Behaviour of leukocyte membrane fluidity in presence of anaesthetic drugs. Comparison between allergic patients and control subjects.

1. In this study we compared the effects of two anaesthetic drugs on the leukocyte membrane fluidity in allergic patients versus control subjects. 2. Fluidity was assessed by means of the fluorescence polarization technique. 3. We report that the treatment of the whole leukocytes with thiopental or pancuronium enhanced membrane fluidity in the allergic group as well as in the control one, but the effect was more pronounced in allergics. 4. This finding suggests a different biophysical behaviour of the leukocyte membrane towards anaesthetic drugs in allergic diseases. 5. This agrees with the hypothesis of the existence of an intrinsic abnormality in allergic cells, expressed as an initial hyperreactive state.

Impaired G-proteins and cyclic nucleotide phosphodiesterase activity in T-lymphocytes from patients with sarcoidosis.

Among the various immune abnormalities which characterize active sarcoidosis, a low proliferative response of peripheral blood lymphocytes to mitogenic lectins has long been observed. Since membrane-associated G-proteins are very likely to be crucial elements in lectin signal transduction, we investigated the binding of 5'-guanylylimidodiphosphate (GppNHp), a non hydrolyzable GTP analogue, to blood total lymphocyte membranes and to blood T-lymphocyte membranes from patients with active sarcoidosis, and from healthy control subjects. GppNHp binding was markedly decreased in peripheral cells from patients with sarcoidosis as compared to controls, suggesting the occurrence of a defect at the level of G-protein(s). A further characterization of G-proteins in these cells by means of ADP-ribose-labelling in the presence of bacterial toxins brought forward a significant decrease in the labelling of a 40 kDa protein, the major pertussis toxin substrate, in membranes from sarcoid patients, while the labelling of the major 44 kDa cholera toxin substrate proved to be unchanged with respect to control membranes. It is hypothesized that, in sarcoid lymphocytes, a defect in the negative control of adenylate cyclase mediated by the inhibitory G-protein Gi, prevents the lowering of cAMP necessary to normal mitogenic response of blood lymphocytes to stimulation. cAMP degradation by the specialized enzyme phosphodiesterase constitutes another critical step in the control of cAMP levels. Both cAMP and cGMP phosphodiesterase activities were found decreased in blood total lymphocyte preparations from sarcoid patients. With purified T-cells, although the mean cAMP and cGMP phosphodiesterase activities from sarcoid patients were found more markedly decreased with respect to healthy donors, only the decrease in cGMP phosphodiesterase was found statistically significant. The role these defects in cyclic nucleotide degradation potentially play in the disturbance of blood lymphocytes response associated with sarcoidosis is discussed.

Increased membrane fluidity in blood leukocytes from allergic subjects. A possible role for leukotriene B4.

Modifications of lipid metabolism and increased plasma membrane fluidity may occur during cell activation and occur in leukocytes of patients with allergic disease. Using a fluorescent probe (TMA-DPH) and a polarization technique, we studied the variations of membrane fluidity in blood leukocytes from allergic and normal subjects. The anisotropic coefficient, which is inversely related to the rotational motion of the probe in membrane phospholipids, was significantly higher in normal subjects than in allergic ones. This result implies an increased membrane fluidity in leukocytes from allergic patients. No correlation could be observed with the etiology or the severity of allergic disease, nor with cell preparation composition or serum IgE level. Such a modification of membrane fluidity has been reproduced after incubation of leukocytes from normal subjects with leukotriene B4, but not with cells from allergic patients. Establishing the mechanism of these differences requires further investigations.

Cyclic nucleotide phosphodiesterases and methyltransferases in purified lymphocytes, monocytes and polymorphonuclear leucocytes from healthy donors and asthmatic patients.

