otolaryngologia polska 67 (2013) 192–197
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Original research article/Artykuł oryginalny
Lysosomal exoglycosidases in nasal polyps Sylwia Chojnowska 1,*, Alina Minarowska 2, Małgorzata Knaś 3, Anna Niemcunowicz-Janica 4, Paweł Kołodziejczyk 5, Beata Zalewska-Szajda 6, Alina Kępka 7, Łukasz Minarowski 8, Napoleon Waszkiewicz 9, Krzysztof Zwierz 5, Sławomir Dariusz Szajda 9 1
Medical Institute, College of Computer Science and Business Administration, Lomza, Poland Department of Anatomy, Poland 3 Research Laboratory of Cosmetology, Poland 4 Department of Forensic Medicine, Poland 5 Medical College of the Universal Education Society, Lomza, Poland 6 Department of Radiology, Medical University Children Hospital, Bialystok, Poland 7 The Children's Memorial Health Institute, Warsaw, Poland 8 Department of Lung Diseases and Tuberculosis, Medical University of Bialystok, Poland 9 Department of the Emergency Medicine and Disasters, Medical University of Bialystok, Poland 2
article info
abstract
Article history:
Introduction: Nasal polyps are smooth outgrowths assuming a shape of grapes, formed
Received: 14.05.2013
from the nasal mucosa, limiting air flow by projecting into a lumen of a nasal cavity. Up
Accepted: 22.05.2013
to now the surgical resection is the best method of their treatment, but etiology and
Available online: 24.05.2013
pathogenesis of the nasal polyps is not yet fully established. Aim of the study: The aim of the study was the assessment of the selected lysosomal exoglycosidases activity in
Keywords: Lysosomal exoglycosidases a-Fucosidase
the nasal polyps. In this study the activity of b-galactosidase, a-mannosidase and a-
b-Galactosidase a-Mannosidase
during mucotomy from 20 patients (10 F, 10 M). Results: We observed significant lower
Nasal polyps
polyps (P) in comparison to control healthy nasal mucosa (C). In nasal polyp tissue (P) no
fucosidase was determined in the tissue of the nasal polyps obtained from 40 patients (10 F, 30 M) and control tissues derived from mucosa of lower nasal conchas obtained values of GAL, FUC and tendency to decrease of MAN and GLU concentration in nasal differences of GAL, MAN and FUC specific activity in comparison to control mucosa (C) were found. Conclusions: Our research supports bioelectrical theory of the nasal polyps pathogenesis and directs attention at research on glycoconjugates and glycosidases of the nasal mucosa extracellular matrix. © 2013 Polish Otorhinolaryngology - Head and Neck Surgery Society. Published by Elsevier Urban & Partner Sp. z o.o. All rights reserved.
* Corresponding author at: Medical Institute, College of Computer Science and Business Administration, Akademicka Str. 14, 18-400 Łomża, Poland. Tel.: +48 86 215 59 53; fax: +48 86 215 66 01. E-mail address:
[email protected] (S. Chojnowska). 0030-6657/$ – see front matter © 2013 Polish Otorhinolaryngology - Head and Neck Surgery Society. Published by Elsevier Urban & Partner Sp. z o.o. All rights reserved.
