GLCCI1 variant accelerates pulmonary function decline in patients with asthma receiving inhaled corticosteroids.

BACKGROUND: In steroid-naive patients with asthma, several gene variants are associated with a short-term response to inhaled corticosteroid (ICS) treatment; this has mostly been observed in Caucasians. However, not many studies have been conducted for other ethnicities. Here, we aimed to determine the relationship between the annual decline in forced expiratory flow volume in one second (FEV1 ) and the variant of the glucocorticoid-induced transcript 1 gene (GLCCI1) in Japanese patients with asthma receiving long-term ICS treatment, taking into account the effect of high serum periostin levels, a known association factor of pulmonary function decline and a marker of refractory eosinophilic/Th2 inflammation. METHODS: In this study, 224 patients with asthma receiving ICS treatment for at least 4 years were enrolled. The effects of single-nucleotide polymorphisms (SNPs) in GLCCI1, stress-induced phosphoprotein 1 (STIP1), and T gene on the decline in FEV1 of 30 ml/year or greater were determined. RESULTS: Besides the known contributing factors, that is, the most intensive treatment step, ex-smoking, and high serum periostin levels (≥95 ng/ml), the GG genotype of GLCCI1 rs37973, and not other SNPs, was independently associated with a decline in FEV1 of 30 ml/year or greater. When patients were stratified according to their serum periostin levels, the GG genotype of rs37973 was significantly associated with blood eosinophilia (≥250/µl) in the high serum periostin group. CONCLUSIONS: A GLCCI1 variant is a risk factor of pulmonary function decline in Japanese patients with asthma receiving long-term ICS treatment. Thus, GLCCI1 may be associated with response to ICS across ethnicities.

Non-genomic inhibitory effect of glucocorticoids on activated peripheral blood basophils through suppression of lipid raft formation.

We investigated the non-genomic effects of glucocorticoids (GCs) on inhibition of plasma membrane lipid raft formation in activated human basophils. Human basophils obtained from house dust mite (HDM)-sensitive volunteers were pretreated with hydrocortisone (CORT) or dexamethasone (Dex) for 30 min and then primed with phorbol 12-myristate 13-acetate (PMA, 10 ng/ml) or HDM (10 µg/ml). The expression of CD63, a basophil activation marker, was assessed by flow cytometry. Membrane-bound GC receptors (mGCRs) were analysed by flow cytometry and confocal laser microscopy. Lipid rafts were assessed using a GM1 ganglioside probe and visualization by confocal laser microscopy. Pretreatment of basophils with CORT (10(-4) M and 10(-5) M) and Dex (10(-7) M) significantly inhibited CD63 expression 20 min after addition of PMA or HDM. The inhibitory effects of GCs were not altered by the nuclear GC receptor (GCR) antagonist RU486 (10(-5) M) or the protein synthesis inhibitor cycloheximide (10(-4) M) (P < 0·05). CORT coupled to bovine serum albumin (BSA-CORT) mimicked the rapid inhibitory effects of CORT, suggesting the involvement of mGCRs. mGCRs were detectable on the plasma membrane of resting basophils and formed nanoclusters following treatment with PMA or HDM. Pretreatment of cells with BSA-CORT inhibited the expression of mGCRs and nanoclustering of ganglioside GM1 in lipid rafts. The study provides evidence that non-genomic mechanisms are involved in the rapid inhibitory effect of GCs on the formation of lipid raft nanoclusters, through binding to mGCRs on the plasma membrane of activated basophils.

Health-related quality of life does not predict mortality in idiopathic pulmonary fibrosis.

