Extracts of different pollen species and their effect on human tear fluid and an epithelial cell line.

PURPOSE: Hazelnut and birch pollen are known to destroy tear film components and attack ocular surface cells. We investigated further pollen species from different plant families, whether they show similar effects on human tear fluid and an epithelial cell line in vitro, to provide a broad basis for further research on pollen reactions affecting the tear film and ocular surface. MATERIALS AND METHODS: Regional pollen species from different plant families (Adoxaceae, Betulaceae, Fagaceae, Juglandaceae, Malvaceae, Oleaceae, Pinaceae, Plantaginaceae, Poaceae, Salicaceae, Sapindaceae) were collected. Their proteolytic activity was evaluated by Zymography. Human tear fluid and cells of an epithelial cell line were incubated with pollen extracts. Tear fluid was analyzed by Polyacrylamide gel electrophoresis (PAGE). Cytomorphology was assessed microscopically and cell viability by proliferation (MTS), water-soluble tetrazolium (WST-1) assay and the impedance-based xCELLigence real-time analysis (RTCA). RESULTS: Zymography revealed significant protease activity and PAGE showed the degradation of tear proteins by different pollen species. Cells incubated with pollen extracts presented dose- and time-dependent cytomorphological changes. MTS, WST-1, and RTCA revealed cytostatic as well as cytotoxic effects of pollen extracts. CONCLUSIONS: Pollen species from different plant families exert proteolytic activity and degrade human tear fluid as well as epithelial cells, which may play a crucial role in the pathogenesis of allergic and non-allergic reactions affecting the ocular surface.

Nasal mucus proteomic changes reflect altered immune responses and epithelial permeability in patients with allergic rhinitis.

BACKGROUND: Nasal mucus is the first-line defense barrier against (aero-) allergens. However, its proteome and function have not been clearly investigated. OBJECTIVE: The role of nasal mucus in the pathophysiology of allergic rhinitis was investigated by analyzing its proteome in patients with allergic rhinitis (n = 29) and healthy control subjects (n = 29). METHODS: Nasal mucus was collected with a suction device, tryptically digested, and analyzed by using liquid chromatography-tandem mass spectrometry. Proteins were identified by searching the SwissProt database and annotated by collecting gene ontology data from databases and existing literature. Gene enrichment analysis was performed by using Cytoscape/BINGO software tools. Proteins were quantified with spectral counting, and selected proteins were confirmed by means of Western blotting. RESULTS: In total, 267 proteins were identified, with 20 (7.5%) found exclusively in patients with allergic rhinitis and 25 (9.5%) found exclusively in healthy control subjects. Five proteins were found to be significantly upregulated in patients with allergic rhinitis (apolipoprotein A-2 [APOA2], 9.7-fold; α2-macroglobulin [A2M], 4.5-fold; apolipoprotein A-1 [APOA1], 3.2-fold; α1-antitrypsin [SERPINA1], 2.5-fold; and complement C3 [C3], 2.3-fold) and 5 were found to be downregulated (antileukoproteinase [SLPI], 0.6-fold; WAP 4-disulfide core domain protein [WFDC2], 0.5-fold; haptoglobin [HP], 0.7-fold; IgJ chain [IGJ], 0.7-fold; and Ig hc V-III region BRO, 0.8-fold) compared with levels seen in healthy control subjects. CONCLUSION: The allergic rhinitis mucus proteome shows an enhanced immune response in which apolipoproteins might play an important role. Furthermore, an imbalance between cysteine proteases and antiproteases could be seen, which negatively affects epithelial integrity on exposure to pollen protease activity. This reflects the important role of mucus as the first-line defense barrier against allergens.

Altered inhibitory function of the E-type prostanoid receptor 4 in eosinophils and monocytes from aspirin-intolerant patients.

Prostaglandin (PG) E2 has been implicated in the pathogenesis of aspirin-exacerbated respiratory disease (AERD). E-type prostanoid (EP) receptor 4 is known to confer inhibitory signals to eosinophils and monocytes, amongst others. In this study, we investigated whether the responsiveness of eosinophils and monocytes to PGE2 and EP4 receptor activation is altered in AERD patients. While the expression of the EP4 receptor in eosinophils was unaltered in AERD patients, inhibition of eosinophil chemotaxis by PGE2 or the EP4 agonist CAY10598 was less pronounced in AERD patients as compared to healthy control subjects. In monocytes, we found no changes in basal or lipopolysaccharide (LPS)-stimulated PGE2 synthesis, but the response to EP4 receptor activation with respect to inhibition of LPS-induced tumor necrosis factor-α release was reduced in AERD patients, especially in the presence of aspirin (acetylsalicylic acid). Our data point towards a decreased sensitivity of inhibitory EP4 receptor that may play a role in AERD.

