Search

Natalia Celejewska-Wójcik,1 Aleksander Kania,1 Karolina Górka,1 Paweł Nastałek,1 Krzysztof Wójcik,2 Anna Gielicz,3 Lucyna Mastalerz,1 Marek Sanak,3 Krzysztof Sładek1 1Department of Pulmonology, 2nd Department of Internal Medicine, Faculty of Medicine, Jagiellonian University Medical College, Kraków, Poland; 2 2nd Department of Internal Medicine, Faculty of Medicine, Jagiellonian University Medical College, Kraków, Poland; 3Department of Molecular Biology and Clinical Genetics, Faculty of Medicine, Jagiellonian University Medical College, Kraków, PolandCorrespondence: Natalia Celejewska-Wójcik 2nd Department of Internal Medicine, Faculty of Medicine, Jagiellonian University Medical College, ul. Jakubowskiego 2, Krakow, 30-688, PolandTel +48 12 400 30 53Fax +48 12 400 30 67Email natalia.celejewska@gmail.comPurpose: Lipid mediators, particularly eicosanoids, are associated with airway inflammation, especially with the eosinophilic influx. This study aimed to measure lipid mediators and cells in induced sputum, that could possibly reflect the inflammatory process in the bronchial tree of COPD subjects.Patients and Methods: Eighty patients diagnosed with COPD and 37 healthy controls participated in the study. Induced sputum samples were ascertained for differential cell count and induced sputum supernatant concentrations of selected eicosanoids by the means of gas chromatography/mass spectrometry and high-performance liquid chromatography/tandem mass spectrometry.Results: Increased sputum eosinophilia was associated with higher concentrations of selected proinflammatory eicosanoids. In COPD subjects prostaglandin D2 and 11-dehydro-thromboxane B2 correlated negatively with airway obstruction measured by FEV1 and FEV1/FVC values. COPD subjects with disease exacerbations during past 12 months had significantly higher concentrations of prostaglandin D2, 12-oxo-eicosatetraenoic acid and 5-oxo-eicosatetraenoic acid.Conclusion: Stable COPD is often associated with eosinophil influx in the lower airways and elevated concentrations of eicosanoids that is reflected by some disease characteristics.Keywords: chronic obstructive pulmonary disease, eosinophils, eicosanoids

Background To date, there has been no reliable in vitro test to either diagnose or differentiate nonsteroidal anti‐inflammatory drug (NSAID)–exacerbated respiratory disease (N‐ERD). The aim of the present study was to develop and validate an artificial neural network (ANN) for the prediction of N‐ERD in patients with asthma. Methods This study used a prospective database of patients with N‐ERD (n = 121) and aspirin‐tolerant (n = 82) who underwent aspirin challenge from May 2014 to May 2018. Eighteen parameters, including clinical characteristics, inflammatory phenotypes based on sputum cells, as well as eicosanoid levels in induced sputum supernatant (ISS) and urine were extracted for the ANN. Results The validation sensitivity of ANN was 94.12% (80.32%‐99.28%), specificity was 73.08% (52.21%‐88.43%), and accuracy was 85.00% (77.43%‐92.90%) for the prediction of N‐ERD. The area under the receiver operating curve was 0.83 (0.71‐0.90). Conclusions The designed ANN model seems to have powerful prediction capabilities to provide diagnosis of N‐ERD. Although it cannot replace the gold‐standard aspirin challenge test, the implementation of the ANN might provide an added value for identification of patients with N‐ERD. External validation in a large cohort is needed to confirm our results., Artificial neural network based on a set of clinical and biochemical parameters is able to stratify N‐ERD patients among asthmatic population. The sensitivity of the present artificial neural network is 94.12% and specificity is 73.08% for the prediction of N‐ERD. Artificial neural network could become a screening tool and allow to quickly identify patients with a high suspicion of N‐ERD

Aspirin-exacerbated respiratory disease (AERD) is characterized by overproduction of the pro-inflammatory eicosanoids. Although immunoglobulin E-mediated sensitization to aeroallergens is common among AERD patients, it does not belong to the defining disease characteristics. In this study of 133 AERD patients, we sought to find a relationship between sensitization to aeroallergens and local (leukotriene E4, prostaglandin E2 and prostaglandin D2) and/or systemic (leukotriene E4) production of arachidonic acid metabolites. Interestingly, a negative association between pro-inflammatory eicosanoid levels in induced sputum supernatant or urine and sensitization to aeroallergens was observed. This inverse relationship might suggest the presence of a protective effect of atopic sensitization to aeroallergens against stronger local airway inflammation and higher systemic AERD-related inflammatory activity.

Chronic exposure to PM

Background Aspirin desensitization followed by daily aspirin use is an effective treatment for aspirin-exacerbated respiratory disease (AERD). Objective To assess clinical features as well as genetic, immune, cytological, and biochemical biomarkers that might predict a positive response to high-dose aspirin therapy in AERD. Methods We enrolled 34 AERD patients with severe asthma who underwent aspirin desensitization followed by 52-week aspirin treatment (650 mg/d). At baseline and at 52 weeks, clinical assessment was performed; phenotypes based on induced sputum cells were identified; eicosanoid, cytokine, and chemokine levels in induced sputum supernatant were determined; and induced sputum expression of 94 genes was assessed. Responders to high-dose aspirin were defined as patients with improvement in 5-item Asthma Control Questionnaire score, 22-item Sino-Nasal Outcome Test (SNOT-22) score, and forced expiratory volume in 1 second at 52 weeks. Results There were 28 responders (82%). Positive baseline predictors of response included female sex (P=0.002), higher SNOT-22 score (P=0.03), higher blood eosinophil count (P=0.01), lower neutrophil percentage in induced sputum (P=0.003), higher expression of the hydroxyprostaglandin dehydrogenase gene, HPGD (P=0.004), and lower expression of the proteoglycan 2 gene, PRG2 (P=0.01). The best prediction model included Asthma Control Test and SNOT-22 scores, blood eosinophils, and total serum immunoglobulin E. Responders showed a marked decrease in sputum eosinophils but no changes in eicosanoid levels. Conclusions and clinical relevance Female sex, high blood eosinophil count, low sputum neutrophil percentage, severe nasal symptoms, high HPGD expression, and low PRG2 expression may predict a positive response to long-term high-dose aspirin therapy in patients with AERD.

