Investigating lung remodeling in Modul8-treated BALB/c asthmatic animals

Animal models of bronchial hyperresponsiveness have been successfully used to investigate the pathophysiology of asthma. When mice are sensitized and challenged with an allergen, such as OVA, they experience symptoms and processes similar to that of humans, and are therefore widely used as asthmatic animal models. In the current study the BALB/c murine asthmatic animal model was used to investigate the histological and ultrastructural changes that occur in the lungs of asthmatic animals that received no treatment, compared to two groups of asthmatic animals that were treated with a homeopathic immunodulator Modul8^® and hydrocortisone as positive control, respectively. Eosinophil counts in the bronchial lavage of the animals were also analyzed, since it is known that eosinophil counts are increased in the bronchial lavage in asthma. Results indicated that eosinophil counts were elevated in asthmatic animals compared to the controls, but were found to be significantly decreased in the treatment groups. Also, in the asthmatic, untreated animals, histological and ultrastructural changes, typically associated with the inflammatory process were found. Both treatment groups compared well to that of the control animals, indicating that the homeopathic product might be successfully used in the treatment of asthma.     

Voice problems as side effects of inhaled corticosteroids in asthma patients—a prevalence study

Voice disturbances in asthma patients may be caused by inhaled corticosteroids. In order to study the prevalence of such voice disturbances, a questionnaire was delivered to asthma patients at three asthma and allergy departments in Stockholm. The questionnaire consisted of 25 questions concerning the asthma disease symptoms, medication, voice function, and voice disturbances. A total of 350 questionnaires were delivered. Response frequency was 80%. There was a significant positive correlation between inhalation of cortison and voice disturbances. Most of the patients complained about hoarseness, followed by throat clearing, a lump in the throat, loss of voice, and less frequently, throat pain. There were no significantly differences between men and women. Elderly had more voice problems than young persons. Patients with voice-demanding professions had more problems than patients who used their voice to a lesser extent during the working day. There was a significant positive correlation between high cortison doses and voice problems as well as between voice problems and acid regurgitation.     

Airway reactivity-induced asthma in singers (Arias)

Professional singers are particularly attuned to changes in air flow that can alter their ability to perform. We describe four singers who complained primarily of performance-related vocalization problems, but who had normal larynges and fundamentally good voice technique. All had normal or near-normal spirometry. Subsequent studies revealed evidence of increased airway reactivity, either by response to bronchodilators or to methacholine. All responded to treatment with bronchodilators with improvement in vocal performance. This represents a form of airway reactivity-induced asthma in singers similar to that found with exercise or hyperventilation.     

Prokaryotic Expression, Purification, Antibody Production of a NH2 -Terminal Fragment of mCLCA3 Protein and Analysis of mClCA3 Protein Expression in Asthmatic Mouse Lung

mCLCA3 is a member of calcium activated chloride channel(CACC)family that may play an important role in mucin packaging and secretion in asthmatic and cystic fibrosis lung.To study the protein structure and expression of mCLCA3 in asthmatic mouse lung,an N-terminal 269 amino acid peptide of mCLCA3 was expressed in E.coli,purified to homogeneity and rabbit polyclonal antibodies against this peptide were generated.Immunohistochemistry of asthmatic mouse lung using the antibody indicated exclusive mCLCA3 expression in mucin granules of goblet cells in airway surface and lumen.Immunoblot analysis of lavage fluid from asthmatic mouse lung revealed a single 90 kDa protein form of mClCA3.The results demonstrate that the 90 kDa N-terminal peptide,neither the full-length protein nor the reported N-terminal 35 kDa cleaved form of mClCA3 is the major functional form involved in the packaging and exocytosis of mucin granules in asthmatic goblet cells.     

Circular RNA circRHOT1 contributes to platelet-derived growth factor BB-stimulated proliferation and migration of airway smooth muscle cells by targeting Tip60/TLR4

