A two-step lyssavirus real-time polymerase chain reaction using degenerate primers with superior sensitivity to the fluorescent antigen test

A generic two-step lyssavirus real-time reverse transcriptase polymerase chain reaction (qRT-PCR), based on a nested PCR strategy, was validated for the detection of different lyssavirus species. Primers with 17 to 30% of degenerate bases were used in both consecutive steps. The assay could accurately detect RABV, LBV, MOKV, DUVV, EBLV-1, EBLV-2, and ABLV. In silico sequence alignment showed a functional match with the remaining lyssavirus species. The diagnostic specificity was 100% and the sensitivity proved to be superior to that of the fluorescent antigen test. The limit of detection was <= 1 50% tissue culture infectious dose. The related vesicular stomatitis virus was not recognized, confirming the selectivity for lyssaviruses. The assay was applied to follow the evolution of rabies virus infection in the brain of mice from 0 to 10 days after intranasal inoculation. The obtained RNA curve corresponded well with the curves obtained by a one-step monospecific RABV-qRT-PCR, the fluorescent antigen test, and virus titration. Despite the presence of degenerate bases, the assay proved to be highly sensitive, specific, and reproducible

Branding corporativo para una nueva línea de ropa semi formal en la ciudad de Cuenca para mujeres de 25 a 30 años de nivel socioeconómico medio-alto

Vivimos en un mundo con una enorme oferta comercial y no es raro escuchar que algunas marcas desaparecen del mercado. Este estudio de investigación argumenta la importancia del branding corporativo al momento de crear una marca y su influencia en el comportamiento de compra basado en los vínculos emocionales. Tiene por objeto realizar una propuesta de branding que sirva como guía para nuevas tiendas de ropa, teniendo como resultado la creación y posicionamiento de marca. La metodología utilizada es cualitativa y cuantitativa; se realizó encuestas a mujeres de 25 a 30 de nivel socioeconómico medio-alto en la ciudad de Cuenca y entrevistas a empresarias locales propietarias de las marcas Clodette y El- La Clothing. En base a los resultados, se propone la creación de la marca Praga bajo evidencias factibles para el desarrollo de marca, tomando en cuenta las necesidades de las clientas potenciales de clase media de la ciudad de Cuenca. Además, el presente trabajo contribuye con criterios sobre branding adaptados al mercado de mujeres cuencanas, por lo cual este trabajo contribuye con aportes fundamentales para la gestión de marcas de ropa que exploran las estrategias de branding de moda y el manejo estratégico de las redes sociales como herramienta de promoción.We live in a world with huge commercial offers, and it is not uncommon to hear that some brands disappear from the market. This research study argues the importance of corporate branding when creating a brand and its influence on purchasing behavior based on emotional ties. This project aimed to develop a branding proposal that serves as a guide for new clothing stores for its brand creation and positioning. The methodologies used are qualitative and quantitative. Surveys were conducted with women from 25 to 30 of medium-high socioeconomic strata in the city of Cuenca and interviews with local businesswomen owners of the Clodette and El-La Clothing brands. Based on the results, the creation of the brand Praga is proposed under the study ́s results with demonstrate feasible evidence for brand development, taking into account the needs of potential middle-class female customers in the city of Cuenca. In addition, this work contributes with criteria on branding adapted to the market of women from Cuenca, so this work presents fundamental contributions for the management of clothing brands that explore fashion branding strategies and the strategic management of social networks as a promotional tool.Licenciado en Ciencias de la Comunicación Social en Comunicación Organizacional y Relaciones PúblicasCuenc

Prevalence of prematurity and risk factors in newborns admitted to the neonatology area, Hospital Vicente Corral Moscoso, 2011-2015

