A fluorescence-activatable reporter of flavivirus NS2B–NS3 protease activity enables live imaging of infection in single cells and viral plaques

The genus Flavivirus in the family Flaviviridae comprises many medically important viruses, such as dengue virus (DENV), Zika virus (ZIKV), and yellow fever virus. The quest for thera- peutic targets to combat flavivirus infections requires a better understanding of the kinetics of virus–host interactions during infections with native viral strains. However, this is precluded by limitations of current cell-based systems for monitoring flavivi- rus infection in living cells. In the present study, we report the construction of fluorescence-activatable sensors to detect the activities of flavivirus NS2B–NS3 serine proteases in living cells. The system consists of GFP-based reporters that become fluo- rescent upon cleavage by recombinant DENV-2/ZIKV proteases in vitro. A version of this sensor containing the flavivirus inter- nal NS3 cleavage site linker reported the highest fluorescence activation in stably transduced mammalian cells upon DENV-2/ ZIKV infection. Moreover, the onset of fluorescence correlated with viral protease activity. A far-red version of this flavivirus sensor had the best signal-to-noise ratio in a fluorescent Dulbec- co’s plaque assay, leading to the construction of a multireporter platform combining the flavivirus sensor with reporter dyes for detection of chromatin condensation and cell death, enabling studies of viral plaque formation with single-cell resolution. Finally, the application of this platform enabled the study of cell-population kinetics of infection and cell death by DENV-2, ZIKV, and yellow fever virus. We anticipate that future studies of viral infection kinetics with this reporter system will enable basic investigations of virus–host interactions and facilitate future applications in antiviral drug research to manage flavivi- rus infections.International Centre for Genetic Engineering and Biotechnology Grant CRP/CRI18-02.UCR::Vicerrectoría de Investigación::Unidades de Investigación::Ciencias de la Salud::Centro de Investigación en Enfermedades Tropicales (CIET)UCR::Vicerrectoría de Docencia::Salud::Facultad de Microbiologí

Propiedades psicométricas del Cuestionario de Aceptación y Acción-II en adultos ecuatorianos

Objetivo. Analizar las propiedades psicométricas del AAQ-II en población adulta ecuatoriana con tomas presenciales, aumentando la fiabilidad de la muestra. Método. En este estudio se evaluaron las propiedades psicométricas de la Acceptance and Action Questionnaire-II (AAQ-II) en población ecuatoriana. Participaron 450 personas entre 18 y 53 años, de las cuales un 46.7% fueron mujeres y un 53.3% hombres. Resultados. En el análisis factorial confirmatorio se encontró una estructura de tres dimensiones: evitación de recuerdos, evitación emocional y falta de autodominio. Un análisis de invarianza reveló que la escala es invariante entre hombres y mujeres. Se encontró que la escala presenta buena consistencia interna (α = .90). Se discute la validez de la AAQ-II como una herramienta para evaluar la evitación experiencial en población adulta ecuatoriana

A Hartree-Bose mean-field approximation for IBM-3

A Hartree-Bose mean-field approximation for the IBM-3 is presented. A Hartree-Bose transformation from the spherical to the deformed bosons with charge-dependent parameters is proposed which allows bosonic pair correlations and includes higher angular momentum bosons. The formalism contains previously proposed IBM-2 and IBM-3 intrinsic states as particular limits.Dirección General de Investigación Científica y Técnica PB 95/0123, PB95-0533European Commission CI1*-CT94-007

Clinical insights into dermatofibrosarcoma protuberans

Dermatofibrosarcoma protuberans (DFSP) is a rare, locally aggressive cutaneous soft tissue sarcoma, it is the second most common skin sarcoma after Kaposi's sarcoma. The cause of DFSP remains unknown. The case of a 54-year-old female patient with a diagnosis of Dermatofibrosarcoma protuberans is presented, displaying a typical clinical presentation. It is characterized by an initial lesion in the form of a reddish spot on the anterior region of the chest, which showed slow growth until the development of a multinodular and irregular lesion with multiple recurrences. The lesion is resected, confined to the superficial layers of the skin, with 3 cm margins, confirming the histopathological diagnosis of dermatofibrosarcoma protuberans with clear margins. DFSP is an uncommon cutaneous sarcoma that is typically low- to intermediate-grade, and while it has a limited likelihood of metastasis, it exhibits a notable tendency for local recurrence. The risk of recurrence is closely linked to the extent of surgical resection

