Analysis of pleural effusions using automated flow cytometry.

Flow cytometry allows rapid and accurate analysis of the deoxyribonucleic acid (DNA) content of a large number of cells. In solid tumors, the presence of aneuploidy has been shown to correlate wall with the presence of neoplastic cells. Both cytologic examination and DNA analysis by flow cytometry were performed on pleural effusions from 33 patients. Results of the two examinations were in agreement in 10 of 12 malignant pleural effusion (two false-negatives) and in 20 of 21 benign effusions. One patient with cirrhosis, ascites and Nocardia pneumonia had hypodiploid cells (false-positive) in the pleural fluid. All patients who had a malignancy, but whose pleural effusion proved to be due to a benign cause, had cells with normal DNA content in their pleural effusion. DNA analysis using flow cytometry can be rapidly performed and is highly specific and sensitive. The finding of hyperdiploid cells is highly suggestive of malignancy.

The pathogenesis of the low pleural fluid pH in esophageal rupture.

To determine the possible mechanisms responsible for the low pH pleural effusion associated with esophageal rupture we evaluated the following possibilities: (1) gastric acid reflux, (2) bacterial metabolism, and (3) leukocyte metabolism. Neither elimination of gastric hydrogen ion contribution by distal esophageal ligation nor elimination of bacteria with antibiotics prevented the progressive fall in pleural fluid pH after esophageal rupture. Only elimination of polymorphonuclear leukocytes from the pleural space by rendering animals leukopenic with nitrogen mustard, prevented a low pH effusion after esophageal rupture. It appears that pleural fluid leukocyte metabolism is primarily responsible for the low pH effusion associated with esophageal rupture.

Inflammatory responses in experimental tuberculosis pleurisy.

A model of tuberculous pleurisy in New Zealand white rabbits was developed to describe the sequential cellular and biochemical changes in pleural fluid. Bacille Calmette-Guérin (BCG) in 4 X 10(7) colony-forming units was introduced into the right pleural space of rabbits previously sensitized by intradermal BCG. Pleural fluid was obtained via serial thoracenteses. A normal-pH, normal-glucose, exudative effusion was seen through 144 hours. Polymorphonuclear leukocytes were the first cells to respond to the introduction of tubercle bacilli in the pleural space; they remained the predominant cell for the first 24 hours and were followed by macrophages, which peaked at 96 hours, and then by lymphocytes. Numerous granulomata were observed on both the visceral and parietal pleura ten days following intrapleural instillation of BCG. We propose that the polymorphonuclear leukocyte influx is not a nonspecific response to pleural injury and that such a leukocyte response, either itself or through its interaction with the macrophage, plays a role in host defense mechanisms against the tubercle bacilli.

Gas-liquid chromatography-frequency pulse-modulated electron-capture detection in the diagnosis of infectious diseases.

The extremely sensitive and selective gas-liquid chromatography-frequency pulsed-modulated electron-capture detection (GLC-FP-ECD) procedure has been applied to the diagnosis of bacterial, fungal, viral, rickettsial, and parasitic diseases by the examination of various body fluids, effusions and exudates, and excretion products. Carboxylic acid and alcohol, hydroxy acid, and amine product profiles of microbial or host-response origin, have been used to establish specific etiologies, these profiles are reproducible, and can be used to aid in the diagnosis of infections. In addition, we have used the GLC-FP-ECD procedure to analyze microbial metabolic products in vitro and to provide data for identification and classification. We also explored computer time-sharing for data analysis, profile library comparison, and eventual profile matching for diagnosis.

[Clinical effects of human lymphoblastoid interferon in patients with hematologic neoplasms].

Sixteen patients with hematologic neoplasms were treated with Human Lymphoblastoid Interferon (HL-BI) derived from Namalwa cell line. They were 6 multiple myeloma, 8 acute leukemia and 2 malignant lymphoma patients. All patients were previously treated with anticancer agents except one case with multiple myeloma. HLBI, 3.0 X 10(6) IU/day, was daily administered by intramuscular injection at least for 4 weeks. Three of 6 multiple myeloma responded to HLBI with a decrease of more than 25% in serum myeloma protein level. A case with pleural effusion due to massive infiltration of myeloma cells treated with intrathoratic administration of HLBI, in whom complete disappearance of pleural effusion was recognized. On the other hand, no patients with acute leukemia and malignant lymphoma responded except one case with acute lymphocytic leukemia, in which bone marrow lymphoblasts decreased transiently. Fever episodes, 13 of 16 cases, were more frequently seen but were manageable. Transient leukocytopenia and thrombocytopenia were also observed in 4 and 7 of 8 cases, respectively. No anaphylactoid reaction was seen. Thus, HLBI was expected useful in the clinical management of multiple myeloma.

[Pharmacokinetics and pleural reaction of doxycycline after intrapleural administration].

