Abnormalities of esterase and glycogen in developing macrophages in non-Hodgkin's lymphoma: a quantitative cytochemical study.

Integrating microdensitometry has been used to quantitate changes in 4 cytoplasmic enzymes (NADH dehydrogenase, succinate dehydrogenase, acid phosphatase and alpha-naphthyl butyrate esterase), DNA, RNA and glycogen in developing macrophages from 17 patients with non-Hodgkin's lymphoma and 19 normal subjects. Cytochemical measurements were made at intervals over 6 days of suspension culture; over 16 000 individual cells were examined in total and the results subjected to analysis of variance. While the levels of enzymes and RNA of both groups showed increases over the period of culture, the levels of alpha-naphthyl butyrate esterase in the patients' cells were consistently lower than the corresponding values for the normal cells and glycogen levels were higher, these differences satisfying the pre-determined requirements for statistical significance. It is concluded that (a) maturational changes take place in cytochemical constituents of developing macrophages of non-Hodgkin's lymphoma (b) there are disturbances affecting the amounts of the specific enzyme alpha-naphthyl butyrate esterase and glycogen (c) these abnormalities may be part of a compromise of host defense mechanisms by the disease, although a pre-existing defect in esterase increasing the susceptibility to malignancy is another possibility, and (d) the methods used may be of value in future investigations of the cause of the disturbances and their correction.

Quantitative enzyme cytochemistry during human macrophage development.

Integrating microdensitometry was used to study changes in the intracellular activity of 4 enzymes during macrophage development. Suspension cultures of blood monocytes from 19 healthy human subjects were examined at 0, 2, 4 and 6 d. Mononuclear phagocytes were harvested by glass adherence and standard methods were used for cytochemical staining for NADH dehydrogenase, succinate dehydrogenase, acid phosphatase and alpha-naphthyl butyrate esterase. All specimens from all subjects were stained at the same time and staining intensities in individual cells were measured at appropriate wavelengths. A highly significant increase in enzyme activity with culture time was found for all 4 enzymes. These increases in mitochondrial, lysosomal and ectoenzyme activities during development indicate the increasing functional capabilities of the macrophages.

Disordered macrophage development in Hodgkin's disease shown by quantitative cytochemistry.

In order to investigate the disordered maturation of mononuclear phagocytes previously found in Hodgkin's disease, integrating microdensitometry was used to quantitate changes in seven cytochemical constituents (NADH dehydrogenase, succinate dehydrogenase, acid phosphatase, alpha-naphthyl butyrate esterase, DNA, RNA and glycogen) of developing macrophages from 19 patients and 19 normal subjects. Individual cells were studied at intervals over six days of suspension culture; the results were subjected to analysis of variance. In both groups, all the constituents studied except DNA showed highly significant increases over the period of culture. Consistently lower levels of alpha-naphthyl butyrate esterase (approximately 65%) and increased levels of glycogen were present in the Hodgkin's group. The results show that (1) maturational changes occur in the cytochemical constituents of developing macrophages of Hodgkin's disease, and (2) there are disturbances affecting the specific enzyme alpha-naphthyl butyrate esterase and glycogen that are likely to have profound implications for host defense mechanisms.

Quantitation of immune-mediated erythrophagocytosis and rosetting in developing macrophages.

As part of our ongoing investigations of mononuclear phagocytes in health and disease, measurement of antibody-dependent erythrophagocytosis and rosette formation was carried out during human macrophage development in a suspension culture system. Cells derived from 12 normal human subjects were studied at the monocyte (day 0) and macrophage (day 6) stage of development. Analysis of variance demonstrated significant differences between monocytes and macrophages: the mean proportions of cells (+/- SEM) showing erythrophagocytosis were 0.09 (+/- 0.02) and 0.24 (+/- 0.06), respectively, and the average numbers of engulfed red cells (+/- SEM) were 1.12 (+/- 0.02) and 1.38 (+/- 0.11), respectively. No significant difference was detected in rosette formation. It is concluded that (1) immune-mediated activity of developing macrophages can be quantitated; (2) functional changes take place over 6 days of culture; (3) macrophages show more erythrophagocytic activity than monocytes, and (4) the method should be of value in making quantitative comparisons of their functional development in haematological and other disease states.

Increase in RNA and glycogen but not in DNA in developing human macrophages proven by integrating microdensitometry.

In order to quantitate DNA, RNA and glycogen contents of developing macrophages, blood monocytes were obtained from 19 healthy human subjects and examined after 0, 2, 4 and 6 days of suspension culture. Cytochemical staining was carried out by standard methods using Feulgen, cuprolinic blue and PAS. All specimens from all subjects were stained at the same time. Examination was carried out in an integrating microdensitometer, the staining intensities of individual cells being measured at appropriate wavelengths. Over the 6 days of culture, highly significant increases took place in RNA and glycogen, but no significant change was found in DNA content. These findings are taken to indicate that increased protein synthesis and the building up of fuel reserves are features of macrophage development but that S phase DNA replication does not occur.

