Clinical characteristics of asthmatic patients prescribed various beta-agonist metered-dose inhalers at Yamagata University Hospital.

To determine the prescription characteristics of beta-agonist metered-dose inhalers (MDI), we retrospectively investigated all prescriptions containing one of five types of beta-agonist MDIs available at Yamagata University Hospital in 1997, as well as patients' characteristics. The total number of asthmatic patients was 225 (age, 11-79, mean, 47.2) in 1997. Fenoterol MDI was prescribed to patients who visited the hospital at regular periods and had more severe asthma. Isoprenaline MDI also was not prescribed for first-time patients. Patients who were prescribed tulobuterol MDI had mild or moderate asthma and some of them were only occasional or first-time visitors. Salbutamol and procaterol MDIs were also prescribed for first-time patients; however, tulobuterol MDI was the most frequently prescribed for first-time patients. Patients prescribed fenoterol and isoprenaline MDIs had adequate knowledge of proper asthma management, because sufficient information had been provided about the use of MDIs in the past. Patients prescribed tulobuterol MDI should be provided with detailed instructions because they had little knowledge of handling MDIs and self-management of asthma as many of them were first or intermittent visitors. Patients prescribed salbutamol or procaterol MDIs should be evaluated regarding their past medications and some of them should be instructed regarding the use of the MDI. Although these clinical aspects might be applicable only to our hospital, the same or other prescription patterns will be found in other hospitals and/or by other physicians. Adequate instructions to individual patients who are prescribed a particular beta-agonist MDI should be provided by the medical staff, especially to outpatients, to reduce hospitalization and death from asthma.

Histamine release induced by antimicrobial agents and effects of antimicrobial agents on vancomycin-induced histamine release from rat peritoneal mast cells.

Vancomycin and certain fungicides may cause anaphylactoid reactions. We investigated the effects of vancomycin, miconazole and fluconazole on histamine release in rat peritoneal mast cells. Vancomycin and miconazole provoked histamine release in a dose-dependent manner. In contrast, fluconazole did not provoke histamine release at concentrations of 3 x 10(-6)-3 x 10(-3) M. Vancomycin is efficacious in the treatment of gram-positive bacterial infections; patients presenting themselves with mixed infections require concomitant therapy with a second antimicrobial agent. We investigated the effect of fosfomycin sodium, cilastatin sodium or fluconazole on vancomycin-induced histamine release. Fosfomycin sodium inhibited vancomycin-induced histamine release but neither cilastatin sodium nor fluconazole inhibited it in the mole ratios of daily doses used in humans. These results suggest that vancomycin and miconazole provoke histamine release in rat mast cells, but that fluconazole probably does not, while fosfomycin sodium may inhibit vancomycin-induced histamine release.

Development of a new method for simultaneously evaluating mucociliary clearance and pulmonary epithelial permeability in rabbit experiments by means of 18FDG, three-dimensional positron emission tomography and rectilinear scan.

We tried to simultaneously obtain the elimination constant of mucociliary clearance and the pulmonary epithelial permeability constant after inhalation of 2-[18F]fluoro-2-deoxy-D-glucose (18FDG) solution by carrying out whole lung positron emission tomography and a rectilinear scan in rabbit experiments. The elimination constant of pulmonary epithelial permeability was obtained from the decrease in the amount of the radioactivity with time in the region of interest (ROI) confined to the lungs, trachea and tracheal cannula in the rectilinear scan. The total elimination constant of the radioactivity in the lungs was obtained from the ROI confined to the lungs in the tomography. The mucociliary clearance rate constant in the lungs was then obtained after subtracting the elimination constant of the pulmonary epithelial permeability from the total elimination constant of the 18FDG in the lungs. The mucociliary clearance constant in the trachea was calculated from the residual radioactivity in the trachea and the mucociliary clearance constant in the lungs. The mean pulmonary epithelial permeability constant was 0.0020% min(-1) obtained from the rectilinear scan. The mean mucociliary clearance constants of the lungs and the trachea were 0.0006 and 0.025% min(-1), respectively. These results indicated that the pulmonary epithelial permeability and mucociliary clearance could be evaluated simultaneously with 18FDG by using three-dimensional positron emission tomography and a rectilinear scan.

Biodistribution study of murine monoclonal anti-GD3 antibody in nude mice bearing human melanoma xenografts for development of immunoscintigraphy.

