Genome-Wide DNA Methylation and Gene Expression in Patients with Indolent Systemic Mastocytosis.

Mastocytosis is a clinically heterogenous, usually acquired disease of the mast cells with a survival time that depends on the time of onset. It ranges from skin-limited to systemic disease, including indolent and more aggressive variants. The presence of the oncogenic KIT p. D816V gene somatic mutation is a crucial element in the pathogenesis. However, further epigenetic regulation may also affect the expression of genes that are relevant to the pathology. Epigenetic alterations are responsible for regulating the expression of genes that do not modify the DNA sequence. In general, it is accepted that DNA methylation inhibits the binding of transcription factors, thereby down-regulating gene expression. However, so far, little is known about the epigenetic factors leading to the clinical onset of mastocytosis. Therefore, it is essential to identify possible epigenetic predictors, indicators of disease progression, and their link to the clinical picture to establish appropriate management and a therapeutic strategy. The aim of this study was to analyze genome-wide methylation profiles to identify differentially methylated regions (DMRs) in patients with mastocytosis compared to healthy individuals, as well as the genes located in those regulatory regions. Genome-wide DNA methylation profiling was performed in peripheral blood collected from 80 adult patients with indolent systemic mastocytosis (ISM), the most prevalent subvariant of mastocytosis, and 40 healthy adult volunteers. A total of 117 DNA samples met the criteria for the bisulfide conversion step and microarray analysis. Genome-wide DNA methylation analysis was performed using a MethylationEPIC BeadChip kit. Further analysis was focused on the genomic regions rather than individual CpG sites. Co-methylated regions (CMRs) were assigned via the CoMeBack method. To identify DMRs between the groups, a linear regression model with age as the covariate on CMRs was performed using Limma. Using the available data for cases only, an association analysis was performed between methylation status and tryptase levels, as well as the context of allergy, and anaphylaxis. KEGG pathway mapping was used to identify genes differentially expressed in anaphylaxis. Based on the DNA methylation results, the expression of 18 genes was then analyzed via real-time PCR in 20 patients with mastocytosis and 20 healthy adults. A comparison of the genome-wide DNA methylation profile between the mastocytosis patients and healthy controls revealed significant differences in the methylation levels of 85 selected CMRs. Among those, the most intriguing CMRs are 31 genes located within the regulatory regions. In addition, among the 10 CMRs located in the promoter regions, 4 and 6 regions were found to be either hypo- or hypermethylated, respectively. Importantly, three oncogenes-FOXQ1, TWIST1, and ERG-were identified as differentially methylated in mastocytosis patients, for the first time. Functional annotation revealed the most important biological processes in which the differentially methylated genes were involved as transcription, multicellular development, and signal transduction. The biological process related to histone H2A monoubiquitination (GO:0035518) was found to be enriched in association with higher tryptase levels, which may be associated with more aberrant mast cells and, therefore, more atypical mast cell disease. The signal in the BAIAP2 gene was detected in the context of anaphylaxis, but no significant differential methylation was found in the context of allergy. Furthermore, increased expression of genes encoding integral membrane components (GRM2 and KRTCAP3) was found in mastocytosis patients. This study confirms that patients with mastocytosis differ significantly in terms of methylation levels in selected CMRs of genes involved in specific molecular processes. The results of gene expression profiling indicate the increased expression of genes belonging to the integral component of the membrane in mastocytosis patients (GRM2 and KRTCAP3). Further work is warranted, especially in relation to the disease subvariants, to identify links between the methylation status and the symptoms and novel therapeutic targets.

A Challenge for Allergologist: Application of Allergy Diagnostic Methods in Mast Cell Disorders.

Primary and secondary mast cell activation syndromes (MCAS) can occur in patients with mastocytosis. During the past few years our knowledge about the pathogenesis and disease-triggering mechanisms in MCAS and mastocytosis have increased substantially. Whereas mastocytosis is characterized by an accumulation of neoplastic (clonal) mast cells (MC) in various organ systems, MCAS is defined by a massive and systemic activation of these cells. Mast cells are crucial effector cells in allergic diseases, thus their elevated number and activation can cause severe anaphylactic reactions and MCAS in patients with mastocytosis. However, these cells may also degranulate spontaneously or degranulate in response to non-allergic triggers leading to clinical symptoms. In mastocytosis patients, such symptoms may lead to the diagnosis of a primary MCAS. The diagnosis of a concomitant allergy in mastocytosis patients is challenging. In these patients, a mixed form (primary and secondary) of MCAS may be diagnosed. These patients may also suffer from life-threatening anaphylactic reactions when exposed to allergens. In these cases, the possibility of severe side effects of in vivo provocations can sometimes also limit diagnostic evaluations. In the current article, we discuss the diagnosis and management of patients suffering from mastocytosis and concomitant MCAS, with special emphasis on novel diagnostic tests and management, including allergen microarrays, recombinant allergen analysis, basophil activation tests, optimal prophylaxis, and specific therapies.

