The human adenovirus type 5 E1B 55kDa protein interacts with RNA promoting timely DNA replication and viral late mRNA metabolism.

The E1B 55kDa produced by human adenovirus type 5 is a multifunctional protein that participates in the regulation of several steps during the viral replication cycle. Previous studies suggest this protein plays an important role in postranscriptional regulation of viral and cellular gene expression, as it is required for the selective accumulation of maximal levels of viral late mRNA in the cytoplasm of the infected cell; however the molecular mechanisms that are altered or regulated by this protein have not been elucidated. A ribonucleoprotein motif that could implicate the direct interaction of the protein with RNA was initially predicted and tested in vitro, but the interaction with RNA could not be detected in infected cells, suggesting the interaction may be weak or transient. Here it was determined that the E1B 55kDa interacts with RNA in the context of the viral infection in non-transformed human cells, and its contribution to the adenovirus replication cycle was evaluated. Using recombinant adenoviruses with amino acid substitutions or a deletion in the ribonucleoprotein motif the interaction of E1B 55kDa with RNA was found to correlate with timely and efficient viral DNA replication and viral late mRNA accumulation and splicing.

Plantas con actividad insecticida: una alternativa natural contra mosquitos / Insecticidal activity plants: A natural alternative against mosquitoes. Current status of the topic in the America´s region

Resumen El siguiente trabajo pretende reunir la información existente con respecto la evaluación de la actividad insecticida de plantas contra mosquitos realizados en la región de las Américas en los últimos veinte años. Se utilizó Pubmed Central, SCielo regional y BioOne y se acudió a buscadores como Google y Bing. Como criterio de inclusión se introdujeron las categorías: actividad larvicida, adulticida-repelente, ovicida e inhibidora del desarrollo de aceites esenciales y extractos de plantas en condiciones de laboratorio contra mosquitos en el área de las Américas en un período desde 1995-2015. La especie de mosquito más estudiada ha sido Ae aegypti seguido de Cx. quinquefascitus, Ae. albopictus, Cx. tarsalis, Cx. pipiens, An. albimanus. El país a la vanguardia en estudios sobre esta temática es Brasil seguido de Cuba México, Estados Unidos. Estados Unidos se destaca por la evaluación de productos comerciales registrados a base de plantas con actividad repelente. En 239 plantas se evaluó la actividad larvicida. El 64 % de los artículos revisados estudia este tipo de actividad con aceites esenciales. La repelencia es el acápite más estudiado después de la actividad larvicida, dentro de los que se destacan la evaluación de formulaciones comerciales con principios activos naturales. Con escasa representación, se encuentran los estudios sobre la actividad ovicida e inhibidora del desarrollo. Más de 85 plantas se evaluaron en forma de extractos y el extracto mas evaluado fue el etanólico seguido del metanólico y el acuoso.

Abstract The following paper aims to gather the existing information regarding the evaluation of the insecticidal activity of plants against mosquitoes made in the region of the Americas in the last twenty years. Pubmed Central, SCielo regional and BioOne were used and search engines such as Google and Bing were used. As an inclusion criterion, the categories were introduced: larvicidal, adulticidal-repellent, ovicidal activity and inhibitor of the development of essential oils and extracts of plants under conditions of Laboratory in the area of the Americas in a period from 1995-2015. The most studied mosquito species was Ae aegypti followed by Cx. quinquefascitus, Ae. albopictus and Cx tarsalis, Cx. pipiens, An. albimanus. The country at the forefront in studies on this subject is Brazil followed by Cuba Mexico, United States. The United States stands out by evaluating commercially registered herbal products with repellent activity. The larvicidal activity was evaluated in 239 plants. 64% of the reviewed articles study this type of activity with essential oils. The repellency is the most studied section after the larvicidal activity, among which the evaluation of commercial formulations with natural active principles. With little representation, there are studies on ovicidal and developmental inhibitory activity. More than 85 plants have been evaluated in the form of extracts and the most evaluated extract has been ethanolic followed by methanolic and aqueous.

Caracterización parcial de la actividad de esterasas en una cepa de Aedes aegypti resistente a temefos / Partial characterization of esterase activity in a temephos-resistant Aedes aegypti strain

