Epidemiology of sarcoidosis in Japan.

The present study was designed to identify recent clinical phenotypes using the National Epidemiological Survey and to compare findings with those of previous surveys in Japan. Pathologically confirmed sarcoidosis cases newly diagnosed in 2004 were eligible for the present study. Disease parameters were recorded and compared. A total of 1,027 patients were enrolled from a cluster encompassing 79.4% of the entire Japanese population. The study participants consisted of 364 males and 663 females, providing an average incidence rate of 1.01 per 100,000 inhabitants (0.73 for males and 1.28 for females). The age-specific incidence rate displayed a biphasic pattern in the whole patient population and in the females. The male incidence rates peaked in the 20-34-yr-old group. A second peak for 50-60-yr-old females showed a higher incidence than the first younger peak. Patients with abnormalities in eyes, skin and cardiac laboratory findings accounted for 54.8, 35.4 and 23.0% of cases, respectively. The female/male incidence ratio was increased, and the frequency of eye and skin involvement and cardiac abnormality was higher than in previous surveys conducted in Japan. In conclusion, the data obtained in the present study differ from those of other countries and showed changes in sarcoidosis clinical phenotypes compared with previous studies in Japan.

Phosphane Sulfide/Octacarbonyldicobalt-Catalyzed Pauson - Khand Reaction Under an Atmospheric Pressure of Carbon Monoxide.

Convenient access to cyclopentenones is provided by catalytic intra- and intermolecular Pauson - Khand reactions in the presence of octacarbonyldicobalt and tributylphosphane sulfide (see scheme). In contrast to other reactions of this type, they proceed under mild conditions (70 degrees C, 1 atm CO), and commercially available [Co(2)(CO)(8)] can be used without further purification.

Markers of cell proliferation and expression of melanosomal antigen in lymphangioleiomyomatosis.

Pulmonary lymphangioleiomyomatosis (LAM), a disease of young women, is characterized by proliferation of immature-appearing smooth-muscle cells (LAM cells) in the lungs and abdomen. LAM cells react with monoclonal antibody HMB45, which recognizes a 100-kD glycoprotein (gp100) originally found in human melanoma cells. We investigated the expression and the subcellular localization of gp100 in lung tissue from patients with LAM and in human melanoma cell lines (Malme-3M, A2058, and CHL-1), and the relationship between this expression and cellular proliferation. Binding sites for HMB45 antibody in melanoma and LAM cells were located in cytoplasmic granules resembling immature melanosomes. LAM cells reactive for proliferating-cell nuclear antigen (PCNA), a marker of cellular proliferation, were spindle-shaped, in contrast to the large, epithelioid cells reacting with HMB45 antibody. In accord with this finding, we observed an inverse relationship between the immunostaining for HMB45 antibody and PCNA in LAM and melanoma cells. Thus, LAM and melanoma cells are heterogeneous with respect to their stages of proliferation and their expression of melanoma antigens. PCNA-positive cells, which are more likely to be negative for reactivity with HMB45 antibody, may be more relevant to the progression of LAM than are HMB45-positive cells, which are the hallmark of LAM.

Selective expression of RT6 superfamily in human bronchial epithelial cells.

RT6 proteins are glycosylphosphatidylinositol (GPI)-linked alloantigens that are localized to cytotoxic T lymphocytes and that have nicotinamide adenine dinucleotide glycohydrolase and adenosine diphosphate (ADP)-ribosyltransferase activities. In view of the importance of GPI-linked surface proteins in mediating interactions of cells with their milieu, and the varied functions of airway cells in inflammation, we undertook the present study to determine whether human homologues of the RT6 superfamily of ADP-ribosyltransferases (ART) are expressed in pulmonary epithelial cells. We hypothesized that these surface proteins or related family members may be present in cells that interact with inflammatory cells, and that they may thereby be involved in intercellular signaling. Using in situ analysis and Northern blot analysis, we identified ART1 messenger RNA (mRNA) in airway epithelial cells. As expected for GPI-anchored proteins, the localization of ART1 at the apical surface of ciliated epithelial cells was demonstrated by staining with polyclonal anti-ART1 antibody, and was confirmed by loss of this immunoreactivity after treatment with phosphatidylinositol-specific phospholipase C (PI-PLC), which selectively cleaves GPI anchors and releases proteins from the plasma membrane. Using in situ hybridization with specific ART3 and ART4 oligonucleotides, we also identified two additional members of the RT6 superfamily in epithelial cells. In accord with these findings, we identified ART3 and ART4 mRNAs through reverse transcription- polymerase chain reaction of polyadenine-positive RNA from human trachea. Interestingly, these proteins appeared to be preferentially localized to the airway epithelium. The localized expression of these members of the RT6 superfamily in human pulmonary epithelial cells may reflect a role for them in cell-cell signaling during immune responses within the airway.