Background Induced sputum (IS) allows to measure mediators of asthmatic inflammation in bronchial secretions. NSAID‐exacerbated respiratory disease (NERD) is recognized as a distinct asthma phenotype, usually with a severe course, eosinophilic airway inflammation, and increased production of pro‐inflammatory eicosanoids. A more insightful analysis of NERD patients has shown this phenotype to be nonhomogeneous. Objective We aimed to identify possible subphenotypes in a cohort of NERD patients with the means of latent class analysis (LCA). Methods A total of 95 asthma patients with aspirin hypersensitivity underwent sputum induction. High‐performance liquid chromatography or gas chromatography coupled with mass spectrometry was used to profile eicosanoids in induced sputum supernatant (ISS). Sixteen variables covering clinical characteristics, IS inflammatory cells, and eicosanoids were considered in the LCA. Results Three classes (subphenotypes) were distinguished within the NERD cohort. Class 1 subjects had mild‐to‐moderate asthma, an almost equal distribution of inflammatory cell patterns, the lowest concentrations of eicosanoids, and logLTE4/logPGE2 ratio. Class 2 represented severe asthma with impaired lung function despite high doses of steroids. High sputum eosinophilia was in line with higher pro‐inflammatory LTE4 in ISS and the highest logLTE4/logPGE2 ratio. Class 3 subjects had mild‐to‐moderate asthma and were also characterized by eosinophilic airway inflammation, yet increased production of pro‐ (LTE4, PGD2 and 11‐dehydro‐TBX2) was balanced by anti‐inflammatory PGE2. The value of logLTE4/logPGE2 was between values calculated for classes 1 and 3, similarly to disease control and severity. Conclusions LCA revealed three distinct NERD subphenotypes. Our results support a more complex pathobiology of aspirin hypersensitivity. Considering NERD heterogeneity, the relationship between inflammatory pathways and clinical manifestations of asthma may lead to more individualized treatment in difficult to treat patients in the future., Heterogeneity of NERD phenotype reflects differences in inflammatory response measured by airway cells and eicosanoids. Identifying subphenotypes provides a more insightful perception and suggests a need for more individualized approach. In aspect of logLTE4/logPGE2 ratio, latent class analysis assigned subject to different groups better than identification by disease severity or control emphasizing the heterogeneity in NERD subgroup. Abbreviations: LTE4, Leukotriene E4; NERD, NSAID‐exacerbated respiratory disease; PGE2, Prostaglandin E2

Background: Recently, we have reported that asthma is characterized by prothrombotic blood alterations, which were related to the low-grade inflammatory state. Inflammation, however, may also lead to vascular dysfunction. The aim of this study was to evaluate plasma levels of cellular fibronectin (cFN), a marker of vascular injury in asthmatics, and to analyze their impact on described previously prothrombotic blood alterations. Methods: In a cross-sectional study, we investigated 164 adult stable asthmatics and 72 matched controls. Plasma cFN was measured using an ELISA. Its relations to inflammation, thrombin generation, fibrinolytic capacity, expressed as clot lysis time (CLT), and platelet markers were evaluated. Results: Asthma was associated with 50.1% higher plasma cFN levels as compared with controls (p < 0.001, after adjustment for potential confounders). There was a positive association of cFN with asthma severity and inverse with the FEV1/VC index (beta=0.2 [95% CI: 0.13-0.28] and beta=-0.15 [95% CI: -0.23 to -0.07], respectively). In asthmatics cFN positively correlated with high-sensitivity C-reactive protein (beta= 0.24 [95% CI: 0.16-0.32]), fibrinogen (beta= 0.13 [95% CI: 0.04-0.21]), interleukin-6 (beta= 0.23 [95% CI: 0.15-0.3]), platelet factor 4 (beta= 0.14 [95% CI: 0.06-0.21]), plasminogen (beta= 0.11 [95% CI: 0.04-0.19]) and CLT (beta= 0.35 [95% CI: 0.28-0.42]). In both groups cFN was related to the endogenous thrombin potential (ETP) (beta= 0.51 [95% CI: 0.44-0.57], and beta= 0.17 [95% CI: 0.07-0.27], respectively). Multiple regression models showed that cFN was the most potent independent predictor of both ETP and CLT in asthmatics. Conclusion: Presented study is the first to show increased plasma cellular fibronectin in asthma, which is associated with disease severity, inflammation, and prothrombotic blood alterations. This novel observation suggests a previously unknown modulator of prothrombotic plasma properties in asthmatics.

Recently we have reported that asthma is associated with enhanced plasma thrombin formation, impaired fibrinolysis and platelet activation. In the present study we investigated whether described prothrombotic blood alterations might predispose to thromboembolic events or asthma exacerbations. In 164 adult asthmatics we assessed clinical events during 3-year follow-up and analyzed their associations with measured at baseline prothrombotic blood parameters. Data were obtained from 157 (95.7%) of the asthma patients. We documented 198 severe asthma exacerbations (64/year), which occurred in 53 subjects (34%). These patients were older (p = 0.004), had worse asthma control (p = 0.02) and lower spirometry values (p = 0.01), at baseline. Interestingly, this subgroup had longer clot lysis time (CLT), as well as lower α2-macroglobulin (p = 0.038 and p = 0.04, respectively, after adjustment for potential confounders). Increased CLT and lower α2-macroglobulin were demonstrated as independent predictors of asthma exacerbation in multiple regression model. Moreover, we documented two episodes of deep vein thrombosis (1.3%), and eight acute coronary syndromes (5.1%). Patients who experienced thromboembolic events (n = 10, 6.4%, 2.1%/year) had lower α2-macroglobulin (p = 0.04), without differences in efficiency of fibrinolysis and thrombin generation. Impaired fibrinolysis and lower levels of α2-macroglobulin might predispose to a higher rate of asthma exacerbations, suggesting new links between disturbed hemostasis and asthma.

Recently, we have reported that asthma is associated with enhanced plasma thrombin formation and impaired fibrinolysis. The mechanisms underlying the prothrombotic state in this disease are unknown. Our aim was to investigate whether prothrombotic alterations in asthmatics are associated with inflammation.We studied 164 adult, white, stable asthmatics and 72 controls matched for age, sex, body mass index (BMI), and smoking. Plasma tumor necrosis factor $\alpha$ ($TNF\alpha$), interleukin (IL)-6, and serum periostin were evaluated using ELISAs, and their associations with thrombin generation, fibrinolytic capacity, expressed as clot lysis time (CLT), and platelet markers were later analyzed. Asthma was characterized by 62% higher plasma IL-6 and 35% higher $TNF\alpha$ (both, p < 0.0001). Inflammatory cytokines were higher in sporadic and persistent asthmatics compared to controls, also after adjustment for potential confounders. IL-6 was inversely related to the forced expiratory volume in 1 s/vital capacity (FEV1/VC) spirometry index after correction for age, sex, and BMI. IL-6 and $TNF\alpha$ were associated with C-reactive protein in asthmatics ($\beta$ = 0.6 [95% CI, 0.54-0.67] and $\beta$ = 0.33 [95% CI, 0.25-0.41], respectively) and controls ($\beta$ = 0.43 [95% CI, 0.29-0.57] and $\beta$ = 0.33 [95% CI, 0.18-0.48], respectively). In asthma, IL-6 and $TNF\alpha$ positively correlated with the endogenous thrombin potential ($\beta$ = 0.35 [95% CI, 0.28-0.42] and \beta = 0.15 [95% CI, 0.07-0.23], respectively) but not with CLT or platelet markers. However, $TNF\alpha$ predicted CLT in a multiple linear regression model. Periostin was not associated with any hemostatic parameters. Enhanced thrombin generation is driven in asthma by a systemic inflammatory state mediated by IL-6 and to a lesser extent $TNF\alpha$, however, not periostin. $TNF\alpha$ might contribute to impaired fibrinolysis.

