³¹P NMR- and high-resolution ¹H MAS NMR studies of intact cancer and bacterial cells : an experimental investigaton of the effects induced by the Ca2+-ionophore A23187, daunorubicin and ALA-PDT
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NMR studies on cells have shown alterations in metabolic processes by the induction of apoptosis. The main purpose of the present study was to use NMR spectroscopic techniques to further elucidate metabolic and physiological processes involved in the cytotoxic response in leukemic cells. The metabolic processes of bacterial inactivation were also to be investigated. The following approaches were employed: 1. A perfusion system and a method for alginate immobilization of cells were developed for long-term studies (hours and days). The methods were subsequently used to perform 31P NMR spectroscopic studies on CLL cells under treatment by both PMA and A23187 in order to study processes of both activation and inactivation of the cells. Effects of the divalent cations Ca2+, Sr2+, and Ba2+ on viability, proliferation, and intracellular free calcium concentration were studied in Jurkat cells to further explore the applicability of Sr2+ in alginate-immobilization of cells. 2. The MAS technique was employed to obtain high-resolution 1H NMR spectra of the leukemic cells Jurkat, and 1D, 2D, and diffusion weighted NMR spectroscopy were used to study the cells after induction of apoptosis by DNR. 3. 1D time-series and 2D 1H MAS NMR spectroscopy were used to study the early metabolic events in ALA-PDT treated Jurkat cells. A dose-response 1H MAS NMR study was performed on ALA-PDT treated P. acnes to further study the effects of ALA-PDT.