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IL-17A and IL-17F are both involved in the pathogenesis of neutrophilic inflammation observed in COPD and severe asthma. To explore this, mice deficient in both Il17a and Il17f and wild type (WT) mice were exposed to cigarette smoke or environmental air for 5 to 28 days and changes in inflammatory cells in bronchoalveolar lavage (BAL) fluid were determined. We also measured the mRNA expression of keratinocyte derived chemokine (Kc), macrophage inflammatory protein-2 (Mip2), granulocyte–macrophage colony stimulating factor (Gmcsf) and matrix metalloproteinase-9 (Mmp9 ) in lung tissue after 8 days, and lung morphometric changes after 24 weeks of exposure to cigarette smoke compared to air-exposed control animals. Macrophage counts in BAL fluid initially peaked at day 8 and again on day 28, while neutrophil counts peaked between day 8 and 12 in WT mice. Mice dual deficient with Il17a and 1l17f showed similar kinetics with macrophages and neutrophils, but cell numbers at day 8 and mRNA expression of Kc, Gmcsf and Mmp9 were significantly reduced. Furthermore, airspaces in WT mice became larger after cigarette smoke exposure for 24 weeks, whereas this was not seen dual Il17a and 1l17f deficient mice. Combined Il17a and Il17f deficiency resulted in significant attenuation of neutrophilic inflammatory response and protection against structural lung changes after long term cigarette smoke exposure compared with WT mice. Dual IL-17A/F signalling plays an important role in pro-inflammatory responses associated with histological changes induced by cigarette smoke exposure.

The present study investigated the cancer prevalence and anticancer treatment patients undergoing hemodialysis at Nagoya Memorial Hospital. We retrospectively analyzed 663 patients undergoing hemodialysis between September 2014 and August 2019, including patient characteristics such as age, sex, and underlying diseases, cancer type, and cancer treatment. Seventy-eight patients (11.9%) of the dialysis population were diagnosed with cancer. Cancer type was then compared between registered cancer patients undergoing maintenance dialysis (N = 78) and non-dialysis controls (N = 3279) during the same period. Colorectal carcinoma is the most common malignancy diagnosed in our hospital, accounting for approximately 15% of all types of cancers. The data of anticancer treatment for this disease were compared between dialysis patients (N = 15) and controls (N = 563), whose clinical stages were defined according to the Japanese Classification of Colorectal, Appendiceal, and Anal Carcinoma. Since the need to administer chemotherapeutic agents to dialysis patients with colorectal carcinoma will increase, oncologists should collaborate with nephrologists to cautiously manage anticancer treatment to avoid severe toxicities.

OBJECTIVES This study aimed to evaluate the safety and reliability of percutaneous computed tomography (CT)-guided lipiodol marking for undetectable pulmonary lesions before video-assisted thoracic surgery (VATS). METHODS We retrospectively analysed the cases of CT-guided lipiodol marking followed by VATS in 9 institutes from May 2006 to March 2018. Lipiodol (0.2–0.5 ml) was percutaneously injected closely adjacent to undetectable pulmonary lesions with computed-tomography guidance. Lipiodol spots were identified using C-arm-shaped fluoroscopy during VATS. We grasped the lipiodol spots, including the target lesions, with ring-shaped forceps and resected them. RESULTS Of 1182 lesions, 1181 (99.9%) were successfully marked. In 1 case, the injected lipiodol diffused, and no spot was created. Of the 1181 lesions, 1179 (99.8%) were successfully resected with intraoperative fluoroscopy. Two lipiodol spots were not detected because of the lipiodol distribution during the division of pleural adhesions. The mean lesion size was 9.1 mm (range 1–48 mm). The mean distance from the pleural surface was 10.2 mm (range 0–43 mm). Lipiodol marking-induced pneumothorax occurred in 495 (57.1%) of 867 cases. Of these, chest drainage was required in 59 patients (6.8%). The other complications were 19 (2.2%) cases of bloody sputum, 3 (0.35%) cases of intravascular air, 1 (0.12%) case of pneumonia and 1 (0.12%) case of cerebral infarction. There were no lipiodol marking-induced deaths or sequelae. CONCLUSIONS Preoperative CT-guided lipiodol marking followed by VATS resection was shown to be a safe and reliable procedure with a high success rate and acceptably low severe complication rate.

In patients with asthma, the inhalation of elevated amounts of fungal spores and hyphae may precipitate the onset of asthma or worsen control to the extent of being life-threatening. Sensitisation to fungi, especially Aspergillus fumigatus, is found in 15% to 48% of asthmatics in secondary care and is linked to worse asthma control, hospitalisation, bronchiectasis and fixed airflow obstruction, irrespective of whether allergic bronchopulmonary aspergillosis (ABPA) is diagnosed. ABPA represents a florid response to the presence of Aspergillus spp. but up to 70% of patients with severe asthma exhibit sensitisation to different fungi without meeting the diagnostic criteria for ABPA. The presence of persistent endobronchial colonisation with fungi, especially A. fumigatus, is linked to significantly higher rates of radiological abnormalities, lower post-bronchodilator FEV1 and significantly less reversibility to short acting bronchodilators. The therapeutic benefit for antifungal intervention in severe asthma is based on the assumption that reductions in airway fungal burden may result in improvements in asthma control, lung function and symptoms (especially cough). This contention is supported by several prospective studies which demonstrate the effectiveness of antifungals for the treatment of ABPA. Significantly, these studies confirm lower toxicity of treatment with azoles versus high dose oral corticosteroid dosing regimens for ABPA. Here we review recent evidence for the role of fungi in the progression of severe asthma and provide recommendations for the use of antifungal agents in patients with severe asthma, airways fungal infection (mycosis) and fungal colonisation. Documenting fungal airways colonisation and sensitisation in those with severe asthma opens up alternative therapy options of antifungal therapy, which may be particularly valuable in low resource settings.

Although Candida spp. are frequently detected in fungal cultures of respiratory secretions, their presence is normally assumed to reflect benign colonization. However, there is growing evidence that Candida spp. is involved in pathogenesis of respiratory diseases such as bronchiectasis, asthma and COPD. Here the aim is to establish a Candida albicans lung infection model in mice, and investigate the effects of PC945, a novel antifungal triazole optimised for lung delivery. Candida albicans (MYA4901) was administered intranasally to immunocompromised temporarily neutropenic A/J mice, pre-treated with hydrocortisone and cyclophosphamide. In this model, 71 % of mice were dead 5 days post inoculation. High levels of fungal burden in lung (6.55 ± 0.20 Log CFU/g of lung) and inflammatory markers CXCL1 in BALF (5.3 ± 1.1 ng/mL) were observed on day 5. Histology revealed significant inflammation and acute lung injury. PC945 saline suspension, when treated intranasally once daily, starting a day after candida inoculation, improved survival rates and inhibited lung fungal burden as well as BALF CXCL1 in a dose-dependent manner (0.56, 2.8 and 14 µg/mouse). Thus, Candida albicans, when inoculated intranasally, induced acute lung injury/death, and intranasally dosed PC945 showed potent antifungal effects. Therefore, PC945 has the potential to be a novel inhaled therapy for the treatment of C. albicans infection/colonization in humans.

Nontypeable Haemophilus influenzae (NTHi) is reported to be colonised more in the lower airways of COPD and asthma patients. The colonization is known to be associated with considerable airway inflammation and cause increased frequency of exacerbations as well as lung cancer. Treatment of respiratory tract infection with NTHi is often only partially successful with ongoing infection and inflammation. Successful treatment will be dependent on a better understanding of the pathogenesis and pharmacological response to this bacterium although there is no or little NTHi airway infection model reported. Therefore, aim of this study is to establish NTHi airway infection model in mice. NTHi (ATCC49766) at sub-lethal concentration was administered intranasally to intact or immunocompromised (IC) A/J mice, pre-treated with hydrocortisone and cyclophosphamide. BALF and lung were collected a day after bacteria intranasal inoculation, and the bacterial burden and biomarkers were evaluated. The IC mice showed more than 50-fold higher bacteria burden in BALF (geometric mean 3.6 Log, CFU/mL in intact mice vs 5.4 Log, CFU/mL in IC mice) and lung (4.5 Log, CFU/g in intact mice vs 6.4 Log, CFU/g in IC mice), and showed much higher levels of biomarkers CXCL1 and malonaldehyde in BALF. Levofloxacin, when dosed intranasally or orally, inhibited bacterial burden completely. Thus, we were able to establish NTHi airway infection model, which can be used to understand pathogenesis/host response to NTHi and to identify new therapy for NTHi airway infection.