Results from a Genome-Wide Association Study (GWAS) in Mastocytosis Reveal New Gene Polymorphisms Associated with WHO Subgroups.

Mastocytosis is rare disease in which genetic predisposition is not fully understood. The aim of this study was to analyze associations between mastocytosis and single nucleotide polymorphisms (SNPs) by a genome-wide association study (GWAS) approach. A total of 234 patients were enrolled in our study, including 141 with cutaneous mastocytosis (CM; 78 children and 63 adults) and 93 with systemic mastocytosis (SM, all adults). The control group consisted of 5606 healthy individuals. DNA samples from saliva or blood were genotyped for 551 945 variants using DNA microarrays. The prevalence of certain SNPs was found to vary substantially when comparing patients and healthy controls: rs10838094 of 5OR51Q1 was less frequently detected in CM and SM patients (OR = 0.2071, p = 2.21 × 10-29), rs80138802 in ABCA2 (OR = 5.739, p = 1.98 × 10-28), and rs11845537 in OTX2-AS1 (rs11845537, OR = 6.587, p = 6.16 × 10-17) were more frequently detected in CM in children and adults. Additionally, we found that rs2279343 in CYP2B6 and rs7601511 in RPTN are less prevalent in CM compared to controls. We identified a number of hitherto unknown associations between certain SNPs and CM and/or SM. Whether these associations are clinically relevant concerning diagnosis, prognosis, or prevention remains to be determined in future studies.

Factors influencing adherence to treatment in COPD patients and its relationship with disease exacerbations.

INTRODUCTION: Chronic obstructive pulmonary disease (COPD) is one of the leading causes of morbidity and mortality in the world. Systematic treatment of COPD decreases symptoms and reduces the frequency of exacerbations and hospitalisations because of the disease. It is estimated that only 50% of patients use prescribed drugs systematically. The aim of this study was to identify the factors which can influence adherence to treatment of the patients who were treated due to exacerbation of COPD. MATERIAL AND METHODS: A questionnaire probe was conducted on 49 patients hospitalised at the Regional Lung and Tuberculosis Hospital in Olsztyn, Poland due to COPD exacerbation. The assessed variables were: quality of life and adherence to treatment 30 days after discharge from hospital in relationship with demographic factors, social status, disease and hospitalisation course, and relief after systematic treatment. RESULTS: Most of the patients assessed their health condition as poor and the disease as limiting their everyday social and occupational activity. 30 days after discharge from hospital the adherence rate to therapy was only 67%. There was an association between systematic treatment and the rate of exacerbations (P = 0.045) and hospitalisations (P = 0.005) but also clinical benefit after long-term treatment (P = 0.023). There were no associations between adherence to treatment and sex, place of residence, education or occupation. CONCLUSIONS: Lack of systematic treatment is the main risk factor for COPD exacerbations and hospitalisation rate. A subjective sense of relief after drugs is a factor improving patients' compliance.

The influence of nutritional habits, body mass index and intestinal microbiota in mastocytosis on clinical symptoms using conventional culture and next generation sequencing.

