No abstract

Large-pore channels, including those formed by connexin, pannexin, innexin proteins, are part of a broad family of plasma membrane channels found in vertebrates and invertebrates, which share topology features. Despite their relevance in parasitic diseases such as Chagas and malaria, it was unknown whether these large-pore channels are present in unicellular organisms. We identified 14 putative proteins in Trypanosomatidae parasites as presumptive homologs of innexin proteins. All proteins possess the canonical motif of the innexin family, a pentapeptide YYQWV, and 10 of them share a classical membrane topology of large-pore channels. A sequence similarity network analysis confirmed their closeness to innexin proteins. A bioinformatic model showed that a homolog of Trypanosoma cruzi (T. cruzi) could presumptively form a stable octamer channel with a highly positive electrostatic potential in the internal cavities and extracellular entrance due to the notable predominance of residues such as Arg or Lys. In vitro dye uptake assays showed that divalent cations-free solution increases YO-PRO-1 uptake and hyperosmotic stress increases DAPI uptake in epimastigotes of T. cruzi. Those effects were sensitive to probenecid. Furthermore, probenecid reduced the proliferation and transformation of T. cruzi. Moreover, probenecid or carbenoxolone increased the parasite sensitivity to antiparasitic drugs commonly used in therapy against Chagas. Our study suggests the existence of innexin homologs in unicellular organisms, which could be protein subunits of new large-pore channels in unicellular organisms.

La interacción entre la superficie y subsuperficie de una cuenca se puede estudiar utilizando modelos físicos, pero debido a su compleja geometría los procesos de calibración y validación requieren altos tiempos computacionales. Para ello, se propone una alternativa que permita reducir los tiempos de calibración del modelo, mediante la simplificación de la geometría de la cuenca y empleando el concepto del rectángulo equivalente. El rectángulo equivalente es usado para definir los parámetros del suelo de la cuenca del río Lerma, a través de un análisis de sensibilidad, que posteriormente son empleados dentro de la calibración. El modelo simplificado identifica con una gran precisión los rangos y valores iniciales de los parámetros utilizados para los procesos de calibración y además reduce significativamente los tiempos computacionales. Por lo tanto, el concepto de rectángulo equivalente ofrece una alternativa para acelerar la calibración de modelos acoplados superficie-subsuperficie.