EP4 receptor stimulation down-regulates human eosinophil function.

Accumulation of eosinophils in tissue is a hallmark of allergic inflammation. Here we observed that a selective agonist of the PGE(2) receptor EP4, ONO AE1-329, potently attenuated the chemotaxis of human peripheral blood eosinophils, upregulation of the adhesion molecule CD11b and the production of reactive oxygen species. These effects were accompanied by the inhibition of cytoskeletal rearrangement and Ca(2+) mobilization. The involvement of the EP4 receptor was substantiated by a selective EP4 antagonist, which reversed the inhibitory effects of PGE(2) and the EP4 agonist. Selective kinase inhibitors revealed that the inhibitory effect of EP4 stimulation on eosinophil migration depended upon activation of PI 3-kinase and PKC, but not cAMP. Finally, we found that EP4 receptors are expressed by human eosinophils, and are also present on infiltrating leukocytes in inflamed human nasal mucosa. These data indicate that EP4 agonists might be a novel therapeutic option in eosinophilic diseases.

Current and emerging pharmacotherapy for pediatric allergic rhinitis.

INTRODUCTION: Allergic rhinitis (AR) is a global health problem in adults as well as the younger population, continuously increasing and posing a significant problem for patients, health care systems and economies. For the younger population, some aspects differ from treatment of adults, namely, prevention, compliance and adherence. AREAS COVERED: This narrative review summarizes all the pharmacotherapeutic options with special focus on the pediatric population. Moreover, it elucidates prevention strategies as well as future developments of AR treatment. Currently, symptomatic therapy in the form of steroids and antihistamines is applied topically and systemically where steroids need to be administered with caution and for a very short term. The only disease-modifying and causal treatment is allergen immunotherapy administered sublingually and subcutaneously. Future and current novel therapeutic options are human monoclonal antibodies. EXPERT OPINION: The greatest potential for future developments currently lie in allergen immunotherapy and here in different routes of administration and modification of (recombinant) allergens as well as immune-modulating adjuvants and nanoparticles. Secondly, monoclonal antibodies are promising molecules blocking and/or interfering with up- and downstream immune mechanisms. Another important aspect lies in prevention of allergic sensitization and disease progression through both AIT and biologics which is particularly true for the pediatric population.

Olfactory cleft proteome does not reflect olfactory performance in patients with idiopathic and postinfectious olfactory disorder: A pilot study.

Technical advances including liquid chromatography-tandem mass spectrometry and its data analysis enable detailed proteomic analysis of the nasal mucus. Alterations of the nasal mucus proteome may provoke substantial changes of the nasal physiology and have already been associated with rhinologic diseases such as allergic rhinitis. This study was conducted as a pilot study to map the olfactory cleft proteome using current techniques for proteomic analysis. Furthermore, we aimed to investigate proteomic changes as potential biomarkers in patients suffering from idiopathic and postinfectious olfactory disorders compared to healthy controls. Seven patients with idiopathic hyposmia and anosmia, seven patients with postinfectious hyposmia and anosmia and seven healthy controls were included in this study. In total, 1117 different proteins were detected in at least five patients in at least one group. Results of this study did not reveal significant differences regarding the proteomic composition of the olfactory cleft mucus between patients versus healthy controls. Among proteins involved in olfactory perception the G protein family was detected but also found unchanged between groups. Investigation of protein composition by liquid chromatography-tandem mass spectrometry enabled us to perform an in-depth analysis of the olfactory cleft mucus proteome regarding the diversity of different proteins in individual patients. However untargeted proteomics of the olfactory cleft mucus may not be an applicable approach to develop biomarkers for olfactory disorders. Targeted analyses of distinct proteins known to be involved in olfactory perception but not detected by our approach, e.g. odorant binding proteins, may provide more information regarding pathophysiology of olfactory diseases.

Plasmapheresis reverses all side-effects of a cisplatin overdose--a case report and treatment recommendation.