Reactivity of the monoclonal antibody with the tumor markers is known to be different between cultured cells in vitro and transplanted tumors in vivo. The monoclonal antibody should be investigated regarding its specific accumulation in tumor-bearing mice for immunodetection or immunotherapy. We studied the biodistribution of radiolabeled monoclonal anti-GD3 antibody (IgM) in normal mice and nude mice bearing human melanoma xenografts. Tissue-to-blood distribution ratios of the antibody in the liver, spleen and kidney increased with time in both normal and melanoma-transplanted mice, but no significant changes were observed in other normal tissues up to 5 days after injection. Specific accumulation of the monoclonal anti-GD3 antibody in the grafted human melanoma (HMV-II) was observed 4 and 5 days after injection. On the other hand, no specific accumulation of standard murine IgM in the tissue of HMV-II was observed in mice bearing the HMV-II xenograft 5 days after injection. Because the tissue-to-blood ratio of the distribution in the tissue of HMV-II became larger than that of other tissues 4 and 5 days after administration, 4 days after the administration of the monoclonal anti-GD3 antibody were required for immunoscintigraphy. Accumulation of the monoclonal anti-GD3 antibody in other human melanomas (HMV-I, HMY-1 and SK-MEL188) inoculated into mice was also observed 4 days after the antibody administration. The monoclonal anti-GD3 antibody used in this study would be useful in immunodetection or immunotherapy.

Deposition and clearance of inhaled 18FDG powder in patients with chronic obstructive pulmonary disease.

As freon is limited in its use as a generator for aerosol inhalation, powder particles are used as an alternative for inhalation therapy. The pulmonary deposition and clearance of inhaled powder particles was studied by positron emission tomography (PET) in ten patients with chronic obstructive pulmonary disease (COPD) and in five normal controls. The powder, 5 microm in mean diameter, was water soluble and labelled with 2-deoxy-2-[18F]-fluoro-D-glucose (18FDG). Powder inhalation was done with single deep inspiration from residual volume to total lung capacity. The initial deposition ratio in the right or left lung field to total inhaled dose, measured by an anteroposterior rectilinear scan, did not differ between normals and COPD patients. Ratios of radioactivity detected within the central and peripheral regions (the central to peripheral ratio) measured by the PET scan was not significantly different between COPD patients (4.8+/-2.6, mean+/-SD) and normals (2.6+/-0.8, mean+/-SD). However, the regional powder deposition in peripheral lung fields measured by the PET scan was significantly more uneven in COPD patients than in normal patients. The clearance rate of 18FDG, defined as the retention ratio of 18FDG activity to the initially deposited 18FDG at 60 and 120 min after inhalation, in the trachea, large bronchi or peripheral lung fields measured by tomographic scan showed a wider variation in COPD patients than in normals. To conclude, inhaled powder tended to be deposited more centrally and was distributed more unevenly in the peripheral lung in chronic obstructive pulmonary disease patients than in normals. This could be a limitation of powder inhalation used for therapy in chronic obstructive pulmonary disease patients.

[Investigation of the effects of puff times and spacer devices on the distribution pattern of a Becotide 100 inhaler in the twin impinger].

In order to investigate the dispersion pattern and compare the effect of spacer devices (Volumatic, Volumatic Soft and InspirEase), the Becotide 100 inhaler actuating 100 micrograms of beclomethasone dipropionate (BDP) per shot was studied with respect to its distribution ratio in a twin impinger and also compared with that of the Becotide 50 inhaler. The distribution patterns of BDP during each stage of the twin impinger were not affected by the puff times of the Becotide 100 inhaler. No significant difference was observed when the distribution ratios between the Becotide 50 inhaler and Becotide 100 inhaler were compared. All of these three spacer devices reduced the deposition ratio in stage I which is assumed to be the oral cavity and oropharynx. As the puff times of the Becotide 100 inhaler increased, the BDP deposited in the spacer increased and that of stage II assumed to be the lung decreased. These results suggested that the inhalation volume of BDP would depend on the puff times during clinical use and the inhalation volume of BDP for the Becotide 100 inhaler would be double that of the Becotide 50 inhaler. The spacer devices could decrease the local adverse effects caused by deposited BDP in the oral cavity. When using large spacer devices, one or two puff times into the spacer would be recommended for the best clinical effectiveness. The Volumatic could be expected to produce a superior BDP inhalation volume by reducing the deposition in the oral cavity in the 3 spacer devices examined in this study.

[Comparison by twin impinger of the distribution patterns of two beclomethasone dipropionate metered dose inhalers and of two spacer devices].

