Pulmonary hemorrhage. An uncommon cause of pulmonary infiltrates in patients with AIDS.

We describe two patients with AIDS who developed new diffuse pulmonary infiltrates during the course of their hospitalization. In both cases, the infiltrates were attributed to pulmonary hemorrhage complicating an existing condition rather than representing a new pulmonary process. Identification of pulmonary hemorrhage in these patients allowed for discontinuation of treatment with empiric medications and continued appropriate supportive care.

Oncogenes and growth factors in human lung cancer.

We are only beginning to understand the importance of lung cancer tumor biology in relation to prognosis and response to therapy. Many of the biologic and genetic changes we have described are preliminary observations and require further confirmation before clinical use. However, information concerning three oncogenes may soon prove to be helpful in the clinical arena: the myc genes in SCLC, and the ras genes and c-erbB-2 in NSCLC. In general their presence identifies poor patient response to therapy and poor survival. These markers are currently being used in a clinical setting at some research centers, but are not recommended for general diagnostic or prognostic use without further confirmation of their utility. Incorporation of this information with that learned by standard staging procedures may result in improved understanding of patient prognosis and challenge current concepts of lung cancer treatment. For example, surgically resected stage I NSCLC patients may benefit from adjuvant therapy if found to have these adverse biologic factors, and require more stringent follow-up after therapy. Finally the understanding of the pathogenesis of lung cancer may enable the development of novel therapy directed against these growth pathways. Our ultimate goal is to derive a therapeutic and prognostic paradigm involving both molecular-genetic and clinical factors to arrive at an optimal staging model and treatment plan.

Prognostic significance of tumor proliferative fraction and DNA content in stage I non-small cell lung cancer.

Analyses of tumor DNA content and proliferative fraction by flow cytometry have been useful as prognostic determinants in a variety of solid tumors. The significance of this analysis in Stage I (T1N0M0 and T2N0M0) non-small cell lung carcinoma (NSCC) is unestablished. We determined DNA content (ploidy) and proliferative fraction (percentage S phase) on 44 surgically resected Stage I NSCC specimens obtained between 1977 and 1982. All cases had a minimum follow-up of 5 yr. Of the 44 cases, 27 were adenocarcinomas, 15 squamous cell carcinomas, and 2 large cell carcinomas. Of these, 32 (73%) had T1N0M0 lesions and 12 (27%) had T2N0M0 lesions. Overall 5-yr survival was 70%. All patients surviving 5 yr were free of detectable tumor. Patients with T1N0M0 lesions had an 81% 5-yr survival, but those with T2N0M0 lesions had a 42% 5-yr survival (p = 0.009). Analysis of tumor DNA content revealed 35 diploid tumors (79%) and 9 aneuploid tumors (21%). The 5-yr survival for diploid tumors was 77% compared with a 44% 5-yr survival in aneuploid lesions (p = 0.048). The median proliferative fraction was 6%. All patients with a percentage S phase less than 6% survived 5 yr, and 41% (9 of 22) of those greater than 6% survived 5 yr (p less than 0.001). When 8% S phase was used as a cutoff, 93% (28 of 30) below the cutoff survived 5 yr but only 21% (3 of 14) above the cutoff survived 5 yr (p less than 0.001).(ABSTRACT TRUNCATED AT 250 WORDS)

Macrophage colony-stimulating factor (CSF-1) enhances invasiveness in CSF-1 receptor-positive carcinoma cell lines.

We have identified two lung carcinoma cell lines, A549 and Calu-1, expressing low levels of the macrophage colony-stimulating factor (CSF-1) receptor (CSF-1R), encoded by the c-fms oncogene. The effect of CSF-1 on the invasive potential of these CSF-1R-positive tumor cell lines and on two other CSF-1R-bearing cell lines, the BT-20 breast carcinoma cell line and the CSF-1 growth-dependent murine macrophage cell line BAC1.2F5, was examined using a human amnionic basement membrane invasion model. Culture of A549, Calu-1, and BAC1.2F5 cells with CSF-1 (250 ng/ml) resulted in a maximal 12-, 5-, and 12-fold enhancement of invasion, respectively, compared to control cells cultured in medium alone. Larger concentrations of CSF-1 (750 ng/ml) reduced A549 and Calu-1 invasiveness compared to the effect of the 250-ng/ml dose. Maximal enhancement in invasion of A549 and Calu-1 cells occurred after a 24- and 48-h exposure to CSF-1, respectively. CSF-1 increased invasiveness 6-fold in BT-20 cells induced by glucocorticoids to express high levels of CSF-1R, in comparison to control cells not exposed to glucocorticoids or CSF-1. In contrast, CSF-1 had no effect on invasion in the CSF-1R-negative MCF-7 cell line. Culture of A549 and Calu-1 cells with other cytokines and growth factors including GM-CSF (500 units/ml), IL-3 (1 ng/ml), interferon-gamma (500 units/ml), and tumor necrosis factor (50 units/ml) had no significant effect on invasiveness. Thus, CSF-1 increases invasiveness in CSF-1R-positive tumor cell lines, suggesting a role in enhancing the metastatic potential of tumor cells expressing the CSF-1R.