The prevalence of asthma has increased worldwide. Asthma exacerbations triggered by upper respiratory tract viral infections remain a major clinical problem and account for hospital admissions and time lost from work. Virus-induced asthma exacerbations cause airway inflammation, resulting in worsening asthma and deterioration in the patients’ quality of life, which may require systemic corticosteroid therapy. Despite recent advances in understanding the cellular and molecular mechanisms underlying asthma exacerbations, current therapeutic modalities are inadequate for complete prevention and treatment of these episodes. The pathological role of cellular senescence, especially that involving the silent information regulator 2 homolog sirtuin (SIRT) protein family, has recently been demonstrated in stable and exacerbated chronic respiratory disease states. This review discusses the role of SIRT1 in the pathogenesis of bronchial asthma. It also discusses the role of SIRT1 in inflammatory cells that play an important role in virus-induced asthma exacerbations. Recent studies have hypothesized that SIRT1 is one of major contributors to cellular senescence. SIRT1 levels decrease in Th2 and non-Th2-related airway inflammation, indicating the role of SIRT1 in several endotypes and phenotypes of asthma. Moreover, several models have demonstrated relationships between viral infection and SIRT1. Therefore, targeting SIRT1 is a novel strategy that may be effective for treating virus-induced asthma exacerbations in the future.

Polymorphism at the 17q21 gene locus and wheezing responses to rhinovirus (RV) early in childhood conspire to increase the risk of developing asthma. However, the mechanisms mediating this gene–environment interaction remain unclear. In this study, we investigated the impact of one of the 17q21-encoded genes, ORMDL3 (orosomucoid-like 3), on RV replication in human epithelial cells. ORMDL3 knockdown inhibited RV-A16 replication in HeLa, BEAS-2B, A549, and NCI-H358 epithelial cell lines and primary nasal and bronchial epithelial cells. Inhibition varied by RV species, as both minor and major group RV-A subtypes RV-B52 and RV-C2 were inhibited but not RV-C15 or RV-C41. ORMDL3 siRNA did not affect expression of the major group RV-A receptor ICAM-1 or initial internalization of RV-A16. The two major outcomes of ORMDL3 activity, SPT (serine palmitoyl-CoA transferase) inhibition and endoplasmic reticulum (ER) stress induction, were further examined: silencing ORMDL3 decreased RV-induced ER stress and IFN-β mRNA expression. However, pharmacologic induction of ER stress and concomitant increased IFN-β inhibited RV-A16 replication. Conversely, blockade of ER stress with tauroursodeoxycholic acid augmented replication, pointing to an alternative mechanism for the effect of ORMDL3 knockdown on RV replication. In comparison, the SPT inhibitor myriocin increased RV-A16 but not RV-C15 replication and negated the inhibitory effect of ORMDL3 knockdown. Furthermore, lipidomics analysis revealed opposing regulation of specific sphingolipid species (downstream of SPT) by myriocin and ORMDL3 siRNA, correlating with the effect of these treatments on RV replication. Together, these data revealed a requirement for ORMDL3 in supporting RV replication in epithelial cells via SPT inhibition.

Background: Sestrin 2, Endocan, and Sirtuin 1 are distinct molecules with some biologic actions associated with asthma pathophysiology. The aim of the present study was to determine the molecular level differences attributable to underlying asthma severity. Methods: We initially recruited 85 asthmatics with a wide spectrum of severity. All of the patients were optimally treated according to current guidelines. Demographics, test results of lung function, and treatment regimes of all patients were recorded. Sestrin 2, Endocan, and Sirtuin 1 were measured in different biological samples (sputum with two processing methods and serum). Results: A total of 60 patients (35 with severe asthma) were analyzed, since 25 patients failed to produce an adequate sample of sputum. Patients with severe asthma showed significantly higher values for Sestrin 2 [pg/mL], measured in both sputum supernatant and cell pellet, compared to those with mild to moderate asthma [9524 (5696, 12,373) vs. 7476 (4265, 9273) p = 0.029, and 23,748 (15,280, 32,742) vs. 10,084 (3349, 21,784), p = 0.008, respectively]. No other significant differences were observed. No significant associations were observed between biomarkers, inflammatory cells, and lung function. Conclusion: Sestrin 2 is increased in patients with severe asthma as part of a mechanism that may modify structural alterations through the imbalance between oxidative stress and antioxidant activity.

The purpose of this study was to clarify the clinical efficacy of postoperative non-invasive positive pressure ventilation (PONIV) after pulmonary lobectomy in patients with lung cancer. From August 2010 and July 2015, 143 patients with lung cancer who underwent pulmonary lobectomy were retrospectively reviewed. PONIV was used immediately after surgery until the morning of postoperative day (POD) 1. Arterial blood gas was analyzed before and just after surgery (POD0) and on POD1. Oxygenation ability was perioperatively assessed by PaO2/FiO2 ratio, alveolar–arterial oxygen difference (A-aDO2), and respiratory index (A-aDO2/PaO2). 112 patients received PONIV. From POD0 to POD1, the PaO2/FiO2 ratio significantly improved in all patients who received PONIV (333 ± 83 to 359 ± 47 mmHg, p = 0.004). Moreover, A-aDO2 and respiratory index significantly decreased following PONIV. PONIV significantly improved the PaO2/FiO2 ratio in patients with PaO2/FiO2 ratio of ≤ 300 on POD0, older age (≥ 70 years), higher body mass index (≥ 25 kg/m2), and longer one-lung ventilation time (≥ 180 min). There was no respiratory failure requiring mechanical ventilation and no mortality. PONIV effectively improved oxygenation in patients undergoing pulmonary lobectomy in patients with poor status, especially in patients with PaO2/FiO2 ratio of ≤ 300 on POD0. PONIV could be an option of perioperative management for major thoracic surgery.

Summary We used nasosorption as a noninvasive method of respiratory mucosal sampling in a paediatric virology setting. Nasosorption was well tolerated and demonstrated respiratory syncytial virus load to be associated with disease severity and elevated mediators of inflammation., Background. Existing respiratory mucosal sampling methods are flawed, particularly in a pediatric bronchiolitis setting. Methods. Twenty-four infants with bronchiolitis were recruited: 12 were respiratory syncytial virus (RSV)–positive, 12 were RSV-negative. Infants were sampled by nasosorption and nasopharyngeal aspiration (NPA). Results. Nasosorption was well tolerated and identified all RSV+ samples. RSV load measured by nasosorption (but not NPA) correlated with length of hospital stay (P = .04) and requirement for mechanical ventilation (P = .03). Nasosorption (but not NPA) levels of interferon γ, interleukin 1β, CCL5/RANTES, and interleukin 10 (IL-10) were elevated in RSV+ bronchiolitis (all P < .05), furthermore CCL5 and IL-10 correlated with RSV load (P < .05). Conclusions. Nasosorption allowed measurement of RSV load and the mucosal inflammatory response in infants.

Background and Purpose To investigate whether a narrow spectrum kinase inhibitor RV1088, which simultaneously targets specific MAPKs, Src and spleen tyrosine kinase (Syk), is more effective at inhibiting inflammatory signalling in rheumatoid arthritis (RA) than single kinase inhibitors (SKIs). Experimental Approach elisas were used to determine the efficacy of RV1088, clinically relevant SKIs and the pharmaceutical Humira on pro-inflammatory cytokine production by activated RA synovial fibroblasts, primary human monocytes and macrophages, as well as spontaneous cytokine synthesis by synovial membrane cells from RA patients. In human macrophages, RNAi knockdown of individual kinases was used to reveal the effect of inhibition of kinase expression on cytokine synthesis. Key Results RV1088 reduced TNF-α, IL-6 and IL-8 production in all individual activated cell types with low, nM, IC50s. SKIs, and combinations of SKIs, were significantly less effective than RV1088. RNAi of specific kinases in macrophages also caused only modest inhibition of pro-inflammatory cytokine production. RV1088 was also significantly more effective at inhibiting IL-6 and IL-8 production by monocytes and RA synovial fibroblasts compared with Humira. Finally, RV1088 was the only inhibitor that was effective in reducing TNF-α, IL-6 and IL-8 synthesis in RA synovial membrane cells with low nM IC50s. Conclusions and Implications This study demonstrates potent anti-inflammatory effect of RV1088, highlighting that distinct signalling pathways drive TNF-α, IL-6 and IL-8 production in the different cell types found in RA joints. As such, targeting numerous signalling pathways simultaneously using RV1088 could offer a more powerful method of reducing inflammation in RA than targeting individual kinases.

Corticosteroid insensitive airway inflammation is one of major barrier to effective managements of chronic airway diseases, such as chronic obstructive pulmonary disease (COPD) and severe asthma. The role of nonreceptor tyrosine kinase Src is important in airway inflammation in mice models of atopic asthma and COPD. Thus, in this study, we determined the effects of Src inhibitor, dasatinib, on airway inflammation induced by repeated intranasal exposure to lipopolysaccharide (LPS). Male mice (A/J strain, 5 weeks old) were intranasally exposed to LPS twice daily for 3 d, and dasatinib was intranasally treated 2 h prior to each LPS exposure. A day after the last stimulation, lungs and bronchoalveolar lavage fluid (BALF) were collected. Dasatinib attenuated the accumulation of inflammatory cells in lungs, and the increase in the numbers of inflammatory cells and the accumulation of cytokines/chemokines in BALF in a dose dependent manner. Therefore, this study suggested that targeting the Src can provide a new therapeutic approach for corticosteroid insensitive pulmonary diseases.

