HRCT shows variations in appearance in disseminated tuberculosis in adults.

OBJECTIVE: To examine patterns of high resolution computed tomography (HRCT) of lungs of adults with disseminated tuberculosis (TB). DESIGN: Disseminated TB was defined as radiological involvement of all lung lobes. RESULTS: The case series illustrated wide variation in HRCT of disseminated TB. Patterns identified on HRCT included (1) miliary TB (haematogenous dissemination), (2) miliary TB with exudative reaction, (3) bronchogenic spread, (4) miliary TB mixed with bronchogenic spread, and (5) bronchogenic spread with multiple cavity formation. CONCLUSION: The HRCT patterns described allow classification of disseminated TB according to the mechanism of spread (haematogenous and/or bronchogenic) and the degree of local lung involvement (reaction or cavitation).

Mechanical agitation induces gene expression of NOR-1 and its closely related orphan nuclear receptors in leukemic cell lines.

NOR-1, NGFI-B and Nurr1 are closely related transcription factors that constitute a distinct subfamily within the nuclear receptor superfamily. Genes for these proteins are immediate-early genes, and are inducible in diverse cell types by various stimuli. In the present study, we investigated the effect of mechanical agitation on the gene expression of these transcription factors in cultured suspension cells by the quantitative reverse transcription-polymerase chain reaction. We found that mechanical agitation transiently induced NOR-1, NGFI-B and Nurr1 mRNAs in several leukemic cell lines in a dose-dependent manner. This induction was most pronounced in the HL-60 promyelocytic leukemia cell line, but also occurred to a lesser extent in other cell lines including KG-1, THP-1 and U937 cells. The induction was attenuated by serum or albumin, which are shear stress protectants for suspension culture cells. These reagents did not suppress forskolin-induced NOR-1 gene expression. These findings suggest the involvement of fluid shear stress in agitation-induced immediate-early gene expression. Since even moderate agitation could cause the induction, investigators should be cautious when evaluating the expression of immediate-early genes in some leukemic cell lines.

Expression of NOR-1 and its closely related members of the steroid/thyroid hormone receptor superfamily in human neuroblastoma cell lines.

Previously, we isolated a cDNA of neuron derived orphan receptor (NOR-1), a putative transcription factor with strong homologies to the orphan nuclear receptors NGFI-B and NURR1. In the present study, we examined the gene expression of NOR-1 as well as NGFI-B and NURR1 in human neuroblastoma cell lines by reverse transcription-polymerase chain reaction and nucleotide sequencing. Although the mRNAs of these orphan receptors were detected in all six neuroblastoma cell lines examined, basal expression levels of these genes varied among cell lines. Treatment with forskolin and 12-O-tetradecanoylphorbol-13-acetate rapidly increased the expression of all these genes in neuroblastoma NB-OK-1 cells. This induction did not require de novo protein synthesis, indicating that the NOR-1 gene as well as NGFI-B and NURR1 genes is an immediate-early gene. This is the first demonstration of NOR-1 gene expression in tumor cell lines.

Gene expression of NOR-1, a neuron-derived orphan receptor, is inducible in neuronal and other cell lineages in culture.

We recently isolated a cDNA of NOR-1, a novel member of the steroid/thyroid hormone receptor superfamily, from a rat neuronal cell cDNA library. NOR-1 has strong amino acid sequence homology to orphan receptors NGFI-B and Nurr1, and is expressed predominantly in the brain tissue in vivo. In the present study, we examined NOR-1 gene expression in various cultured cells with distinct properties of neuronal cells (PC12), glial cells (C6), endocrine cells (GH3, AtT-20 and Y-1), or fibroblasts (NIH3T3 and L), by reverse transcription-polymerase chain reaction and nucleotide sequencing. NOR-1 transcripts were induced with forskolin and 12-O-tetradecanoylphorbol-13-acetate in all cell lines examined. The nerve growth factor induced not only NGFI-B but also NOR-1 and Nurr1 transcripts in PC12 cells. These findings suggest that NOR-1 gene expression is induced by various stimuli in diverse cell types.

The NGFI-B subfamily of the nuclear receptor superfamily (review).