Hartree-Bose mean-field approximation for the interacting boson model (IBM-3)

A Hartree-Bose mean-field approximation for the IBM-3 is presented. A Hartree-Bose transformation from spherical to deformed bosons with charge-dependent parameters is proposed which allows bosonic pair correlations and includes higher angular momentum bosons. The formalism contains previously proposed IBM-2 and IBM-3 intrinsic states as particular limits.DGICYT PB95/0123 PB95–0533Comisión Europea CI1*-CT94-007

The Proteasomal Deubiquitinating Enzyme PSMD14 Regulates Macroautophagy by Controlling Golgi-to-ER Retrograde Transport

Ubiquitination regulates several biological processes, however the role of specific members of the ubiquitinome on intracellular membrane trafficking is not yet fully understood. Here, we search for ubiquitin-related genes implicated in protein membrane trafficking performing a High-Content siRNA Screening including 1187 genes of the human “ubiquitinome” using amyloid precursor protein (APP) as a reporter. We identified the deubiquitinating enzyme PSMD14, a subunit of the 19S regulatory particle of the proteasome, specific for K63-Ub chains in cells, as a novel regulator of Golgi-to-endoplasmic reticulum (ER) retrograde transport. Silencing or pharmacological inhibition of PSMD14 with Capzimin (CZM) caused a robust increase in APP levels at the Golgi apparatus and the swelling of this organelle. We showed that this phenotype is the result of rapid inhibition of Golgi-to-ER retrograde transport, a pathway implicated in the early steps of the autophagosomal formation. Indeed, we observed that inhibition of PSMD14 with CZM acts as a potent blocker of macroautophagy by a mechanism related to the retention of Atg9A and Rab1A at the Golgi apparatus. As pharmacological inhibition of the proteolytic core of the 20S proteasome did not recapitulate these effects, we concluded that PSMD14, and the K63-Ub chains, act as a crucial regulatory factor for macroautophagy by controlling Golgi-to-ER retrograde transport

Avanzando hacia un curriculum por competencias profesionales en Ingeniería Agronómica

Un rasgo esencial de las competencias es la relación entre práctica-teoría, en la cual la práctica delimita la teoría acorde a condiciones y demandas de situaciones concretas en un contexto dado. En las Ciencias Agrarias hay consenso que el sustento de la formación profesional está en la interdisciplinariedad, por tanto se plantea la necesidad de abordar el estudio de los agrosistemas desde equipos multidisciplinarios. El objetivo de este trabajo es evaluar la experiencia “Proyecto cursos correlativos por crédito12: Integración de conocimientos a partir de la articulación teoría-práctica desde la Física al manejo conservacionista del cultivo” – Edición 2010 y 2011–. En la misma participaron alumnos que habían regularizado la asignatura Mecanización Agrícola de la carrera Ingeniería Agronómica y docentes de las cátedras de Física, Topografía, Edafología, Mecanización Agrícola, Manejo de Suelo y Riego y Cultivos Industriales; desarrollándose actividades de integración entre las áreas curriculares de la carrera. Los resultados fueron analizados desde la percepción del alumno mediante encuestas y desde los docentes mediante el análisis de las mismas, la observación participante, registros y la reflexión grupal; de lo cual surge que esta metodología didáctica innovadora, permite avanzar hacia una formación en competencias profesionales.Eje temático 1: Problemáticas y alternativas de mejora de la enseñanza e - Los enfoques y modalidades de integración curricularFacultad de Ciencias Agrarias y Forestale