Purification and level of expression in bronchoalveolar lavage of a human polychlorinated biphenyl (PCB)-binding protein: evidence for a structural and functional kinship to the multihormonally regulated protein uteroglobin.

A human lung polychlorinated biphenyl (PCB)-binding protein was purified by sequential chromatography of lavage fluid incubated with the tritium-labeled, high-affinity ligand, 4,4'-bis(methylsulfonyl)-2,2',5,5'-tetrachlorobiphenyl. From sodium dodecyl sulfate polyacrylamide gel electrophoresis gradient gels, it was evident that a single band with an approximate molecular weight of 13 kD was present in the eluate from the final chromatographic step. Antibodies raised against the human lung PCB-binding protein detected a single band of corresponding size in lavage fluid in immunoblotting experiments. Furthermore, the antibodies detected significantly higher levels of the lung PCB-binding protein in lavage fluid from nonsmokers as compared to smokers. The purified protein was sequenced, and an alignment of the obtained aminoterminal amino acid residues of the human lung PCB-binding protein to uteroglobin and to a rat lung PCB-binding protein revealed an overall positional identity of approximately 45%. The amino acids suggested to participate in ligand binding of uteroglobin were extensively conserved in the PCB-binding proteins. Thus, we conclude that we have purified and raised antibodies against a human lung PCB-binding protein and that it has a structural as well as a functional kinship to the steroid-binding and multihormonally regulated rabbit protein uteroglobin.

Glucocorticoid receptor mRNA expression in pulmonary alveolar macrophages in sarcoidosis.

The presence of the glucocorticoid receptor was demonstrated by immunocytochemistry in pulmonary alveolar macrophages obtained by bronchoalveolar lavage. Also, GR mRNA content was determined by solution hybridization in PAM from 12 healthy volunteers and in 6 patients with sarcoidosis. No significant differences with regard to GR mRNA expression was detected between the two groups examined. For comparison, lung tissue from three patients undergoing thoracic surgery was examined and found to contain GR mRNA levels in the same range. As an indication of GR function, we also determined the mRNA levels of a glucocorticoid-regulated gene, metallothionein IIA, during basal conditions and after in vitro incubation of PAM with dexamethasone. Neither the control sample nor the dexamethasone-stimulated MTII mRNA values in PAMs differed significantly between the two groups. Solution hybridization is a rapid, sensitive and convenient assay which enables accurate and specific quantitation of GR mRNA in PAM. The GR mRNA content and basal as well as dexamethasone-induced MTII mRNA levels in PAM from patients with sarcoidosis is not significantly different from those in healthy volunteers.

The use of a vasodilator, felodipine, as an adjuvant to long-term oxygen treatment in COLD patients.

Eight patients with chronic obstructive lung disease (COLD) and pulmonary hypertension were given an infusion of a calcium antagonist, felodipine, during ongoing, long-term oxygen treatment (LTOT). The effects on central haemodynamics and ventilation-perfusion matching were studied. At rest pulmonary and systemic vascular resistances (PVR and SVR) were reduced by 18% (NS) and 26% (p less than 0.05), respectively. Cardiac output increased by 23%. There was a tendency to increased perfusion of low alveolar ventilation-perfusion ratio (VA/Q) areas (VA/Q less than 0.1) and to increased shunt compared to pretreatment values. Arterial oxygen tension (PaO2) fell by 0.7 kPa (p less than 0.001) but total oxygen transport increased by 23% (p less than 0.001). After treatment with oral felodipine (7.5-15 mg.day-1) for a mean time of 14 wks, PVR and SVR were reduced by 16% (p less than 0.05) and 7% (NS), respectively, as compared to pretreatment values at rest. Cardiac output rose by 13%. The VA/Q ratios and the PaO2 returned towards pretreatment values. The total oxygen transport increased by 11% (p less than 0.05) at rest and increased by 19% (p less than 0.05) during exercise as compared to the pretreatment value. The positive effect on central haemodynamics indicates that felodipine may be a valuable adjunct to ongoing LTOT.

Stimulation of bronchoalveolar lavage (BAL) and blood lymphocytes by Kveim antigen, tuberculin and concanavalin A in sarcoidosis.

