MUC5AC expression is increased in bronchial submucosal glands of stable COPD patients

Aims:  Mucus expectoration is a common feature of chronic obstructive pulmonary disease (COPD). MUC5AC and MUC5B, the major mucins, are released predominantly from submucosal glands in the central airways. The aim was to investigate gland size and MUC5AC and MUC5B expression in bronchial rings from...

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Published inHistopathology Vol. 55; no. 3; pp. 321 - 331
Main Authors Caramori, Gaetano, Casolari, Paolo, Di Gregorio, Carmela, Saetta, Marina, Baraldo, Simonetta, Boschetto, Piera, Ito, Kazuhiro, Fabbri, Leonardo M, Barnes, Peter J, Adcock, Ian M, Cavallesco, Giorgio, Chung, Kian F, Papi, Alberto
Format Journal Article
LanguageEnglish
Published Oxford, UK Blackwell Publishing Ltd 01.09.2009
Blackwell
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Abstract Aims:  Mucus expectoration is a common feature of chronic obstructive pulmonary disease (COPD). MUC5AC and MUC5B, the major mucins, are released predominantly from submucosal glands in the central airways. The aim was to investigate gland size and MUC5AC and MUC5B expression in bronchial rings from smokers with COPD compared with control groups. Methods and results:  Bronchial rings from 10 non‐smoking subjects, 20 smokers with normal lung function and 20 smokers with COPD were studied. Periodic acid–Schiff (PAS) and Alcian blue histochemistry and MUC5AC and MUC5B immunohistochemistry followed by quantification of the immunoreactive area was performed. The area occupied by MUC5AC+ cells in bronchial submucosal glands was increased in COPD [20% (5.5–31.7%) gland area] compared with smokers with normal lung function [9.5% (2.5–17.5%); P < 0.05] and non‐smoking subjects [2% (0.4–6.2%); P < 0.05]. The area occupied by MUC5AC+ cells in the bronchial surface epithelium was also increased in smokers (with/without COPD) [73.5% (25–92%) epithelial area] compared with non‐smoking subjects [15% (2.7–32%); P < 0.01]. Gland size, PAS, Alcian blue staining and MUC5B expression were not significantly different among groups. MUC5AC expression correlated with the degree of airflow obstruction. MUC5AC and MUC5B expression correlated with pack‐years. Conclusions:  COPD is associated with increased MUC5AC expression in submucosal glands, indicating that MUC5AC may be involved in the pathophysiology of the disease.
AbstractList Aims:  Mucus expectoration is a common feature of chronic obstructive pulmonary disease (COPD). MUC5AC and MUC5B, the major mucins, are released predominantly from submucosal glands in the central airways. The aim was to investigate gland size and MUC5AC and MUC5B expression in bronchial rings from smokers with COPD compared with control groups. Methods and results:  Bronchial rings from 10 non‐smoking subjects, 20 smokers with normal lung function and 20 smokers with COPD were studied. Periodic acid–Schiff (PAS) and Alcian blue histochemistry and MUC5AC and MUC5B immunohistochemistry followed by quantification of the immunoreactive area was performed. The area occupied by MUC5AC+ cells in bronchial submucosal glands was increased in COPD [20% (5.5–31.7%) gland area] compared with smokers with normal lung function [9.5% (2.5–17.5%); P < 0.05] and non‐smoking subjects [2% (0.4–6.2%); P < 0.05]. The area occupied by MUC5AC+ cells in the bronchial surface epithelium was also increased in smokers (with/without COPD) [73.5% (25–92%) epithelial area] compared with non‐smoking subjects [15% (2.7–32%); P < 0.01]. Gland size, PAS, Alcian blue staining and MUC5B expression were not significantly different among groups. MUC5AC expression correlated with the degree of airflow obstruction. MUC5AC and MUC5B expression correlated with pack‐years. Conclusions:  COPD is associated with increased MUC5AC expression in submucosal glands, indicating that MUC5AC may be involved in the pathophysiology of the disease.
