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Roomans, Godfried
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Publications (10 of 20) Show all publications
Servetnyk, Z. & Roomans, G. M. (2007). Chloride transport in NCL-SG3 sweat gland cells: channels involved. Experimental and molecular pathology (Print), 83(1), 47-53
Open this publication in new window or tab >>Chloride transport in NCL-SG3 sweat gland cells: channels involved
2007 (English)In: Experimental and molecular pathology (Print), ISSN 0014-4800, E-ISSN 1096-0945, Vol. 83, no 1, p. 47-53Article in journal (Refereed) Published
Abstract [en]

The aim of the study was to assess whether NCL-SG3, the only immortalized sweat gland cell line available, can be used as an in vitro model to study chloride ion transport in cultured sweat gland cells. Cl efflux was measured using the MQAE dye fluorescence technique after stimulating the cells with different agonists. A significant stimulation of chloride efflux was achieved with the calcium ionophore A23187 resulting in an efflux rate of 0.9 mM/s. Both ATP and UTP activated chloride efflux in these cells, with the ATP response being larger. IBMX and forskolin stimulation did not induce a rate of chloride efflux above the basal level. Immunocytochemistry showed no detectable CFTR in NCL-SG3 cells. This finding was confirmed with flow cytometry analysis. Niflumic acid (20 and 100 μM NFA) and 4,4′-diisothiocyanatodihydrostilbene-2,2′-disulfonic acid (H2DIDS) (100 ìM) decreased the rate of ATP-stimulated chloride efflux significantly (0.40 and 0.31 mM/s with NFA, 0.37 mM/s with H2DIDS). Gadolinium (20 ìM) had no effect on the chloride transport rate. In conclusion, the NCL-SG3 cells retain some of the aspects of human sweat gland epithelium, such as the ability to form cell–cell contacts. The CFTR protein is neither functional nor expressed in cultured NCL-SG3 sweat gland cells. Ca2+-activated chloride conductance is confirmed and the putative Ca2+-activated chloride channel (CaCC) is further characterized in term of its pharmacological sensitivity. The NCL-SG3 sweat gland cell line can be used to investigate the characteristics of the CaCC and to identify the channel.

Keywords
Biological Markers, Calcium/metabolism, Cell Line, Chloride Channels/*metabolism, Chlorides/*metabolism, Epithelial Cells/metabolism, Humans, Immunohistochemistry, Ion Channel Gating, Ion Transport, Microscopy; Electron; Transmission, Sweat Glands/*metabolism/ultrastructure
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-17161 (URN)10.1016/j.yexmp.2007.02.003 (DOI)000247716800008 ()17383636 (PubMedID)
Available from: 2008-06-17 Created: 2008-06-17 Last updated: 2017-12-08Bibliographically approved
Nilsson, H., Dragomir, A., Ahlander, A., Johannesson, M. & Roomans, G. M. (2007). Effects of hyperosmotic stress on cultured airway epithelial cells. Cell and Tissue Research, 330(2), 257-269
Open this publication in new window or tab >>Effects of hyperosmotic stress on cultured airway epithelial cells
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2007 (English)In: Cell and Tissue Research, ISSN 0302-766X, E-ISSN 1432-0878, Vol. 330, no 2, p. 257-269Article in journal (Refereed) Published
Abstract [en]

Inhalation of hyperosmotic solutions (salt, mannitol) has been used in the treatment of patients with cystic fibrosis or asthma, but the mechanism behind the effect of hyperosmotic solutions is unclear. The relation between osmolarity and permeability changes was examined in an airway cell line by the addition of NaCl, NaBr, LiCl, mannitol, or xylitol (295–700 mOsm). Transepithelial resistance was measured as an indicator of the tightness of the cultures. Cell-cell contacts and morphology were investigated by immunofluorescence and by transmission electron microscopy, with lanthanum nitrate added to the luminal side of the epithelium to investigate tight junction permeability. The electrolyte solutions caused a significant decrease in transepithelial resistance from 450 mOsm upwards, when the hyperosmolar exposure was gradually increased from 295 to 700 mOsm; whereas the nonelectrolyte solutions caused a decrease in transepithelial resistance from 700 mOsm upwards. Old cultures reacted in a more rigid way compared to young cultures. Immuno-fluorescence pictures showed weaker staining for the proteins ZO-1, claudin-4, and plakoglobin in treated samples compared to the control. The ultrastructure revealed an increased number of open tight junctions as well as a disturbed morphology with increasing osmolarity, and electrolyte solutions opened a larger proportion of tight junctions than nonelectrolyte solutions. This study shows that hyperosmotic solutions cause the opening of tight junctions, which may increase the permeability of the paracellular pathway and result in increased transepithelial water transport.