Cyclic nucleotide phosphodiesterase and phospholipid N-methyl-transferase activities were simultaneously measured in purified polymorphonuclear cell-, mononuclear cell-, lymphocyte- and monocyte-homogenates from control subjects, from patients with atopic asthma and from patients with non-atopic asthma. Whereas cyclic AMP and cyclic GMP phosphodiesterase activities were found to be about 10-fold lower in polymorphonuclear than in mononuclear cells, phospholipid N-methyltransferase proved to be rather similar in each cell type from control donors. Cyclic AMP phosphodiesterase and phospholipid N-methyltransferase were significantly decreased in polymorphonuclear cells and monocytes from asthmatic patients compared with the control group while cyclic GMP phosphodiesterase was significantly impaired only in the monocyte subpopulation.

[Karyotype study of blood lymphocytes in sarcoidosis]. / Etude caryotypique des lymphocytes sanguins dans la sarcoïdose.

Among 31 patients presenting with pulmonary sarcoidosis, two had abnormalities of their blood lymphocyte karyotypes. The karyotype of the first patient showed an initially high percentage of non-specifically broken chromosomes. The second patient, who had been treated with azathioprine (AZ) one year previously, had an apparently balanced translocation 46 XX, t (11; 11) (p 12, p 14) in blood T lymphocytes but not in skin fibroblast culture. Various hypotheses can be discussed to explain this translocation: a direct toxic effect of AZ or a genomic abnormality depending upon sarcoidosis and possibly revealed by AZ. It is important to note that this translocation concerned a region of the short arm of chromosome 11, where Harvey ras I and parathormone genes have been located.

Evaluation of histamine release following intravenous injection of ionic and nonionic contrast media.

To evaluate histamine release (HR) following injection of radiocontrast media and to test the predictive value of peak-expiratory-flow rate (PEFR) in detecting high-risk patients, the authors performed in two series 90 intravenous pyelograms (IVPs). In the first group, HR was measured by a fluorometric method after injection of Hexabrix (25 patients) and Iopamiron (25 patients). In the second group, HR measured by a radioimmunoassay, and PEFR measured by a peak flowmeter were investigated after injection of Hexabrix (10 patients), Telebrix (10 patients), Omnipaque (10 patients) and Iopamiron (10 patients). Histamine release in groups 1 and 2, and PEFR in group 2, were not significantly modified by the injection of each radiocontrast media. For the four patients (two per group) who experienced minor allergic side effects, the levels of HR and PEFR were always within the normal ranges.

[A simple test for the detection of respiratory allergy to current pneumallergens]. / Un test simple de dépistage de l'allergie respiratoire aux pneumallergènes courants.

Phadiatop is a new in vitro test used for screening patients with allergic respiratory diseases. This test, based on the RAST procedure, is performed with a paper disc to which a balanced mixture of relevant inhalant allergens has been coupled. One hundred and one adults ranging in age from 18 years to 73 years and 38 children (4 months to 15 years) were studied. We found a good correlation between this new test and total IgE (adults: 74.3 p. 100, children: 68.4 p. 100), specific IgE (adults: 91.1 p. 100, children: 100 p. 100) and skin test (adults: 79.2 p. 100, children: 97.1 p. 100). There was a high correlation between clinical atopy assessment and the result of Phadiatop, its efficiency was 83 p. 100 in adults and 97 p. 100 in children.

[Comparative study of various technics for measuring serum IgE levels]. / Etude comparative de différentes techniques de dosage des IgE sériques spécifiques.

In this work we report the results of a comparative study of different techniques to measure specific IgE in sera. Three methods were used, Phadebas RAST, Phadezym RAST (PHARMACIA) and allergen-specific FAST+ test (3M DIAGNOSTICS): 127 sera were tested with several pneumoallergens (housedust, Dermatophagoides pteronyssinus, Dactylis glomerata pollen, cat and dog hair). Analysis of the results shows the following correlations: Phadebas and Phadezym RAST 92%; Phadebas RAST and FAST+ 88%; Phadezym RAST and FAST+ 67%; Furthermore, the responses vary according to the allergen. The FAST+ system seems to be more sensitive than the RAST. The RAST must remain the reference method, but it should always be remembered that the different techniques for measurement of specific IgE have their own fields of application.