La prematuridad, a nivel mundial constituye la primera causa de mortalidad y morbilidad neonatal, siendo por ello un verdadero reto para el sistema de salud Ecuatoriano, el objetivo principal de la investigación es el establecimiento de la prevalencia de prematuridad y sus factores asociados, en el área de Neonatología del Hospital Vicente Corral Moscoso, entre los años 2011-2015, aplicando una metodología analística, transversal observacional, revisando un número de 447 historias clínicas de neonatos, los cuales cumplieron los criterios de inclusión, analizando estadísticamente, reportándose Odds ratio, con un intervalo de confianza del (95%) y aceptando el pi value, se aplicaron los fundamentos bioéticos pertinentes, dentro de los resultados se da que la prevalencia de prematuridad fue del (29,4%), el porcentaje de madres adolescentes fue de (30,3%), el (56,1%) posee instrucción secundaria, (36,4%) son casadas y un (72,7%) pernoctan en zonas urbanas. El sexo predominante de los prematuros fue masculino con (52,3%), el (70,5%) fueron prematuros tardíos y el 79,5% pequeños para la edad gestacional.&nbsp; Los factores de riesgo que muestran relación variación estadística fueron: edad menor a 19 años y mayor 35 años ( OR:1.71 ; IC: 1.12-2.59 p: 0.01), trastornos hipertensivos (OR:1.81; IC: 1.08-3.03 p: 0.02), restricción de crecimiento intrauterino (OR:4.89 IC:3.17-7.55 p: 0.000), y embarazo múltiple (OR: 2.79; IC: 1.45-5.34 p: 0.001), concluyendo que la prevalencia de prematuridad fue elevada, se encontró relación estadísticamente significativa con: edad, trastornos hipertensivos, retardo del crecimiento intrauterino y embarazo múltiplePrematurity, worldwide, is the leading cause of neonatal mortality and morbidity, making it a real challenge for the Ecuadorian health system. The main objective of the research is to establish the prevalence of prematurity and its associated factors, in the Neonatology area of ​​the Vicente Corral Moscoso Hospital, between 2011-2015, applying an analytical, cross-sectional observational methodology, reviewing a number of 447 clinical records of newborns, who met the inclusion criteria, statistically analyzing, reporting Odds ratio, with a confidence interval of (95%) and accepting the pi value, the relevant bioethical foundations were applied, within the results it is given that the prevalence of prematurity was (29.4%), the percentage of adolescent mothers was ( 30.3%), (56.1%) have secondary education, (36.4%) are married and (72.7%) spend the night in urban areas. The predominant sex of premature infants was male with (52.3%), (70.5%) were late preterm infants and 79.5% were small for gestational age. The risk factors that show a statistical variation relationship were: age younger than 19 years old and older than 35 years old (OR: 1.71; CI: 1.12-2.59 p: 0.01), hypertensive disorders (OR: 1.81; CI: 1.08-3.03 p: 0.02 ), intrauterine growth restriction (OR: 4.89 CI: 3.17-7.55 p: 0.000), and multiple pregnancy (OR: 2.79; CI: 1.45-5.34 p: 0.001), concluding that the prevalence of prematurity was high, a relationship was found statistically significant with: age, hypertensive disorders, intrauterine growth retardation and multiple pregnanc

Clinical burden of hepatitis E virus infection in a tertiary care center in Flanders, Belgium

Background: Hepatitis E virus (HEV) infection is increasingly recognized as a cause of hepatitis in developed countries. A high HEV IgG seroprevalence in humans and pigs is reported as well as sporadic clinical cases of autochtonous HEV but there are currently no data available on the clinical burden of HEV in Belgium. Objectives: The objective of the current study was to evaluate the actual clinical burden of HEV infections in our tertiary care center in Flanders, Belgium. Study design: In the setting of Ghent University Hospital, patients were assessed for the presence of HEV IgG and IgM as well as HEV RNA if no other cause was found for one of the following clinical presentations: a) elevation of liver enzymes in post-liver transplant; b) suspicion of acute or toxic hepatitis; c) unexplainable elevation of liver enzymes; d) cirrhosis with acute-on-chronic exacerbation. Results: In a period of 39 months (January 2011-April 2014) 71 patients were enrolled. HEV IgG was found positive in 13 (18,3%) patients; HEV IgM in 6 patients (8,5%) and HEV RNA in 4 (5,6%) patients. All HEV IgM/ RNA positive patients were male, aged 41-63, and classified in the clinical groups a), b) or d). HEV IgG seroprevalence was slightly higher but not significantly different from the seroprevalence in the general population in this region in Belgium previously reported to be 14% (p-value 0.41) by our group. Conclusions: HEV should be considered as a cause of liver pathology especially in middle-aged men with elevation of liver enzymes

Etude de la mort cellulaire induite par une réponse antivirale ou un stress du réticulum endoplasmique dans les lignées lymphoïdes