Keywords
Airway epithelial cells, Hypertonic conditions, Permeability, Tight junctions, Transepithelial resistance, Cell culture
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-13920 (URN)10.1007/s00441-007-0482-7 (DOI)000249917000006 ()17768643 (PubMedID)
Available from: 2008-01-28 Created: 2008-01-28 Last updated: 2017-12-11Bibliographically approved
Nilsson, H., Dragomir, A., Ahlander, A., Ljungkvist, M. & Roomans, G. M. (2006). A modified technique for the impregnation of lanthanum tracer to study the integrity of tight junctions on cells grown on a permeable substrate. Microscopy research and technique (Print), 69(10), 776-783
Open this publication in new window or tab >>A modified technique for the impregnation of lanthanum tracer to study the integrity of tight junctions on cells grown on a permeable substrate
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2006 (English)In: Microscopy research and technique (Print), ISSN 1059-910X, E-ISSN 1097-0029, Vol. 69, no 10, p. 776-783Article in journal (Refereed) Published
Abstract [en]

Ionic lanthanum is commonly used to trace permeability pathways across epithelia and endothelia in biological electron microscopy. A method for obtaining a uniformly dense precipitate of lanthanum is described. The method, which is a modification of the technique described by Shaklai and Tavassoli (1977) was suitable for fixation of cell cultures grown on permeable filter inserts and was successfully applied to study opening of tight junctions by hypertonic solutions in the airway epithelial cell line 16HBE14o(-). The preparation method formed the basis for a semi-quantitative morphological determination in which the tight junctions were subdivided as "intact," "weakened," and "open." By using this modified technique, it could be demonstrated that opening of tight junctions in airway epithelial cells increased, with increasing osmolarity with electrolytes having a stronger effect than nonelectrolytes. A significant linear relationship was found between the osmolarity of the medium and the open state of the tight junctions (as determined by the semi-quantitative morphological technique) or the transepithelial electrical resistance.

Keywords
lanthanum impregnation, airway epithelial cells, permeable substrate, tight junctions, hypertonic conditions, transepithelial resistance, permeability
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-24744 (URN)10.1002/jemt.20347 (DOI)000241261400002 ()16865714 (PubMedID)
Available from: 2007-02-07 Created: 2007-02-07 Last updated: 2017-12-07Bibliographically approved
Servetnyk, Z., Krjukova, J., Gaston, B., Zaman, K., Hjelte, L., Roomans, G. M. & Dragomir, A. (2006). Activation of chloride transport in CF airway epithelial cell lines and primary CF nasal epithelial cells by S-nitrosoglutathione. Respiratory research (Online), 7, 124
Open this publication in new window or tab >>Activation of chloride transport in CF airway epithelial cell lines and primary CF nasal epithelial cells by S-nitrosoglutathione
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2006 (English)In: Respiratory research (Online), ISSN 1465-9921, E-ISSN 1465-993X, Vol. 7, p. 124-Article in journal (Refereed) Published
Abstract [en]

Background: It has been suggested that low mu M concentrations of S-nitrosoglutathione (GSNO), an endogenous bronchodilator, may promote maturation of the defective cystic fibrosis (CF) transmembrane conductance regulator ( CFTR). Because nitric oxide ( NO) and GSNO levels appear to be low in the CF airway, there is an interest in the possibility that GSNO replacement could be of therapeutic benefit in CF.

Methods: The effect of GSNO on chloride (Cl-) transport was investigated in primary nasal epithelial cells obtained from CF patients homozygous for the delF508 mutation, as well as in two CF cell lines (CFBE and CFSME), using both a fluorescent Cl- indicator and X-ray microanalysis. Maturation of delF508 CFTR was determined by immunoblotting.