BackgroundAsthma serves as a chronic inflammatory respiratory diseases disorder. It mainly occurs airway inflammation and remodeling. Circular RNA (circRNA) is mainly the control of cancer disease and asthma. In specific work, we were interested in the function of circRHOT1 in the advances in asthma.MethodsEffects of platelet-derived growth factor BB (PDGF-BB) on airway smooth muscle cells (ASMCs) and remolded simulation cells were assessed. The expressions of circRHOT1, TLR4, and Tip60 were detected by qRT-PCR and the increase and decrease of cell number were analyzed by cell count-8 (CCK-8), Transwell or flow cytometry. At the same time, the levels of PCNA, IGF1R protein were determined by western blot approach. The correlation of circRHOT1, TLR4, and Tip60 was analyzed by qRT-PCR, western blotting, ChIP, and RNA pulldown.ResultsCircRHOT1 was significantly elevated in the serum collected from asthmatic patients. PDGF-BB had a positive effect on ASMCs with enhanced levels of circRHOT1, TLR4, and Tip60. Depletion of circRHOT1 avoided wider impact and migration but induced cell apoptosis. CircRHOT1 contributed PDGF-BB had a direct impact on the proliferation and migration of ASMCs by regulating TLR4. Mechanically, circRHOT1 could cooperate with acetyltransferase Tip60 in ASMCs. CircRHOT1 silencing was easy to have an adverse effect on the enrichment of Tip60 on TLR4 promoter in ASMCs. The depletion of circRHOT1 or Tip60 reduced h3k27ac and RNA polymerase II on the TLR4 promoter. Consistently, the inhibition of circRHOT1 or Tip60 reduced TLR4 expression in ASMCs. The deletion of circRHOT1 could reduce the expression level of TLR4, but the overexpression of Tip60 was easy to have a positive effect on the down-regulation of ASMCs. In addition, Tip60 could inhibit ASMCs proliferation and migration caused by PDGF-BB stimulation. CircRHOT1 could use PDGF-BB to stimulate ASMCs proliferation and migration when Tip60 regulation was implemented.ConclusionsWe can think that based on the related effects of Tip60/TLR4, circular RNA circRHOT1 with platelet-derived growth factor BB can stimulate airway smooth muscle cells to proliferate or migrate, and circRHOT1 is an important target for the treatment of asthma.     

Enhanced bioaccesibility of selective PDE4 inhibitor N-(3,4-dihydro-2h-1,5-benzodioxepin-7-yl) pyridine-4-carboxamide using polyamidoamine dendrimer

The aim of this study was to improve the aqueous solubility of highly hydrophobic but selective PDE4 inhibitor N-(3,4-dihydro-2h-1,5-benzodioxepin-7-yl) pyridine-4-carboxamide by associating it with polyamidoamine dendrimer. The PAMAM dendrimer restraining ethylenediamine core synthesized by a divergent approach was utilized for encapsulation. The solubility of conjugates was evaluated on the basis of concentration and generation of the dendrimer, pH of the solution, and temperature. The phase solubility diagram confirmed an increase in aqueous solubility of drug with increase in dendrimer concentration with respect to pH in the order 9.0 > 7.0 > 4.0. Moreover, values of thermodynamic parameters such as negative value of ΔH^o, positive value of ΔS^o and ΔG^o reflects an exothermic complexation, presence of number of particles and spontaneity of the complexation. Overall, investigations validate the enhancement in solubility of drug after complexation with polyamidoamine dendrimer which further confirmed promising bioactivity of the drug-dendrimer conjugates.     

Ultrasensitive NOX Detection in Simulated Exhaled Air: Enhanced Sensing via Alumina Modification of In‐Situ Grown WO3 Nanoblocks

Seedless growth of vertically aligned nanostructures, which can induce smoother transport and minimize Ohmic contact between substrate and semiconductor, can be fabricated by in situ growth utilizing modified hydrothermal methods. Such devices can be useful in designing non‐invasive ultrasensitive hand‐held sensors for diagnostic identification of volatile organic compounds (VOCs) in exhaled air, offering pain‐free and easier detection of long‐term diseases such as asthma. In the present work, WO₃ nanoblocks, with a high surface area and porosity, have been grown directly over transparent conducting oxide to minimize Ohmic resistance, facilitating smoother electron transfer and enhanced current response. Further modification with porous alumina (γ‐Al₂O₃), by electrodeposition, resulted in the selective and ultrasensitive detection of NO_X in simulated exhaled air. Crystal phase purity of as‐fabricated pristine as well modified samples is validated with X‐ray diffraction analysis. Morphological and microstructural analyses reveal the successful deposition of porous alumina over the surface of WO₃. Improved surface area and porosity is presented by porous alumina in the modified WO₃ device, suggesting more active sites for the gas molecules to get adsorbed and diffuse through the pores. Oxygen vacancies, which are detrimental in the transport phenomenon in the presented sensors, have been studied using X‐ray photoelectron spectroscopic (XPS) analysis. Gas sensing studies have been performed by fabricating chemiresistor devices based on bare WO₃ and Al₂O₃‐modified WO₃. The higher sensitivity for NO_X gas in case of γ‐Al₂O₃‐modified WO₃ based devices, as compared to bare WO₃‐based devices, is attributed to the better surface area and charge transport kinetics. The presented device strategy offers crucial understanding in the design and development of non‐invasive, hand‐held devices for NO gas present in the human breath, with potential application in medical diagnostics.     

Synthesis and characterization of critical process related impurities of an asthma drug – Zafirlukast

Zafirlukast is an oral leukotriene receptor antagonist (LTRA) for the treatment of pulmonary disorders such as asthma. During the process development of zafirlukast, eight process related impurities were observed at a level of 0.1–0.15 area percent. Synthesis and characterization of these impurities and investigation of the root cause of their formation is described.