Nedocromil sodium reduces cigarette smoke-induced bronchoconstrictor hyperresponsiveness to substance P in the guinea-pig.

Acute exposure to cigarette smoke provokes airway hyperresponsiveness to substance P and inactivates neutral endopeptidase (NEP). To determine whether nedocromil sodium can prevent cigarette smoke-induced hyperresponsiveness to substance P, we studied two groups of anaesthetized guinea-pigs. One group of guinea-pigs was pretreated with aerosolized 0.9% NaCl solution (90 breaths), the other group was pretreated with aerosolized nedocromil sodium (10(-4) M, 90 breaths). In each animal, pretreatment was followed by either exposure to the smoke of one cigarette or exposure to air. After acute exposure to cigarette smoke or to air, we measured the change in total pulmonary resistance (RL) induced by increasing concentrations of aerosolized substance P. In the absence of nedocromil sodium, the bronchoconstrictor responses to substance P were greater in cigarette smoke-exposed guinea-pigs than in air-exposed animals. Aerosolized nedocromil sodium had no effect on the response to substance P in air-exposed animals, but it reduced cigarette smoke-induced hyperresponsiveness to substance P. The preventive effect on cigarette smoke-induced hyperresponsiveness to substance P was observed at concentrations of aerosolized nedocromil sodium of 3 x 10(-5), 10(-4), and 3 x 10(-4) M. In vitro, cigarette smoke solution inhibited NEP activity from lung membrane preparations, but this inhibitory effect was not modified by nedocromil sodium (10(-4) M). We conclude that aerosolized nedocromil sodium reduces cigarette smoke-induced airway hyperresponsiveness to substance P in vivo. This action of nedocromil sodium is not due to a protective effect on cigarette smoke-induced inactivation of NEP in vitro.

Respiratory epithelial permeability in complete Freund's adjuvant rat lung granulomatosis.

To investigate Freund's adjuvant-induced changes in respiratory epithelial permeability, we determined the respiratory epithelium clearance of 99mTc-DTPA (RC-DTPA) in rats and the localization of horseradish peroxidase (HRP) across the lung epithelium. RC-DTPA was determined before and after intravenous injection of saline (control) or complete Freund's adjuvant (CFA). Although the respiratory epithelium was not disrupted, RC-DTPA was increased 15 min after injection of CFA and was still high after 3 d but had returned to normal after 7 d, after which it again increased and stayed high from Day 42 through Day 112. Cell number obtained by bronchoalveolar lavage increased from 15 min to 3 d after injection of CFA. Lung granulomatosis was observed 14 to 112 d after injection of CFA; changes in RC-DTPA correlated with changes in alveolar cells. HRP was present in bronchiolar intercellular spaces only in the CFA group. We conclude that (1) RC-DTPA increases in rat lung CFA granulomatosis 15 min to 3 and 42 to 112 d following injection, (2) the increase in RC-DTPA is not related to a rupture of epithelium but (3) is proportional to the cell influx into the lung, and (4) is possibly caused by the opening of tight junctions in small airways.

Tachykinins increase respiratory clearance of 99mTc-DTPA. Mechanisms of action.

We investigated the effects of tachykinins on the respiratory clearance of 99mTc-DTPA (RC-DTPA) in anesthetized and ventilated guinea pigs. We measured the change in RC-DTPA and in maximal pulmonary insufflation pressure (PIPmax) induced by substance P, neurokinin A, and capsaicin. Substance P, neurokinin A, and capsaicin increased both PIPmax and RC-DTPA in a concentration-dependent fashion. Substance-P- and capsaicin-induced increases in RC-DTPA were unaffected by pretreatment with atropine. Bilateral vagotomy attenuated substance-P-induced change in both RC-DTPA and PIPmax by approximately 70 and 50%, respectively. Capsaicin-induced change in RC-DTPA and PIPmax were slightly but not significantly reduced by bilateral vagotomy. The bronchodilator, salbutamol, dramatically reduced increase in RC-DTPA and in PIPmax induced by substance P, neurokinin A, and capsaicin, but it had no effect on increases in RC-DTPA and PIPmax generated by application of a positive end-expiratory pressure. We conclude that (1) tachykinins increase respiratory clearance to the solute, and (2) tachykinin-induced increase in RC-DTPA is not mediated by cholinergic neurotransmission but rather by the bronchoconstrictor effect of neuropeptides.

