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Late phase increase of thrombin-like proteinase, protein, leukocytes in bronchoalveolar lavage (BAL) fluid and chemiluminescence of BAL after ovalbumin aerosol inhalation by actively immunized rats.

A thrombin-like proteinase (THROLP) was detected colorimetrically as the main proteolytic activity (PROA) in bronchoalveolar lavage fluid (BALF) 4 and 24 h after ovalbumin aerosol (OVA) challenge of actively immunized rats. Other proinflammatory parameters increased also significantly in BALF, like chemiluminescence of leukocytes (1 h), protein and cell number (24 h after challenge). In parallel, increased bronchoconstrictions against 5-HT aerosols are detected 24 h post OVA challenge. THROLP is an indicator for plasma leakage, activation of the clotting reaction and the protracted inflammation in the airways, which induced bronchial hyperreactivity.

Administration, Inhalation

Bronchoalveolar lavage in the normal volunteer subject. 2. Safety and results of repeated BAL, and use in the assessment of intrasubject variability.

To investigate the safety of repeated bronchoalveolar lavage (BAL) and the variability of commonly measured parameters from BAL to BAL in the same subject, we performed a total of 59 BALs in 16 normal volunteer subjects. The BAL was performed with 120 ml (three aliquots of 40 ml) of room temperature, normal saline in a lingular subsegment. Four subjects had five BAL, three had four BAL, and nine had three BAL performed at minimal intervals of six weeks. The BAL analysis included percentage of lavageate returned, cell number, and percentage of alveolar macrophages, lymphocytes, neutrophils and eosinophils. Relatively similar percentages of lavageate were returned on each lavage. There was considerable variability in the cell numbers obtained both within and between subjects, although some subjects had consistently high, low, or normal cell numbers returned from each lavage. Cell differential was the most consistent parameter on repeated BAL analyses, but isolated "abnormal" elevations in the percentage of one or another cell type were occasionally noted. These were unrelated to either the number or relative sequence of the BALs. Pulmonary function tests performed both before and after the repeated BAL showed no significant change and participants noted no subjective deterioration in pulmonary function. This study supports the safety of repeated BALs in the normal subject, but the variability in cell numbers obtained and isolated, "abnormal" elevations of inflammatory cells occasionally noted in this normal population indicate that BAL parameters in patients need to be interpreted with extreme caution.

Adult

Elevated BAL fluid histamine levels and parenchymal pulmonary disease in rheumatoid arthritis.

To determine the amount of histamine in BAL fluid in subjects with RA and to ascertain if elevated histamine levels were associated with parameters of active pulmonary disease, we measured BAL fluid histamine levels in 31 subjects with RA and 36 normal subjects. The subjects with RA had a significantly greater mean BAL histamine level than the normal subjects, (313 +/- 154 pg/ml vs 18 +/- 8 pg/ml; p less than 0.05). When the subjects with RA were divided into three groups based on chest radiograms (1 = normal; 2 = pleural disease only; 3 = interstitial or nodular disease), we found that subjects in group 3 had significantly lower values for TLC and D. Subjects in group 3 also had higher percentages of BAL neutrophils and eosinophils and higher BAL histamine levels (group 1, 115 +/- 52 pg/ml; group 2, 30 +/- 30 pg/ml; and group 3, 1,182 +/- 709 pg of histamine per milliliter). Moreover, BAL histamine levels were negatively correlated with TLC (r = -0.46; p = 0.01) and FVC (r = -0.45; p = 0.01) and positively correlated with BAL neutrophils (r = 0.6; p = 0.0003) and BAL eosinophils (r = 0.89; p = 0.0001). These data suggest that the BAL histamine level may be a useful marker to determine the activity of pulmonary disease in RA.

Arthritis, Rheumatoid

[Influence of various factors on glucocorticoid receptors of bronchoalveolar lavage (BAL) cells from rabbits].