Phosphatidylethanolamine methyltransferase and cAMP, cGMP phosphodiesterases in lymphocytes and monocytes in sarcoidosis.

Among the various hypotheses proposed to explain immune cell defect in sarcoidosis, we examined thoroughly that of Faguet who described abnormalities of signal transmission at lymphocyte membrane level. Phosphatidylethanolamine methyltransferase and cAMP cGMP phosphodiesterases were studied in blood lymphocytes and monocytes from 8 subjects with sarcoidosis disease. Phosphatidylethanolamine methyltransferase (PMT1) plays an important regulatory role in membrane signal transmission. cAMP and cGMP phosphodiesterases (PDE) regulate cytoplasmic cyclic nucleotide levels and so participate in the modulation of the cell cycle. We observed a decreased PMT1 activity in lymphocytes and monocytes and a decreased cAMP and cGMP PDE activities in monocytes. It is not now possible to say if these abnormalities are primary or secondary. Whatever the origin of this dysfunctioning, these results evoke simultaneous disturbances of membrane signal transmission and cell cycle in monocytes and membrane abnormalities in lymphocytes. These abnormalities could explain some immune cell defects in sarcoidosis disease.

[Methylation of membrane phospholipids of alveolar macrophages in pulmonary sarcoidosis and diffuse interstitial pulmonary fibrosis]. / La méthylation des phospholipides membranaires du macrophage alvéolaire dans la sarcoïdose pulmonaire et les fibroses pulmonaires interstitielles diffuses.

Alveolar macrophage plays an important role in alveolitis pathogenesis and lung fibrosis process. Macrophages exhibit plasmic membrane biochemical modifications during cell activation. Phospholipid methylation is involved during plasmic membrane stimulation. Phosphatidylethanolamine methyltransferase I was measured in alveolar macrophage membrane from normal subjects and patients presenting with pulmonary sarcoidosis or interstitial lung fibrosis. We observed increased enzyme activity among sarcoidosis subjects with high intensity lung alveolitis and normal subjects receiving immunostimulating treatment (RU 41740). The role of membrane phospholipidic modifications in granulomatous process is discussed.

Alveolar macrophage membrane phospholipid methylation in sarcoidosis and hypersensitivity pneumonitis.

Methylation of phospholipids seems to be an essential step in the recognition and transduction of regulatory signals by eukaryotic cells. Phosphatidylethanolamine methylation was compared in alveolar macrophage membrane from patients presenting with pulmonary sarcoidosis or hypersensitivity pneumonitis and control subjects. Phosphatidylethanolamine methyltransferase (PMT1) activity was determined by various measures of incorporation of tritiated methyl group from (3H) S-adenosyl-L-methionine in membrane phospholipids. Tritiated methyl group incorporation in macrophage membrane was higher in some patients presenting with sarcoidosis or hypersensitivity pneumonitis, than in controls. PMT1 activity was found to be higher in sarcoidosis patients with a positive gallium lung scan. As lipids play an important role during macrophage activation and cell interaction, although a wide heterogeneity was observed in PMT1 activity, increased membrane phospholipid methylation seems to be an important feature in pulmonary diseases where macrophages are involved.

Azathioprine treatment of chronic pulmonary sarcoidosis.

The object of this study was to evaluate the effectiveness of the immuno-suppressor Azathioprine (AZ) on chronic and severe pulmonary sarcoidosis, with a persistent activity resistant to prolonged corticotherapy. The study was done on 10 patients (4 women, 6 men) afflicted by an histologically proven and chronic sarcoidosis, resistant to steroid treatment. The treatment consisted of a daily oral intake of 150 mg of AZ for six months. Its effectiveness was evaluated before and after treatment, in comparison with a control group and a steroid-treated group of sarcoid patients. Serologic and alveolar functional and immuno-biologic tests were performed in 8 cases according to the activity criteria defined at the IXth International Congress on sarcoidosis. No clinical or hematological side effects were observed; a clear and prolonged radiological and clinical amelioration was observed in 7 out of 10 cases and in 3 cases a restoration of sensibility to the tuberculin skin test; in the 8 cases a significant improvement (p less than 0.01) was noted after the sixth month of treatment only in the alveolar fluid in the following parameters: ACE, all the proteins studied and the percentage of lymphocytes.

