Chiharu Yoshii

Severe Pneumonia with Leptotrichia sp. Detected Predominantly in Bronchoalveolar Lavage Fluid by Use of 16S rRNA Gene Sequencing Analysis

ABSTRACT We present the first case of severe pneumonia possibly caused by Leptotrichia species with oral bacteria. This was found in a healthy but elderly subject whose bronchoalveolar lavage fluid was analyzed by 16S rRNA gene sequencing analysis. The combination of this method and microscopic observation provided useful information for diagnosis and treatment.

Detection of MALT1 Gene Rearrangements in BAL Fluid Cells for the Diagnosis of Pulmonary Mucosa-Associated Lymphoid Tissue Lymphoma

BACKGROUND Mucosa-associated lymphoid tissue (MALT) lymphoma constitutes approximately 90% of primary pulmonary lymphoma, and the diagnosis of pulmonary MALT lymphoma often requires invasive methods such as surgical lung biopsy. Chromosomal rearrangements involving MALT lymphoma translocation gene 1 (MALT1) have been reported to be specific for MALT lymphoma. The combination of BAL and cytologic approaches with molecular methods is useful for the diagnosis of lymphoproliferative disorders. Therefore, we examined the detection of MALT1 gene rearrangements in BAL fluid (BALF) cells for the diagnosis of MALT lymphoma. METHODS We determined the percentage of BALF cells with MALT1 gene rearrangements by using the fluorescence in situ hybridization (FISH) method in 10 patients suspected to have pulmonary MALT lymphoma. RESULTS MALT1 gene rearrangements in BALF cells were found in four of five cases with pulmonary MALT lymphoma (percentage of BALF cells with MALT1 gene rearrangements: 21.8% ± 6.8%). On the other hand, MALT1 gene rearrangements in BALF cells were negative in the five cases without pulmonary MALT lymphoma and one case with pulmonary MALT lymphoma. CONCLUSION These results suggest that the detection of MALT1 gene rearrangements in BALF cells is useful for the diagnosis of pulmonary MALT lymphoma, as it is a specific method that is less invasive than surgical biopsy. Because of the small number of patients in this study, further investigations are necessary to evaluate the detection rate of MALT1 gene rearrangements in BALF cells from patients with pulmonary MALT lymphoma.

Assessment of Pathologically Diagnosed Patients with Castleman’s Disease associated with Diffuse Parenchymal Lung Involvement Using the Diagnostic Criteria for IgG4-Related Disease

BackgroundIgG4-related disease (IgG4RD) is a recently recognized disease entity. Differentiating IgG4RD from plasma cell type Castleman’s disease (PCD) is important but also difficult using only pathological findings. In addition, little is known about the association between these two diseases with diffuse parenchymal lung involvement.MethodsWe analyzed the serum IgG4 levels and the ratio of IgG4/IgG-positive plasmacytes in the lung and lymph node specimens of eight patients previously pathologically diagnosed of PCD with diffuse parenchymal lung involvement (DL-PCD). We also compared the clinical and laboratory findings observed in these patients.ResultsSix of the eight patients exhibited abundant IgG4-positive plasmacytes in the lung and lymph node tissues and elevated serum IgG4 levels, thereby fulfilling the diagnostic criteria of IgG4RD with DL (DL-IgG4RD) in addition to having obstructive phlebitis and massive lymphoplasmacytic infiltration with fibrosis. However, three of these six patients exhibited higher levels of serum interleukin-6 and were still diagnosed with DL-PCD. Accordingly, three of these eight patients were considered as IgG4RD with DL (DL-IgG4RD), and the other five patients were ultimately given a diagnosis of DL-PCD. These two diseases have different characteristics in terms of age, symptoms, serum levels of C-reactive protein, and IgA, complicating allergic disorders, response to corticosteroids, and prognosis.ConclusionsThis is the first report to show a high prevalence of DL-IgG4RD in DL-PCD patients, although additional large investigations are necessary. Clinical and laboratory findings are important for distinguishing between these two diseases in other organs, as previously described.

Calcitonin gene-related peptide stimulates proliferation of alveolar epithelial cells.

