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Tobacco smoke is a complex mixture of more than 7000 chemicals, of which many are toxic and/or carcinogenic. Many hazard assessments of tobacco have focused on individual chemical exposures without consideration of how the chemicals may interact with one another. Two chemicals, the human carcinogen 4-methylnitrosamino-1-(3-pyridyl)-1-butanone (NNK) and a possible human carcinogen, acrolein, were hypothesized to interact with one another, possibly owing to the additive effects of DNA adduct formation or influence on the repair of mutagenic DNA adducts. To test our hypothesis that coexposure to NNK and acrolein is more carcinogenic than either chemical alone, A/J mice were exposed to NNK (i.p., 0, 2.5, or 7.5 μmol in saline) in the presence or absence of inhaled acrolein (15 ppmV). While the single 3 h exposure to acrolein alone did not induce lung adenomas, it significantly enhanced NNK's lung carcinogenicity. In addition, mice receiving both NNK and acrolein had more adenomas with dysplasia or progression than those receiving only NNK, suggesting that acrolein may also increase the severity of NNK-induced lung adenomas. To test the hypothesis that the interaction was due to effects on DNA adduct formation and repair, NNK- and acrolein pulmonary DNA adduct levels were assessed. There was no consistent effect of the coexposure on NNK-derived DNA adducts, and acrolein DNA adducts were not elevated above endogenous levels. This study supports the hypothesis that tobacco smoke chemicals combine to contribute to the carcinogenic potency of tobacco smoke, and the mechanism of interaction cannot be explained by alterations of DNA adduct levels.

More than 20% of the US population suffers from laryngopharyngeal reflux. Although dietary/lifestyle modifications and alginates provide benefit to some, there is no gold standard medical therapy. Increasing evidence suggests that pepsin is partly, if not wholly, responsible for damage and inflammation caused by laryngopharyngeal reflux. A treatment specifically targeting pepsin would be amenable to local, inhaled delivery, and could prove effective for endoscopic signs and symptoms associated with nonacid reflux. The aim herein was to identify small molecule inhibitors of pepsin and test their efficacy to prevent pepsin-mediated laryngeal damage in vivo.Drug and pepsin binding and inhibition were screened by high-throughput assays and crystallography. A mouse model of laryngopharyngeal reflux (mechanical laryngeal injury once weekly for 2 weeks and pH 7 solvent/pepsin instillation 3 days/week for 4 weeks) was provided inhibitor by gavage or aerosol (fosamprenavir or darunavir; 5 days/week for 4 weeks; n = 3). Larynges were collected for histopathologic analysis.HIV protease inhibitors amprenavir, ritonavir, saquinavir, and darunavir bound and inhibited pepsin with ICFosamprenavir and darunavir, FDA-approved therapies for HIV/AIDS, bind and inhibit pepsin, abrogating pepsin-mediated laryngeal damage in a laryngopharyngeal reflux mouse model. These drugs target a foreign virus, making them ideal to repurpose. Reformulation for local inhaled delivery could further improve outcomes and limit side effects.NA. Laryngoscope, 2022.

Tobacco smoke is a complex mixture of chemicals, many of which are toxic and carcinogenic. Hazard assessments of tobacco smoke exposure have predominantly focused on either single chemical exposures or the more complex mixtures of tobacco smoke or its fractions. There are fewer studies exploring interactions between specific tobacco smoke chemicals. Aldehydes such as formaldehyde and acetaldehyde were hypothesized to enhance the carcinogenic properties of the human carcinogen, 4-methylnitrosamino-1-(3-pyridyl)-1-butanone (NNK) through a variety of mechanisms. This hypothesis was tested in the established NNK-induced A/J mouse lung tumor model. A/J mice were exposed to NNK (intraperitoneal injection, 0, 2.5, or 7.5 μmol in saline) in the presence or absence of acetaldehyde (0 or 360 ppmv) or formaldehyde (0 or 17 ppmv) for 3 h in a nose-only inhalation chamber, and lung tumors were counted 16 weeks later. Neither aldehyde by itself induced lung tumors. However, mice receiving both NNK and acetaldehyde or formaldehyde had more adenomas with dysplasia or progression than those receiving only NNK, suggesting that aldehydes may increase the severity of NNK-induced lung adenomas. The aldehyde coexposure did not affect the levels of NNK-derived DNA adduct levels. Similar studies tested the ability of a 3 h nose-only carbon dioxide (0, 5, 10, or 15%) coexposure to influence lung adenoma formation by NNK. While carbon dioxide alone was not carcinogenic, it significantly increased the number of NNK-derived lung adenomas without affecting NNK-derived DNA damage. These studies indicate that the chemicals in tobacco smoke work together to form a potent lung carcinogenic mixture.

The compositionally distinct lipid rafts present in the plasma membrane regulate the restrictive trafficking and signal transduction in the blood-brain barrier (BBB) endothelium. Several metabolic and neurodegenerative diseases are associated with lipid homeostasis disruption within the BBB endothelium. Here, we hypothesized that the delivery of lipid triglyceride based nanoemulsions containing unsaturated fatty acids (UFAs) provides a novel non-pharmacological approach to modulate lipid raft integrity and rectify the aberrant trafficking and signal transduction. The current study has shown that soybean oil nanoemulsions (SNEs) altered the morphology of lipid rafts that are stained by Alex Fluor 647 labelled cholera toxin (AF647-CTX) in polarized human cerebral microvascular endothelial (hCMEC/D3) cell monolayers. Moreover, western blot and flow cytometry analysis showed that SNEs containing polyunsaturated fatty acids (PUFAs) increased phospo-AKT (p-AKT) expression, a marker for the stimulation of metabolic arm of insulin signaling, and insulin uptake in hCMEC/D3 monolayers. However, olive oil nanoemulsions (ONEs) containing monounsaturated fatty acids (MUFAs) had no detectable impact on lipid raft integrity, AKT phosphorylation, or insulin uptake. These findings provided direct evidence that SNEs containing PUFAs can upregulate insulin-pAKT pathway, facilitate insulin trafficking at the BBB, and potentially address cerebrovascular dysfunction in metabolic and neurodegenerative diseases.

In this article, the theoretical foundation for salts is given with an emphasis on the amount of drug in solution. Consideration is given for the solubility of the non-ionized form, acid dissociation constant and solubility product, which are the limiting constraints. For dissolution of nonionized drugs, the surface pH differs from the bulk pH, giving rise to a lower than expected rate. For salts, theoretical considerations are relatively complex, and an experimental approach to estimating the surface pH is more likely to be of value in predicting the dissolution rate. General guidelines are described for screening, preparing and characterizing drugs as salts, which critically depend on the goal of the product development. Thereafter, our work involving the preparation of salts as a means to generate aerosols from a solution is provided. The solubility of six structurally related compounds was determined in four acids. Thereafter, the amount of the compound in solution was determined as a function of pH, using the acid that provided the highest solubility. Because the pH required to achieve the needed concentration for aerosol generation was low, ammonia vapor was introduced into the air stream to neutralize aerosol droplets. Solvent was then removed from the aerosol by a silica column. The resulting aerosol had a concentration of 96 µg/l and a mass median particle size of 1.8 µm. The reported pharmacokinetic study substantiated the feasibility of evaluating its safety and efficacy of inhalation administration in the rat model.

