EXTRACTION OF FERRUGINOUS BODIES FROM LUNG TISSUE OBTAINED AT SURGERY and AUTOPSY

Ferruginous bodies were extracted quantitatively from 508 patients; operated 242 with lung cancer and 94 with non-lung cancer, and autopsy 51 from hepatomas, 42 from stomach cancers, and 79 from non-cancer diseases. The bodies were divided morphologically into 4 types; 2 types may be from asbestos and the other 2 types from carbon and silicate which were very rarely found. The incidence of the bodies was the highest in hepatoma 96.8, followed by lung cancer 90.8%, non-lung cancer 80%, stomach cancer 76.2%, and non-cancer 74.2% in the years 1975–1981. Distribution of count of asbestos bodies was characteristic in patients with lung cancer, i.e. the cases who had asbestos bodies above 100/5 g of wet lung were found in 36 of 242 patients with lung cancer (14.8%) who were all smokers and 15 of 266 patients with the other diseases (5.6%) with significant difference between the two groups. Moreover, out of 14 patients who had the bodies above 300/5 g of wet lung, 9 were patients with lung cancer and also smoked heavily, and remaining 5 patients with diseases other than lung cancer consisted of 2 heavy smokers, a moderate and a mild smoker, and a non-smoker. These evidences may suggest the existence of some relation between occurrence of lung cancer and low degree of asbestos exposure with addition of smoking.     

Frequency of "blue bodies" in pulmonary cytology specimens

To determine the frequency of occurrence of "blue bodies" (BBs), 2,010 pulmonary cytology specimens (1,403 sputum and 607 bronchial specimens) from 1985 were reviewed. The smears were examined microscopically by transmitted and polarized light. BBs were extracellular structures, occurring most commonly in clusters but sometimes conglomerated. With Papanicolaou stain, they had a characteristic birefringent laminated brown core and a nonbirefringent blue rim. Chemical microanalysis and energy-dispersive x-ray analysis revealed that their chemical composition was calcium carbonate. A total of 233 specimens contained BBs, with a frequency of 10.5% in bronchial specimens and 12% in sputum. Only 8.6% of the BBs had co-existent pulmonary malignancy. We concluded that BBs were common structures in pulmonary cytology and were not associated with pulmonary malignancy or pulmonary fibrosis in our series. They must be distinguished from contaminants, staining artifacts, and parasitic ova.     

The role of persisting infections in the pathogenesis of pulmonary emphysema. Electron microscopy reveals a probable bacterial colonization of the alveolar space and the bronchioles

Lung volume reduction surgery (LVRS) yields resection specimens from patients with advanced pulmonary emphysema. Regarding the development of lung function parameters, recent results obtained by light microscopy revealed an unfavorable prognosis in patients with remarkable inflammation, particularly in the bronchioli. Tissue from ten patients (alpha1-antitrypsin level in the normal range) was furthermore investigated by electron microscopy. Scanning electron microscopy shows 0.4-0.6 micron spherical bodies variably densely arranged in the whole alveolar space and in the bronchioles of all patients. These bodies are mostly seen on the microvilli of type II pneumocytes. An immunological reaction with activation of macrophages and granulocytes occurs simultaneously. Macrophages show cytoplasmic extensions to the spherical bodies, which exhibit a cellular membrane but no cellular wall. This favors the diagnosis of bacterial colonization of the alveolar space and the bronchioles by mycoplasmas or L-forms of other bacteria. As patients undergoing lung volume reduction surgery are under optimal medical treatment and without any infection clinically, these findings appear to be relevant for the pathogenesis and/or progression of pulmonary emphysema.     

Endothelial cell morphology in focal areas of in vivo Evans blue uptake in the young pig aorta. I. Quantitative light microscopic findings

The morphology of endothelial cells from the pig aorta was examined using silver nitrate-stained Häutchen preparations. Areas of enhanced uptake of the protein-binding azo dye Evans Blue (blue areas) and areas of no dye uptake (white areas) were compared. From representative light photomicrographs the appearance of silver granules, stigmata, stomata and gaps in the silver-stained cell boundaries was examined. The numbers of these structures per cell were counted in both blue and white areas, and cell length and polarity in the two areas were also quantitated.Silver-stained cell boundaries in blue areas appeared consistently thicker and more irregular than in white areas, and cells from the latter were often longer and more tapered. Quantitatively, a significantly greater number of gaps or breaks in the boundary lines was found in blue areas. Over 85% of cells from both areas were aligned within 30° of the long axis of the vessel, and stigmata and stomata occurred with the same frequency in both areas. These and other findings are discussed in terms of the known permeability differences between areas of Evans Blue uptake and areas of no dye uptake.     

