Influence of the Polymorphism C-509T in the TGFB1 Gene Promoter on the Response to Montelukast.

Transforming growth factor beta 1 (TGFB1) is a multifunctional cytokine with a key role in asthma airway inflammation and remodeling. Since elevated levels of this cytokine in airways might be associated with response to asthma therapy, the aim of this study was to investigate whether the presence of the polymorphism C-509T in the promoter of the TGFB1 gene is associated with response to montelukast. A group of 102 asthmatic patients was genotyped for the presence of the C-509T polymorphism by DNA sequencing and subjected to induced sputum sampling. Cells from sputum samples and BEAS 2B cells were treated with montelukast and endogenous TGFB1 expression was measured by quantitative real-time polymerase chain reaction. The promoter activity was analyzed by luciferase assays in BEAS 2B cells transfected with constructs carrying variants -509C and -509T of the TGFB1 gene promoter. After treatment with montelukast, the decrease in TGFB1 gene expression was greater for the -509TT genotype (58.9%) than for the -509CC and -509CT genotypes (49.6% and 31.8%, respectively) (P = 0.071). In BEAS 2B cells, expression of endogenous TGFB1 was reduced by about 27% after montelukast treatment, while luciferase activity of both promoter variants was increased after montelukast treatment (-509C allele: 48.3%, P = 0.060; and -509T allele: 100.5%, P = 0.062). A more intensive response was registered in the promoter containing the -509T allele, which had 135% higher activity than the -509C variant (P = 0.035). This study showed that the presence of the -509T allele in the TGFB1 promoter might modulate effects of montelukast on TGFB1 gene expression, but future studies are necessary, taking into consideration other genetic and nongenetic factors. It is of potential importance for clinical management of asthma to clarify the influence of the C-509T polymorphism on the response to treatment with montelukast.

Neutrophil elastase gene polymorphisms: modulators of response to therapy in childhood bronchiectasis?

BACKGROUND: The aim of this study was to investigate polymorphisms in the promoter region of the neutrophil elastase (ELANE) gene as potential modulators of the therapeutic response in children with idiopathic bronchiectasis. METHODS: The study included 48 children between 5 and 17 years old who were diagnosed with idiopathic bronchiectasis based on high-resolution computed tomography of the thorax. In all patients therapy included administration of antibiotics, anti-inflammatory drugs, expectorants, and postural drainage. Response to therapy was evaluated by the change in FeNO levels before and after administration of therapy. The ELANE promoter region polymorphisms were analyzed by PCR-direct DNA sequencing. RESULTS: According to the predicted activity of ELANE genotypes, subjects were divided into two groups: low/intermediate activity (n = 18) and high activity (n = 30). Subjects in the group with high-activity genotype had higher initial FeNO levels and this difference was statistically significant (t = 2.906; p = 0.006). The difference between FeNO levels before and after therapy was also statistically significantly higher in children with high-activity genotype (t = 3.329; p = 0.002). Statistically significant correlation was observed between the change in FeNO levels and ELANE genotypes (r = 0.350; p = 0.015). CONCLUSION: Children with high-activity genotype had higher initial FeNO levels and showed better response to therapy than children with low/intermediate-activity genotypes.

Complications of mechanical ventilation in pediatric patients in Serbia.

BACKGROUND: Mechanical ventilation is a frequently applied therapy in critically ill children and can be lifesaving in many cases. Clinical use of this technique has well documented benefits, but can be associated with different complications and adverse physiologic effects. OBJECTIVES: The aim of this study was to investigate the complications and clinical outcome of mechanical ventilation in Serbian pediatric patients. MATERIAL AND METHODS: The study encompassed 42 children with respiratory insufficiency that underwent mechanical ventilation during hospitalization over a period of 12 consecutive months. The influence of clinical and mechanical parameters on the occurrence of complications and clinical outcome were analyzed. RESULTS: The patients were ventilated for a total of 432 days and a total of 61 complications were observed in 42 patients (97 complications per 1000 ventilation days). The most common complications associated with mechanical ventilation in Serbian pediatric patients with respiratory insufficiency were cardiovascular insufficiency (52.4%) and multiple organ failure (35.7%). High values of applied PIP (> 26 cm H 2 O), PEEP (> 6 cm H 2 O) and Tv (> 6 mL/kg) were associated with the occurrence of complications and negative clinical outcome. CONCLUSIONS: Complications of mechanical ventilation in the pediatric population occur frequently, but lower volumes/pressures of ventilation contribute to a decrease in incidence. Further studies are needed to examine associated risk factors and strategies to reduce the occurrence of complications and improve clinical outcome.

