Smoking is associated with increased levels of extracellular peptidylarginine deiminase 2 (PAD2) in the lungs.

OBJECTIVES: Smoking is a well-established risk factor in rheumatoid arthritis (RA), and citrullination of self-antigens plays a pathogenic role in the majority of patients. Increased numbers of peptidylarginine deiminase 2 (PAD2)-containing macrophages have been demonstrated in bronchoalveolar lavage (BAL) fluid from smokers, but intracellularly located PAD cannot be responsible for citrullination of extracellular self-antigens. We aimed to establish a link between smoking and extracellular PAD2 in the lungs. METHODS: BAL fluid samples were obtained from 13 smokers and 11 nonsmoking controls. Total protein content and C-reactive protein (CRP) concentration were determined after separating cells from the samples. PAD2 content in cell-free BAL fluids was measured by means of a PAD2-specific sandwich ELISA. RESULTS: Significantly increased levels of soluble PAD2 were detected in cell-free BAL fluids from smokers as compared to non-smokers (p=0.018). The PAD2 content correlated with the overall CRP levels (p=0.009) and cell count (p=0.016). CONCLUSIONS: This first demonstration of increased levels of extracellular PAD2 in the lungs of smokers supports the hypothesis that smoking promotes extracellular citrullination of proteins. This may represent a pathological event upstream for the production of anti-citrullinated protein antibodies (ACPAs) among RA patients carrying HLA-molecules capable of binding citrullinated self-peptides.

Surfactant proteins A, B, C and D in the human nasal airway: associated with mucosal glands and ciliated epithelium but absent in fluid-phase secretions and mucus.

AIMS: To investigate the presence of surfactant protein (SP) A, B, C and D in nasal airways and to determine whether the proteins exert their main functions in nasal secretions or in the deeper layers of the nasal mucosa. METHODS: Volunteers were recruited from the Department of ENT Head and Neck Surgery, Odense University Hospital, Denmark. The study included 39 subjects. Nasal mucosal biopsies were analyzed by immunohistochemistry, and bronchoalveolar and nasal lavages, nasal brush biopsies and nasal mucus were analyzed for SP-A, -B, -C and -D by SDS-PAGE and Western blotting. The presence of SP-A and SP-D in the first three samplings were also analyzed by enzyme-linked immunosorbent assay. RESULTS: In nasal mucosal biopsies, SP-A, -B, -C and -D were all demonstrated in the serous acini of the submucosal glands and in the surface epithelium. SP-D was detected in nasal brush biopsies, whereas the other SPs were absent. Moreover, SP-A, -B, -C and -D were absent in nasal lavage and mucus. CONCLUSION: SP-A, -B, -C and -D exert their protective effect in the ductal epithelium of the submucosal glands rather than in nasal secretions and mucus. Further studies are required to clarify the functions of these proteins in nasal secretory pathways for understanding upper airway diseases.

Biochemical and Bioimaging Evidence of Cholesterol in Acquired Cholesteatoma.

OBJECTIVES: To quantify the barrier sterols and image the lipid structures in the matrix of acquired cholesteatoma and compare the distribution with that found in stratum corneum from normal skin, with the goal to resolve their potential influence on cholesteatoma growth. METHODS: High-performance thin-layer chromatography (HPTLC) was used to achieve a quantitative biochemical determination of the sterols. The intercellular lipids were visualized by Coherent Anti-Stokes Raman scattering (CARS) microscopy, which enables label-free imaging of the lipids in intact tissue samples. RESULTS: The results show that the total lipid content of the cholesteatoma matrix is similar to that of stratum corneum from skin and that the cholesteatoma matrix unquestionably contains cholesterol. The cholesterol content in the cholesteatoma matrix is increased by over 30% (w/w dry weight) compared to the control. The cholesterol sulfate content is below 1% of the total lipids in both the cholesteatoma and the control. Cholesterol ester was reduced by over 30% when compared to the control. CONCLUSIONS: The content of cholesterol in the cholesteatoma matrix is significantly different from that in stratum corneum from skin, and we confirm that the main structure of the cholesteatoma resembles very thick stratum corneum.

Promising results after balloon dilatation of the Eustachian tube for obstructive dysfunction.

INTRODUCTION: As the first ear, nose and throat department in Denmark, we introduced balloon dilatation of the Eustachian tube as a treatment of obstructive dysfunction in the summer of 2012. We present our preliminary experiences with this new treatment in adults. MATERIAL AND METHODS: Preoperatively, several different tests were performed including otomicroscopy, audiometry, tympanometry and Toynbee's test. The patients were classified as Class 1 (if they could make a pressure equalisation of the middle ear by a normal Valsalva's test), Class 2 (if they needed an extended Valsalva's test), Class 3 (if only a test with the Otovent could make air flow to the middle ear), and Class 4 (if no passage of the Eustachian tube could be achieved). Furthermore, the patients filled out questionnaires using a visual analogue scale (VAS). RESULTS: A total of 50 treatments were performed in 34 patients (16 patients had bilateral problems). Four patients (six Eustachian tubes) had intermittent problems, while 30 patients had chronic dysfunction. A significant effect of the treatment was documented when measuring both audiometry, tympanometry, Toynbee's test, classification of Eustachian tube dysfunction and VAS questionnaires. Some patients (e.g. patients with atelectatic ear drums) were not helped by the treatment. Among the first 40 treatments, 10% were observed to have acute otitis media post-operatively. DISCUSSION: The majority of the patients experienced a positive effect of the treatment. Our results are comparable to those of other similar studies. We regard this new treatment as very promising, but look forward to more research. FUNDING: not relevant. TRIAL REGISTRATION: not relevant.

