A microphysiological model of the bronchial airways reveals the interplay of mechanical and biochemical signals in bronchospasm.

In asthma, the contraction of the airway smooth muscle and the subsequent decrease in airflow involve a poorly understood set of mechanical and biochemical events. Organ-level and molecular-scale models of the airway are frequently based on purely mechanical or biochemical considerations and do not account for physiological mechanochemical couplings. Here, we present a microphysiological model of the airway that allows for the quantitative analysis of the interactions between mechanical and biochemical signals triggered by compressive stress on epithelial cells. We show that a mechanical stimulus mimicking a bronchospastic challenge triggers the marked contraction and delayed relaxation of airway smooth muscle, and that this is mediated by the discordant expression of cyclooxygenase genes in epithelial cells and regulated by the mechanosensor and transcriptional co-activator Yes-associated protein. A mathematical model of the intercellular feedback interactions recapitulates aspects of obstructive disease of the airways, which include pathognomonic features of severe difficult-to-treat asthma. The microphysiological model could be used to investigate the mechanisms of asthma pathogenesis and to develop therapeutic strategies that disrupt the positive feedback loop that leads to persistent airway constriction.

ß-Elemene inhibits the proliferation of primary human airway granulation fibroblasts by down-regulating canonical Wnt/ß-catenin pathway.

Benign airway stenosis is a clinical challenge because of recurrent granulation tissues. Our previous study proved that a Chinese drug, ß-elemene, could effectively inhibit the growth of fibroblasts cultured from hyperplastic human airway granulation tissues, which could slow down the progression of this disease. The purpose of the present study is to find out the mechanism for this effect. We cultured fibroblasts from normal human airway tissues and human airway granulation tissues. These cells were cultured with 160 µg/ml normal saline (NS), different doses of ß-elemene, or 10 ng/ml canonical Wnt/ß-catenin pathway inhibitor (Dickkopf-1, DKK-1). The proliferation rate of cells and the expression of six molecules involved in canonical Wnt/ß-catenin pathway, Wnt3a, glycogen synthase kinase-3ß (GSK-3ß), ß-catenin, α-smooth muscle actin (α-SMA), transforming growth factor-ß (TGF-ß), and Collagen I (Col-I), were measured. At last, we used canonical Wnt/ß-catenin pathway activator (LiCl) to further ascertain the mechanism of ß-elemene. Canonical Wnt/ß-catenin pathway is activated in human airway granulation fibroblasts. ß-Elemene didn't affect normal human airway fibroblasts; however, it had a dose-responsive inhibitive effect on the proliferation and expression of Wnt3a, non-active GSK-3ß, ß-catenin, α-SMA, TGF-ß, and Col-I of human airway granulation fibroblasts. More importantly, it had the same effect on the expression and nuclear translocation of active ß-catenin. All these effects were similar to 10 ng/ml DKK-1 and could be attenuated by 10 mM LiCl. Thus, ß-elemene inhibits the proliferation of primary human airway granulation fibroblasts by down-regulating canonical Wnt/ß-catenin pathway. This pathway is possibly a promising target to treat benign tracheobronchial stenosis.

Mechanistic link between diesel exhaust particles and respiratory reflexes.

BACKGROUND: Diesel exhaust particles (DEPs) are a major component of particulate matter in Europe's largest cities, and epidemiologic evidence links exposure with respiratory symptoms and asthma exacerbations. Respiratory reflexes are responsible for symptoms and are regulated by vagal afferent nerves, which innervate the airway. It is not known how DEP exposure activates airway afferents to elicit symptoms, such as cough and bronchospasm. OBJECTIVE: We sought to identify the mechanisms involved in activation of airway sensory afferents by DEPs. METHODS: In this study we use in vitro and in vivo electrophysiologic techniques, including a unique model that assesses depolarization (a marker of sensory nerve activation) of human vagus. RESULTS: We demonstrate a direct interaction between DEP and airway C-fiber afferents. In anesthetized guinea pigs intratracheal administration of DEPs activated airway C-fibers. The organic extract (DEP-OE) and not the cleaned particles evoked depolarization of guinea pig and human vagus, and this was inhibited by a transient receptor potential ankyrin-1 antagonist and the antioxidant N-acetyl cysteine. Polycyclic aromatic hydrocarbons, major constituents of DEPs, were implicated in this process through activation of the aryl hydrocarbon receptor and subsequent mitochondrial reactive oxygen species production, which is known to activate transient receptor potential ankyrin-1 on nociceptive C-fibers. CONCLUSIONS: This study provides the first mechanistic insights into how exposure to urban air pollution leads to activation of guinea pig and human sensory nerves, which are responsible for respiratory symptoms. Mechanistic information will enable the development of appropriate therapeutic interventions and mitigation strategies for those susceptible subjects who are most at risk.

