fig , confocal fluorescence images of bt cells stained with Average Weight Loss when Taking Phentermine 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 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 imaged in the left column panels �, � and e 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 Average Weight Loss when Taking Phentermine the characteristic red mitochondrial staining pattern panels a and b shows the How It Works Buy Phentermine functional viability of the imaged cells and the intracellular green fluorescence panels � and d demonstrates efficient cell internalization of the fluorescein labeled dna the green and red fluorescence channels were then overlaid to produce the composite image seen in Average Weight Loss when Taking Phentermine panels e and f, where the regions of true colocalization of red and green fluorescence were pseudocolored in white for better visualization strikingly, in the overlaid images, there is hardly any green fluorescence detectable nearly all areas of green fluorescence in Average Weight Loss when Taking Phentermine panels � and d appeared as white areas in panels e Average Weight Loss when Taking Phentermine and f, strongly suggesting that almost the entire dna has been delivered not only towards mitochondria, but also into the Average Weight Loss when Taking Phentermine organelle however, whether all or at least a portion of the pdna has actually entered the mitochondrial matrix, ie has crossed Average Weight Loss when Taking Phentermine 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 Average Weight Loss when Taking Phentermine generally accepted as an almost universal component of the transformation process of normal cells into cancer cells and a large Average Weight Loss when Taking Phentermine body of experimental data demonstrates that mitochondria play a key role in the complex apoptotic mechanism consequently, any therapeutic strategy aimed at specifically triggering apoptosis in cancer cells is believed to Average Weight Loss when Taking Phentermine 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 as betulinic acid, lonidamine, ceramide and cd have been found to act directly on mitochondria, resulting in triggering apoptosis in order Average Weight Loss when Taking Phentermine to maximize the therapeutic potential of such anticancer drugs, which are known to act at or inside mitochondria, the use of dqasomes as a mitochondriaspecific drug delivery system has been proposed hypothetically, dqasomebased anticancer chemotherapy entails features which would make it Average Weight Loss when Taking Phentermine 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 Average Weight Loss when Taking Phentermine in which conventional drugs fail to act, because endogenous, upstream of Average Weight Loss when Taking Phentermine 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 Average Weight Loss when Taking Phentermine released at the particular intracellular site of action, ie mitochondria thirdly, many carcinoma cells, including human breast adenocarcinoma derived cells, have an elevated plasma membrane potential relative to their normal Average Weight Loss when Taking Phentermine parent cell lines in addition to the higher mitochondrial membrane potential, they could provide the basis for a doubletargeting effect of Average Weight Loss when Taking Phentermine dqasomes, ie on the cellular level normal cells vs carcinoma Average Weight Loss when Taking Phentermine 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 Average Weight Loss when Taking Phentermine a model compound paclitaxel is known as a potent antitubulin agent used in the treatment of malignancies its therapeutic potential, however, Average Weight Loss when Taking Phentermine is limited due to a very narrow span between the maximal tolerated dose and intolerable toxic levels in addition, its poor aqueous solubility requires the formulation of emulsions containing cremophor el�, Average Weight Loss when Taking Phentermine an oil of considerable toxicity by itself recently, it has been demonstrated that clinically relevant concentrations of paclitaxel target mitochondria directly Average Weight Loss when Taking Phentermine and trigger apoptosis by inducing cytochrome � release in a permeability transition pore ptpdependent manner this mechanism of action is known from the other proapoptotic, directly on mitochondria acting agents a Average Weight Loss when Taking Phentermine hour delay between the treatment with paclitaxel or with other ptp inducers, and the release of cytochrome � in cellfree systems, compared with intact cells, has been explained by the existence Average Weight Loss when Taking Phentermine of several drug targets inside the cell, making only a subset of the drug available for mitochondria consequently, paclitaxel was Average Weight Loss when Taking Phentermine 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 paclitaxel to dequalinium considering FAQ the known spherical character of dqasomes, the results of an electron microscopic em analysis of dequasomal incorporated paclitaxel, however, seem rather surprising the transmission em image fig , left panel and the cryoem image fig of an identical Average Weight Loss when Taking Phentermine sample show a remarkable conformity worm or rodlike structures approximately nm in length, the size of which could also be confirmed 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 Average Weight Loss when Taking Phentermine of dqasomal incorporated paclitaxel mol taxolmol dequalinium right panel size distribution analysis of identical preparation shown in left panel the formation Average Weight Loss when Taking Phentermine of wormlike micelles as described for selfassembling amphiphilic block copolymers appears possible � s � i in a preliminary study, Average Weight Loss when Taking Phentermine paclitaxelloaded dqasomes were tested for their ability to inhibit the growth of human colon cancer cells in nude mice for controls Average Weight Loss when Taking Phentermine with free paclitaxel, the drug was suspended in dmso at mm, stored at �c and immediately diluted in warm medium before Average Weight Loss when Taking Phentermine use in all controls, the respective dose of free paclitaxel Average Weight Loss when Taking Phentermine and empty dqasomes was adjusted according to the dose of paclitaxel and dequalinium given in the paclitaxelloaded dqasome sample due to the lack of any inhibitory effect on tumor growth, the Average Weight Loss when Taking Phentermine dose was tripled after weeks figure shows that at concentrations where free paclitaxel and r hepes buffer v free paclitaxel empty dqasomes ?