Mucus expectoration is a common feature of chronic obstructive pulmonary disease (COPD). MUC5AC and MUC5B, the major mucins, are released predominantly from submucosal glands in the central airways. The aim was to investigate gland size and MUC5AC and MUC5B expression in bronchial rings from smokers with COPD compared with control groups.AIMSMucus expectoration is a common feature of chronic obstructive pulmonary disease (COPD). MUC5AC and MUC5B, the major mucins, are released predominantly from submucosal glands in the central airways. The aim was to investigate gland size and MUC5AC and MUC5B expression in bronchial rings from smokers with COPD compared with control groups.Bronchial rings from 10 non-smoking subjects, 20 smokers with normal lung function and 20 smokers with COPD were studied. Periodic acid-Schiff (PAS) and Alcian blue histochemistry and MUC5AC and MUC5B immunohistochemistry followed by quantification of the immunoreactive area was performed. The area occupied by MUC5AC+ cells in bronchial submucosal glands was increased in COPD [20% (5.5-31.7%) gland area] compared with smokers with normal lung function [9.5% (2.5-17.5%); P < 0.05] and non-smoking subjects [2% (0.4-6.2%); P < 0.05]. The area occupied by MUC5AC+ cells in the bronchial surface epithelium was also increased in smokers (with/without COPD) [73.5% (25-92%) epithelial area] compared with non-smoking subjects [15% (2.7-32%); P < 0.01]. Gland size, PAS, Alcian blue staining and MUC5B expression were not significantly different among groups. MUC5AC expression correlated with the degree of airflow obstruction. MUC5AC and MUC5B expression correlated with pack-years.METHODS AND RESULTSBronchial rings from 10 non-smoking subjects, 20 smokers with normal lung function and 20 smokers with COPD were studied. Periodic acid-Schiff (PAS) and Alcian blue histochemistry and MUC5AC and MUC5B immunohistochemistry followed by quantification of the immunoreactive area was performed. The area occupied by MUC5AC+ cells in bronchial submucosal glands was increased in COPD [20% (5.5-31.7%) gland area] compared with smokers with normal lung function [9.5% (2.5-17.5%); P < 0.05] and non-smoking subjects [2% (0.4-6.2%); P < 0.05]. The area occupied by MUC5AC+ cells in the bronchial surface epithelium was also increased in smokers (with/without COPD) [73.5% (25-92%) epithelial area] compared with non-smoking subjects [15% (2.7-32%); P < 0.01]. Gland size, PAS, Alcian blue staining and MUC5B expression were not significantly different among groups. MUC5AC expression correlated with the degree of airflow obstruction. MUC5AC and MUC5B expression correlated with pack-years.COPD is associated with increased MUC5AC expression in submucosal glands, indicating that MUC5AC may be involved in the pathophysiology of the disease.CONCLUSIONSCOPD is associated with increased MUC5AC expression in submucosal glands, indicating that MUC5AC may be involved in the pathophysiology of the disease.
Mucus expectoration is a common feature of chronic obstructive pulmonary disease (COPD). MUC5AC and MUC5B, the major mucins, are released predominantly from submucosal glands in the central airways. The aim was to investigate gland size and MUC5AC and MUC5B expression in bronchial rings from smokers with COPD compared with control groups. Bronchial rings from 10 non-smoking subjects, 20 smokers with normal lung function and 20 smokers with COPD were studied. Periodic acid-Schiff (PAS) and Alcian blue histochemistry and MUC5AC and MUC5B immunohistochemistry followed by quantification of the immunoreactive area was performed. The area occupied by MUC5AC+ cells in bronchial submucosal glands was increased in COPD [20% (5.5-31.7%) gland area] compared with smokers with normal lung function [9.5% (2.5-17.5%); P < 0.05] and non-smoking subjects [2% (0.4-6.2%); P < 0.05]. The area occupied by MUC5AC+ cells in the bronchial surface epithelium was also increased in smokers (with/without COPD) [73.5% (25-92%) epithelial area] compared with non-smoking subjects [15% (2.7-32%); P < 0.01]. Gland size, PAS, Alcian blue staining and MUC5B expression were not significantly different among groups. MUC5AC expression correlated with the degree of airflow obstruction. MUC5AC and MUC5B expression correlated with pack-years. COPD is associated with increased MUC5AC expression in submucosal glands, indicating that MUC5AC may be involved in the pathophysiology of the disease.