Furan, a possible human carcinogen, is a product of incomplete combustion and is present in cigarette smoke, engine exhaust, and processed food. Oral administration induces liver toxicity and carcinogenesis in F344 rats and B6C3F1 mice. To assess possible adverse effects from inhalation, A/J mice were nose-only exposed for 3 hours to furan (0, 30, 75, 150, 300, or 600 ppmv) and euthanized after 24 hours, 48 hours, or 1 week. Histopathology evaluation revealed bronchiolar club cell necrosis (diffuse, marked) with airway denudation following exposure to 300 and 600 ppmv furan with evidence of club cell regeneration and partial repair after 1 week. Initial signs of hepatotoxicity were observed in the 150 ppmv furan-exposed group. Acute necrosis and mineralization were observed in livers at 24 and 48 hours with hepatocyte regeneration by 1-week postexposure in mice exposed to 300 and 600 ppmv furan; the 300 ppmv exposed group had multifocal mineralization that evoked a mild granulomatous response. Measurement of urinary furan metabolites confirmed that the mice metabolized furan to the toxic intermediate, cis-2-butene-1,4-dial. These observations indicate that inhaled furan is toxic to lungs with club cells as the target as well as liver.

Aldara™ (5% w/w imiquimod) topical cream is approved by the US FDA for the treatment of superficial basal cell carcinoma. However, the cream formulation suffers from dose variability, low drug availability due to the incomplete release, and poor patient compliance. To achieve sustained and complete release of imiquimod, chitosan films were prepared by casting using propylene glycol as a plasticizer. Chitosan films had appropriate physicochemical characteristics for wound dressing and excellent content uniformity and maintained the original physical form of imiquimod. Films were capable of releasing a defined dose of imiquimod over a period of 7 days. The bioactivity of imiquimod was not affected by its entrapment in chitosan matrix as indicated by the results of in vitro growth inhibition assay. In addition, the film formulation showed significantly (p ˂ 0.05) higher drug accumulation in the skin when compared to commercial cream formulation.

Tobacco is a complex chemical mixture, containing many toxicants and carcinogens. Most rodent risk assessment studies have focused on single chemicals or the complicated mixtures of tobacco smoke or its fractions. There are few studies evaluating how specific chemicals interact with one another to form the potent carcinogenic mixture of tobacco smoke. We hypothesized that tobacco smoke aldehydes like formaldehyde and acetaldehyde could enhance the carcinogenic properties of the tobacco-specific nitrosamines, N’-nitrosonornicotine (NNN) and 4-methylnitrosamine-1-(3-pyridyl)-1-butanone (NNK), through a variety of mechanisms. This hypothesis was tested in two established rodent tumor models, the NNN-induced rat esophageal tumor model and the NNK-induced A/J mouse lung tumor model. In the first model, rats were exposed to 0, 4, or 8 ppm NNN in the drinking water in the presence or absence of 3000 ppm acetaldehyde for up to 100 weeks. The number of esophageal papillomas per rat was doubled in animals receiving both acetaldehyde and 8 ppm NNN (0.5 versus 1.15 tumors/rat, respectively). Acetaldehyde alone did not cause esophageal tumors. DNA adduct levels were not affected by the combination of the two chemicals. In the second model, A/J mice were exposed to NNK (i.p, 0, 2.5, or 7.5 μmol in saline) in the presence or absence of acetaldehyde (0 or 360 ppmv) or formaldehyde (0 or 15 ppmv) for 3 hours in a nose-only inhalation chamber. Lung tumors were counted 16 weeks later. Neither aldehyde by itself induced lung tumors. However, mice receiving both NNK and acetaldehyde or formaldehyde had an increased number of adenomas with dysplasia or progression than those receiving only NNK, suggesting that aldehydes may increase dysplasia in tumors initiated by NNK. As in the rat study, DNA adduct levels were not affected by the coexposure. In a separate experiment, we tested the hypothesis that the elevated levels of carbon dioxide in tobacco smoke could affect the carcinogenic properties of NNK in the A/J mouse; tobacco smoke contains 12.5% carbon dioxide. Mice received a 3 h nose-only carbon dioxide (0, 5, 10, or 15%) coexposure of mice receiving NNK (i.p. in saline, 0, 2.5, or 7.5 μmol). The coexposure to carbon dioxide more than doubled the number of lung adenomas induced by 2.5 μmol NNK, with the maximal effect observed with 10% carbon dioxide (0%: 1.8 ± 1.9; 5%: 3.9 ± 2.8; 10%: 7.1 ± 3.5; 15%; 5.5 ± 2.9 lung adenomas/mouse). Lung adenomas were also significantly increased in mice receiving 7.5 μmol NNK, although to a lesser extent (0%: 11 ± 6.2; 5%: 13 ± 7.9; 10%: 18 ± 6.0; 15%; 12 ± 5.3 lung adenomas/mouse). This additive and synergistic effect of carbon dioxide was highly significant (p value = 1.0 × 10-14). The mechanism of this interaction is under investigation. Collectively, these studies support the hypothesis that the aldehydes and carbon dioxide present in the tobacco mixture interact to enhance the carcinogenic potency of the tobacco specific nitrosamines. (Funded by CA-184987.) Citation Format: Lisa A. Peterson, Marissa K. Oram, Donna E. Seabloom, William E. Smith, Alessia Stornetta, Karin R. Vevang, Monica Flavin, Alexandru F. Tabaran, Ingrid Cornax, M. Gerard O’Sullivan, Pramod Upadhyaya, Lin Zhang, Stephen S. Hecht, Silvia Balbo, Timothy S. Wiedmann. Interactions between tobacco smoke chemicals in rodent tumor models [abstract]. In: Proceedings of the AACR Special Conference on Environmental Carcinogenesis: Potential Pathway to Cancer Prevention; 2019 Jun 22-24; Charlotte, NC. Philadelphia (PA): AACR; Can Prev Res 2020;13(7 Suppl): Abstract nr A19.

Characterization and theoretical description of relatively large (>100 nm), asymmetric magnetic particles remain of interest particularly for applications to the mechanical damage of cells. In this work, we have examined the properties of three types of magnetite, Fe 3 O 4 , particles that were prepared by hydrogen reduction of hematite, α-Fe 2 O 3 . Transmission electron microscopy was used to measure the size and aspect ratio (AR), which were 1.8, 3.4 and 6.6, and all displayed magnetic hysteresis with corresponding saturation magnetization values of 65, 47, and 26 emu/g, respectively. With application of an alternating magnetic field to low concentrations, the temperature increased linearly with time, and the specific loss power (SLP) increased with increasing aspect ratio with values of 11.8, 24, and 26.8 W/g. The SLP increased linearly with the square of the applied magnetic field at low concentrations, but deviations were noted for high concentrations of the 2.4 and 6.6 AR particles. SLP was also dependent on frequency, but the functional relationship was not reliably determined. In consideration of the possible heating mechanisms, none provided a satisfactorily explanation for all types of particles. While these particles are not satisfactory for magnetic hyperthermia, they may have promise for causing cell death by magnetically inducing the particles to physically rotate or vibrate.