NGFI-B, Nurr1 and NOR-1 have structural features of ligand-activated transcriptional regulators and constitute the NGFI-B subfamily within the nuclear receptor superfamily. Since specific ligands for these molecules have not yet been identified, they are often called orphan nuclear receptors. Genes of the NGFI-B subfamily are classified as immediate-early genes that are induced rapidly but transiently by a variety of stimuli. Evidence accumulated over the past decade suggests this subfamily is involved in important physiological processes and cancer development. In this communication, we summarize and discuss their structural features, gene expression, physiological roles and oncological significance.

Secondary myeloid/natural killer cell precursor acute leukemia following essential thrombocythemia.

The de novo leukemic transformation of essential thrombocythemia is a rare event, and usually associated with previous treatments. We describe a patient who received treatments with nitrosourea for long-standing essential thrombocythemia and subsequently developed extramedullary tumors, tentatively diagnosed as lymphoblastic lymphoma. Combination chemotherapy was initially successful, but relapsed with marked bone marrow involvement. Surface marker analysis revealed that the tumor cells had CD5, CD7, CD33, CD34, and CD56 antigens but lacked other T-cell, and B-cell markers. Immunogenotypical studies revealed germline configurations for both T-cell receptors and immunoglobulin genes. These clinical and phenotypical features are consistent with a myeloid/natural killer cell precursor leukemia, a recently proposed distinct clinical entity. To our knowledge, this is the first report of secondary leukemia of myeloid/ natural killer cell precursor origin, and suggest that myeloid/natural killer cell precursor might be a potent target of therapy-related leukemia.

A case of leptomeningeal metastasis from lung adenocarcinoma diagnosed by reverse transcriptase-polymerase chain reaction for carcinoembryonic antigen.

A case of leptomeningeal metastasis from lung adenocarcinoma is reported. In this case, we evaluated the feasibility of reverse transcriptased polymerase chain reaction (RT-PCR) methods to detect cancer cells in cerebrospinal fluids (CSF). Messenger RNA of carcinoembryonic antigen (CEA) was clearly demonstrated in CSF by reverse RT-PCR methods. An immunohistochemical study also demonstrated that tumor cells were stained positive with anti-CEA antibody. This case suggests that RT-PCR for CEA was a sensitive and useful method to diagnose leptomeningeal metastasis from lung adenocarcinoma.

Expression of thyroid transcription factor-1 in 16 human lung cancer cell lines.

It has been suggested that thyroid transcription factor-1 (TTF-1) is frequently expressed in human lung cancer, especially in adenocarcinoma and small cell lung cancer, and the TTF-1 expression is closely related with the expression of surfactant protein. We hypothesized that TTF-1 is expressed in human lung cancer cell lines and its expression might be related to the expression of surfactant protein. To test this, expressions of TTF-1 and surfactant protein A (SP-A) were immunohistochemically evaluated in 16 human lung cancer cell lines. In addition, expressions of mRNAs for TTF-1 and SP-A were analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR) and sequencing. As a result, nuclear staining of TTF-1 was observed in two of six adenocarcinoma cell lines, none of seven small cell lung cancer cell lines, and none of three squamous lung cancer cell lines. Among the 16 cell lines, six cell lines (PC3, LC2/Ad, A549, RERF-LC-OK, HI1017, and PC9) expressed significant amounts of mRNA for TTF-1. In contrast, cytoplasmic staining of TTF-1 was observed in five of six adenocarcinoma cell lines, in six of seven small cell lung cancer cell lines, and in all three squamous cell lung cancer cell lines. One of the two adenocarcinoma cell lines those showed positive nuclear staining and cytoplasmic SP-A staining released a significant amount of SP-A in culture supernatant. Our present study demonstrates that the frequency of TTF-1 expression in the nucleus was very low in human lung cancer cell lines; however, their cytoplasmic positivities should be further investigated.

Cavitary lung cancer with an aspergilloma-like shadow.

A 66-year-old man who complained of cough and haemoptysis had a cavitary lesion with the meniscus sign in the right lower lung field on his chest X-ray and CT scan. He had smoked 40 cigarettes daily, for about 46 years. Initially, he was diagnosed with aspergilloma and given an antifungal agent. After 2 months, the cavitary lesion showed a slight irregularity of the inner border. The walls were irregularly thickened and were surrounded by infiltrative densities compared with the previous chest radiograph. Enlargement of right hilar and mediastinal lymph nodes was also observed. The fungus ball-like shadow was fixed on the anterior wall of the cavity and its position was not altered with the patient's movements. These radiographic findings led to suspicion that the lesion might be malignant. Transbronchial lung biopsy of the cavity wall and CT guided needle aspiration biopsy of the fungus ball-like lesion were performed. Microscopic examination revealed a squamous-cell carcinoma in both the cavity wall and the fungus ball-like lesion. There was no evidence of fungal elements. In conclusion, the meniscus sign is most often associated with benign diseases such as aspergilloma, however, one should remember that carcinoma may be a cause.