BACKGROUND: Although health-related quality of life (HRQL) has recently been considered to be an important outcome in clinical trials of idiopathic pulmonary fibrosis (IPF), its relationship with survival is unknown. OBJECTIVE: To determine the prognostic significance of HRQL scores in IPF assessed with the SGRQ. DESIGN: Eighty-seven consecutive patients with IPF, who had undergone evaluations and completed the St. George's Respiratory Questionnaire (SGRQ) at diagnosis were included in this study, as is the general practice. Cox proportional hazards analyses were performed to examine the relationship between HRQL scores and survival. RESULTS: The mean observation period was 44.2 +/- 29.6 mo, in the course of which 54 patients (62.0%) died. Univariate analysis revealed that the activity scores in the SGRQ(HR: 1.016, 95% CI: 1.004-1.029, P = 0.01) were significantly predictive of survival, although the symptoms, impacts, and total scores were not significantly related to mortality from all causes. However, multivariate analysis revealed that only the forced vital capacity percent predicted was a significant predictor of survival, and that the activity score in the SGRQwas not significantly related to mortality. CONCLUSIONS: There was no significant relationship between HRQL evaluated with the SGRQ and the subsequent mortality in IPF. The present negative result might suggest that HRQL is measuring an aspect other than one from physiological and functional impairment or disability.

Histopathology of the airway epithelium in an experimental dual-phase model of bronchial asthma.

BACKGROUND: Lesions of trachea cuticles are a pathological histological characteristic of bronchial asthma. Furthermore, collected tracheal cuticles desquamated from the respiratory tract are found in patients' sputum when asthma attacks occur or after the induction of allergen inhalation. From these facts, it is assumed that desquamation of trachea cuticle cells is a pathological symptom of bronchial asthma. However, there has not been any chronological report of desquamation of trachea cuticles through the process of bronchial asthma attacks. OBJECTIVE: For this report, we made an experimental bronchial asthma model using guinea pigs, and conducted chronological examinations of trachea cuticle lesions related to pathological symptoms of bronchial asthma using a transmission electron microscope and a scanning electron microscope. METHODS: The experimental asthma models were made by injection of ovalbumin into the abdominal cavity of guinea pigs. Then the airway responses to inhaled aerosolized ovalbumin were induced. The trachea were enucleated and examined under an optical microscope, a transmission electron microscope (hereafter abbreviated as TEM), and a scanning electron microscope (hereafter abbreviated as SEM) after 1, 2, 4, 8, 12, 24 h and 7 days after the immediate airway responses. RESULTS: Intercellular oedema of ciliated epithelium was observed in the sensitization groups immediately after the immediate airway response. SEM observation revealed increased mucus secretion and shortening of cilium. A slight case of desquamation or deciduation of ciliated epithelium was also beginning to appear. TEM observation revealed a dilation of ciliated epithelium intervals. Infiltration of eosinophilic leucocytes was already detectable beneath the ciliated epithelium. The degree of ciliated epithelium desquamation and infiltration of eosinophilic leucocytes progressed with time. When the late airway response occurred 4 hours later, eosinophilic leucocytes had increased drastically, and ciliate epithelium had desquamated to the extent that basal cells were exposed. Seven days after the immediate airway response, epithelium intercellular oedema had improved, and cilium had been reproduced. CONCLUSION: These results suggest that desquamation of epithelium caused by trachea cuticle lesions appears at an early stage of an asthma attack, owing to the contraction of the trachea, and that the damage is intensified by the infiltration of eosinophilic leucocytes.

[A case of Sjögren's syndrome with pleural effusion].

A 45-year-old woman was admitted to our hospital because of a fever. A round erythema was noted on the skin, suggesting collagen disease. Bilateral pleural effusion developed during hospitalization, and serum and pleural effusion were positive for antinuclear antibody, RA factor, anti-SS-A antibody, and anti-SS-B antibody. A diagnosis of Sjögren's syndrome was made on the basis of reduced lacrimation and the histological findings in a biopsy specimen from the lip. The cells in the pleural effusion were predominantly lymphocytes, and so a pleural lesion associated with Sjögren's syndrome was suspected, but reports of this condition have been scarce. Good therapeutic results were obtained by corticosteroid administration. Sjögren's syndrome should be considered in the differential diagnosis of pleural effusion associated with collagen disease.

Effect of anti-ICAM-1 on bronchial response: bronchoalveolar lavage fluid (BALF) and ultrastructural changes of bronchial epithelium in guinea pigs with dual phase bronchial response.

