Oxidative stress potentially enhances FcεRI-mediated leukotriene C4 release dependent on the late-phase increase of intracellular glutathione in mast cells.

Cysteinyl leukotrienes (cysLTs), which include leukotriene C4 (LTC4), are the predominant class of LTs synthesized by mast cells. CysLTs can induce many of the abnormalities seen in asthma. LTC4 is generated by the conjugation of LTA4 with reduced glutathione (GSH) by LTC4 synthase. During screening of the effects of prostanoids on high-affinity IgE receptor (FcεRI)-mediated LTC4 release from mast cells, we realized that some prostanoids, including ONO-AE1-259-01 and ONO-AE-248, inhibited LTC4 release, which was associated with a decrease in the amount of intracellular total GSH. We ascertained that l-buthionine-S,R-sulfoximine (BSO), a selective inhibitor of glutamate-cysteine ligase, inhibited LTC4 release. In addition, cell-permeable GSH, the glutathione reduced form ethyl ester (GSH-OEt), enhanced LTC4 release in accordance with the change in intracellular total GSH. Depletion of intracellular total GSH induced by ONO-AE-248 or BSO enhanced FcεRI-mediated LTB4 release in contrast to LTC4. Oxidative stress contributes to many pathological conditions including asthma. GSH is a major soluble antioxidant and a cofactor for several detoxifying enzymes including GSH peroxidase. Exposure of mast cells to hydrogen peroxide (H2O2) or diamide to mimic oxidative stress unexpectedly increased rather than decreased the intracellular reduced GSH content as well as total GSH in the late phase (i.e., 24 or 48 h after exposure), which was accompanied by an increase in LTC4 release. In conclusion, FcεRI-mediated LTC4 release from mast cells is mainly regulated by the amount of intracellular GSH. In some cases, oxidative stress may induce a late-phase increase in intracellular GSH, resulting in enhanced LTC4 release from mast cells.

[Pulmonary nocardiosis with elevation of serum beta-D-glucan in a patient with polymyositis].

A 73-year-old woman with polymyositis, who had received corticosteroids and immune-suppressive agents, was admitted to our hospital because of general fatigue and severe cough. Chest X-ray film and CT scan showed a large tumor shadow in the left upper lobe and several ground-glass opacities (GGOs) scattered in both lungs. As the white blood cell and C-reactive protein levels were elevated, pnueumonia was suspected and antibiotics were administered. Subsequently, Nocardia spp. was cultured from the sputum and pulmonary nocardiosis was established. She gradually recovered after sulfamethoxazole-trimethoprim (ST) administration. The pretreatment serum beta-D-glucan level was highly elevated and decreased in parallel with clinical feature. In general, ST should be administered for 6 months to treat pulmonary nocardiosis in a compromised host. It is possible that P3-D-glucan may be a useful marker to treat pulmonary nocardiosis in patients with polymyositis.

JNK1 and JNK2 differently regulate IL-12 production in THP-1 macrophage cells.

Macrophages play a key role in initiating the innate responses to infection by secreting cytokines such as interleukin-12 (IL-12). This study defined the distinct regulation of lipopolysaccharide (LPS)-mediated IL-12 production by c-jun NH(2)-terminal kinase (JNK)1 and JNK2 isoforms in human macrophages. Knockdown of JNK1 and JNK2 by small interference RNA (siRNA) reduced and enhanced LPS-induced IL-12 p40 production in THP-1 macrophage cells, respectively. The simultaneous knockdown of JNK1 and JNK2 augmented LPS-induced IL-12 production as well as a specific JNK inhibitor. In addition, transfection of siRNA against phosphoinositide 3-kinase (PI3K) p110beta attenuated LPS-induced IL-12 production and JNK1 phosphorylation, while not affecting JNK2 phosphorylation. These findings indicate that JNK1- and JNK2-mediated signaling plays a positive and a negative role, respectively, in LPS-induced IL-12 production and PI3K p110beta controls LPS-induced JNK1 activation, not JNK2 activation, resulting in the positive regulation of IL-12 production in THP-1 macrophage cells.

