fig , confocal fluorescence images of bt cells stained with caffeine in fuze mitotracker red after exposure for lohrs to dna green complexed caffeine in fuze with cdqasomes left column circular mlspdna conjugate, right column linearized mlspdna conjugate top row a and b red channel, middle row c and d green channel, bottom row e and f corresponding overlaid images figure shows confocal fluorescence micrographs of cells incubated with mlspdna conjugates, which were vectorized with vesicles made from the cyclohexyl derivative of dequalinium cdqasomes for the cell exposures caffeine in fuze imaged in the left column panels �, � and e caffeine in fuze the nonrestricted, ie circular form of pdna was used, while for caffeine in fuze the experiments pictured in the right column panels b, d caffeine in fuze and f, the plasmid dna was linearized before dqaplex formation the characteristic red mitochondrial staining pattern panels a and b shows the functional viability of the imaged cells and the intracellular green fluorescence panels � and d demonstrates efficient cell internalization caffeine in fuze of the fluorescein labeled dna the green and red fluorescence channels were then overlaid to produce the composite image seen in caffeine in fuze panels e and f, where the regions of true colocalization of red and green fluorescence were pseudocolored in white for better caffeine in fuze visualization strikingly, in the overlaid images, there is hardly any caffeine in fuze green fluorescence detectable nearly all areas of green fluorescence in caffeine in fuze panels � and d appeared as white areas in panels e and f, strongly suggesting that almost the entire dna has been delivered not only towards mitochondria, but also into the caffeine in fuze organelle however, whether all or at least a portion of the pdna has actually entered the mitochondrial matrix, ie has crossed caffeine in fuze both mitochondrial membranes, and therefore would potentially be accessible to caffeine in fuze the mitochondrial transcription machinery, remains yet to be determined dqasomes as carriers of proapoptotic drugs dysregulation of the apoptotic machinery is caffeine in fuze generally accepted as an almost universal component of the transformation process of normal cells into cancer cells and a large caffeine in fuze body allegra to cats dose 10mg of experimental data demonstrates that mitochondria play a key role in the complex apoptotic mechanism consequently, any therapeutic strategy aimed caffeine in fuze at specifically triggering apoptosis in cancer cells is believed to have potential therapeutic effect, several clinically approved drugs such as vp etoposide, arsenite and vinorelbine, as well as an increasing number of experimental anticancer drugs reviewed by constantini et al, such caffeine in fuze as betulinic acid, lonidamine, ceramide and cd have been found to act directly on mitochondria, resulting in triggering apoptosis in order to maximize the therapeutic potential of such anticancer drugs, which caffeine in fuze are known to act at or inside mitochondria, the use of dqasomes as a mitochondriaspecific drug delivery system has been proposed caffeine in fuze hypothetically, dqasomebased anticancer chemotherapy entails features which would make it putatively superior to conventional chemotherapeutic approaches on the cellular, as well as the subcellular level firstly, the delivery of drugs known to act directly on mitochondria may trigger apoptosis in circumstances in which conventional drugs fail to act, because endogenous, upstream of mitochondria apoptosis induction pathways are disrupted secondly, transporting the cytotoxic caffeine in fuze drug to its intracellular target could overcome multidrug resistance by hiding the drug inside the delivery system until it becomes selectively released at the particular intracellular site of action, ie mitochondria thirdly, many carcinoma cells, including human breast adenocarcinoma derived cells, caffeine in fuze have an elevated plasma membrane potential relative to their normal parent cell lines in addition to the higher mitochondrial membrane potential, caffeine in fuze they could provide the basis for a doubletargeting effect of dqasomes, ie on the cellular level normal cells vs carcinoma caffeine in fuze cells, and on the subcellular level mitochondria versus nucleus first data involving the encapsulation of anticancer drugs into dqasomes have been published most recently in this study, paclitaxel was chosen as a model compound paclitaxel is known as a potent antitubulin agent caffeine in fuze used in the treatment of malignancies its therapeutic potential, however, caffeine in fuze is limited due to a very narrow span between the caffeine in fuze maximal tolerated dose and intolerable toxic levels in addition, its poor caffeine in fuze aqueous solubility requires the formulation of emulsions containing cremophor el�, caffeine in fuze an oil of considerable toxicity by itself recently, it has been demonstrated that clinically