Eosinophils play an important role in the development of bronchial asthma, and the association between ICAM-1 and activation and migration of local eosinophils is attracting attention. Using an asthmatic model of dual phase bronchial response, the effects of anti-ICAM-1 antibody on the airway resistance, cell composition in the bronchoalveolar lavage fluid (BALF) and ultrastructure of bronchial ciliated epithelium were examined under the provoked response by inhalation of the antigen. By administration of anti-ICAM-1 antibody, the late asthmatic response (LAR) was suppressed. In the examination of bronchoalveolar lavage fluid, a significant decrease in eosinophils was found in LAR. In examining transmission and scanning electron microscopies, no difference was found in the immediate asthmatic response, but marked suppression of deciduation of bronchial ciliated epithelium was observed in LAR. These results indicated that anti-ICAM-1 antibody suppressed bronchial asthmatic attack, mainly in LAR, by controlling differentiation and migration of eosinophils.

Effects of ONO-1078 (pranlukast) on cytokine production in peripheral blood mononuclear cells of patients with bronchial asthma.

BACKGROUND: ONO-1078 (pranlukast) is a leukotriene receptor antagonist developed in Japan. This drug has been shown to be useful in oral treatment of bronchial asthma. The present study was undertaken to assess the effects of this drug on the production of cytokines in the peripheral blood mononuclear cells of patients with asthma under stimulation with specific antigens. METHODS: Peripheral blood mononuclear cells from mite antigen-positive asthmatic patients (immunoglobulin E RAST score > 3) were incubated for 72 h in the presence of mite antigen (10 microg/mL). The supernatant of the culture was subjected to enzyme-linked immunosorbent assay (ELISA) to quantify interleukin (IL) -4, IL-3, IL-5, and granulocyte macrophage-colony stimulating factor (GM-CSF). Other peripheral blood mononuclear cells from the same patients were incubated for 72 h in the presence of both mite antigen (10 microg/mL) and ONO-1078 (0.5, 1, or 10 microg/mL), followed by ELISA of the supernatant to quantify the cytokines. RESULTS: Production of IL-4, IL-5, and GM-CSF by mononuclear cells under stimulation with mite antigen was markedly suppressed when they were exposed to ONO-1078 at a concentration of 10 microg/mL. CONCLUSION: The results suggest that ONO-1078 acts directly on peripheral blood mononuclear cells and that blockade of leukotriene receptors on blood mononuclear cells by the cysteinyl-leukotriene receptor antagonist (LTRA) pranlukast (ONO-1078) can dose-dependently inhibit release of immunoreactive TH2-type cytokines (IL-3, IL-4, GM-CSF, and possibly IL-5), but not of the TH1-type cytokine IL-2, when stimulated by mite allergen in vitro. The data may provide clues to the mechanism by which a number of LTRA including zafirulukast and montelukast can reduce airway, sputum and blood eosinophil counts in clinical asthma. It supports animal studies showing that anti-IL-5 antibodies partially block cys-LT-induced airway eosinophilia, suggesting that cys-LTs may cause secondary release of IL-5 from an unknown cell-type. These findings indicate that ONO-1078 suppresses the production of IL-4 (a cytokine that affects IgG antibody production), IL-5, and GM-CSF (cytokines that affect eosinophil activation) by peripheral blood mononuclear cells under stimulation with specific antigens in patients with bronchial asthma. Because of its anti-inflammatory effects, ONO-1078 should be useful in the treatment of bronchial asthma.

Effects of saiboku-to on the survival of human eosinophils.