Influence of Pneumocystis carinii pneumonia on serum and tissue concentrations of pentamidine administered to rats by tracheal injections.

Pentamidine isethionate was administered by the tracheal route to control rats and immunodepressed rats with Pneumocystis carinii pneumonia (PCP). The serum concentration of pentamidine base 20 min after the administration was higher in the PCP rats (309 +/- 165 ng/ml) than in the control animals (71 +/- 36 mg/ml; p less than 0.001); 90 min after the injection the proportion of the pentamidine administered was lower in the right lung of the PCP rats (29 +/- 15%) than in the control rats (57 +/- 23%; p = 0.038); the proportion of pentamidine in the left kidney was higher in the PCP rats (14 +/- 4%) than in the control animals (4 +/- 2%; p less than 0.001). Respiratory clearance of 99mTc-DTPA, an index of the permeability of the respiratory epithelium, was higher in the PCP rats (1.84 +/- 0.42 %/min) than in the controls (0.44 +/- 0.11 %/min; p less than 0.001). We conclude that the more rapid diffusion of pentamidine from the alveolar lumen to the pulmonary circulation is explained by the increased alveolocapillary permeability as a result of pneumocystosis.

Effects of tobacco smoke on respiratory epithelial clearance of DTPA and on lung histology in rats.

The respiratory epithelial clearance of 99mTc-(DTPA) diethylenetriamine penta-acetate (RC-DTPA) was measured in rats before and after sham or cigarette smoke exposures. RC-DTPA was increased by cigarette smoke exposure (p less than 0.001); the amount and duration of the exposure had no significant effect. In smoker animals, the number of bronchoalveolar macrophages was decreased after one or five smoke exposures a day for one day (p less than 0.001) and the number of bronchoalveolar neutrophils was increased after five exposures a day for ten days (p less than 0.001). No abnormality was found in the lung parenchyma on light microscopy. We conclude that in rats RC-DTPA is increased by exposure to tobacco smoke, whatever the amount and duration of exposure; however, no gross parenchymal lung abnormality explains the increase in RC-DTPA.

Selective IgG subclass deficiencies and antibody responses to pneumococcal capsular polysaccharide antigen in adult community-acquired pneumonia.

We measured the serum concentrations of IgG subclasses in healthy subjects (n = 26) and in patients with community-acquired pneumonia (CAP) on admission (n = 38), at recovery (n = 21), and 9 months after admission (n = 19). Then, in 8 of the control subjects and 15 of the patients, we measured IgG subclasses and mean serum antibody concentrations of pneumococcal capsular polysaccharides before and 3 wk after immunization with a pneumococcal vaccine. Compared to the control subjects, the serum concentration of the IgG2 subclass was lower at admission in patients with CAP of bacterial or unknown cause (p less than 0.005). Concentrations of IgG subclasses in patients did not differ between admission and recovery, or between admission and 9 months later. After vaccination, in both control subjects and patients, there was an increase in the concentrations of IgG2 subclasses (p = 0.01) and antipneumococcal antibodies (p less than 10(-4)). We show that serum IgG2 concentration in patients with CAP of bacterial or unknown cause is lower than in healthy subjects and remains lower for several months. After immunization with a pneumococcal vaccine, the increase in serum concentrations of IgG subclasses and antipneumococcal antibodies in patients does not differ from those in control subjects.

Elastase activity of bronchoalveolar cells in advanced pulmonary sarcoidosis.