Background The detection of amyloid β accumulation might be useful in confirming the diagnosis of Alzheimer's disease in clinically suspected cases. However, confirmation of the disease by a positron emission tomography scan is not always available. Aim We measured the cerebral blood flow in clinically suspected Alzheimer's disease cases, and compared the findings between amyloid-positive and -negative cases. Methods At the Osaka City University Hospital, patients with “probable or possible Alzheimer's disease” were enrolled. In addition, “healthy volunteers” with no subjective decline in cognitive function or in cognitive tests were evaluated as controls. A positron emission tomography scan with Pittsburgh compound B as a tracer was utilized to detect amyloid accumulation. Results In the healthy control cases aged >60 years, a positive rate of amyloid increased steeply with age. In contrast, an amyloid-positive rate decreased with age in the clinically suspected Alzheimer's disease cases aged >60 years. The Mini-Mental State Examination score was higher and the disease duration was longer in the amyloid-negative group than in the positive group, suggesting a slower progression of dementia in the amyloid-negative patients, though the tendency failed to reach the statistical significance. Cerebral blood flow decrease was classified into four patterns. Typical Alzheimer's disease patterns were found exclusively in the amyloid-positive group, whereas frontal dominant and temporal dominant were more frequently found in the amyloid-negative group. Conclusion Even in the era of amyloid imaging, cerebral blood flow measurement can help differentiate Alzheimer's disease from other diseases causing apparent Alzheimer's disease-type cognitive impairment.

Objective: We aimed to analyze the clinical and histopathologic features of cancer-associated myositis (CAM) in relation to anti–transcriptional intermediary factor 1 γ antibody (anti-TIF1-γ-Ab), a marker of cancer association. Methods: We retrospectively studied 349 patients with idiopathic inflammatory myopathies (IIMs), including 284 patients with pretreatment biopsy samples available. For the classification of IIMs, the European Neuromuscular Center criteria were applied. Patients with CAM with (anti-TIF1-γ-Ab[+] CAM) and without anti-TIF1-γ-Ab (anti-TIF1-γ-Ab[−] CAM) were compared with patients with IIM without cancers within and beyond 3 years of myositis diagnosis. Results: Cancer was detected in 75 patients, of whom 36 (48%) were positive for anti-TIF1-γ-Ab. In anti-TIF1-γ-Ab(+) patients with CAM, cancers were detected within 1 year of myositis diagnosis in 35 (97%) and before 1 year of myositis diagnosis in 1. All the anti-TIF1-γ-Ab(+) patients with CAM satisfied the dermatomyositis (DM) criteria, including 2 possible DM sine dermatitis cases, and were characterized histologically by the presence of perifascicular atrophy, vacuolated fibers (VFs), and dense C5b-9 deposits on capillaries (dC5b-9). In contrast, 39 anti-TIF1-γ-Ab(−) patients with CAM were classified into various subgroups, and characterized by a higher frequency of necrotizing autoimmune myopathy (NAM). Notably, all 7 patients with CAM classified into the NAM subgroup were anti-TIF1-γ-Ab(−) and exhibited no dC5b-9 or VFs. Conclusions: CAM includes clinicohistopathologically heterogeneous disease entities. Among CAM entities, anti-TIF1-γ-Ab(+) CAM has characteristically shown a close temporal association with cancer detection and the histopathologic findings of dC5b-9 and VFs, and CAM with NAM is a subset of anti-TIF1-γ-Ab(−) CAM.

The discovery of a novel series of therapeutic agents that has been designed and optimized for treating chronic obstructive pulmonary disease is reported. The pharmacological strategy was based on the identification of compounds that inhibit a defined subset of kinase enzymes modulating inflammatory processes that would be effective against steroid refractory disease and exhibit a sustained duration of action after inhaled delivery.

Background Respiratory virus infections are commonly associated with COPD exacerbations, but little is known about the mechanisms linking virus infection to exacerbations. Pathogenic mechanisms in stable COPD include oxidative and nitrosative stress and reduced activity of histone deacetylase-2 (HDAC2), but their roles in COPD exacerbations is unknown. We investigated oxidative and nitrosative stress (O&NS) and HDAC2 in COPD exacerbations using experimental rhinovirus infection. Methods Nine subjects with COPD (Global Initiative for Chronic Obstructive Lung Disease stage II), 10 smokers, and 11 nonsmokers were successfully infected with rhinovirus. Markers of O&NS-associated cellular damage, and inflammatory mediators and proteases were measured in sputum, and HDAC2 activity was measured in sputum and bronchoalveolar macrophages. In an in vitro model, monocyte-derived THP-1 cells were infected with rhinovirus and nitrosylation and activity of HDAC2 was measured. Results Rhinovirus infection induced significant increases in airways inflammation and markers of O&NS in subjects with COPD. O&NS markers correlated with virus load and inflammatory markers. Macrophage HDAC2 activity was reduced during exacerbation and correlated inversely with virus load, inflammatory markers, and nitrosative stress. Sputum macrophage HDAC2 activity pre-infection was inversely associated with sputum virus load and inflammatory markers during exacerbation. Rhinovirus infection of monocytes induced nitrosylation of HDAC2 and reduced HDAC2 activity; inhibition of O&NS inhibited rhinovirus-induced inflammatory cytokines. Conclusions O&NS, airways inflammation, and impaired HDAC2 may be important mechanisms of virus-induced COPD exacerbations. Therapies targeting these mechanisms offer potential new treatments for COPD exacerbations.

Rationale: Respiratory syncytial virus (RSV) bronchiolitis is a major cause of morbidity and mortality in infancy. Severe disease is believed to result from uncontrolled viral replication, an excessive immune response, or both. Objectives: To determine RSV load and immune mediator levels in nasal mucosal lining fluid by serial sampling of nasal fluids from cases of moderate and severe bronchiolitis over the course of infection. Methods: Infants with viral bronchiolitis necessitating admission (n = 55) were recruited from a pediatric center during 2016 and 2017. Of these, 30 were RSV infected (18 “moderate” and 12 mechanically ventilated “severe”). Nasal fluids were sampled frequently over time using nasosorption devices and nasopharyngeal aspiration. Hierarchical clustering of time-weighted averages was performed to investigate cytokine and chemokine levels, and gene expression profiling was conducted. Measurements and Main Results: Unexpectedly, cases with severe RSV bronchiolitis had lower nasal viral loads and reduced IFN-γ and C-C chemokine ligand 5/RANTES (regulated upon activation, normal T cell expressed and secreted) levels than those with moderate disease, especially when allowance was made for disease duration (all P

Background Statins have pleiotropic immunomodulatory effects that may be beneficial in the treatment of asthma. We previously reported that treatment with atorvastatin improved asthma symptoms in smokers with asthma in the absence of a change in the concentration of a selection of sputum inflammatory mediators. Objective To determine the effects of atorvastatin alone and in combination with inhaled corticosteroid on a range of sputum cytokines, chemokines and growth factors implicated in the pathogenesis of asthma, and their association with asthma control questionnaire (ACQ) and/or asthma quality of life questionnaire (AQLQ) scores. Methods Sputum samples were analysed from a sub-group of 39 smokers with mild to moderate asthma recruited to a randomised controlled trial comparing atorvastatin (40 mg/day) versus placebo for four weeks, followed by inhaled beclometasone (400 μg/day) for a further four weeks. Induced sputum supernatant fluid was analysed (Luminex or biochemical analyses) for concentrations of 35 mediators. Results Sputum mediator concentrations were not reduced by inhaled beclometasone alone. Atorvastatin significantly reduced sputum concentrations of CCL7, IL-12p70, sCD40L, FGF-2, CCL4, TGF-α and MMP-8 compared with placebo and, when combined with inhaled beclometasone, reduced sputum concentrations of MMP-8, IL-1β, IL-10, MMP-9, sCD40L, FGF-2, IL-7, G-CSF and CCL7 compared to ICS alone. Improvements in ACQ and/or AQLQ scores with atorvastatin and ICS were associated with decreases in G-CSF, IL-7, CCL2 and CXCL8. Conclusion Short-term treatment with atorvastatin alone or in combination with inhaled beclometasone reduces several sputum cytokines, chemokines and growth factors concentrations unresponsive to inhaled corticosteroids alone, in smokers with asthma.

The rise in life expectancy worldwide has been accompanied by an increased incidence of age-related diseases, representing an enormous burden on healthcare services and society. All vital organs lose function with age, and this is well described in the lung, with a progressive decline in pulmonary function after the age of about 25 years. The lung ages, like any other organ, with progressive functional impairment and reduced capacity to respond to environmental stresses and injury. Normal physiological ageing results in enlarged alveolar spaces and loss of lung elasticity in the elderly known as 'senile emphysema', whereas in COPD there is destruction of the alveolar walls and fibrosis of peripheral airways. However, COPD shows striking age-associated features, such as an increase in cellular senescence, stem cell exhaustion, increased oxidative stress, alteration in the extracellular matrix and a reduction in endogenous antiageing molecules and protective pathways such as autophagy. In this review we discuss the evidence showing how oxidative stress induces accelerated ageing by upregulating the phosphatidylinositol-4,5-bisphosphate 3-kinase/AKT/mechanistic target of rapamycin signalling pathway resulting in depletion of stem cells, defective autophagy, reduced antioxidant responses and defective mitochondrial function thus generating further oxidative stress. Understanding the mechanisms of accelerated ageing in COPD may identify novel therapeutic approaches.