BACKGROUND: Mastocytosis is a rare neoplastic disease of the bone marrow associated with the proliferation and accumulation of mast cells in various internal organs, including the gastrointestinal tract. There are few studies describing the gut microbiome of patients with mastocytosis using next generation sequencing supported using traditional culture methods. The aims of the study were, firstly, the determination of nutrition habits, composition of the intestinal microflora and BMI in mastocytosis, and secondly, analysis of mastocytosis severity and symptoms depending on the composition of the intestinal microflora. METHODS: The study included 47 patients with indolent systemic mastocytosis and 18 healthy controls. All participants gave their informed consent to participate in the study. The study consisted of 3 parts: I-clinical assessment, II - examination of the intestinal microflora using the biochemical method, III - 16S rRNA sequencing. RESULTS: The nutrition habits and BMI of mastocytosis patients were similar to controls; however, most patients with mastocytosis had a low dietary vitamin and mineral content. As many as 94.5% of patients had too little fiber intake and mineral content. The most common cause of the abnormal stool test result with traditional culture was a titer of E. coli <106 . The low richness of microbiota species indicated by the Simpson index was observed in mastocytosis, p = 0.04. There were no significant differences in the composition of the intestinal microflora depending on the type of mastocytosis; however, the tryptase level correlated with the amount of Suterella, Barnesiellaceae, Eubacterium, Odoribacter, and Anaerostipes. CONCLUSIONS: The nutritional habits and BMI of mastocytosis patients are similar to the general population, except for too little fiber intake and mineral content. The gastrointestinal symptoms of mastocytosis patients may be related to the low richness of microbiota species and the amount of Suterella, Barnesiellaceae, Eubacterium, Odoribacter, Anaerostipes, which correlated with tryptase levels.

Dietary Patterns of Treatment-Resistant Depression Patients.

Depression is a common mental disorder that occurs all over the world with treatment resistance commonly seen in clinical practice. Ketamine exhibits an antidepressant that is more often used in the case of treatment-resistant depression (TRD) in MDD and BP. Research emphasizes that a healthy diet and the nutrients it contains can lower the risk of developing depression and form a strategy that supports conventional treatment. The aim of the study was to evaluate the patients' diet and to analyze the effect of ketamine on food intake among patients with TRD. The study involved 15 patients suffering from treatment-resistant depression and 15 healthy volunteers. The data required for the analysis were collected using the food frequency questionnaire (FFQ) and 4-day food diaries. The study group was statistically significantly less likely to consume milk and plain milk beverages, plain white cheese, wholemeal bread, various vegetables, wine, and drinks. Our results show several disorders in the eating habits of patients with treatment-resistant depression. After the administration of ketamine, the patients consumed significantly less protein, fats, monounsaturated fatty acids (MUFA) and polyunsaturated fatty acids (PUFA), fiber, tryptophan, vitamins, and minerals compared to the control group. There is a lack of research describing the effects of ketamine on nutrition. In order to confirm the results of the study, more participants are required, and the assessment of food diaries filled in at the patient's home with a longer interval after the last dose of ketamine as well.

DNA methylation profile in patients with indolent systemic mastocytosis.

BACKGROUND: Mastocytosis is a clinically heterogeneous, usually acquired disease of the mast cells with a survival time that depends on the onset of the disease and ranges from skin-limited to systemic disease, including indolent and more aggressive variants. The crucial element in pathogenesis is the presence of oncogenic KIT somatic mutation D816V. Further epigenetic alterations are responsible for regulating the expression of genes. It is essential to identify indicators of disease progression, and the specific clinical picture to establish an appropriate therapeutic strategy. OBJECTIVE: The aim of this study was to analyze the relation of mastocytosis symptoms and epigenetic changes, and to identify epigenetic predictors of the disease. METHODS: Global DNA methylation profile analysis was performed in peripheral blood collected from 73 patients with indolent systemic mastocytosis (ISM) and 43 healthy adult volunteers. Levels of 5-methylcytosine (5-mC) and 5-hydroxymethylcytosine (5-hmC) were determined using an ELISA-based method, while the methylation of the Alu and LINE-1 repeats were assayed with the quantitative methylation-specific PCR technique. A questionnaire interview was conducted among the study participants to collect data on possible epigenetic modifiers. Additionally, the methylation profile was compared between three human mast cell lines: ROSA KIT D816V, ROSA KIT WT, and HMC-1.1 KIT V560G, in order to assess the association between KIT mutations and methylation profile. RESULTS: A significantly lower level of DNA hydroxymethylation (5-hmC) in the blood was found in patients with ISM as compared to the controls (0.022% vs. 0.042%, p = 0.0001). Differences in the markers of global DNA methylation (5-mC, Alu, LINE-1) were not statistically significant, although they did indicate generally higher DNA methylation in patients with mastocytosis. The 5-hmC level was significantly associated with allergy (p = 0.011) in patients with ISM, showing a higher level of 5-hmC in patients with allergy as compared to patients without allergy. The in vitro study revealed significant differences between the studied cell lines at the level of 5-mC, Alu, and LINE-1. CONCLUSIONS: This study confirms that epigenetic changes are involved in mastocytosis, and suggests that allergy may be an important epigenetic modifier of the disease. A possible association between KIT mutations and methylation status observed in human mast cell lines requires further investigation in human studies. CLINICAL IMPLICATIONS: Epigenetic alterations are involved in mastocytosis pathology. The possible role of allergy as an important epigenetic modifier suggests the more impaired function of mast cells in ISM patients without allergy. CAPSULE SUMMARY: Decreased DNA demethylation in the blood DNA of patients with ISM confirms that epigenetic alterations are involved in mastocytosis pathology. We observed a possible role of allergy as an important epigenetic modifier. There is a possible association between KIT mutations and the methylation status observed in human mast cell lines.

