29 research outputs found
Enhanced mass spectrometry-based proteomics can be used to distinguish differences in protein compositions of acellular emphysematous versus normal lungs.
Residual detergent detection method for nondestructive cytocompatibility evaluation of decellularized whole lung scaffolds.
The development of reliable tissue engineering methods using decellularized cadaveric or donor lungs could potentially provide a new source of lung tissue. The vast majority of current lung decellularization protocols are detergent based and incompletely removed residual detergents may have a deleterious impact on subsequent scaffold recellularization. Detergent removal and quality control measures that rigorously and reliably confirm removal, ideally utilizing nondestructive methods, are thus critical for generating optimal acellular scaffolds suitable for potential clinical translation. Using a modified and optimized version of a methylene blue-based detergent assay, we developed a straightforward, noninvasive method for easily and reliably detecting two of the most commonly utilized anionic detergents, sodium deoxycholate (SDC) and sodium dodecyl sulfate (SDS), in lung decellularization effluents. In parallel studies, we sought to determine the threshold of detergent concentration that was cytotoxic using four different representative human cell types utilized in the study of lung recellularization: human bronchial epithelial cells, human pulmonary vascular endothelial cells (CBF12), human lung fibroblasts, and human mesenchymal stem cells. Notably, different cells have varying thresholds for either SDC or SDS-based detergent-induced cytotoxicity. These studies demonstrate the importance of reliably removing residual detergents and argue that multiple cell lines should be tested in cytocompatibility-based assessments of acellular scaffolds. The detergent detection assay presented here is a useful nondestructive tool for assessing detergent removal in potential decellularization schemes or for use as a potential endpoint in future clinical schemes, generating acellular lungs using anionic detergent-based decellularization protocols
Decellularized wild type and alpha-galactosyltransferase knockout pig lungs stimulate M2 phenotype in human monocyte/macrophages.
Preclinical validation and imaging of Wnt-induced repair in human 3D lung tissue cultures.
Chronic obstructive pulmonary disease (COPD) is characterised by a progressive loss of lung tissue. Inducing repair processes within the adult diseased lung is of major interest and Wnt/β-catenin signalling represents a promising target for lung repair. However, the translation of novel therapeutic targets from model systems into clinical use remains a major challenge. We generated murine and patient-derived three-dimensional (3D) ex vivo lung tissue cultures (LTCs), which closely mimic the 3D lung microenvironment in vivo. Using two well-known glycogen synthase kinase-3β inhibitors, lithium chloride (LiCl) and CHIR 99021 (CT), we determined Wnt/β-catenin-driven lung repair processes in high spatiotemporal resolution using quantitative PCR, Western blotting, ELISA, (immuno)histological assessment, and four-dimensional confocal live tissue imaging. Viable 3D-LTCs exhibited preserved lung structure and function for up to 5 days. We demonstrate successful Wnt/β-catenin signal activation in murine and patient-derived 3D-LTCs from COPD patients. Wnt/β-catenin signalling led to increased alveolar epithelial cell marker expression, decreased matrix metalloproteinase-12 expression, as well as altered macrophage activity and elastin remodelling. Importantly, induction of surfactant protein C significantly correlated with disease stage (percent predicted forced expiratory volume in 1 s) in patient-derived 3D-LTCs. Patient-derived 3D-LTCs represent a valuable tool to analyse potential targets and drugs for lung repair. Enhanced Wnt/β-catenin signalling attenuated pathological features of patient-derived COPD 3D-LTCs
EFFECT OF CALIBRATION PROCEDURES ON ACCURACY AND PRECISION OF AUTOMATED KJELDAHL NITROGEN ANALYSIS IN SOME FORMULATED FOODS
An introductory tutorial on JastAdd attribute grammars
The final publication will be available at www.springerlink.com Abstract. JastAdd is an open-source system for generating compilers and other language-based tools. Its declarative specification language is based on reference attribute grammars and object-orientation. This allows tools to be implemented as composable extensible modules, as exemplified by JastAddJ, a complete extensible Java compiler. This tutorial gives an introduction to JastAdd and its core attribute grammar mechanisms, and how to use them when solving key problems in building language-based tools. A simple state machine language is used as a running example, showing the essence of name analysis, adding graphs to the abstract syntax tree, and computing circular properties like reachability. Exercises are included, and code for the examples is available online