BACKGROUND: Cisplatin is widely used as an antineoplastic agent since it is effective against a broad spectrum of different tumours. Nevertheless, it has several potential side effects affecting different organ systems and an overdose may lead to life-threatening complications and even death. CASE PRESENTATION: We report on a 46-year old woman with non-small cell lung cancer who accidentally received 225 mg/m2 of cisplatin, which was threefold the dose as scheduled, within a 3-day period. Two days later, the patient presented with hearing loss, severe nausea and vomiting, acute renal failure as well as elevated liver enzymes. In addition, she developed a severe myelodepression. After plasmapheresis on two consecutive days and vigorous supportive treatment, the toxicity-related symptoms improved and the patient recovered without any sequelae. CONCLUSION: To date, no general accepted guidelines for the treatment of cisplatin overdoses are available. Along with the experience from other published cases, our report shows that plasmapheresis is capable of lowering cisplatin plasma and serum levels efficiently. Therefore, plasma exchange performed as soon as possible can ameliorate all side effects of a cisplatin overdose and be a potential tool for clinicians for treatment. However, additional intensive supportive treatment-modalities are necessary to control all occurring side effects.

Seasonal proteome changes of nasal mucus reflect perennial inflammatory response and reduced defence mechanisms and plasticity in allergic rhinitis.

INTRODUCTION: Nasal mucus and its proteins are a defence against allergens. We sought to investigate dynamic proteome changes in allergic rhinitis upon environmental allergen provocation. METHODS: Nasal mucus was collected in and out of pollen season from allergic rhinitis patients (N=10) and healthy controls (N=12). Liquid chromatography-tandem mass spectrometry was performed. Proteins were identified by SwissProt database search and quantified from normalized areas under curve of precursor ion chromatograms. Gene enrichment analysis was performed with Cytoscape/BINGO software. RESULTS: In total 430 different proteins were detected in both groups, 203 (47.2%) were newly identified. In allergics CLU and IGKC were significantly more abundant in season (2.2 and 2.1-fold respectively). GSTP1 (0.5-fold), ELANE (0.4-fold), HIST1H2BK (0.3-fold), S100A8 (0.2-fold), S100A12 (0.2-fold) and ARHGDIB (0.1-fold) were significantly less abundant in season. In healthy controls UBC, TUBA1B, HBB and FABP5 were only present in season. Ig kappa chain V-I region DEE (5.3-fold), CLU (5.0-fold), TXN (4.3-fold), MSMB (3.2-fold) and Ig heavy chain V-III region BRO (2.7-fold) were significantly more abundant in season. MUC5B (0.5-fold), SLPI (0.2-fold) and S100P (0.2-fold) were significantly less abundant in season. CONCLUSION: Contrary to their symptoms allergic rhinitis patients show perennial inflammatory response lacking adequate reaction to allergens in season. BIOLOGICAL SIGNIFICANCE: Many studies dealing with allergic rhinitis are focused on the nasal epithelium. This is the first study to analyse the nasal mucus as primary defence barrier on a proteomic level in and out of pollen season and contrary to the leading opinion shows that allergic patients show a perennial inflammatory response with reduced reaction to allergens whereas healthy controls react on proteome basis towards enhanced defence in season despite lacking allergic sensitization.

Potential correlations of dentogenic factors to the development of clinically verified fungus balls: A retrospective computed tomography-based analysis.

OBJECTIVES/HYPOTHESIS: Fungus balls are a common disease of the paranasal sinuses, usually involving the maxillary sinus. To clarify the pathology, we analyzed patients treated for maxillary sinus fungus balls to see whether the latter correlated with dentogenic factors. STUDY DESIGN: Retrospective case analysis. METHODS: Cases of maxillary sinus fungus balls diagnosed between January 2000 and December 2013 were analyzed retrospectively. Patients' charts were reviewed for diagnosis, gender, and age. Paranasal sinus computed tomography (CT) scans were reviewed according to the side of the fungus ball, calcifications/opacifications, and dentogenic factors. RESULTS: In 98/102 patients (96.1%), 157 dentogenic factors could be identified on the side affected by a fungus ball. On the contralateral healthy side, there were 125 dentogenic factors. In four (3.9%) of the patients, no dentogenic pathology was identified on the CT scan. The presence of dentogenic factors (regardless of number) was significantly associated with a fungus ball compared to the healthy side (P = .024, χ(2) test, odds ratio: 2.72 [95% confidence interval: 1.02-7.23]). CONCLUSIONS: Dentogenic factors regardless of type potentially correlate with the presence of maxillary sinus fungus ball. Unlike the overall presence of dentogenic factors, the particular dentogenic factors in an individual patient do not significantly influence the development of fungus balls. After diagnosis of dentogenic pathology in penetrated maxillary sinus floors, patients should be closely monitored and informed about their higher risk of developing a fungus ball.

