A novel covalent enzyme-linked immunoassay (CELIA) for simultaneously measuring free and immune complex bound antibodies of defined specificity. I. Application to naturally occurring antipolyamine antibodies in human sera.

A simple covalent enzyme-linked immunoassay procedure (CELIA) is described for the routine determination of free and immune complex-bound antibodies in sera. Assays for the latter could not have been performed by adsorption ELISA due to the high ionic strength of the reassociating buffer. For the measurement in human sera of free naturally occurring IgG and IgM antibody directed against the hapten spermine, polycarboxystyrene microtiter plates with covalently coupled spermine were used. For the determination of immune complex-bound antipolyamine IgG and IgM antibody titers, serum was first dissociated at pH 2.3 in tubes and then reassociated at pH 8.1 in the wells of a microtiter plate containing covalently bound spermine. The reactivity of anti-spermine antibodies was increased from 2- to 13-fold after dissociation and reassociation compared to that of non-dissociated area. The apparent reaction constant (Rapp.) of free IgG antibodies to spermine in the sera of 19 bronchopulmonary patients with cancer differed significantly from Rapp. values of IgG antibodies having this specificity in ten other patients with non-malignant disease.

[Analysis by broncho-alveolar lavage of the pulmonary uptake of gallium 67 in idiopathic diffuse interstitial fibrosis and sarcoidosis]. / Analyse par lavage broncho-alvéolaire de la fixation pulmonaire du gallium 67 au cours des fibroses interstitielles diffuses idiopathiques et des sarcoïdoses.

The pulmonary fixation of Gallium 67 has been studied by broncho-alveolar lavage (LBA) in a group of 10 cases of stage II pulmonary sarcoid, proven histologically, and in a group of 11 cases of idiopathic diffuse interstitial fibrosis (FID). The pulmonary activity of Gallium 67 was measured in vitro in a quantitative manner on a centrifuged clump and a supernatant of LBA as well as on the serum and in vivo in a semi-quantitative manner by the Line index. The alveolar Gallium activity has been correlated with different cytological and biochemical parameters of the lavage liquid. There was a narrow correlation in the serum between the Gallium 67 activity and the level of circulating transferrin. The fixation of Gallium in the cellular clump of LBA depends on the number of macrophages present in the clump and the total available radioactivity. The macrophages of the sarcoid and fibrosis cases picked up the gallium in a similar fashion, with an affinity all the more important as the total available radioactivity was weak. The radioactivity of the supernatant of the lavage liquid was quantitatively important and narrowly tied to the diffusible proteins present in the lavage. There was no correlation between the semi-quantitative Line index and the cytological or biochemical data in the fibrosis cases. In the sarcoid cases there was an inverse correlation with the polymorphonuclear leukocyte count. The semi-quantitative index was only correlated with alveolar gallium fixation level in the fibrosis group.(ABSTRACT TRUNCATED AT 250 WORDS)

Lactic dehydrogenase isoenzyme electrophoretic patterns in the diagnosis of pleural effusion.

The relative value of each lactic dehydrogenase isoenzyme (iso LDH) was measured by electrophoretic separation in the serum and the pleural fluid of 100 patients. In each case, the cause of the pleural effusion was known. Two types of LDH isoenzyme pattern were found in the serum: a normal type with a low value of LDH 5 and an abnormal type with a high value of LDH 5. This high LDH-5 level is due to an impaired liver function. In the pleural fluid, the electrophoretic patterns of five LDH isoenzymes were found by computerized processing. During congestive heart failure (28 cases) the electrophoretic pattern of the LDH isoenzymes was always similar in the serum and in the pleural fluid (transudative pleural effusion). During thoracic empyema, the relative values of the isoenzymes in the pleural fluid were regularly increasing from LDH 1 to LDH 5. In this situation, the evaluation of LDH 5 appeared to emanate from the increased granulocytes in the pleural fluid. In 22 inflammatory pleural effusions, the relative values of the five isoenzymes were equal. During malignant effusions (35 cases) a high level of LDH 5 was found in 21 patients. LDH 5 is known to be secreted by malignant tissue, and the authors confirmed it by finding a high level of LDH 5 in biopsy specimens of patients with mesothelioma or epidermoid lung cancer (7 cases). Conversely, the level of LDH 5 was low in biopsy specimens from normal lung tissue or benign inflammatory pleuritis (6 cases). Among the 14 patients with low levels of LDH 5 in the pleural fluid during malignant pleural effusion, the authors found the malignant lymphomas (three cases) and the small cell lung carcinoma (five cases). In these cases, the low level of LDH 5 was in agreement with the result of a low level of LDH 5 found in the biopsy of a metastatic liver localization of a small cell lung carcinoma. So the electrophoretic determination of LDH isoenzymes pattern in pleural fluid is a sensitive tool for the management of pleural effusion.