http://dx.doi.org/10.1016/j.otpol.2013.05.004
otolaryngologia polska 67 (2013) 192–197
Introduction Nasal polyps (gr. polýpous – polypod, i.e. many pads) [1] are smooth, grapes shape outgrowths of the nasal mucosa, projecting into nasal cavity, reducing nasal air flow, and creating danger of extra- and intra-cranial complications [2– 4]. Nasal polyps affect from 0.2–5% to 28% of the world population, mostly men in Europe, after 20 years of life with a mean age 38–39 years [5–9]. Nasal polyps accompanying by eosinophilia consist 80–90% of all nasal polyps, and the rest is accompanied by neutrophilia [10–12]. Local application of glucocorticosteroids is effective in relieving symptoms of the eosinophilic nasal polyps, decreasing their size and usually preventing against relapse, but neutrophilic polyps did not answer corticosteroid treatment [13, 14]. Recently new therapies are being tested for nasal polyps treatment: antiallergic drugs with monoclonal antibodies against IgE and antibiotics directed against bacterial biofilm [4, 15–19]. In the nasal polyps treatment, the best results obtained by polypectomy, particularly in large polyps and polyps occupying nasal sinuses [20, 21]. However in up to 87% of polypectomised patients recurrence of was observed [3, 18]. Majority of physicians believe that nasal polyps are final stage of a chronic inflammatory process in the nasal mucosal membrane [22–24]. In 1994 Bernstein proposed inflammatorybioelectric theory of nasal polyps formation based on abnormal chloride channels favoring increased absorption of sodium cations, changed composition of nasal mucus, increased water penetration to nasal mucosa intracellular matrix, swelling of mucosal tissue [9, 27]. The Bernstein theory is in agreement with reported higher (in comparison to healthy nasal mucosa) ability of the nasal polyps tissue to absorb Na+ [27, 28]. Recently attempts to correlate nasal polyps incidence with eosinophilia and eosinophilic inflammation of mucosa were reported [10, 11]. However, it was reported that only 5% of persons with nasal polyps suffer from allergy, with exception of fungal allergy (allergic fungal rhinosinusitis – AFRS) where frequency of nasal polyps exceeded 85% [2, 7, 25, 26]. It is generally accepted that in rhinosinuitis with nasal polyps formation following mechanisms are involved: excessive fibroblast proliferation after injury, degranulation of mastocytes, histamine release, eosinophiles infiltration and edema with exudation [7, 28]. It appears that, independently of etiology nasal polyps formation should be related to changes in structure, concentration and metabolism of substances building nasal mucosa and among them glycoconjugates (glycoproteins, glycolipids, and proteoglycans) constituting cell membranes, extracellular matrix and biofilm covering nasal mucosa [17, 19]. In catabolism of glycoconjugates saccharide chains lysosomal exoglycosidases are involved [37]. Thus, the aim of our work was an evaluation of selected lysosomal exoglycosidases in pathogenesis of the nasal polyps.
Materials and methods Patients. Nasal polyps were obtained during polypectomy from 40 patients (10 women and 30 men, aged 23–84 years)
193
with diagnosed chronic rhinosinusitis with nasal polyps. Control tissues were derived from mucosa of lower nasal conchas obtained during mucotomy operations from 20 patients (10 women and 10 men, aged 23–61 years). Biological material processing. To perform histopathological examination, fragments of polyps and normal nasal tissue were immersed in 10% formalin and subsequently stained with H + E. For biochemical investigations fragments of polyps and normal nasal mucosa were homogenized in homogenizer (Ultra-Turrax T8, Germany). Homogenates were centrifuged at 12 000 g for 20 min at 4 8C before further procedures. Glycosidases activity. Activity of exoglycosidases in supernatants of centrifuged homogenates was determined by Chatterjee et al. method [29] in modification of Zwierz et al. [30] and Marciniak et al. [31] with 4-nitrophenylderivatives of the appropriate sugars. Total protein concentration was determined by the Lowry et al. method [32]. Ethical issues. Informed written consent was obtained from all the participants after explanation of the nature, purpose, and potential risk of the study. The study was approved by the Bioethical Committee of the Medical University of Białystok, Poland (R-I-002/46/2007). Statistical analysis. All variables were checked for normality. Results were expressed as the mean SD. P values less than 0.05 were considered significant. Statistical analysis was performed using packet Statistica 6.0 (StatSoft, Poland).