In recent years, bronchial asthma has come to be regarded as a chronic inflammatory disease of the respiratory tract, with mast cells, lymphocytes and eosinophils playing important roles in its pathogenesis. Proteins contained in eosinophil granules, especially major basic protein (MBP), eosinophil cationic protein (ECP), eosinophil-derived neurotoxin (EDN) and eosinophil peroxidase (EPO), can cause tissue injury. When stimulated, eosinophils release mediators such as leukotriene C4 (LTC4) and platelet activating factors (PAF). Thus, they are recognized as effector cells that are actively involved in the development of allergic inflammation. In this study, eosinophils from healthy volunteers were used to investigate the effects of Saiboku-to on eosinophils whose survival had been prolonged through stimulation with eosinophil-activating cytokines such as interleukin (IL)-3, IL-5 and granulocyte macrophage colony stimulating factors (GM-CSF). As a result, the cytokine-enhanced survival of eosinophils was significantly shortened by the addition of Saiboku-to. These findings suggest that Saiboku-to has the potential to inhibit allergic responses by directly affecting eosinophils which are related to allergic inflammation.

Intrapleural administration of cisplatin and etoposide to treat malignant pleural effusions in patients with non-small cell lung cancer.

BACKGROUND: To determine the efficacy, toxicity and pharmacokinetics of intrapleural cisplatin (CDDP) and etoposide as a treatment for malignant pleural effusions (MPE) in patients with non-small cell lung cancer (NSCLC). METHODS: Seventy patients with MPE associated with NSCLC were enrolled in this study. In 68 patients, a catheter was inserted into the pleural cavity, within 24 h after complete drainage of the pleural effusion, CDDP (80 mg/m2) and etoposide (80 mg/m2) were simultaneously administered successfully via the catheter and the catheter was clamped. Seventy-two hours later, the catheter was unclamped to allow drainage. The catheter was removed when the accumulated intrapleural fluid decreased to 20 ml or less per day. RESULTS: The pharmacokinetic profiles showed high maximum concentrations of CDDP (free form, 88 microg/ml) and etoposide (182. 4 microg/ml) in intrapleural fluids. CDDP did not remain for a long period (free form, beta-phase half-life = 10.51 h) in the fluids, while etoposide persisted for a long period (beta-phase half-life = 62.53 h). The overall response rate was 46.2%, the median survival time 32.3 weeks, the 1-year survival rate 28.7% and the 2-year survival rate 12.8%. The most serious adverse reactions were WHO grade 3 anemia (3 patients), grade 3 nausea and vomiting (17 patients), grade 3 constipation (1 patient), grade 3 pulmonary toxicity (1 patient), grade 4 fever (1 patient), grade 3 infection (1 patient) and grade 3 mental disorder (1 patient). CONCLUSION: Intrapleural administration of CDDP and etoposide was an effective and acceptable regimen for patients with MPE due to NSCLC.

Effects of theophylline on lymphocyte phosphodiesterase activity.

1. To clarify part of the bronchodilating effects of theophylline, lymphocyte cyclic AMP (cAMP) and phosphodiesterase (PDE) activity in asthmatic patients and healthy adults were measured. 2. No clear differences in lymphocyte PDE activity were found regarding sexes, ages or the types of asthma. 3. Lymphocyte PDE activity in the asthma group was significantly higher than that in the control group. 4. Lymphocyte cAMP significantly increased at 30 and 60 min after intravenous injection of aminophylline in the asthma group. 5. Lymphocyte PDE activity significantly decreased 60 min after intravenous injection of aminophylline in the asthma group.

Theophylline suppresses the release of interleukin-4 by peripheral blood mononuclear cells.

To elucidate the mechanism of the anti-inflammatory effect of theophylline, peripheral blood mononuclear cells (PBMCs) collected from patients with bronchial asthma who had a RAST score for Dermatophagoides farinae (Df) > = 3 were stimulated with Df antigen, and the interleukin-4 (IL-4) production by the stimulated PBMCs was determined in the presence and absence of theophylline. The stimulated IL-4 production in the PBMCs was significantly suppressed by simultaneous treatment with theophylline (8 microg/ml). Furthermore, the intracellular 3',5'-adenosine monophosphate (cAMP) level following stimulation with Df was significantly higher in PBMCs simultaneously treated with theophylline (8 microg/ml) than in nontreated controls. These results suggest that theophylline suppresses the IL-4 production in inflammatory cells by increasing the intracellular cAMP level; this effect may thus provide an additional basis for the benefit of theophylline in antiasthmatic therapy.