The point mutation in the promoter region and the single nucleotide polymorphism in exon 1 of the cytokeratin 19 gene in human lung cancer cell lines.

The CYFRA 21-1 assay which detects the cytokeratin 19 (CK19) fragment is widely used as a tumor marker for lung cancer. We previously suggested that the failure of PCR amplification of exon 1 is closely related to the inability of the expression of mRNA for CK19, and hypothesized that point mutations might exist within exon 1. In order to prove this, sequence analysis of the promoter region and exon 1 was performed in 14 human lung cancer cell lines. Among the 14 lung cancer cell lines evaluated, point mutations within the promoter region (at -99, G-->C) of the CK19 gene were demonstrated in two cell lines (Lu135 and HI1017). In addition, point mutations within exon 1 (at 90, T-->C, Ala-->Ala and at 179, G-->C, Gly-->Ala) were also demonstrated in three cell lines (LU135, HI1017, and LC2/AD). Point mutations within the promoter region of CK19 (at -99) and within exon 1 (at 179) were confirmed by analysis of digestion by specific restriction enzymes. Since the same point mutation within exon 1 (at 179) was observed in genomes of normal volunteers, this mutation was considered as a single nucleotide polymorphism. In contrast, there were no mutations within the promoter region of exon 1 in genomes of normal volunteers. After a computer search, it was demonstrated that several transcription factors bind to the sense primer sequence which was designed for amplification of exon 1. In addition, after point mutations within the promoter region occurred (at -99), new sequences appeared to which known transcription factors (AP2) bind. In conclusion, analysis of genomic DNA for CK19 suggested that expression of mRNA for CK19 was regulated by several transcription factors which bound to the specific sequence with the promoter region of the CK19 gene. It was also suggested that the mutation in the promoter region of the CK19 gene down-regulated the expression of mRNA for CK19.

Mechanisms of the release of CYFRA21-1 in human lung cancer cell lines.

The CYFRA 21-1 assay which detects cytokeratin 19 (CK19) fragment is widely used as a tumor marker for lung cancer. However, the reason why some lung cancer cell lines release CK19 fragment in culture supernatants and others do not, remains unclear. It was hypothesized that the release of CK19 fragment may be elucidated by the expression of mRNA for CK19. In order to prove this, the mRNA for CK19 was quantitatively evaluated by the competitive reverse transcriptase-polymerase chain reaction (competitive RT-PCR). The level of CYFRA 21-1 in the culture supernatant was measured by an immunoradiometric assay. CK19 protein synthesis was evaluated by a Western blotting and immunohistochemistry. Fourteen lung cancer cell lines were evaluated, and the amount of mRNA correlated well with the level of CYFRA 21-1 in culture supernatants. Analysis of genomic DNA for CK19 demonstrated that three cell lines which could not produce CYFRA 21-1, conjectured that some abnormalities in exon 1 or the 5'-region upstream from exon 1. In conclusion, it was demonstrated that the release of CK19 fragment was closely related to the expression of mRNA for CK19, and the possibility that genomic change of CK19 DNA down-regulated the expression of mRNA for CK19 was suggested.

Increased intensity of lung infiltrates at the side of lung cancer in patients with lung cancer associated with pulmonary fibrosis.

It has been reported that lung cancer is frequently associated with idiopathic pulmonary fibrosis (IPF). The purpose of this study was to compare the intensity of lung infiltrates between the side associated with lung cancer and the side without lung cancer. Twenty-three patients (24 lung cancers) with primary lung cancer associated with pulmonary fibrosis were retrospectively evaluated. Chest CT findings were evaluated by three expert radiologists using the intensity scores. In 16 of the 23 patients, it was possible to compare the intensity of lung infiltrates between both sides of the lungs. As a result, increased intensity at the side in which lung cancer developed was demonstrated in 12 of 16 patients (75%). In the remaining four patients, intensity of lung infiltrates was the same in both lungs. In operated patients as well as autopsied patients, it was possible to evaluate the pathological findings of lung tissues around cancer cells. This study clearly demonstrates that the intensity of lung infiltrates increased at the side in which lung cancer developed.