Results Normal nasal mucosa and nasal polyp tissue with pathological findings (submucosal edema, inflammatory infiltration, superficial hyperplasia) are presented in Fig. 1. We observed significant lower values of GAL, FUC and tendency to decrease MAN and GLU concentration in nasal polyps (P) in comparison to controls (C) (Fig. 2). In nasal polyp tissue (P) no differences of GAL, MAN and FUC specific activity in comparison to control mucosa were observed (C) (Fig. 3). It is worthy to note a similar decrease in concentration of activity for GAL, MAN, and FUC (from 0.71 for FUC to 0.78 for MAN) in polypous tissue in comparison to controls (Table I) and the same level of tendency to decrease in specific activity, i.e. from 0.79 to FUC to 0.92 for MAN (Table I).
Discussion In pathological situations connected with tissue remodeling such as colorectal cancer, pancreatic cancer, renal cancer, salivary gland tumors [36], alcoholism [37], Lyme disease [50] significant increase in lysosomal exoglycosidases activity was observed. Decrease in concentration of the activity of lysosomal exoglycosidases in nasal polyps tissue is contraindicatory to active inflammatory process in nasal polyps. In nasal polyps we observed significant decrease in concentration the activity of GAL, FUC, and tendency to
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otolaryngologia polska 67 (2013) 192–197
[(Fig._1)TD$IG]
Fig. 1 – (A) Normal mucosa (H + E, magnification 20T). (B). Submucosal edema of polypous tissue (H + E, magnification 10T). (C) Polypous tissue with inflammation foci (H + E, magnification 10T). (D) Stratified squamous epithelium hyperplasia on the surface of a polyp (H + E, magnification 20T)
decrease MAN concentration (Table I, Fig. 2). In polypous tissue, in comparison to control nasal mucosa, specific activity of GAL, MAN, and FUC had tendency to decrease (Table II, Fig. 3). Decrease in concentration of GAL and FUC in association with tendency to decrease in concentration of MAN and all exoglycosidases specific activities is in agreement with accumulation of water in polypous tissue. Our
results are contradictory to existence of full symptomatic eosinophilic [11, 33] or neutrophilic [21, 33] inflammation in nasal polyps, as inflammatory state is connected with significant increase in activity of lysosomal exoglycosidases [37]. It is worthy to note that level of decrease in exoglycosidases concentration at nasal polyps tissue was similar for all exoglycosidases tested, e.g. from 0.71 for FUC to 0.78 for
[(Fig._2)TD$IG]
[(Fig._3)TD$IG]
Fig. 2 – Lysosomal exoglycosidases concentrations in nasal polyps and control tissues; c – control, p – polyp, *p < 0.05, **p < 0.01
Fig. 3 – Lysosomal exoglycosidases specific activities in nasal polyps and control tissues; c – control, p – polyp
otolaryngologia polska 67 (2013) 192–197
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Table I – Proportions of lysosomal exoglycosidases concentrations in nasal polyps and control tissues Exoglycosidase
Tissue
N
Concentration of activity (pKat/1 g tissue)
p
GAL
control polyp control polyp control polyp
20 40 20 40 20 40
259.1 93.3 195.7 89.9 194.7 71.5 138.6 72.2 185.2 84.3 143.9 74.4
0.017
FUC MAN
MAN (Table I), which suggest similar dilution of the exoglycosidases activity in the nasal polyps tissue. Of exoglycosidases tested in nasal polyps only concentration activity of MAN (taking part in degradation of N-linked oligosaccharides) did not significantly decrease, which suggest weaker involvement of N-linked glycoproteins in nasal polyps pathology. Full inflammatory response includes production of the inflammation mediators such as prostaglandins [34] and cytokines [35] responsible for local inflammatory reaction. Decrease in concentration of exoglycosidases and presence of some inflammatory cells in nasal polyps, i.e. acidophilic and neutrophilic granulocytes suggest deregulation of the mechanisms of inflammation. In full symptomatic inflammatory processes, e.g. tonsillitis [38] was reported increase in tissue remodeling expressed by increase in lysosomal catabolism of glycoconjugates, reflected by increase in concentration of activity the lysosomal exoglycosidases. Decrease in concentration of exoglycosidases