Background Aspirin desensitization (AD) is an effective and safe therapeutic option for patients with nonsteroidal anti-inflammatory drugs (NSAIDs)-exacerbated respiratory disease (N-ERD). The mechanisms driving its beneficial effects remain poorly understood. Objective To investigate the effect of long-term AD on clinical, biochemical and radiological changes in N-ERD patients. Methods The study group consisted of twenty-three individuals with N-ERD who underwent AD, followed by ingestion of 325 mg aspirin twice daily. Twenty patients completed the 52 weeks of AD. The following evaluations were conducted at baseline and in the 52nd week of AD: (i) clinical: asthma exacerbations, Asthma Control Test (ACT), Visual Analogue Scale (VAS) for the assessment of nasal symptoms; (ii) blood and induced sputum supernatant (ISS) periostin, (iii) phenotypes based on induced sputum (IS) cells, (iiii) ISS and nasal lavage (NL) concentration of prostaglandin D2 (PGD2), prostaglandin E2 (PGE2), tetranor-PGD-M, tetranor-PGE-M, 8-iso-PGE2, leukotriene B4 (LTB4), LTC4, LTD4 and LTE4, and urine LTE4. Results A significant improvement was observed in ACT (P = 0.02) and VAS score (P = 0.008) in the 52nd week of AD. ISS periostin and IS eosinophil count decreased significantly in the 52nd week of AD (P = 0.04 and P = 0.01, respectively). ISS and NL eicosanoid concentrations did not change following long-term AD. Conclusion and Clinical Relevance: AD is associated with a decrease in sputum periostin biosynthesis, which may prevent the recruitment of eosinophils into respiratory tissue and be one of explanation of the clinical benefits of AD. Long-term aspirin administration does not lead to an imbalance between pro- and anti-inflammatory ISS eicosanoids.

Background: Given increased risk of cardiovascular events in asthma we hypothesized that lipoprotein-associated phospholipase A2 (Lp-PLA2), an enzyme involved in atherosclerosis, is associated with proinflammatory and prothrombotic blood alterations in this disease. Methods: In 164 adult asthmatics (63 with severe asthma) we measured plasma Lp-PLA2 activity using the PLAC test. We determined its relations to inflammation and prothrombotic blood alterations. Results: In asthma, Lp-PLA2 was inversely related to the age (β = −0.1 [−0.18 to −0.02]) and was lower in women (n = 122 [74%], 205 [182–242] vs. 243 [203–262] nmol/min/ml, p = 0.001). Interestingly, Lp-PLA2 correlated negatively with the asthma severity score (β = −0.15 [−0.23 to −0.07]), being 10.3% higher in those with non-severe (mild or moderate) asthma (n = 101, 62%) as compared to the severe disease subtype (224 [191–261] vs. 203 [181–229], p = 0.006 after adjustment for potential confounders). Lp-PLA2 activity was positively related to the levels of low-density lipoprotein (β = 0.1 [0.02–0.18]), triglycerides (β = 0.11 [0.03–0.19]) and glucose (β = 0.1 [0.02–0.18]) and inversely to the tumor necrosis factor α (β = −0.27 [−0.35 to −0.2]), high sensitivity C-reactive protein (β = −0.1 [−0.19 to −0.02]) and fibrinogen (β = −0.12 [−0.21 to −0.03]), as well as prothrombin (β = −0.16 [−0.24 to −0.08]), and parameters describing thrombin generation potential, such as endogenous thrombin potential (β = −0.14 [−0.21 to −0.06]) and peak thrombin generated (β = −0.2 [−0.28 to −0.12]). Conclusions: Elevated Lp-PLA2 activity in non-severe asthmatics suggests increased atherosclerotic risk in this group. Lower Lp-PLA2 activity accompanied by its inverse relationship to inflammatory or prothrombotic blood biomarkers observed in turn in severe asthmatics might be related to the pathogenesis of more severe asthma phenotype. Keywords: Asthma, Cardiovascular risk, Coagulation, Lipoprotein-associated phospholipase A2, Inflammation

BACKGROUND A special regulatory role for prostaglandin E2 (PGE2 ) has been postulated in nonsteroidal anti-inflammatory drug (NSAID)-exacerbated respiratory disease (NERD). OBJECTIVE To investigate the effect of systemic aspirin (acetylsalicylic acid) administration on airway PGE2 biosynthesis in induced sputum supernatant (ISS) among subjects with NERD or aspirin-tolerant asthma with chronic rhinosinusitis with nasal polyposis (ATA-CRSwNP), as well as healthy controls (HC). METHODS Induced sputum (IS) was collected from patients with NERD (n = 26), ATA-CRSwNP (n = 17), and HC (n = 21) at baseline and after aspirin challenge. Sputum differential cell count and IS supernatant (ISS) levels of prostanoids, PGE2 , 8-iso-PGE2 , tetranor-PGE-M, 8-iso-PGF2 α, and leukotriene C4 , D4 , and E4 , were determined using mass spectrometry. Urinary excretion of LTE4 was measured by ELISA. RESULTS NERD subjects had elevated sputum eosinophilic count as compared to ATA-CRSwNP and HC (median NERD 9.1%, ATA-CRSwNP 2.1%, and HC 0.4%; P

Background There is evidence that altered blood coagulation and fibrinolysis are involved in the pathogenesis of asthma. Increased thromboembolic risk has been reported in asthmatics. Objective To investigate whether enhanced thrombin generation and impaired fibrinolysis occur in asthmatics. Methods Plasma thrombin generation profile together with a computational assessment of thrombin dynamics and fibrinolytic capacity expressed as clot lysis time (CLT) were determined in 164 consecutive patients with stable asthma and 72 controls matched for age, gender, weight and smoking. Results Asthma patients had 20.2% increased endogenous thrombin potential (ETP), 41.4% higher peak thrombin concentration, 61% higher maximal prothrombin conversion rate, 15.5% faster rate of thrombin formation (all, P < 0.0001) and 10% lower thrombin decay capacity (P = 0.0004) compared with controls. Asthmatics had also 14.4% longer CLT (P = 0.001) associated with 21.3% higher plasminogen activator inhibitor‐1 (PAI‐1) (P < 0.0001), and 13% higher plasma α2‐macroglobulin (P = 0.0002). Using ETP and CLT above 75th percentile of the control values as the cut‐off levels, we found increased risks of enhanced thrombin generation and hypofibrinolysis in asthmatics, also after correction for potential confounders. ETP and CLT were associated inversely with forced expiratory volume in 1 s/vital capacity (FEV1/VC) index, after adjustment for age and body mass index. Non‐allergic asthma (n = 70, 42.6%) was characterized by 17.5% longer CLT (P = 0.02), which positively associated with PAI‐1. Thrombin generation profile was not affected by allergy. Conclusion and Clinical Relevance Asthma is associated with enhanced thrombin generation and impaired fibrinolysis, which might contribute to thromboembolic events in this disease.

