Computational and experimental approaches assess the interactions between bovine \u3b2-lactoglobulin and synthetic compounds of pharmacological interest

Extending a previous investigation, the ability of binding to the model calycin \uf062-lactoglobulin (BLG) was evaluated both in silico and in vitro for several fluorine-containing (semi-)synthetic molecules of pharmacological and pharmaceutical interest (antibiotics, vastatins, steroid drugs). Simulation procedures included molecular docking according to a Montecarlo-simulated annealing protocol and molecular dynamics; heteronuclear NMR and denaturant gradient gel electrophoresis were the selected experimental techniques. For the tested drugs, ranking of the binding affinity was consistently assessed by computation and by experiment. The affinity for BLG increased in the sequence: 5-fluorosalycilic acid < dexamethasone << sulindac = norfloxacin < fluvastatin. The computed Ki for fluorosalycilate was in the order of 10-4 M; accordingly, in a molecular dynamics simulation the chemical diffused out of the BLG calyx in less than 2 ns, and no evidence of binding was found by NMR or electrophoresis. Conversely, the Ki for fluvastatin and norfloxacin were in the order of 10-7 and 10-6 M, similar to the affinity for BLG by natural ligands, such as retinoids and long-chain fatty acids. Moreover fluvastatin was found still bound to the protein after 5 ns of molecular dynamics simulation. Interaction of fluvastatin and norfloxacin with BLG was made evident by changes in chemical shift and dynamic parameters in the 19F NMR spectra and in effective urea concentration and cooperativity features in denaturant gradient gel electrophoresis. Such findings prove BLG may act as a drug carrier accepting in its cavity molecules of different bulk, rigidity and hydrophobicity

Characterization of canine mammary tumor initiating cells with stem cell potential

With the goal to indentifying the emerging concepts universal in tumor development from cancer stem cells, we have generated a dog cancer stem cell model system. Insight into the hierarchical organization of canine tumors as abnormal tissues that originates from, and are maintained by cancer sterm cells, should provide a mammalian tumor model system more similar to human than the rodent system

Isolation of canine mammary cells with stem cell properties and tumour initiating potential.

Recent data suggests that mammary carcinogenesis may be driven by cancer stem cells (CSCs) derived from mutated adult stem cells, which have acquired aberrant cell self-renewal or by progenitor cells that have acquired the capacity for cell self-renewal. Spontaneous mammary cancers in cats and dogs are important models for the understanding of human breast cancer and may represent alternative species model systems that can significantly contribute to the study of human oncogenesis. With the goal of identifying markers for isolating human breast CSCs, we have generated a canine model system to isolate and characterize normal and CSCs from dog mammary gland. Insight into the hierarchical organization of canine tumours may contribute to the development of universal concepts in oncogenesis by CSCs. Cells with stem cell properties were isolated from normal and tumoural canine breast tissue and propagated as mammospheres and tumourspheres in long-term non-adherent culture conditions. We showed that cells obtained from spheres that display self-renewing properties, have multi-lineage differentiation potential, could generate complex branched tubular structures in vitro and form tumours in NOD/SCID mice. We analysed these cells for the expression of human stem and CSC markers and are currently investigating the tumour-initiating properties of these cells and the hierarchical organization of normal and neoplastic canine mammary tissue

FarmaREL : an Italian pharmacovigilance project to monitor and evaluate adverse drug reactions in haematologic patients

Adverse drug reactions (ADRs) reduce patients' quality of life, increase mortality and morbidity, and have a negative economic impact on healthcare systems. Nevertheless, the importance of ADR reporting is often underestimated. The project "FarmaREL" has been developed to monitor and evaluate ADRs in haematological patients and to increase pharmacovigilance culture among haematology specialists. In 13 haematology units, based in Lombardy, Italy, a dedicated specialist with the task of encouraging ADRs reporting and sensitizing healthcare professionals to pharmacovigilance has been assigned. The ADRs occurring in haematological patients were collected electronically and then analysed with multiple logistic regression. Between January 2009 and December 2011, 887 reports were collected. The number of ADRs was higher in older adults (528; 59%), in male (490; 55%), and in non-Hodgkin lymphoma patients (343; 39%). Most reactions were severe (45% required or prolonged hospitalization), but in most cases, they were fully resolved at the time of reporting. According to Schumock and Thornton criteria, a percentage of ADRs as high as 7% was found to be preventable versus 2% according to reporter opinion. Patients' haematological diagnosis, not age or gender, resulted to be the variable that most influenced ADR, in particular severity and outcome. The employment of personnel specifically dedicated to pharmacovigilance is a successful strategy to improve the number and quality of ADR reports. "FarmaREL", the first programme of active pharmacovigilance in oncohaematologic patients, significantly contributed to reach the WHO "Gold Standard" for pharmacovigilance in Lombardy, Italy