A Twin Impinger was used to assess the influence of the number of puffs on the distribution patterns of two commercially available beclomethasone dipropionate (BDP) metered dose inhalers, and the effects of two kinds of spacer devices were also compared. No difference was observed in the distribution pattern up to the 10th puff between the two metered dose inhalers. Clinically, the amount of BDP inhaled into the lungs is said to depend on the number of puffs taken, but in this study, there was a significant difference in the distribution pattern between these 2 inhalers. In the stage assumed to represent the oropharynx site, the distribution ratio of BDP was greater after administration with an Aldecin than with a Becotide inhaler. In contrast, in the stage assumed to represent the lung, the distribution ratio of BDP was greater with the Becotide inhaler. This result suggests that the amount of BDP inhaled in the lungs may vary between these two preparations. The effects of spacer devices were also investigated in 4 puffs. These spacer devices caused the distribution ratio in the stage assumed to represent the oropharynx to decrease by at least 90%, irrespective of the type of metered dose inhaler. This result suggests that spacer devices are useful in reducing local side effects in the oropharynx. In the stage assumed to represent the lung, the distribution ratio achieved with the Becotide inhaler with Volumatic was greater than that with the Aldecin with InspirEase. This study suggests that the best therapeutic effects can be expected from a combination of Becotide inhaler and Volumatic.

Hypocholesterolemic Action of Dietary Grifolin on Rats Fed with a High-cholesterol Diet.

The hypocholesterolemic action of grifolin was investigated in terms of its structure-activity relationship with rats fed on a high-cholesterol diet. The results show that the structure of farnesylorcinol was required for the hypocholesterolemic action, and that the effect of grifolin might be elicited, at least in part, through the augmented excretion of cholesterol into the feces.

Inhibitory effect of beta-carotene and astaxanthin on photosensitized oxidation of phospholipid bilayers.

Large unilamellar liposomes comprising of egg yolk phosphatidylcholine (PC) was exposed to photoirradiation in the presence of methylene blue (water-soluble photosensitizer) or 12-(1-pyrene)dodecanoic acid (P-12, lipid-soluble photosensitizer) to estimate the inhibitory effect of beta-carotene and astaxanthin on photosensitized oxidation of phospholipid bilayers. Without sensitizers, astaxanthin decreased much slower than beta-carotene and other hydrocarbon carotenoids (lycopene, alpha-carotene). Astaxanthin lasted longer than beta-carotene even in the presence of methylene blue or P-12. Decrease of astaxanthin was also much slower than that of beta-carotene when egg yolk PC was replaced by dimyristoyl PC. However, inhibitory effect of astaxanthin was lower than beta-carotene in the case of P-12 sensitized photooxidation. These results suggest that effectiveness of carotenoids as antioxidants on photosensitized oxidation (Type II) in phospholipid bilayers depends on the site of singlet oxygen to be generated, as well as their stability on photoirradiation.

Consumption of carotenoids in photosensitized oxidation of human plasma and plasma low-density lipoprotein.

Human plasma and plasma low-density lipoprotein (LDL) were exposed to photoirradiation in the presence of methylene blue (water-soluble photosensitizer) or 12-(1-pyrene)dodecanoic acid (P-12, lipid-soluble photosensitizer). In methylene-blue-sensitized photooxidation of human plasma and LDL, endogenous carotenoids and tocopherols were consumed with the accumulation of cholesteryl ester hydroperoxides (CE-OOH). Xanthophylls (zeaxanthin and lutein) decreased faster than lycopene and carotenes in the case of human plasma. In P-12-sensitized photooxidation of human plasma and LDL, the decrease rate of xanthophylls was slower than that of lycopene and carotenes. A lower level of beta-carotene exerted the effective inhibition of lipid peroxidation and retarded the oxidative loss of alpha-tocopherol, when the phosphatidylcholine liposomes containing these two lipid-soluble antioxidants were subjected to methylene blue- or P-12-sensitized photooxidation. These results suggest that antioxidant activity of carotenoids in photosensitized oxidation (Type II) of human plasma LDL depends on the site of singlet oxygen (1O2) to be generated and that carotenoids can protect tocopherols from the oxidative loss by 1O2 in the plasma.

Tracheobronchial mucociliary clearance and alveolar epithelial permeability measured by PET with 18FDG powder.

Tracheobronchial mucociliary clearance and alveolar permeability were measured with positron emission tomography using a water-soluble dry aerosol, sodium-N-acetyl-neuraminate tagged with 2-deoxy-2-[18F]fluoro-D-glucose (18FDG powder). Five normal volunteers inhaled 1.5-2.0 mCi of FDG powder by a single deep breath. The distribution of radioactivity, measured by scanning during a period of 120 min, showed that the 18FDG powder deposition progressed from the central airways to the peripheral alveolar areas. In the tracheobronchial system the radioactivity decreased to 24% of the initial deposition at 60 min. In the peripheral alveolar area, where absorption into blood or lymphatic flow crossing the epithelial layer represents a unique mechanism of clearance for water-soluble 18FDG powder, alveolar radioactivity decreased slowly to approximately 70% of the initial deposition at 60 min after inhalation. Positron emission tomography using 18FDG powder provides a regional evaluation of both mucociliary transport in the tracheobronchial system and epithelial permeability in the alveolar area.