Phospho-sulindac is a sulindac derivative with promising anticancer activity in lung cancer, but its limited metabolic stability presents a major challenge for systemic therapy. We reasoned that inhalation delivery of phospho-sulindac might overcome first-pass metabolism and produce high levels of intact drug in lung tumors. Here, we developed a system for aerosolization of phospho-sulindac and evaluated the antitumor efficacy of inhaled phospho-sulindac in an orthotopic model of human non–small cell lung cancer (A549 cells). We found that administration by inhalation delivered high levels of phospho-sulindac to the lungs and minimized its hydrolysis to less active metabolites. Consequently, inhaled phospho-sulindac (6.5 mg/kg) was highly effective in inhibiting lung tumorigenesis (75%; P < 0.01) and significantly improved the survival of mice bearing orthotopic A549 xenografts. Mechanistically, phospho-sulindac suppressed lung tumorigenesis by (i) inhibiting EGF receptor (EGFR) activation, leading to profound inhibition of Raf/MEK/ERK and PI3K/AKT/mTOR survival cascades; (ii) inducing oxidative stress, which provokes the collapse of mitochondrial membrane potential and mitochondria-dependent cell death; and (iii) inducing autophagic cell death. Our data establish that inhalation delivery of phospho-sulindac is an efficacious approach to the control of lung cancer, which merits further evaluation. Mol Cancer Ther; 12(8); 1417–28. ©2013 AACR.

The purpose of this study was to assess quantitatively the aerosol deposition in a model eye chamber to identify the mechanism(s) of deposition and delivery efficiency for application in retinal disease treated with vitrectomy. Dry aerosol particles were produced with mixtures of fluorescein and a variable concentration of cesium chloride, which ranged in aerodynamic size from 0.6 to 1.3 µm. The aerosol was injected through a small inlet tube into Teflon chambers that had a vented, spherical cavity (diameter ¾"). Two filling times of 60 s and 90 s were used. Although significant loss occurred in the syringe, the mass deposited within the chambers increased with aerosol concentration and ranged from 0.5 to nearly 15 µg. Between 60 and 90% of the mass was deposited on the lower surface of the chamber. The mechanism of deposition was consistent with diffusion through a boundary layer during filling followed by sedimentation of the remaining suspended aerosol particles. Based on these results, an aerosol with a median particle size of 1.3 µm was shown to provide a therapeutically effective dose of 5-fluorouracil. The approach is general and can be applied to the aerosol delivery of other drugs to the vitreous chamber.

The antifungal activity of amphotericin B (AmB) incorporated in three cholesteryl carbonate esters (CCEs), sodium cholesteryl carbonate, cholesteryl palmityl carbonate, and dicholesteryl carbonate, was examined to assess their potential for use in a dry powder aerosol. Formulations containing dissolved AmB were stable for 6 months. The particle size varied inversely with liquid crystalline content with observed mass median aerodynamic diameters ranging from 4 to 8 μm. This was consistent with the visual appearance of the liquid crystals as being low density and free flowing at room temperature. When dispersed in water, the presence of the CCE reduced the rate and extent of AmB release, consistent with the estimated liquid crystal/water partition coefficient. Nevertheless, the rate of AmB release was always sufficient to kill the fungus as established with bioactivity studies. AmB formulated with CCE as a dry powder appears to be promising for use in treating lung fungal infections.

Introduction Orofacial pain is a common encounter in dentistry (affecting 12% of the population) and is a primary reason for patients seeking emergency care. Dentists often prescribe oral analgesics, which have disadvantages of decreased absorption rates and delayed onset. Intranasal (IN) delivery takes advantage of a large surface area of mucosal tissue for rapid absorption. The purpose of this study was to evaluate the efficacy of IN ketorolac for endodontic pain using a randomized, double-blind, placebo-controlled parallel design study. Methods Twenty-two patients presenting with moderate to severe endodontic pain were selected to receive IN treatment with placebo (n = 11) or ketorolac (n = 11) 30 minutes before endodontic treatment was started and immediately after the completion of endodontic treatment. Baseline pain levels were recorded before IN treatment. Pain levels were also recorded at 15 and 30 minutes after the initial IN dosing (before endodontic treatment); 30 minutes after completion of endodontic treatment; and 4, 8, and 12 hours after the initial IN spray. Primary analysis was a repeated-measures analysis of variance. Results IN ketorolac alone or with endodontic treatment showed significantly better pain relief compared with IN placebo spray alone or with endodontic treatment at 30 minutes after the first or second intranasal dose and at 4 hours after the first intranasal dose (P = .03). Conclusions These results suggest that IN ketorolac may provide a novel and efficacious method for pain relief in endodontic pain patients.

The magnetic induced deposition of polydispersed aerosols composed of agglomerated superparamagnetic particles was measured with an in vitro model system and in the mouse trachea and deep lung for the purpose of investigating the potential of site specific respiratory drug delivery. Oleic acid coated superparamagnetic particles were prepared and characterized by TEM, induced magnetic moment, and iron content. The particles were dispersed in cyclohexane, aerosolized with an ultrasonic atomizer and dried by sequential reflux and charcoal columns. The fraction of iron deposited on glass tubes increased with particle size and decreasing flow rate. High deposition occurred with a small diameter tube, but the deposition fraction was largely independent of tube size at larger diameters. Results from computational fluid dynamics qualitatively agreed with the experimental results. Enhanced deposition was observed in the mouse lung but not in the trachea consistent with the analysis of the aerodynamic time allowed for deposition and required magnetic deposition time.

Lipid matrices containing dispersed superparamagnetic iron oxide (SPIO) particles were investigated as a magnetic field-responsive drug delivery system. Lipid matrices were prepared by combining myristyl alcohol, fatty acid coated SPIO particles, and umbelliferone (UMB). With placement of the matrices into the release medium, initial UMB release was fast but fell to zero indicating a burst effect. With application of an alternating magnetic field, additional UMB was released. The rate and extent of magnetic field-stimulated release increased with UMB load but not SPIO content. Differences between oleic and myristic acid coated SPIO appeared to be a result of phase separation. UMB release coincided with matrix melting, which can be controlled by the SPIO content and external magnetic field as shown by theoretical analysis. While significant technological issues remain, the foundation for developing magnetic field-stimulated drug delivery systems has been established.