Epidemiology and risk factors of tooth loss among Iranian adults: findings from a large community-based study.

OBJECTIVES: To investigate the prevalence of tooth loss and different prosthetic rehabilitations among Iranian adults, as well as the potential determinants of tooth loss. METHODS: In a cross-sectional community-based study conducted among 8094 Iranian adults living in Isfahan province, a self-administered questionnaire was used to assess epidemiologic features of tooth loss. RESULTS: Thirty-two percent of subjects had all their teeth, 58.6% had lost less than 6, and 7.2% of participants had lost more than 6 teeth. One hundred and sixty-nine individuals (2.2%) were edentulous. Among participants, 2.3% had single jaw removable partial denture, 3.6% had complete removable denture in both jaws, and 4.6% had fixed prosthesis. Others reported no prosthetic rehabilitation (89.5%). In the age subgroup analysis (≤35 and >35 years old) tooth loss was more prevalent among men than women (OR = 2.8 and 1.9, resp., P < 0.01). Also, in both age groups, current and former smokers had higher levels of tooth loss than nonsmokers (P < 0.001 and P < 0.05, resp.). In addition, tooth loss was positively related to metabolic abnormality for age group >35 years (adjusted OR = 1.29, P < 0.01). CONCLUSIONS: Tooth loss is highly prevalent in Iranian adult population. Community programs promoting oral health for prevention of tooth loss should be considered taking into account its major determinants including lower educational level, male gender, smoking, and metabolic abnormality.

IgE sensitization to the nonspecific lipid-transfer protein Ara h 9 and peanut-associated bronchospasm.

Allergen component analysis is now available in many laboratories. The aim of this study was to examine the possible association between peanut allergen IgE components and severity of clinical reactions in patients with a history of peanut allergy. Data and sera collected from 192 patients within the Manchester Allergy Research Database and Serum Bank were used in this retrospective study. Sensitization to peanut specific IgE and Ara h 1, 2, 3, and 8 peanut IgE components, as measured by fluoroenzyme immunoassay, was not associated with anaphylaxis. In contrast, sensitization to the lipid-transfer protein Ara h 9 was significantly more prevalent in patients with peanut-associated bronchospasm (26% versus 9% of patients), even after adjusting for potential confounding effects of age, gender, and severity of concomitant chronic atopic diseases. Patients who were sensitized to Ara h 9 were more likely to have ingested rather than just have had skin contact with peanut and have a more rapid onset of symptoms. These results are consistent with observations that sensitization to heat and protease resistant lipid-transfer protein components of hazelnut, grains, and fruit is predictive of anaphylaxis.

Lung function in patients with gastro-esophageal reflux disease and respiratory symptoms.

The aim of this study was to investigate lung function in patients with gastro-esophageal reflux disease (GERD) who present respiratory symptoms suggestive of the possibility of co-morbid asthma. The study encompassed 20 patients (9 women and 11 men; age range from 11 to 68 years) diagnosed with GERD and presenting with chronic cough and other non-specific periodic respiratory complaints. The control group consisted of closely gender and age-matched 20 subjects without any gastrointestinal or respiratory symptoms. All patients and control subjects were tested for lung function, which encompassed spirometric and flow-volume variables. We found that none of the GERD patients had lung function abnormalities characteristic of asthma. There were, however, decreases in forced expired volume in 1 s, forced vital capacity, and in maximal instantaneous forced expiratory flows in the GERD patients compared with the healthy subjects. We conclude that cough accompanying GERD is unlikely to be associated with the presence of co-morbid asthma, but rather suggests a mild airway inflammation developing as a sequel of GERD. The corollary is that chronic cough should prompt physician's attention to consider diagnostic work-up toward the possibility of GERD.