Cancer is caused by the uncontrolled proliferation of aberrant cells that harbour multiple oncogenic mutations in signaling pathways implicated in growth and cell death. Most of today’s anticancer therapies lead to destruction of tumour cells by apoptosis. However, mutations leading to the development of cancer not only promote tumour progression, but often allow cells to avoid apoptotic death and thus result in resistance to treatment. Fortunately, many pathways lead to apoptosis and alternative cell death programmes do exist. Advances in the knowledge about how apoptosis and alternative cell death are activated are important and could pave the way for the development of alternative therapies. Using lymphoid cell lines, in particular Jurkat T cells, we have studied cell death pathways induced by an antiviral response or a prolonged endoplasmic reticulum (ER) stress. First, we have studied the involvement of RIP1 and the activation of dsRNA-activated protein kinase (PKR) in external dsRNA-induced apoptotic and necrotic cell death in Jurkat T cell lymphoma. Our findings suggest that RIP1 is implicated in apoptosis and necrosis induced by dsRNA and a caspase-dependent mode of activation of PKR in apoptosis. PKR is cleaved in two fragments, PKR-ND containing the N-terminal domain of PKR and PKR-KD containing the kinase domain. The PKR-KD fragment interacts with and activates intact PKR. PKR-ND facilitates the interaction of PKR-KD with full-length PKR and thus the activation of the kinase and amplifies the translation inhibitory signal. Secondly, we explored the pathways leading to caspase activation and the involvement of Bcl2 pro-apoptotic members and RIP1 in cell death induced by ER stress in Jurkat T cells. Our results show that caspases-3, -8 and -9 are implicated in apoptosis induced by brefeldin-A (BFA). Using caspase-8 and FADD deficient Jurkat T cells, we demonstrate that these proteins are redundant for BFA induced apoptosis. In Bcl2 overexpressing Jurkat cells exposed to BFA, survival was tremendously improved, although a fraction of the cells died by necrosis. Interestingly, caspase-8 deficient cells were more resistant to thapsigargin and Bcl2 overexpression had no effect on the cytotoxicity of this inhibitor, suggesting that BFA and thapsigargin induce distinct cell death signaling pathways. Comparing Bcl2-like proteins expression in BFA induced apoptotic with necrotic cells; we demonstrate significant 9 differences suggesting that different Bcl2 like proteins may play a role in distinct ER stress induced cell death pathways. To conclude these results bring new data about apoptotic and necrotic pathways induced by an antiviral response and ER stress in lymphoid cell lines. Alternative cell death pathways induced by dsRNA or ER stress could be interesting pathways in developing new cancer therapies.Le cancer est causé par la prolifération incontrôlée de cellules suite à de nombreuses altérations génétiques affectant notamment les voies de signalisation impliquées dans la croissance et la mort cellulaires. La majorité des thérapies anticancéreuses visent à induire l’apoptose des cellules cancéreuses, cependant, les cellules y deviennent de plus en plus résistantes. Heureusement, le concept d’apoptose ne représente pas le seul mécanisme pouvant mener à la mort cellulaire et d’autres types de mort émergent et doivent être pris en considération. Une meilleure compréhension des mécanismes d’apoptose et des modes alternatifs de mort cellulaire permettrait de déterminer de nouvelles cibles dans le but d’améliorer certaines thérapies actuelles. Nos recherches ont porté sur l’étude de la mort cellulaire induite lors d’une réponse antivirale ou lors d’un stress du réticulum endoplasmique (RE) dans des lignées lymphoïdes et plus particulièrement, les cellules T Jurkat. D’une part, nous nous sommes intéressés à décortiquer les mécanismes de l’activation de la kinase PKR (double stranded RNA-dependent kinase) par les caspases et à l’implication de RIP1 au cours de l’apoptose et de la nécrose induites par l’ARNdb (ARN double brin) dans les cellules T Jurkat. Nos résultats suggèrent que RIP1 est impliqué dans l’apoptose et la nécrose induite par l’ARNdb. Nous mettons aussi en évidence un nouveau mode d’activation de PKR dépendant des caspases au cours de l’apoptose. La PKR est clivée par les caspases en fragments N-terminal (PKR-ND) et C-terminal porteur du domaine kinase (PKR-KD). Ces deux fragments montrent des fonctions différentes. PKR-KD est capable de se lier et d’activer la PKR tandis que PKR-ND amplifie ce processus en facilitant l’interaction entre PKR-KD et la kinase PKR entière. D’autre part, nous avons étudié l’implication des caspases, des membres de la famille Bcl-2 et de RIP1 (receptor-interacting protein-1) dans l’apoptose et la nécrose induites par un stress du RE. Nous avons montré que les caspases-3, -8 et -9 jouent un rôle dans l’apoptose induite par la bréfeldine-A (BFA). Par l’utilisation de cellules Jurkat déficientes en caspase-8 et en FADD, nous avons montré que ces protéines sont redondantes dans ce processus apoptotique. Par contre, une surexpression de Bcl-2 augmente clairement la survie cellulaire et les cellules mourantes meurent par nécrose. De plus, les cellules Jurkat déficientes en caspase-8 semblent plus résistantes à la thapsigargine tandis que la surexpression de Bcl-2 n’a pas d’effet sur la cytotoxicité liée à cet inhibiteur, ce qui suggère que la BFA et la thaspigargine induisent des voies de signalisation de mort cellulaire différentes. En comparant l’abondance des protéines pro-apoptotiques de la famille Bcl-2 dans l’apoptose et la nécrose induite par la BFA, nous avons mis en évidence des différences de profil d’expression suggérant que ces protéines jouent un rôle dans la mort cellulaire induite par un stress du RE. En conclusion, les résultats obtenus apportent de nouvelles données dans la compréhension des mécanismes apoptotiques et nécrotiques induits par une réponse antivirale ou de stress du RE dans les lignées lymphoïdes. Les voies alternatives de mort cellulaire induite par l’ARNdb, plus particulièrement la PKR, et les voies induites par un stress du RE semblent des pistes intéressantes afin d’améliorer certains traitements contre le cancer(DOCSC03) -- FUNDP, 201

