Anti-inflammatory effects of tobramycin and a copper-tobramycin complex with superoxide dismutase-like activity.

BACKGROUND AND PURPOSE: Airway inflammation in cystic fibrosis (CF) patients is characterized by accumulations of neutrophils in the airway and T cells in bronchial tissue, with activation of platelets in the circulation. CF patients are routinely treated with systemic or inhaled tobramycin for airway infection with Pseudomonas aeruginosa. Clinical trials have indicated an anti-inflammatory effect of tobramycin beyond its bactericidal activity. Here, we investigate the anti-inflammatory properties of tobramycin in vitro and consider if these relate to the ability of tobramycin to bind copper, which is elevated in blood and sputum in CF. EXPERIMENTAL APPROACH: A copper-tobramycin complex was synthesized. The effect of tobramycin and copper-tobramycin on neutrophil activation and migration of T cells and neutrophils across human lung microvascular endothelial cells in response to thrombin-activated platelets were investigated in vitro. Tobramycin uptake was detected by immunocytochemistry. Intracellular reactive oxygen species were detected using the fluorescent indicator, 2',7'-dichlorofluorescein diacetate (DCFDA). Neutrophil superoxide, hydrogen peroxide and neutrophil elastase activity were measured using specific substrates. Copper was measured using atomic absorption spectroscopy. KEY RESULTS: Tobramycin and copper-tobramycin were taken up by endothelial cells via a heparan sulphate-dependent mechanism and significantly inhibited T-cell and neutrophil transendothelial migration respectively. Copper-tobramycin has intracellular and extracellular superoxide dismutase-like activity. Neutrophil elastase inhibition by α1-antitrypsin is enhanced in the presence of copper-tobramycin. Tobramycin and copper-tobramycin are equally effective anti-pseudomonal antibiotics. CONCLUSIONS AND IMPLICATIONS: Anti-inflammatory effects of tobramycin in vivo may relate to the spontaneous formation of a copper-tobramycin complex, implying that copper-tobramycin may be more effective therapy.

Preventing bacterial adhesion onto surfaces: the low-surface-energy approach.

Good-quality coatings prepared from poly(methylpropenoxyfluoroalkylsiloxane)s or poly(perfluoroacrylate)s are capable of inhibiting the bacterial colonisation of surfaces.

The surface properties of silicone elastomers Exposed to seawater.

The performances of some silicone elastomers as compliant coatings which are resistant to marine fouling have been assessed from a sea-water exposure trial covering three fouling seasons. Measurements of contact angles (polar and non-polar liquids, recently-advanced and recently-receded liquid drops and air bubbles) have been used to investigate the surface properties of materials and of coatings resistant to fouling after two years' exposure. The unmodified poly (dimethyIsiloxane) elastomer General Electric (GE) 21 was still resistant to marine settlement after three seasons and the poly(dimethyldiphenylsiloxane) GE655 only became fouled during the third season. No other unmodified material showed resistance to fouling beyond two seasons. The addition of a low-viscosity poly(dimethylsiloxane) oil to GE655 in a sufficient quantity (20 mass %) to cause blooming resulted in a material that remained free of fouling. Time-dependent behaviour by drops of all liquids on freshly prepared samples was observed in recently-advanced contact angles but not by recently-receded contact angles. With polar liquids, hard clear elastomers showed stepwise changes and also gave considerable contact-angle hysteresis effects. Immersion in water over a period of several weeks brought about a slow decrease in the hydrophobicity of all elastomers. GE21, after exposure in seawater for over two years, also showed a decrease as indicated by the contact angle of distilled water drops on its surface. The slow changes in the interfacial properties of silicones with polar liquids are attributed to rearrangements of polymer chains close to the surface, driven by the formation of hydrogen bonds between the solvent and oxygen atoms in the backbone. Penetration of the material by water gradually increases the surface energy and, sooner or later, the material becomes susceptible to fouling. For GE655, this may be delayed by incorporating with the formulation a relatively incompatible low-viscosity silicone oil.

Evaluation of the electrophilicity of DNA-binding pyrrolo[2,1-c][1,4]benzodiazepines by HPLC.

An HPLC assay is described that can be used to study the covalent bonding interaction of carbinolamine-containing pyrrolo[2,1-c][1,4]benzodiazepines with the model nucleophile thiophenol, in order to evaluate electrophilicity at the C-11-position. Preliminary experiments with anthramycin, tomaymycin and neothramycin show that their reaction with thiophenol follows second-order kinetics, but the ranking order of reactivity (neothramycin greater than tomaymycin greater than anthramycin), does not correlate with either in vitro cytotoxicity or in vivo antitumour activity. This suggests that other factors such as non-covalent DNA-interaction or drug transport play a more crucial role in biological activity than simple alkylating ability. This assay should, however, prove a useful tool in the study of structure-activity relationships for this series of compounds and provide "C-11-electrophilicity" parameters for use in Hansch analysis and related studies.