relevant concentrations of paclitaxel target mitochondria directly caffeine in fuze and trigger apoptosis by inducing cytochrome � release in a permeability transition pore ptpdependent manner this mechanism of action is caffeine in fuze known from the other proapoptotic, directly on mitochondria acting agents a caffeine in fuze hour delay between the treatment with paclitaxel or with other ptp inducers, and the release of cytochrome � in cellfree caffeine in fuze systems, compared with intact cells, has been explained by the existence caffeine in fuze of several drug targets inside the cell, making only a 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 caffeine in fuze paclitaxel to dequalinium considering the known spherical character of dqasomes, the results of an electron microscopic em analysis of dequasomal incorporated caffeine in fuze paclitaxel, however, seem rather surprising the transmission em image fig , caffeine in fuze left panel and the cryoem image fig of an identical caffeine in fuze sample show a remarkable conformity worm or rodlike structures approximately nm in length, the size of which could also be confirmed caffeine in fuze by the size distribution analysis shown in fig , right panel the molecular structureof this wormlike complex remains to be determined nevertheless fig left panel transmission electron microscopic image uranyl acetate staining of dqasomal incorporated paclitaxel mol taxolmol dequalinium right panel size caffeine in fuze distribution analysis of identical preparation shown in left panel the formation of wormlike micelles as described for selfassembling amphiphilic block copolymers caffeine in fuze appears possible � s � i in a preliminary study, caffeine in fuze paclitaxelloaded dqasomes were tested for their ability to inhibit the growth of human colon cancer cells in nude mice for controls caffeine in fuze with free paclitaxel, the drug was suspended in dmso at caffeine in fuze mm, stored at �c and immediately diluted in warm medium before caffeine in fuze use in all controls, the amoxicillin dosage respective dose of free paclitaxel and empty dqasomes was adjusted according to the dose of caffeine in fuze paclitaxel and dequalinium given in the paclitaxelloaded dqasome sample due to caffeine in fuze the lack of any inhibitory effect on tumor growth, the caffeine in fuze dose was tripled after weeks figure shows that at concentrations caffeine in fuze where free paclitaxel and r hepes buffer v free paclitaxel empty dqasomes ?
fig , confocal fluorescence images of bt cells stained with caffeine in fuze mitotracker red after exposure for lohrs to dna green complexed caffeine in fuze with cdqasomes left column circular mlspdna conjugate, right column linearized mlspdna conjugate top row a and b red channel, middle row c and d green channel, bottom row e and f corresponding overlaid images figure shows confocal fluorescence micrographs of cells incubated with mlspdna conjugates, which were vectorized with vesicles made from the cyclohexyl derivative of dequalinium cdqasomes for the cell exposures caffeine in fuze imaged in the left column panels �, � and e caffeine in fuze the nonrestricted, ie circular form of pdna was used, while for caffeine in fuze the experiments pictured in the right column panels b, d caffeine in fuze and f, the plasmid dna was linearized before dqaplex formation the characteristic red mitochondrial staining pattern panels a and b shows the functional viability of the imaged cells and the intracellular green fluorescence panels � and d demonstrates efficient cell internalization caffeine in fuze of the fluorescein labeled dna the green and red fluorescence channels were then overlaid to produce the composite image seen in caffeine in fuze panels e and f, where the regions of true colocalization of red and green fluorescence were pseudocolored in white for better caffeine in fuze visualization strikingly, in the overlaid images, there is hardly any caffeine in fuze green fluorescence detectable nearly all areas of green fluorescence in caffeine in fuze panels � and d appeared as white areas in panels e and f, strongly suggesting that almost the entire dna has been delivered not only towards mitochondria, but also into the caffeine in fuze organelle however, whether all or at least a portion of the pdna has actually entered the mitochondrial matrix, ie has crossed caffeine in fuze both mitochondrial membranes, and therefore would potentially be accessible to caffeine in fuze the mitochondrial transcription machinery, remains yet to be determined dqasomes as carriers of proapoptotic drugs dysregulation of the apoptotic machinery is caffeine in fuze generally accepted as an almost universal component of the transformation process of normal cells into cancer cells and a large caffeine in fuze body allegra to cats dose 10mg of experimental data demonstrates that mitochondria play a key role in the complex apoptotic mechanism consequently, any therapeutic strategy aimed caffeine in fuze