Subtype-specific differences in the risk of hospitalisation among patients infected with hepatitis E virus genotype 3 in Belgium, 2010-2018

Some European countries recently reported an increase in hepatitis E virus genotype 3 (HEV-3) of the subtype 3c. No link between HEV-3 subtypes and severity is established to date. Here, we report that patients infected with HEV-3c were at lower risk of hospitalisation, compared to those infected with HEV-3f, the other main subtype circulating in Belgium

Prevalencia de prematuridad y factores de riesgo asociados en los recién nacidos, ingresados en el área de neonatología del Hospital Vicente Corral Moscoso entre el periodo 2011 al 2015

Antecedentes: la prematuridad a nivel mundial es la primera causa de mortalidad y morbilidad neonatal siendo por ello un verdadero reto para los sistemas de salud. Objetivo general: establecer la prevalencia de prematuridad y sus factores asociados, en el Área de Neonatología del Hospital Vicente Corral Moscoso (HVCM) entre el periodo 2011 al 2015. Metodología: estudio observacional, analítico, transversal. Se revisó 447 historias clínicas de neonatos del HVCM que cumplieron los criterios de inclusión. Los datos fueron analizados mediante programa SPSS 15, y presentados en tablas personalizadas. Para establecer la relación con factores de riesgo se aplicó la prueba del chi cuadrado, reportándose odds ratio, intervalo de confianza al 95% y valor de p. Se aplicaron los fundamentos bioéticos pertinentes. Resultados: la prevalencia de prematuridad fue de 29.4%, 30.3% de las madres fueron adolescentes, el 56.1%, con instrucción secundaria, 36.4%, casadas, y un 72.7%, de zonas urbanas. El sexo predominante de los prematuros fue hombre, con 52.3%, el 70.5% fueron prematuros tardíos y 79.5% pequeños para la edad gestacional. Los factores de riesgo que mostraron relación estadísticamente significativa fueron: edad menor a 19 y mayor 35 años ( OR:1.71 ; IC: 1.12-2.59 p: 0.01), trastornos hipertensivos (OR:1.81; IC: 1.08-3.03 p: 0.02), restricción de crecimiento intrauterino (OR:4.89 IC:3.17-7.55 p: 0.000), y embarazo múltiple (OR: 2.79; IC: 1.45-5.34 p: 0.001). Conclusiones: la prevalencia de prematuridad fue elevada, se encontró relación estadísticamente significativa con: edad, trastornos hipertensivos, retardo del crecimiento intrauterino y embarazo múltipleBackground: Prematurity worldwide is the leading cause of neonatal mortality and morbidity, which is why it is a real challenge for health systems. General Purpose: Establish the prevalence of prematurity and factors associated with prematurity in the Area of Neonatology of the Hospital Vicente Corral Moscoso (HVCM) between the period 2011 to 2015. Methodology: Observational, analytical, cross-sectional study. We reviewed 447 clinical histories of HVCM neonates that met the inclusion criteria. The data was analyzed through the SPSS 15 program and presented in personalized tables. To establish the relationship with risk factors, the chi square test was applied, reporting odds ratio, 95% confidence interval and p value. The pertinent bioethics foundations are applied. Results: The prevalence of prematurity was 29.4%, 30.3% of mothers of premature babies were teenagers, 56.1% of high school, 36.4% married and 72.7% of urban areas. The predominant sex of preterm infants was males with 52.3%, 70.5% late preterm and 79.5% small for gestational age. The risk factors that showed a statistically significant relationship were: age less than 19 and greater than 35 years (OR: 1.71, CI: 1.12-2.59 p: 0.01), hypertensive disorders (OR: 1.81, CI: 1.08-3.03 p: 0.02), intrauterine growth restriction (OR: 4.89 CI: 3.17-7.55 p: 0.000), and multiple pregnancy (OR: 2.79, CI: 1.45-5.34 p: 0.001). Conclusion: The prevalence of prematurity was high, a statistically significant relationship was found with: age, hypertensive disorders, intrauterine growth retardation and multiple pregnancyMédicoCuenc