How to diagnose mast cell activation syndrome: practical considerations.

Mast cells (MCs) are an important component of the immune system. Their physiological function is involved in multiple areas of human physiology, thus symptoms of their increased activation vary greatly from severe allergic reactions, such as anaphylaxis, to chronic symptoms, such as depression or osteoporosis. Studies on mastocytosis revealed a subgroup of patients presenting symptoms of increased MC degranulation, defined as mast cell activation syndrome (MCAS). This population includes patients with primary MCAS with clonal abnormal MCs, who do not fulfill the criteria for mastocytosis. These symptoms often overlap with comorbidities, which makes the diagnosis and treatment of MCAS difficult. The syndrome is diagnosed on the basis of 3 criteria: 1) the presence of typical symptoms; 2) elevation of serum tryptase levels; and 3) response to anti-mediator treatment. The diagnosis of MCAS is important especially in patients with anaphylaxis or osteoporosis who require the use of an epinephrine emergency kit and insect venom immunotherapy. In this review, genetic mechanisms and typical symptoms of MCAS as well as its diagnostic criteria and implications were discussed, with a special emphasis on practical guidance with the aim to improve patient care.

Dietary Habits in Children with Respiratory Allergies: A Single-Center Polish Pilot Study.

BACKGROUND: The rising trend in allergic diseases has developed in parallel with the increasing prevalence of obesity, suggesting a possible association. The links between eating habits and allergies have not been sufficiently clarified. AIM: To evaluate the nutritional status, eating habits, and risk factors of obesity and pulmonary function in children with allergic rhinitis. MATERIALS AND METHODS: We evaluated 106 children with allergic rhinitis (mean age 12.1 ± 3.4 years; M/F 60/46) from the Department of Allergology. Clinical data were collected regarding allergies, physical activity, nutritional status (Bodystat), dietary habits (Food Frequency Questionnaire validated for the Polish population), skin prick test with aeroallergens (Allergopharma), and spirometry (Jaeger). RESULTS: All children suffered from allergic rhinitis; among them, 43 (40.6%) presented symptoms of asthma. There were differences between children with only allergic rhinitis (AR group) and children with both rhinitis and asthma (AA group) in pulmonary function (forced expiratory volume in one second (FEV1) 100 ± 11 vs. 92.1 ± 15.0; p < 0.05). A total of 84 children (79%) presented a normal body mass index (BMI) (10-97 percentile), 8 (7.5%) were underweight, and 14 (13.5%) were overweight or obese. There were no differences in body composition between the AR and AA groups. Incorrect eating habits were demonstrated by most of the children, e.g., consumption of three or fewer meals in a day (38%), sweets every day (44%), snacking between meals every day (80%), and eating meals less than 1 h before bedtime (47%). Compared to the AR group, the AA group was more likely to eat more meals a day (p = 0.04), snack more often (p = 0.04), and eat before sleeping (p = 0.005). Multiple regression analysis showed a significant association between high BMI and snacking between meals and low physical activity (adjusted R2 = 0.97; p < 0.05). CONCLUSIONS: The risk factors for obesity in children with allergies include snacking and low physical activity. Most children with respiratory allergies, especially those with asthma, reported incorrect eating habits such as snacking and eating before bedtime. A correlation between pulmonary function and body composition or dietary habits was not found.

Cardiopulmonary exercise test and bioimpedance as prediction tools to predict the outcomes of obesity treatment.

