Combination of CT and telomerase+ circulating tumor cells improves diagnosis of small pulmonary nodules.

BACKGROUNDEarly diagnosis and treatment are key to the long-term survival of lung cancer patients. Although CT has significantly contributed to the early diagnosis of lung cancer, there are still consequences of excessive or delayed treatment. By improving the sensitivity and specificity of circulating tumor cell (CTC) detection, a solution was proposed for differentiating benign from malignant pulmonary nodules.METHODSIn this study, we used telomerase reverse transcriptase-based (TERT-based) CTC detection (TBCD) to distinguish benign from malignant pulmonary nodules < 2 cm and compared this method with the pathological diagnosis as the gold standard. FlowSight and FISH were used to confirm the CTCs detected by TBCD.RESULTSOur results suggest that CTCs based on TBCD can be used as independent biomarkers to distinguish benign from malignant nodules and are significantly superior to serum tumor markers. When the detection threshold was 1, the detection sensitivity and specificity of CTC diagnosis were 0.854 and 0.839, respectively. For pulmonary nodules ≤ 1 cm and 1-2 cm, the sensitivity and specificity of CTCs were both higher than 77%. Additionally, the diagnostic ability of CTC-assisted CT was compared by CT detection. The results show that CT combined with CTCs could significantly improve the differentiation ability of benign and malignant nodules in lung nodules < 2 cm and that the sensitivity and specificity could reach 0.899 and 0.839, respectively.CONCLUSIONTBCD can effectively diagnose pulmonary nodules and be used as an effective auxiliary diagnostic scheme for CT diagnosis.FUNDINGNational Key Research and Development Project grant nos. 2019YFC1315700 and 2017YFC1308702, CAMS Initiative for Innovative Medicine grant no. 2017-I2M-1-005, and National Natural Science Foundation of China grant no. 81472013.

A Novel In Vitro Mouse Model to Study Mycobacterium tuberculosis Dissemination Across Brain Vessels: A Combination Granuloma and Blood-Brain Barrier Mouse Model.

In vitro culture models of the blood-brain barrier (BBB) provide a useful platform to test the mechanisms of cellular infiltration and pathogen dissemination into the central nervous system (CNS). We present an in vitro mouse model of the BBB to test Mycobacterium tuberculosis (Mtb) dissemination across brain endothelial cells. One-third of the global population is infected with Mtb, and in 1%-2% of cases bacteria invade the CNS through a largely unknown process. The "Trojan horse" theory supports the role of a cellular carrier that engulfs bacteria and carries them to the brain without being recognized. We present for the first time a protocol for an in vitro BBB-granuloma model that supports the Trojan horse mechanism of Mtb dissemination into the CNS. Handling of bacterial cultures, in vivo and in vitro infections, isolation of primary astroglial and endothelial cells, and assembly of the in vitro BBB model is presented. These techniques can be used to analyze the interaction of adaptive and innate immune system cells with brain endothelial cells, cellular transmigration, BBB morphological and functional changes, and methods of bacterial dissemination. © 2020 Wiley Periodicals LLC. Basic Protocol 1: Isolation of primary mouse brain astrocytes and endothelial cells Basic Protocol 2: Isolation of primary mouse bone marrow-derived dendritic cells Support Protocol 1: Validation of dendritic cell purity by flow cytometry Basic Protocol 3: Isolation of primary mouse peripheral blood mononuclear cells Support Protocol 2: Isolation of primary mouse spleen cells Support Protocol 3: Purification and validation of CD4+ T cells from PBMCs and spleen cells Basic Protocol 4: Isolation of liver granuloma supernatant and determination of organ load Support Protocol 4: In vivo and in vitro infection with mycobacteria Basic Protocol 5: Assembly of the BBB co-culture model Basic Protocol 6: Assembly of the combined in vitro granuloma and BBB model.

Use of TBAg/PHA ratio in distinguishing tuberculoma from cancer in solitary pulmonary nodule or mass.