Preparation of a fine powder of 2-deoxy-2-[18F]fluoro-D-glucose suitable for inhalation to diagnose lung diseases by means of PET.

Fine 2-deoxy-2-[18F]fluoro-D-glucose (18FDG) powder was obtained by adding diethyl ether into a methyl alcohol solution of 18FDG and other sugar as seed. When micronized particles of sodium N-acetyl-neuraminate (Neu5Ac-Na) were used as seed crystals, particles containing 18FDG were obtained and 80% of them were smaller than 10 microns in size. More than 60% of these crystals were 4-6 microns in size. In a preclinical study of forced inhalation in a dog, the 18FDG fine powder was mainly distributed in the trachea. The radioactivity in the trachea then increased once and a gradual decrease followed. The radioactivity was transferred into the blood and radioactivity incorporation into the heart was observed. After a normal volunteer inhaled 18FDG dry powder aerosol, the radioactivity was found in the respiratory tract and the peripheral area of the lung by means of PET. Absorption and in vivo dynamics of the 18FDG were also analysed.

Analysis of the effect of an anti-inflammatory steroid, dexamethasone, on neutrophil chemotaxis in the Boyden chamber with a modified 51Cr-labeling method.

For the determination of leukocyte chemotaxis in the Boyden chamber toward zymosan-activated serum, a modification of the 51Cr-labeling method originally introduced by Gallin et al. was made using a polycarbonate filter at the top of a double filter system. By application of this method, it was demonstrated that an anti-inflammatory steroid, dexamethasone at relatively low doses, directly inhibited chemotactic movement of leukocytes after the pretreatment of the cells with dexamethasone for several hours. This inhibitory effect was proved to be a common property of glucocorticoids. The leukocyte-inhibitory effect manifested within the cell body and the release of any extracellular factors mediating the inhibitory effect of the corticoid into the medium was not demonstrated. The present data suggests that the inhibitory effect of dexamethasone on leukocyte infiltration observed in vivo is attributed to the direct inhibitory effect of dexamethasone on the motility of leukocytes.

Mechanisms of anti-inflammatory action of dexamethasone: blockade by hydrocortisone mesylate and actinomycin D of the inhibitory effect of dexamethasone on leukocyte infiltration in inflammatory sites.

The present study was designed to clarify molecular mechanisms underlying inhibitory effect of dexamethasone on leukocyte infiltration in the inflammatory site. For the assay of leukocyte infiltration, two or four blebs were made s.c. on the back of rats by injecting with 2% carboxymethyl cellulose solution containing a chemoattractant, casein. Leukocyte accumulation in the bleb was inhibited considerably by local application of dexamethasone at a concentration of 0.6 X 10(-6) M. Hydrocortisone mesylate, which was reported in the study with hepatoma tissue culture cells to be a long-acting antagonist against glucocorticoid in binding to the corticoid receptor, blocked the above leukocyte inhibitory effect of dexamethasone when applied simultaneously with dexamethasone. The leukocyte infiltration was unaffected by the application of hydrocortisone mesylate alone. Treatment with androstenedione, which was reported to be inactive in the hepatoma tissue culture cells, did not interfere with the inhibitory effect of dexamethasone at all. Actinomycin D, when applied simultaneously with dexamethasone, significantly suppressed the leukocyte-inhibitory effect of dexamethasone. In contrast with those observations, the inhibitory effect of dexamethasone was not affected at all in cases that actinomycin D and hydrocortisone mesylate, respectively, were applied after the administration of dexamethasone. These results indicate essential roles of glucocorticoid receptor and gene expression for the manifestation of the inhibitory effect of dexamethasone on leukocyte infiltration in the inflammatory site.

Chemotactic factor production by rat polymorphonuclear leukocytes: stimulation with opsonized zymosan particles and inhibition by dexamethasone.

Rat polymorphonuclear leukocytes exposed to opsonised zymosan particles in vitro instantaneously and continuously release a chemotactic factor in the medium. The activity of this factor was mainly attributed to leukotriene B4, based on the data with high performance liquid chromatography. Preincubation of the cells with an antiinflammatory steroid, dexamethasone, at a dose of 0.25 micrograms/ml caused suppression in generation of the chemotactic factor from the leukocytes in a time-dependent manner.