Antitumor effects of non-replicative herpes simplex vectors expressing antiangiogenic proteins and thymidine kinase on Lewis lung carcinoma establishment and growth.

There is growing evidence that combinations of antiangiogenic proteins with other antineoplastic treatments such as chemo- or radiotherapy and suicide genes-mediated tumor cytotoxicity lead to synergistic effects. In the present work, we tested the activity of two non-replicative herpes simplex virus (HSV)-1-based vectors, encoding human endostatin::angiostatin or endostatin::kringle5 fusion proteins in combination with HSV-1 thymidine kinase (TK) molecule, on endothelial cells (ECs) and Lewis lung carcinoma (LLC) cells. We observed a significant reduction of the in vitro growth, migration and tube formation by primary ECs upon direct infection with the two recombinant vectors or cultivation with conditioned media obtained from the vector-infected LLC cells. Moreover, direct cytotoxic effect of HSV-1 TK on both LLC and ECs was demonstrated. We then tested the vectors in vivo in two experimental settings, that is, LLC tumor growth or establishment, in C57BL/6 mice. The treatment of pre-established subcutaneous tumors with the recombinant vectors with ganciclovir (GCV) induced a significant reduction of tumor growth rate, while the in vitro infection of LLC cells with the antiangiogenic vectors before their implantation in mice flanks, either in presence or absence of GCV, completely abolished the tumor establishment.

Arsenic speciation and susceptibility to oxidative stress in the fanworm Sabella spallanzanii (Gmelin) (Annelida, Sabellidae) under naturally acidified conditions: An in situ transplant experiment in a Mediterranean CO2 vent system.

The fanworm Sabella spallanzanii (Gmelin, 1791) (Annelida, Sabellidae) is considered tolerant to several types of stressors but is generally absent from the CO2 vents. A peculiar characteristic of this species is the elevated content of arsenic in the gills, particularly dimethylarsinic acid (DMA), stored as an anti-predatory compound. In this study, modulation of trace metal levels, chemical speciation of arsenic and oxidative stress biomarkers were quantified in S. spallanzanii after a 30days transplant experiment into naturally acidified conditions in a Mediterranean vent system. No significant bioaccumulation of metals was observed in the thoracic tissues and branchial crowns after the translocation period, whereas variations occurred in the relative abundance of different arsenic compounds with the appearance of inorganic forms. The antioxidant system of translocated polychaetes exhibited a significant decrease of enzymatic activities of both catalase and glutathione peroxidases, and the impairment of the overall capability to neutralize hydroxyl radicals (OH). This highlighted an oxidative challenge primarily on the detoxification pathway of hydrogen peroxide. Overall low pH-elevated pCO2 may have detrimental effects on arsenic metabolism and oxidative status of S. spallanzanii, supporting the hypothesis of species-specific differences in vulnerability to ocean acidification.

Apoptosis in the OC-k3 immortalized cell line treated with different agents.

The aim of this study is to outline the mechanisms leading cochlear cells to die. We utilized an immortalized cell line (OC-k3 cells) derived from the organ of Corti of transgenic mice in order to perform in-depth biochemical studies with no limitations on sample size and number. We probed these cells with cisplatin and gentamicin, two drugs which display in vivo undesired ototoxic side-effects. We investigated cell viability, reactive oxygen species (ROS) production and glutathione (GSH) levels and tested the effects of different concentrations of cisplatin and gentamicin from 0 to 48 h. Results show that cells undergo a dose- and treatment-time-dependent apoptosis characterized by nuclear fragmentation, integrity of the cell membrane and mitochondria, and absence of DNA endonuclease activity. During the early part of treatment, ROS production increases and intracellular GSH decreases, probably due to the activation of protein kinase C alpha. Use of antioxidants such as N-acetylcysteine, GSH and vitamin C rescues cells from apoptosis almost completely. Overall, these data indicate that ROS generation might play a central role in inducing inner ear cell apoptosis and may have an additive role in the ageing process.