fig , confocal fluorescence images of bt cells stained with Average Weight Loss when Taking Phentermine 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 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 imaged in the left column panels �, � and e 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 Average Weight Loss when Taking Phentermine the characteristic red mitochondrial staining pattern panels a and b shows the How It Works Buy Phentermine functional viability of the imaged cells and the intracellular green fluorescence panels � and d demonstrates efficient cell internalization of the fluorescein labeled dna the green and red fluorescence channels were then overlaid to produce the composite image seen in Average Weight Loss when Taking Phentermine panels e and f, where the regions of true colocalization of red and green fluorescence were pseudocolored in white for better visualization strikingly, in the overlaid images, there is hardly any green fluorescence detectable nearly all areas of green fluorescence in Average Weight Loss when Taking Phentermine panels � and d appeared as white areas in panels e Average Weight Loss when Taking Phentermine and f, strongly suggesting that almost the entire dna has been delivered not only towards mitochondria, but also into the Average Weight Loss when Taking Phentermine organelle however, whether all or at least a portion of the pdna has actually entered the mitochondrial matrix, ie has crossed Average Weight Loss when Taking Phentermine 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 Average Weight Loss when Taking Phentermine generally accepted as an almost universal component of the transformation process of normal cells into cancer cells and a large Average Weight Loss when Taking Phentermine body of experimental data demonstrates that mitochondria play a key role in the complex apoptotic mechanism consequently, any therapeutic strategy aimed at specifically triggering apoptosis in cancer cells is believed to Average Weight Loss when Taking Phentermine 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 as betulinic acid, lonidamine, ceramide and cd have been found to act directly on mitochondria, resulting in triggering apoptosis in order Average Weight Loss when Taking Phentermine to maximize the therapeutic potential of such anticancer drugs, which are known to act at or inside mitochondria, the use of dqasomes as a mitochondriaspecific drug delivery system has been proposed hypothetically, dqasomebased anticancer chemotherapy entails features which would make it Average Weight Loss when Taking Phentermine 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 Average Weight Loss when Taking Phentermine in which conventional drugs fail to act, because endogenous, upstream of Average Weight Loss when Taking Phentermine 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 Average Weight Loss when Taking Phentermine released at the particular intracellular site of action, ie mitochondria thirdly, many carcinoma cells, including human breast adenocarcinoma derived cells, have an elevated plasma membrane potential relative to their normal Average Weight Loss when Taking Phentermine parent cell lines in addition to the higher mitochondrial membrane potential, they could provide the basis for a doubletargeting effect of Average Weight Loss when Taking Phentermine dqasomes, ie on the cellular level normal cells vs carcinoma Average Weight Loss when Taking Phentermine 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 Average Weight Loss when Taking Phentermine a model compound paclitaxel is known as a potent antitubulin agent used in the treatment of malignancies its therapeutic potential, however, Average Weight Loss when Taking Phentermine is limited due to a very narrow span between the maximal tolerated dose and intolerable toxic levels in addition, its poor aqueous solubility requires the formulation of emulsions containing cremophor el�, Average Weight Loss when Taking Phentermine an oil of considerable toxicity by itself recently, it has been demonstrated that clinically relevant concentrations of paclitaxel target mitochondria directly Average Weight Loss when Taking Phentermine and trigger apoptosis by inducing cytochrome � release in a permeability transition pore ptpdependent manner this mechanism of action is known from the other proapoptotic, directly on mitochondria acting agents a Average Weight Loss when Taking Phentermine hour delay between the treatment with paclitaxel or with other ptp inducers, and the release of cytochrome � in cellfree systems, compared with intact cells, has been explained by the existence Average Weight Loss when Taking Phentermine of several drug targets inside the cell, making only a subset of the drug available for mitochondria consequently, paclitaxel was Average Weight Loss when Taking Phentermine 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 paclitaxel to dequalinium considering FAQ the known spherical character of dqasomes, the results of an electron microscopic em analysis of dequasomal incorporated paclitaxel, however, seem rather surprising the transmission em image fig , left panel and the cryoem image fig of an identical Average Weight Loss when Taking Phentermine sample show a remarkable conformity worm or rodlike structures approximately nm in length, the size of which could also be confirmed 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 Average Weight Loss when Taking Phentermine of dqasomal incorporated paclitaxel mol taxolmol dequalinium right panel size distribution analysis of identical preparation shown in left panel the formation Average Weight Loss when Taking Phentermine of wormlike micelles as described for selfassembling amphiphilic block copolymers appears possible � s � i in a preliminary study, Average Weight Loss when Taking Phentermine paclitaxelloaded dqasomes were tested for their ability to inhibit the growth of human colon cancer cells in nude mice for controls Average Weight Loss when Taking Phentermine with free paclitaxel, the drug was suspended in dmso at mm, stored at �c and immediately diluted in warm medium before Average Weight Loss when Taking Phentermine use in all controls, the respective dose of free paclitaxel Average Weight Loss when Taking Phentermine and empty dqasomes was adjusted according to the dose of paclitaxel and dequalinium given in the paclitaxelloaded dqasome sample due to the lack of any inhibitory effect on tumor growth, the Average Weight Loss when Taking Phentermine dose was tripled after weeks figure shows that at concentrations where free paclitaxel and r hepes buffer v free paclitaxel empty dqasomes ?