Immune-mediated interactions during macrophage development in non-Hodgkin's lymphoma.

As part of an investigation of mononuclear phagocytes in malignant lymphoma, measurement of immune-mediated erythrophagocytosis and rosette formation was carried out on cells grown in suspension culture at the monocyte (Day 0) and macrophage (Day 6) stages; the culture medium contained autologous serum. Cells were derived from 10 patients with untreated non-Hodgkin's lymphoma (NHL) and from 12 normal individuals. The results were subjected to Analysis of Variance and demonstrated a significant difference between the two groups with respect to erythrophagocytosis but not to rosette formation. In the NHL group, the proportion of erythrophagocytic cells showed no significant increase between the monocyte and macrophage stages (0.07 to 0.09), in contrast to the marked increase seen in the normal group (0.09 to 0.24). In a pilot investigation to examine the possible role of factors in the serum, cells derived from the NHL patients were cultured with serum from healthy donors; they showed no significant difference in the immune-mediated functions from those grown in autologous serum. Overall, the results provide further quantitative evidence of defective macrophage maturation in NHL, presumably reflecting the compromise of host defence mechanisms.

Bone tuberculosis: results and experience in Leicestershire.

Management of tuberculosis falls to the thoracic physician and includes extrapulmonary sites. We have conducted a survey of notification of bone tuberculosis in Leicestershire from 1978 to 1983 inclusive. Eighty-one cases were notified, 75 were traced and 69 confirmed tuberculosis (mean age 40.2 years, 39 male, 54 from the Indian sub-continent (ISC)). Mean length of symptoms was 8.4 months, the commonest being pain and swelling. The diagnosis was established by evidence of bone involvement plus at least one of the following: positive Heaf test (54/56), positive culture at bone sites for Mycobacterium tuberculosis (45/47), or suggestive histology (30/34). There were no resistant organisms. Chemotherapy (mean duration 16.2 months) was well tolerated. Spine was the site most often involved (37 patients, 34 ISC), with abscess formation common (17 patients). Four who presented with spinal cord compression received a mean of 8 months' bed rest, three required decompression and one died. No patient developed new neurological signs on treatment. Fourteen others received an average of 2.3 months' bed rest. In this predominantly ISC immigrant population, spinal tuberculosis is common but does not progress to cord compression on treatment. For those without cord compression the course and outcome were similar whether or not they were treated with bed rest.

Hepatic necrosis with doxapram hydrochloride.

We report the first case of acute hepatic necrosis, which we believe to have been caused by the administration of the respiratory stimulant, doxapram hydrochloride (Dopram).

Thoracic lymphadenopathy in Asians resident in the United Kingdom: role of mediastinoscopy in initial diagnosis.

Although the incidence of superficial glandular tuberculosis is high in Asian immigrants, a clinical diagnosis without biopsy has previously been shown to be inaccurate in 22% of cases. The role of diagnostic mediastinoscopy and biopsy in thoracic lymphadenopathy was therefore evaluated in 41 consecutive Asian patients. Tuberculosis was diagnosed by histological examination or culture (or both) of gland biopsy material in 24 (59%). A further 12 patients, however, also received antituberculous chemotherapy, with a response in 10 cases. If these are included, 34 (83%) were finally considered to have tuberculosis. The symptoms and ages of the patients with tuberculosis were similar to those typically seen in caucasians with sarcoidosis. Six had tuberculous bilateral hilar lymphadenopathy. In only four cases (10%) was a positive diagnosis other than tuberculosis established; in two (lymphoma and thymoma) mediastinoscopy confirmed preoperative suspicions, and saved only two more (with sarcoidosis and vascular anomaly) from unnecessary antituberculous treatment. Complications included severe haemorrhage (1) and chronic tuberculous sinus in the endoscopy tract (2). Mediastinoscopy is unlikely to change management in most patients, produces an appreciable amount of morbidity, and should be reserved for cases in which there is additional clinical doubt. Any future decline in the prevalence of tuberculosis in Asians may, however, require its further evaluation.

Gastrointestinal tuberculosis.

Gastrointestinal tuberculosis continues to occur sporadically in hospitals in the British Isles. Accurate diagnosis may enable its rare but lethal complications to be anticipated. Pathology, clinical features and treatment are reviewed. The use of antituberculous chemotherapy and corticosteroids is discussed with reference to 14 cases of gastrointestinal tuberculosis seen in Southampton between 1951 and 1973.