The deposition of magnetic particles was examined for the possibility of further enhancing the selectivity of inhalation drug administration for the treatment of lung cancer.Superparamagnetic magnetite nanoparticles were prepared and ultrasonically atomized, dried, and passed through glass tubes in the presence and absence of a wedge-shaped permanent magnet. The change in the outlet aerosol size distribution due to magnetic deposition under various well-defined aerodynamic conditions and a measured magnetic field was determined by an aerodynamic particle sizer. In addition, computational fluid dynamics (CFD) simulations of magnetic aerosol transport and deposition were conducted.The deposition fraction increased nearly linearly with particle diameter and was greater with lower air flow rates. The effect of tube diameter was complicated but well described by CFD simulations, as was the effect of particle size and air flow rate.The descriptive power of CFD simulations was demonstrated in the in vitro deposition of magnetic aerosol particles. This suggests that CFD simulations can potentially be used in future studies to design systems for selective drug delivery in vivo as a function of magnetic properties, aerosol characteristics, and respiratory physiology.

The purpose of this study was to explore the properties of coated superparamagnetic nanoparticles (SPNs) specifically for their use in thermal-responsive drug delivery systems. Coated, magnetite SPNs were prepared and dispersed in cyclohexane or cetyl alcohol, a solid lipid at the physiological temperatures of 37 °C. The induced temperature change as a function of SPN concentration and external magnetic field and frequency was consistent with theoretical predictions. SPNs dispersed in a solid lipid matrix underwent heating and the associated melting occurred at a temperature suitable for a thermal-responsive drug delivery system.

Sodium cholesteryl carbonate ester (SCC) was synthesized, and its phase behavior was studied. The chemical structure was assessed by solid-state infrared spectroscopy based on vibration analysis. The wave number at 1705 and 1276 cm−1corresponds to a carbonyl carbonate and O–C–O stretching of SCC, respectively. Molecular structure of SCC was further investigated with1H and13C NMR spectroscopy. The chemical shift, for the carbonyl carbonate resonance appeared at 155.5 ppm. A molecular mass of SCC was atm/zof 452. Differential scanning calorimetry (DSC), video-enhanced microscopy (VEM) together with polarized light microscopy, and small-angle x-ray scattering (SAXS) were used to characterize the phase behavior as a function of temperature of SCC. Liquid crystalline phase was formed with SCC. Based on the thermal properties and x-ray diffraction, it appears that SCC forms a structure analogous to the type II monolayer structure observed with cholesterol esters.

Solid lipid particles were evaluated for their potential as a thermo-activated drug delivery system. Submicron-sized diphenylhexatriene (DPH)/myristyl alcohol particles were produced by an atomization/drying process and the release rate of DPH into sodium dodecyl sulfate (SDS) micellar solutions was measured. The results showed that the presence of micelles and thermal activation was necessary and sufficient for the release of DPH. The initial rate of DPH release was linear for most systems. However, nonlinearity was noted at low micelle concentrations where the rate decreased with time due to the loss of sink conditions with the rising concentration of DPH in the micellar solution. Also at high micelle and particle concentrations, the rate increased with time, which may be due to a loss of particle integrity. Analyzing the data from a design study, the release rate was found to be linearly proportional to particle concentration. In contrast, the rate of release increased with micelle concentration in a nonlinear manner and appeared to approach a plateau at high micelle concentrations. The decrease in rate as the micelle concentration increased is suggestive of a saturable process and may involve a collision-based mechanism.

Background Topical delivery of nebulized antibiotics to the paranasal sinuses has been shown to improve clinical outcomes in patients with chronic sinus disease after functional endoscopic sinus surgery (FESS). The most efficient method for delivering nebulized particles to the sinuses, however, has not been established. This study investigates how the size of nebulized particles influences the efficiency of deposition in the maxillary sinus of human cadavers after FESS. Methods Endoscopic maxillary antrostomy was performed on eight sides in four cadavers. Each cadaver's nasal vault was nebulized with technetium99m-labeled sulfur colloid particles of three size ranges. Anterior-posterior and left lateral static gamma-camera images of the head were captured with an acquisition cutoff limit of 30,000 gamma-count. Regions of interest were defined for the left and right maxillary sinus and gamma-photon counts were recorded. Analysis of variance (ANOVA) for repeated measures and paired t-test were used to determine statistical significance. Results Mean diameter of particles generated was 6, 0.99, and 0.67 μm. There was a statistically significant difference in deposition between the largest particle size and the two smaller sizes, with a mean gamma-photon count of 254 for 6-μm particles versus 811 for 0.99-μm particles and 835 for 0.67-μm particles (ANOVA, p = 0.002). Conclusion Particles in the 0.67- to 0.99-μm range had improved efficiency of deposition in the maxillary sinus compared with larger particles after maxillary antrostomy. Larger particles appeared to deposit directly in the nasal vault while smaller particles were more likely to reach the maxillary sinus.

The output and particle size distribution of several series of aqueous solutions were measured to define quantitatively the practical limits of the solution properties acceptable for aerosol production by the aeroneb micropump nebulizer. Aerosol output measurements were made gravimetrically and the particle size distributions were obtained by laser diffractometry. Solution properties were obtained from the literature by interpolation of the best-fit curve of the property plotted as a function of composition. For nonionic solutes, addition of sodium chloride dramatically increased the output rate and also decreased the droplet size at low solute concentrations. Increasing viscosity also caused a significant decrease in output. Cesium chloride displayed increased output rate with concentration due to the rising density. Based on calculations with the number of apertures and oscillatory frequency, low output rates appeared to be a consequence of apertures failing to produce a droplet with each oscillation. Overall, ionic strength, density, surface tension, and viscosity affected the output rate in a manner that can be now empirically predicted.

Effective chemoprevention of lung cancer in high-risk patients through the administration of pharmacologic or nutritional agents is urgently needed. Aerosol inhalation can deliver chemopreventive agents directly to the respiratory tract to inhibit the tumorigenic process. In this study, polyphenon E (PolyE) and (-)-epigallocatechin-3-gallate (EGCG) were administered by aerosol delivery to A/J mice beginning 2 weeks after carcinogen treatment and continuing daily by inhalation throughout the remainder of the study (20 weeks). PolyE decreased tumor load by approximately 59%. However, EGCG, both at the same dose and at a higher dose, failed to inhibit lung carcinogenesis. These results indicate that aerosol delivery of PolyE, but not EGCG, may be a useful chemopreventive protocol in subjects at high risk for lung cancer.

Because of the noninvasive, locally selective potential of thermal energy, considerable effort has been focused on the use of an external, alternating magnetic field for conversion of magnetic work to heat with iron oxide nanoparticles. However, proper regulation of thermal energy remains a challenge because of the lack of feedback from the local temperature change to the external power supply. Here, we show development of smart magnetic nanoparticles composed of Fe and Si with intrinsically tunable heat generation capability. They were engineered to possess an adjustable magnetic transition temperature through tuning the exchange between Fe atoms by incorporation of silicon atoms. They show relatively high magnetic moment. Moreover, their biocompatibility was established in several cell lines. The nanoparticles were also combined with a thermosensitive polymer, which had the capability to release of molecules with a magnetic stimulus, thereby providing a platform for locally controlled, drug release.