INTRODUCTION: Differentiation of tuberculoma from cancer in solitary pulmonary nodule or mass still remains a major challenge in diagnostic laboratories. OBJECTIVES: The objective of this study is to determine the performance of T-SPOT.TB assay in discriminating these 2 diseases. METHODS: We prospectively enrolled 331 patients with a solitary pulmonary nodule or mass on computed tomography scans. Conventional tests and T-SPOT.TB assay were simultaneously performed in all participants. RESULTS: Our results showed that the performance of directly using T-SPOT.TB results in distinguishing tuberculoma from cancer in solitary pulmonary nodule or mass was not satisfactory because of moderate sensitivity and specificity. However, a further calculation of the ratio of TB-specific antigen (TBAg) to phytohemagglutinin (PHA) (TBAg/PHA ratio) of T-SPOT.TB assay may lead to improvement in distinguishing these 2 diseases. If using the threshold value of 0.236, the sensitivity and specificity of the TBAg/PHA ratio in distinguishing tuberculoma from cancer in solitary pulmonary nodule or mass were, respectively, 80.6% and 93.3%. The area under the curve (AUC) of the receiver operating characteristic curve was 0.921 (95% confidence interval, 0.875-0.967). Furthermore, the TBAg/PHA ratio may also be used to distinguish tuberculoma from other benign diseases (AUC: 0.909, sensitivity: 85.07%, specificity: 90%). CONCLUSIONS: Calculation of the TBAg/PHA ratio might provide a useful non-invasive tool for distinguishing tuberculoma from cancer in patients with a solitary pulmonary nodule or mass in TB-endemic countries.

Modeling nanoparticle delivery of TB drugs to granulomas.

Tuberculosis, which typically presents as a pulmonary disease, has a complex pathology. The primary site of infection, the Ghon focus, recruits immune cells and a granuloma forms. At earlier stages the granuloma is still vascularized, offering the best opportunity for drug treatment. In the more progressive state blood flow is reduced and a distinct caseous structure develops. Effective delivery of drugs to bacilli in the core of the granuloma becomes very difficult. It is perceivable that granuloma cores could create conditions where bacilli persist and develop resistance. In this study we analyze drug delivery to granulomas by means of a nanoparticle delivery system. The model consists of two parts; the overall distribution of the nanoparticles is described by a simple circulatory model and this result is used in the second part, focusing on transport in a capillary lined with macrophages. Nanoparticles enter the macrophages where they are metabolized and the drugs are released. The model reveals significant differences in drug concentrations between the plasma and macrophages. Based on the results of the model, strategies for improved drug delivery are proposed.

Metabolomic signatures in guinea pigs infected with epidemic-associated W-Beijing strains of Mycobacterium tuberculosis.

With the understanding that the laboratory propagated strain of Mycobacterium tuberculosis H37Rv is of modest virulence and is drug susceptible, in the present study, we performed a nuclear magnetic resonance-based metabolomic analysis of lung tissues and serum obtained from guinea pigs infected by low dose aerosol exposure to clinical isolates of Mycobacterium tuberculosis. High Resolution Magic Angle Spinning NMR coupled with multivariate statistical analysis of 159 lung tissues obtained from multiple locations of age-matched naïve and 30 and 60 days of infected guinea pig lungs revealed a wide dispersal of metabolic patterns, but within these, distinct clusters of signatures could be seen that differentiated between naive control and infected animals. Several metabolites were identified that changed in concert with the progression of each infection. Major metabolites that could be interpreted as indicating host glutaminolysis were consistent with activated host immune cells encountering increasingly hypoxic conditions in the necrotic lung lesions. Moreover, glutathione levels were constantly elevated, probably in response to oxygen radical production in these lesions. Additional distinct signatures were also seen in infected serum, with altered levels of several metabolites. Multivariate statistical analysis clearly differentiated the infected from the uninfected sera; in addition, Receiver Operator Characteristic curve generated with principal component 1 scores showed an area under the curve of 0.908. These data raise optimism that discrete metabolomic signatures can be defined that can predict the progression of the tuberculosis disease process, and form the basis of an innovative and rapid diagnostic process.