Aims:  Mucus expectoration is a common feature of chronic obstructive pulmonary disease (COPD). MUC5AC and MUC5B, the major mucins, are released predominantly from submucosal glands in the central airways. The aim was to investigate gland size and MUC5AC and MUC5B expression in bronchial rings from smokers with COPD compared with control groups. Methods and results:  Bronchial rings from 10 non‐smoking subjects, 20 smokers with normal lung function and 20 smokers with COPD were studied. Periodic acid–Schiff (PAS) and Alcian blue histochemistry and MUC5AC and MUC5B immunohistochemistry followed by quantification of the immunoreactive area was performed. The area occupied by MUC5AC+ cells in bronchial submucosal glands was increased in COPD [20% (5.5–31.7%) gland area] compared with smokers with normal lung function [9.5% (2.5–17.5%); P  < 0.05] and non‐smoking subjects [2% (0.4–6.2%); P  < 0.05]. The area occupied by MUC5AC+ cells in the bronchial surface epithelium was also increased in smokers (with/without COPD) [73.5% (25–92%) epithelial area] compared with non‐smoking subjects [15% (2.7–32%); P  < 0.01]. Gland size, PAS, Alcian blue staining and MUC5B expression were not significantly different among groups. MUC5AC expression correlated with the degree of airflow obstruction. MUC5AC and MUC5B expression correlated with pack‐years. Conclusions:  COPD is associated with increased MUC5AC expression in submucosal glands, indicating that MUC5AC may be involved in the pathophysiology of the disease.
Author Di Gregorio, Carmela
Fabbri, Leonardo M
Cavallesco, Giorgio
Papi, Alberto
Caramori, Gaetano
Boschetto, Piera
Chung, Kian F
Barnes, Peter J
Casolari, Paolo
Saetta, Marina
Baraldo, Simonetta
Adcock, Ian M
Ito, Kazuhiro
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  givenname: Gaetano
  surname: Caramori
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  organization: Centro di Ricerca su Asma e BPCO, University of Ferrara, Ferrara, Italy
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  surname: Casolari
  fullname: Casolari, Paolo
  organization: Centro di Ricerca su Asma e BPCO, University of Ferrara, Ferrara, Italy
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  givenname: Carmela
  surname: Di Gregorio
  fullname: Di Gregorio, Carmela
  organization: Dipartimento di Anatomia-Patologica, Ospedale di Carpi (MO), Italy
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  givenname: Marina
  surname: Saetta
  fullname: Saetta, Marina
  organization: Department of Cardio-Thoracic and Vascular Sciences, Section of Respiratory Diseases, University of Padova, Padova, Italy
– sequence: 5
  givenname: Simonetta
  surname: Baraldo
  fullname: Baraldo, Simonetta
  organization: Department of Cardio-Thoracic and Vascular Sciences, Section of Respiratory Diseases, University of Padova, Padova, Italy
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  givenname: Piera
  surname: Boschetto
  fullname: Boschetto, Piera
  organization: Igiene e Medicina del Lavoro e, University of Ferrara, Ferrara, Italy
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  givenname: Kazuhiro
  surname: Ito
  fullname: Ito, Kazuhiro
  organization: Airways Disease Section, NHLI, Imperial College London, London, UK
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  givenname: Leonardo M
  surname: Fabbri
  fullname: Fabbri, Leonardo M
  organization: Section of Respiratory Disease, Department of Oncology, Haematology and Respiratory Disease, University of Modena and Reggio Emilia, Modena
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  givenname: Peter J
  surname: Barnes
  fullname: Barnes, Peter J
  organization: Airways Disease Section, NHLI, Imperial College London, London, UK
– sequence: 10
  givenname: Ian M
  surname: Adcock
  fullname: Adcock, Ian M
  organization: Airways Disease Section, NHLI, Imperial College London, London, UK
– sequence: 11
  givenname: Giorgio
  surname: Cavallesco
  fullname: Cavallesco, Giorgio
  organization: Chirurgia Toracica, University of Ferrara, Ferrara, Italy
– sequence: 12
  givenname: Kian F
  surname: Chung
  fullname: Chung, Kian F
  organization: Airways Disease Section, NHLI, Imperial College London, London, UK
– sequence: 13
  givenname: Alberto
  surname: Papi
  fullname: Papi, Alberto
  organization: Centro di Ricerca su Asma e BPCO, University of Ferrara, Ferrara, Italy
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IsDoiOpenAccess false
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Issue 3
Keywords Human
Lung disease
Respiratory disease
Mucin 5AC
MUCSAC
Mucus
Bronchial gland
MUC5B
submucosal bronchial glands
Anatomic pathology
Chronic
Bronchus disease
Submucosa
Bronchitis
Chronic obstructive pulmonary disease
chronic bronchitis
Obstructive pulmonary disease
COPD
Language English
License CC BY 4.0
LinkModel DirectLink
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OpenAccessLink http://hdl.handle.net/11380/618035
PMID 19723147
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PublicationDate September 2009
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  text: September 2009
PublicationDecade 2000
PublicationPlace Oxford, UK
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PublicationTitle Histopathology
PublicationTitleAlternate Histopathology
PublicationYear 2009
Publisher Blackwell Publishing Ltd
Blackwell
Publisher_xml – name: Blackwell Publishing Ltd
– name: Blackwell
References Innes AL, Woodruff PG, Ferrando RE et al. Epithelial mucin stores are increased in the large airways of smokers with airflow obstruction. Chest 2006; 130; 1102-1108.