Endogenous receptor-bound urokinase mediates tissue invasion of the human lung carcinoma cell lines A549 and Calu-1.

Macrophage colony-stimulating factor (CSF-1) increases the tissue invasive potential of the CSF-1 receptor-bearing lung carcinoma cell lines A549 and Calu-1 by increasing the number of endogenously bound urokinase-type plasminogen activators (u-PA)s on these cells. CSF-1, at concentrations which optimize invasion of A549 and Calu-1 cells into human amnion membranes (250 ng/ml), maximally augments the number of u-PA receptors occupied by endogenously produced urokinase. Preincubation of A549 and Calu-1 cells with the anti-u-PA monoclonal antibody MPW5UK (25 micrograms/ml) or with a 20- to 40-fold stoichiometric excess of fluid phase type 2 plasminogen activator inhibitor abrogates invasiveness, indicating that functionally active cell surface u-PA is essential for tissue invasion. In contrast, fluid phase type 1 plasminogen activator inhibitor (PAI-1, 5-15 units/ml) does not inhibit invasiveness unless preincubated with the amnion membranes. Inhibition of invasion by PAI-1 is abolished by presaturating the amnion membranes with antiitronectin monoclonal antibody (10 micrograms/ml) which prevents binding of PAI-1 to tissue-associated vitronectin. Binding of PAI-1 to tissue vitronectin is therefore a prerequisite for its inhibitory action. Thus, endogenously receptor-bound u-PA is the primary protease mediating CSF-1-induced tissue invasiveness of the lung carcinoma cell lines A549 and Calu-1.

Characterization of colony stimulating factor activity in the human respiratory tract. Comparison of healthy smokers and nonsmokers.

The number and function of pulmonary macrophages are critical to lung homeostasis. To characterize factors normally present in the human respiratory tract that can influence these parameters, bronchoalveolar lavage (BAL) fluid obtained from healthy smokers and nonsmokers was assayed for the presence of colony-stimulating factor (CSF) activity. Concentrated BAL fluid from both populations was capable of inducing incorporation of [3H]thymidine by murine macrophages. The mean increase (+/- SEM) in incorporation over control cultures not exposed to BAL fluid was 0.98 +/- 0.22 for nonsmokers and 2.25 +/- 1.19 for smokers (p less than 0.001). This CSF bioactivity was characterized as macrophage-CSF (M-CSF) by virtue of its action on murine macrophages, the detection of M-CSF protein by a specific ELISA assay, and the inability to detect other macrophage-active CSFs, granulocyte macrophage-CSF (GM-CSF) and interleukin-3 (IL-3), in a proliferation assay employing the MO7E cell line. There was a significant correlation between macrophage number in BAL samples and measureable bioactivity among both smokers and nonsmokers (r = 0.763; p less than 0.001). This suggested that macrophages themselves are a source of the M-CSF detected in BAL fluid. To examine this possibility, slot-blot analysis of macrophage RNA was performed. Constitutive expression of comparable amounts of M-CSF mRNA and protein was found in cells from both smokers and nonsmokers. However, macrophages obtained from a randomly selected subset of four smokers but none of five nonsmokers exhibited increased production of M-CSF in response to an inflammatory stimulus, lipopolysaccharide (LPS; 5 ng/ml). M-CSF added to macrophage cultures was degraded by nonsmokers' cells as expected over 24 h.(ABSTRACT TRUNCATED AT 250 WORDS)

Colony-stimulating factor induction of protooncogene expression in rat alveolar macrophages.