Elastase activities were investigated in bronchoalveolar lavage (BAL) cells from healthy controls (C, n = 8) and patients with untreated mediastinopulmonary sarcoidosis at different stages: I (n = 5), II (n = 7), IIIA (n = 3), IIIB (n = 4). Elastase activities (EA and ELA) were measured against a synthetic substrate, succinyl-trialanine-paranitroanilide (SLAPN) and radiolabelled ligamentum nuchae 3H insoluble elastin (specific radioactivity 3.7 MBq.mg-1). A significant increase of ELA of BAL cells was observed in patients (S III), in particular those exhibiting fibrotic roentgenographic patterns and a higher number of polymorphonuclear cells. On the contrary, EA of BAL cells showed no variation between different stages of sarcoidosis. ELA was never detected in the macrophage culture medium and appeared to be mainly associated with non-adherent macrophages, whereas EA appeared to be preferentially associated with adherent macrophages. The inhibitory profiles of ELA and EA were investigated using general proteinase inhibitors and also compounds interacting specifically with leucocyte elastase. ELA was mainly due to a serine protease, which has a sensitivity to inhibitors similar to leucocyte elastase and is increased in advanced stages of sarcoidosis. EA appears to be only slightly modified, if at all, in sarcoidosis and is probably related to the action of both a serine protease and a metalloenzyme. Thus, only ELA yields diagnostic information concerning stage III of sarcoidosis associated with a fibrotic or bullous roentgenographic pattern.

Comparison between theophylline analysis by nephelometric inhibition immunoassay and high performance liquid chromatography.

Rate nephelometric inhibition immunoassay (NIIA) was used to determine 94 serum theophylline concentrations, and these results were compared with those obtained using high performance liquid chromatography (HPLC) as a reference method. The measurements obtained by the nephelometric method were, on the average, 20% higher than those obtained by the chromatographic method (p less than 0.001). The coefficient of variation of the nephelometric method was 12.3%, compared with 3.9% for the chromatographic reference method. In the group of subjects having a serum concentration of caffeine greater than or equal to 0.5 microgram ml-1, the difference in serum theophylline concentration found between NIIA and HPLC correlated with serum caffeine concentration (r = 0.3755, df = 46, p less than 0.01). NIIA theophylline concentration was 8.8 +/- 3.2 micrograms ml-1 in serum from six additional patients receiving theophylline 9.1 +/- 3.4 micrograms ml-1 after adding 10 micrograms caffeine (NS), and 13.1 +/- 3.5 micrograms ml-1 after adding 10 micrograms 1,3-dimethyluric acid (p less than 0.001). We conclude that (a) the results obtained by the NIIA method are more variable and consistently higher than those obtained by the HPLC method and (b) 1,3-dimethyluric acid (a caffeine and theophylline metabolite) is responsible for this overevaluation.

Processing of lung lavage fluid causes variability in bronchoalveolar cell count.

To study the effects on cell counts of different ways of processing bronchoalveolar lavage (BAL) fluid, we performed 77 BAL in 19 healthy control subjects and in 58 patients with sarcoidosis. We investigated the role of readers, hemocytometers, mode of identification of macrophages, cell washing, and speed of cytocentrifugation. No significant effect of reader or hemocytometer was observed. The percentage of macrophages, determined as large cells stained with neutral red in a Malassez hemocytometer (63.9 +/- 24.6%, mean +/- SD) was lower than the percentage of macrophages determined by May Grunwald Giemsa staining (76.3 +/- 19.2%, p less than 10(-9)). Cell counts decreased 34% after 2 washings (p less than 0.001), and more lymphocytes were counted after cytocentrifugation at 90 g (33.2 +/- 25.3%) than at 23 g (27.7 +/- 22.1%). We conclude that bronchoalveolar cell counts vary with changes in processing lung lavage fluid and that this variability should be considered when using BAL cell counts.

[Respiratory permeability studies by clearance of an aerosol solute and bronchoalveolar lavage in interstitial pneumopathies]. / Perméabilité respiratoire déterminée par la clairance d'un soluté aérosolisé et le lavage bronchoalvéolaire dans les pneumopathies interstitielles.