BAL and blood mononuclear cells and their reactivity to Kveim antigen, tuberculin and concanavalin A (Con A) were studied in nine patients with different clinical stages of sarcoidosis. After separation by plastic adherence, non-adherent cells (mainly lymphocytes) were admixed with 10% autologous adherent cells (monocytes/macrophages). After 3 and 6 days' culture with Kveim antigen (1, 10, 100 micrograms/ml), PPD tuberculin (2.5 micrograms/ml) and Con A (10, 20, 40 micrograms/ml) stimulation was measured as incorporation of 14C-thymidine into DNA. Except for occasional reactions the study did not show any unitary significant increase in lymphocyte response to the different concentrations of Kveim antigen in either BAL or blood. For Con A there was a weaker response by BAL-mononuclear cells with no difference between 3 and 6 days, compared with blood where there was an early peak. The lymphocyte reaction to PPD was weak with no difference between blood and BAL.

Cardiac function and central haemodynamics in severe chronic obstructive lung disease. Acute and long-term effects of felodipine.

Eleven patients, with advanced chronic obstructive lung disease (COLD), received an infusion of the calcium antagonist felodipine at a rate of 0.9 mg/h. Pulmonary and systemic vascular resistances (PVR and SVR) at rest were reduced by 18% (p less than 0.05) and 33% (p less than 0.001), respectively. Cardiac output increased by 33%. The right ventricular and left ventricular ejection fractions (RVEF and LVEF), measured by equilibrium gated radionuclide ventriculography, increased by 32% (p less than 0.01) and 25% (p less than 0.01), respectively. During exercise both PVR and SVR fell by a mean of 30% (p less than 0.01). RVEF and LVEF both increased by about 14% (p less than 0.05 and p less than 0.01). After three months of oral felodipine treatment, a dose-related decrease in PVR was noted at rest (r = -0.83) compared with pretreatment values. There was an increase in RVEF which correlated to a reduction in PVR (r = -0.76). Three patients discontinued the trial due to side effects. It is concluded that the reduction of PVR induced by felodipine is accompanied by an improvement in right heart function as measured by ejection fraction measurements.

Mutagenicity studies by co-cultivation of bronchoalveolar cells and blood lymphocytes with V79 Chinese hamster cells.

Human bronchoalveolar cells, consisting of approximately 85% pulmonary alveolar macrophages (PAMs), and peripheral blood lymphocytes isolated from healthy volunteers were investigated for their ability to metabolize 7,8-diol of benzo[a]pyrene (B(a)P). The mutagenicity of reactive metabolites was analyzed by employing a co-cultivation system using V79 Chinese hamster cells for the detection of mutations. The metabolic activity of the human cells was compared to PAMs isolated from rabbits. The number of PAMs obtained by bronchoalveolar lavage of smokers was found to be significantly elevated compared with nonsmokers. However, the mean number of induced mutations of the 7,8-diol mediated by PAMs during co-cultivation did not differ significantly between smokers and nonsmokers. The aryl hydrocarbon hydroxylase (AHH) activity of human lymphocytes has been studied by others, but to the best of our knowledge this is the first demonstration that human blood lymphocytes could be successfully used in a co-cultivation assay for the characterization of xenobiotic metabolism in terms of mutations, as illustrated by the linear increase of induced mutations in the V79 cells. Rabbit PAMs were less efficient in mediating mutations as compared to both smokers' and nonsmokers' PAMs or lymphocytes. This can probably be explained by less efficient bioactivation of 7,8-diol in rabbit PAMs, which is supported by the fact that the rabbit PAMs metabolized B(a)P in a different way as compared to human PAMs as revealed by HPLC analysis of ethyl acetate extractable metabolites of 3H-B(a)P. No qualitative or quantitative difference in the patterns of B(a)P metabolism by PAMs isolated from smokers and nonsmokers could be established. In conclusion, human PAMs were found to be more efficient in terms of cell-mediated mutagenicity than human lymphocytes, which are more efficient than rabbit PAMs. The present results differ from previous reports concerning the xenobiotic metabolizing capacity of these cells assessed by other methods. This illustrates the usefulness of the co-cultivation assay, because it measures not only the bioactivating capacity of isolated mammalian cells, but also their detoxifying capacity, the transfer of mutagens to other cells and the ability of their metabolites to cause mutations.