Tetracycline has been used for the local treatment of malignant pleurisy ever since of Robinson (1972) reported its effectiveness. In this report, we try to elucidate possible mechanisms of action of this drug from the viewpoints of (1) pharmacokinetic analysis following intrapleural administration, and (2) pleural reaction. The results were as follows: (1) In 5 patients with malignant pleural effusion, 500 mg of doxycycline was injected into the pleural cavity and pharmacokinetic analysis was performed. The clearance curve of doxycycline in pleural fluid was described by either a one-compartment model or a two-compartment model. The mean half life of slow space was 33.3 hr, which suggested delayed excretion of this drug from pleural space. (2) The direct effect of doxycycline on pleura was studied in rabbits. At a dose of 10 mg per kg of body weight, the mesothelial cells became cuboid and contained vacuoles in their cytoplasma. With increasing dose up to 40 mg per kg, the changes in mesothelial cells were enhanced. In submesothelial tissue, edema and cellular infiltration were noticed. Three weeks later, normalization of mesothelium was observed, being followed by connective tissue proliferation. These findings may support the efficacy of tetracycline for local treatment of malignant pleurisy.

DNA flow cytometry of pleural effusions. Comparison with pathology for the diagnosis of malignancy.

A study was undertaken to determine the potential value of DNA flow cytometry (FCM) for the diagnosis of malignancy in pleural effusions and to compare its results with those of traditional cytomorphology. Forty-one pleural fluids from 37 patients were evaluated by DNA FCM and routine cytologic techniques. Of the 41 pleural fluids, 29 (70.7%) demonstrated an abnormal DNA content by FCM. Two of the 29 pleural fluids had been originally diagnosed as benign by cytology. Cytologic review of these two cases showed no abnormal cells; therefore, there were no false-negative cases based on cytology. In 12 (29.3%) of the 41 fluids, DNA FCM and cytologic evaluation both indicated benign processes. Our preliminary observations indicate that FCM is an accurate and reliable technique that may be of aid in the diagnosis of malignant effusions. The technique may prove to be of special value in the differential diagnosis of reactive mesothelial cells versus malignant mesothelioma as well as for following patients who receive chemotherapy for malignant pleural effusions. DNA FCM may also complement cytomorphologic diagnoses in other serous, exfoliative or aspiration material.

Pleurisy in rheumatoid arthritis.

In 157 women and 77 men with rheumatoid arthritis, observed for a mean of 5.7 years, the annual incidence of pleural effusion was found to be 0.34% in the women and 1.54% in the men. In 4 pleural fluids the fluid-to-serum ratio for Waaler-Rose titres was 1:4, which agrees with the ratios for other proteins and does not suggest a pathogenetic role of the rheumatoid factor. In two other pleural fluids the findings suggested immune mechanisms implicating the rheumatoid factor. In general, the data, both clinical and any other, was compatible with the possibility of multiple etiological and pathogenetic factors in rheumatoid pleural effusions. These were: extrinsic factors causing pleural thickenings in 25% of non-rheumatoid men and in less than 10% of non-rheumatoid women aged more than 50 years; preceding lung disorder; particular inflammatory responsiveness due to rheumatoid disease, related or unrelated to rheumatoid factor; and some degree of transudation.

Lysozyme (muramidase) in pleural effusions and serum.

The authors evaluated the usefulness of paired pleural fluid and serum lysozyme determination in the differential diagnosis of pleurisies in 118 patients. Lysozyme activity of tuberculous effusions was found significantly higher (P less than 0.001) than that of effusions due to malignancy or heart failure, but considerable overlap of the individual values was observed. All patients with tuberculous pleurisy or tuberculous empyema, as well as a group of patients with suspected tuberculous effusion, had pleural fluid to serum lysozyme ratio of 1.0 or greater. In the remaining groups, only three patients, one with malignancy, one with lupus erythematosus, and one with sarcoidosis, exceeded this value. Their results suggest that pleural fluid to serum lysozyme ratio can be applied effectively in the differential diagnosis of tuberculous pleurisy.

Adenosine deaminase estimations in the differentiation of pleural effusions.

Adenosine deaminase (ADA) estimations were performed on the pleural fluid from 368 effusions. The mean (+/-SD) ADA concentration in tuberculous effusions was 92,11 +/- 37,05 U/l, and these values were found to be highly statistically different from the 23,23 +/- 13,15 U/l in secondary malignant tumours of the pleura, the 34,86 +/- 14,2 U/l in mesotheliomas, and the 23,81 +/- 15,07 U/l in pulmonary embolism. The ADA values of 64,3 +/- 44,95 U/l in lymphoproliferative disorders were less significantly different. No statistical difference could be found between values in the tuberculous group and the ADA levels of 97,57 +/- 82 U/l found in para-infective effusions, but these could be distinguished from each other by microscopic examination of the pleural fluid. The importance of ADA estimations in the diagnosis and differentiation of tuberculous effusions is discussed and the role of microscopy is emphasized.