNA synthesis as the RdRp and the primer for IPNV RNA synthesis and translation as the viral protein genome, acting as a cap substitute.
Assuntos
Vírus da Necrose Pancreática Infecciosa , Vírus da Necrose Pancreática Infecciosa/genética , Sítios Internos de Entrada Ribossomal , Poliproteínas/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA Viral/genética , RNA Viral/metabolismo , RNA Polimerase Dependente de RNA/genética , Proteínas Virais/genética , Proteínas Virais/metabolismoRESUMO
Due to the COVID-19 pandemic, many transport kits have been manufactured to preserve and transport nasopharyngeal swab samples (NPSs) from patients. However, there is no information on the performance of the different virus transport media (VTM) used in COVID-19 diagnosis in the population of Santiago de Chile. We compared the RT-qPCR amplification profile of five different viral transport kit mediums, including DNA/RNA Shield™, NAT, VTM-N, Ezmedlab™, and phosphate-buffered saline (PBS), for NPSs from Central Metropolitan Health Service, Santiago, Chile. The DNA/RNA Shield™ medium showed a better performance in terms of Cq and RFU values for the internal reference RNase P and viral ORF1ab probes. By contrast, the PBS transport medium registered higher Cq values for the viral and reference gene, compared to the other VTM. DNA/RNA Shield™ shows higher relative fluorescence units (RFUs) and lower Cq values for the reference gene. Collectively, our results suggest that the PBS medium could compromise the sample diagnosis because of its lower RT-qPCR performance. The NAT, Ezmedlab and VTM-N, and DNA/RNA Shield™ media show acceptable RT-qPCR parameters and, consequently, seem suitable for use in COVID-19 diagnosis.
Assuntos
COVID-19 , COVID-19/diagnóstico , Teste para COVID-19 , Chile , Meios de Cultura , Humanos , Nasofaringe , Pandemias , RNA , RNA Viral/análise , RNA Viral/genética , Reação em Cadeia da Polimerase em Tempo Real , SARS-CoV-2/genética , Manejo de Espécimes/métodosRESUMO
Adjuvants used in vaccine aquaculture are frequently harmful for the fish, causing melanosis, granulomas and kidney damage. Along with that, vaccines are mostly administered by injection, causing pain and stress to the fish. We used the DNA coding for the replicase of alphavirus as adjuvant (Ad) of a vaccine against ISAV. The Ad and an inactivated ISAV (V) were loaded in chitosan nanoparticles (NPs) to be administered orally to Atlantic salmon. NP-Ad was able to deliver the DNA ex vivo and in vivo. Oral administration of the NPs stimulated the expression of immune molecules, but did not stimulate the humoral response. Although the vaccination with NP-V results in a modest protection of fish against ISAV, NP-V administered together with NP-Ad caused a protection of 77%. Therefore, the DNA coding for the replicase of alphavirus could be administered orally and can potentiate the immuneprotection of a virine against infection.
Assuntos
Doenças dos Peixes/prevenção & controle , Isavirus/imunologia , Nanopartículas/química , Infecções por Orthomyxoviridae/veterinária , Salmo salar , Vacinas Virais/farmacologia , Adjuvantes Imunológicos/administração & dosagem , Adjuvantes Imunológicos/farmacologia , Administração Oral , Alphavirus/genética , Alphavirus/imunologia , Animais , Doenças dos Peixes/virologia , Infecções por Orthomyxoviridae/prevenção & controle , Infecções por Orthomyxoviridae/virologia , Replicon , Vacinas de Produtos Inativados/imunologiaRESUMO
El envejecimiento de las población ha generado un aumento de adultos mayores, donde gran porcentaje de éstos son mujeres. También se han incrementado las enfermedades crónicas, siendo más frecuentes en este rango etario las patologías articulares, que impactan la funcionalidad y la calidad de vida, contribuyendo a la creencia de mitos en torno a la vejez. No obstante, esta visión cambiaría debido a la creación de una propuesta política gubernamental, la de Envejecimiento Positivo orientada a la inclusión del adulto mayor en la sociedad, mejorando su calidad de vida y posicionándolo como un sujeto de derecho.La Terapia Ocupacional promueve la participación ocupacional de adultas mayores con enfermedades articulares, para que tengan un envejecimiento positivo, aunque para alcanzar ese propósito, es preciso analizar el significado construido que tienen estas personas sobre dicho concepto, por esto se realizó una entrevista semiestructurada a 4 mujeres entre 61 y 68 años de edad, cuyos temas se basaron en el bienestar subjetivo, la salud, la participación social y la participación ocupacional en el tiempo libre, elementos relevantes en el envejecimiento positivo. Mediante el análisis de datos con la Teoría Fundamentada, se estudió el significado construido sobre el envejecimiento positivo, concebido por las participantes, como una educación del envejecimiento, con el fin de no depender de otros. También se visualizó el envejecimiento como sinónimo de vejez, que se vivencia como una etapa de soledad, en la que si bien, la calidad de vida no se ve afectada por éste, hay factores que influyen en ella, afectando sus capacidades motoras, por ello desarrollan principalmente ocupaciones de tiempo libre pasivas y deseos de efectuar actividades recreativas.
Population aging has generated an increase of older adults, where a large percentage of these are women. Chronic diseases have also increased, being more common in this population joint pathologies, which impact on functionality and quality of life, contributing to the belief in myths about old age. However, this vision would change, due to the creation of a governmental policy proposal such as Positive Aging, aimed to the inclusion of the elderly in society, improving their quality of life and positioning them as a subject of rights.Occupational therapy promotes occupational participation of older adults with joint diseases, so that they have a positive aging. Though, in order to achieve that purpose, it is necessary to analyze the built meaning that these people have about this concept. For this, a semi-structured interview was conducted to women between 61 and 68 years of age, whose themes were based on subjective well-being, health, social participation and occupational participation in free time, relevant elements in positive aging.Through the analysis of data with the Grounded Theory, the meaning built on positive aging, conceived by the participants, was studied. It was conceived as an education of aging, so as not to depend on others. Aging was also seen as synonymous with old age, which is experienced as a stage of loneliness, in which, although quality of life is not affected by it, there are factors that influence it, affecting their motor skills. For this reason they mainly develop passive leisure activities, with the desire to carry out recreational activities.