It is supposed that the effects of glucocorticoid on target cells are mediated by glucocorticoid receptors. Recent investigations have demonstrated that changes in the composition and function of immune cells in bronchoalveolar lavage (BAL) cells of patients with immunological lung diseases are associated with the intensity of alveolar septal inflammation, or indicate steroid responsiveness. In order to clarify the physiologic changes of glucocorticoid receptor (GR) we measured the GR content of BAL cells obtained from rabbits with various conditions of the host (aging, immunological activation, malnutrition and pretreatment with prednisolone). The specific binding to GR was carried out by incubating BAL cells with 1, 2, 4, 10, 20, 40 nM 3H-prednisolone in the presence or absence of 1.0 x 10(-6) M unlabeled prednisolone. BAL cells from rabbits of various ages exhibited age-dependent reduction of GR content. There were striking differences of GR content in BAL cells from normal and BCG-vaccinated rabbits. The former yielded 3124 +/- 365 binding sites per cell, whereas activated BAL cells showed considerably higher values (6807 +/- 766). Chronological changes in GR content in BAL cells were also observed in normal rabbits pretreated with intravenous injection of prednisolone (2 mg/kg). The GR content was lowest at 3 hours after administration, reaching a peak at two days and then returned to control level. As for nutritional modulation, GR content tended to increase in the initial phase of fasting (days 4 to 8), and then decreased after day 12. In contrast, there were no significant changes in dissociation constants among BAL cells from animals in normal, immunologically activated and malnourished conditions.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging

Interference by 2, 3-dimercapto-1-propanol (BAL) in angiotensin I radioimmunoassay.

The angiotensinase inhibitor 2,3-dimercaptopropanol (BAL) interferes with peptide-antibody binding when certain sensitive antisera are used in angiotensin I radioimmunoassay systems. Three of nine antisera tested showed sufficient interference to produce serious errors in data obtained using these antisera together with BAL. For PRA determinations in human plasma, at both pH 5.7 and pH 7.3, relationships between different PRA'S are altered, and results of renin stimulation tests are changed in unpredictable ways. Determination of renin concentration in rat plasma does not require use of BAL as inhibitor, and it is best avoided. For human plasma renin determinations, use of BAL-sensitive antisera should be avoided, since there is no satisfactory way to correct data for the resulting error. BAL itself, rather than its oxidation products, is probably the interfering substance. The interference appears to be due to an interaction between BAL and the BAL-sensitive antiserum. It is not related to the known actions of BAL as chelating or reducing agent.

Angiotensin II

Levels and specificity of antibody in bronchoalveolar lavage (BAL) and serum in an animal model of trimellitic anhydride-induced lung injury.

A study was undertaken to characterize the antibody response in rats exposed to trimellitic anhydride (TMA) by inhalation. Total antibody levels directed to trimellitic rat serum albumin (TM-RSA) from TMA-exposed rats were assayed by an ammonium sulfate technique. Total antibody levels in bronchoalveolar lavage (BAL) and the matched serum were compared by correction for the albumin content of each. An ELISA was developed to detect IgG, IgA, and IgM directed toward TM-RSA in BAL and serum and to compare class-specific antibody levels in BAL and serum by normalizing for albumin content. The specificity of the rat IgG response was determined by ELISA inhibition with TM-RSA and TM-human serum albumin (TM-HSA) and compared with reciprocal inhibition studies with serum from TMA-exposed workers. The levels of total antibody in BAL were three to 15 times greater than the levels found in the matched serum pair. IgG, IgA, and IgM antibodies were detected in the BAL and the serum of TMA-exposed rats but not in control rats. In each of the four rats tested, all antibody classes were present in equal or greater amounts in the BAL than in the serum. Complete inhibition of the rat IgG binding in ELISA was observed when TM-RSA or TM-HSA were added as inhibitors. Human IgG was inhibited in ELISA only by TM-HSA. In an animal model of human lung disease, the levels of total antibody as well as class-specific antibodies directed against TM-RSA were greater in BAL than in serum.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

The site of inhibition of the chloroplast electron-transport system by 2,3-dithiopropan-1-ol (BAL).

BAL (2,3-dithiopropan-1-ol) treatment of chloroplasts has previously been reported to induce a block in electron transport from water to NADP+ at a site preceding plastocyanin [Belkin et al. (1980) Biochim. Biophys. Acta 766, 563-569]. In the present work the block was further characterized. The following properties of BAL treatment are described. Inhibition of electron transport from water to lipophilic acceptors but not to silicomolybdate. Inhibition of the slow, sigmoidal phase of chlorophyll a fluorescence induction. Inability of N,N,N',N',-tetramethyl-p-phenylenediamine to bypass the inhibition of NADP+ photoreduction with water as the electron donor. Inhibition of electron transport from externally added quinols to NADP+. Inhibition of cytochrome f reduction by photosystem II, but not its oxidation by photosystem I. Inhibition of cytochrome b6 turnover and cytochrome f rereduction after single-turnover flash illumination under cyclic electron-flow conditions. The BAL-induced block is therefore located between the secondary quinone acceptor (QB) and the cytochrome b6f complex. It was further found that (a) the isolated cytochrome complex is not inhibited after BAL treatment; (b) BAL-reacted plastoquinone-1 inhibits electron transport in chloroplasts; (c) BAL does not inhibit electron transport in chromatophores of Rhodospirilum rubrum or Rhodopseudomonas capsulata. It is suggested that the inhibition of electron transport in chloroplasts results from specific reaction of BAL with the endogenous plastoquinone.