activity participating in the catabolism of oligosaccharide chains of glycoconjugates in nasal polyps tissue (Figs. 2 and 3) is an argument against recognition nasal polyps as a fully inflammatory disease. Our results confirm inflammatory-bioelectrical theory of nasal polyp pathogenesis [17, 27, 28]. Our results corroborate the decrease in secretion of cytokines, expression of adhesion molecules, amount of eosinophiles and mastocytes observed in polyps tissue [39–43]. Glucocorticosteroid application in prevention of nasal polyps recurrence suggests involvement of chronic inflammatory immunological mechanism deregulation in pathogenesis of the nasal polys [44, 45]. Our results agree with results of Metzler et al. [46], that inflammation precedes nasal polyps, by accumulation of fluids and albumins in subepithelial layer. RostkowskaNadolska et al. [45, 47] reported that cytokeratines 4,13, and 19 expressions suggest involvement in nasal polyps rather stroma than epithelium. Our results as well as lack of cells edema in nasal polyps tissue [18], and excessive hydratation of nasal polyps extracellular matrix are in agreement with
0.007 0.072
Bernstein and Yankaskas suggestion [48, 49], that one of the factors responsible for nasal polyps creation are disturbances in transport of water and electrolytes in nasal mucosa. Bernstein and Yankaskas proposed the abnormal Cystic Fibrosis Transmembrane Regulator (CFTR) protein, regulating sodium channels (ENaC), for increase in amount of open sodium channels on the surface of nasal mucosa endothelial cells and stromal swelling [48]. For absorption of sodium cations in stroma also Major Basic Proteins may be responsible, secreted by eosinophils, which decrease secretion of mucus and increase absorption of sodium [17, 22]. Higher ability to absorb sodium and chloride ions by nasal polyps mucosa supports the opinion of significant role of deregulation water and mineral metabolism in pathogenesis of nasal polyps [27]. In extracellular matrix main substances responsible for water absorption are hyaluronate and proteoglycans (e.g. chondroitin sulfate), because of their negative charged polyanion and hydrophilic –OH groups [50]. It is worthy of note that only small part of water molecules are directly bound to the proteoglycans by hydrogen bounds, but majority of water molecules is mechanically trapped inside of heteropolysaccharide structure of hyaluronate [51, 52]. Therefore it would be a valuable information on composition and distribution of glycosaminoglycans and proteoglycans in nasal polyps tissue in comparison to normal nasal mucosa. In conclusion, our research supports bioelectrical theory of nasal polyps pathogenesis and directs attention at research on glycoconjugates (particularly glycosaminoglycans) of the nasal mucosa extracellular matrix, the structural elements responsible for absorption the excessive amounts of water.
Authors' contributions/Wkład autorów SC – study design, data collection and interpretation, acceptance of final manuscript version, literature search. AM – study design, data collection and interpretation, acceptance
Table II – Proportions of lysosomal exoglycosidases specific activities in nasal polyps and control tissues Exoglycosidase
Tissue
N
Specific activity (pKat/mg protein)
p
GAL
control polyp control polyp control polyp
20 40 20 40 20 40
83.3 24.9 73.6 45.6 65.1 27.8 51.5 31.0 58.9 22.2 54.3 36.2
0.292
FUC MAN
0.093 0.544
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of final manuscript version. MK, AN-J – data collection and interpretation, acceptance of final manuscript version. PK – statistical analysis, data interpretation, acceptance of final manuscript version. BZ-S – data collection, acceptance of final manuscript version. AK, NW – data interpretation, acceptance of final manuscript version. ŁM – statistical analysis, data interpretation, literature search. KZ – study design, data interpretation, acceptance of final manuscript version, funds collection. SDS – study design, data interpretation, acceptance of final manuscript version.
Conflict of interest/Konflikt interesu None declared.
Financial support/Finansowanie The study was financed from the grant of Medical University of Bialystok No. R-I-002/46/2007.