Although respiratory syncytial virus (RSV) is the most common cause of lower respiratory tract infection in infants and young children, attempts to develop an effective therapy have so far proved unsuccessful. Here we report the preclinical profiles of PC786, a potent nonnucleoside RSV L protein polymerase inhibitor, designed for inhalation treatment of RSV infection. PC786 demonstrated a potent and selective antiviral activity against laboratory-adapted or clinical isolates of RSV-A (50% inhibitory concentration [IC 50 ], 50 , 1.3 to 50.6 nM), which were determined by inhibition of cytopathic effects in HEp-2 cells without causing detectable cytotoxicity. The underlying inhibition of virus replication was confirmed by PCR analysis. The effects of PC786 were largely unaffected by the multiplicity of infection (MOI) and were retained in the face of established RSV replication in a time-of-addition study. Persistent anti-RSV effects of PC786 were also demonstrated in human bronchial epithelial cells. In vivo intranasal once daily dosing with PC786 was able to reduce the virus load to undetectable levels in lung homogenates from RSV-infected mice and cotton rats. Treatment with escalating concentrations identified a dominant mutation in the L protein (Y1631H) in vitro . In addition, PC786 potently inhibited RSV RNA-dependent RNA polymerase (RdRp) activity in a cell-free enzyme assay and minigenome assay in HEp-2 cells (IC 50 , 2.1 and 0.5 nM, respectively). Thus, PC786 was shown to be a potent anti-RSV agent via inhibition of RdRp activity, making topical treatment with this compound a novel potential therapy for the treatment of human RSV infections.

Background RSV is the most common cause of bronchiolitis in infants and is responsible for severe respiratory infections in the elderly and immunocompromised populations. RSV replicates in the columnar epithelial cells of the proximal and distal airways which are accessible to inhaled therapies. PC786 is a potent non-nucleoside RSV L-protein polymerase inhibitor designed for inhaled delivery. In preclinical studies, PC786 exhibits prolonged lung tissue residence with minimal systemic exposure, thus limiting the potential for adverse systemic effects. Methods A phase 1 study was conducted to evaluate the safety and pharmacokinetics of PC786 delivered in a suspension formulation by nebulizer (PARI LC SPRINT® device). Healthy volunteers (HVs) received placebo or PC786 as single ascending doses (0.5–20 mg, Cohort (C) 1), 5 mg BD for 7 days (C2), or 10 mg BD for 7 days (C3). Mild asthmatics received a single dose of PC786 5 mg or placebo (C4). PC786 PK was measured in plasma and in nasal mucosal lining fluid (MLF) collected using a synthetic absorptive matrix. Results PC786 was well tolerated, with no significant adverse clinical nor laboratory findings. Following single inhaled doses PC786 appeared rapidly in the plasma; mean plasma Cmax of 190, 571, 1,760, and 3,270 pg/mL, for the 0.5, 2, 8, and 20 mg doses, respectively, were measured on average at 0.68 to 0.93 hours (Tmax) post-inhalation. Following administration of 5 mg BD (C2) the extent of accumulation was approximately 2-fold. The geometric mean apparent terminal half-life measured following 10 mg BD (C3) was 97 hours. The ratio of MLF:plasma concentrations ranged from 6,347 (+2 hours) to 1,050 (+24h). Conclusion PC786 was well tolerated by HVs and asthmatics. The compound showed a rapid Tmax, suggesting rapid exposure of the respiratory epithelium. The PC786 concentrations in MLF exceed the IC90 for RSV, but circulating plasma concentrations were low. The MLF:plasma measured in this study was consistent with lung:plasma ratios measured in preclinical studies. The long plasma half-life is consistent with slow absorption from the lung being the dominant process controlling systemic kinetic behavior. The long t½ and 2-fold accumulation ratio observed on repeat dosing supports once daily dosing in subsequent studies. Disclosures L. Cass, Pulmocide Ltd.: Employee and Shareholder, Salary. A. Davis, Pulmocide Ltd.: Employee and Shareholder, Salary. A. Murray, Pulmocide Ltd.: Employee and Shareholder, Salary. K. Woodward, Pulmocide Ltd.: Consultant, Consulting fee. K. Ito, Pulmocide Ltd.: Employee and Shareholder, Salary. P. Strong, Pulmocide Ltd.: Board Member, Employee and Shareholder, Salary. G. Rapeport, Pulmocide Ltd.: Board Member, Employee and Shareholder, Salary.

Background: Plasminogen activator inhibitor-1 (PAI-1) is an important regulator of fiat sites of vascular injury and thrombus formation. Recently, sputum PAI-1 was reported to be elevated in COPD. However, the mechanism of PAI-1 elevation in COPD has yet to be clarifi ed. Here, we show that PAI-1 elevation in COPD is closely associated with oxidative stress-induced nuclear factor k B (NF- k B) activation. Methods : Patients and control subjects were recruited from the outpatient department of Royal Brompton Hospital, local general practice, and the National Heart and Lung Institute. Sputum samples were obtained, and sputum sample processing was performed to obtain sputum supernatants and sputum macrophages. Results: The mean PAI-1 level in COPD sputum (1.92 3.11 ng/mL, n 5 32) was higher than that of both age-matched smokers without COPD (0.48 0.63 ng/mL, n 5 11) and healthy nonsmokers (0.55 1.11 ng/mL, n 5 9). Sputum PAI-1 signifi cantly correlated with sputum malondialdehyde (MDA) in COPD ( r 5 0.59, P , .001). In addition, NF- k B activity in sputum macrophages (three control and seven COPD subjects) signifi cantly correlated with both sputum PAI-1 ( r 5 0.72, P , .05) and sputum MDA ( r 5 0.78, P , .01). An in vitro study showed that both hydrogen peroxide and cigarette smoke-conditioned medium induced PAI-1 production in A549 cells, and the production was inhibited by an inhibitor of I k B kinase- b in a concentration-dependent manner. Furthermore, histone deacetylase 2 (HDAC2) knockdown by RNA interference, a mimic of oxidative-stress-dependent HDAC2 reduction, enhanced tumor necrosis factor- a -induced PAI-1 induction (half maximal effective concentration [EC 50 ], 0.64 0.19 ng/mL in HDAC2-KD, 7.64 3.70 ng/mL in control) concomitant with enhancement of NF- k B p65 acetylation and NF- k B DNA-binding activity. Conclusions: Oxidative stress, directly or indirectly via HDAC reduction, plays a role in PAI-1 expression in COPD via activation of NF- k B.

Chronic obstructive pulmonary disease (COPD) has been recently characterized as a disease of accelerated lung aging. The prevalence of COPD is age-dependent suggesting an intimate relationship between the pathogenesis of COPD and aging. Lung function decline, the hallmark feature of COPD evolution, is more prominent with increasing age and this decline is greater in smoking individuals. One of the major goals of COPD pharmacotherapy is the development of drugs that would be able to result in a decrease of the decline in lung function over years. However, till nowadays smoking cessation is the only known intervention which is able to decelerate lung function decline. Several mechanisms of aging, including oxidative stress, inflammation and telomere shortening have been shown to be implicated in COPD. Furthermore, numerous anti-aging molecules, including sirtuins and Nrf-2 are reduced, and pathways such as mTOR and genes such as Klotho have also been shown to be abnormal in the lungs of COPD patients. The above mechanisms have been associated with the accelerated lung aging in COPD patients. Numerous therapeutic interventions have been studied in an attempt to reverse accelerated lung aging, and some of them have already been tested in clinical trials. The aim of the present review is to summarize the mechanisms associated with the accelerated lung aging in COPD and to provide information about the possible therapeutic implications targeting those mechanisms.

Corticosteroid insensitive inflammation is one of the important hallmarks of chronic obstructive pulmonary disease (COPD). Tobacco smoke (TS) and poly(I:C) reduce corticosteroid responsiveness by down-regulation of histone deacetylase 2 (HDAC2). Previous in vitro study suggests that TNF-α also reduces corticosteroid responsiveness. Therefore, we examined the effects of the combination of TNF-α neutralization and dexamethasone (DEX) treatment on corticosteroid insensitive inflammation. Mice were exposed to TS for 11 days, followed by administration of poly(I:C) and DEX for 3 days, and also treated with anti-TNF-α antibody for 14 days once every other day. Mice were sacrificed 24 hours after the latest poly(I:C) and DEX administration, and bronchoalveolar lavage fluid (BALF) and lungs were collected. Inflammatory cells in BALF were measured by flow cytometry. The mRNA expression levels in lungs were measured by real-time PCR method. TS and poly(I:C) induced marked accumulation of neutrophils in BALF and enhancement of TNF-α mRNA expression in lungs. Furthermore, TS and poly(I:C) also decreased in HDAC2 mRNA levels in lungs. The combination of anti-TNF-α antibody and DEX treatment suppressed the increases in neutrophils and TNF-α mRNA levels, even though anti-TNF-α antibody alone had no effects on these increases. In addition, the combination of anti-TNF-α antibody and DEX treatment also restored HDAC2 mRNA levels. These results suggest that TNF-α will contribute to the decrease of corticosteroid responsiveness and the combination of TNF-α neutralization and corticosteroid treatment will provide a new therapeutic approach of COPD.