[Dynamic contrast-enhanced CT patterns and pathologies of noncalcified pulmonary tuberculomas].

OBJECTIVE: To analyze characteristic CT enhancement patterns of noncalcified pulmonary tuberculomas and their pathological basis. METHOD: Fifty-six patients with noncalcified pulmonary tuberculomas underwent surgical resection of the tuberculomas. Enhanced CT images of these tuberculomas were reviewed and analyzed in relation to the histological findings. RESULTS: Of the 56 patients, 45 showed no enhancement in the tuberculomas, which were histologically characterized by central caseous necrosis and a poorly vascularized peripheral fibrotic zone. Eleven patients showed ring-like or eggshell enhancement, and the central low density region was histologically confirmed to be caused by caseous or liquefied necrosis, while the ring enhancement resulted pathologically from moderately or well vascularized peripheral fibrotic or granulomatous tissues. CONCLUSIONS: Pulmonary tuberculomas consists mainly of caseous necrotic tissues characterized by no enhancement and ring or eggshell enhancement on dynamic contrast-enhanced CT.

DTI derived indices correlate with immunohistochemistry obtained matrix metalloproteinase (MMP-9) expression in cellular fraction of brain tuberculoma.

We performed in vivo diffusion tensor imaging (DTI) in a total of 33 patients with brain tuberculomas (BT). Thirteen of them had surgical excision of the lesion as it was clinically indicated, and in these samples matrix metalloproteinase-9 (MMP-9) expression was quantified. We correlated the results of DTI indices like fractional anisotropy (FA), mean diffusivity (MD), linear anisotropy (CL), planar anisotropy (CP) and spherical anisotropy (CS) with MMP-9 expression. In addition, the remaining 20 patients had serial DTI studies while on specific anti-tuberculous drug therapy and DTI indices in these patients were quantified. The FA, CL and CP significantly decreased while MD and CS significantly increased in BT compared to normal white matter. The FA, CL and CP showed negative correlation with MMP-9 while CS correlated positively. In serial follow-up studies in 20 patients FA, CL and CP showed significant increase while CS decreased significantly over time. We conclude that DTI indices show strong correlation with MMP-9 and these may be used as a surrogate marker of MMP-9 expression in BT. In addition, these indices may be of value in assessing the therapeutic response in patients with BT who are treated only with specific anti-tuberculous drugs.

Monocyte-dependent oncostatin M and TNF-alpha synergize to stimulate unopposed matrix metalloproteinase-1/3 secretion from human lung fibroblasts in tuberculosis.

Leukocyte-derived matrix metalloproteinases (MMP) are implicated in the tissue destruction characteristic of tuberculosis (TB). The contribution of lung stromal cells to MMP activity in TB is unknown. Oncostatin M (OSM) is an important stimulus to extrapulmonary stromal MMP induction, but its role in regulation of pulmonary MMP secretion or pathophysiology of TB is unknown. We investigated OSM secretion from Mycobacterium tuberculosis (Mtb)-infected human monocytes/macrophages and the networking effects of such OSM on lung fibroblast MMP secretion. Mtb increased monocyte OSM secretion dose dependently in vitro. In vivo tuberculous granulomas immunostained positively for OSM. Further, conditioned media from Mtb-infected monocytes (CoMTb) induced monocyte OSM secretion (670 +/- 55 versus 166 +/- 14 pg/mL in controls), implicating an autocrine loop. Mtb-induced OSM secretion was prostaglandin (PG) sensitive, and required activation of surface G-protein coupled receptors. OSM induction was ERK MAP kinase dependent, p38-requiring but JNK-independent. OSM synergized with TNF-alpha, a key cytokine in TB granuloma formation, to stimulate pulmonary fibroblast MMP-1/-3 secretion, while suppressing secretion of tissue inhibitors of metalloproteinases-1/-2. In summary, Mtb infection of monocytes results in PG-dependent OSM secretion, which synergizes with TNF-alpha to drive functionally unopposed fibroblast MMP-1/-3 secretion, demonstrating a previously unrecognized role for OSM in TB.