Results: Treatment with 60 mu M GSNO for 4 hours increased cAMP-induced chloride efflux in nasal epithelial cells from 18 out of 21 CF patients, but did not significantly affect Cl- efflux in cells from healthy controls. This Cl- efflux was confirmed by measurements with a fluorescent Cl- indicator in the CFBE and CFSME cell lines. The effect of GSNO on Cl- efflux in CFBE cells could be inhibited both by a specific thiazolidinone CFTR inhibitor (CFTRinh-172) and by 4,4'-diisothiocyanatodihydrostilbene- 2,2'-disulfonic acid (H2DIDS). X-ray microanalysis showed that, following 4 hours incubation with 60 mu M GSNO, cAMP agonists caused a decrease in the cellular Cl- concentration in CFBE cells, corresponding to Cl- efflux. GSNO exposure resulted in an increase in the protein expression and maturation, as shown by immunoblot analysis. GSNO did not increase the cytosolic Ca2+ concentration in cultured airway epithelial cells.

Conclusion: Previous studies have suggested that treatment with GSNO promotes maturation of delF508-CFTR, consistent with our results in this study. Here we show that GSNO increases chloride efflux, both in the two CF cell lines and in primary nasal epithelial cells from delF508-CF patients. This effect is at least in part mediated by CFTR. GSNO may be a candidate for pharmacological treatment of the defective chloride transport in CF epithelial cells.

National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-24748 (URN)10.1186/1465-9921-7-124 (DOI)000241188400001 ()17022806 (PubMedID)
Available from: 2007-02-07 Created: 2007-02-07 Last updated: 2017-12-07Bibliographically approved
Vanthanouvong, V., Kozlova, I., Johannesson, M., Nääs, E., Nordvall, L., Dragomir, A. & Roomans, G. M. (2006). Composition of nasal airway surface liquid in cystic fibrosis and other airway diseases determined by X-ray microanalysis.. Microsc Res Tech, 69(4), 271-6
Open this publication in new window or tab >>Composition of nasal airway surface liquid in cystic fibrosis and other airway diseases determined by X-ray microanalysis.
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2006 (English)In: Microsc Res Tech, ISSN 1059-910X, Vol. 69, no 4, p. 271-6Article in journal (Refereed) Published
Keywords
Adolescent, Adult, Body Fluids/*chemistry, Child, Chlorine/analysis, Cystic Fibrosis/*metabolism, Electron Probe Microanalysis, Female, Heterozygote, Humans, Kartagener Syndrome/*metabolism, Male, Middle Aged, Nasal Lavage Fluid/*chemistry, Nasal Mucosa/*chemistry, Potassium/analysis, Research Support; Non-U.S. Gov't, Rhinitis/*metabolism, Salts/analysis, Sex Factors, Sodium/analysis
Identifiers
urn:nbn:se:uu:diva-19185 (URN)16586482 (PubMedID)
Available from: 2007-02-28 Created: 2007-02-28 Last updated: 2011-01-11
Shahana, S., Jaunmuktane, Z., Stenkvist Asplund, M. & Roomans, G. M. (2006). Ultrastructural investigation of epithelial damage in asthmatic and non-asthmatic nasal polyps. Respiratory Medicine, 100(11), 2018-2028
Open this publication in new window or tab >>Ultrastructural investigation of epithelial damage in asthmatic and non-asthmatic nasal polyps
2006 (English)In: Respiratory Medicine, ISSN 0954-6111, E-ISSN 1532-3064, Vol. 100, no 11, p. 2018-2028Article in journal (Refereed) Published
Abstract [en]

Nasal polyposis is a poorly understood chronic inflammatory disease often associated with asthma. As nasal polyps and asthma both are associated with massive eosinophil infiltration, they may share a common pathophysiological mechanism. Many genetic and autoimmune diseases may result from altered expression or function of cell adhesion molecules such as desmosomes. A transmission electron microscopical study was carried out on tissue from 15 patients suffering from nasal polyps, to investigate if there are changes in desmosomes in nasal polyps from asthmatic and/or allergic patients versus non-asthmatic versus non-allergic patients. In allergic patients the damage to columnar cells was more extensive than in non-allergic patients. Massive infiltration of eosinophils was observed in epithelium and connective tissue in all groups. No significant difference in thickness of the basal lamina was found between any of the groups. All patients had dilated capillaries in the connective tissue. The intercellular space between the epithelial cells was smallest in the asthmatic non-allergic group. The relative length of columnar cell or basal cell desmosomes was reduced in patients with asthma or allergy, compared to non-allergic, non-asthmatic patients. Hence, there appears to be a weakness in the desmosomes in asthmatics and allergics. Epithelial shedding may play an important rote in the pathophysiological process of a multifactorial disease such as asthma.