Differential expression of NGFI-B and RNR-1 genes in various tissues and developing brain of the rat: comparative study by quantitative reverse transcription-polymerase chain reaction.

NGFI-B and RNR-1 are closely related transcription factors that constitute a distinct subclass within the steroid/thyroid hormone receptor superfamily. They have been implicated in neuronal differentiation, neuroendocrine regulation of adrenocortical function and T-cell apoptosis. In this study, we measured and compared NGFI-B and RNR-1 mRNA levels in various adult rat tissues and in the developing rat brain by means of the quantitative reverse transcription-polymerase chain reaction. The use of RNA standards synthesized in vitro allowed direct comparison of the amount of the transcripts of these two genes. We demonstrated that the transcripts of both genes were present in all tissues examined although the expression levels widely varied. We found the highest constitutive expression of both genes in the pituitary. High levels of NGFI-B were also expressed in the cerebral cortex, muscle, ventral prostate, thymus and adrenal glands, whereas high levels of RNR-1 expression were restricted to the pituitary and cerebral cortex. These findings were consistent with the notion that NGFI-B and RNR-1 are involved in various signal transduction systems in diverse cell types. The amount of NGFI-B mRNA was greater than that of RNR-1 mRNA in all adult rat tissues, with the highest ratio of NGFI-B relative to RNR-1 expression in the muscle and leukocytes. In contrast, fetal rat brain showed relatively high RNR-1 gene expression. These findings suggested that the NGFI-B and RNR-1 genes are differentially expressed in a tissue-specific and developmentally regulated manner.

Elevation of cytokeratin 19 fragment (CYFRA 21-1) in serum of patients with radiation pneumonitis: possible marker of epithelial cell damage.

Cytokeratin 19 fragment (CYFRA 21-1) level in serum have already been documented as a useful tumor marker for lung cancer. In the present study, we hypothesized that CYFRA 21-1 increases in the sera of patients with radiation pneumonitis, resulting from epithelial cell damage. We measured CYFRA 21-1 in the sera of patients with radiation pneumonitis and evaluated the correlation between CYFRA 21-1 level and severity of radiation pneumonitis as well as clinical course. We studied 16 patients diagnosed with radiation pneumonitis associated with primary lung cancer. CYFRA 21-1 levels in the sera of patients with diffuse radiation pneumonitis (n = 6) significantly increased compared to normal smokers (n = 10) or patients with local radiation pneumonitis (n = 10). CYFRA 21-1 values in sera changed according to the progression or improvement of the diffuse radiation pneumonitis. An immunohistochemical study using pulmonary tissues obtained from autopsied patients with radiation pneumonitis demonstrated that the hyaline membrane and proliferating type II pneumocytes were strongly stained by the anti-human cytokeratin 19 antibody. Our data demonstrated that CYFRA 21-1 was increased in patients with diffuse radiation pneumonitis. Since CYFRA 21-1 is widely used as a tumor marker for lung cancer, this evidence should be noted especially in irradiated patients.

Circulating bronchoepithelial cells expressing mRNA for surfactant protein A in patients with pulmonary fibrosis.

There are several unsolved clinical findings in patients with idiopathic pulmonary fibrosis (IPF); (i) predominance of fibrosis in the lower lung fields, (ii) digital clubbing, and (iii) patchy distribution of pulmonary fibrosis. To explain these unsolved problems, we hypothesized that regenerated or premature bronchoepithelial cells may circulate in the blood in patients with IPF. To prove this, we performed the reverse transcriptase-polymerase chain reaction (RT-PCR) for cytokeratin 19 (CK19) and pulmonary surfactant protein A (SPA) in peripheral blood in patients with IPF and pulmonary fibrosis associated with collagen vascular disorders. In addition, 20 patients with chronic pulmonary emphysema as a disease control and 19 normal volunteers were also evaluated for the existence of circulating bronchoepithelial cells. RT-PCR analysis showed that CK19 was expressed in 12 of 38 blood samples (31.6%) of IPF and pulmonary fibrosis associated with collagen vascular disorders, seven of 20 (35.0%) blood samples of chronic pulmonary emphysema, and four of 19 (21.1%) blood samples of normal volunteers. mRNA for SPA was positive in eight of 38 (21.1%) blood samples of IPF. In contrast, SPA expressing cells were not detected in any blood samples obtained from patients with chronic pulmonary emphysema or normal volunteers. This evidence suggests that there were some circulating bronchoepithelial cells expressing mRNA for SPA in peripheral blood of patients with IPF and pulmonary fibrosis associated with collagen vascular disorders.