Vascular endothelial growth factor as a marker of disease activity in neurotuberculosis.

Vascular endothelial growth factor (VEGF) is a potent angiogenesis mediator. Scant reports are available defining the role of VEGF in active and inactive tubercular meningitis (TBM) with no studies on brain tuberculoma. We quantified VEGF levels by enzyme linked immunoassay (ELISA) in cerebrospinal fluid (CSF) and serum in 20 cases each with active and inactive TBM as well as 22 cases of intraparenchymal tuberculoma. VEGF expression and microvessel angiogenesis quantification was done in 7 cases where tuberculomas were excised. Significantly increased VEGF levels in CSF were found in active TBM cases (106.0+/-50.0 pg/ml) compared to inactive TBM cases (14.7+/-10.0 pg/ml) (p<0.001). Mean serum VEGF levels in active TBM, inactive TBM and tuberculoma were 694.93+/-820.66 pg/ml, 499.61+/-238.33 pg/ml and 541.0+/-389.0 pg/ml, respectively. Immunohistochemical staining of excised tuberculoma demonstrated high expression of VEGF in granulomatous areas with intense positivity in inflammatory mononuclear cells, Langhan's giant cells as well as reactive astrocytes and fibrocytes. A strong positive correlation was observed between microvessel density and VEGF expression. Serial decrease in serum VEGF levels was observed with increasing duration of therapy in tuberculoma. We conclude that increased CSF and serum VEGF levels are a measure of activity of the disease in neurotuberculosis and its gradual decrease over a period of time is probably an indicator of therapeutic response.

Monocyte-dependent fibroblast CXCL8 secretion occurs in tuberculosis and limits survival of mycobacteria within macrophages.

CXCL8 is a chemokine that is implicated in the formation of tuberculous (TB) granulomas and in immunity to Mycobacterium tuberculosis (Mtb). Fibroblast chemokine secretion is important for modulating inflammatory responses in chronic lung disease and inflammatory arthritis but has not been investigated in the pathophysiology of TB. In this study, we used a cellular model to examine monocyte/macrophage-dependent stimulation of fibroblasts by Mtb in the regulation of chemokine secretion, particularly that of CXCL8. Human lung fibroblasts grown in collagen were stimulated with conditioned medium from Mtb-infected monocytes (CoMTb). CoMTb-induced prolonged dose-dependent, p38-mediated expression of stable CXCL8 mRNA by fibroblasts accompanied by a >10-fold increase in CXCL8 secretion (487 +/- 88 ng/ml vs 48.6 +/- 34 ng/ml in controls) at 120 h. Fibroblasts strongly expressed CXCL8 in vivo in human TB granulomas. Inhibition of TNF-alpha or IL-1 in CoMTb abrogated the induction of CXCL8 at a pretranscriptional level. CXCL8 secretion was NF-kappaB, C/EBP, and JNK dependent. Sustained NF-kappaB activation was demonstrated beyond 24 h in response to CoMTb. Exogenous CXCL8 reduced the survival of Mtb within macrophages, and inhibition of CXCL8 was associated with intracellular mycobacterial proliferation. These data show that fibroblasts have a previously unrecognized role in modulating inflammation in TB by their CXCL8-dependent contribution to cell recruitment and mycobacterial killing within the granuloma.

Expression of mycobacterial cell division protein, FtsZ, and dormancy proteins, DevR and Acr, within lung granulomas throughout guinea pig infection.

