Simplified cell culture method for the diagnosis of atypical primary ciliary dyskinesia.

BACKGROUND: The diagnosis of primary ciliary dyskinesia (PCD) can be challenging, and it may be particularly difficult to distinguish primary ciliary disease from the secondary changes after infections. OBJECTIVES: The purpose of the study was to evaluate if nasal epithelial cells, obtained with nasal brushing instead of a biopsy, could be used in a culture system for the diagnosis of PCD in difficult cases. METHODS AND MAIN RESULTS: Ciliary motion analysis (CMA) and transmission electron microscopy (TEM) were performed on 59 subjects with persistent or recurrent pneumonia. These investigations allowed the diagnosis of PCD in 13 (22%) patients while the defect of the cilia was considered secondary to infections in 37 (63%) subjects. In the remaining nine (15%) patients the diagnostic evaluation with CMA and TEM remained inconclusive. Ciliogenesis in culture allowed the diagnosis of PCD in four of these patients, it was indicative of a secondary defect in two subjects, and it was not helpful in the remaining three patients. CONCLUSIONS: Culture of cells obtained with brushing of the nasal turbinate is not a perfect test, nevertheless it may offer diagnostic help in doubtful cases of PCD.

HEMET: mathematical model of biochemical pathways for simulation and prediction of HEpatocyte METabolism.

Many computer studies and models have been developed in order to simulate cell biochemical pathways. The difficulty of integrating all the biochemical reactions that occur in a cell in a single model is the main reason for the poor results in the prediction and simulation of cell behaviour under different chemical and physical stimuli. In this paper we have translated biochemical reactions into differential equations for the development of modular model of metabolism of a hepatocyte cultured in static and standard conditions (in a plastic multiwell placed in an incubator at 37 degrees C with 5% of CO(2)). Using biochemical equations and energetic considerations a set of non-linear differential equations has been derived and implemented in Simulink. This set of equations mimics some of the principal metabolic pathways of biomolecules present in the culture medium. The software platform developed is subdivided into separate modules, each one describing a different metabolic pathway; they constitute a library which can be used for developing new modules and models to project, predict and validate cell behaviour in vitro.

Mandibular distraction osteogenesis: a systematic review of stability and the effects on hard and soft tissues.

Mandibular distraction osteogenesis (MDO) has been widely adopted in modern maxillofacial surgery due to its less invasive approach and the consistent aesthetic and functional improvements obtained. The aim of the present systematic review was to analyze the available evidence on the skeletal and soft tissue effects of MDO. The medical literature was searched to identify all peer-reviewed papers meeting the selection criteria for the final review process. A three-point grading system was used to rate the methodological quality of the selected papers. The PICO approach was used to extract data from the selected papers. The search strategy yielded eight relevant publications. The quality of the collected evidence was low to moderate. Vertical and sagittal skeletal dimensions increased significantly, by a mean of 5-10mm (P<0.05). Regarding the sagittal positioning of the lips and surrounding structures, a 90% correspondence between skeletal and soft tissue cephalometric points was observed. Significant skeletal relapse was reported, however it did not worsen the results of treatment significantly.

A new library of HEMET model: Insulin effects on hepatic metabolism.

Prediction and simulation of cell culture behaviour, under different chemical and physical stimuli by a mathematical model, represent an innovative way to create a virtual cell laboratory, where it is possible to perform and optimize experimental protocol, saving time and money. In silico experiments permit to reproduce pathological and physiological situations and make toxicological tests. In this paper we introduce a new library of HEMET (HEpatocyte METabolism) software that allows the insulin effects on hepatic metabolism to be simulated. This new set of nonlinear differential equations, derived from biochemical reactions which involve this pancreatic hormone, allows the catabolites concentration in hepatic cell culture after insulin infusion to be predicted. The validation procedures were carried out using data obtained from specifically designed cell experiments and from literature. A user friendly interface allows to easily change model parameters, rate constants and inputs simulating a wide range of physiological and pathological scenarios.

Rapid-prototyped and salt-leached PLGA scaffolds condition cell morpho-functional behavior.

Three dimensional scaffolds microfabricated using pressure-assisted microsyringe (PAM) with controlled geometry and porous membranes obtained using salt leaching were both tested with three different cell types to identify an optimal microstructural architecture for tissue engineering. MG63 (osteoblast-like cells) were used as models of mesenchymal bone tissue and human endothelial cells and NCTC2544 (keratinocytes) represented two epithelial tissues. Both porosity and stiffness of PLGA structures were measured, and cell morphology and cytoskeletal organization analyzed using SEM and actin labeling. The results show that overall the PAM scaffolds, which have a repeated and regular microstructure, are more biocompatible than the random pore salt-leached membranes, and that surface morphology as well as substrate stiffness modulates cell behavior.