Objective/Background The diagnostics of latent tuberculosis infection in Poland using the tuberculin skin test is challenging due to the obligatory Bacillus Calmette–Guerin vaccinations. Interferon-gamma release assays are still very rarely used for diagnostics. We compared the tuberculin skin test and the QuantiFERON-TB Gold In-Tube test to evaluate the degree of latent tuberculosis infection in at-risk groups for tuberculosis (homeless, close contacts, periodic contacts, nursing-home attendees) and in healthy individuals. Methods QuantiFERON-TB Gold In-Tube tests were carried out on 785 individuals from the homeless ( n = 150), close contacts ( n = 171), periodic contacts ( n = 163), nursing-home attendees ( n = 152), and healthy individuals ( n = 149). The tuberculin skin test was performed on 129, 156, 147, 148, and 121 participants, respectively. We evaluated the (a) correlation between serum concentrations of interferon gamma and the tuberculin-skin-test induration diameter; (b) between the number of QuantiFERON-TB Gold In-Tube-positive results and the tuberculin-skin-test diameter in the studied groups; and (c) agreement between both tests and the kappa coefficient using the tuberculin-skin-test diameters of 5, 10, and 15 mm. Results Larger tuberculin-skin-test induration diameters were associated with elevated serum concentrations of interferon gamma. We found a positive correlation between the number of positive QuantiFERON-TB Gold In-Tube screening results and the tuberculin-skin-test induration diameter. The agreement between QuantiFERON-TB Gold In-Tube and tuberculin-skin-test screening results improved with increasing tuberculin-skin-test induration diameter. Conclusion Based on measures of tuberculin-skin-test induration diameter alone, it is difficult to diagnose latent tuberculosis infection with certainty. The agreement of the QuantiFERON-TB Gold In-Tube test increases with the tuberculin-skin-test diameter. Tuberculin-skin-test diameters larger than 15 mm are more likely to be associated with active infection.

Background: Eosinophilic airway inflammation has been demonstrated in 20%–40% of induced sputum samples from patients with stable COPD. Lipid mediators, like eicosanoids are associated with airway inflammation especially with the eosinophilic pattern. Objectives: To investigate the inflammatory cell patterns and eicosanoids in induced sputum in stable COPD. Methods: Induced sputum was collected in COPD and healthy control subjects. Sputum differential cell count and sputum supernatant concentrations of: 5-HETE, 12-HETE, 15-HETE, 5-oxo-ETE, 12-oxo-ETE, 15-oxo-ETE, LTD4, LTE4, LTB4, PGE2, 8-izo-PGE2, PGD2, PGA2, tetranor-PGE-M, tetranor-PGD-M, PGF2a, 8-izo-PGF2a, TBX2, 11-dehydro-TXB2 were measured. Results: In COPD subjects increased eosinophilia in induced sputum (>2%) was found in 42% of subjects (including 34% eosinophilic and 8% mixed eosinophilic and neutrophilic patterns). There was a statistically significant correlation between sputum eosinophilia and induced sputum concentrations of LTE4, LTD4, PGE2, 8-izo-PGE2, 15-HETE,15-oxo-ETE, PGD2, TBX2 and 11-dehydro-TBX2. Induced sputum concentrations of LTE4, LTD4, PGD2, PGE2, 8-izo-PGE2 were significantly higher in COPD subjects compared to healthy controls, whereas concentrations of tetranor-PGE-M and tetranor-PGD-M were lower. Frequent exacerbator COPD subjects had lower sputum but not blood eosinophilia. Conclusions: Stable COPD is often associated with eosinophil influx in the lower airways and elevated concentrations of eicosanoids. Increased sputum eosinophilia is not associated with frequent exacerbations of COPD.

Hypersensitivity to non-steroidal anti-inflammatory drugs(NSAID) is a distinct phenomenon of severe asthma. Eosinophils play a key role in NSAID hypersensitivity as a main source of prostanoides. The aim of the analysis is to assess the effect of omalizumab(OMA) on blood eosinophils count(Eo) in asthmatic patients with NSAID hypersensitivity compare to those who tolerate NSAID(non-NSAID). Out of 38 patients who started OMA therapy the positive history of NSAID hypersensitivity was reported by 14 patients, 24 tolerated NSAID. We evaluated Eo level and OMA clinical effectiveness in 16thweek and 52ndweek of therapy (P-value The baseline characteristic(demographic data, asthma severity) did not differ between study groups. The mean Eo did not differ between study groups at the baseline (mean±SD NSAID vs non-NSAID respectively: 578±681/μl vs 348±301/μl), although patients hypersensitive to NSAID had Eo≥500/μl almost 2 times more frequent. At 16thand 52ndweek the mean Eo did not change significantly. 4 patients(NSAID/non-NSAID=2/2) stopped the therapy due to subjective lack of benefit. In both groups, we observed significant improvement in ACQ and AQLQ scale, the reduction of annual exacerbations rate and the daily OCS dose in 16th and 52nd week. The improvement in asthma control parameters in 16thand 52ndweek did not differ between study groups. OMA therapy did not affect Eo blood in patients with NSAID-intolerance, regardless of good clinical response. OMA seems to be effective in patients suffering from severe allergic asthma independently of NSAID hypersensitivity status.(2015/19/B/NZ5/00096)

Aspirin desensitization is considered to be an effective and well-tolerated therapy for patients with Non-steroidal anti-inflammatory(NSAIDs)-Exacerbated Respiratory Disease (NERD). The aim of the present study was to investigate the influence of aspirin desensitization on inflammatory cell count in induced sputum and nasal lavage in fifteen NERD individuals subjected to one-year aspirin therapy. The decrease in induced sputum count of eosinophils and macrophages was observed. Clinical efficacy of aspirin therapy in improving nasal symptoms and quality of life in NERD patients was also confirmed.