Evaluation of a cytokine combination including thrombopoietin for improved transduction of a retroviral gene into G-CSF-mobilized CD34+ human blood cells.

We examined cell culture conditions with various combinations of cytokines including thrombopoietin (TPO) to obtain the most efficient transduction of recombinant retrovirus vectors into G-CSF-mobilized blood CD34+ cells which were obtained from children and purified with an Isolex 50 system (Baxter; Deerfield, IL). Three different 4-day culture conditions for the stimulation of CD34+ cells were compared in terms of a cell-cycle analysis by fluorometry and gene transduction efficiency as determined by resistance to G418 and NeoR polymerase chain reaction (PCR) for individual colony-forming unit-granulocyte/macrophage (CFU-GM) grown in a methylcellulose culture system. The cytokines tested were: A) interleukin (IL)-6 + stem cell factor (SCF); B) IL-3 + IL-6 + SCF, and C) IL-3 + IL-6 + SCF + TPO. Without a cell culture, the percentage of CD34+ cells in the cell cycle (the percentage of cells in phases S and G2/M) was 4.6%. After a four-day culture (n = 5), this value increased with the addition of IL-3 (22%) or IL-3 + TPO (27%, p < 0.05) as compared to that with the baseline cocktail of IL-6 + SCF (15%). The cell number uniformly increased approximately 10-fold in each culture condition. The average efficiency of gene transfer into incubated CD34+ cells with the corresponding combinations of cytokines was, respectively, 57%, 47%, and 30% for G418-screened CFU-GM and 72%, 68%, and 51% for polymerase chain reaction-positive CFU-GM. A statistically significant difference (p < 0.01) was found for G418/CFU-GM with IL-3 + IL-6 + SCF (57%) versus IL-3 + IL-6 + SCF + TPO (30%). Hence, it is likely that the increased cell proliferation produced by the addition of TPO was not necessarily translated into an increased rate of retroviral-mediated gene transduction, possibly because TPO preferentially induced the differentiation of stem cells into mature progenitors in these culture systems.

[Clinical features of eight cases of opportunistic fungal pneumonias].

We evaluated eight cases of pulmonary mycosis in immuno compromised hosts. The underlying diseases were lung cancer with chemotherapy in one case, post bone marrow transplantation (post BMT) in two cases, acquired immunodeficiency syndrome (AIDS) in one case and bronchial asthma with massive steroid therapy in four cases. The causative fungi were Candida sp. in three cases, Aspergillus sp. in four cases, Tricosporon sp. in one case. Prognosis was guarded despite antifungal treatment. Five cases deteriorated and died of fungal infection. In five cases, who died of deterioration, 31.6 days was required from appearance of abnormal infiltration in the chest X-ray to determination of the causative fungi (including two cases who were diagnosed by autopsy) on the average. In three successfully treated cases, the average duration from the appearance of abnormal infiltration in the chest X-ray for the determination of the causative fungi was 8.3 days. On the contrary, the average duration between the appearance of abnormal infiltration in the chest X-ray and the initiation of antifungal treatment was 2.6 days who died of deterioration and 8.3 days who survived. We conclude that early identification of causative fungi and not quick institution of antifungal treatment was mandatory in the treatment of opportunistic fungal pneumonia.

Effects of cytokines on oxygen radical production by peripheral blood monocytes and alveolar macrophages in patients with lung cancer.

The effects of cytokines (interleukin-2, tumor necrosis factor-alpha and interferon-gamma) on the ability of peripheral blood monocytes and alveolar macrophages to produce oxygen radicals were examined by the chemiluminescence assay in patients with lung cancer. Oxygen radical production by peripheral blood monocytes before stimulation with cytokines was lower in the lung cancer group than in healthy controls, suggesting reduced immune function in lung cancer patients. However, the activity in the lung cancer group was elevated to the control level when the monocytes were stimulated by any of the three aforementioned cytokines. Oxygen radical production by alveolar macrophages did not differ significantly between nonstimulated monocytes from lung cancer patients and those from healthy controls. In the lung cancer group, stimulation of the macrophages with any of the three cytokines elevated their ability to produce oxygen radicals to the same extent as in the control group. The results suggest that stimulation of macrophages by interleukin-2, tumor necrosis factor-alpha or interferon-gamma can exert an antitumor action in patients with lung cancer.