Apolipoproteins have a potential role in nasal mucus of allergic rhinitis patients: a proteomic study.

OBJECTIVES/HYPOTHESIS: Nasal mucus is a defense barrier against aeroallergens. We recently found apolipoproteins to be elevated in the nasal mucus of allergic rhinitis patients. Apolipoproteins are involved in lipid metabolism, have immunomodulatory properties, and may represent interesting novel biomarkers. This study aims to validate our findings and analyze whether the increased abundance of apolipoproteins in nasal mucus is a local or systemic phenomenon in allergic rhinitis. STUDY DESIGN: Prospective controlled trial. METHODS: Nasal mucus of allergic rhinitis patients (n = 10) and healthy controls (n = 12) was collected, tryptically digested, and analyzed by LC-MS/MS. Areas under the curve (AUCs) of the total peptides identified and matched to apolipoproteins were used to compare relative protein abundances of the same protein between groups. RESULTS: In a total of 389 identified proteins in nasal mucus, apolipoproteins A-I, A-II, A-IV, and B 100 were detected. Apolipoprotein A-I (mean normalized AUC 1.49% [SEM = 0.5] vs. 0.42% [SEM = 0.2]) and A-II (mean normalized AUC 0.47% [SEM = 0.2] vs. 0.05% [SEM = 0.02]) were significantly more abundant in allergic rhinitis patients than controls (3.6-fold and 9.4-fold, respectively). Apolipoprotein A-IV (mean normalized AUC = 0.01%) and B-100 (mean normalized AUC = 0.02%) were each detected in only one allergic rhinitis patient out of 10. Myeloperoxidase was detected with a mean normalized AUC of 0.06% (SEM = 0.03) in allergic rhinitis patients and 0.18% (SEM = 0.08) in healthy controls without reaching significance. CONCLUSION: This study confirms the significantly higher abundance of apolipoproteins A-I and AII in allergic rhinitis mucus. Their release seems to be triggered by local mechanisms as an antiinflammatory response to allergens.

Influence of sensitization to inhalative allergens on adenotonsillar disease.

OBJECTIVE: We sought to determine the influence of IgE-mediated sensitization on adenotonsillar disease in children. We compared follow-up after tonsillectomy/adenoidectomy of atopic and nonatopic children.Study design and setting A prospective study of 293 children consecutively undergoing tonsillectomy/adenoidectomy was conducted at a university hospital center. Preoperative and postoperative (1-year follow-up) allergy-related symptom scores were obtained by parents. Intraoperative total serum IgE, the screening test sx1 (Pharmacia), and, if positive, serum specific IgE to inhalative allergens were carried out. RESULTS: Sixty-seven children (22.9%) showed positive RAST results to inhalative allergens (class 1 to 6). In both sensitized and nonsensitized groups, the general health was improved in more than 89% at 1 year postoperatively. No significant difference of postoperative nasal symptoms between the atopic and nonatopic groups was found. CONCLUSION: Both atopic and nonatopic children with adenotonsillar disease improve health after adenoidectomy or adenotonsillectomy. In our opinion, routine allergy tests before adenotonsillectomy or adenoidectomy are not justified. Only if allergy is suspected due to clinical findings or family history should an allergy test be carried out.

Feasibility of routine paranasal sinus CT-scans in preoperative transplant patients.