The ability of Mycobacterium tuberculosis to persist in a dormant state is a hallmark of tuberculosis. An insight into the expression of mycobacterial proteins will contribute to our understanding of bacterial physiology in vivo. To this end, the expression of FtsZ, Acr and DevR was assessed in the lung granulomas of guinea pigs infected with M. tuberculosis. Antigen immunostaining was then compared with the detection of acid-fast bacilli (AFB) and mycobacterial DNA. Surprisingly, immunostaining for all three antigens was observed throughout the course of infection; maximum expression of all antigens was noted at 20 weeks of infection. The intensity of immunostaining correlated well with the presence of intact bacteria, suggesting that mycobacterial antigens in the extracellular fraction have a short half-life; in contrast to protein, extracellular bacterial DNA was found to be more stable. Immunostaining for bacterial division and dormancy markers could not clearly distinguish between replicating and non-replicating organisms during the course of infection. The detection of Acr and DevR from 4 weeks onwards indicates that the dormancy proteins are expressed from early on in infection. Both antigen staining and DNA detection from intact bacilli were useful for detecting intact mycobacteria in the absence of AFB.

[Microvessel density and expressions of survivin and vascular endothelial growth factor in non-small cell lung cancer and their correlations to clinicopathologic features].

BACKGROUND & OBJECTIVE: Survivin, an anti-apoptosis gene, expresses in most tumors, and takes part in tumor angiogenesis. This study was to investigate microvessel density (MVD) and expressions of Survivin and vascular endothelial growth factor (VEGF) in non-small cell lung cancer (NSCLC), and explore their correlations to clinicopathologic features of NSCLC. METHODS: MVD and expressions of Survivin and VEGF in 96 specimens of NSCLC tissues, 31 specimens of tumor adjacent tissues, and 20 specimens of benign lesions were detected by SP immunohistochemistry; their interrelations and correlations to clinicopathologic features of NSCLC were analyzed. RESULTS: Positive rate of Survivin was significantly higher in NSCLC than in adjacent tissues and benign lesions (69.8% vs. 16.1% and 0, P<0.05); its expression was related with differentiation and TNM stage of NSCLC. Positive rate of VEGF was significantly higher in NSCLC than in adjacent tissues and benign lesions (72.9% vs. 45.2% and 25.0%, P<0.05); its expression was related with lymph node metastasis and TNM stage of NSCLC. MVD was significantly higher in NSCLC than in adjacent tissues and benign lesions (24.44+/-7.79 vs. 19.37+/-5.26 and 11.83+/-6.25, P<0.05), and was related with lymph node metastasis and TNM stage of NSCLC. Survivin expression was positively correlated with VEGF expression and MVD. CONCLUSION: Survivin is overexpressed in NSCLC, which relates with differentiation and TNM stage of NSCLC and takes part in angiogenesis.

Relationship of survival, organism containment, and granuloma formation in acute murine tuberculosis.

The relationship among organism growth, immunopathology, and survival was studied in C57BL/6 and A/J mice acutely infected with Mycobacterium tuberculosis (MTB) (Erdman). Although organisms grew at similar rates in the lungs of both mouse strains, A/J mice died prior to 14 days after infection, whereas C57BL/6 mice survived twice as long. The lungs of A/J mice exhibited necrotizing interstitial inflammation and widely distributed acid-fast bacilli without granuloma formation. In contrast, the lungs of C57BL/6 mice had relatively mild interstitial inflammation, which was replaced by focal granulomas, and acid-fast bacilli were primarily within granulomas. MTB induced similar granulomas for A/J and C57BL/6 mice in spleen and liver. In the lung, the A/J mice produced only transient messages for interferon-y (IFN-y), interleukin-1beta (IL-1beta), tumor necrosis factor-alpha (TNF-alpha), IL-10, and inducible nitric oxide synthase (iNOS). The C57BL/6 mice, in contrast, produced a delayed but sustained response in the lung correlating with granuloma onset and characterized by high induction of IL-6, IFN-gamma, IL-1beta, IL-10, and TNF-alpha. Responses in the liver and spleen were also evaluated. These results demonstrate that histopathology and cytokine response to MTB infection varies among organs in mice. Increased survival during acute infection may, therefore, depend on the ability to contain organisms within granulomas in the lung.

Expression of NO-synthase in cells of foreign-body and BCG-induced granulomata in mice: influence of L-NAME on the evolution of the lesion.