A special regulatory role for prostaglandin ETo investigate the effect of systemic aspirin (acetylsalicylic acid) administration on airway PGEInduced sputum (IS) was collected from patients with NERD (n = 26), ATA-CRSwNP (n = 17), and HC (n = 21) at baseline and after aspirin challenge. Sputum differential cell count and IS supernatant (ISS) levels of prostanoids, PGENERD subjects had elevated sputum eosinophilic count as compared to ATA-CRSwNP and HC (median NERD 9.1%, ATA-CRSwNP 2.1%, and HC 0.4%; P 0.01). Baseline ISS levels of PGEAspirin-precipitated bronchoconstriction is associated with a decrease in airway PGE

Human rhinoviruses (RVs) are a major cause of exacerbations in asthma and other chronic airway diseases. A characteristic feature of asthmatic epithelium is goblet cell metaplasia and mucus hypersecretion. Bronchial epithelium is also an important source of lipid mediators, including pro- and antiinflammatory eicosanoids. By using air-liquid interface cultures of airway epithelium from patients with asthma and nonasthmatic control subjects, we compared RV16 replication-induced changes in mRNA expression of asthma candidate genes and eicosanoid production in the epithelium with or without IL-13-induced mucus metaplasia. Mucus metaplastic epithelium was characterized by a 20-fold less effective replication of RV16 and blunted changes in gene expression; this effect was seen to the same extent in patients with asthma and control subjects. We identified ciliary cells as the main target for RV16 by immunofluorescence imaging and demonstrated that the numbers of ciliary cells decreased in RV16-infected epithelium. RV16 infection of mucociliary epithelium resulted in overexpression of genes associated with bronchial remodeling (e.g., MUC5AC, FGF2, and HBEGF), induction of cyclooxygenase-2, and increased secretion of prostaglandins. These responses were similar in both studied groups. These data indicate that structural changes associated with mucus metaplasia renders airway epithelium less susceptible to RV infection. Thus, exacerbations of the lung disease caused by RV may result from severe impairment in mucociliary clearance or activation of immune defense rather than from preferential infection of mucus metaplastic epithelium. Repeated rhinoviral infections of compromised epithelium may contribute to the remodeling of the airways.

Introduction Evidence for heightened activation of blood coagulation in airways of asthmatic subjects is growing. Yet clinical relevance of altered hemostatic balance in circulating blood of asthmatics remains unclear. Increased thrombo-embolic risk has been reported in asthmatics. We hypothesized that asthma is associated with enhanced thrombin formation and impaired fibrinolysis. Aims and objectives Assess thrombin generation and fibrinolysis in asthmatics. Methods We determined plasma thrombin generation profile with computational assessment of thrombin dynamics and fibrinolytic potential expressed as clot lysis time (CLT), in 164 consecutive patients with stable asthma and 72 controls matched for age, sex, weight and smoking. Results Asthmtics had 20.2% increased endogenous thrombin potential (ETP), 41.4% higher peak thrombin concentration, 61% higher maximal prothrombin conversion rate, 15.5% faster rate of thrombin formation, and 10% lower thrombin decay capacity (all, p Conclusions Asthma is associated with enhanced thrombin generation and impaired fibrinolysis, which might contribute to thromboembolic events in this disease. Funds received:K/ZDS/002936, K/ZDS/004487, DEC-2013/09/B/NZ5/00758.

The aim of this pilot study was to evaluate changes in the concentration of prostaglandin E2 (PGE2) in induced sputum supernatant in 3 groups: sub- jects with NSAID-exacerbated respira- tory disease (NERD), aspirin tolerant asthma (ATA) and healthy controls (HC), before and after oral aspirin chal- lenge test. The study was conducted in the years 2014-2015 at the Clinical Department of the Pulmonology Clinic at the University Hospital in Cracow. 43 patients were enrolled in the study (NERD - n = 15, ATA - n = 15 and HC - n = 13). All of them underwent a placebo-controlled oral aspirin challenge. Sputum was induced 24 hours before the challenge and immediately after the test. Induced sputum was processed in order to obtain cystospin slides to depict inflammatory cell patterns and supernatants, in which PGE2 was measured. The concentration of PGE2 was determined using mass spectrometry coupled with gas chromatography (gas chromatography/mass spectrometry - GC/MS). After aspirin challenge, the concentration of PGE2 in induced sputum supernatant decreased in both asthmatics hypersensitive to aspirin (p = 0.01) and those who tolerated aspirin well (p = 0.17). The change in the healthy control group was not statistically significant. These results support the cyclooxygenase theory of PGE2 inhibition by aspirin. However, the mechanism of bronchoconstriction after aspirin administration alone in patients with NSAID-exacerbated respiratory disease remains unclear.

Eosinophilic granulomatosis with polyangiitis (EGPA) is associated with an inflammation and the presence of antineutrophil cytoplasmic antibodies (ANCA). Thus, we investigated the impact of ANCAs and eosinophilic inflammation on neutrophil activation and extracellular traps (NETs) formation.We recruited 29 patients in the remission of EGPA (17 ANCA-negative and 12 ANCA-positive, including 7 p-ANCA-positive and 5 c-ANCA-positive patients). Healthy donors' neutrophils were stimulated with EGPA patients' serum. NETs formation was assessed by immunofluorescence and scanning electron microscopy.EGPA patients presented enhanced ability to generate NETs compared to healthy subjects (20.3±8.2% vs. 2.7±1.5%, p=0.0036). However, there were no differences in NETs formation between ANCA-positive and ANCA-negative patients (23±11.2% vs. 17±6.1%, p=0.15). There was also no correlation between NETs generation and the amount of circulating DNA in EGPA patients. Among ANCA-positive patients, p-ANCA-positives showed the highest percentage of NETs as compared to cANCA-positive and ANCA-negative patients (27.3±10.3% vs. 17.8±10.5% and vs. 17±6.1%, both p0.01, respectively). Eosinophils number correlated with the percentage of NETs in the whole EGPA group (r=0.53, p=0.039), but we failed to observe the correlation with an eosinophil cationic protein (r=0.49, p=0.058).EGPA patients' serum has the ability to induce NETosis with no regard to the ANCA status in contrast to other vasculitides, where p-ANCA were considered as the main factor. Interestingly, NETs formation in EGPA patients connected with the number of eosinophils might be of major relevance. Further studies are required to assess which eosinophil-derived factors might be responsible for the neutrophils activation in EGPA patients.

The article below shows different forms, patomechanisms and diagnostics criteria of hypersensitivity to NSAIDs based on available literature as well as up to date outlook on implementing low salicylate diet as a treatment.