Morphological findings at the carotid bodies of humans suffering from different types of systemic hypertension or severe lung diseases

Post-mortem studies of the carotid bodies of 62 humans were carried out using light microscopic and morphometric methods. According to the clinical and autopsic data the subjects were divided into 5 groups: normotensives, essential hypertensives, renal hypertensives, chronically hypoxic persons with severe lung diseases, and people who suffered from both lung diseases and essential hypertension. Carotid body volume showed age-dependence in the normotensive group; the biggest glomera carotici were found at an age of 40-60 years, whereas younger and older people exhibited smaller carotid bodies. In the group with the essential hypertensives only old patients exhibited enlarged carotid bodies. In younger essential hypertensives but also in the renal hypertensives an increase of carotid body size was not demonstrable. The people with severe lung diseases regularly had greater carotid bodies when compared with age-matched normotensive subjects. In addition, chronically hypoxic patients had a proliferation of type II cells, perhaps with involvement of Schwann cells and fibrocytes. This increases of elongated cells was only seldom observed in the other groups. The results are discussed with respect to the alterations known so far of arterial chemoreceptor function and reflex effects in systemic arterial hypertension.     

Asbestos fibers and pleural plaques in a general autopsy population.

It has been claimed that symmetric lower zone pleural or diaphargmatic plaques are markers of asbestos exposure both in asbestos workers and the general population. In this study, total pulmonary asbestos burden was analyzed for 29 patients selected because pleural plaques were found at autopsy, and the results were compared with values obtained for 25 patients who had no occupational asbestos exposure. The average number of asbestos bodies in the plaque groups was 1732/g wet lung, and in the control group, 42/g wet lung. Uncoated asbestos fibers were extracted from lung and counted, measured, and identified by morphologic examination, electron diffraction, and energy-dispersive x-ray spectroscopy. The total number of fibers/per gram wet lung in the plaque group (114 x 10(3)) was similar to that in the control group (99 x 10(3), as was the number of chrysotile fibers (51 x 10(3) versus 29 x 10(3)). However, the plaque patients had a marked increase in the number of the commercially used high aspect ratio amphiboles, amosite and crocidolite (50 x 10(3) versus 1 x 10(3). A retrospective history of fairly certain asbestos exposure was obtained for 16 of the plaque patients, and such a history correlated strongly with increased numbers of commercial amphiboles in lung. It is concluded that 1) in this general autopsy population, two subgroups of patients are present. About one half of the patients appear to have developed pleural plaques as a result of asbestos exposure, while the etiology of the plaques in the other half is unclear; 2) the presence of pleural plaques correlates with a modest (50-fold) increase in numbers of long high-aspect ratio commercial amphiboles in lung tissue but does not correlate with numbers of chrysotile fibers, noncommercial amphiboles, or the total number of asbestos fibers; 3) asbestos-induced lesions are related to a complex set of mineralogic parameters and not to mere numbers of fibers in lung.     

Amiodarone lung: pathologic findings in clinically toxic patients

Lung biopsy and autopsy specimens of 12 patients with amiodarone pulmonary toxicity were studied to better characterize the pathology of amiodarone lung. For comparison, the autopsy specimens of five patients taking amiodarone without pulmonary side effects also were examined. Interstitial pneumonia was the most common manifestation of amiodarone lung and was characterized by interstitial inflammation, fibrosis, and hyperplasia of type II pneumocytes. Hyaline membranes were present in two cases. Foamy alveolar macrophages were present in all but one patient, and in four associated organizing pneumonia was present. Foamy alveolar macrophages also were present in three of five clinically nontoxic patients. Electron microscopy demonstrated membrane-bound lamellar inclusions in all of the three cases of amiodarone lung examined. Inclusions also were present in two of five patients who died of other causes. The authors conclude that amiodarone lung is primarily an interstitial pneumonia. Foamy alveolar macrophages and cytoplasmic lamellar inclusions are characteristic, but neither is specific, and their presence alone does not distinguish toxic from nontoxic patients.     