Protective effects of dietary calcium and magnesium on platelet function and atherosclerosis in rabbits fed saturated fat.

Male rabbits were fed for 6 months a purified diet rich in fat (45% of calories), mostly saturated (butter) containing per 100 g either 354 mg of calcium and 46 mg of magnesium (group I), 948 mg of calcium and 46 mg of magnesium (group II), or 354 mg of calcium and 356 mg of magnesium (group III). In group II, fed the additional calcium, significant (P less than 0.001) changes were observed; i.e. prolongation of clotting time (platelet clotting activity), decrease in platelet aggregation to thrombin, and in the concentration of the plasma total cholesterol, with a less significant (P less than 0.05) decrease in plasma triglycerides. A significant (P less than 0.01) decrease in the ratio monoemes/18:2 in the plasma cholesterol esters but not in the plasma or platelet phospholipids was also found in these rabbits. In relation to their lower activity, platelets from group II exhibited a significant increase in 22:4n-6 (phospholipids) and in the ratio cholesterol/phospholipids, both significantly correlated with the platelet function tests. In this same group, the excretion of fecal lipids and saturated fatty acid 18:0 was increased by 4.3-fold and 7.6-fold, respectively. The severity of atherosclerosis and the accumulation of cholesterol in the aorta were significantly lower in group II, while the plasma level of calcium was similar to that of group I. In group III, fed additional magnesium, results were similar to those of group II, but less significant.

An automatic continuous flow method for the determination of antipolyamine antibodies in human sera.

Latex particles with covalently bound polyamines were used to detect antipolyamine antibodies in human sera in manual and automated nephelometric assays. There was good correlation between the two assays though in the former aggregate size was measured after 24 h incubation whereas in the latter monomer loss was determined after 1 h.

Evidence for natural antibodies (IgG) to polyamines in human sera.

Human sera contain IgGs that react with latex-putrescine spheres. Identification of the IgGs has been achieved by polyacrylamide gel electrophoresis and their reaction with peroxidase-labeled antihuman gamma-chain-specific antibodies. The inhibition of IgG fixation to latex-putrescine spheres by free spermine and putrescine provides evidence that these IgGs are specific for polyamines.

[Rapid and automatic acquisition and transmission of acid-base equilibrium parameters in a clinical biology laboratory (author's transl)]. / L'équilibre acido-basique. Acquisition et transmission rapides et automatiques de ses paramètres dans un laboratoire de biologie clinique.

The "pH-blood gas" department of our clinical biology laboratory was organized to meet rapidly and in the most reliable conditions possible the numerous and urgent analytical demands that had been confronting us. The structure was set up, therefore, in order to accelerate the speed at which blood samples could be transferred and make it possible to quickly and reliably obtain and transmit the results of the analyses. These necessities were satisfied thanks to the installation of a pneumatic tube allowing the rapid transfer of syringes, the utilization of automatic analysers, and the installation of a data processing system.

[Colorimetric determination of calcium in serum using methylthymol blue]. / Determination du calcium serique par une technique utilisant le bleu demethylthymol

A simple, specific and rapid colorimetric method for the determination of calcium concentration and serum using methylthymol blue is described. The reagent given can be used both in a manual method and in an automatic method using an auto-analyser without dialysis. The results obtained by the automatic method are compared to those found by the manual method and those obtained by atomic absorption.

[Value of phosphohexose isomerase (PHI) determination in myocardial infarct]. / Intérêt du dosage de la phosphohexose-isomérase (PHI) dans linfarctus du myocarde

The biochemical diagnosis of myocardial infarction is based mainly on the enzyme levels of glutamino-oxalic transaminase (SGOT), lactic dehydrogenase (LDH), and creatine-phosphokinase (CPK). The authors have compared the level of phospho-hexo-isomerase (PHI) with the levels of the three other enzymes mentioned above in 20 cases of recent myocardial necrosis. From their study it appears that PHI activity follows essentially the same pattern as that of CPK, and has no particular advantage; indeed, it is limited by its later appearance and its lower maximum level compared with CPK. Finally, because of its lack of specificity, the practical importance of PHI measurement in the clinical diagnosis of myocardial infarction is small.