No abstract

We investigated the influence of extracellular matrix on transport properties of mouse alveolar epithelial cell (AEC) monolayers (MAECM) and transdifferentiation of isolated mouse alveolar epithelial type II (AT2) cells into an alveolar epithelial type I (AT1) cell-like phenotype. Primary mouse AT2 cells plated on laminin 5-coated polycarbonate filters formed monolayers with transepithelial resistance (R(T)) and equivalent short-circuit current (I(EQ)) of 1.8 kOmega.cm(2) and 5.3 microA/cm(2), respectively, after 8 days in culture. Amiloride (10 microM), ouabain (0.1 mM), and pimozide (10 microM) decreased MAECM I(EQ) to 40%, 10%, and 65% of its initial value, respectively. Sequential addition of pimozide and amiloride, in either order, revealed that their inhibitory effects are additive, suggesting that cyclic nucleotide-gated channels contribute to amiloride-insensitive active ion transport across MAECM. Ussing chamber measurements of unidirectional ion fluxes across MAECM under short-circuit conditions indicated that net absorption of Na(+) in the apical-to-basolateral direction is comparable to net ion flux calculated from the observed short-circuit current: 0.38 and 0.33 microeq.cm(-2).h(-1), respectively. Between days 1 and 9 in culture, AEC demonstrated increased expression of aquaporin-5 protein, an AT1 cell marker, and decreased expression of pro-surfactant protein-C protein, an AT2 cell marker, consistent with transition to an AT1 cell-like phenotype. These results demonstrate that AT1 cell-like MAECM grown on laminin 5-coated polycarbonate filters exhibit active and passive transport properties that likely reflect the properties of intact mouse alveolar epithelium. This mouse in vitro model will enhance the study of AEC derived from mutant strains of mice and help define important structure-function correlations.

BCS ("Body Condition Score") is a method used to estimate body fat reserves and accumulated energy balance of cows. BCS heavily influences milk production, reproduction, and health of cows. Therefore, it is important to monitor BCS to achieve better animal response, but this is a time-consuming and subjective task performed visually by expert scorers. Several studies have tried to automate BCS of dairy cows by applying image analysis and machine learning techniques. This work analyzes these studies and proposes a system based on Convolutional Neural Networks (CNNs) to improve overall automatic BCS estimation, whose use might be extended beyond dairy production. The developed system has achieved good estimation results in comparison with other systems in the area. Overall accuracy of BCS estimations within 0.25 units of difference from true values was 78%, while overall accuracy within 0.50 units was 94%. Similarly, weighted precision and recall, which took into account imbalance BCS distribution in the built dataset, show similar values considering those error ranges. 1. uses images as the only information source (without external data such as weight, age, or lactation stage of the cow), 2. uses low-cost hardware resources, 3. gets real-time estimations,

The gas exchange surface of the lungs is lined by an epithelium consisting of alveolar type (AT) I and ATII cells. ATII cells function to produce surfactant, play a role in host defense and fluid and ion transport, and serve as progenitors. ATI cells are important for gas exchange and fluid and ion transport. Our understanding of the biology of these cells depends on the investigation of isolated cells. Here, we present methods for the isolation of mouse and rat ATII cells.