[A case of cryptogenic organizing pneumonia with eosinophilic infiltration which was difficult to distinguish from nonspecific interstitial pneumonia].

A 57-year-old man presented with shortness of breath of four months duration which had recently become worse. A chest X-ray and computed tomography (CT) showed diffuse ground-glass opacities of the bilateral lower lungs suggesting interstitial pneumonia. The number of eosinophils was increased in the bronchoalveolar lavage fluid (BALF) (52%) and peripheral blood. A histological examination of the specimen obtained by TBLB revealed organized pneumonia with slight infiltration of inflammatory cell. Because the images were not typical of chronic eosinophilic pneumonia, video-assisted thoracic surgery biopsy was performed. The histological findings of the resected specimen showed organizing pneumonia with infiltration of eosinophils in the alveolar walls. He had not taken any medication prior to coming to the hospital and he was negative for medicine-related pneumonia. The oral administration of prednisolone (0.5 mg/kg) improved his symptoms and also CT findings.

PI3K p110beta positively regulates lipopolysaccharide-induced IL-12 production in human macrophages and dendritic cells and JNK1 plays a novel role.

The PI3K family is thought to participate in TLR signaling, and it has been reported to be a negative regulator of TLR-mediated production of IL-12, a key inducer of Th1 responses. However, the role of individual PI3K subtypes in IL-12 production remains obscure. We defined the distinct regulation of LPS-mediated IL-12 production by p110alpha and p110beta catalytic subunits of PI3K in human APCs. We observed that knockdown of PI3K p110beta by small interfering RNA (siRNA) suppressed both LPS-induced IL-12 protein production and mRNA expression in monocyte-derived macrophages and dendritic cells (DCs). Knockdown of PI3K p110alpha by siRNA reduced LPS-induced IL-12 protein production in both cell types. Conversely, knockdown of PI3K p110alpha suppressed LPS-induced IL-12 mRNA expression in monocyte-derived macrophages but minimally affected monocyte-derived DCs. PI3K p110beta siRNA inhibited JNK activation, but not p38 MAPK or ERK activation, stimulated by LPS, while PI3K p110alpha siRNA did not affect LPS-induced JNK, p38 MAPK, or ERK activation in both cell types. Transfection of siRNA against JNK1, JNK2, and both decreased LPS-induced IL-12 production. Furthermore, PI3K p110beta siRNA attenuated LPS-induced JNK1 phosphorylation, while not affecting JNK2 phosphorylation. Our findings indicate that PI3K p110beta positively controls LPS-induced IL-12 production through the JNK1-dependent pathway in human macrophages and DCs.

[Effective gemcitabine plus S-1 treatment in elderly non-small cell lung cancer patients with multiple bone metastases].

An 81-year-old man presented with right shoulder pain 10 months after surgery for non-small cell lung cancer (p- T3N0M0, Stage II b). FDG-PET revealed an increased uptake in the multiple bone and mediastinal lymph nodes, suggestive of recurrence of lung cancer. He was given combined chemotherapy of gemcitabine plus S-1 as first-line treatment. This regimen was performed as a phase I trial for an elderly patient with advanced non-small cell lung cancer, and the treatment proved effective. No toxic events were observed due to this therapy, so he was treated as an outpatient for one year. He was considered to have good quality of life.

Glutathione redox regulates TGF-beta-induced fibrogenic effects through Smad3 activation.

Transforming growth factor-beta (TGF-beta) plays a pivotal role in the fibrogenic action involved in the induction of connective tissue growth factor (CTGF), extracellular matrix and fibroblast transformation. Smad3 mediates TGF-beta signaling related to the fibrotic response. In human lung fibroblasts or bronchial smooth muscle cells, we demonstrated that an increase in the intracellular glutathione level suppressed TGF-beta1-induced phosphorylation of Smad3, while inhibiting TGF-beta1-induced expressions of CTGF, collagen type1, fibronectin and transformation into myofibroblasts, which are characterized by the expression of alpha-smooth muscle actin. These data indicate that the intracellular glutathione redox status regulates TGF-beta-induced fibrogenic effects through Smad3 activation.