Introducción: las enzimas esterasas han sido identificadas como mecanismo de resistencia a temefos en Aedes aegypti de Cuba, larvicida más utilizado en el mundo. Objetivo: caracterizar parcialmente la actividad de esterasas en larvas expuestas y no expuestas a dosis subletales de temefos en una cepa de Aedes aegypti resistente a este insecticida. Métodos: se utilizó una cepa de Aedes aegypti de referencia susceptible (Rockefeller) y otra resistente a temefos (SANtemF11). Se expusieron las larvas de la cepa SANtemF11 a la concentración letal 90 (CL90) de temefos (1 ppm), 10 % de larvas sobrevivientes a las 24 h (SANtem [24 h]) se transfirieron a agua limpia y sin exposición a insecticidas por otras 24 h (SANtem [48 h]). Se caracterizó de modo parcial, en estas larvas, la actividad de esterasas a través de ensayos bioquímicos y electroforesis en gel de poliacrilamida. Se estimó por duodecil sulfato de sodio (SDS-PAGE) el peso molecular de la esterasa (Est. A4). Resultados: la actividad de esterasas en la cepa SANtemF11 resultó significativamente mayor que en Rockefeller. Se observó una disminución significativa de la actividad de esterasas en las larvas sobrevivientes (SANtemF11 [24 h]), la cual se recuperó 24 h después sin exposición a temefos. En el zimograma se observó que en 10 % de las larvas sobrevivientes a temefos, solo apareció incrementada la banda de esterasa A4, en comparación con las observadas en SANtemF11. El peso molecular estimado de la esterasa A4 fue de 58 kDa. Conclusiones: la presencia de una banda específica de esterasa (58 kDa), en las larvas sobrevivientes a la selección con temefos, confirma su papel en la resistencia a este insecticida. Diagnosticar la función de las esterasas en la resistencia a temefos, a través de ensayos bioquímicos, no debe realizarse en larvas expuestas a dosis subletales de este insecticida, para evitar falsos negativos.

Introduction: the esterase enzymes have been defined as the mechanism of resistance to temephos in Aeges aegypti in Cuba, which is the most used larvacide worldwide. Objective: to partially characterize the activity of esterases in exposed and non-exposed larvae at sublethal doses of temephos in an Aedes aegypti strain that is resistant to this product. Methods: a susceptible reference Aedes aegypti strain (Rockefeller) and another temephos-resistant strain (SANtemFII) were used. The larvae from SANtemF11 strain were exposed to lethal concentration 90 (LC90) of temephos (1 ppm); 10 % of the surviving larvae after 24 hours (SANtem[24 h] was moved to clean water, with no exposure to insecticide for 24 hours (SANtem [48 h]). The activity of esterases was partially characterized in these larvae through biochemical assays and gel-polyacrylamide electrophoresis. The molecular weight of esterase A 4 (ESt. A4) was estimated with the support of sodium duodecyl sulophate (SDS-PAGE). Results: the activity of esterases in SANtemF11 was significantly higher than in Rockefeller strain. Significant reduction of the activity of esterases in surviving larvae was observed (SANtemF11 [24 h], but it increased 24 h later without exposure to temephos. The zymogram showed that 10% of larvae that survived from temephos action, just the esterase A4 band increased if compared with those of SAntemF11. The estimated molecular weight of esterase A4 was 58 kDa. Conclusions: the presence of a specific band of esterase (58 kDa) in surviving larvae confirmed the role of these enzymes in insecticidal resistance. The diagnosis of the function of the esterases in resistance to temephos through biochemical tests should not be made in larvae exposed to sublethal doses of this insecticide, in order to avoid false negatives.

[Partial characterization of esterase activity in a temephos-resistant Aedes aegypti strain]. / Caracterización parcial de la actividad de esterasas en una cepa de Aedes aegypti resistente a temefos.

INTRODUCTION: the esterase enzymes have been defined as the mechanism of resistance to temephos in Aeges aegypti in Cuba, which is the most used larvacide worldwide. OBJECTIVE: to partially characterize the activity of esterases in exposed and nonexposed larvae at sublethal doses of temephos in an Aedes aegypti strain that is resistant to this product. METHODS: a susceptible reference Aedes aegypti strain (Rockefeller) and another temephos-resistant strain (SANtemFII) were used. The larvae from SANtemF11 strain were exposed to lethal concentration 90 (LC90) of temephos (1 ppm); 10 % of the surviving larvae after 24 hours (SANtem[24 h] was moved to clean water, with no exposure to insecticide for 24 hours (SANtem [48 h]). The activity of esterases was partially characterized in these larvae through biochemical assays and gel-polyacrylamide electrophoresis. The molecular weight of esterase A 4 (ESt. A4) was estimated with the support of sodium duodecyl sulophate (SDS-PAGE). RESULTS: the activity of esterases in SANtemF11 was significantly higher than in Rockefeller strain. Significant reduction of the activity of esterases in surviving larvae was observed (SANtemF11 [24 h], but it increased 24 h later without exposure to temephos. The zymogram showed that 10% of larvae that survived from temephos action, just the esterase A4 band increased if compared with those of SAntemF11. The estimated molecular weight of esterase A4 was 58 kDa. CONCLUSIONS: the presence of a specific band of esterase (58 kDa) in surviving larvae confirmed the role of these enzymes in insecticidal resistance. The diagnosis of the function of the esterases in resistance to temephos through biochemical tests should not be made in larvae exposed to sublethal doses of this insecticide, in order to avoid false negatives.