BackgroundAlveolar epithelial cells are known as progenitor cells for the restoration from the damage in the lung. Calcitonin gene-related peptide (CGRP) has been reported to play an important role in the proliferation of various types of epithelial and endothelial cells. We investigated the effects of CGRP on the proliferation of alveolar epithelial cells in vitro and in vivo.MethodsA549 cells were cultured in Dulbecco Modified Eagle Medium with 5% fatal bovin serum for 24 hours, then CGRP was added in vitro. The proliferation of DNA synthesis was measured using 5-bromo-2-deoxyuridine, an analog of thymidine, by enzyme-linked immunosorbent assay.As one intracellular response to CGRP, we examined activation of p44/42- extracellular signal-regulated kinase (ERK) pathway by adding CGRP, using western blotting method.Recombinant adenovirus encoding nuclear-targeted-human β-CGRP (rhCGRP) was administered into Male Wister rat (n = 5, 10 weeks old) lungs by intratracheal instillation in vivo. 7 days after the administration of CGRP, rat lungs were harvested and histological findings and immunohistochemical staining of proliferating cell nuclear antigen (PCNA) were evaluated to examine cell proliferation.ResultsIn vitro study, CGRP increased the proliferation of A549 cells in a dose and time dependent manner. CGRP8-37 (inhibitor of CGRP receptor) decreased CGRP induced proliferation of DNA synthesis. Phosphorylation of ERK pathway was observed within 15 minutes and peaked in one hour. U0126 (inhibitor of ERK pathway) decreased CGRP induced proliferation of DNA synthesis.In vivo study, histological examination of the lung indicated proliferation of alveolar epithelial cells in the rhCGRP-treated group and the nuclei of alveolar epithelial cells were positive for PCNA immunostaining.ConclusionIn this study, we conclude that CGRP stimulates proliferation of human alveolar epithelial cells in vivo and in vitro.

Varenicline does not increase serum BDNF levels in patients with nicotine dependence.

Varenicline, α4β2 nicotinic acetylcholine receptor (nAChR) partial agonist, is a new class of medications for treating nicotine dependence. As an α4β2 nAChR partial agonist, varenicline serves to reduce nicotine withdrawal symptoms, while high‐affinity binding of the agonist mitigates the reinforcing effects of smoking. In the present study, we compared serum brain‐derived neurotrophic factor (BDNF) levels of nicotine dependence and nonsmokers, and we investigated changes in serum BDNF levels after 8 weeks of treatment with varenicline. Patients met the DSM‐IV criteria for nicotine dependence. Both the Fagerström test for nicotine dependence (FTND) and the Tobacco Dependence Screener (TDS) were used. Serum BDNF levels and breath carbon monoxide (CO) levels were measured before and 8 weeks after varenicline treatment. Fourteen of 16 subjects (87.5%) stopped smoking within 12 weeks of varenicline treatment. Thirteen healthy nonsmokers who never had previously smoked were randomly selected as a control group. Serum BDNF levels of patients before treatment (4.8 ± 3.8 ng/ml) were significantly lower than those in the control group (12.4 ± 6.13 ng/ml). Serum BDNF levels had not increased from baseline (4.8 ± 3.8 ng/ml) to 8 weeks after varenicline treatment (3.0 ± 1.1 ng/ml) of patients. These results suggest that smoking might decrease serum BDNF levels and that treatment with varenicline for 8 weeks, combined with 12 weeks of not smoking, does not increase serum BDNF levels in smokers. Copyright

Osteosclerotic lesions in patients treated with gefitinib for lung adenocarcinomas: a sign of favorable therapeutic response

ObjectiveTo assess the frequency of osteosclerotic changes on CT that appeared after treatment with gefitinib in patients with lung adenocarcinoma and the relationship between the osteosclerotic changes and the response to the therapy.Materials and methodsOur study included 41 patients with lung adenocarcinoma who underwent chest CT both before (CTpre) and after (CTpost) starting treatment with gefitinib. The presence or absence of bone metastases was assessed on the CTpre, and the interval bony change after the therapy was classified as lytic, sclerotic, or no changes on the CTpost. The relationship between treatment results of primary lung cancer and interval bony changes was evaluated.ResultsOsteosclerotic lesions were identified in 11 patients (27%) on CTpost; in 6 of 11 patients osteosclerotic lesions newly appeared where the CTpre showed no bone metastasis before the gefitinib therapy. There were significant differences in the therapeutic response of the primary cancers (P < 0.001) and in the survival rate (P < 0.01) in patients with osteosclerotic changes versus those without osteosclerotic changes.ConclusionOsteosclerotic changes on CT, observed after gefitinib treatment in patients with lung adenocarcinomas, may be an indicator of a good therapeutic response.

Features of idiopathic pulmonary fibrosis with organizing pneumonia.

To characterize the clinical features of patients with idiopathic pulmonary fibrosis (IPF) having organizing pneumonia (OP), we retrospectively reviewed the clinical charts, chest X-rays, CT scans, and transbronchial lung biopsy (TBLB) specimens of patients with IPF. Patients with IPF and OP had a subacute onset of symptoms (within 2 months) (87.5%), leukocytosis (> 10,000/mm3) (62.5%), and a strong C-reactive protein (CRP) reaction (> 3+) (75%). Some of these features were distinctly different from those of IPF patients without OP (subacute onset of symptoms 0%, leukocytosis 0%, strong CRP reaction 16.7%). In the patients with IPF with OP, A-aDo2 and semiquantitative scores of chest X-ray abnormalities improved significantly after prednisolone treatment. Those abnormalities improved only slightly in the patients with IPF without OP. Diffusing capacity remained decreased and abnormal interstitial infiltration persisted, even after prednisolone treatment in the patients with IPF with or without OP. Clinical features of IPF patients with OP differed from those of patients with IPF without OP. IPF patients with OP showed good clinical response to corticosteroid therapy. These findings warrant further study of the presence of OP in TBLB specimens in predicting corticosteroid responsiveness and prognosis of patients with IPF.