Introduction and background. The protein deacetylase sirtuin-1 (SIRT1) is and anti-ageing molecule that is decreased in the lung from patients with chronic obstructive pulmonary disease (COPD). Recently, SIRT1 has been detected in the serum and is increased in the asthmatic patients. Aims and objectives. To determine serum SIRT1 levels in the patients with COPD as a potential disease biomarker. Methods. Serum SIRT1 was measured by Western blotting, and relative ratio of band density in samples against that of positive control were calculated. Results. Several SIRT1 bands with differing molecular weights, including a 75kDa truncated form and 120kDa (actual size) were observed by Western blotting. The 120kDa serum SIRT1 was significantly decreased in the patients with COPD compared to the subjects without COPD (SIRT1 ratio in healthy: 0.90±0.34, vs COPD: 0.68±0.24; p=0.014), and positively correlated with the lung function (FEV 1 /FVC; r=0.31; p=0.020) and severity of airflow obstruction represented by FEV 1 % predicted (r=0.29; p=0.029). The 120kDa serum SIRT1 also showed; 1) positive correlation with BMI (r=0.36; p=0.0078) and diffusing capacity of the lung per unit volume (KCO%; r=0.32; p=0.025); 2) the tendency to decrease in the presence of lung emphysematous change (without emphysema: 0.92±0.37 vs with emphysema: 0.71±0.24; p=0.026) and clinical history of frequent COPD exacerbations (infrequent: 0.76±0.20 vs frequent: 0.56±0.26; p=0.027). No reduction was seen in 75KDa SIRT1. Conclusions. Serum SIRT1 was decreased in the patients with COPD, and might be a potential biomarkers of COPD and particularly accelerated ageing.

Inflammation, which is induced and perpetuated by microorganism pathogens or from the damage or death of host cells, is an essential component of various common respiratory diseases, such as chronic obstructive pulmonary disease (COPD), asthma, bronchiectasis, and acute respiratory distress syndrome (ARDS), as well as of less common ones, for example, sarcoidosis. Although different diseases express distinct inflammatory responses, there are some shared common features. For example, the Th2-related eosinophilic inflammation, which is typically present in asthma, is also characteristic of eosinophilic phenotype of COPD. On the other hand, the innate neutrophilic inflammation shared by a number of diseases, including COPD and ARDS, could be demonstrated in some patients with severe asthma. In this special issue, common inflammation was dissected in diverse ways by the authors to better understand the related respiratory diseases, from the clinical to the underlying mechanisms, pointing to future therapeutic prospect. Innate immunity plays a primary role in host defense against microorganism pathogens and in response to the damage or death of host cells; however, when dysregulated, instead, it changes to an essential component of various respiratory diseases. In this issue, neutrophilic inflammation was demonstrated to be involved in two clinical conditions poorly understood. F. Tang et al. investigated the neutrophilic inflammation in patients with suboptimally controlled asthma and observed a defective innate immunity, which might be linked to pathogen-induced exacerbations. F. L. Dente et al. suggested sputum neutrophilic inflammation as a good biomarker for disease severity of stable noncystic fibrosis bronchiectasis by careful investigation of correlation with clinical parameters in those subjects. K. Baines et al. unearth the role of the innate immune antimicrobial peptide β-defensin-1 in COPD and severe asthma, which is involved in persistent inflammation in those conditions and they also suggested it as a potential biomarker in sputum and a therapeutic target. Using similar approach, A. K. Barton et al. correlated BAL MMPs/TIMPs with clinical and cytological findings in different equine chronic pneumopathies and suggested that their ratios can be a good biomarker for disease severity and used for identifying subclinical cases. A number of the articles addressed the regulatory mechanisms or novel therapies for ARDS, a disease with overwhelming nonspecific inflammation evoked by variable etiology. L. Ma et al. tested the protective roles of 3,5,4′-tri-O-acetylresveratrol (AC-Rsv), a prodrug of resveratrol, in LPS-induced ARDS, focusing on the modulation of SIRT1 and the mitogen-activated protein kinase (MAPK) pathway. In a review article, Z. Xu et al. on the other hand gave attention to the dark side of histones, the extracellular histones, acting as new members of damage-associated molecular pattern (DAMP) molecules. C. F. Goncalves-de-Albuquerque et al. extensively reviewed the oleic acid-induced lung injury and ARDS. In addition to discussing the featured pathophysiological changes and the underlying pathogenesis, they also shed light on a link between lipid metabolism and inflammatory diseases. Finally, W.-C. Chou et al. reported a therapeutic role of caffeine in mitigating acute lung injury induced by ischemia-reperfusion of lower limbs. To combat the pathogens evading the innate response, the immune system mounts adaptive immunity. The immune response to tuberculosis is such a prototype, for example, Th1 immune response. However, in the absence of known etiology, granulomatous inflammation characterized with infiltration of activated Th1/Th17 lymphocytes as well as macrophages features the pathological changes of pulmonary sarcoidosis. By using correlation network analysis into BAL cells in sarcoidosis, T. Dyskova et al. addressed the contribution of both microRNAs and the Th1-transcription factor T-bet to the regulation of cytokine/chemokine-receptor network, which drives the inflammatory granulomatous disease. Thus, inflammatory pulmonary diseases, as shown in this special issue, usually recapture the inflammation evoked by the immune system to the invading pathogens or other environmental stress. Thus, one mechanism might underlie variable diseases, directing to a common therapy. Kang-Yun Lee Kazuhiro Ito Kittipong Maneechotesuwan

A 44 year old male presented with gait disturbance and showed rigidity, bradykinesia and small steppage gait. In his infancy, he had mental and motor retardation. MRI revealed pathognomonic iron deposition. He was diagnosed to have β-propeller protein-associated neurodegeneration (BPAN) with a de novo somatic mosaic mutation in WDR45.

BACKGROUND AND PURPOSE Patients with chronic obstructive pulmonary disease (COPD) show a poor response to corticosteroids, which has been linked to oxidative stress. Here we show that the long-acting β2-agonist formoterol (FM) reversed corticosteroid insensitivity under oxidative stress via inhibition of phosphoinositide-3-kinase (PI3K) signalling.

Chronic obstructive pulmonary disease (COPD) progresses very slowly and the majority of patients are therefore elderly. COPD is characterized by an abnormal persistent inflammatory response to noxious environmental stimuli and there are increasing evidences for a close relationship between premature aging and chronic inflammatory diseases. Thus, COPD is considered to be a disease of an accelerating aging. In this review, we collected the evidence for roles of aging on pathogenesis of COPD and considered future therapeutic strategy for COPD based on this senescence hypothesis. Since calorie restriction has been proved to extend lifespan, many efforts were made to clarify the molecular mechanism of aging. Aging is defined as the progressive decline of homeostasis that occurs after the reproductive phase of life is complete, leading to an increasing risk of disease or death due to impaired DNA repair after damage by oxidative stress or telomere shortening as a result of repeated cell division. During aging, pulmonary function progressively deteriorates; innate immunity is impaired and pulmonary inflammation increases, accompanied by structural changes, such as an enlargement of airspaces. Noxious environmental gases, such as cigarette smoke, may worsen these aging-related events in the lung or accelerate aging of the lung due to reduction in anti-aging molecules and/or stimulation of aging molecules. Aging signaling are complex but conserved in divert species, such as worm, fruit fry, rodent and humans. Especially the insulin like growth factor (IGF-1) signaling was well documented. Geroprotectors are therapeutics that affect the root cause of aging and age-related diseases, and thus prolong the life-span of animals. Most of geroprotectors such as melatonin, metformin, rapamycin and resveratrol are anti-oxidant or anti-aging molecule regulators. Therefore, geroprotection for the lung might be an attractive approach for the treatment of COPD by preventing premature aging of lung.