Design and synthesis of new 8-anilide theophylline derivatives as bronchodilators and antibacterial agents.

Theophylline derivatives have long been recognized as potent bronchodilators for the relief of acute asthma. Recently, it was found that bacterial infection has a role in asthma pathogenesis. The present work involves the design and synthesis of 8-substituted theophylline derivatives as bronchodilators and antibacterial agents. The chemical structures of these compounds were elucidated by IR, (1)H-NMR, mass spectrometry, and elemental analyses. The bronchodilator activity was evaluated using acetylcholine-induced bronchospasm in guinea pigs, and most of the compounds showed significant anti-bronchoconstrictive activity in comparison with standard aminophylline. In addition, the antibacterial activity of all the target compounds was investigated in vitro against Gram-positive and Gram-negative bacteria using ampicillin as a reference drug. Results showed that some of the tested compounds possessed significant antibacterial activity. A pharmacophore model was computed to obtain useful insight into the essential structural features of bronchodilator activity. A structure activity relationship was also discussed.

Effects of cigarette smoke extract and nicotine on bronchial tone and acetylcholine-induced airway contraction in mouse lung slices.

BACKGROUND: Tobacco smoke is a key risk factor for chronic obstructive pulmonary disease, but it may also alter the pathophysiology of asthma. In the present study, we analyzed whether tobacco smoke has acute or chronic effects on bronchial tone and whether it alters bronchial reactivity in vitro. METHODS: Airways in murine lung slices were digitally recorded and the change in cross-sectional area with time was quantified. T-bet KO mice served as a model for bronchial hyperreactivity. T-bet KO mice show a shift towards type 2 helper T lymphocytes and display histological as well as functional characteristics of asthma. Cigarette smoke extract (CSE) was obtained using commercially available cigarettes (Gauloise Blondes) by drawing cigarette smoke slowly through a water pump into a tube containing 10 mL of DMEM culture medium. RESULTS: Acute exposure to CSE led to relaxation of the airway. Acute exposure to nicotine resulted in a minor relaxation of the airway in Balb/C mice and in nonsignificant relaxation of the airway in T-bet KO mice. The nicotinic acetylcholine-receptor hexamethonium partially inhibited CSE-induced airway relaxation. Airway contraction in response to acetylcholine was stronger in T-bet KO mice than in Balb/C mice. After exposure to CSE or nicotine for 48 hours, acetylcholine-induced airway contraction was no longer different between the 2 types of mice. CONCLUSIONS: Our data indicate that acute exposure to CSE leads to airway relaxation, which is partially mediated by nicotine. Chronic exposure to CSE reverses bronchial hyperreactivity in the airways of T-bet KO mice; this effect can be mimicked by chronic exposure to nicotine.

[Response mechanisms of the airway smooth muscle tissue in experimental bronchial spasm].

This investigation was aimed at the complex evaluation of the reactivity mechanisms of bronchial smooth muscle tissue (SMT) in experimental bronchial spasm. Morphometric, cytospectrophotometric and electron microscopical analysis demonstrated the presence of three types of smooth muscle cells (SMC) within the bronchial SMT (small, medium, large), that differed in their linear and metabolic parameters. The findings of this study indicate that under the conditions of experimental bronchial spasm development, the ratios of SMC in bronchial SMT are changed with the increase in proportion of small SMC and the elimination of large SMC. In the dynamics of experimental bronchial spasm development, the activation of cytoplasmic synthesis as well as of DNA synthesis was detected mainly in group of small SMC. The reactive-dystrophic changes were marked at the subcellular level, that were most often identified in large SMC resulting in their elimination from population in the dynamics of an experiment. The data obtained suggest that one of the important mechanisms of airway SMT adaptation to the bronchial spasm development is a dynamic reorganization of SMC population.