Background. Special regulatory role of eicosanoids has been postulated in aspirin-induced asthma. Objective. To investigate effects of aspirin on exhaled breath condensate (EBC) levels of eicosanoids in patients with asthma. Methods. We determined EBC eicosanoid concentrations using gas chromatography/mass spectrometry (GC-MS) and high-performance liquid chromatography/mass spectrometry (HPLC-MS2) or both. Determinations were performed at baseline and following bronchial aspirin challenge, in two well-defined phenotypes of asthma: aspirin-sensitive and aspirin-tolerant patients. Results. Aspirin precipitated bronchial reactions in all aspirin-sensitive, but in none of aspirin-tolerant patients (ATAs). At baseline, eicosanoids profile did not differ between both asthma groups except for lipoxygenation products: 5- and 15-hydroxyeicosatetraenoic acid (5-, 15-HETE) which were higher in aspirin-induced asthma (AIA) than inaspirin-tolerant subjects. Following aspirin challenge the total levels of cysteinyl-leukotrienes (cys-LTs) remained unchanged in both groups. The dose of aspirin had an effect on magnitude of the response of the exhaled cys-LTs and prostanoids levels only in AIA subjects. Conclusion. The high baseline eicosanoid profiling of lipoxygenation products 5- and 15-HETE in EBC makes it possible to detect alterations in aspirin-sensitive asthma. Cysteinyl-leukotrienes, and eoxins levels in EBC after bronchial aspirin administration in stable asthma patients cannot be used as a reliable diagnostic index for aspirin hypersensitivity.

Although peripheral nervous system involvement in patients with Churg-Strauss syndrome (CSS) has been described, little is known about its autonomic part. Autonomic nervous system (ANS) function can be assessed by studying heart rate variability (HRV) and a decrease in the spectrum of HRV correlates with ANS impairment.Out of 24 CSS patients we chose 12 (four males, eight females, aged 40 ± 8.3 years) in disease remission and without cardiac involvement. Twelve age- and sex-matched healthy volunteers served as a control group. All underwent 24-h electrocardiogram (ECG) Holter recordings. HRV was calculated from 1-h segments, including: total power (TP), ultra-low frequency (ULF), very low frequency (VLF), low frequency (LF), and high frequency (HF) powers as well as normalized LF (LF%) and HF (HF%) powers and the LF to HF power ratio (LF/HF).The CSS patients showed decreased HRV parameters in the 1-h domains: TP (2038 vs. 3622 ms(2), p = 0.001), HF (561 vs. 1574 ms(2), p0.001), LF (672 vs. 1050 ms(2), p0.01), and VLF (544 vs. 738 ms(2), p = 0.016). However, LF% and LF/HF ratio were markedly higher in CSS patients than in controls (53.4% vs. 39%, p0.001 and 1.1 vs. 0.64, p0.001), whereas HF% was lower in CSS than in controls (46.6% vs. 61%, p0.001). These results were independent of duration of the disease, eosinophil count, corticosteroids, or peripheral nerve involvement in the past.The CSS patients show impaired HRV parameters, indicating parasympathetic ANS dysfunction in addition to peripheral nervous system involvement.

Asthma is one of the key features of Churg-Strauss syndrome (CSS); however its course in the disease is unclear. The aim of this study was to analyze the asthma course in CSS patients.We conducted a retrospective study of 22 CSS patients. Medical documentation was studied and patients were questioned about asthma symptoms during follow-up visits, which took place at four points of the disease. These points, at which lung function tests were performed, were CSS diagnosis; introduction of treatment at hospital discharge; and assessment of the first and last clinical and laboratory CSS remissions. Asthma control and severity were assessed in compliance with current guidelines.In the asthmatic group examined, 21 patients had an adult-onset asthma, with a mean age of onset 35.5 ± 11.8 years, preceded by sinusitis (100%) and nasal polyposis (36%). Asthma at its onset was severe in 68.5%, moderate in 27%, and very poorly controlled in 21 patients. Atopy was present in 64% of patients. Onset of vasculitic symptoms and CSS diagnosis was accompanied by high blood eosinophilia (maximal 7.2 x 10(3)/l [1.2 - 32], asthma exacerbation with airway obstruction in 16 patients (mean values of FEV1 68.8 ± 17.5% and FVC 84.8 ± 19.6%), and lung involvement. After introducing the treatment and achieving stable remission, asthma severity/control and lung function tests (forced expiratory volume in 1 s 92.7 ± 13.3%, and forced vital capacity 101 ± 15.5%; p.001) improved.Asthma in CSS, although severe as its onset improves after achieving CSS remission.

Ostatnio opublikowane międzynarodowe raporty: Światowej Inicjatywy Zwalczania Astmy, PRACTicing ALLergology i Allergic Rhinitis and its Impact on Asthma rekomendują leki przeciwleukotrienowe w terapii astmy u dorosłych i u dzieci oraz w alergicznym nieżycie nosa. W niniejszym artykule przedstawiono argumenty za i przeciw lekom antyleukotrienowym w leczeniu obturacji dróg oddechowych.

Neurofibromatosis type 1 (NF1), referred to as von Recklinghausen’s disease, is a genetic disorder triggered by mutation of the NF1 gene, resulting in a lack of neurofibromin, which leads to abnormalities found in the peripheral nervous system and central nervous system, as well as in other organs. The disease is diagnosed early, usually in childhood by pediatricians. However, in some cases, the disease is clinically silent and remains undiagnosed or is recognized in the late adulthood. We report a case study of a 32-year-old female, who was referred to the pulmunologist with a suspicion of a lung tumor. The patient was admitted to the Pulmonology Department to investigate further the subpleural mass localized in the left lung found by chance in a chest X-ray. Physical examination revealed café-au-lait spots on her skin, several subcutaneous nodules which were confirmed by a histopathology to be consistent with neurofibroma. Further diagnostic testing, such as chest CT, PET and ophthalmological examination, led to diagnosis of neurofibromatosis type 1 with pulmonary involvement.

Rhabdomyoma is a rare tumor in the posterior mediastinum. We present a case of adult rhabdomyoma that occurred multifocally in the head, neck and mediastinum. There have been as few as four previous reports on rhabdomyoma in the posterior mediastinum. An 80-year-old white man was admitted to the Division of Pulmonary Diseases because of dyspnea, hemoptysis and fatigue. Physical examination revealed a tumor in the left submandibular area. Computed tomography (CT) of the head and neck showed a mass which caused a marked prominence of the left posterolateral wall of the pharynx. Chest CT revealed a contrast-enhancing tumor. The mass was 20 x 19 x 55 mm in size and was in contact with the lateral wall of the esophagus. Histological examination showed that the tumor was composed of cells typical of adult rhabdomyoma. The patient has been followed in our outpatient clinic for more than the last 7 years. To our knowledge, this is the fifth case of adult mediastinal rhabdomyoma. We believe that rhabdomyoma is a rare tumor which should be considered in a differential diagnosis of tumors detected in the posterior mediastinum.