Theophylline in maintenance treatment of chronic asthma: concentration-dependent additional effect to beta 2-agonist therapy.

The effect of long-term treatment with theophylline was studied in 20 chronic asthmatic patients receiving oral and inhaled beta 2-agonists. In a double-blind, randomized cross-over fashion during three consecutive 6-week periods, the patients received theophylline in individually adjusted dosages or placebo to obtain plasma concentrations of 25-45 mumol/l, 50-85 mumol/l and 0, respectively. PEF, beta 2-aerosol consumption, symptom scores and side-effects were recorded daily. The addition of theophylline caused a significant further bronchodilating effect. There was, however, a large interindividual variation in response to additional theophylline and only half of the subjects were responders. Those responding had increased therapeutic efficacy from a steady-state concentration in the range of 50-85 mumol/l compared to the lower concentration. The responders had, on average, more pronounced air-flow obstruction. The incidence of adverse reactions increased with increasing concentrations of theophylline.

Characterization of a binding protein for the PCB metabolite 4,4'-bis(methylsulfonyl)-2,2',5,5'-tetrachlorobiphenyl present in bronchoalveolar lavage from healthy smokers and non-smokers.

Bronchoalveolar lavage (BAL) was performed in a group of healthy subjects (10 smokers and 10 non-smokers) and the recovered fluid was shown to contain specific binding sites for a metabolite of a polychlorinated biphenyl (PCB), 4,4'-bis ([3H]methylsulfonyl)-2,2',5,5'-tetrachlorobiphenyl [(3H-MeSO2)2TCB]. The sites seem to reside within a protein-like component and the apparent dissociation constant (Kd) for the binding was approximately 2 X 10(-7) M regardless of the smoking status of the subject. However, the maximum number of binding sites (Bmax) was significantly lower for the smokers (p less than 0.001). Competition studies indicated that some PCB methyl sulfones had similar affinities for the specific binding sites as (MeSO2)2TCB. Physicochemical characterization of the human (3H-MeSO2)2TCB-binding protein indicated a Stokes radius of 22 A and a sedimentation coefficient of 1.9 S, and on the basis of these parameters an apparent molecular weight of 17,700 was calculated. The binding protein had an apparent pI of 4.9. It is suggested that the specific binding protein for certain PCB methyl sulfones in bronchoalveolar lavage fluid from healthy subjects is responsible for the previously observed tendency of PCB metabolites to accumulate in human lung tissue.

Long-term treatment with a new calcium antagonist, felodipine, in chronic obstructive lung disease.

Nine patients with advanced chronic obstructive lung disease (COLD) were treated with a calcium antagonist, felodipine, for 3-5 months and their central haemodynamics and pulmonary gas exchange were then studied. The systemic vascular resistance was reduced by 19% (p less than 0.05) at rest and by 30% during ergometer bicycle exercise (p = 0.05) compared to pretreatment data, whereas pulmonary vascular resistance showed a borderline reduction of 10% at rest and 30% during exercise (p = 0.12 and p = 0.10, respectively). Stroke volume increased by 13% (p less than 0.05) at rest and to the same extent at exercise. A moderate deterioration of the ventilation-perfusion relationship was seen by the multiple inert gas elimination technique, and the arterial oxygen tension was further reduced by 3.5 mmHg (p less than 0.05) at rest and by 3.0 mmHg on exercise, but the working capacity on an ergometer bicycle increased from 60 to 70 W (p less than 0.01). No acceptable predictor of patients who would respond to the drug could be found.

Nasal hypersensitivity in wood furniture workers.