The role of anti-epithelial cell antibodies in the pathogenesis of bilateral radiation pneumonitis caused by unilateral thoracic irradiation.

Two cases of bilateral radiation pneumonitis associated with unilateral thoracic irradiation against lung cancer are described. Both patients died of respiratory failure and autopsy was performed. Histologically, bilateral diffuse alveolar damage was demonstrated in both cases, associated with marked organization of hyaline membrane in one case (case 1). In addition, numerous hyperplastic type II pneumocytes which strongly expressed cytokeratins 8, 18 and 19 were observed. In both patients' sera, antibodies against cytokeratin 8, 18 and 19 were demonstrated by a Western immunoblot. The possible association between autoantibodies to cytokeratins and diffuse alveolar damage observed in patients with bilateral radiation pneumonitis are discussed.

Non-specific interstitial pneumonia and Chlamydia pneumoniae infection.

Recently, the clinical features of non-specific interstitial pneumonia (NSIP) have been described. We hypothesize that recurrent infection caused by Chlamydia pneumoniae may play a role in the pathogenesis of NSIP. To prove this, we quantified serum IgA and IgG antibodies against C. pneumoniae using the enzyme linked-immunosorbent assay kit. The study included 15 patients diagnosed with NSIP, 20 patients with chronic obstructive pulmonary diseases (COPD) as disease group, and 27 control subjects. IgA antibody against C. pneumoniae was positive in 12 of 15 patients with NSIP, in 16 of 20 patients with COPD, and in 14 of 27 control subjects. IgG antibody against C. pneumoniae was positive in 14 of 15 patients with NSIP, in 17 of 20 patients with COPD, and in 16 of 27 control subjects. If the cut off value (mean +/- 2SD, index more than 3.0) was introduced, IgA and/or IgG antibodies against C. pneumoniae were positive in 8 of 15 patients with NSIP (53.3%), in 9 of 20 patients with COPD (45%), and in 2 of 27 control subjects (7.4%). These results suggest that infection of C. pneumoniae might play a role in the pathogenesis of NSIP.

A salivary gland-type monomorphic adenoma with trabecular proliferation in the lung.

Salivary gland-type adenoma, especially monomorphic adenoma in the lung is very rare. We report a 66-year-old previously healthy woman who developed cough, hemosputum, and fever. Bronchoscopy revealed an endobronchial mass in the proximal portion of the left lower lobe bronchus. Following left lower lobectomy, the pathological diagnosis was an unusual monomorphic adenoma of the salivary gland-type with trabecular proliferation.

Clinical features of three fatal cases of non-specific interstitial pneumonia.

We describe the clinical courses of the 3 fatal patients (2 females and 1 male) with idiopathic non-specific interstitial pneumonia (NSIP) among 24 patients with NSIP. Lung biopsies were diagnosed to be NSIP group II in all patients. The clinical courses from onset to death of these 3 patients were 41 months, 46 months, and 91 months. A follow-up chest CT demonstrated no apparent honey-comb formation. We found that i) about 20% of patients with NSIP died of respiratory failure, ii) in the chest CT findings, apparent honey-comb formation was rare even just before death, iii) prediction of the prognosis based on the histological findings was difficult. This is the first report to describe the clinical features of deceased patients with idiopathic NSIP; the incidence of fatal cases was considered to range from 10 to 20%.

Thymic carcinoma associated with a high serum level of interleukin 6 diagnosed through the evaluation for asymptomatic elevation of acute-phase reactants.

A case of thymic squamous cell carcinoma producing interleukin-6 (IL-6) is reported. C-reactive protein (CRP), white blood cell (WBC) count, and serum IL-6 had been elevated, and normalized immediately after tumorectomy. IL-6 in the culture supernatant from the tumor was significantly elevated and the expression of IL-6 mRNA was demonstrated in the tumor by a reverse transcriptase polymerase chain reaction method. Immunohistochemical study demonstrated that tumor cells were stained positive with an anti-IL-6 antibody. IL-6 derived from the tumor cells reflected the increase CRP and WBC counts. This case suggested that asymptomatic elevation of acute-phase reactants might be clues for the diagnosis of an IL-6 producing tumor.