Phase I study of oral S-1 plus Cisplatin with concurrent radiotherapy for locally advanced non-small-cell lung cancer.

PURPOSE: To determine the maximum tolerated dose (MTD) and recommended dose (RD) of S-1 in combination with cisplatin and thoracic radiotherapy in patients with unresectable Stage III non-small-cell lung cancer (NSCLC). METHODS AND MATERIALS: S-1 was administered orally twice daily for 14 days and cisplatin on Days 1 and 8 of each cycle; this was repeated every 3 weeks. Doses of each drug were planned as follows: level 0, 50/40; level 1, 60/40; level 2, 70/40; level 3, 80/40 (S-1 [mg/m(-2)/day(-1)]/cisplatin [mg/m(-2)/day(-1)]). Thoracic radiation therapy was administered in 2 Gy fractions five times weekly to a total dose of 60 Gy. RESULTS: Ten patients were enrolled in this study. All patients received 60 Gy of thoracic radiotherapy and 7 (70%) patients received four cycles of chemotherapy. At level 1, 2 of 3 patients experienced a delay exceeding 10 days in the cisplatin administration of Day 29. Grade 4 neutropenia and Grade 3 fever occurred in 1 and 1 patients, respectively. Nonhematologic toxicities were mild. None developed >or=Grade 3 esophagitis or lung toxicity. At level 0, 2 of 7 patients developed dose-limiting toxicity. Thus, level 1 was considered the MTD and Level 0 was selected as the RD. Objective responses were seen in all patients. CONCLUSIONS: The RD is the level 0 dose, and this regimen is a feasible and well-tolerated regimen for the treatment of patients with Stage III NSCLC.

[A case of elderly non-small cell lung cancer effectively treated by combined chemotherapy of gemcitabine and S-1].

The patient was an 86-year-old female who suffered from advanced non-small cell-lung cancer (cT4N3M1, Stage IV). She was treated with combined chemotherapy of gemcitabine and S-1 as first-line therapy. This regimen was performed as a phase I trial for elderly patients with advanced non-small cell lung cancer. The treatment assessment was evaluated as partial response. No toxic events were observed due to this therapy, so she was treated as an outpatient. She was considered to have good quality of life.

[A case of large cell neuroendocrine carcinoma in a patient with sarcoidosis].

A 60-year-old female smoker presented with bloody sputum and back pain. A computed tomographic (CT) scan of the chest revealed a mass lesion in S(1+2) of the left lung and hilarmediastinal lymphadenopathy. TBLB revealed small cell carcinoma. At first, we thought that the patient's clinical staging was c-T4N2M0 IIIB disease. However, it was pointed out she had had hilar-mediastinal lymphadenopathy 7 years previously. Though FDG-PET revealed an increased uptake in the hilar-mediastinal lymph node, she was determined to have surgery. The resected specimen revealed the swollen lymph nodes to be sarcoidosis, accompanying a large cell neuroendocrine carcinoma (LCNEL). She was treated with a combination chemotherapy consisting of cisplatin and VP-16 and radiotherapy concurrently. Chest CT revealed a partial response of the primary site after 3 courses of chemoradiotherapy. Information on concomitant malignancies accompanying sarcoidosis is limited. Although the main types of concomitant malignancy are lung cancer, and lymphoma, there has been no description of LCNEC as a concomitant malignancy. In sarcoidosis patients, clinicians should be alert to the possibility of concomitant malignancy.

Phase I trial of oral S-1 plus gemcitabine in elderly patients with nonsmall cell lung cancer.