Occupational nasal allergies were studied in six wood furniture factories. The concentration of moulds, wood dust and endotoxins was registered and occasionally high values were found. Paecilomyces spec. was the most common mould. A special questionnaire showed that 16% (42/268) of the wood workers with a mean exposure time of 12 years had a history compatible with hypersensitivity in the upper airways associated to their work. Rhinomanometric investigations showed mucosal congestion in the workers with nasal discomfort and nasal clearance was pathologically slow in 54%. Histological studies revealed a high incidence of unciliated and metaplastic nasal epithelium. Skin prick tests and provocation tests with standard allergens and allergens prepared from the moulds and the wood dust in the environment showed that the wood furniture workers had an incidence of allergy to moulds in 3% and to wood dust in 2%. Most of the workers sensitive to moulds and/or woods were also skin prick sensitive to other allergens. No statistical difference concerning the presence of precipitating antibodies against mould and wood antigens could be registered between workers with discomfort and workers without symptoms.

The effect of a new calcium antagonist, felodipine, on pulmonary hypertension and gas exchange in chronic obstructive lung disease.

In 10 patients with advanced chronic lung disease and pulmonary hypertension central haemodynamics and pulmonary gas exchange were studied at rest and during exercise, before and during continuous infusion of the calcium antagonist felodipine, which lowers vascular resistance by selective action on vascular smooth muscle. Pulmonary and systemic vascular resistance were reduced by 20% and 35% respectively. Cardiac output was increased by 28% and pulmonary vascular pressures remained essentially unaltered. Gas exchange measurements revealed an increased shunt, an addition of lung regions with high ventilation/perfusion ratios and a further reduction of arterial oxygenation from 8.5 to 7.9 kPa (64 to 59 mmHg). During exercise, infusion of felodipine resulted in a higher cardiac output and a smaller increase in pulmonary vascular pressure than before felodipine. Arterial oxygenation fell to the same extent as during exercise before felodipine. It is concluded that felodipine lowers pulmonary vascular resistance but at the expense of a worsening of the ventilation-perfusion ratio.

Nasal hypersensitivity in wood furniture workers. An allergological and immunological investigation with special reference to mould and wood.

Occupational nasal allergies caused by moulds and wood dust were extensively studied in six wood furniture factories. The concentration of moulds, wood dust and endotoxins was registered and occasionally high values were found. Paecilomyces spec. was the most common mould. Answers to a special questionnaire showed that 16% (42/268) of the wood workers with a mean exposure time of 12 years had a history compatible with hypersensitivity in the upper airways associated with their work. Calculations based on skin prick tests and provocation tests with relevant allergens showed that the wood furniture workers had an incidence of allergy to moulds in 3% and to wood dust in 2%. Most of the workers (5/7) sensitive to moulds and/or woods were also skin prick sensitive to other allergens. No statistical difference concerning the presence of precipitating antibodies against mould and wood antigens could be registered between workers with discomfort and workers without symptoms.

Oral administration of terbutaline in asthmatic patients.

The absorption and excretion profiles of orally administered terbutaline were investigated in 10 asthmatic patients who received a solution containing 5 mg of [3H]terbutaline sulphate. Bronchodilating effect and side-effects were followed. The mean serum concentration curve reached its maximum of 3.0 +/- 0.3 (SEM) ng/mL at 60-90 min. Renal clearance data indicated that terbutaline was excreted only via glomerular filtration. Approximately 30% of the dose was excreted in the urine in 12 h and 40% in 72 h. Three compounds appeared in the urine: unchanged terbutaline, a sulphate conjugate, and a glucuronide, the main metabolite being the sulphate conjugate. The maximum mean increase in volume of air expelled in the first second of forced expiration (FEV1) was 35%, attained 90-120 min after intake of terbutaline. There was a correlation (r = 0.74) between the serum level and the bronchodilator effect. No significant effects on heart rate or blood pressure were found. Tremor, as objectively measured with the aid of an accelerometer, was experienced by the patients when it increased to twice its basal level. The observed tremor did not appear to parallel either the serum level or the bronchodilating effect. A comparative study in 3 of the patients showed that [3H]terbutaline formulated as a tablet had similar bioavailability.

Simultaneous treatment with terbutaline and theophylline.