Background: Hypoxia inducible factor (HIF)-1 plays an important role in cellular adaptation to hypoxia by activating oxygen-regulated genes such as vascular endothelial growth factor (VEGF) and erythropoietin. Sputum VEGF levels are reported to be decreased in COPD, despite hypoxia. Here we show that patients with COPD fail to induce HIF-1α and VEGF under hypoxic condition because of a reduction in histone deacetylase (HDAC) 7. Methods: Peripheral blood mononuclear cells (PBMCs) were obtained from patients with moderate to severe COPD (n = 21), smokers without COPD (n = 12), and nonsmokers (n = 15). PBMCs were exposed to hypoxia (1% oxygen, 5% CO2, and 94% N2) for 24 h, and HIF-1α and HDAC7 protein expression in nuclear extracts were determined by sodium dodecyl sulfate poly acrylamide gel electrophoresis (SDS-PAGE)/Western blotting. Results: HIF-1α was significantly induced by hypoxia in each group when compared with the normoxic condition (12-fold induction in nonsmokers, 24-fold induction in smokers without COPD, fourfold induction in COPD), but induction of HIF-1α under hypoxia was significantly lower in patients with COPD than in nonsmokers and smokers without COPD (P < .05 and P < .01, respectively). VEGF messenger RNA detected by quantitative real-time polymerase chain reaction was correlated with HIF-1α protein in nuclei (r = 0.79, P < .05), and HDAC7 protein expression was correlated with HIF-1α protein in nuclei (r = 0.46, P < .05). HDAC7 knockdown inhibited hypoxia-induced HIF-1α activity in U937 cells, and HIF-1α nuclear translocation and HIF-1α binding to the VEGF promoter in A549 cells. Conclusions: HDAC7 reduction in COPD causes a defect of HIF-1α induction response to hypoxia with impaired VEGF gene expression. This poor cellular adaptation might play a role in the pathogenesis of COPD.

SummaryBackgroundCombination therapy with inhaled corticosteroids (ICS) and long-acting β2-adrenergic agonists (LABA) is reported to have superior effects on controlling asthma symptoms to ICS alone; however, there is no molecular-based evidence to explain the clinical effects. Here, the effect of the ICS/LABA combination was compared with ICS on glucocorticoid receptor (GR) activation in sputum macrophages.MethodsIn a randomised, double-blind cross-over placebo-controlled 6-visit study, 10 patients with mild asthma were given placebo, formoterol (Oxis® 12 μg), budesonide (Pulmicort® 200 μg :BUD200, or 800 μg :BUD800), or budesonide/formoterol combination (Symbicort®) as a single 100/6 μg (SYM100) or double 200/12 μg (SYM200) dose. Sputum macrophages were separated by plate adhesion from induced sputum. GR binding to the glucocorticoid-response elements on oligonucleotides (GR-GRE binding) was evaluated by ELISA. mRNA expression of MAP-kinase phosphatase (MKP)-1 and IL-8 were measured by quantitative RT-PCR.ResultsGR-GRE binding was significantly increased after treatment with SYM100 (3.5 OD/10 μg protein, median, p

Corticosteroid insensitivity (CI) is a major barrier to treating severe asthma. Despite intensive research, the molecular mechanism of CI remains uncertain. The aim of this study was to determine abnormality in corticosteroid action in severe asthma and to identify the molecular mechanism of the long-acting β(2)-adrenergic agonists (LABAs) formoterol and salmeterol on restoration of corticosteroid sensitivity in severe asthma in vitro. Peripheral blood mononuclear cells (PBMCs) were obtained from 16 subjects with severe corticosteroid-insensitive asthma, 6 subjects with mild corticosteroid-sensitive asthma, and 11 healthy volunteers. Corticosteroid (dexamethasone) sensitivity was determined on tumor necrosis factor-α (TNF-α)-induced interleukin (IL)-8 production. Glucocorticoid receptor (GR) phosphorylation and kinase phosphorylation were evaluated by immunoprecipitation-Western blotting analysis and kinase phosphorylation array in IL-2/IL-4-treated corticosteroid insensitive model in PBMCs. In vitro corticosteroid sensitivity on TNF-α-induced IL-8 production was significantly lower in patients with severe asthma than in healthy volunteers and patients with mild asthma. This CI seen in severe asthma was associated with reduced GR nuclear translocation and with hyperphosphorylation of GR, which were reversed by LABAs. In IL-2/IL-4-treated PBMCs, LABAs inhibited phosphorylation of Jun-NH(2)-terminal kinase and p38 mitogen-activated protein kinase-γ (p38MAPK-γ) as well as GR. In addition, cells with p38MAPK-γ knockdown by RNA interference did not develop CI in the presence of IL-2/IL-4. Furthermore, p38MAPK-γ protein expression was up-regulated in PBMCs from some patients with severe asthma. In conclusion, p38 MAPK-γ activation impairs corticosteroid action and p38 MAPK-γ inhibition by LABAs has potential for the treatment of severe asthma.

COPD is characterized by chronic airflow limitation and inflammation of the respiratory tract. Several inflammatory biomarkers have been evaluated in COPD but are poorly related to disease severity and progression. Osteoprotegerin (OPG) is a glycoprotein mediator that is expressed in the lung and macrophages, so we have studied its concentration in induced sputum and macrophages of patients with COPD.OPG was measured by enzyme-linked immunosorbent assay in induced sputum of patients with COPD and control subjects.OPG concentrations in induced sputum of patients with COPD (18.7 ± 18.6 ng/mL, n = 39) were significantly higher than those of healthy smokers (8.1 ± 5.6 ng/mL, n = 15), healthy nonsmokers (3.5 ± 3.8 ng/mL, n = 14), or patients with asthma (8.0 ± 5.4 ng/mL, n = 18). Sputum OPG levels in COPD negatively correlated with FEV(1) and positively correlated with residual volume to total lung capacity ratio (RV/TLC) (r = 0.55, P.05), transfer factor of the lung for carbon monoxide (r = -0.53, P.05), and carbon monoxide transfer coefficient (r = -0.61, P.01). By contrast, sputum IL-8 concentrations were related to disease severity but not to RV/TLC or gas diffusion. Airway macrophages and neutrophils were positive for OPG by immunocytochemistry in sputum and peripheral lung tissue. OPG induced matrix metalloproteinase-9 release from sputum macrophages in vitro.Sputum OPG may be a useful biomarker to monitor parenchymal destruction in COPD.

Chronic obstructive pulmonary disease (COPD) is characterised by oxidative stress and increased risk of lung carcinoma. Oxidative stress causes DNA damage which can be repaired by DNA-dependent protein kinase complex.To investigate DNA damage/repair balance and DNA-dependent protein kinase complex in COPD lung and in an animal model of smoking-induced lung damage and to evaluate the effects of oxidative stress on Ku expression and function in human bronchial epithelial cells.Protein expression was quantified using immunohistochemistry and/or western blotting. DNA damage/repair was measured using colorimetric assays.8-OH-dG, a marker of oxidant-induced DNA damage, was statistically significantly increased in the peripheral lung of smokers (with and without COPD) compared with non-smokers, while the number of apurinic/apyrimidinic (AP) sites (DNA damage and repair) was increased in smokers compared with non-smokers (p = 0.0012) and patients with COPD (p0.0148). Nuclear expression of Ku86, but not of DNA-PKcs, phospho-DNA-PKcs, Ku70 or γ-H2AFX, was reduced in bronchiolar epithelial cells from patients with COPD compared with normal smokers and non-smokers (p0.039). Loss of Ku86 expression was also observed in a smoking mouse model (p0.012) and prevented by antioxidants. Oxidants reduced (p0.0112) Ku86 expression in human bronchial epithelial cells and Ku86 knock down modified AP sites in response to oxidative stress.Ineffective DNA repair rather than strand breakage per se accounts for the reduced AP sites observed in COPD and this is correlated with a selective decrease of the expression of Ku86 in the bronchiolar epithelium. DNA damage/repair imbalance may contribute to increased risk of lung carcinoma in COPD.

Disseminated intravascular coagulation (DIC) is a life-threatening condition that frequently occurs in patients with hematologic malignancies. Currently, recombinant human soluble thrombomodulin (rTM) is a therapeutic DIC drug that is manufactured and sold in Japan only. We evaluated the efficacy of rTM compared to that of gabexate mesilate (GM), which was previously used routinely for treating DIC in Japan, in patients with acute myeloid leukemia (AML). This retrospective study enrolled 43 AML patients, including 17 with acute promyelocytic leukemia (APL), that was complicated with DIC. DIC resolution rates in non-APL AML and rTM-treated APL patients were 68.4% and 81.8%, respectively. In non-APL AML patients, the duration of rTM administration was significantly shorter than that of GM (7 vs 11 days), suggesting that rTM could improve DIC earlier than GM, although rTM was used in patients with more severe DIC. Moreover, treatment with rTM significantly improved DIC score, fibrinogen, fibrin/fibrinogen degradation product (FDP), and prothrombin time (PT) ratio. Conversely, treatment with GM only improved the DIC score and FDP. In APL patients, the duration of rTM administration was also significantly shorter than that of GM. No severe side effects associated with the progression of bleeding were observed during rTM administration. These findings suggest that rTM is safe, and its anti-DIC effects are more prompt than GM for treating AML patients with DIC.