INTRODUCTION Obesity impacts the global population. Bioelectrical impedance analysis (BIA) and cardiopulmonary exercise test [CPET]) may help modify the treatment. OBJECTIVES We aimed to compare BIA and CPET results in obese and lean indivisuals, assess changes in BIA and CPET during obesity treatment, and indentify parameters predicting treatment outcome. PATIENTS AND METHODS We enrolled 200 obese patients, of whom 45 underwent a lifestyle modification treatment, and 32 lean individuals (controls). Lifestyle modifications included diet, rehabilitation, education, and behavioral therapy. The diet was based on body composition assessed by BIA and fat metabolism assessed by CPET. The intensity of exercise in the rehabilitation program was based on CPET, mainly peak oxygen uptake (VO2peak), fat metabolism (FAT), and fat heart rate (FAT HR). The protocol duration was 12 weeks. RESULTS Obese patients differed from lean controls with regard to VO2peak (P <0.0001), oxygen uptake at anaerobic threshold (P <0.0002), respiratory exchange ratio (P <0.014), oxygen uptake to work rate slope (P <0.0004), FAT (P <0.001), FAT HR (P <0.0008), anaerobic threshold heart rate (P <0.0003), as well as fat mass (P = 0.01), fat­free mass (P = 0.007), resting metabolic rate (RMR) (P = 0.007), total body water (P = 0.01), and extracellular water (P = 0.004). The treatment resulted in increased RMR (P <0.02) and VO2peak (P <0.002), as well as reduced fat tissue (P <0.006) and resting heart rate (P <0.017). The prediction model based on FAT HR, resting heart rate, and FAT enabled the prediction of treatment outcomes in 92% of patients. CONCLUSIONS Obese patients had pathological FAT and impaired exercise tolerance. Changes in BIA and CPET prove the metabolic impact of lifestyle modification treatment.

Persistent impact of cigarette smoking on asthma.

OBJECTIVE: In the present study we assessed the impact of former cigarette smoking on asthma control and treatment effectiveness. METHODS: A total of 104 patients with uncontrolled asthma were included in the study. The group of former smokers consisted of 33 subjects, whereas the never smokers group consisted of 71 subjects of similar age and gender. Spirometry, classification of asthma severity, and control were assessed according to Global Initiative for Asthma (GINA) guidelines. Quality of life was measured with the use of the Saint George Hospital Respiratory Questionnaire (SGHRQ). RESULTS: Asthma was more severe in the group of former smokers both before and after treatment; p < 0.001. Severe asthma (OR 7.8 CI 2.8-21.9) and cigarette smoking (OR 3.5 CI 1.3-9.2) were associated with difficulties in asthma control achievement. Total quality of life significantly improved in the group of non-smokers; p = 0.02, whereas in former smokers this effect was not significant; p > 0.05. CONCLUSION: Cigarette smoking has a persistent, dose-dependent, negative impact on the response to treatment in patients with uncontrolled asthma even after smoking cessation. Smoking cessation should remain the ultimate goal in treatment of asthmatic patients. More efforts should be undertaken to decrease smoking initiation, especially in teenagers.

Differences in gene expression related to the outcomes of obesity treatment, peak oxygen uptake, and fatty acid metabolism measured in a cardiopulmonary exercise test.

INTRODUCTION The obesity pandemic requires development of methods that could be used on a large scale, such as the cardiopulmonary exercise test (CPET). Gene expression may explain CPET results on the molecular level. OBJECTIVES The aim of this study was to compare gene expression in obesity, depending on CPET results. PATIENTS AND METHODS The study group consisted of 9 obese patients and 7 controls. The treatment encompassed diet, rehabilitation, and behavioral therapy. Diet was based on the body composition analyzed by bioelectrical impedance, resting metabolic rate, and subjective patient preferences. The rehabilitation depended on the CPET results: maximal oxygen uptake and fatty acid metabolism. Behavioral intervention focused on the diagnosis of health problems leading to obesity, lifestyle modification, training in self­assessment, and development of healthy habits. The intensive treatment lasted for 12 weeks and consisted of consultations with a physician, dietitian, and medical rehabilitation specialist. RNA was isolated from the whole blood. A total of 47 323 transcripts were analyzed, of which 32 379 entities were confirmed to have high quality of RNA. RESULTS We observed differences in gene expression related to the CPET results indicating abnormalities in fat oxidation and maximal oxygen uptake. The genes with major differences in expression were: CLEC12A, HLA­DRB1, HLA­DRB4, HLA­A29.1, IFIT1, and LOC100133662. CONCLUSIONS The differences in gene expression may account for the outcomes of treatment related to inflammation caused by obesity, which affects the muscles, fat tissue, and fatty acid metabolism.