Cytomegalovirus studies of autopsy tissue. II. Incidence of inclusion bodies and related pathologic data.

Cytomegaloviruses (CMV) were recovered from lung tissue in 34 (6.8%) of 502 unselected autopsy cases. Inclusion bodies were detected in the lung in nine of these cases (26%) and in organs other than the lung in three others. Overall, the incidence of inclusion bodies in this series of 502 cases was 2.4%. Our data strongly indicate that virus isolation is more sensitive than histopathologic study in establishing the presence of CMV infection. However, CMV was not recovered from one kidney and one liver in which inclusion bodies were present, although the virus was isolated from lung. Four of five cases of renal allograft rejection were positive for both CMV and inclusion bodies. The incidence of CMV recovery and inclusion body detection in leukemia and lymphoma cases was more than twice that in cases with other diseases. CMV inclusion bodies with or without associated inflammation were found, in descending order of frequency, in the lung, kidney, liver, pancreas, adrenal gland, esophagus, prostate, testes, thyroid gland, parathyroid gland, stomach, small intestine, large intestine, and heart.     

Pathological calcification in juvenile dermatomyositis (JDM): microCT and synchrotron x-ray diffraction reveal hydroxyapatite with varied microstructures

The objective of this study was to begin to relate the microstructure of calcinosis samples to clinical and laboratory characteristics of the juvenile dermatomyositis (JDM) patients. Laboratory x-ray microCT (micro-Computed Tomography) noninvasively mapped microstructure for the first time in JDM calcifications. Synchrotron x-ray diffraction (transmission geometry) identified the mineral phase and crystallite size in the deposits. Samples were obtained from four children who had active JDM longer than 80 months and who were typed for TNFalpha-308 allele polymorphisms. Uniform mineral (giving the appearance of an extruded solid) was observed in one patient, and irregular blocks of differing sizes filled the samples from two other patients. The sample from the fourth patient appeared to combine features of the other two types. These spatial distributions of mineral were quite different from those in a bone reference sample. The only mineral observed in the JDM samples was hydroxyapatite (HAP), and the diffraction peaks of the JDM samples were slightly narrower than those of a trabecular bone reference sample. Diffraction peak widths of the JDM specimens revealed crystallite sizes (approximately 220-240 A) that are comparable to values reported in the literature for bone. Three children were positive for TNFalpha-308 GA polymorphism. The data suggest several possible origins for blocky vs. uniform structure of the JDM calcifications, including differences in duration of untreated inflammation, in TNFalpha-308 polymorphism, and in mechanical constraint at the calcification site. Information from additional samples is required to determine the relative role of each of these factors. Taken together, non-invasive microCT and x-ray diffraction characterization on the same samples offer an informative window into the dystrophic mineralization process in JDM.     

Pseudomesotheliomatous adenocarcinoma: a reappraisal.

Adenocarcinomas of or in lung that clinically and pathologically mimic diffuse pleural mesotheliomas are rare. We reviewed selected clinical and pathologic features of 15 autopsy/surgical cases previously reported in the medical literature and of 15 additional cases from the files of the Armed Forces Institute of Pathology (AFIP). Ninety percent of the patients were men. The median age was 61 years. Sixty-three percent of the patients smoked, 17% of them had possible or definite occupational exposure to asbestos, and one patient had microscopically proven asbestosis. Most patients had chest pain, shortness of breath, or cough, and had unilateral pleural effusion in the chest x-ray. At thoracotomy or at autopsy, numerous nodules, plaques, or a continuous rind of tumor was present over the pleural surface. Microscopically, the tumors showed simplified glands, nests, cords, papillary, tubulopapillary or biphasic patterns of growth. The neoplasms contained mucin that stained with diastase-predigested periodic acid-Schiff (PAS), mucicarmine, and alcian blue (with or without hyaluronidase predigestion). All patients died with/of tumor, with a mean survival of 4.7 months for those reported in the medical literature and of 7 months for those in the AFIP files. These adenocarcinomas therefore mimic pleural mesothelioma not only in their clinical and gross and microscopic appearance, but also in their prognosis.     