The microbicidal activity of macrophages in an inflammatory milieu has been related to the production of a large number of cytokins and intermediary metabolites of oxygen and nitrogen among them, nitric oxide (NO). Considering that granulomatous inflammation is predominantly composed of macrophages and epithelioid cells, we decided to investigate the participation of NO in this peculiar type of inflammation. Two models were used: glass cover slip implantation into the subcutaneous tissue of mice and, the inoculation of live bacillus Calmette-Guérin (BCG) into the footpad of the animals. Using a histochemical method for the detection of NO synthase and of the concentration of citrulin metabolized by cells obtained from cover slips implanted on different time intervals or BCG-activated peritoneal cells, it was possible to demonstrate that epithelioid cells do not produce NO. Cells from granuloma induced by BCG inoculation express NO synthase, with different degrees of reactivity with a higher intensity in the cytoplasm of cells located in the edge of the lesions. The expression of NO synthase in the cytoplasm of these cells decreases with the age of the lesions. It could also be demonstrated that in mice treated with l-name, an inhibitor of NO metabolism, the lesions induced by BCG lost the granulomatous architecture, were necrotic, and had a significant increase in the bacillary load of the lesion. These data allow us to conclude that NO production by macrophages is a determining factor in the organization of the granulomatous lesion and that it also controls the bacterial load in BCG-induced lesions in mice.

[Cellular and subcellular morphology of the repair process in pulmonary tuberculosis]. / Kletochnaia i subkletochnaia morfologiia reparativnykh protsessov pri tuberkuleze legkikh.

The repair processes in pulmonary tissue and granuloma in experimental tuberculosis were studied electron-microscopically and histochemically. At the beginning of inflammation they manifest at the ultrastructural level in aerohematic barrier cells. The period of granuloma formation is characterized by cell hypertrophy, particularly of type II alveolocytes, intensive macrophagal reaction, and increased functional activity of more and more cells. Ultrastructural and functional differences in epithelioid and multinuclear giant cells are revealed. A progressive course of tuberculosis involves decompensation of the compensatory and repair processes. Contrary to this, during chemotherapy of tuberculosis repair reactions predominate, resolution of inflammatory changes is in progress, cell populations in the granuloma change, and it is separated from the adjacent pulmonary tissue.

Enhanced production of TGF-beta by blood monocytes from patients with active tuberculosis and presence of TGF-beta in tuberculous granulomatous lung lesions.

The expression of TGF-beta, a molecule that affects both immune responsiveness and wound healing, was examined in blood monocytes and granulomatous lesions from patients with active pulmonary tuberculosis. The spontaneous release of TGF-beta was higher in culture supernatants of monocytes from patients as compared with those of healthy subjects by an ELISA (p < 0.0005). TGF-beta activity was also confirmed in a bioassay in supernatants from patients. Next, freshly isolated monocytes from patients with tuberculosis and matched subjects were examined for TGF-beta activity. Cytosmears of monocytes were stained with an Ab against TGF-beta 1 (anti-LC) or isotype-specific Ab by using an alkaline-phosphatase anti-alkaline phosphatase method. In contrast to monocytes from healthy individuals, 60 to 70% of monocytes from patients demonstrated cytoplasmic staining for TGF-beta (n = 3). Upon hypotonic lysis, monocytes from patients with tuberculosis contained immunoreactive TGF-beta (n = 3). By Northern blot analysis, monocytes from three of seven patients with tuberculosis had increased expression of TGF-beta mRNA as compared with concurrently examined monocytes from healthy subjects. Within the granulomas of lung sections from two patients with untreated tuberculosis, TGF-beta immunoreactivity was identified in the Langhan's giant cells mainly and to a lesser extent the epithelioid cells using anti-LC Ab and the peroxidase-anti-peroxidase technique. Thus, both blood monocytes and lung granuloma macrophages from patients with active tuberculosis express TGF-beta. Excess activity of this cytokine in blood monocytes may underlie the depressed T cell responses of patients with tuberculosis. Moreover, within the infected tissues excess TGF-beta activity may interfere with anti-mycobacterial mechanisms and effective granuloma formation.

