Bromine indirubin FLIM/PLIM sensors to measure oxygen in normoxic and hypoxic PDT conditions.

BACKGROUND: The induction of phototoxicity during photodynamic therapy (PDT) is dependent on oxygen availability. For this reason, the development of sensors to measure oxygen and oxygen consumption is extremely important. APPROACH: In this project we have used Fluorescence Lifetime imaging (FLIM) and Phosphorescence Lifetime Imaging/ delayed Fluorescence Lifetime Imaging (PLIM/dFLIM) to investigate the ability of bromine indirubin derivatives as oxygen sensors. RESULTS: The oxygen sensitivity of bromine indirubins was detected through PLIM/dFLIM. Moreover, we have observed, by measuring nicotinamide adenine dinucleotide (NADH) FLIM, that bromine indirubin has a significant impact on cellular metabolism by shifting the SCC-4 Cells metabolism from oxidative phosphorylation (OXPHOS) to glycolysis. CONCLUSIONS: In conclusion, this study successfully achieves its goals and provides important insights into the use of indirubin as a potential oxygen consumption sensor with the capability to identify and differentiate between normoxic and hypoxic regions within the cells.

Pb2+ complexes of small-cavity azamacrocyclic ligands: thermodynamic and kinetic studies.

The synthesis, acid-base behavior and Pb2+ coordination chemistry of the new aza-scorpiand like ligand 5-[2-(N-2-fluorenyl)ethylamino]-2,5,8-triaza[9]-2,6-pyridinophane (L1) have been studied by potentiometry, NMR and spectrofluorimetric titrations, and the results are compared with those obtained for the related compounds L2, lacking the fluorenyl group, and L3, the macrocycle lacking the pendant arm. The crystal structures obtained for complexes [PbL1][PbL1Cl](NO3)Cl2·4H2O (1) and [PbL3](ClO4)2 (2) reveal that the metal ion is located over the plane defined by the nitrogen atoms of the macrocyclic core due to its inability to accommodate the large Pb2+ ion in the macrocyclic cavity. For L1, the secondary amino group of the pendant arm is implicated in the coordination of the metal ion, although the stereoactive lone pair of Pb2+ prevents the closed conformation associated with the coordination of metal ions in aza-scorpiand derivatives. The kinetics of the acid-promoted dissociation of the ligand from the Pb2+ complexes with the three ligands have been studied using stopped-flow with simultaneous absorbance and fluorescence detection. The results indicate that in spite of their similarity, the dissociation of the metal ion occurs with very different rates in the three complexes. During the course of the kinetic studies evidence was obtained for the occurrence of a photochemical process that leads to ligand degradation with the unexpected elimination of one CH2CH2 fragment from the macrocyclic core.