[Respiratory function and alveolar biological changes under the effect of CDP-choline in pulmonary interstitial pathology: pulmonary fibrosis and sarcoidosis]. / Etude des modifications fonctionnelles respiratoires et biologiques alvéolaires sous l'effet de la CDP-choline dans le cadre de la pathologie interstitielle pulmonaire: fibroses pulmonaires et sarcoïdoses.

Various anomalies of pulmonary surfactant have been described in relation to acute respiratory distress syndromes, hypersensitivity lung disease and pulmonary sarcoidosis. Phosphatidylcholine (PC) is the essential phospholipid component of pulmonary surfactant. Cytidine diphosphocholine (CDP-choline) is an essential intermediary in the biosynthesis of PC. The authors studied two groups of patients: one group consisted of diffuse interstitial pulmonary fibrosis and the other consisted of pulmonary sarcoidosis with parenchymal involvement. They observed quantitative and qualitative abnormalities of the phospholipid fractions of surfactant and more particularly of PC. The finding of a marked decrease in this phospholipid, especially in the cases of pulmonary fibrosis, justified the study of the therapeutic effects of CDP-choline. After one month of treatment with this substance, at a dose of 1 g I.M. per day, the PC fraction had returned to normal and, at the same time, there was an improvement in the PaO2 at rest and after exercise. Long term administration of CDP-choline appears to be valuable in the maintenance of the phospholipid equilibrium of pulmonary surfactant and in the improvement of the quality of alveolar gas exchange.

[Bronchoalveolar lavage data in localized pulmonary tuberculosis]. / Données du lavage broncho-alvéolaire dans la tuberculose pulmonaire localisée.

Twenty-two patients with radiologically localised pulmonary tuberculosis underwent one or more broncho-alveolar lavages: 10 patients had a single lavage in the disease area, 11 had two lavages (1 in a healthy zone and 1 in the affected zone) and 1 patient had a triple lavage. The laboratory analysis of the fluid revealed that the tuberculosis was accompanied by an elevation of the various proteins assayed and by an alteration in the cytology (lymphocytosis, neutrophilia or mixed) in the radiologically involved zone. In contrast,the healthy zone revealed normal values except in 2 cases which presented a lymphocytosis. The type of cytological abnormality does not appear to have any prognostic value, but is probably related to different pathophysiological mechanisms.

[Pulmonary sarcoidosis. Study of the distribution of active alveolitis assessed by comparison of data from radiological examination, gallium 67 scintigraphy and double bronchoalveolar lavage]. / Sarcoïdoses pulmonaires. Etude de la répartition de l'alvéolite active appréciée par comparaison des données de l'examen radiologique, de la scintigraphie au gallium 67 et de doubles lavages broncho-alvéolaires.

Abnormal uptake of gallium in alveolar structures, raised lymphocyte levels, ECA and, to a lesser degree, proteins in alveolar washings are the principal currently known criteria used to recognise cases of sarcoidosis with persistent immunological activity. The contribution of data concerning broncho-alveolar washings (BAW) in the context of this diagnostic assessment has never been the subject of a critical evaluation. It would nevertheless seem that the distribution of sarcoidosis lesions is not always diffuse and homogeneous, leading to the possibility of errors of assessment in this technique. The aim of the present study was to assess the true existence, degree and consequences of this risk. It was based upon the study of 41 cases of active pulmonary sarcoidosis. Search for homogeneity and heterogeneity between lesions and active alveolitis involved precise radiological study, gallium isotope scan with determination of the index of uptake as well as the quality of the latter, and double BAW carried out in the most radioactive and least radioactive regions respectively. Comparative study of the results obtained showed that zones of active alveolitis were distributed unevenly and heterogeneously in more than half of the patients (71% for gallium and 54% for BAW). This would explain the fact that in 14 cases (34%) criteria of alveolar activity were present in only one of the two specimens of washings. Identification of the zones which should be included in BAW cannot be determined accurately by radiological analysis, regardless of the degree and distribution of radiological abnormalities.(ABSTRACT TRUNCATED AT 250 WORDS)

