Circulating miR-146a as a possible candidate biomarker in the indeterminate phase of Chagas disease

BACKGROUND: Chagas disease is considered important and presents intense inflammatory and fibrotic processes induced by the perpetuation of the parasite in the affected tissues and organs. Therefore, it is necessary to inquire about the host defense and attack mechanisms to have a more detailed knowledge about Chagas disease. Micro-RNAs are found in blood, tissues and extracellular vesicles. These small regulators of gene expression are involved in physiological and pathological processes in both mammals and parasites. Several microRNAs have deregulated expression in chagasic heart disease, although little is known about their extracellular expression. Our main objective was to evaluate the involvement of miR-21, miR-146a and miR-155 in several samples from mice infected with the TcI Ninoa strain from the acute and indeterminate phases. We also explored a potential functional association of the selected microRNAs using STRING software. This software identified 23 pathways associated with Trypanosoma cruzi infection. In addition, eleven genes were identified through bioinformatics analysis, and we found that SMAD family member 5 was downregulated in both phases. This gene serves as a mediator in the TGF-ß signaling pathway. Thus, forty female mice of the CD1 strain were distributed into 4 groups and the expression levels of miR-21, miR-146a and miR-155 were measured in samples of heart tissue, total plasma and plasma extracellular vesicles by quantitative real-time polymerase chain reaction. RESULTS: Overexpression of miR-21, miR-146a and miR-155 was observed in heart and plasma in both phases. Moreover, in extracellular vesicles miR-21 and miR-146a were also overexpressed in the acute phase, whereas in the indeterminate chronic phase we found only miR-146a up-regulated. CONCLUSIONS: The expression of inflammatory microRNAs miR-21, miR-146a and miR-155 were up-regulated in each of the samples from acutely and chronically infected mice. The relevant finding was that miR-146a was up-regulated in each sample in both phases; therefore, this miRNA could be a possible candidate biomarker in Chagas disease.

Effect of fresh frozen plasma on the in vitro activation of U937 monocytes: a potential role for the age of blood donors and their underlying cytokine profile

BACKGROUND: Fresh frozen plasma (FFP) administration may increase the risk of nosocomial infections in parallel with the development of immune modulation. This could be driven by soluble mediators, possibly influencing the in vitro activation of human U937 monocyte cells, in a manner dependent on the age of the donors. METHODS: FFP donors were stratified into groups of 19-30 years, 31-40 years or 41-50 years, and U937 cells were cultured with FFP (alone or plus lipopolysaccharide-LPS) for 24 h. Both in FFP and supernatants, TNF, IL-1ß, IL-6, and IL-10 levels were measured by ELISA. Additionally, CD11B, TLR2, and CASP3 gene expression were measured by qtPCR in U937 cells. Total phagocytic activity was also assayed. RESULTS: Elevated IL-10, but low TNF and IL-1ß levels were measured in FFP from individuals aged 19-40 years, whereas in individuals aged 41-50 years FFP were characterized by equalized TNF and IL-10 levels. Elevated IL-6 levels were found in all FFP samples, especially in those from the oldest individuals. FFP stimulation was associated with striking modifications in cytokine production in an age-dependent way. Exposure to FFP attenuates the response to LPS. TLR2 and CD11B expression were enhanced regardless of the age of plasma donors, although CASP3 expression was increased only when FFP from individuals aged 19-40 years were tested. Phagocytosis decreased after exposure to FFP regardless of donor age. CONCLUSION: Our results suggest that soluble mediators in FFP may modulate the functioning of monocytes. Interestingly, this effect appears to be partially influenced by the age of donors.

In vivo assessment of hemocompatibility of a ventricular assist device in healthy swine / Evaluación de la hemocompatibilidad in vivo (cerdos sanos) de un dispositivo de asistencia ventricular

Objective: To assess the hemocompatible performance of a novel implantable pneumatic ventricular assist device (VAD, Innovamédica, México) in healthy swine. The aim of this pilot study was first, to determine if short-term VAD implantation elicited a remarkable inflammatory response above that expected from surgical trauma; and second, to assess if heparinized or passivated VAD coatings, in combination with systemic anticoagulant or antiaggregant therapies, modified the VAD's hemocompatible performance. Methods: Hemodynamic, physiologic, inflammatory and histological parameters were measured in 27 pigs receiving VAD support for six hours, testing combinations of heparinized or passivated VAD coatings and systemic anticoagulant/ antiaggregant therapies. Mean concentrations of interleukin-1 β (IL-16), interleukin-6 (IL-6), C-reactive protein (CRP), or thrombin-antithrombin III (TAT) complexes (coagulation indicator) were measured from blood. ANOVA statistics were employed. Results: No substantial increases in mean IL-1β, IL-6, CRP, or TAT were obtained during VAD support. Hemodynamic and physiologic parameters were normal. We found no evidence of thromboembolisms or micro-infarctions in heart and lung samples. No major coaguli/deposits were found in VAD compartments. Overall, no remarkable differences in measurements were found using heparinized, passivated, or uncoated VAD, or with systemic anticoagulation, antiaggregant therapy, or no treatment. Conclusions: Our findings demonstrate, firstly, that during the time-period tested, the VAD elicited negligible inflammation above the effects of surgical trauma; and secondly, that little coagulation was observed upon VAD support in any of the cases tested. Contemplating further validation studies, our data indicate that the Innovamédica VAD is a highly hemocompatible system.

Objetivo: Evaluar la hemocompatibilidad de un nuevo dispositivo de asistencia ventricular (DAV, Innovamédica, México) neumático e implantable, en cerdos sanos. En este estudio piloto se propuso determinar primero, si la implantación a corto plazo del DAV suscitaría una respuesta inflamatoria por encima de aquella esperada tras trauma quirúrgico; segundo, evaluar si recubrimientos heparinizados o pasivos del DAV, en combinación con tratamientos sistémicos anticoagulantes o antiplaquetarios, modificarían la hemocompatibilidad del DAV. Métodos: Se midieron parámetros hemodinámicos, fisiológicos, inflamatorios e histológicos en 27 cerdos recibiendo soporte del DAV durante seis horas, evaluando combinaciones de recubrimientos heparinizados y pasivos del DAV, y terapias sistémicas anticoagulantes / antiplaquetarias. Se obtuvieron, a partir de sangre, las concentraciones promedio de interleucina-1 (IL-1β), interleucina-6 (IL-6), proteína C reactiva (PCR) y los complejos trombina-antitrombina III (TAT) (índice de coagulación). Se emplearon análisis estadísticos ANOVA. Resultados: No se observaron incrementos importantes en los niveles promedio de IL-1β, IL-6, PCR, o TAT durante soporte del DAV. Los parámetros hemodinámicos y fisiológicos fueron normales. No existió evidencia alguna de trom-boembolias o micro-infartos en muestras de miocardio y pulmón. No se encontraron coágulos o depósitos mayores en compartimentos del DAV. En general, no se apreciaron diferencias notables de mediciones utilizando dispositivos con recubrimiento heparinizado, pasivo o sin recubrimiento, en conjunto con terapia sistémica anticoagulante, antiplaquetaria o sin ella. Conclusiones: Nuestros hallazgos demuestran, primero, que durante el periodo de medición experimental, el DAV suscitó una respuesta inflamatoria mínima por encima de los efectos de trauma quirúrgico, y; segundo, en todos los casos evaluados, se observaron escasos o inexistentes efectos de coagulación durante soporte ventricular. Contemplando estudios adicionales de validación, nuestros datos indican que el DAV Innovamédica es un sistema altamente hemocompatible.