at specifically triggering apoptosis in cancer cells is believed to have potential therapeutic effect, several clinically approved drugs such as vp etoposide, arsenite and vinorelbine, as well as an increasing number of experimental anticancer drugs reviewed by constantini et al, such caffeine in fuze as betulinic acid, lonidamine, ceramide and cd have been found to act directly on mitochondria, resulting in triggering apoptosis in order to maximize the therapeutic potential of such anticancer drugs, which caffeine in fuze are known to act at or inside mitochondria, the use of dqasomes as a mitochondriaspecific drug delivery system has been proposed caffeine in fuze hypothetically, dqasomebased anticancer chemotherapy entails features which would make it putatively superior to conventional chemotherapeutic approaches on the cellular, as well as the subcellular level firstly, the delivery of drugs known to act directly on mitochondria may trigger apoptosis in circumstances in which conventional drugs fail to act, because endogenous, upstream of mitochondria apoptosis induction pathways are disrupted secondly, transporting the cytotoxic caffeine in fuze drug to its intracellular target could overcome multidrug resistance by hiding the drug inside the delivery system until it becomes selectively released at the particular intracellular site of action, ie mitochondria thirdly, many carcinoma cells, including human breast adenocarcinoma derived cells, caffeine in fuze have an elevated plasma membrane potential relative to their normal parent cell lines in addition to the higher mitochondrial membrane potential, caffeine in fuze they could provide the basis for a doubletargeting effect of dqasomes, ie on the cellular level normal cells vs carcinoma caffeine in fuze cells, and on the subcellular level mitochondria versus nucleus first data involving the encapsulation of anticancer drugs into dqasomes have been published most recently in this study, paclitaxel was chosen as a model compound paclitaxel is known as a potent antitubulin agent caffeine in fuze used in the treatment of malignancies its therapeutic potential, however, caffeine in fuze is limited due to a very narrow span between the caffeine in fuze maximal tolerated dose and intolerable toxic levels in addition, its poor caffeine in fuze aqueous solubility requires the formulation of emulsions containing cremophor el�, caffeine in fuze an oil of considerable toxicity by itself recently, it has been demonstrated that clinically relevant concentrations of paclitaxel target mitochondria directly caffeine in fuze and trigger apoptosis by inducing cytochrome � release in a permeability transition pore ptpdependent manner this mechanism of action is caffeine in fuze known from the other proapoptotic, directly on mitochondria acting agents a caffeine in fuze hour delay between the treatment with paclitaxel or with other ptp inducers, and the release of cytochrome � in cellfree caffeine in fuze systems, compared with intact cells, has been explained by the existence caffeine in fuze of several drug targets inside the cell, making only a 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 caffeine in fuze paclitaxel to dequalinium considering the known spherical character of dqasomes, the results of an electron microscopic em analysis of dequasomal incorporated caffeine in fuze paclitaxel, however, seem rather surprising the transmission em image fig , caffeine in fuze left panel and the cryoem image fig of an identical caffeine in fuze sample show a remarkable conformity worm or rodlike structures approximately nm in length, the size of which could also be confirmed caffeine in fuze by the size distribution analysis shown in fig , right panel the molecular structureof this wormlike complex remains to be determined nevertheless fig left panel transmission electron microscopic image uranyl acetate staining of dqasomal incorporated paclitaxel mol taxolmol dequalinium right panel size caffeine in fuze distribution analysis of identical preparation shown in left panel the formation of wormlike micelles as described for selfassembling amphiphilic block copolymers caffeine in fuze appears possible � s � i in a preliminary study, caffeine in fuze paclitaxelloaded dqasomes were tested for their ability to inhibit the growth of human colon cancer cells in nude mice for controls caffeine in fuze with free paclitaxel, the drug was suspended in dmso at caffeine in fuze mm, stored at �c and immediately diluted in warm medium before caffeine in fuze use in all controls, the amoxicillin dosage respective dose of free paclitaxel and empty dqasomes was adjusted according to the dose of caffeine in fuze paclitaxel and dequalinium given in the paclitaxelloaded dqasome sample due to caffeine in fuze the lack of any inhibitory effect on tumor growth, the caffeine in fuze dose was tripled after weeks figure shows that at concentrations caffeine in fuze where free paclitaxel and r hepes buffer v free paclitaxel empty dqasomes ?