[Analysis of deaths caused by obstructive pulmonary diseases in Pomerania province in 2001-2004]. / Analiza zgonów z powodu obturacyjnych chorób pluc w latach 2001-2004 w województwie pomorskim.

INTRODUCTION: Lung diseases, especially COPD, are one of the leading causes of death. The aim of the study was to assess mortality caused by COPD, asthma and cor pulmonale in the years 2001-2004 in the Pomerania province. MATERIAL AND METHODS: Death certificates of patients who died of asthma, COPD or cor pulmonale registered in Regional Health Centre were analyzed. RESULTS: The mortality rate for COPD was 14.02/100 000, asthma 2.04/100 000 and cor pulmonale 2.4/100 000 inhabitants. Mortality rate from COPD increased from 12.5/100 000 found in 2001 to 15.6/100 000 found in 2004 (p = 0.005). The number of deaths was higher in the winter time. The peak mortality was found in patients 65 years old. 71% of COPD and cor pulmonale patients died in the hospital, whereas 75% of asthma deaths occurred at home (p = 0.0001). CONCLUSIONS: The mortality rates from chronic respiratory diseases in the analyzed population are comparable to data found in other studies. The alarming result is the increase in the number of COPD deaths and high percentage of asthma patients who die at home.

The impact of nutritional status and appetite on the hospital length of stay and postoperative complications in elderly patients with severe aortic stenosis before aortic valve replacement.

INTRODUCTION: Severe aortic stenosis (AS) is associated with the reduction of physical activity and muscle mass and may be associated with decreased appetite. AIM: To assess the nutritional status and the impact of nutritional status and appetite on the hospital length of stay and postoperative complications in elderly patients with severe AS before aortic valve replacement. MATERIAL AND METHODS: Ninety-nine patients (55 male, 44 female; 74.3 ±5.2 years old) with severe AS and an indication for aortic valve replacement (AVR) were included. The nutritional status was assessed by different questionnaires (7-point Subjective Global Assessment Score - 7-SGA, full-Mini Nutritional Assessment - full-MNA) and anthropometric measurements (body mass index (BMI) kg/m(2)). Body composition was estimated using multi-frequency bioelectrical impedance analysis. Appetite was assessed by the Simplified Nutrition Assessment Questionnaire (SNAQ). RESULTS: The average BMI of patients was 28.8 ±5.8 kg/m(2). Results of the 7-SGA and f-MNA questionnaires revealed that 39 patients (39.4%) were at risk of malnutrition. The mean SNAQ score was 15.8 ±1.8. The average length of hospital stay was 10 ±5.8 days. There was a positive correlation of LOS with age (r = 0.26, p = 0.03) and a negative correlation with fat mass (kg) (r = -0.28, p = 0.04) and BMI (r = -0.22, p = 0.03). Postoperative complications were observed in 37 patients (37.4%). Patients who developed complications were older and had poorer nutritional status according to the results of the 7-SGA. CONCLUSIONS: Despite many patients undergoing AVR being overweight and obese, a considerable proportion displayed clinical signs of malnutrition. The results suggest that an assessment of nutritional status and appetite in this group of patients should be conducted regularly and that the 7-SGA scale could represent a reliable tool to assess malnutrition.

The Role of TRAF4 and B3GAT1 Gene Expression in the Food Hypersensitivity and Insect Venom Allergy in Mastocytosis.