Relationship between inflammatory cells in bronchoalveolar lavage fluid and pathologic changes in the lung interstitium

Bronchoalveolar lavage (BAL) is recognized as an important research tool for various lung diseases, but it is still uncertain whether inflammatory cells in BAL fluid (BALF) accurately reflect pathologic changes in the lung interstitium. We used a morphometric method to quantify the density of inflammatory cells in the lung interstitium by utilizing a computer-aided graphic analyzer and compared those findings with BALF results. Two types of animal models were studied, i.e., endotoxemia (Escherichia coli endotoxin) and hypersensitivity pneumonitis (inhaled ovalbumin). Male Wistar rats were used; the right lungs were lavaged and the left lungs were prepared for morphometric study. In the endotoxemia model, the neutrophil fraction in BALF and the neutrophil density in the lung interstitium correlated significantly at 18 h (r = 0.81, p < 0.05) and 24 h (r = 0.81, p < 0.05) but not at any other time points after injection. In the hypersensitivity pneumonitis model, the neutrophil fraction in BALF and the neutrophil density in the lung interstitium correlated significantly (r = 0.80, p < 0.05) only at 3 h after inhalation. The lymphocyte fraction in BALF and the lymphocyte density in the lung interstitium were correlated positively at 3 h (r = 0.83, p < 0.05), 1 day (r = 0.82, p < 0.05), 2 days (r = 0.67, p = NS), and 4 days (r = 0.87, p < 0.05), but not at 6 days after inhalation. Our data suggest that neutrophil fraction in BALF does not reflect neutrophil populations in the lung interstitium except at the time of maximal neutrophil count in lung lavage. For lymphocytes in the hypersensitivity pneumonitis model, those in BALF and in the lung interstitium roughly correlate in the majority of measurements.

Evaluation of a rapid immunochromatographic ODK0501 assay for detecting Streptococcus pneumoniae antigens in the sputum of pneumonia patients with positive S. pneumoniae urinary antigens.

BACKGROUND A novel, rapid and noninvasive test (ODK0501, RAPIRUN(®)Streptococcus pneumoniae) uses polyclonal antibodies to detect C polysaccharide of S. pneumoniae derived from sputum samples using an immunochromatographic assay. We evaluated its usefulness in Japanese patients with pneumonia who exhibited positive urinary antigen tests for S. pneumoniae (BinaxNOW(®)S. pneumoniae). PATIENTS AND METHODS Forty adult patients with pneumonia treated between May 2011 and August 2013 were enrolled. Bacterial cultures, Gram staining and ODK0501 assays of sputum as well as urinary antigen tests for S. pneumoniae using urine samples obtained from the same patients were performed upon admission, the fourth day after starting antimicrobial treatment and at the end of the antimicrobial treatment. RESULTS Twenty-seven of the 40 patients were positive for ODK0501, while a negative result for ODK0501 was associated with low-quality sputum samples according to the Geckler classification of sputum. The sensitivity and specificity of the ODK0501 assay in the 40 patients were 90.9% and 61.1%, respectively, based on the culture results. The results obtained with this kit were more favorable than those observed on Gram staining. The ODK0501 assay also showed a rapid reaction to the disappearance of S. pneumoniae in the sputum samples, while approximately 80% of the patients exhibited persistent positive results on the urinary antigen detection tests at the end of treatment. CONCLUSIONS The ODK0501 test is a noninvasive, rapid and accurate tool for diagnosing respiratory infections caused by S. pneumoniae, although good quality sputum must be obtained prior to adequate treatment with antibiotics.

Pathology and Mechanism of Lung Toxicity Following Inhalation of Hair Spray in Rats

In order to elucidate the pathology and mechanism of lung toxicity induced by chronic hair spray inhalation, male Wister rats 9 wk of age were exposed to a uniform concentration of hair spray for up to 12 wk using a jet nebulizer. The aerosol concentration to which the rats were exposed was about 7 g/m3. Differential cell counts in bronchoalveolar lavage fluid (BALF), the expression of several cytokines from alveolar macrophage using the reverse-transcription polymerase chain reaction (RT-PCR) method, and histopathologic evaluation using a computeraided graphic analyzer (IBAS) were conducted 1, 4, 8, and 12wkafter exposure.Over the passage of time, neutrophils and macrophages increased in BALF, and neutrophils infiltrated in the lung interstitium from the peribronchial interstitium to the alveolar septum. Alveolar macrophages showed increased expression of both the mRNA of tumor necrosis factor (TNF)-α and the mRNA of the chemokines macrophage inflammatory protein 2 (MIP2) and cytokine-induced neutrophil attractant (CINC). Fromthese findings, chronic inhalation of hair spray is considered to induce at first intra-alveolar accumulation and activation of alveolar macrophages, followed by recruitment of neutrophils in the lung through the expression of proinflammatory cytokine, CINC, and MIP2, which cause predominantly neutrophilic inflammation in the lung.