Anaphylaxis followed by unilateral lung opacity and hypocomplementemia in a young female.

A 36-year-old woman was stung in the right wrist by a bee, suffered typical anaphylaxis, and was transferred to a local hospital. After a few hours, which corresponded to late-onset reaction, she developed shortness of breath and weakness and was transferred to the emergency department, where the diagnosis of anaphylaxis was confirmed. Serum complement levels, components C3 and C4, were undetectable. Flexible bronchoscopy excluded lung hemorrhage. She was on the ventilator for 4 days and was fully awake during that time. After the treatment, her improvement was rapid, and she fully recovered. Three weeks after she had been stung by a bee, skin prick tests to bee, wasp, and yellow jacket venom were done, and all tests came back negative as well as IgE and IgG4 antibodies to the same venoms. Coagulation factors and the complement, including C1q inhibitor, were normal. Occurrence of complement activation and consumption could point to the immune complexes as basis of pathophysiological mechanism. It remains unclear why such a reaction would involve only the right lung and why no detectable immune complexes were discovered. The clinical picture in the presented case resembled acute respiratory distress syndrome, but the exact nature of lung consolidation remains puzzling. The most likely explanation is a very rare case of airway obstruction. To our knowledge, this is the only reported case of anaphylaxis associated with undetectable serum complement levels. The potential role and diagnostic significance of hypocomplementemia in cases of anaphylaxis should be further investigated.

Analysis of CFTR Gene Variants in Idiopathic Bronchiectasis in Serbian Children.

This study has investigated a potential role of common Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene variants in the etiology of noncystic fibrosis bronchiectasis in Serbian children. The study has encompassed 48 patients (19 male and 29 female, aged between 5 and 18 years, median age 10.6±3.3), diagnosed with idiopathic bronchiectasis based on high-resolution computed tomography of thorax and pathologic examination of lobectomy materials. The CFTR gene analysis was performed on genomic DNA extracted from peripheral blood samples of patients by polymerase chain reaction (PCR)-Mediated Site-Directed Mutagenesis method, Denaturing Gradient Gel Electrophoresis method, and DNA sequencing. Mutation c.1521_1523delCTT (F508del) was detected with an allelic frequency of 1.0%, and c.224G>A (R75Q) variant. Carriers of c.1210-12T[5] (IVS8-5T) allele were significantly more common than in the general population (10.4% vs. 5.0%, P=0.0302). The frequency of homozygotes for Met 470 allele was higher in patients than in the general population (33% vs. 20%), while heterozygotes for p.Met470Val were less frequent (31% vs. 50%), and this difference was statistically significant (P=0.0222). The results obtained in this study indicate involvement of 2 common CFTR variants, c.1210-12T[5] and c.1408A, in idiopathic bronchiectasis in children, but this observation should be further confirmed by more extensive analysis of the CFTR gene in a larger group of patients.

Presentation of an infant with nutritional deficiency dermatitis as the initial manifestation of cystic fibrosis.

INTRODUCTION: Cystic fibrosis (CF) is a multisystemic autosomal recessive disease most frequently recognized by characteristic respiratory and/or digestive manifestations. Exceptionally rare, as is the case with the infant we are presenting, the initial sign of the disease can be nutritional deficiency dermatitis (NDD). CASE OUTLINE: A three-month-old male infant of young and healthy non-consanguineous parents, born at term after the first uneventful pregnancy, was hospitalized due to atopic dermatitis (AD)-like skin changes, failure to thrive and normochromic anemia (Hb 60 g/L). As exclusively breast-fed, failure to thrive was attributed to hypogalactia and skin changes to nutritional allergy, so that, besides exclusion of cow's milk protein and other highly allergenic foods in mother's diet, hypoallergenic milk formula was added to the child's diet. However, dietetic measures were without effect, and the child was re-hospitalized at age 4.5 months, this time in the condition of severe malnutrition with hypoproteinemic edemas, extensive dermatitis, moderate hepatosplenomegaly and recurrent normochromic anemia (Hb 57 g/L). After plasma-free erythrocyte transfusion, correction of hypoalbuminemia and two-week parenteral and semi-elementary nutrition resulted in gradual recovery of the child, also including the resolution of skin changes. Having in mind the clinical course of the disease, as well as the response to applied therapeutic measures, CF was suspected as the cause of the child's problems, which was also confirmed by a high level of sweat chlorine (92 mmol/L) and DNA analysis (deltaF508/deltaF508). CONCLUSION: Our experience indicates that NDD, as the initial manifestation of CF, should be also kept in mind in differential diagnosis of the infant's AD-like changes.

Immediate allergic reaction to methylprednisolone with tolerance of other corticosteroids.

