An environmental forensic approach for tropical estuaries based on metal bioaccumulation in tissues of Callinectes danae.

The blue crab Callinectes danae is distributed throughout the Atlantic coast and this study aimed to evaluate a environmental forensics approach that could be applied at tropical estuarine systems where this species is distributed, based on the metal concentrations in its tissues. For this purpose, blue crab samples were collected in 9 sites (distributed in 3 areas) along the Santos Estuarine System, state of São Paulo, Brazil. The concentrations of Al, Cd, Co, Cr, Cu, Fe, Hg, Mn, Ni, Pb and Zn were determined in gills, hepatopancreas and muscle tissues. Sediment samples were collected and analyzed in these same sites. A data distribution pattern was identified during both sampling periods (August and December 2011). In order to validate this model, a new sampling campaign was performed in March 2013 at the Santos Estuarine System and also at Ilha Grande (state of Rio de Janeiro). These data were added to the previous database (composed of the August and December 2011 samples) and a discriminant analysis was applied. The results confirmed an environmental fingerprint for the Santos Estuarine System.

Assessment of metal concentrations in muscles of the blue crab, Callinectes danae S., from the Santos Estuarine System.

This study determined the concentrations of eleven metals in the blue crab, Callinectes danae, from nine sites in the Santos Estuarine System of Sao Paulo State, Brazil. The results were compared to guidelines established in the United States, Europe and Brazil for the safety of human consumers. Muscles of blue crabs were removed by dissection and concentrations of Al, Cd, Co, Cr, Cu, Fe, Hg, Mn Ni, Pb and Zn were determined. In general, the concentrations of metals were low, and the crabs were regarded as safe for human consumption. Crabs from a single site (site 4) exceeded the guidelines established by the United States and Europe, but not Brazil, for Pb, with a mean tissue concentration of 1.725 µg g(-1). With the exception of Al, Fe and Ni, significant differences were noted between sites in the concentrations of each metal in crab tissue.

Survival of salivary gland cancer stem cells requires mTOR signaling.

Advanced salivary gland mucoepidermoid carcinoma (MEC) is a relentless cancer that exhibits resistance to conventional chemotherapy. As such, treatment for patients with advanced MEC is tipically radical surgery and radiotherapy. Facial disfigurement and poor quality of life are frequent treatment challenges, and many patients succumb to loco-regional recurrence and/or metastasis. We know that cancer stem-like cells (CSC) drive MEC tumorigenesis. The current study tests the hypothesis that MEC CSC are sensitive to therapeutic inhibition of mTOR. Here, we report a correlation between the long-term clinical outcomes of 17 MEC patients and the intratumoral expression of p-mTOR (p = 0.00294) and p-S6K1 (p = 0.00357). In vitro, we observed that MEC CSC exhibit constitutive activation of the mTOR signaling pathway (i.e., mTOR, AKT, and S6K1), unveiling a potential strategy for targeted ablation of these cells. Using a panel of inhibitors of the mTOR pathway, i.e., rapamycin and temsirolimus (mTOR inhibitors), buparlisib and LY294002 (AKT inhibitors), and PF4708671 (S6K1 inhibitor), we observed consistently dose-dependent decrease in the fraction of CSC, as well as inhibition of secondary sphere formation and self-renewal in three human MEC cell lines (UM-HMC-1,-3A,-3B). Notably, therapeutic inhibition of mTOR with rapamycin or temsirolimus induced preferential apoptosis of CSC, when compared to bulk tumor cells. In contrast, conventional chemotherapeutic drugs (cisplatin, paclitaxel) induced preferential apoptosis of bulk tumor cells and accumulation of CSC. In vivo, therapeutic inhibition of mTOR with temsirolimus caused ablation of CSC and downregulation of Bmi-1 expression (major inducer of stem cell self-renewal) in MEC xenografts. Transplantation of MEC cells genetically silenced for mTOR into immunodeficient mice corroborated the results obtained with temsirolimus. Collectively, these data demonstrated that mTOR signaling is required for CSC survival, and unveiled the therapeutic potential of targeting the mTOR pathway for elimination of highly tumorigenic cancer stem-like cells in salivary gland mucoepidermoid carcinoma.