Bacterial Chromatophores

Arsenic distribution in rabbits after Lewisite administration and treatment with British anti-Lewisite (BAL).

The standard treatment of Lewisite (dichloro(2-chlorovinyl)arsine) poisoning is by chelation with BAL (British anti-Lewisite, dimercaptopropanol). The present study investigated the effect of BAL treatment on the distribution of arsenic after Lewisite administration. Lewisite was administered subcutaneously at the LD10 and LD40 of the compound. Without BAL treatment arsenic was eliminated with a half-life in blood of between 55 and 75 hr and a blood clearance of 120 ml/hr/kg. Arsenic had a large volume of distribution of several liters per kilogram, indicating extensive distribution in tissues. The highest tissue concentrations, more than seven times blood concentrations, were found in the liver, lung, and kidneys. These organs maintained an approximately constant concentration ratio with blood during the sampling period. Concentrations in tissues with a blood-to-tissue barrier, such as the brain and the spinal cord, rose between 4 and 96 hr while blood concentrations declined more than fourfold over the same time period. BAL treatment by four equal, maximally tolerated doses over 12 hr substantially reduced arsenic concentrations in blood and tissues. For example, at 24 hr the concentrations in brain and liver (target organs for arsenic toxicity) were reduced by 65 to 89% over the range of Lewisite doses administered. The total exposure of brain and spinal cord was reduced by more than two-thirds by BAL treatment. Further, the blood clearance of arsenic was increased. BAL treatment enhanced the elimination of arsenic in two ways: by decreasing the tissue-to-blood partitioning which mobilizes arsenic into the blood stream, and by increasing the clearance of arsenic.

Adipose Tissue

[Diagnostic values of BAL and TBLB in patients with interstitial lung diseases].

In order to evaluate the diagnostic values of bronchoalveolar lavage (BAL) and transbronchial lung biopsy (TBLB) in interstitial lung diseases, we made a retrospective study of a total of 43 patients all of whom underwent examinations for both conditions at the same time. In the BAL examination, fractional analyses of cell differential counts were applied to the first 30 ml lavage (FBAL-I, bronchial lavage) and to the following 50 ml aliquot of the second and third lavages (FBAL-II and -III, the latter supposedly alveolar lavage) lavages respectively. This technique revealed the unique characteristics of bronchial and alveolar inflammations in a separate data and this is assumed to be a precise reflection of the pathogenesis of interstitial lung diseases. The evaluation values (EV) were graded from 1 to 5 for both results according to the following diagnostic significance: EV1: not useful, EV2: normal or only slight changes from normal level, EV3: moderate changes although not diagnostic, EV4: quite compatible findings for diagnosis and EV5: definitely diagnostic. The 19 patients (43%) whose EV in TBLB were either 4 or 5 were diagnosed as having sarcoidosis (3), hypersensitivity pneumonitis (1), tuberculosis (2), pneumoconiosis (6), histiocytosis X (3), pulmonary infiltration with eosinophilia syndrome (PIE) (1), collagen vascular diseases (1), panbronchiolitis (1) or lymphagiomyomatosis (1). Thirteen of them had a BAL EV 4 which consisted of rather characteristic patterns of cell differential counts: thus their diagnoses were reconfirmed. In the other 4 patients with hypersensitivity pneumonitis, tuberculosis or PIE, the diagnoses were established by the findings of BAL EV 4 although the EV of TBLB were 3 or 2. These results indicate that BAL alone hardly establishes a diagnosis but it can support the TBLB findings and increase a diagnostic ability by 10% in total. Basophilic leukocytes, mast cells and Langerhans cells in BAL were rather non-specific, but the former two frequently appeared in allergic states and the latter two were often present in fibrotic lungs. Histiocytosis X is indicated if Langerhans cells are recovered from more than a small percentage of the total cell counts.