Keywords
nasal polyposis, asthma, epithelial damage, desmosomes, eosinophils
National Category
Medical and Health Sciences
Identifiers
urn:nbn:se:uu:diva-92579 (URN)10.1016/j.rmed.2006.02.012 (DOI)000241709900020 ()16580832 (PubMedID)
Available from: 2005-02-14 Created: 2005-02-14 Last updated: 2017-12-14Bibliographically approved
Kozlova, I., Nilsson, H., Henriksnäs, J. & Roomans, G. M. (2006). X-ray microanalysis of apical fluid in cystic fibrosis airway epithelial cell lines.. Cell Physiol Biochem, 17(1-2), 13-20
Open this publication in new window or tab >>X-ray microanalysis of apical fluid in cystic fibrosis airway epithelial cell lines.
2006 (English)In: Cell Physiol Biochem, ISSN 1015-8987, Vol. 17, no 1-2, p. 13-20Article in journal (Refereed) Published
Keywords
Body Fluids/chemistry, Cell Line, Cystic Fibrosis/*metabolism, Electron Probe Microanalysis, Epithelial Cells/metabolism, Humans, Hydrogen-Ion Concentration, Microscopy; Electron, Respiratory System/*metabolism
Identifiers
urn:nbn:se:uu:diva-24742 (URN)16543717 (PubMedID)
Available from: 2007-03-06 Created: 2007-03-06 Last updated: 2011-01-11
Relova, A.-J., Shahana, S., Makeeva, N. & Roomans, G. M. (2005). Effect of cytokines on ICAM-1 and ZO-1 expression on human airway epithelial cells.. Cell Biol Int, 29(9), 768-77
Open this publication in new window or tab >>Effect of cytokines on ICAM-1 and ZO-1 expression on human airway epithelial cells.
2005 (English)In: Cell Biol Int, ISSN 1065-6995, Vol. 29, no 9, p. 768-77Article in journal (Refereed) Published
Identifiers
urn:nbn:se:uu:diva-79752 (URN)16087364 (PubMedID)
Available from: 2006-04-13 Created: 2006-04-13 Last updated: 2011-01-11
Kozlova, I., Vanthanouvong, V., Almgren, B., Högman, M. & Roomans, G. M. (2005). Elemental composition of airway surface liquid in the pig determined by x-ray microanalysis.. Am J Respir Cell Mol Biol, 32(1), 59-64
Open this publication in new window or tab >>Elemental composition of airway surface liquid in the pig determined by x-ray microanalysis.
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2005 (English)In: Am J Respir Cell Mol Biol, ISSN 1044-1549, Vol. 32, no 1, p. 59-64Article in journal (Refereed) Published
Keywords
Animals, Bronchoalveolar Lavage Fluid/*chemistry, Chlorides/metabolism, Comparative Study, Electron Probe Microanalysis, Epithelium/*metabolism/ultrastructure, Ions/*metabolism, Microscopy; Electron; Transmission, Plasma/*chemistry, Potassium/metabolism, Research Support; Non-U.S. Gov't, Respiratory System/*metabolism/ultrastructure, Sodium/metabolism, Swine
Identifiers
urn:nbn:se:uu:diva-79756 (URN)15489275 (PubMedID)
Available from: 2006-04-13 Created: 2006-04-13 Last updated: 2011-01-11
Vanthanouvong, V., Kozlova, I. & Roomans, G. M. (2005). Ionic composition of rat airway surface liquid determined by X-ray microanalysis.. Microsc Res Tech, 68(1), 6-12
Open this publication in new window or tab >>Ionic composition of rat airway surface liquid determined by X-ray microanalysis.
2005 (English)In: Microsc Res Tech, ISSN 1059-910X, Vol. 68, no 1, p. 6-12Article in journal (Refereed) Published
Keywords
Animals, Body Fluids/*chemistry, Electron Probe Microanalysis, Hypotonic Solutions/chemistry, Ions/*analysis, Mucus/*chemistry, Nasal Mucosa/chemistry, Phosphorus/analysis, Potassium/analysis, Rats, Research Support; Non-U.S. Gov't, Respiratory Mucosa/*chemistry, Sodium/analysis, Sulfur/analysis, Trachea/chemistry
Identifiers
urn:nbn:se:uu:diva-79751 (URN)16208720 (PubMedID)
Available from: 2006-04-13 Created: 2006-04-13 Last updated: 2011-01-11
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