BACKGROUND: Paranasal Sinus (PNS) CT- scans are routinely performed for screening of sinusitis in patients scheduled for organ transplant at the University Hospital Graz. This study was performed to evaluate whether routine PNS CT-scans are necessary in every transplant patient or in selected cases only, because clear evidence is missing in literature. MATERIAL/METHODS: 142 patients (113 male, 29 female) were included in this retrospective patient chart study. RESULTS: The majority of patients (n=142) were scheduled for liver transplant (n=79). Of all patients (n=142), 50 (35.2%) had normal PNS CT-scans and 92 (64.8%) had a pathologically changed PNS-scan. Seventy-one out of these 92 (77.2%) patients did not show clinical symptoms of sinusitis nor changes in diagnostic endoscopy, 21 (22.8%) patients did have symptoms of sinusitis and diseased PNS CT-scans. Three out of 71 (4.2%) without symptoms but pathologic CT-scans compared to 7/21 (33.3%) patients with symptoms and pathological CT-scans underwent endoscopic paranasal sinus surgery (FESS). All patients operated suffered from chronic sinusitis, none had mycotic or neoplastic disease in definite histology. CONCLUSIONS: Since there was a high rate (77.2%) of pathological CT scans unrelated to sinusitis symptoms these findings should be considered as accidental. In our opinion routine paranasal sinus CT scans are not feasible in patients scheduled for organ transplant according to high rate of false positive results.

Nasal IL-5 levels determine the response to anti-IL-5 treatment in patients with nasal polyps.

BACKGROUND: Chronic rhinosinusitis with nasal polyps is characterized by an eosinophilic inflammation and high IL-5 levels. OBJECTIVES: Antagonizing the effect of IL-5 is a potential new treatment strategy in patients with nasal polyps. METHODS: In a double-blind, placebo-controlled, randomized, 2-center safety and pharmacokinetic study, 24 subjects with bilateral nasal polyps were randomized to receive a single intravenous infusion of reslizumab, a humanized anti-human IL-5 mAb, at 3 mg/kg or 1 mg/kg or placebo. We evaluated the safety and pharmacokinetics of reslizumab, and biologic activity was assessed by means of endoscopic evaluation of polyp size, symptoms, peripheral eosinophil counts, peripheral and local IL-5 levels, eotaxin levels, and eosinophil cationic protein levels. RESULTS: We demonstrated that a single injection of reslizumab up to 3 mg/kg is safe and well tolerated. Blood eosinophil numbers and concentrations of eosinophil cationic protein were reduced up to 8 weeks after treatment in serum and nasal secretions. Individual nasal polyp scores improved only in half of the treated patients for 4 weeks. Responders had increased IL-5 concentrations in nasal secretions at baseline compared with nonresponders, and logistic regression analysis revealed that increased nasal IL-5 levels (>40 pg/mL) predict the response to anti-IL-5 treatment. CONCLUSION: A single injection of anti-IL-5 reduces the size of nasal polyps for 4 weeks in half of the patients, and nasal IL-5 levels predict the response to anti-IL-5 treatment. CLINICAL IMPLICATIONS: Intravenous administration of a humanized anti-human IL-5 mAb is safe and reduces the size of nasal polyps in half of the patients.

5-Oxo-6,8,11,14-eicosatetraenoic acid is a potent chemoattractant for human basophils.

BACKGROUND: 5-Oxo-6,8,11,14-eicosatetraenoic acid (5-oxo-ETE) is a chemoattractant for eosinophils and neutrophils, and the messenger RNA for its receptor, the oxo-eicosatetraenoic acid receptor (OXE), has been detected in several tissues. OBJECTIVES: This study aimed at clarifying the role of 5-oxo-ETE in the regulation of basophil function. METHODS: Basophil responses were determined in assays of flow-cytometric shape change, Ca(2+) flux, chemotaxis, and histamine release. Messenger RNA for OXE was detected by real-time PCR. RESULTS: We observed that human eosinophils were 3 to 10 times more sensitive to 5-oxo-ETE than neutrophils in flow-cytometric shape change and Ca(2+) flux assays, as estimated from the half-maximal responses of the cells. Basophils responded to 5-oxo-ETE in the shape change assay with a sensitivity similar to that of eosinophils. 5-Oxo-ETE was a weak inducer of Ca(2+) flux in basophils and did not cause histamine release but was a highly effective chemoattractant for basophils in the low nanomolar concentration range in a pertussis toxin-sensitive manner. In agreement with these functional studies, the messenger RNA for the 5-oxo-ETE receptor, OXE, was detectable in basophils as in monocytes, eosinophils, and neutrophils, but not in fibroblasts. Specimens from sinus mucosa, tonsils, and adenoids also contained detectable levels of messenger RNA for OXE. CONCLUSION: Our data suggest that 5-oxo-ETE is potentially involved in the regulation of basophil recruitment and might hence be a useful therapeutic target in atopic disease.