Adolescent

Clinical and subclinical alveolitis in collagen vascular diseases: contribution of alpha 2-macroglobulin levels in BAL fluid.

The probability that patients with collagen vascular diseases (CVD) will develop fibrosis is unpredictable. Since changes in bronchoalveolar lavage (BAL) cell data can be observed in CVD patients without evidence of lung involvement, we investigated whether the study of soluble components in BAL could help to distinguish CVD patients with lung involvement (n = 15) from those without pulmonary disease (n = 37). Our results demonstrate that the alveolitis observed in patients with overt lung involvement is associated with an increase of BAL alpha 2-macroglobulin (alpha 2-MA). In contrast, the BAL alpha 2-MA levels were found to be normal in CVD patients without evidence of pulmonary disease as well as in CVD patients with overt lung involvement treated with steroids. This was observed even in the presence of high neutrophil or lymphocyte counts in BAL. In conclusion, when neutrophils or lymphocytes accumulate in the lungs of CVD patients without evidence of lung damage, in the majority of patients this cell accumulation is not associated with an increase of BAL soluble components.

Adult

Amelioration by BAL (2,3-dimercapto-1-propanol) and DMPS (sodium 2,3-dimercapto-1-propanesulfonic acid) of arsenite developmental toxicity in mice.

Inorganic arsenic is embryotoxic and teratogenic in chicks, golden hamsters, mice, and rats. Certain dithiol chelators have been reported to protect against arsenite-induced lethality and to decrease arsenic body burden. The present study evaluated the influence of BAL (2,3-dimercapto-1-propanol) and DMPS (sodium 2,3-dimercapto-1-propanesulfonic acid), a water-soluble analogue of BAL, on arsenic-induced embryotoxic and teratogenic effects in the mouse. A series of four BAL or DMPS injections was administered sc to pregnant mice immediately after a single ip injection of 12 mg/kg of sodium arsenite given on Day 9 of gestation and at 24, 48, and 72 hr thereafter. Controls received sc corn oil with or without arsenite. Amelioration by BAL and DMPS of arsenite developmental toxicity was assessed at 15, 30, and 60 mg/kg/day, and 75, 150, and 300 mg/kg/day, respectively. BAL given following arsenite was not able to ameliorate the developmentally toxic effects of arsenite seen in mice, whereas treatment with DMPS at 150 and 300 mg/kg showed significant protective effects against arsenite embryotoxicity and teratogenicity. DMPS administration at 300 mg/kg also protected the dams against arsenite-induced maternal toxicity.

Animals

Comparison of Papanicolaou's stain with the Gomori methenamine silver (GMS) stain for the cytodiagnosis of Pneumocystis carinii in bronchoalveolar lavage (BAL) fluid.

The cytodiagnosis of Pneumocystis carinii (PC) in bronchoalveolar lavage (BAL) fluids has traditionally required a special stain such as Gomori's methenamine silver (GMS) stain. Recent reports indicate that identification of foamy alveolar casts (FACs) with Papanicolaou's (Pap) stain may provide a sensitive and less complicated way of making the diagnosis. To confirm these observations, results on a series of 318 BALs were reviewed. PC was identified on 65 (20%) specimens from 54 patients. Pap stains and GMS stains were positive on 56 (86%) of these BALs. Pap stains were positive on seven (11%) specimens that had negative GMS stains. PC was later confirmed on these specimens by other methods. Only two (3%) BALs had positive GMS stains and negative Pap stains. The results of this study confirm other reports that show that PC can be sensitively diagnosed with the Pap stain. The authors suggest that routine special stains for PC are unnecessary on BALs.

Bronchoalveolar Lavage Fluid

Hyaluronic acid (hyaluronan) in BAL fluid distinguishes farmers with allergic alveolitis from farmers with asymptomatic alveolitis.