Airway smooth muscle and bronchospasm: fluctuating, fluidizing, freezing.

We review here four recent findings that have altered in a fundamental way our understanding of airways smooth muscle (ASM), its dynamic responses to physiological loading, and their dominant mechanical role in bronchospasm. These findings highlight ASM remodeling processes that are innately out-of-equilibrium and dynamic, and bring to the forefront a striking intersection between topics in condensed matter physics and ASM cytoskeletal biology. By doing so, they place in a new light the role of enhanced ASM mass in airway hyper-responsiveness as well as in the failure of a deep inspiration to relax the asthmatic airway. These findings have established that (i) ASM length is equilibrated dynamically, not statically; (ii) ASM dynamics closely resemble physical features exhibited by so-called soft glassy materials; (iii) static force-length relationships fail to describe dynamically contracted ASM states; (iv) stretch fluidizes the ASM cytoskeleton. Taken together, these observations suggest that at the origin of the bronchodilatory effect of a deep inspiration, and its failure in asthma, may lie glassy dynamics of the ASM cell.

Evaluation of the effect of Myrica sapida on bronchoconstriction and bronchial hyperreactivity.

The present investigation was undertaken to evaluate the bronchodilator and bronchial hyperreactivity of the stem bark of Myrica sapida. Experimental models studied were histamine induced bronchospasm in guinea pigs, bronchoalveolar lavage fluid (BALF) in egg albumin sensitized guinea pigs, histamine release from the lung tissues of sensitized guinea pigs and histopathological studies. Ethanolic extract of M. sapida (75 mg/kg, p.o., for 7 days) showed significant protection against histamine aerosol induced bronchospasm. Significant decrease in the total and differential leukocyte counts in BALF and prevention of egg albumin induced histamine release from chopped lung tissues of sensitized guinea pigs was observed on chronic administration of ethanolic extract of M. sapida (75 mg/kg, p.o., for 15 days). Histological examination of the section of lung from sensitized guinea pigs treated with ethanolic extract of M. sapida (75 mg/kg, p.o., for 15 days) was comparable to that of the control group. These results suggest that M. sapida possesses not only bronchodilator activity but also decreases bronchial hyperresponsiveness by decreasing the infiltration of inflammatory mediators like eosinophils, neutrophils in BALF and inhibiting histamine release from lungs of sensitized guinea pigs.

Airway smooth muscle growth in asthma: proliferation, hypertrophy, and migration.

Increased airway smooth muscle mass is present in fatal and non-fatal asthma. However, little information is available regarding the cellular mechanism (i.e., hyperplasia vs. hypertrophy). Even less information exists regarding the functional consequences of airway smooth muscle remodeling. It would appear that increased airway smooth muscle mass would tend to increase airway narrowing and airflow obstruction. However, the precise effects of increased airway smooth muscle mass on airway narrowing are not known. This review will consider the evidence for airway smooth muscle cell proliferation and hypertrophy in asthma, potential functional effects, and biochemical mechanisms.

Time for a paradigm shift in asthma treatment: from relieving bronchospasm to controlling systemic inflammation.

Inflammation is a key pathology in asthma. In the central airways local inflammation leads to irreversible remodeling and airway dysfunction. Complex inflammatory changes also occur in the nose, sinuses, and small airways. In particular, rhinitis and asthma are linked by a common pathogenic process with common inflammatory cells, mediators, and cytokines. Cross-communication between the airways and bone marrow through inflammatory mediators in the circulation leads to systemic propagation of airway inflammation. Treatment of asthma has traditionally focused on relieving bronchospasm with beta(2)-agonists, which do not affect inflammation. Treatment of eosinophilic inflammation in the central airways with inhaled corticosteroids reduces local inflammation and improves pulmonary function but does not improve the systemic manifestations of asthma. If asthma is a systemic disease, the underlying systemic pathology should be targeted by identifying common disease mediators, mechanisms, or both that are triggered only during active disease. Of currently available therapies, leukotriene receptor antagonists block the action of cysteinyl leukotrienes and thus improve both asthma and rhinitis and other conditions systemically linked with asthma. Other potential treatments include receptor-blocking molecules and synthesis inhibitors related to eicosanoid inflammation. Treatment of asthma as a systemic disease requires clinical trials that evaluate the effects of new treatments on both lung function and the wider systemic pathology.