Lung cancer continues to be the number one cause of cancer-related deaths in the USA. Early identification of the disease, availability of more effective drugs, and improved delivery of such drugs specifically to cancer cells are needed to decrease lung cancer-associated morbidity and mortality. The concept of image-guided drug delivery (IGDD), which envisions the utilization of imaging techniques for quantitative assessments of tumor-targeted drug delivery and therapeutic response, has the potential to make a significant impact in lung cancer. While the anatomic and physiological features of the lung pose distinct problems for imaging drug delivery, several new techniques are emerging that have the potential to overcome these problems. X-ray is a routinely used technique for diagnosing lung cancer; however, positron emission tomography (PET) and magnetic resonance imaging (MRI) are complementary approaches. PET- and MRI-based techniques (such as functional MRI) offer the possibility of imaging the delivery of specific molecules to cancer tissues in the lung. This paper reviews fundamentals of imaging with an emphasis on MRI and to some extent PET, since it will be argued that these techniques are the most promising for development in IGDD for lung cancer. Finally, key literature contributions will be highlighted, which exemplify the current successes in this area.

The inhalation delivery of 5-fluorouracil (5-FU) in lipid-coated nanoparticles (LNPs) to hamsters was evaluated to determine the feasibility for use in lung cancer chemotherapy. The inhaled dose, 30 mg LNPs/kg body weight (1.5 mg/kg 5-FU), was delivered over an 8-min interval. Fluorescein isothiocyanate dextran (FITC-dextran) was included within the LNPs to provide an estimate of the particle concentration. The concentration of FITC-dextran and total 5-FU (released and LNP-associated) was determined as a function of time in the lung, trachea, larynx, esophagus, and serum. Concentrations of 5-FU and FITC-dextran were initially high in the trachea, larynx, and esophagus, and lower in the lung. Within 24 h, greater than 99% of the LNPs were cleared from the respiratory tract and total 5-FU concentrations mirrored the LNP concentration. An eight-compartment pharmacokinetic model was used to describe the observed trends in concentrations of LNPs and total 5-FU and to estimate the released 5-FU concentration in the above tissues. From this analysis, effective local targeting as well as sustained efficacious concentrations of 5-FU in the expected tumor sites were demonstrated.

The release rate of 5-fluorouracil (5-FU) from liposomes, microspheres, and lipid-coated nanoparticles (LNPs) was determined by microdialysis to investigate their use as a respirable delivery system for adjuvant (postsurgery) therapy of lung cancer. 5-FU was incorporated into liposomes using thin film hydration and into microspheres and LNPs by spray drying. Primary particle size distributions were measured by dynamic light scattering. Liposomes released 5-FU in 4-10 h (k(1) = 0.44-2.31/h, first-order release model). Extruded vesicles with diameters less than one micron released 5-FU more quickly than nonextruded vesicles. With poly-(lactide) (PLA) and Poly-(lactide-co-glycolide) (PLGA) microspheres, slower release rates were observed (k(1) = 0.067-0.202/h). Increasing the lactide:glycolide ratio (50:50-100:0) resulted in a progressive decrease in the release rate of 5-FU. poly-(lactide-co-caprolactone) (PLCL) microspheres released 5-FU more rapidly compared to PLGA systems (k(1) = 0.254-0.259/h). LNPs formulated with polymeric core excipients had lower release rates compared to monomeric excipients (k(1) = 0.043-0.105/h vs. k(1) = 0.192-0.345/h). Changing the lipid chain length of the shell lipid components had a relatively minor effect (k(1) = 0.043-0.129/h). Overall, these systems yielded a wide range of delivery durations that may be suitable for use as an inhalation delivery system for adjuvant therapy of lung cancer.

The release rate of 5-fluorouracil (5-FU) from lipid-coated nanoparticles (LNPs) was determined to develop a respirable delivery system for use as adjuvant (postsurgery) therapy for lung cancer. LNPs were prepared by spray drying, and the in vitro release was measured by microdialysis. The composition of the core and shell affected the release rate. Increasing the core diameter at constant shell thickness and increasing shell thickness at constant core diameter reduced the release rate, suggesting that the lipid shell is the rate limiting step for the release of 5-FU. A model consisting of a sequential zero-order/first-order dependence on time from polydispersed cores within polydispersed shells was developed to describe the release. Based on studies of the effect of geometry of the layered particles, the optimal formulation was identified as a 600-nm diameter 5-FU/poly-(glutamic acid) core with a 200-nm thick tripalmitin/cetyl alcohol shell. This system is readily aerosolized by ultrasonic atomization, which did not change the release properties. Preliminary instillation and inhalation delivery studies to the hamster resulted in lung levels of the particles and 5-FU that were near the desired values. Through this effort, a sustained-release, respirable delivery system for adjuvant therapy of lung cancer in humans may ultimately be realized.

The purpose of this study was to examine the feasibility of the microdialysis sampling technique as a method to precisely and conveniently measure drug release from microcarrier systems such as liposomes and microspheres. Release of 5-fluorouracil (5-FU) from liposomes and microspheres was evaluated in vitro using microdialysis. Retrodialysis calibration using 5-chlorouracil (5-CU) was performed in conjunction with on-line HPLC analysis. At a microdialysis perfusate flow rate of 0.5 muL/min, concurrent 5-FU gain and 5-CU loss ranged from 72% to 75%, while concurrent 5-FU loss and 5-CU ranged from 69% to 71%. After calibration, simultaneous 5-FU release profiles were obtained by continuous microdialysis and discrete equilibrium dialysis sampling using a side-by-side diffusion apparatus. Release rates were characterized by a first-order release model. The release rate constants for a representative liposomal formulation were 0.30 and 1.85/h by microdialysis in the acceptor and donor compartments, respectively, and 0.39/h by equilibrium dialysis in the acceptor compartment. The calculated release rate constant determined by equilibrium dialysis in the donor compartment (1.98/h) agrees with that determined by microdialysis (1.85/h) when the resistance of the equilibrium dialysis membrane with associated first-order rate constant of transfer of 0.42/h is taken into account. Release profiles of 5-FU from a number of different liposome and microsphere formulations were determined. The results indicate that a convenient and reproducible characterization of drug release from various liposome and microsphere formulations is readily obtainable by microdialysis.