The effect of granulocyte-macrophage colony-stimulating factor (GM-CSF) and macrophage colony-stimulating factor (M-CSF) on the expression of c-fos and c-myc protooncogenes was studied in rat alveolar macrophages (AM). AM were exposed in vitro to GM-CSF (100 U/ml) or M-CSF (1,000 U/ml) for 30-120 min, and c-fos and c-myc mRNA expression was determined by in situ hybridization and Northern blot analysis. GM-CSF caused a rapid induction of c-fos mRNA after 30 min and c-myc mRNA after 60 min. Exposure to M-CSF stimulated maximal expression of c-fos mRNA after 60 min and c-myc mRNA after 120 min. Under the same experimental conditions lipopolysaccharide (100 ng/ml) induced a comparable amount of c-fos and c-myc mRNA expression, whereas culture of AM with medium alone did not induce c-fos or c-myc expression. Thus GM-CSF and M-CSF induce AM in vitro to express the nuclear protooncogenes c-fos and c-myc. This effect of colony-stimulating factors on protooncogene expression may be of importance in the local regulation of AM activation and/or proliferation in an inflammatory lung response.

Malignancies metastatic to the pleura.

In patients over 50 years of age, neoplasms of the pleura are probably the second most common cause of a pleural effusion after congestive heart failure. Lung cancer, breast cancer, lymphoma, ovarian carcinoma, and stomach cancer are the leading causes of malignant pleural disease, and adenocarcinoma is the most common cell type. This review discusses in detail the etiology and incidence, pathogenesis, clinical manifestations, diagnosis, prognosis, and treatment of neoplasms that involve the pleura with special reference to malignant and paramalignant pleural effusions.

Pulmonary and pleural manifestations of extrathoracic malignancies.

Metastatic spread of nonpulmonary malignancies to the lung is a common clinical problem. However, the evaluation and treatment of metastatic lung disease may be confusing. This article considers both pulmonary and pleural metastases. The basic biology of the metastatic process is discussed, together with the pathologic, clinical, radiologic, diagnostic, and therapeutic features of pulmonary metastases. Metastatic disease to the pleura is reviewed in detail, including etiology and incidence, pathogenesis, clinical manifestations, diagnosis, prognosis, and treatment.

ELISA analysis of BACTEC bottles for the earlier diagnosis of tuberculosis.

Enzyme-linked immunosorbent assays (ELISA) have enabled earlier identification of Mycobacterium tuberculosis (TB) in clinical settings by utilizing both TB antibody and antigen detection. We studied the sensitivity and specificity of ELISA detection of TB antigen by using a commercially available anti-BCG antibody in conjunction with BACTEC 7H12B culture bottles. We compared these results against those obtained with cultures of Mycobacterium avium-intracellulare and Mycobacterium kansasii. All BACTEC bottles were inoculated with known concentrations of organisms. TB antigen was detected by ELISA in BACTEC culture bottles of TB 12 days before the BACTEC system could itself identify the species. A growth index of greater than or equal to 10 reliably indicated that tuberculosis antigen was detectable by ELISA. The correlation between growth index and antigen concentration was extremely high (r = 0.95 and p less than 0.001). There was insignificant cross-reactivity with the other atypical mycobacteria at the levels of growth tested. This technique potentially offers a sensitive and specific means for the early identification of TB in BACTEC culture bottles.

Antitumor and toxic effects of combination chemotherapy with bleomycin and a phenothiazine anticalmodulin agent.

Phenothiazines and structurally related calmodulin antagonists acted synergistically with bleomycin in killing malignant cells in culture. For exploration of the potential clinical importance of this observation, the effects of administering the combination of chlorpromazine and bleomycin in vitro and in vivo against the transplantable B16 melanoma were studied. The toxicity of the combination to bone marrow and lungs was also determined. Combined chlorpromazine and bleomycin therapy reduced the concentration of chlorpromazine required to inhibit cellular growth of B16 melanoma cells by greater than half (25 vs. 60 microM). Against the transplanted tumor, after 3 weeks of treatment the tumor sizes (in cubic millimeters) were 540 +/- 100 (vehicle), 520 +/- 120 (chlorpromazine), 170 +/- 10 (bleomycin), and 80 +/- 20 (bleomycin + chlorpromazine, P less than .01 vs. bleomycin alone). The median time required to reach a tumor size of 100 mm3 was 15 days for vehicle and chlorpromazine, 17 days for bleomycin, and 26 days for chlorpromazine and bleomycin together. The doubling time of the tumor was also increased 3.5-fold above control with the two-drug combination. At doses of bleomycin and chlorpromazine that increased antitumor activity, no pulmonary fibrosis, measured histologically and biochemically, was detected. At higher doses of bleomycin, the addition of chlorpromazine diminished lung fibrosis. The combination of drugs was not more toxic to hematopoietic precursors than chlorpromazine alone. These studies suggested that the addition of a phenothiazine to bleomycin can augment the therapeutic efficacy of bleomycin in vivo without substantially increasing its toxicity.

