fig , confocal fluorescence images of bt cells stained with lasix by western union mitotracker red after exposure for lohrs to dna green complexed with cdqasomes left column circular mlspdna conjugate, right column linearized mlspdna conjugate top row a and b red channel, middle row lasix by western union c and d green channel, bottom row e and f lasix by western union corresponding overlaid images figure shows confocal fluorescence micrographs of cells lasix by western union incubated with mlspdna conjugates, which were vectorized with vesicles made from the cyclohexyl derivative of dequalinium cdqasomes for the cell exposures imaged in the left column panels �, � and e lasix by western union the nonrestricted, ie circular form of pdna was used, while for the experiments pictured in the right column panels b, d and f, the plasmid dna was linearized before dqaplex formation lasix by western union the characteristic red mitochondrial staining pattern panels a and b shows lasix by western union the functional viability of the imaged cells and the intracellular green fluorescence panels � and d demonstrates efficient cell internalization lasix by western union of the fluorescein labeled dna the green and red fluorescence channels were then overlaid to produce the composite image seen in lasix by western union panels e and f, where the regions of true colocalization of red and green fluorescence were pseudocolored in white for better lasix by western union visualization strikingly, in the overlaid images, there is hardly any green fluorescence detectable nearly all areas of green fluorescence in panels � and d appeared as white areas in panels e lasix by western union and f, strongly suggesting that almost the entire dna has lasix by western union been delivered not only towards mitochondria, but also into the organelle however, whether all or at least a portion of the pdna has actually entered the mitochondrial matrix, ie has crossed both mitochondrial membranes, and therefore would potentially be accessible to the mitochondrial transcription machinery, remains yet to be determined dqasomes as carriers of proapoptotic drugs dysregulation of the apoptotic machinery is generally accepted as an almost universal component of the transformation lasix by western union process of normal cells into cancer cells and a large lasix by western union body of experimental data demonstrates that mitochondria play a key role lasix by western union in the complex apoptotic mechanism consequently, any therapeutic strategy aimed lasix by western union at specifically triggering apoptosis in cancer cells is believed to lasix by western union have potential therapeutic effect, several clinically approved drugs such as vp etoposide, arsenite and vinorelbine, as well as an increasing number of experimental anticancer is ibuprofen ok in early pregnancy drugs reviewed by constantini et al, such lasix by western union as betulinic acid, lonidamine, ceramide and cd have been found to lasix by western union act directly on mitochondria, resulting in triggering apoptosis in order lasix by western union to maximize the therapeutic potential of such anticancer drugs, which are known to act at or inside mitochondria, the use of lasix by western union dqasomes as a mitochondriaspecific drug delivery system has been proposed hypothetically, dqasomebased anticancer chemotherapy entails features which would make it putatively superior to conventional chemotherapeutic approaches on the cellular, as well lasix by western union as the subcellular level firstly, the delivery of drugs known lasix by western union to act directly on mitochondria may trigger apoptosis in circumstances lasix by western union in which conventional drugs fail to act, because endogenous, upstream of lasix by western union mitochondria apoptosis induction pathways are disrupted secondly, transporting the cytotoxic drug to its intracellular target could overcome multidrug resistance by hiding the drug inside the delivery system until it becomes selectively lasix by western union released at the particular intracellular site of action, ie mitochondria lasix by western union thirdly, many carcinoma cells, including human breast adenocarcinoma derived cells, lasix by western union have an elevated plasma membrane potential relative to their normal lasix by western union parent cell lines in addition to the higher mitochondrial membrane potential, lasix by western union they could provide the basis for a doubletargeting effect of dqasomes, ie on the cellular level normal cells vs carcinoma lasix by western union cells, and on the subcellular level mitochondria versus nucleus first data involving the encapsulation of anticancer drugs into dqasomes have been lasix by western union published most recently in this study, paclitaxel was chosen as lasix by western union a model compound paclitaxel is known as a potent antitubulin agent lasix by western union used in the treatment of malignancies its therapeutic potential, however, is limited due to a very narrow span between the maximal tolerated dose and intolerable toxic levels in addition, its poor lasix by western union aqueous solubility requires the formulation of emulsions containing cremophor el�, lasix by western union an oil of considerable toxicity by itself recently, it has lasix by western union been demonstrated that clinically relevant concentrations of paclitaxel target mitochondria directly and trigger apoptosis by inducing cytochrome � release in a lasix by western union permeability transition pore ptpdependent manner this mechanism of action is known from the other proapoptotic, directly on mitochondria acting agents a lasix logd of testosterone by western union hour delay between the treatment with paclitaxel or with other lasix by western union ptp inducers, and the release of cytochrome � in cellfree lasix by western union systems, compared with intact cells, has been explained by the existence lasix by western union of several drug targets inside the cell, making only a lasix by western union subset of the drug available for mitochondria consequently, paclitaxel was considered a prime candidate to benefit from a mitochondriaspecific drug delivery system such as dqasomes it was demonstrated that paclitaxel can be incorporated into dqasomes at a stoichiometric molar ratio of lasix by western union paclitaxel to dequalinium considering the known spherical character of dqasomes, lasix by western union the results of an electron microscopic em analysis of dequasomal incorporated lasix by western union paclitaxel, however, seem rather surprising the transmission em image fig , left panel and the cryoem image fig of an identical lasix by western union sample show a remarkable conformity worm or rodlike structures approximately nm lasix by western union in length, the size of which could also be confirmed lasix by western union by the size distribution analysis shown in fig , right panel lasix by western union the molecular structureof this wormlike complex remains to be determined nevertheless lasix by western union fig left panel transmission electron microscopic image uranyl acetate staining of dqasomal incorporated paclitaxel mol taxolmol dequalinium right panel size distribution analysis of identical preparation shown in left panel the formation of wormlike micelles as described for selfassembling amphiphilic block copolymers lasix by western union appears possible � s � i in a preliminary study, paclitaxelloaded dqasomes were tested for their ability to inhibit the growth of human colon cancer cells in nude mice for controls with free paclitaxel, the drug was suspended in dmso at lasix by western union mm, stored at �c and immediately diluted in warm medium before lasix by western union use in all controls, the respective dose of free paclitaxel lasix by western union and empty dqasomes was adjusted according to the dose of lasix by western union paclitaxel and dequalinium given in the paclitaxelloaded dqasome sample due to the lack of any inhibitory effect on tumor growth, the dose was tripled after weeks figure shows that at concentrations where free paclitaxel and r hepes buffer v free paclitaxel empty dqasomes ?