We conducted a phase I study to determine the maximum tolerated dose, the recommended dose and the safety profile of S-1 and gemcitabine combination regimen in the treatment of elderly patients (> or = 70 years) with advanced nonsmall cell lung cancer (NSCLC). Chemotherapy-naive patients with advanced NSCLC were treated with S-1 and gemcitabine. S-1 was administered orally twice daily for 14 days and gemcitabine on days 1 and 15 of each cycle, and this was repeated every 4 weeks. Doses of each drug were planned as follows: level 1, 800/60; level 2, 1000/60; level 3, 1000/70; and level 4, 1000/80 [gemcitabine (mg/m2/ day)/S-1 (mg/m2/day)]. The dose-limiting toxicity (DLT) of the regimen was assessed during the first chemotherapy cycle. Sixteen patients were enrolled in this study. The main grade 3 toxicities observed during the first cycle were neutropenia (43.7%), leukopenia (18.7%), and hyperglycemia. One of six patients in level 3 had DLT. Although no patients in level 4 experienced DLT, this level was considered the maximum tolerated dose. Level 4 was selected as the recommended dose. Objective responses were seen in four patients (response rate, 42.9%). The combination of S-1 plus gemcitabine is a feasible and well-tolerated regimen for the treatment of elderly patients with advanced NSCLC.

Resolvin E1 dampens airway inflammation and hyperresponsiveness in a murine model of asthma.

Resolvin E1 (RvE1; 5S, 12R, 18R-trihydroxyeicosapentaenoic acid) is an anti-inflammatory lipid mediator derived from the omega-3 fatty acid eicosapentaenoic acid (EPA). It has been recently shown that RvE1 is involved in the resolution of inflammation. However, it is not known whether RvE1 is involved in the resolution of asthmatic inflammation. To investigate the anti-inflammatory effect of RvE1 in asthma, a murine model of asthma was studied. After RvE1 was administered to mice intraperitoneally, there were decreases in: airway eosinophil and lymphocyte recruitment, specific Th2 cytokine, IL-13, ovalbumin-specific IgE, and airway hyperresponsiveness (AHR) to inhaled methacholine. Moreover, RvE1-treated mice had significantly lower mucus scores compared to vehicle-treated mice based on the number of goblet cells stained with periodic acid-schiff (PAS). These findings provide evidence that RvE1 is a pivotal counterregulatory signal in allergic inflammation and offer novel multi-pronged therapeutic approaches for human asthma.

Proton pump inhibitor improves breath marker in moderate asthma with gastroesophageal reflux disease.

BACKGROUND: Gastroesophageal reflux disease (GERD) influences the symptoms of asthma with acid and oxidative stress. OBJECTIVES: The purpose of this study was to determine the usefulness of measurement of the acid stress marker pH and the oxidative stress marker 8-isoprostane by exhaled breath condensate in proton pump inhibitor (PPI) therapy effect on moderate asthma patients with GERD. METHODS: The pH and the concentration of 8-isoprostane were measured in the exhaled breath condensate of patients with moderate asthma (n = 36) and healthy subjects (n = 26). Two months of PPI therapy (lansoprazole at 30 mg/day) were done in the asthma patients with (n = 13) or without (n = 13) GERD according to a questionnaire for the diagnosis of reflux disease, and exhaled markers were measured. RESULTS: The pH was lower (7.3 +/- 0.3) and the 8-isoprostane level was higher (27.7 +/- 2.3) in the asthma patients than in the healthy control subjects (pH 7.5 +/- 0.2 and 8-isoprostane 6.6 +/- 1.2). Two months of PPI therapy improved the pH (from 7.2 +/- 0.1 to 7.3 +/- 0.1) and the 8-isoprostane concentration (from 32.7 +/- 3.4 to 19.2 +/- 3.4) in the asthma patients with GERD, along with improvement of GERD symptoms. However, these markers did not change in the asthma patients without GERD. CONCLUSIONS: Measurement of the pH and 8-isoprostane level of exhaled breath condensate may be useful to evaluate the influence of GERD on asthma, as well as to determine the timing of intermittent PPI therapy.

Rac1 negatively regulates lipopolysaccharide-induced IL-23 p19 expression in human macrophages and dendritic cells and NF-kappaB p65 trans activation plays a novel role.