Chen Y, Thai P, Zhao YH, Ho YS, DeSouza MM, Wu R. Stimulation of airway mucin gene expression by interleukin (IL)-17 through IL-6 paracrine/autocrine loop. J. Biol. Chem. 2003; 278; 17036-17043.
Caramori G, Casolari P, Saetta M et al. MUC5AC and MUC5B expression in central airways from non-smokers, smokers with normal lung function and GOLD stage 1 and 2 COPD patients. Eur. Respir. J. 2006; 28(Suppl. 50); 663s abstract P3849.
Hovenberg HW, Davies JR, Carlstedt I. Different mucins are produced by the surface epithelium and the submucosa in human trachea: identification of MUC5AC as a major mucin from the goblet cells. Biochem. J. 1996; 318; 319-324.
Baraldo S, Bazzan E, Turato G et al. Decreased expression of TGF-beta type II receptor in bronchial glands of smokers with COPD. Thorax 2005; 60; 998-1002.
Burgel PR, Montani D, Danel C, Dusser DJ, Nadel JA. A morphometric study of mucins and small airway plugging in cystic fibrosis. Thorax 2007; 62; 153-161.
Sommerhoff CP, Finkbeiner WE. Human tracheobronchial submucosal gland cells in culture. Am. J. Respir. Cell Mol. Biol. 1990; 2; 41-50.
Global Initiative for Chronic Obstructive Lung Disease. Global strategy for the diagnosis, management and prevention of chronic obstructive pulmonary disease. NHLBI/WHO workshop report. NIH Publication No 2701. Bethesda, MD: National Heart, Lung and Blood Institute, 2001 (last update 2008); 1-100. Available at http://www.goldcopd.com (last accessed 11 August 2009).
Caramori G, Di Gregorio C, Carlstedt I et al. Mucin expression in peripheral airways of patients with chronic obstructive pulmonary disease. Histopathology 2004; 45; 477-484.
Rose MC, Voynow JA. Respiratory tract mucin genes and mucin glycoproteins in health and disease. Physiol. Rev. 2006; 86; 245-278.
Rogers DF, Barnes PJ. Treatment of airway mucus hypersecretion. Ann. Med. 2006; 38; 116-125.
Vestbo J, Hogg JC. Convergence of the epidemiology and pathology of COPD. Thorax 2006; 61; 86-88.
Saetta M, Turato G, Maestrelli P, Mapp CE, Fabbri LM. Cellular and structural bases of chronic obstructive pulmonary disease. Am. J. Respir. Crit. Care Med. 2001; 163; 1304-1309.
Di Stefano A, Caramori G, Oates T et al. Increased expression of NF-κB in bronchial biopsies from smokers and patients with chronic obstructive pulmonary disease (COPD). Eur. Respir. J. 2002; 20; 556-563.
Sethi S, Maloney J, Grove L, Wrona C, Berenson CS. Airway inflammation and bronchial bacterial colonization in chronic obstructive pulmonary disease. Am. J. Respir. Crit. Care Med. 2006; 173; 991-998.
Papi A, Bellettato CM, Braccioni F et al. Infections and airway inflammation in chronic obstructive pulmonary disease severe exacerbations. Am. J. Respir. Crit. Care Med. 2006; 173; 1114-1121.
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Ten Hacken NH, Aleva RM, Rutgers B, Kraan J, Van Goor H, Postma DS. Techniques in human airway inflammation: differences in plastic-embedded and snap-frozen sections for CD3, CD4, and CD8 immunostaining of bronchial biopsy specimens. Mod. Pathol. 1997; 10; 1043-1046.