Alterations in pulmonary protective enzymes following systemic bleomycin treatment in mice.

Repeated bleomycin administration in animals and humans produces significant lung fibrosis. The pathogenesis of this toxicity may be multifactorial, but it appears to be initiated through the production of radical oxygen species by an activated bleomycin-iron-oxygen ternary complex. Protection of lung tissue from bleomycin-induced toxicity may occur through both specific metabolic inactivation of bleomycin by the enzyme bleomycin hydrolase, as well as by such non-specific antioxidants as catalase and the glutathione system. The effect of chronic, systemic administration of bleomycin on the activities and levels of these enzymes and proteins in pulmonary tissue is unknown. C57BL/6 mice were injected subcutaneously with saline, non-fibrogenic (2 mg/kg) and fibrogenic (10 mg/kg) doses of bleomycin twice-weekly for 6 weeks. Animals were killed at 0, 1.5, 3, and 6 weeks after initiation of bleomycin treatment. Catalase activity was increased more than 50% at 3 weeks in the low-dose animals, and was decreased over 40% at 6 weeks in the high-dose animals. Total lung glutathione levels were unaffected in both groups, although glutathione reductase activity was increased significantly (over 2-fold) at 1.5 and 3 weeks in the high-dose animals. At 6 weeks glutathione reductase was increased 7- and 12-fold in low and high-dose animals respectively. Glutathione peroxidase activity also was elevated more than 2-fold above control values at 6 weeks in both sets of animals. There was no evidence of induction of bleomycin hydrolase activity at any time point. Rather, bleomycin hydrolase activity was decreased significantly to 30 and 40% of control values at 3 and 6 weeks, respectively, in mice receiving the fibrogenic doses of bleomycin. These results demonstrate that chronic, systemic administration of non-fibrogenic and fibrogenic doses of bleomycin produces changes in activity of lung antioxidant defense mechanisms. The early loss of lung bleomycin hydrolase activity may contribute to the pathogenesis of bleomycin-induced pulmonary toxicity following repeated drug exposure.

Lung cancer in the elderly.

Lung cancer is the most common malignancy in the elderly population and was responsible for greater than 125,000 deaths in 1985. Although advances have been made in the areas of diagnosis and staging, the long-term outlook for the disease remains poor, primarily because of dissemination of disease prior to diagnosis. Despite this, several studies suggest that lung cancer in the elderly is a more indolent disease process, which should be treated in the same manner as it is in the young population.

Effect of Fluosol-DA/O2 on the antitumor activity and pulmonary toxicity of bleomycin.

The effect of an oxygen-carrying perfluorochemical emulsion on bleomycin antitumor activity and pulmonary toxicity was examined. Fluosol-DA (0.3 ml/mouse, i.v.), combined with bleomycin (10 or 15 mg/kg, i.p.) and a 2 h exposure to 95% oxygen (BFO) increased by five- to six-fold the tumor growth delay of FSaIIC fibrosarcoma compared to bleomycin alone (B). Only a slight increase in tumor growth delay was noted with the incomplete combinations of bleomycin and O2 (BO) and bleomycin and Fluosol-DA (BF). When bleomycin (10 mg/kg) was co-administered with 0.3 ml Fluosol-DA and 95% O2, cell survival was reduced ten-fold compared to that seen with bleomycin alone. In contrast, the surviving fraction of cells obtained from FSaIIC tumors treated in vivo indicated that 0.3 ml Fluosol-DA per mouse or a 2 h exposure to 95% O2 did not markedly alter the effects of bleomycin alone. The pulmonary effects of the BFO combination were assessed during the course of the therapy by bronchoalveolar lavage (BAL) analysis and pulmonary hydroxyproline (OH-Pro) content. Mice treated with this combination had a 20-fold increase in total numbers of cells obtained in the BAL compared to control animals. An increased cellularity in the lungs was also seen morphologically. The composition of the cells in the lavage fluid was altered after BFO but not BO treatment and reflected a neutrophilic influx. Furthermore, total protein recovered in the BAL fluid was increased 5-fold in the BFO treatment group compared to that in the control mice. Pulmonary OH-Pro, an index of collagen and fibrosis, was unaffected acutely after three treatments of either BFO or BO compared to control mice. Thus, Fluosol-DA and O2 can enhance the antitumor activity of bleomycin. The increased pulmonary cellularity suggests that this combination may also have adverse effects on lung tissue.