Seroprevalence of Zika virus among blood donors before the epidemic in Barranquilla, Colombia, 2015-2016.

The first cases of Zika virus infection in Colombia were reported and confirmed in October 2015. The objective of the study was estimate the seroprevalence of ZIKV infection during the pre-epidemic phase in Barranquilla, Colombia, and demonstrate the presence of virus before the Colombian Ministry of Health confirmed the first case. We conducted a descriptive study of the seroprevalence of Zika virus in 390 samples obtained from a blood bank located in Barranquilla, Colombia - a city endemic for dengue, and with a recent history of a Chikungunya disease epidemic. The serum pools were tested using Euroimmun ZIKV ELISA kit. Seroprevalence of Zika virus IgG were: May 2015: 0%, June and July 2015: 2.62% (95% CI = 0.28-12.13) and August 2015: 5.35% (95% CI = 1.74-16.74). This brings to our attention the need for extending the surveillance period of this virus in order to adequately assess its teratogenic effects.

Evaluation of a loop-mediated isothermal amplification method for rapid detection of human respiratory syncytial virus in children with acute respiratory infection / Evaluación de un método de amplificación isotérmica medida por bucle para la detección rápida del virus sincitial respiratorio en niños con infección respiratoria aguda

Introduction: Human respiratory syncytial virus (hRSV) is the most frequent cause of acute respiratory infection of the lower respiratory tract in children under the age of five. The development of molecular techniques able to identify hRSV is one of the current challenges in the field of clinical research. Objective: To evaluate the ability of an isothermal amplification method to rapidly detect hRSV in children with acute respiratory infection. Materials and methods: We collected 304 nasopharyngeal swab samples from children with symptoms of acute respiratory infection who attended the emergency unit at Hospital de la Universidad del Norte in Barranquilla from April, 2016, to July, 2017. After extracting viral RNA from the samples, we evaluated the ability of the reverse transcriptase-loop-mediated isothermal amplification (RT-LAMP) M assay to rapidly detect hRSVA and hRSVB compared to other molecular techniques: quantitative PCR (qPCR), reverse transcriptase-LAMP L assay, and as a standard, the multiplex nested reverse transcriptase polymerase chain reaction (nested RT-PCR). Results: The RT-LAMP M assay had a sensitivity of 93.59% and a specificity of 92.92%, and a concordance of 0.83 ± 0.036 as compared with the nested RT-PCR test. While the Kappa index of the RT-LAMP M assay was higher than the values for the RT-LAMP L assay and the qPCR, the values of the latter two methods were in agreement (0.75 ± 0.043 and 0.71 ± 0.045, respectively). Conclusion: Due to the shorter running times, lower costs and better performance of the RT-LAMP M assay, it can be considered as a useful clinical tool for the detection of RSVA.

Introducción. El virus sincicial respiratorio humano (hRSV) es la causa más frecuente de infección respiratoria aguda de las vías respiratorias inferiores en niños menores de cinco años. El desarrollo de técnicas moleculares para identificarlo es uno de los retos actuales en el campo de la investigación clínica. Objetivo. Evaluar un método de amplificación isotérmica para la detección rápida del hRSV en niños con infección respiratoria aguda. Materiales y métodos. Se extrajo el ARN viral de 304 muestras de hisopado nasal en niños con síntomas de infección respiratoria aguda atendidos en el servicio de urgencias del Hospital de la Universidad del Norte en Barranquilla entre abril del 2016 y julio del 2017. Se evaluó la prueba de amplificación isotérmica mediada por bucle mediante transcriptasa inversa de la proteína de la matriz (M) (Reverse Transcription Loop-Mediated Isothermal Amplification, RT-LAMP) comparada con técnicas moleculares como la reacción en cadena de la polimerasa mediante transcriptasa inversa múltiple anidada (Reverse Transcription-Polymerase Chain Reaction, RT-PCR), la cual se empleó como la prueba estándar, la PCR en tiempo real (quantitative PCR, qPCR) y la RT-LAMP de la proteína L (L) para la detección rápida del virus sincicial respiratorio (VSR), subtipo A y subtipo B. Resultados. La prueba de RT-LAMP (M) tuvo una sensibilidad de 93,59 %, una especificidad de 92,92 % y una concordancia de 0,83 ± 0,036 comparada con la prueba de RT-PCR anidada. El índice kappa del RT-LAMP (M) fue superior, y los valores del RTLAMP (L) y la qPCR concordaron (0,75 ± 0,043 y 0,71 ± 0,045, respectivamente).

Diarylethenes Display In Vitro Anti-TB Activity and Are Efficient Hits Targeting the Mycobacterium tuberculosis HU Protein.