Hogg JC, Chu F, Utokaparch S et al. The nature of small-airway obstruction in chronic obstructive pulmonary disease. N. Engl. J. Med. 2004; 350; 2645-2653.
Copin MC, Buisine MP, Devisme L et al. Normal respiratory mucosa, precursor lesions and lung carcinomas: differential expression of human mucin genes. Front. Biosci. 2001; 6; D1264-D1275.
Reid LM. Measurement of the bronchial mucous gland layer: a diagnostic yardstick in chronic bronchitis. Thorax 1960; 15; 132-141.
De Marco R, Accordini S, Cerveri I et al. Incidence of COPD in a cohort of young adults according to the presence of chronic cough and phlegm. Am. J. Respir. Crit. Care Med. 2007; 175; 32-39.
Thai P, Loukoianov A, Wachi S, Wu R. Regulation of airway mucin gene expression. Annu. Rev. Physiol. 2008; 70; 405-429.
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References_xml – reference: O'Donnell RA, Richter A, Ward J et al. Expression of ErbB receptors and mucins in the airways of long term current smokers. Thorax 2004; 59; 1032-1040.
– reference: Schulz BL, Sloane AJ, Robinson LJ et al. Mucin glycosylation changes in cystic fibrosis lung disease are not manifest in submucosal gland secretions. Biochem. J. 2005; 387; 911-919.
– reference: Global Initiative for Chronic Obstructive Lung Disease. Global strategy for the diagnosis, management and prevention of chronic obstructive pulmonary disease. NHLBI/WHO workshop report. NIH Publication No 2701. Bethesda, MD: National Heart, Lung and Blood Institute, 2001 (last update 2008); 1-100. Available at http://www.goldcopd.com (last accessed 11 August 2009).
– reference: Varani K, Caramori G, Vincenzi F et al. Alteration of adenosine receptors in patients with chronic obstructive pulmonary disease. Am. J. Respir. Crit. Care Med. 2006; 173; 398-406.
– reference: Hogg JC, Chu F, Utokaparch S et al. The nature of small-airway obstruction in chronic obstructive pulmonary disease. N. Engl. J. Med. 2004; 350; 2645-2653.
– reference: Caramori G, Di Gregorio C, Carlstedt I et al. Mucin expression in peripheral airways of patients with chronic obstructive pulmonary disease. Histopathology 2004; 45; 477-484.
– reference: Chen Y, Thai P, Zhao YH, Ho YS, DeSouza MM, Wu R. Stimulation of airway mucin gene expression by interleukin (IL)-17 through IL-6 paracrine/autocrine loop. J. Biol. Chem. 2003; 278; 17036-17043.
– reference: Sethi S, Maloney J, Grove L, Wrona C, Berenson CS. Airway inflammation and bronchial bacterial colonization in chronic obstructive pulmonary disease. Am. J. Respir. Crit. Care Med. 2006; 173; 991-998.
– reference: Vestbo J, Hogg JC. Convergence of the epidemiology and pathology of COPD. Thorax 2006; 61; 86-88.
– reference: Caramori G, Casolari P, Saetta M et al. MUC5AC and MUC5B expression in central airways from non-smokers, smokers with normal lung function and GOLD stage 1 and 2 COPD patients. Proc. Am. Thorac. Soc. 2006; 3 (abstracts issue); abstract A618.
– reference: Papi A, Bellettato CM, Braccioni F et al. Infections and airway inflammation in chronic obstructive pulmonary disease severe exacerbations. Am. J. Respir. Crit. Care Med. 2006; 173; 1114-1121.
– reference: Copin MC, Buisine MP, Devisme L et al. Normal respiratory mucosa, precursor lesions and lung carcinomas: differential expression of human mucin genes. Front. Biosci. 2001; 6; D1264-D1275.
– reference: Thai P, Loukoianov A, Wachi S, Wu R. Regulation of airway mucin gene expression. Annu. Rev. Physiol. 2008; 70; 405-429.
– reference: Caramori G, Casolari P, Saetta M et al. MUC5AC and MUC5B expression in central airways from non-smokers, smokers with normal lung function and GOLD stage 1 and 2 COPD patients. Eur. Respir. J. 2006; 28(Suppl. 50); 663s abstract P3849.