Pulmonary function measurements, bronchoalveolar lavage (BAL), and analyses of precipitating antibodies in blood were performed in 12 farmers wtih no symptoms from the airways and 12 farmers who were admitted to the hospital due to acute symptoms of alveolitis (all nonsmokers). In addition, a bronchial methacholine provocation test was performed in the asymptomatic farmers. In 11 of the 12 symptomatic farmers but in none of the asymptomatic farmers, precipitating antibodies against one or more of the microorganisms which usually occur in a farmer's environment were found. In the farmers with symptomatic alveolitis, a restrictive impairment of pulmonary function was found, while pulmonary function was normal in all asymptomatic farmers. Findings in the BAL fluid showed increased concentrations of total cells, lymphocytes, and neutrophils and elevated levels of albumin, fibronectin, and angiotensin-converting enzyme in asymptomatic farmers compared with our own reference group. The same analyses in BAL fluid from the symptomatic farmers revealed a further increase in all parameters compared with the asymptomatic farmers. The BAL fluid from asymptomatic farmers had normal levels of hyaluronic acid (hyaluronan) and procollagen 3 N-terminal peptide, while these levels were significantly increased in the symptomatic group. We conclude that inflammation in the alveolar space and signs of activation of alveolar macrophages are present in farmers regardless of respiratory symptoms, although these findings are more pronounced in the presence of symptoms of acute alveolitis; however, the findings of impaired pulmonary function and the occurrence of precipitins and elevated levels of hyaluronic acid and procollagen 3 N-terminal peptide in BAL fluid were exclusively found in the farmers with airways symptoms. We postulate the hyaluronic acid, due to its pronounced ability to immobilize water, may be of importance in the development of the pulmonary function impairment observed in farmer's lung disease.

Acute Disease

In vivo evaluation of bronchoalveolar lavage (BAL) fluid in atopic bronchial asthma, chronic bronchitis and sarcoidosis patients.

The intensity of inflammatory response was evaluated in skin test on guinea pig using bronchoalveolar lavage (BAL) fluid obtained from patients with some diseases of the respiratory tract. The results of skin test were verified with activities of proteases in BAL fluid. The study was performed on 24 patients with atopic bronchial asthma, 21 with chronic bronchitis, 13 with sarcoidosis (II phase) and 18 control subjects. All patients were undergoing fiberoptic bronchoscopies and BAL fluid was obtained. The results of skin test on guinea pig using BAL fluid were correlated with the activities of acid and neutral proteases. The highest activity of proteases and intensity of skin reactions were noted in patients with atopic bronchial asthma and sarcoidosis. Authors suggest that the skin test on guinea pig with BAL fluid may be useful tool for total evaluation of inflammatory response in patients with atopic bronchial asthma, chronic bronchitis and sarcoidosis.

Adult

[IgE-Fc receptor expressions on bronchoalveolar lavage (BAL) cells in a patient with eosinophilic pneumonia accompanied with bronchial asthma--the effect of ketotifen on their expression].

In this study, IgE-Fc receptor expressions on each type of BAL cells (lymphocytes, macrophages and eosinophils) in a patients with eosinophilic pneumonia accompanied by bronchial asthma were examined by indirect immunofluorescent method using monoclonal antibody (H107). BAL cell findings showed marked increases of total BAL cell counts and eosinophils, an increased number of lymphocytes and the presence of basophilic cells. These results match those of our previous report. Furthermore, IgE-Fc receptor expressions on lymphocytes, macrophages and eosinophils were markedly increased as compared to those in the peripheral blood and normal control subjects. These findings suggest that IgE production in the lung plays the main role in the pathogenesis of eosinophilic pneumonia and bronchial asthma. On the other hand, during ketotifen administration, decreases of peripheral blood eosinophilia, of eosinophilia in the sputum and of serum IgE level and an improvement in chest X-ray findings were observed. Furthermore not only normalization of total BAL cell counts but also decreases of IgE-Fc receptor expressions on BAL cells were observed.

Adult

[Bronchoalveolar lavage (BAL)--a diagnostic method in chronic nonspecific bronchopulmonary diseases in childhood. 1. Implementation and tolerance].

While the effectiveness of bronchoalveolar lavage (BAL) in pediatric therapy has been recognized for years, there are hardly any data on its usefulness in diagnosis, its performance and compatibility in childhood. After 123 bronchological investigations including BAL on 118 children in the age of three months through 16 years, with lavage fluid amounts between 40 and 160 ml, febrile reactions occurred in 17 cases (= 13.8%), i.e. no more frequent than in a group of 100 children bronchologically investigated but without BAL. After blood gas analyses, ECG examinations, blood pressure readings the compatibility of BAL turned out to be good in children of every age. Bronchography pictures after BAL were not reduced in quality, atelectasis formation occurred rarely.

Adolescent

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.

Adult