Rationale: Several extrapulmonary disorders have been linked to cigarette smoking. Smoking is reported to cause cystic fibrosis transmembrane conductance regulator (CFTR) dysfunction in the airway, and is also associated with pancreatitis, male infertility, and cachexia, features characteristic of cystic fibrosis and suggestive of an etiological role for CFTR. Objectives: To study the effect of cigarette smoke on extrapulmonary CFTR function. Methods: Demographics, spirometry, exercise tolerance, symptom questionnaires, CFTR genetics, and sweat chloride analysis were obtained in smokers with and without chronic obstructive pulmonary disease (COPD). CFTR activity was measured by nasal potential difference in mice and by Ussing chamber electrophysiology in vitro. Serum acrolein levels were estimated with mass spectroscopy. Measurements and Main Results: Healthy smokers (29.45 6 13.90 mEq), smokers with COPD (31.89 6 13.9 mEq), and former smokers with COPD (25.07 6 10.92 mEq) had elevated sweat chloride levels compared with normal control subjects (14.5 6 7.77 mEq), indicating reduced CFTR activity in a nonrespiratory organ. Intestinal current measurements also demonstrated a 65% decrease in CFTR function in smokers compared with never smokers. CFTR activity was decreased by 68% in normal human bronchial epithelial cells exposed to plasma from smokers, suggesting that one or more circulating agents could confer CFTR dysfunction. Cigarette smokeexposed mice had decreased CFTR activity in intestinal epithelium (84.3 and 45%, after 5 and 17 wk, respectively). Acrolein, a component of cigarette smoke, was higher in smokers, blocked CFTR by inhibiting channel gating, and was attenuated by antioxidant N-acetylcysteine, a known scavenger of acrolein. Conclusions: Smoking causes systemic CFTR dysfunction. Acrolein present in cigarette smoke mediates CFTR defects in extrapulmonary tissues in smokers.

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We assessed the safety and efficacy of combined intravenous and aerosolized antioxidant administration to attenuate chlorine gasinduced airway alterations when administered after exposure. Adult male Sprague-Dawley rats were exposed to air or 400 parts per million (ppm) chlorine (a concentration likely to be encountered in the vicinity of industrial accidents) in environmental chambers for 30 minutes, and returned to room air, and they then received a single intravenous injection of ascorbic acid and deferoxamine or saline. At 1 hour and 15 hours after chlorine exposure, the rats were treated with aerosolized ascorbate and deferoxamine or vehicle. Lung antioxidant profiles, plasma ascorbate concentrations, airway morphology, and airway reactivity were evaluated at 24 hours and 7 days after chlorine exposure. At 24 hours after exposure, chlorine-exposed rats had significantly lower pulmonary ascorbate and reduced glutathione concentrations. Treatment with antioxidants restored depleted ascorbate in lungs and plasma. At 7 days after exposure, in chlorineexposed, vehicle-treated rats, the thickness of the proximal airways was 60% greater than in control rats, with twice the amount of mucosubstances. Airway resistance in response to methacholine challenge was also significantly elevated. Combined treatment with intravenous and aerosolized antioxidants restored airway morphology, the amount of airway mucosubstances, and airway reactivity to control levels by 7 days after chlorine exposure. Our results demonstrate for the first time, to the best of our knowledge, that severe injury to major airways in rats exposed to chlorine, as characterized by epithelial hyperplasia, mucus accumulation, and airway hyperreactivity, can be reversed in a safe and efficacious manner by the postexposure administration of ascorbate and deferoxamine.Keywords: epithelial injury; epithelial repair; mucosubstances; ascorbate; deferoxamine; aerosol Chlorine is essential to global industry and to global public health. According to the World Chlorine Council (http://www. worldchlorine.org), 14.4 million metric tons were produced in North America in 2008, and 62.8 million metric tons were produced globally. Water treatment makes up only 5% of the world's use of chlorine. The majority of chlorine is used in the production of plastics such as polyvinyl chloride (1). It is also used as a bleaching agent for pulp and paper production, as feedstock in the production of chlorinated solvents used in metalworking, in dry cleaning, in electronics, and in pharmaceutical production (1-6).Only 20 American states contain facilities that produce chlorine, but every American state has facilities that use chlorine. This results in the shipping of chlorine by railcar, and increases the potential for large-scale accidents. According to the Environmental Protection Agency, chlorine gas was related to 518 serious accidents over a 5-year period during the 1990s (1). Multiple-casualty exposures to chlorine have resulted from industrial accidents involving chlorine ta...

