Wojciech Barg

Metabolomics provide new insights on lung cancer staging and discrimination from chronic obstructive pulmonary disease.

Chronic obstructive pulmonary disease (COPD) and lung cancer are widespread lung diseases. Cigarette smoking is a high risk factor for both the diseases. COPD may increase the risk of developing lung cancer. Thus, it is crucial to be able to distinguish between these two pathological states, especially considering the early stages of lung cancer. Novel diagnostic and monitoring tools are required to properly determine lung cancer progression because this information directly impacts the type of the treatment prescribed. In this study, serum samples collected from 22 COPD and 77 lung cancer (TNM stages I, II, III, and IV) patients were analyzed. Then, a collection of NMR metabolic fingerprints was modeled using discriminant orthogonal partial least squares regression (OPLS-DA) and further interpreted by univariate statistics. The constructed discriminant models helped to successfully distinguish between the metabolic fingerprints of COPD and lung cancer patients (AUC training=0.972, AUC test=0.993), COPD and early lung cancer patients (AUC training=1.000, AUC test=1.000), and COPD and advanced lung cancer patients (AUC training=0.983, AUC test=1.000). Decreased acetate, citrate, and methanol levels together with the increased N-acetylated glycoproteins, leucine, lysine, mannose, choline, and lipid (CH3-(CH2)n-) levels were observed in all lung cancer patients compared with the COPD group. The evaluation of lung cancer progression was also successful using OPLS-DA (AUC training=0.811, AUC test=0.904). Based on the results, the following metabolite biomarkers may prove useful in distinguishing lung cancer states: isoleucine, acetoacetate, and creatine as well as the two NMR signals of N-acetylated glycoproteins and glycerol.

The use of magnesium in bronchial asthma: a new approach to an old problem

Abstract.Magnesium deficiency is a common electrolyte disorder in patients with acute severe asthma, but intracellular magnesium content better reflects its homeostasis than does its serum concentration. Magnesium takes part in many metabolic processes in the organism, including energy metabolism, protein and nucleic acid synthesis, cell cycle, the binding of substances to the plasma membrane, and maintenance of cytoskeletal and mitochondrial integrity. It also modulates ion transport and influences intracellular calcium concentration. Maintenance of the cells’ transmembrane gradient depends on the presence of magnesium, and hypomagnesemia may result in an increase in neuromuscular cell excitability. Magnesium is a cation modulating the smooth muscle contractility of different tissues: hypomagnesemia causes their contraction and hypermagnesemia their relaxation. Suggestions of a positive influence of magnesium in the treatment of asthma exacerbation have been known for a long time, but research results differ. A single dose of intravenous magnesium sulfate given to patients with acute asthma exacerbation has been shown to be safe, but its efficiency is still under discussion. According to the Global Initiative for Asthma GINA-2005, magnesium sulfate administration is not recommended for routine treatment, but it is permitted in patients with severe asthma exacerbation not responding to treatment (evidence category A). Recommendations of the British Thoracic Society allow one dose of magnesium sulfate to patients with acute severe asthma exacerbation and inadequate initial response to broncho-dilating inhalation treatment (evidence category A). Future investigations should help to establish the indications for magnesium use in the treatment of acute asthma exacerbations as well as the magnesium dose and the scheme of its administration.

Latex Allergy and Sensitization in Children with Spina bifida

Background: The purpose of this study was to investigate the prevalence rate of latex sensitization and latex allergy among children with spina bifida and to evaluate risk factors for natural rubber latex hypersensitivity. Methods: A total of 34 children between 2.5 and 17 years of age participated in the study. Participants completed a questionnaire and underwent skin prick tests with latex, common aeroallergens and food allergens as well as measurements of specific IgE to latex and food allergens (RAST CAP). Results: The prevalence of latex sensitization and latex allergy was estimated to be 32.4 and 18.8%, respectively. The most common reported clinical manifestation of latex allergy was urticaria. Three out of six symptomatic patients reported anaphylactic reactions. Conclusion: We found that major risk factors for latex sensitization were atopy and a history of numerous operations.