The purpose of this work was to evaluate nanoindentation as a means to characterize the material properties of pharmaceutical solids. X-ray diffraction of potassium chloride and acetaminophen showed that samples prepared by cooling a melt to a crystalline sample as opposed to slow recrystallization had the same crystal structure. With analysis of the force-displacement curves, the KCl quenched samples had a hardness that was 10 times higher than the recrystallized KCl, while acetaminophen quenched samples were 25% harder than the recrystallized samples. The elastic moduli of the quenched samples were also much greater than that observed for the recrystallized samples. Although the elasticity was independent of load, the hardness increased with load for acetaminophen. With each sample, the flow at constant load increased with applied load. Etching patterns obtained by atomic force microscopy showed that the KCl quenched sample had a higher dislocation density than the recrystallized sample, although there was no evident difference in the acetaminophen samples. Overall, the differences in the observed sample properties may be related to the dislocation density. Thus, nanoindentation has been shown to be a sensitive method for determining a processed-induced change in the hardness, creep, and elasticity of KCl and acetaminophen.

The goal of our efforts has been to generate high aerosolized drug concentrations, so that the entire inhaled dose may be delivered to the animal in a relatively short time interval, typically one minute or less. In this report, we have examined the use of a reflux condenser coupled with an external heat source as a means to dry aerosol particles. The specific interest was to examine the parameters influencing the drying process and identify practical experimental conditions that would allow drying of aerosol particles. Aqueous solutions of cesium chloride were atomized with an ultrasonic driver, and the resulting cloud was subsequently passed through an externally heated column with an inner condenser cooled with flowing water. Increasing the airflow rate increased the output of total mass(water + CsCl) but decreased the particle transit efficiency. At a constant airflow rate, decreasing the wall-to-condenser temperature ratio led to a progressive increase in particle loss, but a maximum was observed in the water removal efficiency. While thermal diffusion and the thermophoretic effect may have an impact on the drying process, non-steady state conditions, convective currents, inertial impaction, turbulence, and changes in the condenser boundary layer are likely to have a role in particle drying. In spite of the numerous complexities, practical operating conditions were identified for efficient particle drying with high dry mass transit efficiency.

The force-displacement profiles of four well-characterized materials that represent both soft/hard and plastic/brittle materials have been obtained using a novel nanoindentation technique. Flat surfaces of acetaminophen, potassium chloride, sucrose, and sodium stearate were prepared by melting or recrystallization, and the melting points were measured. Topographic and the corresponding first derivative images were obtained both before and after indentation. The materials were indented using a 10 s loading time, followed by a 2 s hold, and ending with a 10 s unloading time thereby providing a unique force-displacement profile for each material. The loading profile of acetaminophen was discontinuous, whereas the profiles for the other three materials were smooth. The profiles were analyzed and the rank order of hardness was sucrose > acetaminophen > KCl > sodium stearate, which is consistent with the literature. The rank order of the stiffness, which is related to the modulus of elasticity, was sucrose > KCl > acetaminophen > sodium stearate. Given the flexibility and power of this approach, nanoindentation coupled with atomic force microscopy may be a useful means to characterize materials for evaluating tablet-processing conditions, perhaps at a preformulation stage.

The inhalation delivery of difluoromethylornithine (DFMO) was evaluated to determine its feasibility for use in lung cancer chemoprevention. Aerosol droplets of DFMO were produced, and the solvent was removed by a reflux drying column. Equivalent doses of DFMO (40 mg/kg) were given over 10 minutes by inhalation and by gavage, and the serum and tissue levels were determined. Inhalation of DFMO resulted in a peak lung concentration 80 times higher and area under the curve (AUC) that was 35 times higher than oral dosing. The estimated pharmacokinetic advantage based on serum and lung AUCs was 13 in favor of inhalation.

The association of hydrophobic, cationic drugs with lung surfactant was determined to assess the pharmacokinetic implications on drug disposition and retention in the lung.The distribution coefficients, K, were determined at 25 and 37 degrees in normal saline solution buffered at pH 7.4 for a series of structurally related, cationic drugs. Drugs were dispersed into lung surfactant, equilibrated, and then centrifuged to separate the aqueous phase from the surfactant pellet. Drug concentrations in the supernatant and pellet were determined following dilution using spectrophotometric assays. In addition, the apparent acid dissociation constant of quinacrine in the presence and absence of surfactant was determined by measuring the pH-dependent absorption spectra. The effect of stereochemistry on the distribution of drugs into surfactant was examined with (R)- and (S)-propranolol.The mole fraction distribution coefficients for amitriptyline, promethazine, promazine, ethopropazine, imipramine, R-propranolol, and S-propranolol at 25 degrees C were 6,560 +/- 500, 28,400 +/- 1,500, 12,100 +/- 840, 5,480 +/- 330, 4,490 +/- 250, 8,680 +/- 260, 8,190 +/- 530, respectively. At 37 degrees C, the distribution coefficients were generally smaller indicating a significant exothermic heat of transfer for these solutes from aqueous solution to the lung surfactant. The pKa of quinacrine was 7.43 +/- 0.04 in aqueous solution and was shifted to 7.62 +/- 0.06 in the presence of lung surfactant. From this shift, the double layer potential for quinacrine-lung surfactant was estimated to be -0.012 V assuming a dielectric constant equivalent to that of water.Cationic drugs have very favorable distributions from an aqueous solution to the lipid phase of lung surfactant. The transfer process generally has both a large entropic and enthalpic contribution. The latter thermodynamic aspect may be related to the charge interaction between the solute and the negatively charged surfactant. Finally, no significant effect of stereochemistry was evident with the distribution of (R)- and (S)-propranolol.

The effects of aerosol budesonide and dietary myo-inositol on progression of benzo[alpha]pyrene (B[alpha]P) induced carcinogenesis were studied in A/J mouse lung. First, we determined when to intervene in the carcinogenesis process by exposing several animals to B[alpha]P at 100 and 150 mg/kg of body wt. Groups of these animals were necropsied from 1 to 36 weeks post-carcinogen. The presence of different categories of lung tumors was noted over the 36 week time period. Hyperplasia first appeared approximately 6 weeks post-carcinogen followed by adenoma at 9 weeks, then by carcinoma at 26 weeks. From this temporal sequence we determined we could test for effects of preventive agents on progression to hyperplasia by intervening at 3 weeks, for effects on progression to adenoma by intervening at 6 weeks and for effects on progression to carcinoma by intervention at 12 weeks. Intervention at 3 weeks post-carcinogen with aerosolized budesonide delayed both hyperplasia and adenoma formation. Once hyperplasia appeared in budesonide treated animals, however, it increased at the same rate as in control animals, indicating a delay in progression. Progression from adenoma to carcinoma was reduced when budesonide was given 12 weeks post-carcinogen. Dietary myo-inositol failed to suppress progression from adenoma to carcinoma when started 12 weeks post-carcinogen. In summary, budesonide is a chemopreventive agent that has inhibitory effects on B[alpha]P induced carcinogenesis of the lung in A/J mice at all stages of progression from hyperplasia formation to cancer.