Mastocytosis is an uncommon disease classified as a myeloproliferative neoplasm, however, its symptoms are broad and place patients at crossroads between dermatology, hematology and allergology. Patients with mastocytosis often suffer from symptoms resulting from the activation and release of mediators from the mast cells, such as generalized itching, redness, headache, abdominal cramps, diarrhea, bone pain or arthritis, hypotension and shock. The possible severe, fatal or near fatal reactions caused by food hypersensitivity are reasons for the research focused on marker identification. The aim of the study was to analyse the gene expression differences in mastocytosis patients with and without food and drug hypersensitivity and insect venom allergy (IVA). A total of 57 Caucasian patients with mastocytosis were studied [median age 41.8; range 18-77 years; 15 (26.3 %) males and 42 (73.7 %) females]. Quantitative RT-PCRs of 11 genes plus ribosomal 18S RNA were run. Symptoms of food hypersensitivity were found in 12 patients (21 %), including 3 patients (13 %) with cutaneous mastocytosis (CM), and 9 (28 %) with indolent systemic mastocytosis (ISM). IVA was confirmed in 13 patients (22.8 %) including 6 patients (10.5 %) with CM, and 7 patients (12.3 %) with ISM. Drug hypersensitivity was diagnosed in 10 patients (17.5 %). Significant differences in the gene expression were found for TRAF4 (p = 0.008) in the comparison of the mastocytosis patients with and without concomitant food hypersensitivity. Furthermore significant differences were found in gene expression for B3GAT1 (p = 0.003) in patients with IVA compared to patients without insect sting anaphylaxis in the medical history. The expression of studied genes did not differ according to the presence of drug hypersensitivity. The TRAF4 expression was higher in mastocytosis patients with food hypersensitivity in their medical history, the B3GAT1 expression was lower in mastocytosis patients with IVA in history.

[Serum eosinophil cationic protein (ECP) in patients with perennial rhinitis and atopic dermatitis, allergic to house dust mites]. / Eozynofilowe bialko kationowe (ECP) w surowicy chorych na caloroczne alergiczne zapalenie blony sluzowej nosa i atopowe zapalenie skóry, uczulonych na roztocza kurzu domowego.

UNLABELLED: Eosinophil Cationic Protein (ECP) is released from activated eosinophils during the inflammation process. THE AIM: of the study was to evaluate levels of ECP in serum of patients with perennial rhinitis (with and without asthma) and atopic dermatitis allergic to Dermatophagoides farinae and Dermatophagoides pteronyssinus. Further it was designed to compare ECP levels in patients treated with allergen-specific immunotherapy (IT) and in patients treated symptomatically. MATERIAL AND METHODS: The study group included 94 patients allergic to house dust mite: 42 patients with perennial rhinitis not treated with IT, 24 patients with perennial rhinitis and/or asthma treated with IT and 28 patients with atopic dermatitis. In the control group were 21 healthy volunteers. In the group treated with IT questionare of efficacy and safety was performed. RESULTS: The serum level of ECP was measured using immunofluorometric assay. In the group with perennial rhinitis treated with IT mean level of ECP was 6,5 ug/l, whereas in the group not treated with IT--15,78 microg/l (p=<0.05). In patients with atopic dermatitis ECP level was the highest--23,04 microg/l +/- 4,98 and was significantly different than in the group of healthy volunteers-- 7,2 microg/l +/- 1,1 (p=0,0048). CONCLUSIONS: Serum ECP concentration may be prognostic factor in specific immunotherapy.

[Systemic mastocytosis]. / Mastocytoza ukladowa.

Mastocytosis is group of rare disorders characterized by abnormal mast cells growth. Pathology of the disease is unknown, mechanisms involved in mast cells delineation and growth are considered to play an important role. Unusual and broad spectrum of symptoms therefore requires cooperation of different specialists. The aim of the study was to assess diagnostic and therapeutic methods used in the Gdansk Mastocytosis Centre. 14 patients were studied (9 adults and 5 children). Bone marrow biopsy was performed in order to assess the stage of disease. Pathological, cytological, cytofotometry and genetic examination (C-KIT mutation) of the bone marrow were performed. Patients suffereing from food allergy and wasp venom anaphylaxis were diagnosed with skin prick tests and sIgE. All subjects suffered from urticaria pigmenthosa and anaphylaxis. Indolent mastocytosis was diagnosed in six subjects. Patients were treated with antihistamines, corticosteroids, cromones and immunotherapy with a marked reduction of symptoms. The experience of Gdansk Mastocytosis Centre indicates that mastocytosis is a rare and difficult to diagnose disease.

Analysis of the differences in whole-genome expression related to asthma and obesity.