Objectives. Churg–Strauss syndrome (CSS) is a rare necrotizing vasculitis associated with asthma, blood and tissue eosinophilia and granuloma formation. We wondered whether eosinophil accumulation in CSS results from the defect of intrinsic apoptosis pathway in blood eosinophils, leading to their prolonged survival. Methods. We analysed immunophenotype (flow cytometry), expression of apoptosis-related genes (real-time PCR) and spontaneous apoptosis in blood eosinophils isolated from nine patients in exacerbation (active CSS), seven patients in remission (inactive CSS) and 14 matched healthy subjects. Serum IL-5 levels were also measured. Results. In active CSS, blood eosinophils were characterized by small (

Background: A special regulatory role for prostaglandin E 2 has been postulated in aspirin-induced asthma. A study was undertaken to investigate the effects of aspirin on the systemic production of prostaglandin E 2 and cysteinyl leucotrienes in patients with asthma. Methods: The urinary concentrations were determined of two main prostaglandin E 2 metabolites (13,14-dihydro-15keto-PGE 2 using a commercial enzyme immunoassay and 9,15-dioxo-11α-hydroxy-2,3,4,5-tetranor-prostane-1,20-dioic acid by gas chromatography/mass spectrometry) and leucotriene E 4 using an immunoassay. Determinations were performed at baseline and following oral aspirin and celecoxib challenges in two well-defined asthma phenotypes: aspirin-sensitive and aspirin-tolerant patients. Results: Aspirin precipitated bronchial reactions in all aspirin-sensitive patients but in none of the aspirin-tolerant patients. Celecoxib 400 mg was well tolerated by all patients except for one with aspirin-induced asthma. At baseline, the mean levels of prostaglandin E 2 metabolites did not differ between the groups. Following different aspirin provocation doses, the mean levels of the two main prostaglandin E 2 metabolites were decreased in the aspirin-tolerant group but remained unchanged in the aspirin-sensitive group. The dose of aspirin had no effect on the magnitude of the response on the prostaglandin E 2 metabolites and its duration. In both groups, urinary prostaglandin E 2 metabolites decreased following celecoxib challenge. No correlation was found between prostaglandin E 2 metabolites and leucotriene E 4 . Conclusions: Aspirin-precipitated asthmatic attacks are not associated with changes in the systemic production of prostaglandin E 2 . In contrast, the systemic production of prostaglandin E 2 becomes depressed by aspirin in non-sensitive patients. This different response might indicate COX-1-dependent prostaglandin E 2 control of inflammatory cells in aspirin-induced asthma. Thus, PGE 2 is released during the clinical reactions to aspirin through an alternative COX-2 pathway. The clinical implications of this finding are in line with current observations of good tolerance of the selective COX-2 inhibitors in aspirin-sensitive patients.

Aspirin and other nonsteroidal anti-inflammatory drugs (NSAIDs) are among the most common causes of adverse drug reactions. Majority of them are of the hypersensitivity type. The two frequent clinical presentations of aspirin hypersensitivity are: aspirin-induced bronchial asthma/rhinosinusitis (AIA/R) and aspirin-induced urticaria/angioedema (AIU). The decisive diagnosis is based on provocation tests with aspirin, as the in vitro test does not hold diagnostic value as yet. Detailed protocols of oral, bronchial and nasal aspirin provocation tests are presented. Indications, contraindications for the tests, the rules of drug withdrawal and equipment are reviewed. Patient supervision and interpretations of the tests are proposed.

Background: Altered eicosanoid metabolismis a characteristic finding of aspirin induced asthma (AIA). Oral aspirin challenge is confirmatory diagnostic test for this clinical condition. Induced sputum allows to measure mediators of asthmatic inflammation in bronchial secretions. Objectives: The influence of aspirin on sputum supernatant concentration of eicosanoids during the oral challenge test was studied, using chromatography-mass spectrometry measurements. Methods: Induced sputum was collected before and following oral aspirin challenge. Sputum differential cell count and sputum supernatant concentrations of prostaglandin E 2 (PGE 2 ) was measured. Results: Aspirin precipitated bronchoconstriction was reported in all AIA (n=15) subjects, but in none of HC (n=13) and ATA (n=15). AIA was characterized by the dominance of the eosinophilic airway inflammation, whereas the paucigranulocytic phenotype was most frequent in HC. At baseline, mean induced sputum supernatant level of PGE 2 did not differ between the groups. Following the challenge, PGE 2 decreased in asthmatics patients independent of aspirin hypersensitivity (AIA p=0.002; ATAp=0.02), but not in HC (p=0.07). Cumulative doses of aspirin or the duration of the challenge had no effect on the magnitude of the PGE 2 alterations. Conclusions: During the oral aspirin challenge, PGE 2 decreases significantly in asthmatics as compared to healthy ones. The results support the theory on the inhibition of PGE 2 biosynthesis as a trigger for bronchoconstriction mediated by cysteinyl leukotrienes in AIA.

Background Induced sputum (IS) allows to measure mediators of asthmatic inflammation in bronchial secretions. The specific role of induced sputum supernatant (ISS) endogenous bioactive lipid mediators in subtypes of asthma is not well understood. Objective To investigate the interactions between airway inflammation and clinical phenotypes of asthma, we integrated induced sputum supernatant (ISS) eicosanoids and quantitative assessment of infiltrating cells into new subtypes with the means of latent class analysis (LCA). Methods One hundred and thirty-nine asthmatics with and without aspirin hypersensitivity underwent sputum induction. High-performance liquid chromatography or gas chromatography coupled with mass spectrometry was used to profile eicosanoids. Nineteen variables covering clinical characteristics, IS inflammatory cells and eicosanoids were considered in the LCA. Results Four phenotypic asthma classes were distinguished. Class 1 with mild-to-moderate asthma, chronic rhinosinusitis (CRS), high PGA2 in ISS and almost equal distribution of inflammation cell patterns. Class 3 subjects also had mild-to-moderate asthma but without upper airway symptoms. Induced sputum was often paucigranulocytic with low levels of lipid mediators. Classes 2 and 4 represented severe asthma with CRS and impaired lung function despite high doses of steroids. High blood and sputum eosinophilia was in line with high cysteinyl leukotrienes and PGD2 in ISS only in class 2. Class 4 subjects tended to have increased sputum neutrophilia and PGE2 in ISS. Aspirin hypersensitivity was most frequent among class 2 subjects. Conclusions & clinical relevance The LCA revealed four distinct asthma classes differing in eicosanoid pathways.

The association of nasal/sinus and bronchial symptoms with aspirin hypersensitivity is currently named aspirin-exacerbated respiratory disease (AERD). The application of biostatical methodology allowed us to identify unique AERD subphenotypes that can occur in regular clinical practice. According to the cyclooxygenase hypothesis, aspirin blocks cyclooxygenase enzymes (stronger – COX-1, weaker – COX-2), resulting in a decrease of the product concentration, catalyzed by those enzymes. There is a shift of arachidonic acid transformation from the cyclooxygenase pathway to 5-lipooxygenase, stimulating the synthesis of cysteinyl-leukotrienes (cys-LTs). The concentration of cys-LTs significantly increases after an aspirin challenge test in aspirin-exacerbate respiratory disease. This could be due to the fact that the pathogenesis of AERD is based on the assumption that a deficiency of prostaglandin E 2 (PGE 2 ) is responsible for bronchodilatation and cys-LTs production inhibition, which has a role in aspirin-induced bronchoconstriction. The results, non-invasively obtained induced sputum eicosanoids, have been acknowledged that a selective inhibition of PGE 2 biosynthesis by aspirin shifts the 5-lipoxygenase pathway from cys-LTs.