Purpose. The solubilization of a number of steroids was determined in bile salt simple micelles and a bile salt/phospholipid micellar system to provide a better basis to predict the extent of drug solubilization in vivo. Methods. Excess solid drug was dispersed in taurodeoxycholate or mixed micelle solutions prepared with fixed mole ratios of taurocholate, taurodeoxycholate, taurochenodeoxycholate, glycodeoxycholate, glycocholate, and glycochenodeoxycholate with egg phosphatidylcholine. Drug concentrations were determined from the absorbance following centrifugation. Using NMR spectroscopy, the diffusivities of the simple and mixed micelles were 2 × 10-6 and 8 × 10-7 cm2/s, respectively. Results. From the change in the concentration of drug in solution with a change in the lipid concentration, the solubilization ratio (SR) was calculated. The SR and aqueous solubility were used to calculate the micelle/aqueous partition coefficients (Km/w). Km/w was correlated with octanol/water partition (Po/w) for the TDC and mixed micelle data sets with correlation lines of logKm/w = 0.74logPo/w + 1.55 (r2 = 0.91) and logKm/w = 0.61 logPo/w + 2.44 (r2 = 0.95), respectively. Conclusions. With such data, a refined, predictive relationship between the in vitro and the in vivo solubilization with additional information concerning the bile salt/lipid concentration in the human intestine appears possible.

Particle size is a key determinant of biological performance of sub-micron size delivery systems. Previous studies investigating the effect of particle size have primarily focused on well-dispersed nanoparticles. However, inorganic nanoparticles are prone to aggregation in biological environments. In our studies, we examined the consequence of aggregation on superparamagnetic iron oxide (SPIO) nanoparticle-induced magnetic hyperthermia. Here we show that the extent and mechanism of hyperthermia-induced cell kill is highly dependent on the aggregation state of SPIO nanoparticles. Well-dispersed nanoparticles induced apoptosis, similar to that observed with conventional hyperthermia. Sub-micron size aggregates, on the other hand, induced temperature-dependent autophagy through generation of oxidative stress. Micron size aggregates caused rapid membrane damage, resulting in acute cell kill. Overall, this work highlights the potential for developing highly effective anticancer therapeutics through designed aggregation of nano delivery systems.

The interaction of sodium taurocholate/egg phosphatidylcholine (TC/PC) micelles with mucin was determined to investigate the exclusion of lipids by mucus in the absorption process. The distribution of TC/PC was assessed at two intermicellar and three phospholipid concentrations with isolated, rat intestinal mucin (RIM) by dialysis. The diffusion coefficients were measured by NMR spectroscopy. At high [PC], RIM had lower [PC] relative to the control, while the concentration of TC was largely independent of mucin concentration. The PC diffusion coefficients were reduced in the presence of RIM. The magnetization decay of TC was compared with simulations to provide estimates of the monomeric diffusivity and exchange rate constant. The rate constants increased with increasing micelle concentration, and the free TC diffusion coefficient was reduced in the presence of mucin. Mucin has an exclusive effect on TC/PC mixed micelles that has been quantitatively determined through the use of diffusion measurements of dialyzed samples.

The effect of a pulmonary surfactant extract from bovine lung, Survanta™, on the dissolution rate of aerosol particles of budesonide was determined. Aerosol particles of budesonide were generated from an ethanol solution, dried, and collected by a cascade impactor for characterization or by a liquid impinger for dissolution experiments. Powder x‐ray diffraction, differential scanning calorimetry, differential thermal analysis, and scanning electron microscopy were used to characterize the aerosol particles and starting material. No change in phase was detected, although the aerosol particles appeared to contain residual solvent. The dissolution rate of the aerosol particles in saline was low and variable. Survanta™ increased the extent of dissolution of budesonide in proportion to the added concentration, which was also verified by equilibrium solubilization studies. Survanta™ also increased rate of dissolution, in a manner similar to sodium dodecyl sulfate. Analysis of the concentration of budesonide following ultracentrifugation indicated that there is rapid equilibration of budesonide between the Survanta™ and aqueous phase. These results show that lung surfactant has the potential of enhancing the rate and extent of dissolution of drugs administered to the lung. © 2001 Wiley‐Liss, Inc. and the American Pharmaceutical Association J Pharm Sci 90: 98–104, 2001

Purpose. The permeability of rat intestinal mucin (RIM) to sodium taurocholate/egg phosphatidylcholine (TC/PC)-mixed micelles has been investigated.

Purpose. The purpose of this study was to determine the distribution and diffusion of sodium taurocholate-phospholipid micelles with mucin in order to provide the foundation for understanding the transport of ingested fat and poorly water-soluble drugs through the intestinal mucous layer.

The relative affinity of glucocorticosteroids for lung surfactant was determined for the purpose of identifying chemopreventive agents with a high therapeutic index for lung cancer. The aqueous solubility and the extent of solubilization in Survanta, a native extract of bovine lung, of budesonide, triamcinolone acetonide, dexamethasone, and flunisolide were determined as a function of temperature by a dialysis technique. The aqueous solubilites at 37 degrees C were 19.6, 35.8, 104 and 120 microg/ml for the above listed compounds, respectively. The temperature dependence of the solubilities was modest consistent with the hydrophobic properties of the steroids. The amount of drug in solution was significantly enhanced in the presence of Survanta with solubilization ratios of 0. 019, 0.023, 0.014, and 0.02 microg drug dissolved per microg of Survanta phospholipid, respectively. In addition, the extent of solubilization also generally increased with temperature, although the phase transition of the surfactant lipid appeared to complicate the functional relation between temperature and solubilization. These results show that there is enhanced solubilization of glucocortosteroids by lung surfactant which is secreted by the cancer susceptible type II cells of the lung.

Purpose. The purpose of this study was to analyze a diffusion dryer as a means to remove organic solvents from aerosol particles of poorly water soluble drugs.

Purpose: The spectral properties and ionization behavior of four retinoids: retinoic acid; MTTO; TTNN; and TTNPB, have been determined in aqueous solution to provide a better understanding of their aggregation behavior. Methods: The solubility and ultraviolet absorption spectrum were measured in solutions prepared by equilibration with solid and by dilution of an ethanol solution with buffer. The UV spectral shift was measured as a function of pH to quantify the extent of ionization in 5 and 150 mM NaCl. Fluorescence polarization measurements of TTNN were made. Results: The solubility of the triclinic form of retinoic acid was lower than that of the monoclinic indicating the former is the more stable polymorphic form. For the retinoids, the measured solubility increased with the number of phenyl rings. The wavelengths of maximum absorption and absorptivity of the non-ionized forms of the retinoids systematically changed with chemical structure provided that the preferred molecular conformation was considered. High apparent negative logarithms of the ionization constant, pKaobs, were observed indicating that the retinoids undergo self-association. The titration curves were fit to the Henderson–Hasselbalch relationship, and the pKaobs was dependent on the retinoid concentration in solution. The value of the pKa at low ionic strength was slightly larger that that obtained at high ionic strength. The fluorescent polarization suggested that the aggregation number changes with ionic strength and extent of ionization. Finally, the difference between the observed pKa and the pKa of model compounds was smaller for retinoic acid than for MTTO, TTNPB, and TTNN. Conclusions: All retinoids under study were self-associated giving rise to relatively high values of the apparent pKa. The properties of the aggregates are dependent on the concentration and ionization, and as such, the retinoids probably lack a well-defined aggregation number and critical micelle concentration. The nature of the aggregation for retinoic acid appears distinct from that of the arotinoids, MTTO, TTNPB, and TTNN.