Ethics/Etyka The work described in this article have been carried out in accordance with The Code of Ethics of the World Medical Association (Declaration of Helsinki) for experiments involving humans; EU Directive 2010/63/EU for animal experiments; Uniform Requirements for manuscripts submitted to Biomedical journals.
r e f e r e n c e s / p i s m i e n n i c t w o
[1] Mullol J. Trends on rhinosinusitis diagnosis and treatment. Otolaryngol Pol 2009;63:3–4. [2] Zakrzewska A, Piotrowska V, Zieliński R. Pathologies of lateral nasal wall in children with diagnosed antrochoanal polyps. Otolaryngol Pol 2011;65:208–213. [3] Muñoz-Del-Castillo F, Jurado-Ramos A, Soler R, FernándezConde BL, Barasona MJ, Cantillo E, et al. Fungal sensitization in nasal polyposis. J Investig Allergol Clin Immunol 2009;19:6–12. [4] Naclerio RM, Mackay IS. Guidelines for the management of nasal polyposis. In: Mygind N, Linholdt T, editors. Nasal polyposis. An inflammatory disease and its treatment. Copenhagen: Munksgaard; 1997. p. 177–180. [5] Jurkiewicz D, Rapiejko P. Use of isotonic NaCl solution in patients with acute rhinosinusitis. Otolaryngol Pol 2011;65:47–53. [6] Toledano Muñoz A, Herráiz Puchol C, Navas Molinero C, García Simal M, Navarro Cunchillos M, Galindo Campillo AN. Epidemiological study in patients with nasal polyposis. Acta Otorrinolaringol Esp 2008;59:438–443. [7] Jurkiewicz D. Nasal polyps. Mag Otorynolaryngol 2003;11:3–9. [8] Rugina M, Serrano E, Klossek JM, Crampette L, Stoll D, Bebear JP, et al. Epidemiological and clinical aspects of nasal polyposis in France; the ORLI group experience. Rhinology 2002;40:75–79. [9] Bernstein JM, Gorfien J, Noble B. Role of allergy in nasal polyposis. Otolaryngol Head Neck Surg 1995;113:724–732.
[10] Armengot M, Garín L, de Lamo M, Krause F, Carda C. Cytological and tissue eosinophilia correlations in nasal polyposis. Am J Rhinol Allergy 2010;24:413–415. [11] Perić A, Vojvodić D, Vukomanović-Đurđević B, Baletić N. Eosinophilic inflammation in allergic rhinitis and nasal polyposis. Arh Hig Rada Toksikol 2011;62:341–348. [12] Miłoński J, Zielińska-Bliźniewska H, Pietkiewicz P, Olszewski J. Analysis of histopathological evaluation of pathological lesions removed by endoscopic surgery of the nose and paranasal sinuses in the own material. Otolaryngol Pol 2011;65:447–450. [13] Modrzyński M, Zawisza E. Place of steroid therapy in the treatment of nasal polyps in the light of recent years reports. Pol Merkuriusz Lek 2000;7:51–54. [14] Rostkowska-Nadolska B, Mazurek U, Kapral M. Pharmacotherapy of nasal polyp. Ann Acad Med Siles 2006;6:163–166. [15] Mrówka-Kata K, Czecior E, Kata D, Namysłowski G, Dziechciarz-Werbowska J, Sowa P. Current view on nasal polyps management in Samter's triad patients. Otolaryngol Pol 2012;66:373–378. [16] Penn R, Mikula S. The role of anti-IgE immunoglobulin therapy in nasal polyposis: a pilot study. Am J Rhinol 2007;21:428–432. [17] Bernstein JM. Update on the molecular biology of nasal polyposis. Otolaryngol Clin North Am 2005;38:1243–1255. [18] De Castro MC, Assuncao E, Moreira de Castro M, Araújo RN, Guimarães RE, Nunes FB. Effect of mitomocin C in eosinophilic nasal polyposis, in vivo: concentration of IL5 and GM-CSF, RT-PCR. Rev Bras Otorrinolaringol 2006;71: 38–42. [19] Górski NP, Palmer JN. Bacterial biofilms in chronic rhinosinusitis. Mag Otorynolaryngol 2006;6(Suppl. 