Comprehensive evaluation of 35 patients with lymphangioleiomyomatosis.

OBJECTIVES: To evaluate comprehensively the characteristics of lymphangioleiomyomatosis (LAM), with emphasis on the application of imaging and immunohistochemical methods. DESIGN: Prospective study. PATIENTS: Thirty-five female subjects with LAM. SETTING: Clinical Center, National Institutes of Health. INTERVENTIONS: BAL, pulmonary function test, ventilation/perfusion lung scans, CT of the chest and abdomen, ultrasonography of abdomen, and immunohistochemical study of lung biopsy specimens. RESULTS: Most patients had exertional dyspnea (83%) and pneumothorax (69%). BAL did not show diagnostic changes. The most common abnormalities on pulmonary function tests were decreased diffusing capacity of carbon monoxide (83%), hypoxemia (57%), and airway obstruction (51%). Bronchodilator response was found in 26% of patients. CT, which is almost pathognomonic, showed numerous thin-walled cysts throughout both lungs in all patients. Thirty-four patients (97%) had abnormal ventilation and/or perfusion lung scans. An unusual "speckling" pattern was observed on ventilation scans of 74% of patients. Common extrapulmonary features were retroperitoneal adenopathy (77%) and renal angiomyolipomas (60%). The percentage of abnormal smooth muscle cells (LAM cells), reactive with HMB45, varied from 17 to 67% in 10 lung biopsy specimens. CONCLUSIONS: Improved diagnostic methods have defined the abnormalities in patients with pulmonary LAM and increased the potential for early recognition and treatment of this disorder. Patients with LAM should be evaluated for bronchodilator responsiveness and may benefit from a trial of bronchodilators.

Immunohistochemical analysis of proteins of the Bcl-2 family in pulmonary lymphangioleiomyomatosis: association of Bcl-2 expression with hormone receptor status.

BACKGROUND: Pulmonary lymphangioleiomyomatosis (LAM), a disease of young women, is characterized by proliferation of abnormal smooth muscle cells (LAM cells), which often differ from normal pulmonary smooth muscle cells by frequently having estrogen and progesterone receptors. OBJECTIVE: To evaluate the relationships among several factors related to proliferation and apoptosis in LAM cells, we employed immunohistochemical methods for the localization of Bcl-2 and Mcl-1 (inhibitors of apoptosis), Bax (a promoter of apoptosis), c-Myc (an apoptosis-related oncoprotein), proliferating cell nuclear antigen (an indicator of mitotic activity), and nick end labeling (to identify apoptotic cells) in lung tissues of 9 patients with LAM. RESULTS: In all patients, most LAM cells reacted positively for Bax. The LAM cells were positive for both Bcl-2 and estrogen receptor in 5 patients, positive for only Bcl-2 in 1 patient, positive for only estrogen receptor in another patient, and negative for both in 2 patients. More than 50% of the Bcl-2-positive LAM cells were also positive for estrogen receptor. The reaction for c-Myc was positive in all patients. The immunoreactivity for Bcl-2 and Mcl-1, which inhibit apoptosis, was more intense in LAM cells than in normal vascular and bronchial smooth muscle cells. In 6 patients, more than 50% of the LAM cells were positive for proliferating cell nuclear antigen. Apoptosis was infrequent in LAM cells. CONCLUSIONS: Our results suggest that the expression of Bcl-2 in LAM cells may be related to hormonal regulation, and that by decreasing apoptosis, Bcl-2 and related proteins contribute to the imbalance between proliferation and death of LAM cells.