The G/G genotype of transforming growth factor beta 1 (TGF-β1) single nucleotide (+915G/C) polymorphism coincident with other host and environmental factors is associated with irreversible bronchoconstriction in asthmatics

Irreversible airflow obstruction may develop in some cases of asthma even in absence of known risk factors such as smoking and environmental insults and despite implementing apparently appropriate therapy. This implies that genetic factors may significantly contribute to determining the severity in the course of the disease. The published reports on genetic predisposition to irreversible bronchoconstriction in asthma, however, are relatively scarce, and disregard its potential association with transforming growth factor (TGF)‐β1 gene polymorphism despite established role that TGF‐β1 plays in airway remodelling. We tested TGF‐β1 single‐nucleotide polymorphisms (SNPs) at position +869 of codon 10 (leucine or proline) and position +915 of codon 25 (arginine or proline) for association with irreversible bronchoconstriction in a case–control study involving 110 patients with asthma and 109 controls. Multivariate logistic regression analysis revealed that genotype G/G at codon 25 was significantly associated with irreversible bronchoconstriction in asthmatics (odds ratio = 4.44; 95% confidence interval: 1.00–19.61; P = 0.05), but only after adjustment for gender, disease duration and smoking index. The influence of SNPs at codon 10 on irreversible airway obstruction was not significant. Our results suggest that presence of SNP (+915G/G) at codon 25 in TGF‐β1 gene may predispose to the development of irreversible bronchoconstriction in asthmatic patients, but only when coincident with the male gender, habitual smoking and relevant duration of the disease.

Airway inflammation in patients with chronic non-asthmatic cough

Introduction Chronic non-asthmatic cough (CC) is a clinical challenge and underlying pathophysiological mechanisms remain still not completely understood. One of the most common comorbidities in CC is gastro-oesophageal reflux disease (GORD). Airway epithelium damage can contribute to airway inflammation in CC. Aims We studied airway inflammation in patients with CC compared to healthy controls. Patients with GORD were treated with proton pump inhibitors (PPI) and cough response to PPI was evaluated. Patients and methods Sputum was induced in 41 adults with CC and 20 healthy non-smokers who were age and sex matched. We compared sputum differential cell count by cytospin and cytokine and chemokine production at the mRNA and/or protein levels by real-time (RT)-PCR and cytokine bead array (CBA), between patients with CC and healthy subjects. Furthermore we studied airway inflammation in patients with different comorbidities. Results No differences in sputum differential cell counts were observed between patients with CC and healthy subjects. Sputum monocyte chemoattractant protein-1 (MCP-1) protein levels were significantly higher in patients when compared to controls. Thymic stromal lymphopoietin (TSLP) mRNA was significantly more often expressed in sputum of patients with CC than from healthy controls. Sputum transforming growth factor (TGF)-β levels did not differ between patients and controls, but were significantly lower in the PPI responders compared to the non-responders; p=0.047. There is no evidence for impaired T helper cell (Th)1/Th2/Th17 balance in CC. Patients with reflux oesophagitis (RO) have significantly more sputum eosinophils than patients without RO. Conclusions CC is a condition presenting with different disease phenotypes. High sputum MCP-1 levels are present in a large group of patients with CC and a majority of these patients with CC have increased sputum TSLP levels, most likely produced by damaged airway epithelial cells.

Food-dependent, exercise-induced anaphylaxis triggered by co-incidence of culprit food, physical effort and a very high dose of ibuprofen or menstruation: a case report

Address for correspondence: Krzysztof Gomułka MD, PhD, Department of Internal Diseases and Allergology, Medical University of Wroclaw, 66 Marii Curie-Skłodowskiej St, 50-369 Wroclaw, Poland, phone: +48 71 784 25 28, e-mail: kgomulka@wp.pl Received: 11.12.2015, accepted: 22.04.2016. Food-dependent, exercise-induced anaphylaxis triggered by co-incidence of culprit food, physical effort and a very high dose of ibuprofen or menstruation: a case report

CD164 as a Basophil Activation Marker.

Introduction of flow cytometric technique to the research on basophil activation has led to significant achievements in allergy diagnosis in vitro. Most of the studies employing the flow cytometry used CD63 as a marker of basophil activation and only some of them used CD203c. Recently discovered basophil activation markers, including CD164, opened new possibilities for solving majority of current diagnostic needs. Use of allergen-induced CD164 upregulation in diagnosis of pollen allergy has been validated, and this encourages to further studies on other diagnostic usefulness of this marker. There are some hopeful data indicating that it might be useful in diagnosis of allergy caused by variety of other allergens, including drug allergens. Although CD164 upregulation as a marker of basophil activation is a promising and powerful diagnostic tool, it still requires a lot of both basic research and comparative studies with older and well known markers, in order to select the best of them. A research on basophil CD164 upregulation caused by various stimuli offers a good possibility to increase our knowledge of basophils involvement in allergic inflammation. Moreover, this might trigger a variety of pharmacological studies with known and new anti-inflammatory drugs in the future.