Significance of crystalline inclusions in lung granulomas

We describe six examples of nonnecrotizing lung granulomatosis in which there were numerous polarizable crystalline inclusions. The crystals were easily visible in routine H&E-stained slides and were so prominent that the question of a pneumoconiosis or other exogenous source was raised. There was no clinical history to suggest an inhalational source, however, and no patient used intravenous drugs. In one case, an atypical mycobacterial infection was proven to be etiologic, while sarcoidosis was documented in three. A review of 63 additional consecutive lung biopsies and 24 extrapulmonary biopsies showing nonnecrotizing granulomatous inflammation demonstrated crystals in almost two-thirds of cases. X-ray spectroscopy and histochemistry demonstrated that the crystals contained mainly calcium oxalate and calcium carbonate and thus represented products of cellular metabolism. These findings emphasize that crystalline inclusions are common in lung granulomas of varying etiology. They may be numerous, and their presence does not necessarily indicate a pneumoconiosis or other exogenous source.     

Photoconversion of fluorescent retrograde tracers.

Photoconversion of fluorescent dyes, retrogradely transported through axons to their parent cell bodies, into a stable diaminobenzidine (DAB) reaction product was tested in the nigrostriatal and thalamocortical systems of rats. Satisfactory results were obtained with Propidium Iodide (PI), Fluoro-Gold (FG), Fast Blue (FB), Diamidino Yellow (DY), and rhodamine-labeled latex microspheres (RLM); some photoconversion was also observed in Evans Blue (EB)-labeled neurons. The red fluorescent tracers PI, EB and RLM were photoconverted under the excitation wavelength appropriate for eliciting their fluorescent emission. With the yellow or blue fluorescent tracers FG, FB, and DY satisfactory results could instead be obtained using an excitation wavelength which did not elicit visible fluorescent emission. This finding indicates that the latter is not a critical factor for obtaining photoconversion. The features and subcellular localization of photoconverted DAB were different from those of the fluorescent labeling: photoconversion resulted in the appearance of brown granules of DAB reaction products in the cytoplasm, independently from the occurrence of fluorescent labeling in the neuronal cytoplasm or nucleus. Photoconversion may enable new applications of fluorescent retrograde tracing and, in particular, its electron microscopic visualization.     

Immunohistochemical localization of serotonin- and substance P-containing fibers around respiratory muscle motoneurons in the nucleus ambiguus of the cat

Retrograde tracing with a fluorescent dye (Fast Blue) combined with immunohistochemistry was used to determine if the putative neurotransmitters, serotonin and substance P, are present around posterior cricoarytenoid muscle motoneurons. Fast Blue was injected into the posterior cricoarytenoid muscle of the larynx. Following a 14-21 day survival time to allow for transport of the dye, the animals were perfusion fixed and the brainstem was removed for analysis under the fluorescence microscope. Retrogradely labeled cell bodies containing Fast Blue were found within the nucleus ambiguus from 0.5 to 3.0 mm rostral to obex. These motoneurons ranged in size from 23 to 38 micron. The same tissue sections containing labeled posterior cricoarytenoid muscle motoneurons were then used to determine the distribution of serotonin and substance P around these motoneurons using the indirect immunofluorescence technique. A dense network of serotonin-containing immunoreactive fibers was found around posterior cricoarytenoid muscle motoneurons. The fibers contained varicosities which were in close proximity, actually appearing to surround these motoneurons. Substance P immunoreactive fibers and varicosities were also found around posterior cricoarytenoid muscle motoneurons. The density and pattern of distribution of the substance P immunoreactivity was similar to that of the serotonin immunoreactivity. These results suggest that these putative neurotransmitters may be involved in influencing the activity of posterior cricoarytenoid muscle motoneurons. Serotonin and substance P are also present around other respiratory motoneurons such as phrenic motoneurons. Therefore, these two neurotransmitters may have a more general role in influencing respiratory motor outflow.     