IL-23 is a heterodimeric cytokine composed of a unique p19 subunit and of a p40 subunit that is also common to IL-12. We defined the distinct signaling mechanisms that regulate the LPS-mediated induction of IL-23 p19 and p40 in human macrophages and dendritic cells. We found that the overexpression of dominant-negative Rac1 (N17Rac1) enhanced LPS-induced IL-23 p19 expression but did not alter p40 expression or IL-12 p70 production in PMA-treated THP-1 macrophages and in human monocyte-derived dendritic cells. Although the inhibition of either p38 MAPK or JNK enhanced LPS-induced p19 expression, N17Rac1 did not influence either p38 MAPK or JNK activation. By contrast, N17Rac1 augmented both NF-kappaB gene expression and p65 trans activation stimulated by LPS without affecting the degradation of IkappaB-alpha or DNA binding to NF-kappaB. Furthermore, small interference RNA of NF-kappaB p65 attenuated cellular amounts of p65 and suppressed LPS-induced p19 expression but did not affect p40 expression. Our findings indicate that Rac1 negatively controls LPS-induced IL-23 p19 expression through an NF-kappaB p65 trans activation-dependent, IkappaB-independent pathway and that NF-kappaB p65 regulates LPS-induced IL-23 p19, but not p40, expression, which causes differences in the control of IL-23 p19 and p40 expression by Rac1.

Sphingosine 1-phosphate inhibits migration and RANTES production in human bronchial smooth muscle cells.

Sphingosine 1-phosphate (S1P), a bioactive lipid mediator, has been shown to be increased in bronchoalveolar lavage fluid after allergen challenge in asthmatic patients. Here, we examined S1P actions and their intracellular signalings in cultured human bronchial smooth muscle cells (BSMCs). Expression of mRNAs of three subtypes of S1P receptors, including S1P(1), S1P(2), and S1P(3), was detected in BSMCs, and exposure of the cells to S1P inhibited platelet-derived growth factor (PDGF)-induced migration and tumor necrosis factor-alpha-induced RANTES production. S1P also inhibited PDGF-induced Rac1 activation, and dominant negative Rac1 inhibited PDGF-induced migration. On the other hand, dominant negative Galpha(q) attenuated the S1P-induced inhibition of RANTES production. Finally, an S1P(2)-selective antagonist, JTE-013, suppressed the S1P-induced inhibition of migration response and RANTES production. These results suggest that S1P attenuates cell migration by inhibiting a Rac1-dependent signaling pathway and decreases RANTES production by stimulating a Galpha(q)-dependent mechanism both possibly through the S1P(2) receptors.

Safety and efficacy of extracorporeal granulocyte and monocyte adsorption apheresis in patients with severe persistent bronchial asthma.

The Adacolumn is an adsorptive-type extracorporeal device, which is filled with cellulose diacetate beads that selectively adsorb granulocytes and monocytes. Patients with severe persistent asthma experience highly variable continuous symptoms and severe exacerbations in spite of medication based on inhaled glucocorticosteroids. Granulocyte and monocyte adsorption apheresis using extracorporeal circulation through the Adacolumn was performed in nine patients with severe persistent asthma. The extracorporeal circulation through the Adacolumn was performed once a week for 5 weeks. We were able to perform this therapy without any severe adverse effects in all patients, although one patient complained of general fatigue just after the circulations. In six of the nine patients, the increase in peak expiratory flow (PEF) was more than 50 mL/min. The average increase in morning PEF was 23.3% while that in the evening PEF was 26.4% after the therapy. This therapy was not harmful for patients with severe persistent asthma. A placebo-controlled study will be desired to evaluate the efficacy of this nonpharmacological strategy accurately.

KL-6-producing invasive thymoma.

We report a patient with KL-6-producing invasive thymoma. A 58-year-old man was admitted complaining of dyspnea and fatigability. Computed tomography of the chest revealed interstitial pneumonia and an anterior mediastinal tumor. The tumor was surgically extirpated and diagnosed as invasive thymoma. Serum KL-6 levels later increased further and another tumor was found in the liver. That liver tumor was resected and histologically diagnosed as a metastasis of thymoma. Following resection, the serum KL-6 level decreased. Tumor cells of both primary and metastatic lesions exhibited positive reactivity to immunohistochemical staining for KL-6. A review of this case is presented.