Clearance of mucus from endotracheal tubes during intratracheal pulmonary ventilation.

BACKGROUND: Intratracheal pulmonary ventilation (ITPV) is a form of tracheal gas insufflation in which all gas emerges in a cephalad direction from the tip of a reverse-thrust catheter positioned within an endotracheal tube. In vitro experiments have shown that this rapid gas flow, with 5 ml/h of normal saline added to the gas flow, continuously removes tracheal secretions from within the endotracheal tube. The authors evaluated its effectiveness to remove mucus in long-term studies in sheep. METHODS: Fourteen healthy sheep were tracheally intubated and ventilated for 3 days with ITPV or with volume-controlled ventilation. Measurements were made of the total amount of secretions within the endotracheal tubes (weight gain), the protein content within the endotracheal tubes, and the increase in resistance to constant air flow. The structure of the airways was examined grossly and histologically. Three additional sheep were ventilated for 24 h with ITPV, and Evans Blue dye was added to the saline to assess the distribution of the infused saline. RESULTS: There was significantly less mucus in endotracheal tubes of sheep ventilated with ITPV than with conventional ventilation, as shown by minimal weight gain (0.70 +/- 0.14 g vs. 2.44 +/- 0.81 g; P < 0.001), lower protein content (14.09 +/- 10.79 mg vs. 294.99 +/- 153.06 mg; P < 0.001), and lower resistance to constant air flow (6.15 +/- 0.54 cm H2O x 1(-1) x s(-1) vs. 15.34 +/- 5.28 cm H2O x 1(-1) x s(-1); P < 0.001). Results of gross and histological examinations of the tracheas of animals in both groups were similar, and the tracheas were well preserved. More than 95% of the instilled saline was recovered during ITPV. Only traces of Evans Blue dye were found near the tip of the endotracheal tubes. CONCLUSION: Intratracheal pulmonary ventilation makes it possible to keep the endotracheal tubes of sheep ventilated for 3 days free of mucus without suctioning.

Evolution of three patterns of intra-alveolar fibrosis produced by bleomycin in rats.

In pulmonary fibrosis, it is known that fibrotic changes develop in the intra-alveolar spaces and that intra-alveolar fibrosis can be classified into three patterns, namely intra-alveolar buds, mural incorporation and obliterative changes. In order to clarify the evolution of intra-alveolar fibrosis, immunohistochemical studies of extracellular matrix proteins and electron microscopic observations were made of the lungs of rats given a single intratracheal instillation of bleomycin. All three patterns of fibrosis developed in this model. Intra-alveolar buds changed into globular lesions with dense collagen deposition, the surface of which was covered by alveolar epithelium. Electron microscopy revealed that the buds often contained spiraling collagen fibrils and numerous microfibrils, but not mature elastic fibres, beneath the regenerating epithelial lining cells; the epithelial basement membranes were discontinuous. In contrast, mural incorporation and obliterative changes were associated with alveolar structural remodeling. Electron microscopically, these lesions had bundles of normal collagen fibrils, small elastic fibers, and continuous epithelial basement membranes. These results indicate that: (i) intra-alveolar buds, that become intra-alveolar collagen globules, with an unusual extracellular matrix, do not contribute to alveolar structural remodelling; and (ii) areas of mural incorporation and obliterative changes have the usual type of extracellular matrix and are essential for alveolar structural remodelling.

[Giant cell interstitial pneumonia in a metal grinder with an abnormally high level of serum CA19-9].