Bronchiolitis interstitial pneumonitis: a pathologic study of 31 lung biopsies with features intermediate between bronchiolitis obliterans organizing pneumonia and usual interstitial pneumonitis, with clinical correlation

Bronchiolitis combined with interstitial pneumonitis generally has been equated with bronchiolitis obliterans organizing pneumonia (BOOP). We describe our experience with lung biopsies that had both bronchiolar and interstitial diseases. We studied 31 patients who had respiratory difficulty leading to open lung biopsy, which showed a combination of both prominent bronchiolitis and prominent interstitial pneumonitis. We compared these cases clinically and pathologically with 6 other pulmonary diseases, namely, bronchiolitis obliterans, BOOP, nonspecific interstitial pneumonitis, usual interstitial pneumonitis, airway-centered interstitial fibrosis, and idiopathic bronchiolocentric interstitial pneumonia, and with 10 cases of cystic fibrosis, an unrelated disease with both bronchiolar and interstitial pathology. The commonality of our cases was a combination of bronchiolitis and interstitial inflammation and fibrosis but little or no intra-alveolar organizing pneumonia. Bronchiolitis obliterans with organizing pneumonia involved less area than the interstitial pneumonitis in each case. All 19 patients for whom we had follow-up received corticosteroids for their pulmonary diseases. Seven patients had improvement in symptoms and pulmonary function test results and radiographic findings, 5 patients experienced subjective improvement with unchanged results of pulmonary function tests or chest x-ray, 1 patient's condition was unchanged, 6 patients' disease worsened, and 4 of these 6 died. The natural history of these cases, which we have designated bronchiolitis interstitial pneumonitis, seems more sanguine than usual interstitial pneumonitis and worse than BOOP at least in the short term. On the one hand, response to corticosteroids was not as frequent as generally accepted for BOOP. On the other hand, disease did not progress in most patients on corticosteroids.     

The histological diagnosis of clinically documented cases of cryptogenic organizing pneumonia: diagnostic features in transbronchial biopsies

Eleven cases of clinically diagnosed cryptogenic organizing pneumonia were examined in order to establish the histological features found at transbronchial biopsy and to correlate this with open lung biopsy which followed in six cases. The essential pathological feature was the presence of buds of granulation tissue (Masson bodies) indicating organization of a persistent exudate by fibroblasts and capillaries within alveoli, with preservation of the alveolar architecture. In addition, both acute and chronic inflammatory cells were present in the interstitium. These features, combined with the clinical history, were sufficient for a diagnosis in seven of the 11 cases on transbronchial biopsies. Four biopsies lacked these features. Patients proceeded to open lung biopsy in addition to the two with histological features of cryptogenic organizing pneumonia on transbronchial biopsy, but where the clinician wanted to eliminate other pathology because of rapid clinical deterioration. Five of the six cases coming to open lung biopsy confirmed cryptogenic organizing pneumonia with Masson bodies within alveoli, but changes were focal in three with very few Masson bodies in one. One case which had the features on transbronchial biopsy lacked them in the open lung biopsy. Transbronchial biopsy, therefore, can yield diagnostic material in the majority of patients with cryptogenic organizing pneumonia while open lung biopsy, which is considered the gold standard for interstitial lung disease, may yield negative results because of sampling error and the rapid evolution and changing pattern of the disease.     

Pathological Calcification in Juvenile Dermatomyositis (JDM): MicroCT and Synchrotron X-Ray Diffraction Reveal Hydroxyapatite with Varied Microstructures

