Aurora kinase B inhibition reduces the proliferation of metastatic melanoma cells and enhances the response to chemotherapy

Background: The poor response to chemotherapy and the brief response to vemurafenib in metastatic melanoma patients, make the identification of new therapeutic approaches an urgent need. Interestingly the increased expression and activity of the Aurora kinase B during melanoma progression suggests it as a promising therapeutic target. Methods: The efficacy of the Aurora B kinase inhibitor barasertib-HQPA was evaluated in BRAF mutated cells, sensitive and made resistant to vemurafenib after chronic exposure to the drug, and in BRAF wild type cells. The drug effectiveness has been evaluated as cell growth inhibition, cell cycle progression and cell migration. In addition, cellular effectors of drug resistance and response were investigated. Results: The characterization of the effectors responsible for the resistance to vemurafenib evidenced the increased expression of MITF or the activation of Erk1/2 and p-38 kinases in the newly established cell lines with a phenotype resistant to vemurafenib. The sensitivity of cells to barasertib-HQPA was irrespective of BRAF mutational status. Barasertib-HQPA induced the mitotic catastrophe, ultimately causing apoptosis and necrosis of cells, inhibited cell migration and strongly affected the glycolytic metabolism of cells inducing the release of lactate. In association i) with vemurafenib the gain in effectiveness was found only in BRAF(V600K) cells while ii) with nab-paclitaxel, the combination was more effective than each drug alone in all cells. Conclusions: These findings suggest barasertib as a new therapeutic agent and as enhancer of chemotherapy in metastatic melanoma treatment

Phytoremediation of soils contaminated with lead by arundo donax L

Collaborative project, FP7-KBBE-2011.3.1-02Arundo donax L. is a high yielded perennial energy crop. Its cultivation for bioenergy may generate land-use conflicts which might be avoided through the establishment of energy crops on marginal land. In this context, this work aims to study the potentiality of Arundo donax in Pb contaminated soils (450 and 900 mg Pb kg −1 , dry matter) under a low irrigation regime (475 mm). Results showed that biomass productivity of giant reed was negatively affected by the contamination. Increased higher lead content in the biomass was obtained with increased lead contamination of the soil. Highest accumulation of lead was observed in the roots and rhizomes. Although the lead removal percentages by giant reed accumulation represent 3.2‰ maximum, after two consecutive years, towards the Pb soil bioavailable fraction, the establishment of a vegetative cover represent an approach to attenuate and stabilize contaminated sites with additional revenue to owners.publishe

Barasertib: a novel approach for the treatment of metastatic melanoma

Background: Metastatic melanoma represents the most deadly form of skin cancer. The poor response to chemotherapy and the brief response to the anti-BRAF vemurafenib in selected population of patients, make the identification of new therapeutic approaches an urgent need. Our goal is the evaluation of the efficacy of barasertib, an aurora B kinase inhibitor impairing cytokinesis, in both mutated and non-mutated melanoma cell lines. Materials and methods: Panel of melanoma cells: BRAFV600E mutated cells (MBA72 and Hmel1), the same cell in which the resistance to vemurafenib was induced by chronic exposure to it (MBA72R and Hmel1R) and BRAF wt (HBL and LND1). Cells were characterized for vemurafenib and barasertib effectiveness on cell growth by MTT assay after 3 and 6 days of continuous exposure. Cell cycle was determined by flowcytometry and migration was evaluated by wound-healing assay. Results: Cells with BRAFV600E mutation are sensitive to vemurafenib conversely, those with BRAF wt and the resistant ones showed an IC50 of at least 10 folds higher. 3days-barasertib exposure strongly reduced cell growth (30-60% at 30 and 300nM, respectively) in all cell lines; when the drug was given together with vemurafenib, no gain in effectiveness was evident. Prolonged exposure to barasertib (6 days) showed a progressive increase of effectiveness particularly in cells BRAF wt. The analysis of cell death mechanisms involved in determining the effectiveness of barasertib and vemurafenib showed that the first drug induced both apoptosis and necrosis conversely, the latter mainly apoptosis. Cell cycle analysis demonstrated that barasertib induced an increase in cell size and in polyploidia, suggesting also the mitotic catastrophe as a further cell death mechanism. Moreover, the anti-metastatic behaviour of this agent has been evaluated in function of drug concentration and time exposure. Preliminary results showed a strong reduction of cell migration after drug exposure. Conclusions: The sensitivity of melanoma cells to barasertib is irrespective to BRAF mutational status; however, cells BRAF wt show an higher nuclear modification. In conclusion, our results suggest barasertib as a novel therapeutic approach in melanoma treatment irrespective of BRAFV600E mutation

HIV-infected patients show impaired cellular immune response to influenza vaccination compared to healthy subjects

Detailed data on cellular immune response to influenza vaccination in HIV-infected patients are lacking. We analyzed cellular (IL-2, IL-4, IL-5, IL-10, IL-12, IL-13, IFN-\u3b3, TNF-\u3b1, GM-CSF) and humoral (IgG and IgM) immune response in 81 HIV-infected and 30 HIV-negative subjects, before (T0) and 4 weeks (T1) after receiving a single dose of trivalent MF59-adjuvanted influenza vaccine. No difference in humoral response (IgG or IgM) was demonstrated between the two groups. While an increase in most cytokines from T0 to T1 was observed in HIV-uninfected subjects, cytokines production did not significantly increased in HIV-infected patients. Exploring Th1 response, higher CD8 cells count was significantly associated with lower post-vaccination IFN\u3b3 levels, while a higher CD4 cells count was associated with a greater response. Exploring Th2 response, higher HIV viral load was significantly associated with reduced post-vaccination IL-10 levels. In conclusion, in HIV-infected patients influenza vaccination could have good efficacy in sustaining humoral response but cellular response appeared impaired

Naturally occurring hepatitis C virus (HCV) NS3/4A protease inhibitor resistance-related mutations in HCV genotype 1-infected subjects in Italy

To assess the prevalence of hepatitis C virus (HCV) NS3/4A protease inhibitor (PI) resistance mutations in HCV genotype 1-infected PI-naive individuals in Italy