Tuberculosis continues to be a great source of concern in global health because of the large reservoir of humans infected with the bacilli and the appearance of clinical isolates resistant to a wide array of anti-tuberculosis drugs. New drugs with novel mechanisms of action on new targets are urgently required to reduce global tuberculosis burden. Mycobacterium tuberculosis nucleoid associated protein (NAP) HU has been shown to be druggable and essential for the organism's survival. In this study, four diarylethenes were synthesized using a one-pot decarboxylated Heck-coupling of coumaric acids with iodoanisoles. The prepared compounds 1-4 were tested for their in vitro growth inhibition of M. tuberculosis H37Rv using the spot culture growth inhibition assay, displaying minimum inhibitory concentrations between 9 and 22 µM. Their cytotoxicity against BHK-21 cell line showed half inhibition at concentrations between 98 and 729 µM. The most selective hit (SI = 81), demonstrated inhibition of M. tuberculosis HU protein involved in maintaining bacterial genome architecture.

ERK-1/2 activity is required for efficient RSV infection.

Respiratory syncytial virus (RSV) infection up-regulates the expression of genes encoding proinflammatory mediators in bronchial epithelial cells. However, the specific signaling events immediately following RSV exposure are poorly understood. Herein, we report that RSV attachment to A549 cells activates both ERK-1 and ERK-2 pathways within 5 min. Inhibition of ERK pathways significantly decreases RSV infection of these cells compared to controls. These results demonstrate that the activation of the ERK-1/2 is required in RSV-induced early gene expression.

Mechanism of bronchoprotective effects of a novel natriuretic hormone peptide.

BACKGROUND: The natriuretic hormone peptide (NHP)(99-126), a C-terminal peptide of pro-atrial natriuretic factor (proANF), induces bronchodilatory effects in people with asthma. Recently, another plasmid-encoded C-terminal peptide, pNHP(73-102), was shown to induce a long-lasting bronchoprotective effect in a mouse model of allergic asthma. OBJECTIVE: This study was carried out to determine the role of lung epithelial cells in the bronchoprotective and anti-inflammatory activity of these peptides. METHODS: Human type II alveolar epithelial cells (A549) and normal human bronchial epithelial (NHBE) cells were transfected with pNHP(73-102) to test the effect of this peptide on activation of these cells. After transfection, cells were analyzed for changes in Ca(++) and nitric oxide (NO) levels. Also, activation of NFkappaB and the extracellularly regulated kinase (ERK) 1, 2 signaling pathway was examined by luciferase reporter assay and phosphorylation studies respectively. RESULTS: Analysis of intracellular Ca(++) levels in pNHP(73-102) -transfected A549 or NHBE showed that the peptide increases release. This Ca(++) release was accompanied by an increase in the production of NO. Also, overexpression of pNHP(73-102), but not pVAX control, in phorbol myristate acetate-activated A549 cells resulted in a significant decrease in expression of a cotransfected nuclear factorkappaB (NFkappaB)-luciferase reporter. Similarly, pNHP(73-102) decreased TNF-alpha-induced NFkappaB activation in NHBE cells. Furthermore, NHP(73-102) but not atrial natriuretic peptide decreased phosphorylation of Erk-1, 2 in A549 cells. CONCLUSIONS: Overexpression of pNHP(73-102) in epithelial cells causes increased production of intracellular Ca(++) and NO, with a concomitant decrease in activation of NFkappaB and ERK1, 2. These results suggest a bronchodilatory and anti-inflammatory activity of this peptide.

Respiratory syncytial virus infection activates STAT signaling in human epithelial cells.

Acute respiratory syncytial virus (RSV) infection causes airway inflammation and exacerbates asthma, but the mechanism of inflammation is poorly understood. The role of the STAT-signaling pathway in RSV infection in epithelial cells was examined in this study. DNA microarray analyses of RSV-infected human alveolar type II (A549) epithelial cells identified several genes whose expression was altered from -5.5 to +56.4-fold. Four of the highly expressed genes contained STAT-binding elements. In A549 and normal human bronchial epithelial cells (NHBE), RSV induced phosphorylation and nuclear translocation of STAT-1alpha that was abrogated when RSV attachment was blocked. Treatment with a JAK-2 inhibitor or transfection with dominant-negative STAT-1alpha blocked STAT-1alpha activation and RSV infection. RSV also activated STAT-3 and IL-6 specific antibodies blocked this activation. Thus, activation of the STAT-1alpha and STAT-3 pathways play a role in RSV infection.

Vaccine development against HIV-1: current perspectives and future directions.