To create an allergy model in the dog, allergic Beagles with high levels of serum immunoglobulin E (IgE) and eosinophilia were bred; resulting puppies were sensitized to ragweed by intraperitoneal (IP) injection within 24 hours of birth through 22 weeks of age. At least 50% of the puppies developed high levels of serum IgE and eosinophilia. As young adults, 6 of these dogs, and 6 control age-matched, nonallergic, nonimmunized dogs were exposed by inhalation to ragweed twice at 13-day intervals, and a third time 45 days later. Total and ragweed-specific serum IgE and ragweed-specific serum IgG were increased significantly in allergic dogs relative to baseline. Allergic dogs had significantly greater levels of antibody specific for ragweed, as well as higher eosinophil counts in the bronchoalveolar lavage fluid, compared to nonallergic dogs. Airway reactivity to histamine in allergic, but not nonallergic dogs, increased significantly after aerosol exposure to ragweed. After a third exposure to ragweed, airway responses to histamine were elevated in the allergic dogs and remained high for at least 5 months. These results demonstrate the potential of the allergic dog model for investigating the underlying pulmonary immune mechanisms and therapeutic treatment of allergic asthma.

MethodDust storm aerosol concentrations and particle size distributions have been measured in many countries. The mean aerosol concentration of a moderate dust storm is 0.040 mg/L, and the particle size is less than or equal to 2.5 m (Chan, 2002;Selinus, 2005). Therefore, the test plenum target aerosol concentration was 0.40 mg/L and the target particle size was 1.0 to 2.5 m. Although this particle size range is larger than the actual geometric diameter of viral particles (approximately 0.02 to 0.2 m), it should be noted that droplet nuclei generated during a sneeze range in size from 0.5 to 12 m and contain many viral particles (3M Technical Data Bulletin #174, 2004).Adults breathe at a rate of approximately 7.5 L/min while resting and 13 to 25 L/min during light exercise (Adams, 1993). The mannequin filter sampler volumetric flow rate was set at 8.75 L/min to simulate a near resting state respiratory ventilation rate. Efficiency data collected under these conditions represented a best-case scenario for protection; that is, protection would presumably be less and inhaled dose greater at light exercise ventilatory flow rates. The reference sampler volumetric flow rate was set at 1.72 L/min. Mannequin and reference filter sampler flow rates were metered with custom-designed critical orifi.A rectangular plenum with a volume of 147.5 L was used to test the masks. Aerosols were generated using an IV HEART™ nebulizer operated at 40 psig and 12.7 L/min. At this volumetric flow rate, the theoretical time to fill the test plenum with aerosol was 11.6 minutes. Therefore, the nebulizer was run for 12 minutes prior to filter samples being collected. All volumetric flow rates were calibrated using a DryCal DC-Lite (BIOS International, Butler, NJ) primary flow calibration device. A schematic of the face mask test system is presented in Figure 1. ProcedureA Styrofoam™ mannequin head was fitted with a sample probe. Face masks and a N95 respirator were placed on the mannequin head and positioned in the test plenum. Pictures of the face masks on the mannequin head are presented in Figures 2-5. A reference sample probe was positioned next to the mannequin head, and filter samplers were connected to the mannequin head and reference sample probes. The nebulizer was filled with approximately 20 mL of 0.045% saline, connected to the compressed air source and placed in the test plenum.The nebulizer was actuated and allowed to run for 12 minutes. Mannequin and reference filter samplers were actuated simultaneously, and 30-minute aerosol samples were collected. Initial and final filter pressure differentials were recorded from magnehelic pressure gages. Pres-

Models of emphysema produced by exposing animals to cigarette smoke (CS) have potential for use in testing treatments of this disease. To better characterize development of emphysema in an animal model, male and female mice of the B6C3F1 and A/J strains were exposed to CS at 250 mg total particulate material (TPM)/m3 for 15 weeks. Emphysema was evident in both strains of mice to differing degrees of severity. The CS-induced increase in the mean linear intercept (normalized to BW) of A/J mice was 51% greater than the control value, while CS-exposed B6C3F1 had an increase of 38% in this morphometric measurement of alveolar air space enlargement. In separate experiments, female B6C3F1 mice and male A/J mice were exposed to CS for 32 weeks and 15 weeks, respectively, and were then used to test the efficacy of all trans-retinoic acid (ATRA) treatments to ameliorate emphysema lesions. Following CS exposure, the B6C3F1 mice were treated once daily for 14 days in a 3-week period by nose-only inhalation exposure to aerosols of 180 or 1,800 mg-minutes ATRA/m3. The A/J mice were treated once daily, 4 days/week, for three weeks by either intraperitoneal injection of ATRA (0.5 or 2.5 mg/kg) or inhalation exposure to ATRA (3,600 or 18,000 mg-minutes/m3). Neither the injections nor inhalation exposures of ATRA in either strain of mouse caused reversal of the emphysema. In summary, CS-induced emphysema was more severe in A/J mice than in B6C3F1 mice. Treatment with ATRA did not reverse emphysema in either strain of CS-exposed mice.