Two methods of assessing the permeability of the respiratory epithelium have been compared: the ratio of serum albumin concentration to bronchoalveolar albumin (BA/S Alb) and the clearance of the aerosolized 99mTc-diethylene-triamine-penta-acetic acid (Tc-DTPA). The study population consisted of a control group of 21 normal subjects (group C1) in whom BA/S Alb was determined, a control group of 26 normal subjects (group C2) in whom the clearance of Tc-DTPA was measured and a group of 27 patients with interstitial lung disease including 20 with sarcoidosis (group PI) in whom both BA/S Alb and the clearance of Tc-DTPA were measured. BA/S Alb was higher in group PI (3.55 +/- 2.44 mg X g-1) than in group C1 (0.67 +/- 0.31 mg X g-1; p less than 0.001). The clearance of Tc-DTPA was higher in group PI (3.64 +/- 4.05% X min-1) than in group C2 (1.18 +/- 0.31% X min-1; p less than 0.01). BA/S Alb value exceeded the normal value of 1.33 mg X g-1 in 23 of the 27 patients, but the clearance of Tc-DTPA exceeded the normal value of 1.82% X min-1 in 15 of the 27 patients (X2 = 6.4; p less than 0.02). No correlation has been observed between BA/S Alb and the clearance of Tc-DTPA.(ABSTRACT TRUNCATED AT 250 WORDS)

Serum activity of angiotensin converting enzyme and pulmonary radiography as prognostic criteria in sarcoidosis.

To compare the prognostic value of the serum concentration of angiotensin converting enzyme (SACE) to that of pulmonary radiography, patients with mediastino-pulmonary sarcoidosis were followed over two years and then retrospectively placed in one of 2 groups, designated "cured" (CS) or "not cured" (NCS) on the basis of clinical and biologic criteria. The two groups were then compared with respect to the initial pulmonary radiologic image, and the initial SACE concentration. The initial SACE concentration (CS: 45.6 +/- 15.4 U . ml-1; NCS: 48.5 +/- 27.3 U . ml-1; means +/- SD) and the percentage of patients with elevated SACE (CS: 75%; NCS: 69%) did not differ significantly between the two groups; on the other hand, the percentage of patients with pulmonary nodules on their initial chest film was higher in the NCS group (50%) than in the CS group (21%; p less than 0.05). We conclude that pulmonary radiography is the more reliable prognostic criterion in mediastino-pulmonary sarcoidosis.

[Test of human basophil degranulation. The interpretation and analysis of variability factors in the absence of antigen]. / Test de dégranulation des basophiles humains. Interprétation et analyse des facteurs de variabilité en l'absence d'antigène.

The human basophil degranulation test (TDBH) is performed using a concentrated cell suspension enriched for polymorphonuclear basophils. This preparation is placed in wells on a glass slide with an allergen solution of decreasing concentration. After incubation the cells are fixed in the wells and stained with toluidine blue. Preparations containing fewer metachromatic basophils than in control preparation of the same cell suspension without antigen contain degranulated basophils and in this way a degranulation index can be calculated. By considering the variability in the number of basophils between duplicate control wells from the same cell suspension, it can be shown that the test can be considered to be positive only if the number of degranulated basophils equals or exceeds 50% of the total number of basophils. This variance can be attributed to the subjectivity of the reader, and to differences in the differential cell counts from well to well. On the other hand, the presence of atopic allergy had no effect on the basophil counts.

Elevation of the bronchoalveolar concentration of angiotensin I converting enzyme in sarcoidosis.

The concentration of angiotensin converting enzyme (ACE) and that of albumin (AIb) were assayed in the serum (SACE, SAlb) and in bronchoalveolar lavage fluid (LACE, LAlb). Three groups of patients were studied: 14 healthy volunteers (Group I), 45 patients with active sarcoidosis (Group II), and 7 patients with sarcoidosis in remission (Group III). The SACE in Group II (4,466 +/- 2,202 U/100 ml, mean +/- SD) was higher (p less than 0.001) than in Group I (2,470 +/- 547 U/100 ml) or in Group III (2,640 +/- 610 U/100 ml); LACE was higher in Group II (65.2 +/- 48.4 U/100 ml, p less than 0.001) than in Group I (21.1 +/- 14.7 U/100 ml), or in Group III (25.7 +/- 14.6 U/100 ml). The SAlb was found to be, respectively, 3,908 +/- 385 mg/100 ml, 3,982 +/- 965 mg/100 ml, and 3.613 +/- 222 mg/100 ml in Groups I, II, and III. The LAlb in Group II (8.2 +/- 6.2 mg/100 ml) was higher (p less than 0.01) than in Group I (2.5 +/- 1.4 mg/100 ml) or in Group III )4.1 +/- 1.0 mg/100 ml). The LACE in Group II increased with the number of alveolar lymphocytes, in nonsmokers (4 = + 0.56, df = 34, p less than 0.001) and in smokers (4 = + 0.88, df = 7, p less than 0.01). In the smokers in this group, LACE was higher with respect to the number of lymphocytes than in the nonsmokers. We conclude from this study (1) that the permeability of the alveolocapillary membrane to albumin and to ACE is increased in active pulmonary sarcoidosis, (2) that LACE increases during sarcoidosis and returns to normal when the disease is cured, and (3) that the concentration of ACE in alveolar fluid increases with tobacco use.