The objective of this study was to begin to relate the microstructure of calcinosis samples to clinical and laboratory characteristics of the juvenile dermatomyositis (JDM) patients. Laboratory x-ray microCT (micro-Computed Tomography) noninvasively mapped microstructure for the first time in JDM calcifications. Synchrotron x-ray diffraction (transmission geometry) identified the mineral phase and crystallite size in the deposits. Samples were obtained from four children who had active JDM longer than 80 months and who were typed for TNFα -308 allele polymorphisms. Uniform mineral (giving the appearance of an extruded solid) was observed in one patient, and irregular blocks of differing sizes filled the samples from two other patients. The sample from the fourth patient appeared to combine features of the other two types. These spatial distributions of mineral were quite different from those in a bone reference sample. The only mineral observed in the JDM samples was hydroxyapatite (HAP), and the diffraction peaks of the JDM samples were slightly narrower than those of a trabecular bone reference sample. Diffraction peak widths of the JDM specimens revealed crystallite sizes (～ 220–240 Åa) that are comparable to values reported in the literature for bone. Three children were positive for TNFα -308 GA polymorphism. The data suggest several possible origins for blocky vs. uniform structure of the JDM calcifications, including differences in duration of untreated inflammation, in TNFα -308 polymorphism, and in mechanical constraint at the calcification site. Information from additional samples is required to determine the relative role of each of these factors. Taken together, non-invasive microCT and x-ray diffraction characterization on the same samples offer an informative window into the dystrophic mineralization process in JDM.     

Absence of tubular myelin in lungs of infants dying with hyaline membrane disease.

Immaturity of the pulmonary surface active material synthesizing system with deficiency of surface active material in the premature lung is an accepted cause of hyaline membrane disease. Lamellar bodies, the intracellular form of surface active material, are produced and secreted from Type II pneumocytes and converted to tubular myelin in the alveolar lumen. Tubular myelin, in turn, gives rise to the surface monolayer, which has the greatest surface active property. Thus, lung sections were studied by light and electron microscopy from 35 infants who died of histologically confirmed hyaline membrane disease and 19 infants who died of other causes. Tubular myelin was not identified ultrastructurally in lungs of infants who died of hyaline membrane disease, despite the presence of abundant lamellar bodies. In contrast, 16 of 19 infants dying of other causes had easily identifiable tubular myelin in addition to lamellar bodies. The absence of tubular myelin in the hyaline membrane disease patients suggests an abnormality in the conversion of lamellar bodies to tubular myelin. The authors speculate that this abnormal lamellar body turnover may be important in the pathogenesis of hyaline membrane disease.     

Spinal projections of hypothalamic histidine decarboxylaseimmunoreactive neurones

The existence of a histidine decarboxylase (HDC)-immunoreactive diencephalo-spinal pathway in the rat was demonstrated using an antiserum raised against HDC from fetal rat liver. HDC-immunoreactive nerve cell bodies were numerous in the ventral and lateral caudal hypothalamus. More caudally, in the mesencephalon, no cell bodies were observed but fairly many, transversely cut nerve fibres, were found in association with the fasiculus longitudinalis medialis bilaterally. At the most caudal medullary level, these longitudinally passing fibres became displaced ventrally to a position just laterally to the pyramidal decussation. In the spinal cord the fibres were more dispersed and rather sparse in most areas. The existence of a diencephalo-spinal HDC-immunoreactive pathway was verified by analyzing material from rats which had received injections of the retrograde fluorescent tracer True Blue into the cervical spinal cord. True Blue fluorescence and HDC immunofluorescence were found to coexist in a subpopulation of the HDC-immunoreactive neurones in the hypothalamus.     

Infection-induced airway fibrosis in two rat strains with differential susceptibility.

Chronic infections play a significant role in the morbidity and mortality of patients with chronic airflow limitation. By stimulating airway inflammation, persistent infection has the potential to cause airway fibrosis. However, in patient this condition is most typically found in lungs damaged by other factors, such as smoking, abnormal secretions, or barotrauma. We report the characterization of Mycoplasma pulmonis infection-induced lung fibrosis in two immunocompetent rat strains with no preexisting lung disease. The fibrosis was predominantly in the airways, as demonstrated by the findings for infected animals of increased airway inflammation, airway fibrosis, and airway wall thickness, which correlated with the collagen content of the lungs. Also, the physiological alterations were the opposite of those found in interstitial fibrosis, with a positive correlation between lung compliance and collagen content. The airway fibrosis was noted earlier and to a greater extent in Lewis rats than in Fisher rats, and this result apparently was related to regulation of the inflammatory response. Airway wall thickness, airway inflammation, and airway fibrosis are commonly reported in tissue specimens from patients with chronic airway diseases and have been shown to correlate with airflow limitation in patients with chronic obstructive pulmonary disease. Thus, this model may be useful in furthering our understanding of the role of chronic infection and airway inflammation in airflow obstruction.