Cellular integrin ¿5ß1 and exosomal adam17 mediate the binding and uptake of exosomes produced by colorectal carcinoma cells

Approximately 25% of colorectal cancer (CRC) patients develop peritoneal metastasis, a condition associated with a bleak prognosis. The CRC peritoneal dissemination cascade involves the shedding of cancer cells from the primary tumor, their transport through the peritoneal cavity, their adhesion to the peritoneal mesothelial cells (PMCs) that line all peritoneal organs, and invasion of cancer cells through this mesothelial cell barrier and underlying stroma to establish new metastatic foci. Exosomes produced by cancer cells have been shown to influence many processes related to cancer progression and metastasis. In epithelial ovarian cancer these extracellular vesicles (EVs) have been shown to favor different steps of the peritoneal dissemination cascade by changing the functional phenotype of cancer cells and PMCs. Little is currently known, however, about the roles played by exosomes in the pathogenesis and peritoneal metastasis cascade of CRC and especially about the molecules that mediate their interaction and uptake by target PMCs and tumor cells. We isolated exosomes by sizeexclusion chromatography from CRC cells and performed cell-adhesion assays to immobilized exosomes in the presence of blocking antibodies against surface proteins and measured the uptake of fluorescently-labelled exosomes. We report here that the interaction between integrin 5 1 on CRC cells (and PMCs) and its ligand ADAM17 on exosomes mediated the binding and uptake of CRC-derived exosomes. Furthermore, this process was negatively regulated by the expression of tetraspanin CD9 on exosome

Nuevos aportes sobre alteraciones cognitivas y metabólico-cerebrales por la administración de ketamina en dosis subanestésicas en ratas Rattus norvegicus (estudio preliminar)

La ketamina antagonista del receptor NMDA fue creada en la década del 60 como anestésico disociativo para niños y adultos; rápidamente ocupo un lugar fundamental en medicina veterinaria. Con el tiempo se le atribuyeron otro tipo de utilidades como analgésico, antidepresivo e incluso anticonvulsivante para algún tipo de estado epiléptico refractario. En nuestro laboratorio lo utilizamos como anestésico para la colocación de cánulas intracerebrales en dosis de 70 mg/kg.2 En el caso de su uso como analgésico para dolor de origen central y neuropático la dosis es muy por debajo de la utilizada para llegar al plano quirúrgico. Por lo que es fundamental saber si estas dosis conllevan consecuencias a nivel cognitivo y qué cambios se producen a nivel metabólico en las estructuras cerebrales implicadas en dichos procesos. En trabajos previos demostramos que la ketamina no produce alteración en el patrón de movimientos con dosis inferiores a 5 mg/ kg pero provoca detrimento de la memoria de trabajo asociado a alteraciones metabólicas de la corteza prefrontal e hipocampo. La amígdala es una estructura que forma parte del sistema límbico, está formada por muchos núcleos, se la relaciona con el miedo, la ansiedad y los procesos de memoria.Fil: Guevara, M. A.. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas; ArgentinaFil: Lorenzo, S.. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas; ArgentinaFil: Anselmi, V.. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas; ArgentinaFil: Romanowicz, E.. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas; ArgentinaFil: Cabrera, M.. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas; ArgentinaFil: García Menéndez, Sebastián Marcelo Manuel. Laboratorio de Neurociencias y Psicología Experimental.; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Barrutieta, I.. Universidad del País Vasco; EspañaFil: Baiardi, G.. Universidad Católica de Córdoba; ArgentinaFil: Lafuente, J. V.. Universidad del País Vasco; EspañaFil: Gargiulo, Pascual Angel. Universidad Nacional de Cuyo. Facultad de Ciencias Médicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza; ArgentinaNEUROLATINVET 2019 - VII Congreso Latinoamericano de Neurología Veterinaria, X Encuentro de Neurología Veterinaria del Cono Sur, II Congreso de Neurocirugía VeterinariaMendozaArgentinaAsociación Argentina de Neurología Veterinari

Geographic variation in the sensitivity of recombinant antigen-based rapid tests for chronic trypanosoma cruzi infection

Chagas disease affects 8-11 million people throughout the Americas. Early detection is crucial for timely treatment and to prevent non-vectorial transmission. Recombinant antigen-based rapid tests had high sensitivity and specificity in laboratory evaluations, but no Peruvian specimens were included in previous studies. We evaluated Stat-Pak and Trypanosoma Detect rapid tests in specimens from Bolivia and Peru. Specimens positive by three conventional assays were confirmed positives; specimens negative by two or more assays were confirmed negatives. In Bolivian specimens, Stat-Pak and Trypanosoma Detect tests were 87.5% and 90.7% sensitive, respectively; both showed 100% specificity. Sensitivity in Peruvian specimens was much lower: 26.6-33.0% (Stat-Pak) and 54.3-55.2% (Trypanosoma Detect); both had specificities > 98%. Even in Bolivian specimens, these sensitivities are inadequate for stand-alone screening. The low sensitivity in Peru may be related to parasite strain differences. Chagas disease rapid tests should be field tested in each geographic site before widespread implementation for screening. Copyrigh

Cellular Integrin α5β1 and Exosomal ADAM17 Mediate the Binding and Uptake of Exosomes Produced by Colorectal Carcinoma Cells