– reference: Hovenberg HW, Davies JR, Carlstedt I. Different mucins are produced by the surface epithelium and the submucosa in human trachea: identification of MUC5AC as a major mucin from the goblet cells. Biochem. J. 1996; 318; 319-324.
– reference: Alexis NE, Hu SC, Zeman K, Alter T, Bennett WD. Induced sputum derives from the central airways: confirmation using a radiolabeled aerosol bolus delivery technique. Am. J. Respir. Crit. Care Med. 2001; 164; 1964-1970.
– reference: Saetta M, Turato G, Maestrelli P, Mapp CE, Fabbri LM. Cellular and structural bases of chronic obstructive pulmonary disease. Am. J. Respir. Crit. Care Med. 2001; 163; 1304-1309.
– reference: Di Stefano A, Caramori G, Oates T et al. Increased expression of NF-κB in bronchial biopsies from smokers and patients with chronic obstructive pulmonary disease (COPD). Eur. Respir. J. 2002; 20; 556-563.
– reference: Quanjer P, Tammeling GJ, Cotes JE, Pedersen OF, Peslin R, Yernault JC. Lung volumes and forced ventilatory flows. Report Working Party, standardization of lung function tests, European Community for Steel and Coal, official statement of the European Respiratory Society. Eur. Respir. J. 1993; 6(Suppl. 16); 5-40.
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– reference: Innes AL, Woodruff PG, Ferrando RE et al. Epithelial mucin stores are increased in the large airways of smokers with airflow obstruction. Chest 2006; 130; 1102-1108.
– reference: Sommerhoff CP, Finkbeiner WE. Human tracheobronchial submucosal gland cells in culture. Am. J. Respir. Cell Mol. Biol. 1990; 2; 41-50.
– reference: Rogers DF, Barnes PJ. Treatment of airway mucus hypersecretion. Ann. Med. 2006; 38; 116-125.
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– reference: Burgel PR, Montani D, Danel C, Dusser DJ, Nadel JA. A morphometric study of mucins and small airway plugging in cystic fibrosis. Thorax 2007; 62; 153-161.
– reference: Rose MC, Voynow JA. Respiratory tract mucin genes and mucin glycoproteins in health and disease. Physiol. Rev. 2006; 86; 245-278.
– reference: Baraldo S, Bazzan E, Turato G et al. Decreased expression of TGF-beta type II receptor in bronchial glands of smokers with COPD. Thorax 2005; 60; 998-1002.
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Snippet Aims:  Mucus expectoration is a common feature of chronic obstructive pulmonary disease (COPD). MUC5AC and MUC5B, the major mucins, are released predominantly...
Aims:  Mucus expectoration is a common feature of chronic obstructive pulmonary disease (COPD). MUC5AC and MUC5B, the major mucins, are released predominantly...
Mucus expectoration is a common feature of chronic obstructive pulmonary disease (COPD). MUC5AC and MUC5B, the major mucins, are released predominantly from...
SourceID proquest
pubmed
pascalfrancis
crossref
wiley
istex
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Index Database
Enrichment Source
Publisher
StartPage 321
SubjectTerms Aged
Alcian Blue
Biological and medical sciences
Bronchi - metabolism
Bronchi - pathology
chronic bronchitis
Chronic obstructive pulmonary disease, asthma
COPD
Female
Humans
Investigative techniques, diagnostic techniques (general aspects)
Male
Medical sciences
MUC5AC
MUC5B
Mucin 5AC - metabolism
Mucin-5B - metabolism
Mucins - metabolism
mucus
Pathology. Cytology. Biochemistry. Spectrometry. Miscellaneous investigative techniques
Pneumology
Pulmonary Disease, Chronic Obstructive - etiology
Pulmonary Disease, Chronic Obstructive - metabolism
Pulmonary Disease, Chronic Obstructive - pathology
Pulmonary Disease, Chronic Obstructive - physiopathology
Respiratory Function Tests
Respiratory Mucosa - metabolism
Smoking - adverse effects
Staining and Labeling
submucosal bronchial glands
Title MUC5AC expression is increased in bronchial submucosal glands of stable COPD patients
URI https://api.istex.fr/ark:/67375/WNG-WT2M7PRH-M/fulltext.pdf
https://onlinelibrary.wiley.com/doi/abs/10.1111%2Fj.1365-2559.2009.03377.x
https://www.ncbi.nlm.nih.gov/pubmed/19723147
https://www.proquest.com/docview/67632237
Volume 55
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