Patients with chronic obstructive pulmonary disease (COPD) show a poor response to corticosteroids. This has been linked to a reduction of histone deacetylase-2 as a result of oxidative stress and is reversed by theophylline.To determine the role of phosphoinositide-3-kinase-delta (PI3K-δ) on the development of corticosteroid insensitivity in COPD and under oxidative stress, and as a target for theophylline.Corticosteroid sensitivity was determined as the 50% inhibitory concentration of dexamethasone on tumor necrosis factor-α-induced interleukin-8 release in peripheral blood mononuclear cells from patients with COPD (n = 17) and compared with that of nonsmoking (n = 8) and smoking (n = 7) control subjects. The effect of theophylline and a selective PI3K-δ inhibitor (IC87114) on restoration of corticosteroid sensitivity was confirmed in cigarette smoke-exposed mice.Peripheral blood mononuclear cells of COPD (50% inhibitory concentration of dexamethasone: 156.8 ± 32.6 nM) were less corticosteroid sensitive than those of nonsmoking (41.2 ± 10.5 nM; P = 0.018) and smoking control subjects (47.5 ± 19.6 nM; P = 0.031). Corticosteroid insensitivity and reduced histone deacetylase-2 activity after oxidative stress were reversed by a non-selective PI3K inhibitor (LY294002) and low concentrations of theophylline. Theophylline was a potent selective inhibitor of oxidant-activated PI3K-δ, which was up-regulated in peripheral lung tissue of patients with COPD. Furthermore, cells with knock-down of PI3K-δ failed to develop corticosteroid insensitivity with oxidative stress. Both theophylline and IC87114, combined with dexamethasone, inhibited corticosteroid-insensitive lung inflammation in cigarette-smoke-exposed mice in vivo.Inhibition of oxidative stress dependent PI3K-δ activation by a selective inhibitor or theophylline provides a novel approach to reversing corticosteroid insensitivity in COPD.

Nitric oxide (NO) is increased in the lung periphery of patients with chronic obstructive pulmonary disease (COPD). However, expression of the NO synthase(s) responsible for elevated NO has not been identified in the peripheral lung tissue of patients with COPD of varying severity.Protein and mRNA expression of nitric oxide synthase type I (neuronal NOS [nNOS]), type II (inducible NOS [iNOS]), and type III (endothelial NOS [eNOS]) were quantified by Western blotting and reverse transcription-polymerase chain reaction, respectively, in specimens of surgically resected lung tissue from nonsmoker control subjects, patients with COPD of varying severity, and smokers without COPD, and in a lung epithelial cell line (A549). The effects of nitrative/oxidative stress on NOS expression and activity were also evaluated in vitro in A549 cells. nNOS nitration was quantified by immunoprecipitation and dimerization of nNOS was detected by low-temperature SDS-PAGE/Western blot in the presence of the peroxynitrite generator, 3-morpholinosydnonimine-N-ethylcarbamide (SIN1), in vitro and in vivo.Lung tissue from patients with severe and very severe COPD had graded increases in nNOS (mRNA and protein) compared with nonsmokers and normal smokers. Hydrogen peroxide (H(2)O(2)) and SIN1 as well as the cytokine mixture (IFN-gamma, IL-1beta, and tumor necrosis factor-alpha) increased mRNA expression and activity of nNOS in A549 cells in a concentration-dependent manner compared with nontreated cells. Tyrosine nitration resulted in an increase in nNOS activity in vitro, but did not affect its dimerization.Patients with COPD have a significant increase in nNOS expression and activity that reflects the severity of the disease and may be secondary to oxidative stress.

Resumo: A DPOC constitui um problema de saúde global de importância crescente, com enorme impacto nos custos directos e indirectos em recursos de saúde. Apesar do seu impacto, pouco se sabe ainda sobre os mecanismos celulares, moleculares e genéticos desta doença, e as te-rapêuticas farmacológicas actualmente disponíveis não influenciam a progressão da doença ou a mortalidade. A limitação do débito aéreo avaliada pela redução do volume expiratório máximo no 1.º segundo (FEV1) progride muito lentamente ao longo de várias décadas, condicionando o aparecimento de sintomas em adultos acima dos 40 anos ou já na terceira idade. Desta forma, a prevalência da DPOC é dependente da idade, sugerindo uma relação íntima entre a pato-génese da DPOC e a do envelhecimento.A senescência ou processo de envelhecimento definese como o declínio progressivo da homeostasia que ocorre após estar completa a fase reprodutiva da vida e conduz a um risco aumentado de doença e de morte. Segundo Kirkwood1, o envelhecimento resulta da interacção entre a lesão e a reparação, como resultado da energia produzida pelo indivíduo para manter a integridade orgânica e proteger o ADN da agressão oxidativa. A falência orgânica ou celular na manutenção ou reparação resulta de uma acção integrada entre genes, ambiente e defeitos intrínsecos do organismo. Subjacente ao processo de envelhecimento, existe uma acumulação progressiva de danos a nível molecular.As alterações a nível celular causam reacções inflamatórias, e estas, por sua vez, exacerbam as lesões celulares existentes. Desta forma, os factores inflamatórios e anti-inflamatórios modulam a evolução do envelhe-cimento. As alterações inflamatórias e estruturais associadas ao envelhecimento resultam da falência em eliminar os radicais de oxigénio (ROS), da falência em reparar o ADN lesado e do encurtamento do teló-mero. Os telómeros protegem as extremidades dos cromossomas, mas, quando se encontram expostos a elevados níveis de stress oxidativo, vão-se encurtando progressivamente à medida que as células se dividem. Com o envelhecimento, a perda e encurtamento dos telómeros condicionam o declínio da capacidade para as células se dividirem â senescência replicativa.O stress oxidativo causa lesão do ADN, o que acelera o processo de envelhecimento e aumenta o risco de cancro (exemplo, a senescência da glândula mamária e o risco elevado de cancro da mama).Os gases ambientais, como o fumo do cigarro ou outros poluentes, podem acelerar o envelhecimento do pulmão ou agravar os eventos relacionados com o en-velhecimento pulmonar através de uma resolução defeituosa da inflamação. A redução das moléculas anti-envelhecimento como as histona desacetilases e as sirtuínas, pode igualmente induzir a progressão acelerada para a DPOC.Ainda não é claro como é que o processo de envelhe-cimento está envolvido no declínio da função pulmonar e na inflamação da DPOC. Contudo, o pulmão do idoso e o pulmão do doente com DPOC apresen-tam muitas semelhanças. A lesão provocada pelo fumo do cigarro e outros pneumopoluentes conduz a um declínio mais acelerado da função pulmonar, com falência dos mecanismos de reparação e de manuten-ção. A presença de inflamação nos dois processos (en-velhecimento e DPOC) traduz-se em acumulação de neutrófilos, na activação da NF-kB e na elevação dos níveis plasmáticos de interleucinas IL-6, IL-8 e TNF-α. Também os telómeros das células alveolares tipo II, das células endoteliais e das células mononu-cleares em doentes com enfisema são significativamente mais curtos do que em indivíduos não fumadores da mesma idade.Neste artigo, os autores descrevem ainda estudos recentes sobre os mecanismos de transdução de sinal, como as vias da acetilação das proteínas envolvidas no processo de envelhecimento, identificando assim novas moléculas anti-envelhecimento que poderão constituir abordagens inovadoras na terapêutica da DPOC. Os antioxidantes actualmente disponíveis, como a N-acetilcisteína, não são suficientemente potentes para reduzir o stress oxidativo nos pulmões. Existem vários fármacos em desenvolvimento, como os novos análogos da glutationa e da superoxido dismutase (exemplo, o sulforafano) e novas moléculas antienvelhecimento com maior controlo sobre a resis-tência ao stress oxidativo, a reparação do ADN e a inflamação, como os activadores das sirtuínas (exem-plo, o resveratrol, o activador específico da SIRT1 ou o activador da SIRT6).

Peroxynitrite (PN) formed by the reaction of nitric oxide and superoxide is a powerful oxidant/nitrosant. Nitrative stress is implicated in COPD pathogenesis, but PN has not been detected due to a short half-life (1 s) at physiologic condition. Instead, 3-nitrotyrosine has been measured as a footprint of PN release.PN was measured using oxidation of 2',7'-dichlorofluorescein (DCDHF) in exhaled breath condensate (EBC) collected in high pH and sputum cells. The PN scavenging effect was also evaluated by the same system as PN-induced bovine serum albumin (BSA) nitration.The mean (+/- SD) PN levels in EBC of COPD patients (7.9 +/- 3.0 nmol/L; n = 10) were significantly higher than those of healthy volunteers (2.0 +/- 1.1 nmol/L; p0.0001; n = 8) and smokers (2.8 +/- 0.9 nmol/L; p = 0.0017; n = 6). There was a good correlation between PN level and disease severity (FEV(1)) in COPD (p = 0.0016). Fudosteine (FDS), a unique mucolytic antioxidant, showed a stronger scavenging effect of PN than N-acetyl-cysteine on DCDHF oxidation in vitro and in sputum macrophages, and also on PN-induced BSA nitration. FDS (0.1 mmol/L) reduced PN-enhanced interleukin (IL)-1beta-induced IL-8 release and restored corticosteroid sensitivity defected by PN more potently than those induced by H(2)O(2) in A549 airway epithelial cells.This noninvasive PN measurement in EBC may be useful for monitoring airway nitrative stress in COPD. Furthermore, FDS has the potential to inhibit PN-induced events in lung by its scavenging effect.