Structural characterization and lipid composition of acquired cholesteatoma: a comparative study with normal skin.

HYPOTHESIS: The goal of this work is to characterize the morphology and lipid composition of acquired cholesteatoma. We hypothesize that constitutive lipid membranes are present in the cholesteatoma and resemble those found in human skin stratum corneum. METHODS: We performed a comparative noninvasive structural and lipid compositional study of acquired cholesteatoma and control human skin using multiphoton excitation fluorescence microscopy-related techniques and high-performance thin-layer chromatography. RESULTS: The structural arrangement of the cholesteatoma is morphologically invariant along a depth of more than 200 µm and resembles the stratum corneum of hyperorthokeratotic skin. Lipid compositional analyses of the cholesteatoma show the presence of all major lipid classes found in normal skin stratum corneum (ceramides, long chain fatty acids, and cholesterol). Consistent with this, evaluation of Nile red and LAURDAN generalized polarization function images of the cholesteatoma show intercellular regions similar to normal skin stratum corneum in terms of lipid membrane packing and local water content. CONCLUSION: The investigations show the presence of an extremely thickened stratum corneum within the cholesteatoma. The lipid composition and extracellular membranes similar to those of normal skin stratum corneum are present, indicating that a defensive/permeability barrier is present in the cholesteatoma. Finally, it is demonstrated that multiphoton excitation fluorescence microscopy is a suitable noninvasive tool for investigating the morphology and intrinsic physical properties of acquired cholesteatoma.

Complement defects in patients with chronic rhinosinusitis.

The complement system is an important part of our immune system, and complement defects lead generally to increased susceptibility to infections and autoimmune diseases. We have studied the role of complement activity in relation with chronic rhinosinusitis (CRS), and more specifically studied whether complement defects collectively predispose individuals for CRS or affect CRS severity. The participants comprised 87 CRS patients randomly selected from the general population, and a control group of 150 healthy blood donors. The CRS patients were diagnosed according to the European Position Paper on Rhinosinusitis and nasal Polyps criteria, and severity was evaluated by the Sino-nasal Outcome Test-22. Serum samples were analysed by ELISA for activity of the respective pathways of complement, and subsequently for serum levels of relevant components. We found that the frequency of complement defects was significantly higher among CRS patients than among healthy control subjects. A majority of Mannan-binding lectin deficient CRS patients was observed. The presence of complement defects had no influence on the severity of subjective symptoms. Our studies show that defects in the complement system collectively may play an immunological role related to the development of CRS. However, an association between severity of symptoms and presence of complement defects could not be demonstrated.

[Meningioma as the cause of ear problems]. / Meningiom som årsag til øregener.

A 45-year-old woman with long-term unilateral ear problems, hearing loss, vertigo and tinnitus was diagnosed with a meningioma in the middle ear. Magnetic resonance imagining (MRI) revealed that the meningioma penetrated the petrous bone into the middle ear. The meningioma was removed to avoid further hearing loss. Patients with long-term unilateral ear problems that are resistant to conventional therapy should be referred to computed tomography, MRI and explorative tympanotomy. Surgery is indicated if progressions in neurological deficits are seen, but surgery may cause serious neurological deficits.

The distribution of free calcium ions in the cholesteatoma epithelium.

The distribution of free calcium ions in normal skin and cholesteatoma epithelium was investigated using the oxalate precipitation method. In agreement with previous observations, we could demonstrate a calcium ion gradient in normal epidermis where the cells in stratum basale and spinosum reside in an environment containing relatively low concentrations of calcium ions, whereas the outer stratum granulosum contained abundant calcium. The concentration declined precipitously in the stratum corneum. In contrast, in cholesteatomas, the gradient was perturbed in some areas of the nucleated layers and areas appeared where oblong accumulations of free calcium ions were found basally in the stratum. These findings provide evidence that fluctuations in epidermal calcium in cholesteatoma epithelium may underlie the abnormal desquamation, may contribute to the formation of an abnormal permeability barrier and may regulate terminal events in epidermal differentiation.

Altered permeability barrier structure in cholesteatoma matrix.

The stratum corneum of the cholesteatoma epithelium comprises the greater part of the cholesteatoma matrix. The permeability barrier that militates against diffusion and penetration of infectious and toxic agents into and through the epithelium is situated here. The multiple long sheets of lamellar lipid structures filling the intercellular spaces mainly control the barrier function. The barrier in cholesteatoma epithelium is several times thicker than in unaffected skin but presents distinctive features of a defective barrier as seen in other scaling skin diseases. The intercellular spaces appear rather twisted, and the extra-cellular barrier is not found until well above the stratum granulosum/stratum corneum interface; thus, the transformation from secreted Odland-body contents is disturbed. Large dilatations of the extra-cellular space that are sometimes filled with an electron-dense material frequently occur. The corneocytes are shed in clusters, not as single cells. Further, lipid droplets and intracellular membranous material are occasionally seen. In spite of these clear signs of barrier dysfunction, it is unknown whether the thickness of the barrier compensates for the defect in barrier efficiency. Despite this, one cannot determine whether the stimulus to the increased lipid metabolic activity originates from the middle ear or from the ear canal.