9):24–30. [20] Devars du Mayne M, Prulière-Escabasse V, Zerah-Lancner F, Coste A, Papon JF. Polypectomy compared with ethmoidectomy in the treatment of nasal polyposis. Arch Otolaryngol Head Neck Surg 2011;137:111–117. [21] Gromek I, Krzeski A. Neutrophilic and eosinophilic chronic sinusitis, nasal polyps. Mag Otorynolaryngol 2006;6(Suppl. 9):11–23. [22] Jordana M, Dolovich J, Ohno I, Finotto S, Denburg J. Nasal polyposis model of chronic inflammation. In: Busse WW, Holgate ST, editors. Astma and rhinitis. Boston: Blackwell Scientific Publication; 1995. p. 156–164. [23] Sachse F, Becker K, Basel TJ, Weiss D, Rudack C. IKK-2 inhibitor TPCA-1 represses nasal epithelial inflammation in vitro. Rhinology 2011;49:168–173. [24] Zhang G, Jing X, Wang X, Shi W, Sun P, Su C, et al. Contribution of the proinflammatory cytokine IL-18 in the formation of human nasal polyps. Anat Rec (Hoboken) 2011;294:953–960. [25] Muñoz del Castillo F, Jurado-Ramos A, Fernández-Conde BL, Soler R, Barasona MJ, Cantillo E, et al. Allergenic profile of nasal polyposis. J Investig Allergol Clin Immunol 2009;19:110–116. [26] Mulligan JK, Bleier BS, O'Connell B, Mulligan RM, Wagner C, Schlosser RJ. Vitamin D3 correlates inversely with systemic dendritic cell numbers and bone erosion in chronic rhinosinusitis with nasal polyps and allergic fungal rhinosinusitis. Clin Exp Immunol 2011;164:312–320. [27] Al-Bazzaz F, Yadawa VP, Westenfelder C. Modification of Na+ and Cl transport in canine tracheal mucosa by prostaglandins. Am J Physiol 1981;240:101–105. [28] Bernstein JM. The molecular biology of nasal polyposis. Curr Allergy Asthma Rep 2001;1:262–267. [29] Chatterjee S, Velicer LF, Sweeley CC. Glycosphingolipid glycosyl hydrolases and glycosidases of synchronized human KB cells. J Biol Chem 1975;250:4972–4979.
otolaryngologia polska 67 (2013) 192–197
[30] Chlabicz M, Sieśkiewicz A, Rózańska-Kudelska M, Olszewska E, Roszkowska-Jakimiec W, Rogowski M, et al. Cathepsin D activity in chronic rhinosinusitis with nasal polyps. Otolaryngol Pol 2010;64:299–301. [31] Marciniak J, Zalewska A, Popko J, Zwierz K. Optimization of an enzymatic method for the determination of lysosomal N-acetyl-beta-hexosaminidase and beta-glucuronidase in synovial fluid. Clin Chem Lab Med 2006;44:933–937. [32] Lowry OH, Rosebrough NJ, Farr AL, Randal RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193:265–275. [33] Miłoński J, Zielińska-Bliźniewska H, Sobański R, Olszewski J. The comparison of the influence of various types of anaesthesia on perioperative bleeding control in endoscopic paranasal sinus surgery. Otolaryngol Pol 2012;66:122–125. [34] Kawano M, Okada K, Muramoto H, Morishita H, Omura T, Inoue R. Simultaneous, clonally identical T cell expansion in tonsil and synovium in a patient with rheumatoid arthritis and chronic tonsillitis. Arthritis Rheum 2003;48:2483–2488. [35] Rostkowska-Nadolska B, Pośpiech L, Preś K. The role of cytokines in nasal polyps. Otorynolaryngology 2006;5:1–6. [36] Bierc M, Minarowski L, Woźniak L, Chojnowska S, Knas M, Szajda S, et al. The activity of selected