A past study demonstrated that all-trans-retinoic acid (ATRA) treatment by intraperitoneal injection in a rat model of elastase-induced emphysema caused tissue regeneration as evidenced by a decrease in alveolar size and lung volume and an increase in alveolar number. We postulated that treatment with this retinoid by nose-only inhalation exposure would be a more efficient means of targeting damaged lung tissue. Emphysema was induced in male Fischer 344 rats by intratracheal instillation of pancreatic elastase (0.5 IU/g body weight). Four weeks after elastase instillation, animals were treated once daily, 4 days/week, for 3 weeks by exposing them nose-only to aerosolized ATRA (target concentration-time of 3000 or 15,000 mg-min/m3) or by injecting them intraperitoneally with ATRA in cottonseed oil (0.5 or 2.5 mg/kg). Based on estimates of particle deposition in the respiratory tract, inhalation doses were chosen to be consistent with injected doses. Lungs were fixed by inflation with formalin (constant pressure for 6 hours followed by >48 hours of immersion) and were embedded in paraffin. Sections were evaluated by histopathology and stereology. Inhalation exposure to ATRA at both aerosol concentrations caused significant elevations of ATRA in the lung, whereas only the high-dose injection treatment was associated with an elevation of lung ATRA. The mean ATRA concentration from lungs of rats in the high-dose inhalation exposure groups as measured by liquid chromatography--mass spectrometry was approximately 12-fold greater than that of high-dose injection-treated rats. Elastase instillation caused increased lung volumes, irregular alveolar air space enlargement, and fragmentation and attenuation of alveolar septa. Neither inhaled nor injected ATRA reduced the enlarged lung volumes associated with this emphysema model. Stereology demonstrated that alveolar air space enlargement in ATRA-treated rats was similar to that in sham-treated emphysematous animals. Thus, while inhalation treatment caused greater levels of the drug in lung tissue in comparison to that of injection-treated animals, treatment with ATRA by either route of administration did not cause a reversal of lung tissue damage in this model of elastase-induced emphysema.

The results of the present study, using the conscious beagle dog, demonstrate that inhaled insulin (INH; Exubera) provides better glycemic control during an intraportal glucose load than identical insulin levels induced by insulin (Humulin) infusion into the inferior vena cava (IVC). In the INH group (n ‫؍‬ 13), portal glucose infusion caused arterial plasma glucose to rise transiently (152 ؎ 9 mg/dl), before it returned to baseline (65 min) for the next 2 h. Net hepatic glucose uptake was minimal, whereas nonhepatic uptake rose to 12.5 ؎ 0.5 mg ⅐ kg ؊1 ⅐ min ؊1 (65 min). In the IVC group (n ‫؍‬ 9), arterial glucose rose rapidly (172 ؎ 6 mg/dl) and transiently fell to 135 ؎ 13 mg/dl (65 min) before returning to 165 ؎ 15 mg/dl (125 min). Plasma glucose excursions and hepatic glucose uptake were much greater in the IVC group, whereas nonhepatic uptake was markedly less (8.6 ؎ 0.9 mg ⅐ kg ؊1 ⅐ min ؊1 ; 65 min). Insulin kinetics and areas under the curve were identical in both groups. These data suggest that inhalation of Exubera results in a unique action on nonhepatic glucose clearance. Diabetes 54:1164 -1170, 2005 I nsulin delivered by inhalation is an alternative to injected insulin for patients with diabetes. Clinical studies comparing subcutaneous administration of insulin (Humulin) with insulin inhalation (Exubera) suggest that there is a unique glucose-lowering effect associated with inhaled insulin (1). Previous studies in dogs have shown that inhaled insulin has a higher and earlier peak of arterial insulin that clears more rapidly than subcutaneous insulin. Despite the similar arterial and hepatic insulin areas under the curve (AUCs) in the two groups, a 20% greater glucose infusion rate was required to maintain euglycemia in the inhaled insulin group relative to the subcutaneous insulin group (2). This finding was consistent with the unique glucose-lowering effect of inhaled insulin seen in human studies.The difference in glycemic control resulting from inhalation verses subcutaneous administration of insulin could be explained by the different pharmacokinetic insulin profiles resulting from the two routes of administration, or by some unique aspect of insulin delivery by inhalation. To distinguish between these possibilities, we delivered insulin by inhalation (Exubera) or through infusion (Humulin) into the inferior vena cava (IVC) to create an identical pharmacokinetic profile to that resulting from insulin inhalation. We then examined the effects of both routes of insulin delivery on glucose disposal during a simulated oral glucose load. To further clarify the mechanism by which glucose disposal might be enhanced, we compared the role of the liver and nonhepatic tissues in glucose uptake. RESEARCH DESIGN AND METHODSExperiments were conducted on 22 healthy, conscious, 18-h-fasted, male beagle dogs (weight 8 -10 kg). Before the study, they were fed a standard diet once a day, and water was provided ad libitum. The surgical facility met the standards published by the American Association for the Accredi...