INTRODUCTION: In spite of the wide usage of corticosteroids for the treatment of a plethora of diseases, sometimes they can induce immediate hypersensitivity reactions, which are however uncommon. CASE OUTLINE: We report a case of immediate allergic reaction induced by intravenous methylprednisolone given before operation for surgical repair of an arm contracture as a sequel of burns, which the child had tolerated a month before. Six weeks later the patient repeated the anaphylactic reaction during skin testing to methylprednisolone. In addition, basophile activation test with methylprednisolone (BAT) was positive. CONCLUSION: This case report describes a patient who experienced intraoperative anaphylaxis and anaphylactic reaction induced by skin testing. This is the first report on induction of both anaphylactic reactions by methylprednisolone in the same child. Clinical findings, positive BAT and positive skin tests with methylprednisolone imply that the child developed type-I hypersensitivity. The lack of crossreactivity with other corticosteroids emphasizes that the reactions were caused by the steroid molecule.

[Kiss-induced severe anaphylactic reactions].

INTRODUCTION: Ingestion is the principal route for food allergens to trigger allergic reaction in atopic persons. However, in some highly sensitive patients severe symptoms may develop upon skin contact and by inhalation. The clinical spectrum ranges from mild facial urticaria and angioedema to life-threatening anaphylactic reactions. OUTLINE OF CASES: We describe cases of severe anaphylactic reactions by skin contact, induced by kissing in five children with prior history of severe anaphylaxis caused by food ingestion. These cases were found to have the medical history of IgE mediated food allergy, a very high total and specific serum IgE level and very strong family history of allergy. CONCLUSION: The presence of tiny particles of food on the kisser's lips was sufficient to trigger an anaphylactic reaction in sensitized children with prior history of severe allergic reaction caused by ingestion of food. Allergic reaction provoked with food allergens by skin contact can be a risk factor for generalized reactions. Therefore, extreme care has to be taken in avoiding kissing allergic children after eating foods to which they are highly allergic. Considering that kissing can be a cause of severe danger for the food allergic patient, such persons should inform their partners about the risk factor for causing their food hypersensitivity.

Novel cftr gene sequence variation in Serbian patient with idiopathic disseminated bronchiectasis.

This paper reports a novel Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene variant, 1811+1G-->T, detected in a 5-year-old girl diagnosed with idiopathic disseminated bronchiectasis and negative sweat chloride test (17 mmol/L). The performed CFTR gene mutation analysis included detection of the F508del mutation, analysis of Tn polymorphism and screening of CFTR exons 3, 10 and 11. The CFTR gene screening has shown the altered band pattern in exon 11. The DNA sequencing of CFTR exon 11 revealed the presence of the novel sequence variation 1811+1G-->T in heterozygous state. This sequence variation was not found in any of 100 control alleles, analyzed by polymerase chain reaction - restriction fragment length polymorphism method. The novel sequence variation 1811+1G-->T is located at the splicing site at the boundary of exon 11 and intron 11 and might be either a sequence variation or a splicing site defect.

Immediate allergic reactions to cephalosporins and penicillins and their cross-reactivity in children.

Penicillins and cephalosporins are the most important betalactams inducing IgE-mediated reactions. The safety of administering cephalosporins to penicillin-allergic children is a particular problem, because cephalosporin allergenic determinants have not been properly identified. A study was undertaken to evaluate the frequency of anaphylactic reactions to cephalosporins and penicillins and their cross-reactivity in a pediatric population. A prospective survey was conducted in a group of 1170 children with suspected immediate allergic reactions to cephalosporins and/or penicillins, which were examined during a period of 8 yr. In vivo (skin tests and challenges) and in vitro tests (for specific IgE) were performed with standard concentration of penicillins and cephalosporins. When 1170 children with a clinical history of allergy to penicillins and/or cephalosporins were tested in vivo for immediate hypersensitivity to betalactams, 58.3% cases overall were found to be skin or challenge test positive. Among them, 94.4% patients were positive to penicillins and 35.3% to cephalosporins. The frequency of positive reactions in the in vivo testing was in the range from 36.4% to 88.1% for penicillins and from 0.3% to 29.2% for cephalosporins. However, 31.5% of the penicillin allergic children cross-reacted to some cephalosporin. If a child was allergic to a cephalosporin, the frequency of positive reactions to penicillin was 84.2%. The cross-reactivity between cephalosporins and penicillins varied between 0.3% and 23.9%. The cross-reactivity among different generations of cephalosporins varied between 0% and 68.8%, being the highest for first and second-generation cephalosporins and 0% for third generation cephalosporins. The frequency of immediate allergic reactions to cephalosporins is considerably lower compared to penicillins, and the degree of cross-reactivity between cephalosporins and penicillins depends on the generation of cephalosporins, being higher with earlier generation cephalosporins. The cross-reactivity among cephalosporins is lower compared to cross-reactivity between penicillins and cephalosporins.