Approximately 25% of colorectal cancer (CRC) patients develop peritoneal metastasis, a condition associated with a bleak prognosis. The CRC peritoneal dissemination cascade involves the shedding of cancer cells from the primary tumor, their transport through the peritoneal cavity, their adhesion to the peritoneal mesothelial cells (PMCs) that line all peritoneal organs, and invasion of cancer cells through this mesothelial cell barrier and underlying stroma to establish new metastatic foci. Exosomes produced by cancer cells have been shown to influence many processes related to cancer progression and metastasis. In epithelial ovarian cancer these extracellular vesicles (EVs) have been shown to favor different steps of the peritoneal dissemination cascade by changing the functional phenotype of cancer cells and PMCs. Little is currently known, however, about the roles played by exosomes in the pathogenesis and peritoneal metastasis cascade of CRC and especially about the molecules that mediate their interaction and uptake by target PMCs and tumor cells. We isolated exosomes by size−exclusion chromatography from CRC cells and performed cell-adhesion assays to immobilized exosomes in the presence of blocking antibodies against surface proteins and measured the uptake of fluorescently-labelled exosomes. We report here that the interaction between integrin α5β1 on CRC cells (and PMCs) and its ligand ADAM17 on exosomes mediated the binding and uptake of CRC-derived exosomes. Furthermore, this process was negatively regulated by the expression of tetraspanin CD9 on exosomes

Clinical Outcomes of a Zika Virus Mother-Child Pair Cohort in Spain

BACKGROUND: Zika virus (ZIKV) infection has been associated with congenital microcephaly and other neurodevelopmental abnormalities. There is little published research on the effect of maternal ZIKV infection in a non-endemic European region. We aimed to describe the outcomes of pregnant travelers diagnosed as ZIKV-infected in Spain, and their exposed children. METHODS: This prospective observational cohort study of nine referral hospitals enrolled pregnant women (PW) who travelled to endemic areas during their pregnancy or the two previous months, or those whose sexual partners visited endemic areas in the previous 6 months. Infants of ZIKV-infected mothers were followed for about two years. RESULTS: ZIKV infection was diagnosed in 163 PW; 112 (70%) were asymptomatic and 24 (14.7%) were confirmed cases. Among 143 infants, 14 (9.8%) had adverse outcomes during follow-up; three had a congenital Zika syndrome (CZS), and 11 other potential Zika-related outcomes. The overall incidence of CZS was 2.1% (95%CI: 0.4-6.0%), but among infants born to ZIKV-confirmed mothers, this increased to 15.8% (95%CI: 3.4-39.6%). CONCLUSIONS: A nearly 10% overall risk of neurologic and hearing adverse outcomes was found in ZIKV-exposed children born to a ZIKV-infected traveler PW. Longer-term follow-up of these children is needed to assess whether there are any later-onset manifestations

Erratum to: 36th International Symposium on Intensive Care and Emergency Medicine

[This corrects the article DOI: 10.1186/s13054-016-1208-6.]

A922 Sequential measurement of 1 hour creatinine clearance (1-CRCL) in critically ill patients at risk of acute kidney injury (AKI)

Meeting abstrac

El ajuar de oro de la tumba fenicia del Obispo. Cádiz

Las recientes excavaciones efectuadas en el palacio episcopal, edificio anejo a la Catedral de la actual Cádiz, con motivo de su remodelación, sacaron a la luz un Unportante yacimiento arqueológico que presenta una ocupación continuada desde el siglo Wi a.c. hasta nuestros días. Uno de los hallazgos más importantes ha sido una tumba fenicia monumental, con cámara de sillares de piedra y probable superestructura arquitectónica, cuyos elementos verticales practicamente han desaparecido, pero que debió pertenecer al tipo de templo-tumba que podemos fechar hacia finales del siglo VI a.C. El ajuar había sido expoliado, pero todavía se pudo recuperar un anillo de oro y unas pequeñas laminillas del mismo metal que formaban parte de un rico tejido. En esta comunicación presentamos los primeros resultados del estudio de los objetos de oro que se analizaron en microsonda de Microscopio Electrónico de Barrido (MEB) y mediante PIXE.Peer reviewe

Purification, immobilization and stabilization of a highly enantioselective alcohol dehydrogenase from Thermus thermophilus HB27 cloned in E. coli

Funding This work has been supported by CAM (grants S0505/PPQ/0344) and MEC (grants BIO2004-02671 and CTQ2005-02420/PPQ). An institutional grant from Fundación Ramón Areces to CBMSO is also acknowledged. J.M. Bolívar and J. Rocha-Martín are holders of a PhD fellowship from CAM. Dr. Ángel Berenguer (Universidad de Alicante) is gratefully acknowledged for his kind help during the writing of this paper.This paper shows the purification and immobilization of a very interesting thermophilic alcohol dehydrogenase from Thermus thermophilus HB27 cloned in Escherichia coli. The purification was based on a first thermal treatment of the crude extract, that leaves the target enzyme in the supernatant, followed by the adsorption of most contaminant proteins in a IMAC column (the target protein did not adsorb on these columns due to the poorness of His residues). Final purification factor was around a 9-fold factor (no other protein bands were detected in SDS-PAGE gels) with an overall yield around 80%. Covalent immobilization of the enzyme on very different supports only permitted to improve the enzyme stability by a 5–10-fold factor, very similarly to the results obtained by the adsorption of the enzyme on polyethyleneimine coated supports. This enzyme adsorbed by ionic exchange maintained the activity unaltered during immobilization which was a very rapid process, and was more stable than the covalent preparations in the presence of organic solvents, and the enzyme was quite strongly adsorbed on the support. Therefore, it was proposed as a good option to prepare industrial biocatalysts of the enzyme. This preparation was utilized in the asymmetric reduction of acetophenone to produce (S)-(−)-1-phenylethanol, with an enantiomeric excess of more than 99%.Comunidad de MadridMinisterio de Educación y Ciencia (MEC)Fundación Ramón ArecesDepto. de Bioquímica y Biología MolecularFac. de Ciencias BiológicasFac. de Ciencias QuímicasTRUEpu