Objectives Given reports on the increased prevalence of thromboembolic incidents in patients with eosinophilic granulomatosis with polyangiitis (EGPA; Churg-Strauss syndrome), we investigated whether fibrin clot properties are unfavorably altered in EGPA. Methods Ex vivo plasma fibrin clot characteristics, including clot permeability, turbidimetry and efficiency of fibrinolysis using two assays, were investigated in 34 consecutive patients with remission in EGPA according to the Birmingham Vasculitis Activity Score version 3 (23 female, 11 male), aged 48 (range, 21–80) years. The control group comprised 34 age- and sex- matched volunteers. Results Compared with controls, patients with EGPA were characterized by denser fiber clots (estimated pore size, Ks, 7.30±0.93 vs 10.14±1.07 10−9 cm2), faster fibrin polymerization (lag phase in a turbidimetric curve, 41.8±3.6 vs 47.4±2.9 s), thicker fibrin fibers (maximum absorbance, ΔAbs, 0.87±0.09 vs 0.72±0.07), higher maximum levels of D-dimer released from clots (DDmax 4.10±0.46 vs 3.54±0.35 mg/L), and prolonged clot lysis time (t50%; 9.50±1.45 vs 7.56±0.87 min); all p

Background Isoprostanes are bioactive compounds formed by non-enzymatic oxidation of polyunsaturated fatty acids, mostly arachidonic, and markers of free radical generation during inflammation. In aspirin exacerbated respiratory disease (AERD), asthmatic symptoms are precipitated by ingestion of non-steroid anti-inflammatory drugs capable for pharmacologic inhibition of cyclooxygenase-1 isoenzyme. We investigated whether aspirin-provoked bronchoconstriction is accompanied by changes of isoprostanes in exhaled breath condensate (EBC). Methods EBC was collected from 28 AERD subjects and 25 aspirin-tolerant asthmatics before and after inhalatory aspirin challenge. Concentrations of 8-iso-PGF 2α , 8-iso-PGE 2 , and prostaglandin E 2 were measured using gas chromatography/mass spectrometry. Leukotriene E 4 was measured by immunoassay in urine samples collected before and after the challenge. Results Before the challenge, exhaled 8-iso-PGF 2α , 8-iso-PGE 2 , and PGE 2 levels did not differ between the study groups. 8-iso-PGE 2 level increased in AERD group only ( p = 0.014) as a result of the aspirin challenge. Urinary LTE 4 was elevated in AERD, both in baseline and post-challenge samples. Post-challenge airways 8-iso-PGE 2 correlated positively with urinary LTE 4 level ( p = 0.046), whereas it correlated negatively with the provocative dose of aspirin ( p = 0.027). Conclusion A significant increase of exhaled 8-iso-PGE 2 after inhalatory challenge with aspirin was selective and not present for the other isoprostane measured. This is a novel finding in AERD, suggesting that inhibition of cyclooxygenase may elicit 8-iso-PGE 2 production in a specific mechanism, contributing to bronchoconstriction and systemic overproduction of cysteinyl leukotrienes.

Background: The high-affinity receptor for immunoglobulin-E (IgE) (FcɛRI) plays a major role in the pathogenesis of allergy, but there are only two published studies on its α subunit (FcɛRIα) genetic variability in allergic diseases. Aims of the study: Mutational screening in the region of the FcɛRIα gene promoter and the first exon with subsequent genetic variability assessment in allergic patients and a random population sample. Methods: Allergic subjects were individuals with asthma or urticaria. Age- and sex-matched controls were randomly selected from a large population sample. Mutational screening was performed using a single-stranded conformational polymorphism and subsequent sequencing. Detected polymorphisms were genotyped by restriction fragment length polymorphism. Total serum IgE was measured in allergic subjects and controls. Skin prick tests, blood eosinophil count and aspirin challenge test were performed only in the subjects. A subgroup of the subjects was further characterized by autologous serum skin test, histamine release test, Phadiatop and IgE antibodies against staphylococcal enterotoxins. Results: Two linked polymorphisms −344 C>T and −95 T>C were found within the FcɛRIα gene. The allele −344 T frequency was 0.45 vs 0.37 (P = 0.33), and the allele −95 C frequency was 0.26 in subjects vs 0.30 in controls (P = 0.62). Serum IgE was significantly higher in subjects homozygous for the −344T allele (TT genotype) than in those carrying the −344 C allele (CT or CC genotype; P = 0.003), but this association was not detectable in controls. Conclusions: Our findings of genotype-related differences in IgE levels in allergic patients suggest an impact of −344 C>T but not −95 T>C gene polymorphism of FcɛRIα on total levels of IgE. The genetic variability in FcɛRIα at the −344 nucleotide of its regulatory sequence, though not related to atopy, predicts higher levels of the immunoglobulin.

Summary Background We have reported that in patients with chronic idiopathic urticaria (CIU) who reacted adversely to aspirin, the frequency of the −444C allele of the leukotriene C4 synthase gene (LTC4S) was higher than in patients who tolerated aspirin well. Objectives To study the pattern of aspirin-induced urticaria (AIU) in two families, with special interest on the polymorphisms of LTC4S (AA, AC, CC) and the glutathione S-transferase M1 and P1 genes (GSTM1 and GSTP1). Methods Of 74 patients with CIU and a history of aspirin hypersensitivity studied by us, two patients (probands) gave a family history of aspirin intolerance. Oral challenge tests with aspirin were carried out in members of these families. Genomic DNA samples were obtained from peripheral blood to study the polymorphisms of LTC4S, GSTM1 and GSTP1. Results In family 1 the aspirin challenge test confirmed AIU in three of five (60%) individuals, but in family 2 only in two of seven (29%). In both families, the variant genotypes of LTC4S (AC or CC) were present in the parents, but only one of them had CIU. In family 1, with both parents healthy, the three children had AIU; in two it was associated with variant LTC4S genotype. In family 2, urticaria following aspirin ingestion was present only with variant LTC4S genotype. In patients of both families with positive aspirin challenge test, deletion of the GSTM1 gene was present. Conclusions AIU aggregates in families inheriting the LTC4S−444C allele. Segregation of aspirin sensitivity in these families does not follow a clear Mendelian pattern. A common deletion of GSTM1, one of several enzymes involved in conjugation of a wide range of electrophilic substances with glutathione, was present in all individuals ascertained to have AIU.