[Alternations of bronchial smooth muscle and effect of therapy on remodeling in asthma]. / Udzial miesniówki gladkiej w przebudowie oskrzeli oraz wplyw terapii na remodeling w astmie.

Alternations in airway wall architecture, particularly increased smooth muscle mass are associated with pathogenesis of asthma. Muscle fiber hyperplasia and hypertrophy is a major contributor to the increase in smooth muscle mass. Airway smooth muscle was traditionally considered to have only contractile and proliferative functions and has little attention with regard to its ability to express and release inflammatory mediators. Airway smooth muscle cells have been shown to release cytokines such as: GM-CSF, IL-11, IL-6, IL-1, IL-5, IL-8, PGs and NO. Airway remodeling has been shown to respond to some degree anti-inflammatory therapy. Several study results indicate that steroid can positively influence progressive airflow limitation. Combined use of a beta2-agonist and steroid can reduced the remodeling progression.

Airway remodeling from bench to bedside: current perspectives.

Bronchospasm and airway inflammation can lead to a constellation of irreversible changes in airway structure termed remodeling. Remodeling theory offers insight into the permanent biomechanical and pathologic alterations of asthmatic airways. Structural changes seen in asthmatic patients can include thickening of the airway wall reticular basement membrane (RBM), the presence of an abnormal elastic fiber network, and alterations in airway cartilage structure. Although steroid therapy is helpful in symptomatic control, it does not remedy structural alterations or many aspects of the inflammatory milieu. This article discusses several studies and supports the need for further investigation.

Pathological study of bronchospasms/tracheomalasia in patients with severe motor and intellectual disabilities.

This report concerns two autopsy cases of severe motor and intellectual disabilities (SMID) who died of bronchospasms or tracheomalasia. One case had no anatomical change in the tracheal wall except for an endotracheal granuloma, while the other showed softening of the tracheal wall. Since patients with SMID have risk factors for bronchospasms and tracheomalasia, such as gastro-esophageal reflux, aspiration, and thoracic deformities, it is important that we suspect the possibility of these conditions, when we see the respiratory distress in cases of SMID.

Effects of recycled paper dust extracts on isolated guinea pig trachea.

The effect of paper dust collected at two different locations in a paper recycling plant (PD1 and PD2) on isolated nonsensitized guinea pig tracheal smooth muscle was studied in vitro. Dust extracts were prepared as a 1:10 w/v aqueous solution. Dose-related contractions of guinea pig tracheal rings were elicited with both PD1 and PD2. Pharmacologic studies were performed with atropine (10(-6) M), indometacin (10(-6) M), pyrilamine (10(-6) M), LY171883 (10(-5) M), nordihydroguaiaretic acid (10(-5) M), and TMB8 (10(-5) M). The possible role of endogenous neuropeptides in this constrictor process was studied by depleting neural mediators with capsaicin (5 x 10(-6) M) before challenge with dust extracts. Constrictor effects were partially inhibited by a wide variety of the mediator blocking agents. The effects of both extracts were almost totally inhibited by the anticholinergic agent atropine, suggesting that a principal pathway mediating this response may involve the parasympathetic nervous system. The intracellular calcium-blocking agent TMB8 also induced a reduction of the contractile responses to PD1 and PD2 consistent with the well established role of intracellular calcium in smooth muscle constriction. Pretreatment with capsaicin significantly increased the contractile activity of paper dust extracts but only at the higher doses of these extracts. This suggests that the effect of paper dust is not initiated by the release of mediators stored in sensory nerves but that the prerelease of these mediators may enhance the constrictor effects of these dusts. We suggest that paper dust extracts cause dose-related airway smooth muscle constriction possibly associated with the release of cholinergic as well as other mediators. The constrictor effect does not require tissue presensitization or the release of neuropeptides from sensory nerves.