INTRODUCTION: Concomitant obesity significantly impairs asthma control. Obese asthmatics show more severe symptoms and an increased use of medications. OBJECTIVES: The primary aim of the study was to identify genes that are differentially expressed in the peripheral blood of asthmatic patients with obesity, asthmatic patients with normal body mass, and obese patients without asthma. Secondly, we investigated whether the analysis of gene expression in peripheral blood may be helpful in the differential diagnosis of obese patients who present with symptoms similar to asthma. PATIENTS AND METHODS: The study group included 15 patients with asthma (9 obese and 6 normal-weight patients), while the control group-13 obese patients in whom asthma was excluded. The analysis of whole-genome expression was performed on RNA samples isolated from peripheral blood. RESULTS: The comparison of gene expression profiles between asthmatic patients with obesity and those with normal body mass revealed a significant difference in 6 genes. The comparison of the expression between controls and normal-weight patients with asthma showed a significant difference in 23 genes. The analysis of genes with a different expression revealed a group of transcripts that may be related to an increased body mass (PI3, LOC100008589, RPS6KA3, LOC441763, IFIT1, and LOC100133565). Based on gene expression results, a prediction model was constructed, which allowed to correctly classify 92% of obese controls and 89% of obese asthmatic patients, resulting in the overall accuracy of the model of 90.9%. CONCLUSIONS: The results of our study showed significant differences in gene expression between obese asthmatic patients compared with asthmatic patients with normal body mass as well as in obese patients without asthma compared with asthmatic patients with normal body mass.

Risk factors for anaphylaxis in patients with mastocytosis.

INTRODUCTION: Symptoms resulting from the activation and release of mediators from the mast cells are observed in about 30% of the patients with mastocytosis. OBJECTIVES: The aim of the study was to assess the prevalence of anaphylactic reactions and to identify the risk factors for anaphylaxis in patients with mastocytosis depending on the type of the disease. Furthermore, we analyzed a response to treatment of mediator-related symptoms in this patient group. PATIENTS AND METHODS: The study group included 152 adult patients with mastocytosis. The diagnostic workup included a histopathological examination, flow cytometry, KIT mutation analysis, and measurement of tryptase levels. The diagnosis of allergy was confirmed by the skin prick test and serum immunoglobulin E levels. RESULTS: The prevalence of anaphylactic reactions in the study group was 50% and was higher in patients with systemic mastocytosis (P = 0.007), specifically in its indolent variant (P = 0.026), than in patients with cutaneous mastocytosis. The most frequent triggers of anaphylaxis were food (29%), insect stings (22%), and drugs (15%). Tryptase levels were higher in patients with a history of anaphylaxis (P = 0.029) as well as in those with symptoms provoked by physical factors (P = 0.002). Such symptoms were reported in 112 patients (74%) and were more common in patients with systemic mastocytosis compared with those with cutaneous mastocytosis (P = 0.026). The treatment was ineffective in 8 patients (10.5%) and resulted only in partial remission in 14 patients (18.4%). CONCLUSIONS: The study showed a significant incidence of symptoms related to physical factors in patients with mastocytosis and anaphylaxis in history. Risk factors for anaphylaxis included increased serum tryptase levels and indolent variant of systemic mastocytosis. Standard pharmacological treatment was ineffective in 10% of the patients, who may require biological treatment.  

Gene expression analysis in allergology: the prediction of Hymenoptera venom allergy severity and treatment efficacy.

Insect venom allergy (IVA) may result in the most severe systemic reactions seen in allergology. The only potentially curative treatment option is venom immunotherapy (VIT) over 3 to 5 years. This treatment is effective in more than 90% of subjects but no reliable predictors of VIT effectiveness exist. Sting challenge with a living insect can be performed to assess the effectiveness of VIT: the predictive value of sting challenge can be highly sensitive in patients with honeybee venom allergy whereas in yellow jacket allergy, a negative result can be reliable if the challenge has been repeated at least 3 times.The analysis of gene expression may be a step towards personalized venom immunotherapy assessing the effectiveness of treatment, the minimal required time for VIT and the persistence of long term tolerance induced by the treatment. Recent studies have enabled construction of a predictive model that could potentially be used in clinical practice to assess the efficacy of insect venom immunotherapy. A set of 69 genes that may be responsible for long-term protection was identified. Further analysis of the previously identified 6 transcripts make up the 18 gene predictive peripheral blood showed differences in patients treated with IVA. Further studies are needed to investigate the usefulness of gene expression analysis and other markers in the prediction of VIT effectiveness.