[Membrane phosphatidylethanolamine methylase in blood leukocytes and alveolar macrophages of asthmatic patients]. / Etude de la phosphatidyl ethanolamine méthylase (PEMT) membranaire sur les leucocytes sanguins et les macrophages alvéolaires chez le sujet asthmatique.

Phosphatidyl ethanolamine methylase (PEMT) is an enzyme involved in the methylation of membrane phospholipids which plays a very important role in the modulation of the activity of the beta-receptors and the production of phosphatidylcholine, substrate of phospholipase A2. This report describes a study of PEMT on the membranes of blood leucocytes and alveolar macrophages obtained by bronchoalveolar washings in different types of asthma: allergic, intrinsic and occupational. This was accompanied by parallel study of respiratory function tests and the level of bronchial reactivity to carbachol in asthma sufferers as well as other alveolar biological parameters: phospholipid fractions of surfactant, angiotensin conversion enzyme and protein/albumin ratio. The authors found a significant increase (in comparison with controls) in PEMT activity both in macrophages and leucocytes in cases of intrinsic asthma. However, there was no correlation between the level of activity of the enzyme and the degree of bronchial hyper-reactivity. There was no change in alveolar phospholipid environment. In the light of these findings, the role of membrane activation of the alveolar macrophage is discussed in the physiopathology of intrinsic asthma.

Activity of cyclic AMP phosphodiesterase and methyltransferases in leukocyte membranes from allergic patients.

Cyclic AMP metabolism and methylation of phospholipids are central events which occur at the membrane level. Since a dysfunction of cell membranes seems to characterize some allergic diseases, we investigated cyclic AMP phosphodiesterase and methyltransferase activities in leukocyte membrane fractions obtained from healthy volunteers and from allergic patients. The allergic group presented a significantly decreased methyltransferase activity compared with a control group, whereas cyclic AMP phosphodiesterase and noradrenaline (NA)-stimulated methyltransferase were found to be increased with respect to the control group. A significant correlation has been found between cyclic AMP phosphodiesterase and NA-stimulated methyltransferase with both control and allergic subjects, which suggests close relationships between these two enzymes within the cell membrane.

[Modification of the Friedland-Silverstein technic for the determination of the activity of angiotensin I converting enzyme]. / Modification de la technique de Friedland et Silverstein pour la détermination de l'activité de l'enzyme de conversion de l'angiotensine I.

The authors studied the parameters of Friedland and Silverstein's technique for measuring the activity of the angiotensin I conversion enzyme and found a number of anomalies. The technique was modified by correcting the wide pH difference between the pH of the reagent and the pH of the reference solution. This modification made the technique optimal for the measurement of the enzyme activity by allowing a 20 to 40 per cent increase in this activity and a 15% increase in the positivity in cases of clinical sarcoidosis. The application of this modified test to the determination of the activity of the angiotension I conversion enzyme in control subjects, patients with sarcoidosis and patients with order respiratory diseases gave the following mean values, respectively: 39 +/- 10 IU; 81 +/- 23 IU; 41.5 +/- 2.5 IU in the serum and 0.2 IU; 1.1 +/- 0.5 IU; 0.2 +/- 0.1 IU in bronchoalveolar lavage fluid. The increase in enzyme activity in sarcoidosis is found in the serum in 75% of cases and in bronchoalveolar lavage fluid in 85% of cases. This modified technique can be applied in routine practice and in research.