Interstitial pneumonia and recurrent pneumothorax developed in a 48-year-old man who had worked as a metal grinder. He died of respiratory failure despite having received antibiotics and steroids, and despite having undergone pleural sclerosis therapy. Giant cell interstitial pneumonia was diagnosed; innumerable bizarre giant cells engulfing black granules were found within the alveoli. The results of high-energy dispersion X-ray microanalysis indicated that the patient had hard metal pneumoconiosis associated with tungsten in the black granules. When he was admitted to the hospital, his serum CA19-9 and SLEX concentrations were abnormally high (2600 and 200 ng/ml, respectively). Immunohistochemical analysis of lung tissue was done with anti-CA19-9 and SLEX antibodies. CA19-9 staining revealed strong bronchialization and squamous metaplasia in contrast to type II hyperplasia. SLEX staining showed strong type II hyperplasia. Further investigations will be needed to determine the mechanism of elevated tumor-associated carbohydrate antigens in serum.

Serum neuron specific enolase level as a prognostic factor in non-small cell lung cancer.

In 93 patients with inoperable non-small cell lung carcinoma who underwent chemotherapy including cisplatin, the prognostic value of 9 factors were determined using Cox's proportional hazard model. Univariate analysis revealed that patients with a performance status of grade 2 (p < 0.01) or 3 (p < 0.05), those with stage IV disease (p < 0.05), those with a serum neuron specific enolase (NSE) level > 7.0 ng/ml (p < 0.001), and those with a low serum albumin level (p < 0.05) had a significantly worse prognosis. Multivariate analysis showed that a performance status of 2 or 3 and a high NSE serum level were associated with a significantly worse prognosis. More attention should be paid to the serum NSE level in patients with non-small cell lung carcinoma, because it not only reflects the tumor volume, but is also a prognostic factor which is dependent on individual tumor characteristics.

[Adhesion ultrastructures of mononuclear cells in experimentally-induced silicotic granuloma].

Experimental silicosis was induced by intratracheal infusions of 1 ml saline containing 50 mg standard silica (less than 5 microns diameter) in Sprague-Dawley rats. The lung tissues were observed histologically and ultrastructurally from half an hour up to 4 months. Macrophages, neutrophils, desquamated cells and their debris piled up around the alveolar ducts where the central cores of silicotic granuloma appeared. The granuloma became apparent by day 4 after the infusion and were covered by type II alveolar epithelial cells and bronchiolar cuboidal epithelial cells. Macrophages, fibroblasts and epithelial cells began to react to the antibody against proliferating cell nuclear antigen (PCNA) indicating self-replication on day 1. Macrophages in the granuloma made a close interdigitation with adjacent macrophages, and they gradually formed subplasmalemmal linear densities (SPLD) as paired forms between adjacent plasma membranes, and unpaired forms facing the interstitial matrix. SPLD were composed of linear densities with actin-like microfilaments along the leaflets of plasma membrane and were associated with extracellular dense bands which resembled a limited length of basement membrane. Interdigitation and SPLD structures were quite rare on day 1, but the number of macrophages with both structures increasingly appeared. The frequency of SPLD in macrophages also increased on a time course of granuloma maturation up to 4 months. Thus SPLD, which were originally found in the mononuclear phagocytes including macrophages, epithelioid cells and multi-nucleated giant cells, particularly in immune granuloma of man, also played a basic role in immobilizing macrophages in lesions of silica-induced granulomas.

[A case of pulmonary lymphangiomyomatosis induced by pregnancy].

A 35-year-old primigravida was admitted to the Department of obstetrics complaining of dyspnea and left back pain at 21 weeks' gestation. Chest roentgenogram revealed diffuse reticulonodular shadows predominantly in both lower lung fields and arterial hypoxemia was present. Pulmonary function tests showed restrictive impairment and decreased carbon monoxide diffuse capacity. From these results, interstitial pneumonia was suspected and she was first treated with prednisolone. However during her pregnancy, spontaneous pneumothorax occurred. Following spontaneous delivery of healthy infant at 37 weeks, left chylothorax occurred, and pleurodesis was performed with OK432. Thereafter the histological diagnosis of pulmonary lymphangiomyomatosis was made by transbronchial lung biopsy and treatment of prednisolone was stopped. She was treated with tamoxifen. In addition, progesterone-receptor was detected in the pulmonary tissue obtained at open lung biopsy. She was treated with cyclophosphamide in addition to tamoxifen. At present, shortness of breath has decreased slightly in comparison with one year previously, but no improvement has been seen in lung function tests or chest roentgenogram.