glycosidases in salivary gland tumors. Folia Histochem Cytobiol 2010;48:471–474. [37] Chojnowska S, Kępka A, Szajda SD, Waszkiewicz N, Bierć M, Zwierz K. Exoglycosidase markers of diseases. Biochem Soc Trans 2011;39:406–409. [38] Steinke J, Crouse C, Bradley D, Hise K, Lynch K, Kountakis SE, et al. Characterization of interleukin-4-stimulated nasal polyps fibroblasts. Am J Respir Cell Mol Biol 2004;30:212–219. [39] Xaubet A, Mullol J, Roca-Ferrer J, Pujols L, Fuentes M, Pérez M, et al. Effect of budesonide and nedocromil sodium on IL6 and IL-8 release from human nasal mucosa and polyp epithelial cells. Respir Med 2001;95:408–414. [40] Onerci M, Elsurer C, Guzel EE, Dagdeviren A. Distribution of inflammatory cells, adhesion molecules, intermediate filaments, and chemokine receptors in subgroups of nasal polyp patients. Am J Rhinol Allergy 2011;25:76–80. [41] Tinsgaard PK, Larsen PL, Bock T, Lange V, Tos M. Expression of intercellular adhesion molecule-1 on the vascular endothelium in nasal polyps before, during and after topical glucocorticoid treatment. Acta Otolaryngol (Stockh) 1998;118:404–408.
197
[42] Wang X, Gong S. Effects of corticosteroid on Eotaxin and Eotaxin-2 in nasal polyps. Lin Chung Er Bi Yan Hou Tou Jing Wai Ke Za Zhi 2009;23(5):205–208. [43] Zhang G, Shao J, Su C, Zhao X, Wang X, Sun X, et al. Distribution change of mast cells in human nasal polyps. Anat Rec (Hoboken) 2012. http://dx.doi.org/10.1002/ar.22430 [Epub ahead of print]. [44] Pletcher SD, Goldberg AN. Treatment of recurrent sinonasal polyposis with steroid-infused carboxymethylcellulose foam. Am J Rhinol Allergy 2010;24:451–453. [45] Rostkowska-Nadolska B, Preś K, Frączkowska K. Nasal water spray of 22R-16a,17a-butylidenodioxy-11b,21dihydroxy-1,4-pregnadien-3,20-dione (22R-bdpd) in prevention and treatment of nasal polyposis. Adv Clin Exp Med 2005;14:497–503. [46] Metzler EO, Hamilos DL, Hadley JA, Lanza DC, Marple BF, Nicklas RA, et al. Rhinosinusitis: establishing definitions for clinical research and patient care. Otolaryngol Head Neck Surg 2004;131:1–62. [47] Sitarek P, Zielińska-Bliźniewska H, Miłoński J, Przybyłowska K, Majsterek I, Olszewski J. Role of the 765 G/C polymorphism of COX-2 gene in pathogenesis of chronic rhinosinusitis with nose polyps in a Polish population. Otolaryngol Pol 2012;66:181–184. [48] Bernstein JM, Yankaskas JR. Increased ion transport in cultured nasal epithelial cells. Arch Otolaryngol Head Neck Surg 1994;120:993–996. [49] Bernstein JM, Yankaskas JR. Electrolyte and water transport and biological properties of nasal polyps. In: Red Mygind N, Lildhaldt T, editors. W: nasal polyp an inflammatory disease its treatment. Munsgaard: Copenhagen; 1997. p. 44–49. [50] Wasiluk A, Waszkiewicz N, Szajda SD, WojewódzkaŻelezniakowicz M, Kępka A, Minarowska A, et al. Alpha fucosidase and beta galactosidase in serum of a Lyme disease patients as a possible marker of accelerated senescence – a preliminary study. Folia Histochem Cytobiol 2012;50:147–157. [51] Hardingham TE, Perkins J, Muir H. Molecular conformations in proteoglycan aggregation. Biochem Soc Trans 1983;11:128–130. [52] Tomaszewski JJ, Donica H. Structure and functions of proteoglycans. Post Biochem 1988;34:209–228.