The development of an efficacious vaccine against the human immunodeficiency virus (HIV) is of great urgency, because it is accepted that vaccination is the only means capable of controlling the AIDS pandemic. The foundation of HIV vaccine development is the analysis of immune responses during natural infection and the utilization of this knowledge for the development of protective immunization strategies. Initial vaccine development and experimentation are usually in animal models, including murine, feline, and nonhuman primates. Experimental vaccine candidates are closely studied for both efficacy and safety before proceeding to human clinical trials. There are a number of different therapeutic and prophylactic vaccine strategies currently being studied in human clinical trials. Vaccine strategies that are being tested, or have previously been tested, in humans include subunit, DNA plasmid, and viral vector, and combinations of these various strategies. Some of the results of these trials are promising, and additional research has focused on the development of appropriate chemical and genetic adjuvants as well as methods of vaccine delivery to improve the host immune response. This review summarizes the vaccine strategies that have been tested in both animal models and human clinical trials.

Hipersensibilidad a los alergenos de la cucaracha en una muestra de pacientes asmaticos de la ciudad de Barranquilla / Hypersensibily to allergens of roach in a sample of asthmatic patients of Barranquilla

Los alergenos provenientes de la Cucaracha (CR), actualmente son considerados como factores importantes en el desencadenamiento y en el incremento de la morbilidad en el Asma Bronquial alérgica. Después de la sensibilización a las fracciones alergénicas de los ácaros del polvo, los detritos de CR son los mayores alergenos involucrados en la inmunopatogenia del asma, en especial aquellos pacientes residentes en áreas urbanas. Los detritos provenientes de diferentes constituyentes de CR, representan un componente importante del polvo de casa, particularmente en los estratos económicos deprimidos. Este estudio se llevó a cabo para establecer la prevalencia y evaluar el significado clínico de la hipersensibilidad a los alergenos de CR, en pacientes asmáticos residentes en la ciudad de Barranquilla. Se estudiaron 218 pacientes provenientes de las diferentes clínicas del Instituto de los Seguros Sociales y de la consulta privada. El 50.4 por ciento del total de pacientes mostraron hipersensibilidad evaluada por la Prueba Epicutánea (Prick Test). Sólo el 14 por ciento de 150 sujetos controles, no atópicos, clínicamente normales, tuvieron prueba cutánea positiva (p<0.05). La mayor prevalencia de hipersensibilidad a CR estuvo en el grupo etáreo de 7 - 12 años (27.3 por ciento) y fue más frecuente en pacientes con asma moderada persistente y asma severa: no se observó asociación estadísticamente significativa entre la sensibilización a CR y los diferentes estratos sociales

Alergenos: relación entre función biológica y alergenicidad / Allergens: the relationship between biological function and allergenicity

Los alergenos se encuentran en muy diversas fuentes, pero, tienen la particularidad de inducir la producción de inmunoglobulina E (IgE) y provocar alergia. No se conoce una característica molecular biológica común entre los alergenos que explique su capacidad alergénica. En los últimos años, se han demostrado o inferido diferentes funciones biológicas en los alergenos, generando nuevas hipótesis sobre el papel de dichas funciones en la actividad alergénica. Con el fin de identificar que funciones biológicas se han demostrado o inferido en los alergenos y analizar su posible influencia en el papel alergénico, se revisaron aproximadamente doscientos alergenos teniendo como base los ya caracterizados por el comité de Nomenclatura de Alergenos de la OMS, como también otros menos caracterizados pero con algunas propiedades fisicoquímicas conocidas. Los alergenos se agruparon de acuerdo con la actividad biológica y se tabularon con su fuente de origen, peso molecular (PM), punto isoeléctrico (pl), frecuencia de reactividad en la población alérgica, potenciales sitios de glicosilación y formación de enlaces disulfuro. Se identificaron 88 alergenos en los que se ha informado alguna actividad biológica. El agrupamiento de estos según la actividad produjo los siguientes resultados: enzimática (47,2 por ciento), inhibición de enzimas (11,3 por ciento), transporte (18,1 por ciento), regulación de la actividad celular (15,9 por ciento) y otras actividades como la de conferir resistencia a enfermedades en plantas y citólisis (7,9 por ciento). En muchos de estos alergenos, la actividad biológica ha sido inferida por su homología estructural con proteínas de función conocida, pero los experimentos que corroboren dicha función no se han realizado. La mayoría de estos alergenos tienen en común PM<60 kDa, pl<7,0 y escasa formación de oligómeros. Nuestro análisis sugiere que la capacidad de inducir alergia no está determinada por ninguna de las funciones biológicas descritas. Aunque la actividad de cisteína-proteasa y serina-proteasa de algunos de los alergenos de los ácaros domésticos han sido señaladas como determinantes en el papel alergénico en dichos alergenos, los datos experimentales no son concluyentes y no permiten atribuir la alergenicidad a la función enzimática