Examinando por Autor "Rodriguez, F."

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    Effect of a lung rest strategy during ECMO in a porcine acute lung injury model
    (SpringerOpen, 2015-10) Araos, J.; Cruces, P.; Tapia, P.; Alegria, L.; García, P.; Salomon, T.; Rodriguez, F.; Amthauer, M.; Castro, G.; Erranz, B.; Soto, D.; Carreño, P.; Medina, T.; Damiani, F.; Bugedo, G.; Bruhn, A.
    Introduction ECMO is used to treat patients who develop refractory hypoxemia and to provide a more protective ventilation. Several guidelines recommend “lung rest” strategies based on variable ventilatory parameters. However, there is limited evidence to support this strategy. Objectives To compare the effect of a lung rest strategy based on near-apneic ventilation (Vt 1-2 ml/kg, PEEP 10, respiratory rate-RR 5 min) versus conventional (Vt 10ml/kg, PEEP 5, RR 20/min), and standard protective ventilation (Vt 6ml/kg, PEEP 10, RR 20/min). Methods Twenty-four domestic pigs (26-36 kg) were anesthetized, mechanically ventilated (Vt 10 ml/kg, PEEP 5, O2 1.0) and invasively monitored. Six animals were used as Sham. In the other 18 lung injury was induced by saline lavages (30 ml/kg per lavage) performed repeatedly in both supine and prone position until PaO2/FiO2 dropped below 250. They were then subjected to a 2-hour injurious ventilation with PCV, PEEP = 0, Pinsp = 40 cmH2O, RR = 10/min, I:E = 1:1, one hour in prone and the other in supine. After completing lung injury (time 0) animals were connected to a saline primed-MEDOS Hilite ECMO circuit by inserting a AVALON 23F double-lumen cannula through the external jugular vein. Blood flow was set at 60-70% of cardiac output. Animals were randomized into one of the three groups and ventilated according to randomization for the following 24 hours. Respiratory and hemodynamic data were collected at times 0, 3, 6, 12, 18 and 24h. After euthanizing animals at time 24h, tissue samples were extracted from the lungs and injury evaluated and scored by light microscopy. Total lung water content was estimated by the wet-dry weight ratio. Results PaO2 decreased significantly in all groups after injury, but was progressively restored after ECMO start, despite the study group. Mean arterial pressure remained within normal limits throughout the study period, whereas MPAP increased significantly after injury but reached values close to SHAM soon after ECMO initiation. Lung wet-dry weight ratio and histological injury score increased significantly in all study groups compared to SHAM. Although non-significant, there was a trend towards a better histological injury score when Vt was reduced. Conclusions In this preliminary analysis, we found no clear advantage of reducing Vt when applying ECMO to support a double-hit animal model of ARDS in regard to resolution of lung edema or gas exchange. However, further work is required to determine if the non-significant reduction in lung injury observed in the near-apneic strategy may be relevant in providing further protection to the injured lungs supported by ECMO. © 2015 Araos et al.
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    Ítem
    Single and combined effect of retinoic acid and rapamycin modulate the generation, activity and homing potential of induced human regulatory T cells
    (Public Library of Science, 2017-07) Candia, E.; Reyes, P.; Covian, C.; Rodriguez, F.; Wainstein, N.; Morales, J.; Mosso, C.; Rosemblatt, M.; Fierro, J.A.
    Adoptive transfer of CD4+CD25+FOXP3+ regulatory T cells (Treg cells) has been successfully utilized to treat graft versus host disease and represents a promising strategy for the treatment of autoimmune diseases and transplant rejection. The aim of this study was to evaluate the effects of all-trans retinoic acid (atRA) and rapamycin (RAPA) on the number, phenotype, homing markers expression, DNA methylation, and function of induced human Treg cells in short-term cultures. Naive T cells were polyclonally stimulated and cultured for five days in the presence of different combinations of IL-2, TGF-β1, atRA and RAPA. The resulting cells were characterized by the expression of FOXP3, activation, surface and homing markers. Methylation of the Conserved Non-coding Sequence 2 was also evaluated. Functional comparison of the different culture conditions was performed by suppression assays in vitro. Culturing naive human T cells with IL-2/TGFβ1 resulted in the generation of 54.2% of Treg cells (CD4+CD25+FOXP3+) whereas the addition of 100 nM atRA increased the yield of Treg cells to 66% (p = 0.0088). The addition of RAPA did not increase the number of Treg cells in any of these settings. Treg cells generated in the presence of atRA had an increased expression of the β7 integrin to nearly 100% of the generated Treg cells, while RAPA treated cells showed enhanced expression of CXCR4. The differential expression of homing molecules highlights the possibility of inducing Treg cells with differential organ-specific homing properties. Neither atRA nor RAPA had an effect on the highly methylated CNS2 sites, supporting reports that their contribution to the lineage stability of Treg cells is not mediated by methylation changes in this locus. Treg cells generated in the presence of RAPA show the most potent suppression effect on the proliferation of effector cells.