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Introduction Next Generation Sequencing (NGS) platforms have radically changed the field of genomics and are routinely applied to a variety of functional genomics problems. Although NGS has markedly accelerated multiple areas of genomics research it is a massively parallel process and generates unprecedented volumes of data which present challenges and opportunities for data management, storage, and, most importantly, analysis and interpretation. The Molecular Biodiversity Laboratory (MoBiLab), located in Bari at the CNR-IBBE, is a NGS research infrastructure, realized within the LifeWatch European infrastructure for Biodiversity. In MoBiLab, skills and advanced facilities for molecular and bioinformatics analyses are integrated to provide the scientific community with services and counselling for molecular biodiversity studies. MoBiLab has started the set up of a quality management system based on Good Laboratory Practices (GLPs) accompanied by a Laboratory Information Management System (L.I.M.S.) in order to assure the highest levels of reliability, reproducibility and traceability of the results. Methods GLPs are considered a quality system aimed at ensuring safety, reliability and reproducibility of the non-clinical tests on chemicals intended for the use on human, animal and environment. GLPs are mandatory in OECD countries for preclinical tests, but may be also considered as a reference for laboratory management systems outside their main scope, that can be referred to as “GLP-like” quality systems. The L.I.M.S. is a computer system capable of handling the Acquisition stages-Processing-Storage of all data generated by a laboratory and/or processes, minimizing the risk of errors and increasing the information security. Results The internal quality system of MoBiLab has been designed by taking into account the progressive evolution of Good Research Practices. The team has first outlined a description of the main process by means of a Supplier-Input-Process-Output-Customer-like flowchart which includes the person in charge for the activity and the related documented information (Standard Operation Procedures, SOP, and records). After having identified the SOPs, researchers were provided with a template and with the instructions to draft them. In parallel, management procedures were defined by the whole team, drafted, and supported by flowcharts and other quality tools. MoBiLab, in collaboration with the Italian company Eusoft, started to develop and optimize a L.I.M.S. platform for managing all the laboratory activities through a suite of integrated modules. The platform is structured starting from the SIPOC-like flowchart and the modules are developed and customized in agreement to the SOPs. A CAPA (Corrective Action/Preventive Action) system is also going to be implemented within the LIMS. Conclusions and Perspectives The implementation of some Lean tools in the production process of the MoBiLab will be the next task of the project.

We investigated the PCNA immunoreactivity in 35 specimens of malignant mesothelioma and 20 specimens of mesothelial hyperplasia in order to evaluate the usefulness of this parameter in differentiating between these two mesothelial proliferations, and to determine whether PCNA has any prognostic significance in mesotheliomas. Eleven of the 35 investigated malignant mesotheliomas displayed up to 25% of positive cells for PCNA expression. The remaining 24 specimens showed high percentages of positive cells ranging from 26% to 95%. All specimens of reactive hyperplasia had less than 25% of PCNA positive cells. The difference between malignant mesothelioma and mesothelial hyperplasia for PCNA immunoreactivity was statistically significant (p < 0.01). A positive relationship was also found between PCNA expression level and the overall survival of those affected by malignant mesothelioma (p = 0.0032). Our results suggest an important role for PCNA in differentiating diagnosis of mesothelial proliferations. It remains unclear whether PCNA expression truly correlates with the proliferation rate of the malignant mesotheliomas and with the overall survival of patients affected by this neoplasm.

Caveolae are plasma membrane subcompartments that have been implicated in signal transduction. In many cellular systems, caveolae are rich in signal transduction molecules such as G proteins and receptor-associated tyrosine kinases. An important structural component of the caveolae is caveolin. Recent evidence show that among the caveolin gene family, caveolin-3 is expressed in skeletal and cardiac muscle and caveolae are present in cardiac myocyte cells. Both the ANP receptor as well as the muscarinic receptor have been localized to the caveolae of cardiac myocytes in culture. These findings prompted us to conduct a further analysis of cardiac caveolae. In order to improve our understanding of the mechanisms of signal transduction regulation in cardiac myocytes, we isolated cardiac caveolae by discontinuous sucrose density gradient centrifugation from rat ventricles and rat neonatal cardiocytes. Our analysis of caveolar content demonstrates that heterotrimeric G proteins, p21ras and receptor-associated tyrosine kinases are concentrated within these structures. We also show that adrenergic stimulation induces an increase in the amount of diverse a- and b-subunits of G proteins, as well as p21ras, in both in vivo and in vitro experimental settings. Our data show that cardiac caveolae are an important site of signal transduction regulation. This finding suggests a potential role for these structures in physiological and pathological states. J. Cell. Biochem. 77:529-539, 2000. © 2000 Wiley-Liss, Inc.