[Value of a rapid test for identification of beta-hemolytic streptococcus antigens in children with streptococcal pharyngitis]. / Vrednost testova za brzu identifikaciju antigena beta-hemolitickog streptokoka kod dece sa streptokoknim faringitisom.

Beta-hemolytic group A streptococcus (Streptococcus pyogenes) is the most common bacterial agent associated with the upper respiratory tract infections in humans. The most frequently group A streptococcus-associated disease is pharyngitis. Males and females are equally affected by group A streptococcus. There is seasonal increase in the prevalence of group A streptococcus-associated pharyngitis. Streptococcal pharyngitis is most prevalent in winter and early spring with higher incidence of disease observed in crowded population such as school children. Early diagnosis and treatment of group A streptococcal pharyngitis has been shown to reduce the severity of symptoms and further complications such as rheumatic fever and glomerulonephritis. The conventional methods used for identification of group A streptococci depend on isolation and identification of the organism on blood agar plates. These methods usually require 18-24 hours of incubation at 37 degrees C. Such delay in identifying the group A streptococcus has often made physicians to administer therapy without first disclosing the etiological agent. Development of immunologic tests, capable of detecting the group A streptococcal antigen directly from the throat swabs, produced rapid test results employed for better treatment of patients. STREP A test is a rapid immunochromatographic test for the detection of group A streptococci from throat swabs or culture. The accuracy of the test does not depend on the organism viability. Instead, group A strep antigen is extracted directly from the swab and identified using antibodies specific for the group A carbohydrates. We compared rapid test with conventional throat swab in 40 children, who met Centor criteria for streptococcal pharyngitis (absence of cough, high fever, purulent pharyngitis, enlarged and painful cervica lymph nodes). Overall congruence of rapid test and culture was 94%. Test is easy to perform and it is recommended as the first diagnostic test for management of children with streptococcal pharyngitis. In children with negative test, but with characteristics highly suggestive of streptococcal infection, throat culture should be performed.

[Allergy to cephalosporin antibiotics in childhood]. / Alergija na cefalosporinske antibiotike u decjem uzrastu.

A particular problem is the safety of administering cephalosporins to penicillin-allergic children, because cephalosporin allergenic determinants have not been properly identified. Cephalosporin antibiotics are widely used to treat common infections and are often the first-line prophylaxis before many types of surgery. So the arm of this study is to determine the frequency of allergic reactions of anaphylactic type to cephalosporins and their cross-reactivity with penicillins. At University Children's Hospital in Belgrade a group of 1,170 children with suspected anaphylactic allergic reaction to penicillins and/or cephalosporins were tested for the last eight years. Skin tests were performed with standard concentration of penicillins and cephalosporins. In children where skin tests were negative single-blind, placebo-controlled challenges were performed. In case of positive skin tests further examinations were interrupted and the children were considered allergic to that drug. The frequency of anaphylactic allergic reactions to cephalosporins is 0.2% to 17%, and depends on cephalosporins generation. The cross-reactivity between cephalosporins and penicillins is 0.1% to 14.5%, and among cephalosporins is 0% to 11.7%.

[Desensitization to diphtheria, tetanus and pertussis vaccine]. / Hiposenzibilizacija vakcinom protiv difterije, tetanusa i pertusisa.

Immunization with DTP vaccine (diphtheria, tetanus and pertussis) is a part of the vaccination calendar offered in childhood. Adverse allergic reactions vary from minimal urticarial reactions to life-threatening anaphylaxis. In infancy these reactions usually interrupt the vaccination calendar, but immunization in these children should be done. At the University Children's Hospital of Belgrade, a group of 137 children with suspected allergic anaphylactic reaction to DTP, DT, TT and monopertussis vaccine was studied for the last six years. Skin (prick and intradermal) tests were performed with corresponding vaccine. If both tests were negative, the vaccine could be given as a single dose of 0.5 ml. If one of these tests were positive desensitization with vaccine could be done (according to the protocol described by Carey and Meltzer). In one group of 52 children three days before desensitization, premedication with antihistamines, was done, whereas in the other group of 52 children premedication was not done. Two (3.8%) children in a group of 52 children with premedication had a minor (local) reaction after vaccination and 50 children (96.2%) had no reaction after vaccination, whereas no children (0%) had systemic reaction after desensitization.