The bronchodilating effect and side-effects of theophylline and a beta 2-adrenoceptor stimulating drug (terbutaline) alone and in combination were studied in 10 adult asthmatic patients. Initially, each individual's pharmacokinetic parameters for theophylline were determined. On 3 separate days, theophylline was infused to defined steady-state concentrations (0, 7.5, and 15 micrograms/mL, respectively) followed by the administration, at intervals of 1 h, of incremental intravenous doses of terbutaline. Theophylline caused a plasma concentration-dependent increase in the volume of air expelled in the first second of forced expiration (FEV1). Theophylline itself had no significant effect on objectively recorded skeletal muscle tremor or heart rate but enhanced the terbutaline-induced increase in both. At the highest steady-state concentration (15 micrograms/mL), 2 of the patients experienced nausea. Terbutaline caused a plasma concentration-dependent increase in FEV1, heart rate and tremor. The plasma levels of terbutaline at the given doses did not differ significantly between individuals or within individuals at different theophylline levels. The combination of theophylline and terbutaline resulted in an additive effect on bronchodilatation. At comparable levels of bronchodilatation, the combination of theophylline and terbutaline caused a lesser degree of side-effects than either drug alone.

Separate and combined use of terbutaline and theophylline in asthmatics. Effects related to plasma levels.

The bronchodilating properties and side effects of theophylline and a beta 2-adrenoceptor stimulating drug (terbutaline) alone and in combination were studied in 10 adult asthmatic patients. Initially, each individual's pharmacokinetic parameters for theophylline were determined. On 3 separate days theophylline was infused to defined steady-state concentrations (0, 7.5, and 15 microgram/ml, respectively) followed by the administration, at 1-h intervals, of incremental i.v. doses of terbutaline. It was shown that: 1. Theophylline caused a concentration-dependent increase in FEV1. Theophylline itself had no significant effect on objectively recorded skeletal muscle tremor or heart rate but enhanced the terbutaline-induced increase in tremor and heart rate. At the highest concentration (15 microgram/ml) two of the patients experienced nausea. 2. Terbutaline caused a concentration-dependent increase in FEV1, heart rate, and tremor. The plasma levels of terbutaline at the given doses did not differ significantly between individuals or within individuals at the different theophylline levels. 3. The combination of theophylline and terbutaline resulted in a mere additive effect on the bronchodilatation. At comparable bronchodilatation the combination of theophylline and terbutaline caused a lesser degree of side effects than each of the drugs alone.

Muscular hyperplasia of the lung: a clinical, radiographic, and histopathologic study.

The clinical, radiographic, and histopathologic characteristics of pulmonary muscular hyperplasia were analyzed in 13 patients who were seen over a 13 year period. The average follow-up period was 7.2 years (range, 1-13). Eight patients initially demonstrated interstitial disease radiographically and the radiographic diagnosis was "interstitial fibrosis." The radiographs of the other five patients exhibited only localized infiltrates initially--produced by bronchiectasis in three, inflammatory mass in one, and bronchial carcinoma in one. Histologic features were consistent with desquamative interstitial pneumonia in eight, usual interstitial pneumonia in one, and unspecified interstitial pneumonia in four. When tuberous sclerosis is absent, pulmonary muscular hyperplasia is probably an end stage of interstitial pneumonia.

In vitro culture of peripheral blood and bone marrow cells in patients with sarcoidosis with and without special preparations of Kveim antigen.

Colony-forming cells (CFC) and colony-forming ability in peripheral blood and bone marrow were studied in 22 patients with different stages of sarcoidosis, with or without the addition of different Kveim preparations. The study showed no statistical differences between the colony growth in healthy persons compared to patients with sarcoidosis in stages I and II. The counts in these groups were, however, lower than in patients with sarcoidosis in stage III (P < 0.01). The lowest counts of CFC were found in the highly active cases suffering from fever, joint pains and erythema nodosum, and these counts were also lower than generally found in normal cases. After stimulation with mononuclear cells from healthy controls the number of colonies in the acute stages, however, reached normal values. This suggests either a low autostimulation or a neutralization of an inhibiting factor in the blood. It is suggested that if an inhibiting factor is present it is principally localized to the sarcoid granuloma. A pronounced inhibition of colony growth was seen when the Kveim membrane fraction prepared from sarcoid lymph nodes and spleen, was added to the cell cultures.