The diagnostic usefulness of the basophil activation test (BAT)with annexin V in an allergy to Alternaria alternata

BACKGROUND The basophil activation test (BAT) is an effective diagnostic tool in mold allergy, which is still not sufficiently known. OBJECTIVES The aim of our study was to assess the degree of annexin V binding to the surface of the basophil cell membrane after stimulation with anti-immunoglobulin E (anti-IgE) and Alternaria alternata allergenic extract. MATERIAL AND METHODS Alternaria alternata allergic patients (n = 32) and healthy volunteers (n = 33) were evaluated using skin prick tests (SPT), quantification of specific IgE (sIgE) and the BAT. Basophil activation was detected as a percentage degree of annexin V binding to the surface of the basophil cell membrane. RESULTS Receiver operating characteristic (ROC) curve analysis yielded a threshold value of 4.95% of activated basophils when the tested group and control group were studied, with a sensitivity and specificity of 100% (area under curve [AUC] = 1; p = 0.00000) for 100 SBU/mL Alternaria alternata allergen extract. The threshold value was 10.28% with a sensitivity of 93.8% and specificity of 100% (AUC = 0.98958; p = 0.00000) for 10 SBU/mL mold extract, and 9.37% with a sensitivity of 90.3% and specificity of 100% (AUC = 0.96307; p = 0.00000) for 1 SBU/mL Alternaria alternata allergen extract. The method was least efficacious in antiIgE stimulation, where the threshold value was 5.48% with a sensitivity of 90.6% and specificity of 30.3% (AUC = 0.46780; p = 0.67039). CONCLUSIONS The BAT with annexin V and sIgE measurement against Alternaria alternata increase the capability of a diagnostic laboratory for detecting mold sensitization. Both methods may certainly replace SPT, which are currently routinely used in allergy diagnosis. Annexin V may be considered a new basophil activation marker with an efficacy comparable to that of CD63 or CD203c.

Metabolomics of chronic obstructive pulmonary disease and obstructive sleep apnea syndrome: response to Maniscalco and Motta.

We appreciate Maniscalco and Motta’s comments on our recently published article “Fusion of the 1H NMR data of serum, urine and exhaled breath condensate in order to discriminate chronic obstructive pulmonary disease and obstructive sleep apnea syndrome” (Zabek et al. 2015) and we are grateful for the opportunity to clarify a number of points from our work. We are glad that the authors appreciated our data analysis and interpretation. Fusion of metabolomic data is an outstanding tool for a comprehensive description of patient’s phenotype that combines metabolic profiles acquired in different domains. Such an integrative approach is highly valued in the so-called ‘breathomics’ (Smolinska et al. 2014) for a better understanding of the complex respiratory diseases and discovering their biomarkers (Sterk et al. 2013). Our work was an attempt to provide a novel comprehensive description of metabolic phenotype and to test its utility in order to discriminate between two closely related respiratory pathological conditions. We performed a very careful selection of patients and those with an overlap syndrome who suffered from both COPD and OSAS (a total of 21 individuals) were not considered in our study. A further reduction in the number of patients was obtained by eliminating patients who did not deliver samples of all three biofluids for analysis. Therefore, even though a total of 85 serum, 91 urine and 82 EBC samples were collected, only 46 individuals (18 patients with COPD and 28 patients with the OSA syndrome) were finally included in the study. The selection of samples is in agreement with the assumption that changes in the level of a metabolite or a combination of metabolites present in different biofluids may be characteristic for a disease at all stages of its progression, regardless of the degree of the severity of the disease, obesity or smoking habits of the patients. Therefore, in our opinion, the hypothesis suggested by Maniscalco and Motta that the ‘lower specificity’ of the models combining EBC and other biofluids might be related to the severity of the disease would require further verification using specifically designed data. In order to assess the effect of the severity of a disease, obesity or smoking habits on the metabolic profiles, analysis of data in which a strict categorization of COPD and OSAS patients according to their clinical profiles is required. Unfortunately, it was not possible to present an analysis of such designed data in our article due to the small number of samples. Stratification of patients based on spirometric results was out of the scope of this study. A comparison of our results and the results presented in the recent publication authored by Maniscalco and Motta (Paris et al. 2015) is hindered due to the differences in normalization methods that were used. We are currently developing a more comprehensive clinical trial that we hope will provide a clearer answer to the questions that have been raised. It is our conviction that our work will be a useful contribution to the field of metabolomics and will help to better understand the metabolic mechanisms of respiratory pathologies.