Purpose. The aqueous solubility and the extent of solubilization and ionization constant in sodium taurodeoxycholate (NaTDC) solutions of a series of benzoic acid and aniline derivatives were measured as a basis to characterize and thereby help predict the nature of the interaction of drugs with bile aggregates. Methods. The aqueous solubility and the solubilization of two series of compounds, 4-alkyl benzoic acids and 4-alkyl anilines, was measured as a function of NaTDC in 0 and 150 mM NaCl. The ionization constants were determined in water and in 50 mM NaTDC at sodium chloride concentrations of 0, 75 and 150 mM by spectrophotometric titration. The diffusion coefficients of NaTDC and the solutes were measured by pulsed-field gradient spin echo NMR spectroscopy. Results. The aqueous solubilities decreased with increasing alkyl chain length in both series, and the aniline derivatives had larger solubilities than the benzoic acid derivatives. The number of moles of solute solubilized per mole of bile salt ranged from 0.17 to 0.31 for the benzoic acid derivatives and from 1.3 to 3.0 for the aniline derivatives. The pKa values of the benzoic acid derivatives in the presence of NaTDC were higher relative to the controls, and the difference in the pKa (ΔpKa,obs) increased with increasing chain length. With the aniline derivatives, the pKa values were also shifted to higher values in NaTDC relative to the control but only in the absence of salt. The presence of the solute caused a decrease in the diffusion coefficient of NaTDC, and the diffusion coefficients of the solutes decreased with increasing alkyl chain length. With the hexyl derivatives, the diffusion coefficient of the solute was smaller than the diffusion coefficient of the bile salt. The chemical shift of the protons attached to carbon 18 and 19 of the bile salt were decreased to a greater extent in the presence of the solutes than the protons attached to carbon 26. Conclusions. Both the solubilization and ionization behavior of solutes were affected by the presence of bile salt aggregates. The surface potential and effective polarity of NaTDC aggregates were found to be dependent on the alkyl chain length for these two homologous series of solutes. The solubilization ratio was largely independent of alkyl chain length, but the unitary partition coefficient was dependent on both the alkyl chain length as well as ionization state. The derivatives reduced the diffusivity of the micelles suggesting the formation of larger sized aggregates and the solutes (hexyl derivatives) appear to favor association with the larger sized aggregates. The phenyl ring of the solutes appears to be oriented parallel to the plane of the steroid frame with preferential positioning near the hydrophobic rings.

Cancer stem cells (CSCs) are a subpopulation of cancer cells that have stem cell-like properties and are thought to be responsible for tumor drug resistance and relapse. Therapies that can effectively eliminate CSCs will, therefore, likely inhibit tumor recurrence. The objective of our study was to determine the susceptibility of CSCs to magnetic hyperthermia, a treatment that utilizes superparamagnetic iron oxide nanoparticles placed in an alternating magnetic field to generate localized heat and achieve selective tumor cell kill. SPIO NPs having a magnetite core of 12 nm were used to induce magnetic hyperthermia in A549 and MDA-MB-231 tumor cells. Multiple assays for CSCs, including side population phenotype, aldehyde dehydrogenase expression, mammosphere formation, and in vivo xenotransplantation, indicated that magnetic hyperthermia reduced or, in some cases, eliminated the CSC subpopulation in treated cells. Interestingly, conventional hyperthermia, induced by subjecting cells to elevated temperature (46 °C) in a water bath, was not effective in eliminating CSCs. Our studies show that magnetic hyperthermia has pleiotropic effects, inducing acute necrosis in some cells while stimulating reactive oxygen species generation and slower cell kill in others. These results suggest the potential for lower rates of tumor recurrence after magnetic hyperthermia compared to conventional cancer therapies.

The platelet-derived growth factor (PDGF) signaling pathway has been found to be activated in human pulmonary arterial hypertension (PAH) and in animal models of the disease. Our study tested the hypothesis that a novel, nonselective inhaled PDGF receptor inhibitor, PK10453, would decrease pulmonary hypertension both in the rat monocrotaline (MCT) model and the rat MCT plus pneumonectomy (MCT+PN) model of PAH. PK10453, delivered by inhalation for 4 (D4)- and 8 (D8)-minute exposures 3 times a day for 2 weeks, decreased right ventricular systolic pressure (RVSP) in both the rat MCT and rat MCT+PN models: RVSP was 80.4 ± 2.6 mmHg in the vehicle MCT group (n = 6), 44.4 ± 5.8 mmHg in the D4 MCT group (n = 6), and 37.1 ± 4.5 mmHg in the D8 MCT group (n = 5; P < 0.001 vs. vehicle); RVSP was 75.7 ± 7.1 mmHg in the vehicle MCT+PN group (n = 9), 40.4 ± 2.7 mmHg in the D4 MCT+PN group (n = 10), and 43.0 ± 3.0 mmHg in the D8 MCT+PN group (n = 8; P < 0.001). In the rat MCT+PN model, continuous telemetry monitoring of pulmonary artery pressures also demonstrated that PK10453 prevented the progression of PAH. Imatinib given by inhalation was equally effective in the MCT model but was not effective in the MCT+PN model. Immunohistochemistry demonstrated increased activation of the PDGFβ receptor compared to the PDGFα receptor in neointimal and perivascular lesions found in the MCT+PN model. We show that imatinib is selective for the PDGFα receptor, whereas PK10453 has a lower half-maximal inhibitor concentration (IC50) for inhibition of kinase activity of both the PDGFα and PDGFβ receptors compared to imatinib. In conclusion, PK10453, when delivered by inhalation, significantly decreased the progression of PAH in the rat MCT and MCT+PN models. Nonselective inhibition of both the PDGFα and PDGFβ receptors may have a therapeutic advantage over selective PDGFα receptor inhibition in PAH.

Bile salt/phospholipid aggregates are present as concentrated, anionic colloids in the gallbladder. After mixing with dietary lipids, these aggregates diffuse through the hydrated, negatively charged glycoprotein network at the intestinal surface prior to absorption. Analysis of these complicated processes requires a characterization of the lipid aggregate as well as its diffusion. Moreover, the excluded-volume effects on the lipid aggregate diffusion need to be addressed. Thus, the diffusion coefficients of bile salt, phospholipid, and water were obtained from four series of lipid dilutions by a Fourier transform pulsed-field gradient spin-echo 1H NMR diffusion method. Dialysis experiments were performed to provide an independent estimate of the intermicellar concentration. Four lipid solutions with different total phospholipid/bile salt ratios were then diluted with bile salt solutions containing the approximate intermicellar concentration. With these four series of solutions, the isopotential specific ...