The study of histamine H1- and H2-receptors in human lung cancer.

Data on human lung histamine H1- and H2-receptors in cancer and chronic inflammatory processes are reported. It has been found that the number of histamine H1-receptors significantly increases both in cancer and chronic pneumonia and does not practically change in tuberculosis lung parenchyma. The binding parameters of histamine H2-receptors both in cancer and inflammatory processes were similar to those obtained for the normal tissue. The important role of parenchymal histamine H1-receptors in the neuromodulation of airways in human lung adenocarcinoma is discussed.

[Changes in the granuloma in the experimental treatment of tuberculosis with a prolonged form of isoniazid]. / Izmeneniia granulemy pri lechenii tuberkuleza prolongirovannoi formoi izoniazida v éksperimente.

The main difficulties encountered in the treatment of the persistent forms of a chronic tuberculous process are associated with a difficult access to mycobacteria persisting in the vacuolar apparatus of macrophages that form granulomas as well as with a high level of toxic affections of the parenchymatous liver cells. The complex long-acting isoniazid preparation on the dextran basis and the scheme of its application prevented, to a considerable degree, the above complications under the experimental conditions and hence can be considered as promising for clinical use.

[Muscarinic acetylcholine receptors in peripheral human lung tissue: binding characteristics of 3H-quinuclidinyl-benzilate in malignancies]. / Muskarinovye atsetilkholinovye retseptory v perifericheskoi tkani legkikh cheloveka: kharkteristika sviazyvaniia 3H-kvinuklidinil-benzilata pri zlokachestvennykh novoobrazovaniiakh.

Binding of muscarinic receptors antagonist 3H-quinuclidinyl benzylate (3H-QNB) was studied in human peripheral lung tissue, obtained after resection of pulmonary carcinoma and partial resection of tuberculoma from surrounding normal tissues (control group). Radioligand binding assays showed that binding of 3H-QNB in lung parenchyma membranes of both normal, healthy persons and patients with cancer was saturable with high rate of affinity. Saturation occurred at 1 nm concentration of the ligand. The Scatchard plot analysis indicated that the high affinity binding site exhibited the following parameters: Kd = 0.33 nm, Bmax = 164.8 fmol/mg in the group of controls: and Kd = 0.63 nm, Bmax = 401.5 fmol/mg in the group of patients with cancer. Maximal amount of 3H-QNB binding sites was distinctly increased in malignization of peripheral lung tissue. The results obtained suggest that increase in concentration of 3H-QNB binding sites may be among the specific patterns of malignant growth in lung peripheral tissue.

[The DNA content in precancer, peripheral cancer and carcinoids of the lung (histospectrophotometric research)]. / Soderzhanie DNK pri predrake, perifericheskom rake i kartsinoidakh legkogo (gistospektrofotometricheskoe issledovanie).

Histospectrophotometric study of DNA content shows that, at the lung periphery, oval structures with atypia, adenomatosis with atypia, dysplasia and basal cell hyperplasia with bronchial epithelium atypia should be regarded as precancerous lesions. Carcinoma in situ and bronchoalveolar carcinoma are distinguished from the foci of epithelial dysplasia by an increase of DNA content and appearance of hetero- and aneuploidy. Aneuploidy is observed in 97% peripheral lung carcinoma and is combined with a heterogeneity of cells by their DNA content. Poorly differentiated squamous cell carcinoma, adenocarcinoma and large cell carcinoma are characterized by a marked heterogeneity which correlates with a degree of tumour morphological atypia. Bronchoalveolar carcinoma is a heterogeneous group of tumours with a different histogenesis and pronounced anaplasia which is expressed in a various degree of polyaneuploidy and heteroploidy. Small cell lung carcinoma, a special group of tumours, which, as distinct from other carcinoma types, is characterized by a low, sometimes diploid DNA content which does not correlate with its malignancy.