[Increased concentration of angiotensin I converting enzyme in the alveolar fluid of patients with sarcoidosis]. / Augmentation de la concentration de l'enzyme de conversion de l'angiotensine I dans le fluide alvéolaire de sujets sarcoidiens.

Angiotensin converting enzyme (ACE) was assayed both in serum (SACE) and in bronchoalveolar fluid lavage (LACE) in 14 healthy controls and in 45 patients with sarcoidosis with mediastinal and pulmonary involvement. Concentration of SACE was 4466 +/- 2202 U x 100 ml-1 (mean +/- SD) in sarcoidosis and 2470 +/- 547 U x 100 ml-1 (chi +/- SD) in sarcoidosis and 2470 +/- 547 U . 100 ml-1 in controls. Concentrations of LACE were 65.2 +/- 48.4 U . 100 ml-1 and 21.1 +/- 14.7 U . 100 ml-1 respectively in sarcoidosis and in controls. These results are in favor of an intraalveolar secretion of ACE in sarcoidosis. LACE could be a better criterium than SACE for the evaluation of the pulmonary activity of sarcoidosis.

COHb formation and acute carbon monoxide intoxication in adult male rats and guinea-pigs.

Respiratory exchange recordings and percentage survivals to lethal concentrations show that rats are more sensitive than guinea-pigs to an acute carbon monoxide intoxication. To avoid circadian respiratory an comportmental differences between rats and guinea-pigs experiments were performed on urethane anesthetized and artificially ventilated animals. This procedure demonstrates that, for an inhaled concentration of 0.05% CO, the rates of formation of COHb do not statistically differ in rats and in guinea-pigs, but that a higher (0.01 less than p less than 0.05) COHb saturation is reached in rats (35%) than in guinea-pigs (25%). For a 2.84% CO inhalation, no statistically significant difference is observed in the rate of COHb formation, but cardiac arrest is sooner (p less than 0.001) observed in rats than in guinea-pigs.

Immunoglobulin E in human aqueous humor and corresponding serum. A physiopathological and clinical study.

The analysis of IgE in aqueous humor yielded an average concentration of 3.4 +/- 0.97 U/ml for 22 cataract patients and 5.5 +/- 3.42 U/ml for five uveitis patients. The IgE level in aqueous humor (IgEa.h.) of the cases examined is most probably, beside hematoocular diffusion of serum IgEs, the result of intra-ocular IgE production. In comparison with (mostly normal) IgEs levels, the IgEa.h. concentration appears relatively elevated, not only with uveitis patients, but also with cataract patients, above all when lenticular opacity is accompanied by other ophthalmic diseases (glaucoma, high myopia, diabetes). This "increase" of IgEa.h. concentration in very probably due to the radioimmunosorbent (RIST) technique employed, the most sensitive method available at the time of the present study. Thus, the calculated IgEa.h. value in the cataractous eyes should be regarded simply as approximate to the normal IgEa.h. concentration. These values are of clinical significance however, since a reference IgEa.h. mean-value is indispensable to the interpretation of pathologically high IgEa.h. levels and ethics do not permit of IgEa.h. determination in healthy eyes. The mean IgEa.h. levels of the delayed-type uveitis and cataract patients examined reveal no significant differences. IgEa.h. determination could make a contribution to the etiological clarification of, for example, immediate-type uveitis cases and intra-ocular parasitosis and serve as an appropriate model to study intra-ocular immunomechanisms.