Assessment of serum CYFRA 21-1 in lung cancer.

BACKGROUND: Cytokeratins are the intermediate filaments of the cytoskeletal protein located in normal epithelia, tumor, and cultured cells. Recently, a fragment of cytokeratin subunit 19, referred to as CYFRA 21-1, detected in the serum of patients with nonsmall cell lung cancer, has been reported as a new tumor marker. This article reports the results of a study of serum fragment CYFRA 21-1, measured by immunoradiometric assay, as a marker of lung cancer. METHODS: One hundred fourteen patients with primary lung cancer, 6 patients with malignant solid tumor, 116 patients with a variety of benign diseases, and 29 normal individuals were entered into the study. Serum CYFRA 21-1 levels were obtained by means of immunoradiometric assay using the CYFRA 21-1 EIA (enzyme immunoassay) kit. In addition, we studied other tumor markers, including carcinoembryonic antigen (CEA), squamous cell carcinoma antigen (SCC), and neuron specific enolase (NSE), as a means of lung cancer diagnosis. RESULTS: The diagnostic accuracy and sensitivity of serum CYFRA 21-1 for the detection of lung cancer were highest among the four markers. The serum CYFRA 21-1 levels were most highly elevated in lung carcinoma patients (in particular UICC Stage IV patients) across different histologic types and attained 85.1% sensitivity when using a threshold of 3.5 ng/mL. The diagnostic sensitivity for detecting lung carcinoma was substantially enhanced by means of combined assays of CYFRA 21-1 with CEA overall for lung cancer, with SCC for squamous cell carcinoma, and with CEA for adenocarcinoma. CONCLUSIONS: These findings suggest that serum assays of CYFRA 21-1 are clinically useful for the diagnosis of lung carcinoma.

[Persistent reactive airway dysfunction syndrome after exposure to chromate].

A 42-year-old man was admitted to the hospital because of wheezing and dyspnea that began 20 minutes after accidental exposure to irritant gas containing chromate at a chrome pellet manufacturing plant. The patient had a positive skin reaction to chromite (Cr2O3). Reduction of FEV1.0 was observed 120 minutes after Cr2O3 inhalation challenge. By 150 minutes after Cr2O3 inhalation, FEV1.0 had decreased by 32%. Chest X-ray film revealed no abnormalities. Histological findings of the lung tissue obtained by open lung biopsy revealed bronchospasm and peri-bronchial inflammatory cell infiltration without bronchiolar obstruction, and thus bronchiolitis obliterans was ruled out. This patient fulfilled the criteria of reactive airway dysfunction syndrome (RADS) proposed by Brooks; onset of symptoms very soon after a single exposure to an irritant, persistence of asthmatic symptoms, obstructive pulmonary dysfunction, and the presence of airway hyperreactivity to methacholine. During the initial hospital stay, asthmatic symptoms were relieved by intensive anti-asthmatic treatment including oral and parenteral corticosteroids. However, the patient's asthmatic symptoms have lasted for eight years and necessitate the use of systemic steroids, and regular use of inhaled steroids and bronchodilators. We know of no previous reported case of RADS after chromate exposure.

[Three cases of pneumothorax due to lung cancer].

Spontaneous pneumothorax due to primary lung cancers are increasingly reported as of lung cancer incidence increased. We experienced one lung cancer case with pneumothorax as a primary symptom and two who developed pneumothorax after chemotherapy, among 238 cases of primary lung cancer. In the literature, 76 cases of pneumothorax due to primary lung cancers are described. The mechanism of pneumothorax is discussed.