Escherichia coli as a New Platform for the Fast Production of Vault-like Nanoparticles: An Optimized Protocol.

Vaults are protein nanoparticles that are found in almost all eukaryotic cells but are absent in prokaryotic ones. Due to their properties (nanometric size, biodegradability, biocompatibility, and lack of immunogenicity), vaults show enormous potential as a bio-inspired, self-assembled drug-delivery system (DDS). Vault architecture is directed by self-assembly of the "major vault protein" (MVP), the main component of this nanoparticle. Recombinant expression (in different eukaryotic systems) of the MVP resulted in the formation of nanoparticles that were indistinguishable from native vaults. Nowadays, recombinant vaults for different applications are routinely produced in insect cells and purified by successive ultracentrifugations, which are both tedious and time-consuming strategies. To offer cost-efficient and faster protocols for nanoparticle production, we propose the production of vault-like nanoparticles in Escherichia coli cells, which are still one of the most widely used prokaryotic cell factories for recombinant protein production. The strategy proposed allowed for the spontaneous encapsulation of the engineered cargo protein within the self-assembled vault-like nanoparticles by simply mixing the clarified lysates of the producing cells. Combined with well-established affinity chromatography purification methods, our approach contains faster, cost-efficient procedures for biofabrication in a well-known microbial cell factory and the purification of "ready-to-use" loaded protein nanoparticles, thereby opening the way to faster and easier engineering and production of vault-based DDSs.

Cinco años en el diagnóstico prenatal de cardiopatías congénitas en Las Tunas / Five years of the prenatal diagnosis of congenital heart diseases in Las Tunas

Fundamento: las cardiopatías congénitas constituyen uno de los defectos más frecuentes en el ser humano, su diagnóstico prenatal es posible en más de un 80 por ciento.Objetivo: caracterizar el diagnóstico prenatal de cardiopatías congénitas en Las Tunas, durante los años 2011 a 2015.Métodos: estudio descriptivo y retrospectivo de corte transversal realizado en el departamento de genética provincial, Hospital Pediátrico Provincial Mártires de Las Tunas. El universo fueron las 31630 gestantes seguidas por riesgo genético de cardiopatía congénita y la muestra las 115 con diagnóstico confirmado. Se analizaron: municipio de procedencia, edad materna, cardiopatía según defecto anatómico, decisiones de interrupciones y seguimientos e incidencia en la mortalidad infantil.Resultados: los municipios de Las Tunas y Puerto Padre presentaron mayor número de diagnóstico prenatal de cardiopatías congénitas; el grupo de edad de 20 a 35 años mostró mayor incidencia con 87 casos, para un 75,7 por ciento; las cardiopatías representaron el 22 por ciento entre todos los defectos congénitos; la tetralogía de Fallot (18), síndrome de hipoplasia de corazón izquierdo (16) y la comunicación interventricular (14) fueron las más diagnosticadas. A las cardiopatías congénitas complejas se le realizaron interrupción del embarazo (89), confirmándose por anatomía patológica el diagnóstico en 83 casos, para un 92,3 por ciento.Conclusiones: el diagnóstico prenatal oportuno de las cardiopatías, unido al adecuado asesoramiento genético, ha incidido positivamente en la disminución de la tasa de la mortalidad por estas causas y en la tasa de mortalidad infantil (AU)

Background: congenital heart diseases are one of the most frequent defects in human beings. Its prenatal diagnosis is possible in more than 80 percent.Objective: to characterize the prenatal diagnosis of congenital heart diseases in Las Tunas, from 2011 through 2015.Methods: a descriptive and retrospective cross-sectional study was carried out at the provincial department of genetics of Mártires de Las Tunas Provincial Pediatric Hospital. The universe included the 31630 pregnant women followed-up due to genetic risk for congenital heart disease. The sample was made up of the 115 patients with a confirmed diagnosis. The following indicators were analyzed: municipality of origin, maternal age, cardiopathy according to anatomical defect, termination decisions, as well as follow-up and incidence in infant mortality.Results: the municipalities of Las Tunas and Puerto Padre presented a higher number of prenatal diagnoses of congenital heart diseases. The 20 to 35 age group showed a higher incidence with 87 cases, for 75,7 percent. Cardiopathies represented a 22 percent of all congenital anomalies. Tetralogy of Fallot (18), hypoplastic left heart (16) and interventricular communication (14) were the conditions mostly diagnosed. The patients with complex congenital cardiopathies underwent termination of pregnancy (89). The diagnosis was confirmed by pathological studies in 83 cases, for 92,3 percent.Conclusions: early prenatal diagnosis of heart diseases, together with an adequate genetic counseling, has had a positive effect on the decline in the mortality rate due to these causes and in the infant mortality rate (AU)