Oligomer Abeta is the term utilized for multimeric but non-fibrillar forms of amyloid beta-protein (Abeta). The most prominent property of oligomer Abeta is considered to be its solubility and structure. Here, we examined the histochemical localization of oligomer Abeta in AD brains. At present, little information is available on the structure and function of cerebral oligomer Abeta. We therefore studied the molecular localization of oligomer Abeta using a newly generated polyclonal mouse antisera against a variant Abeta with a deletion mutation of the 22(nd) glutamate that we found recently in a patient with familial Alzheimer's disease. Intracellular as well as extracellular oligomer Abeta are herein discussed to define their structure and pathological roles in disease.

Background Nitric oxide (NO) is produced by resident and inflammatory cells in the respiratory tract by the enzyme NO synthase (NOS), which exists in three isoforms: neuronal NOS (nNOS), inducible NOS (iNOS), and endothelial NOS. NO production is increased in patients with COPD, and the production of NO under oxidative stress conditions generates reactive nitrogen species that may amplify the inflammatory response in COPD. Methods To examine the role of increased NO in COPD, we administered a relatively selective iNOS inhibitor, aminoguanidine, by nebulization in a double-blind, placebo-controlled study in COPD patients, healthy smokers, and healthy nonsmoking subjects. We investigated whether aminoguanidine had any effect on exhaled NO produced in the central lung (flux of NO from the airways [J no ] and peripheral lungs (concentration of NO in peripheral lung [C alv ], on NO metabolites (nitrite [NO 2 − ]/nitrate [NO 3 − ], peroxinitrite [ONOO − ], nitrotyrosine), and on a marker of oxidative stress (8-isoprostane) in exhaled breath condensate (EBC) and in sputum. Results Aminoguanidine administration resulted in a significant reduction in J no compared with administration of the saline solution control in healthy subjects, smokers, and COPD patients. C alv in smokers and in COPD patients was not completely inhibited 1 h after aminoguanidine inhalation, in marked contrast to previous results in asthma. Moreover, ONOO − and NO 2 − /NO 3 − levels were also increased in EBC and in sputum of smokers and COPD and were not completely inhibited following aminoguanidine inhalation. 8-Isoprostane levels were also increased in smokers and in COPD patients but were not reduced after aminoguanidine inhalation. Conclusions These results suggest that the constitutive NOS isoform as well as iNOS might be involved in NO release and contribute to the high C alv and ONOO − production in patients with COPD. Trial registration: Clinicaltrials.gov Identifier: NCT00180635.

There is increasing evidence for a close relationship between aging and chronic inflammatory diseases. COPD is a chronic inflammatory disease of the lungs, which progresses very slowly and the majority of patients are therefore elderly. We here review the evidence that accelerating aging of lung in response to oxidative stress is involved in the pathogenesis and progression of COPD, particularly emphysema. Aging is defined as the progressive decline of homeostasis that occurs after the reproductive phase of life is complete, leading to an increasing risk of disease or death. This results from a failure of organs to repair DNA damage by oxidative stress (nonprogrammed aging) and from telomere shortening as a result of repeated cell division (programmed aging). During aging, pulmonary function progressively deteriorates and pulmonary inflammation increases, accompanied by structural changes, which are described as senile emphysema. Environmental gases, such as cigarette smoke or other pollutants, may accelerate the aging of lung or worsen aging-related events in lung by defective resolution of inflammation, for example, by reducing antiaging molecules, such as histone deacetylases and sirtuins, and this consequently induces accelerated progression of COPD. Recent studies of the signal transduction mechanisms, such as protein acetylation pathways involved in aging, have identified novel antiaging molecules that may provide a new therapeutic approach to COPD.

Chronic obstructive pulmonary disease (COPD) patients have an increased number of T cells within their lungs. It is unknown whether these T cells, remain there forever or if there is a continuous turnover from the blood. In the adult, there is a significant T lymphocytopoiesis from the thymus producing cells known as recent thymic emigrants (RTEs). T cell receptor excision circles (TREC) are a marker of RTEs. We investigated the number of TREC in blood from patients with untreated stable, mild to moderate COPD (n=6) compared with age-matched smokers with normal lung function (n=6) and nonsmokers (n=8). The results showed variable expression of TREC in each subject group and no sig- nificant difference between TREC expressions in any group of subjects. Changes in T-cell numbers in the lung of stable COPD patients may reflect prolonged survival or proliferation of these cells within the lung rather than continuous re- cruitment from the blood.

Chronic obstructive pulmonary disease (COPD) is a chronic inflammatory airway disease. It has become one of the commonest diseases worldwide and a major global healthcare problem. The disease progresses very slowly and, in contrast to asthma, the majority of affected patients are middle-aged or elderly. There is increasing evidence of a direct relationship between aging and an increased incidence of chronic inflammatory disease. Thus, in this review, the link between aging of the lung and the progression of COPD is discussed. Aging, or senescence, is defined as the progressive decline in homeostasis that occurs after the reproductive phase of life is completed, leading to an increasing risk of disease or death. This is the result of a failure of organ maintenance resulting from DNA damage, oxidative stress and telomere shortening. During aging, pulmonary function progressively deteriorates and inflammation of the lung increases, and these changes are accompanied by structural changes. Environmental stimul...

Statins have immunomodulatory effects that may be beneficial in the treatment of asthma. We studied the effects of atorvastatin alone and in combination with inhaled beclometasone (ICS) on a range of sputum supernatant mediator concentrations from samples of a clinical trial that reported asthma control score improvements after short-term treatment with atorvastatin (Braganza G et al, BMC Pulm Med 2011). 39 samples were analysed for concentrations of 35 mediators (Luminex or biochemical analyses) implicated in asthma pathogenesis and their relationship with control scores. No marker was found to be reduced by ICS alone. Markers of inflammation, neutrophil function and remodelling pathways were reduced by the treatments: CCL7, IL-12p70, sCD40L, FGF-2, CCL4, TGF-α, and MMP-8 by atorvastatin as compared to placebo; MMP-8, IL-1β, IL-10, MMP-9, sCD40L, FGF-2, IL-7, G-CSF and CCL7 by atorvastatin + ICS as compared to ICS alone. Correlation analyses showed that a reduction in sputum neutrophil count was associated with a decrease in IL-6 [0.639 (0.286, 0.839), p=0.001 ] and CXCL8 [0.541 (0.153, 0.784), p=0.007 ] after atorvastatin treatment. After atorvastatin + ICS treatment, an improvement in ACQ score was associated with a reduction in sputum CCL2 [0.542 (0.165, 0.780), p=0.006 ] and CXCL8 [0.513 (0.126, 0.764), p=0.010 ], while an improvement in AQLQ score was associated with a reduction in CXCL8 [-0.572 (-0.797, -0.207), p=0.003 ], G-CSF [-0.543, (-0.781, 0.167), p=0.006 ] and IL-7 [-0.533 (-0.776, -0.154), p=0.007 ]. This combined data suggests that atorvastatin may affect markers of neutrophil function and airway remodelling in chronic asthma. Further long-term clinical studies are needed to elucidate these effects.

Background: Cognitive abilities strongly influence medication adherence among elderly individuals. We aimed to evaluate the relationship between medication adherence and cognitive decline using Lawton’s instrumental activities of daily living (IADL) scoring system and the Mini-Mental State Examination (MMSE). Methods: Receiver–operating characteristic (ROC) curves were used to evaluate the IADL scores and MMSE results. Results: The ROC curve analysis of the IADL and MMSE results revealed that the shopping (MMSE cutoff = 22 points, sensitivity = 0.726, and specificity = 0.683) and responsibility for own medications (MMSE cutoff = 22 points, sensitivity = 0.759, and specificity = 0.720) categories were associated with declining IADL scores during early stage cognitive dysfunction. Conclusion: Declining IADL scores in the shopping and responsibility for own medications categories may be effective indices for predicting early-stage cognitive dysfunction in elderly individuals. Cognitive dysfunction screening at pharmacy counters may be useful.

The phosphoinositide 3-kinase(s) (PI3K) are a family of proteins that catalyze the phosphorylation of the 3-OH position of phosphoinositides and generate lipids that control a wide variety of intracellular signaling pathways. They are classified into three families according to their structure and substrate specificity and are thought to have distinct biological roles. Recent studies suggested that numerous components of the PI3K pathway play a crucial role in the expression and activation of inflammatory mediators, inflammatory cell recruitment, immune cell function, airway remodeling, and corticosteroid insensitivity in chronic inflammatory respiratory disease. Selective PI3K inhibitors have been developed that reduce inflammation and some characteristics of disease in experimental animal models. Targeting specific PI3K isoforms that may be overexpressed or overactive in disease should allow for selective treatment of respiratory diseases. Encouraging data from animal models, primary cells and clinical studies in other diseases suggest that inhibitors of PI3K/Akt may prove to be useful novel therapies in the treatment of asthma and chronic obstructive pulmonary disease.