Development of a high-resolution melting analysis method based on SYBR green-I for rs7216389 locus genotyping in asthmatic child patients

Asthma is caused by the combination of different factors. Current concepts of asthma pathogenesis emphasize on gene-environment interactions. Mega-genome scanning projects revealed that different Single Nucleotide Polymorphisms [SNPs] are related to asthma susceptibility. rs7216389-T is one of them that is related to childhood asthma and its effect on childhood asthma severity has been proved in different nations, however no study has been performed in Eastern Mediterranean and Middle East countries yet. To perform population genetic studies, a rapid and high-throughput screening method is necessary. High-resolution melting analysis is a rapid, powerful and accurate method, which is suitable for this type of studies. Therefore, it has been decided to develop a high-resolution melting method for rs7216389 locus genotyping in Iranian asthmatic children. In the current study, a high-resolution melting analysis method based on SYBR Green-I was developed to check the frequency of rs7216389-T mutation in Iranian asthmatic children for the first time. Second and third classes of intercalating dyes are commonly used for high-resolution melting method. However, in this study, SYBR Green-I was used for rs7216389 locus genotyping for the first time. Our results for 60 samples showed that SYBR Green-I has good efficacy for rs7216389 locus genotyping through high-resolution melting method in comparison with PCR-RFLP and sequencing. Comparison of our results based on HRM analysis with PCR-RFLP showed that our developed method is rapid, accurate, high-throughput and economic to study the rs7216389 locus in asthmatic children and it is applicable for other similar population genetic studies

survey of pediatricians' knowledge on asthma management in children

Asthma is one of the most common acute and chronic conditions in children, and the pediatricians are expected to provide an important role for asthma care in this age group, however there is no published information describing the different aspects of their practices about children asthma in Iran. This study was done to characterize the knowledge of the Iranian pediatricians about the diagnosis, treatment and education of asthma in children. Validated questionnaires were completed by 193 pediatricians from different parts of Iran during the International Congress of Pediatrics in Tehran. A total of 193 returned questionnaires [96.5%] were eligible for the survey and analysis. About 49% of the respondents were male and 18% were sub-specialists. Wheezing was the most common mentioned symptoms in taking asthma into consideration. About 40% of these physicians had no plan for doing spirometry in eligible children and 35.2% of them did not have familiarity with peak flowmeter. Also about 17.6% of them paid no regular visits to their asthmatic patients. Only 29% of the respondents indicated that they would prescribe inhaled corticosteroids for a 6-year-old child with moderate persistent asthma and 73.3% of them would prescribe inhaled bronchodilator [Salbutamol] for an acute asthmatic attack as the first drug, while 17.1% of them used epinephrine injection for this purpose. About 42.2% of the respondents did not consider any education or action plan for their patients and only 60.6% of them had access to standard guidelines and educational programs. The results show that there are numerous aspects of children asthma management in Iran which are not consistent with standardized guidelines and recommendations. This survey and the attained information suggest areas for interventions to improve the pediatricians' knowledge about asthma and the disease management

Autoimmune progesterone anaphylaxis

Progesterone induced dermatitis is a rare disorder. It typically occurs in females due to an autoimmune phenomenon to endogenous progesterone production, but can also be caused by exogenous intake of a synthetic progestin. Here in, we present a case of autoimmune progesterone anaphylaxis [AIPA] observed in an adolescent female. The patient is an 18-year-old Caucasian female with no significant past medical history and no prior exogenous hormone use, who presented to her primary care physician complaining of cyclic skin eruptions with dyspnea, cough and respiratory distress. She noted that her symptoms occurred monthly, just prior to her menses. An intradermal skin test using 0.1 cml of progesterone was performed. The patient developed a 15mm wheal after 15 minutes, confirming the diagnosis of AIPA. The patient was started on a continuous regimen of an oral conjugated estrogen [0.625mg]. The skin eruptions and respiratory symptoms have not returned since the initiation of this therapy. Autoimmune progesterone dermatitis manifests via the occurrence of cyclic skin eruptions. Women with the disorder commonly present with dermatologic lesions in the luteal phase of the menstrual cycle, if there are any other organ involvement in addition to skin [e.g. lung, GI] the reaction should be called as autoimmune progesterone anaphylaxis. Diagnosis of AIPA is confirmed by performing a skin allergen test using progesterone

Systemic and local reactions of bee venom immunotherapy in Iran

Severe allergic reactions during specific immunotherapy may occur in the treatment of hymenoptera sting allergy. The objective of the present study was to examine the characteristics of allergic reactions during specific immunotherapy in patients with allergy towards hymenoptera venom in the Iranian population. A prospective study was performed using the clinical reports of 27 patients with anaphylaxis to bee venom [Apis melifera, Geupes vespula and Geupes Polites]. Ten patients treated with Cluster protocol during 2002 and 2006 After diagnosis of hymenoptera sting allergy according to history and intradermal tests, the patient were treated with Cluster protocol immunotherapy. The protocol lasted 6 weeks with an increase in the concentration of venom from 0.01? g/ml to 100 ? g/ml. None of the patient received premedication. All patients with hymenoptera venom allergy received 120 injections. Anaphylactic reactions were classified according to the Mueller-classification. The frequencies of systemic reactions during Cluster protocol were 8.33% and 5% for yellow jacket and honey bee venom respectively. No patient experienced severe systemic reaction. Cluster protocol for hymenoptera immunotherapy is a reliable method for the treatment of anaphylactic reactions to bee venom. It is safe with low cost and do not need hospitalization

High doses intravenous immunoglobulin versus oral cyclosporine in the treatment of severe atopic dermatitis

Atopic dermatitis is one of the most common allergic diseases that almost always respond to conventional therapies with topical emollient, topical corticosteroids, systemic antihistamines and allergic abstinence. However few cases of atopic dermatitis with severe course do not respond to conventional therapies and high dose of intravenous immunoglobulin or cyclosporine are recommended for them. This clinical trial study has been done to compare the last two regimens in patients with severe atopic dermatitis, Scoring Atopic Dermatitis [SCORAD] > 70. We included 14 patients in two groups. In group 1, eight patients were randomly selected and received 4mg/kg cyclosporine daily for 3 months and in group 2, six patients received 2g/kg Intravenous Immunoglobulin [IVIG] as stat infusion. All patients were followed on days 15, 30, 60 and 90 after starting the therapy. About 75% and 62.5% of patients had positive skin tests to egg and to milk respectively. Six patients out of 14 patients did not have skin test, so specific IgE by Radioallergosobent tests [RAST] was used for them. All of these patients had positive RAST to egg and 66.6% against cow's milk. There was a significant difference in the clinical outcomes of these two groups with a marked reduction in SCORAD of day 90[th] in group 1 in comparison to group 2 [Pvalue = 0.005]. No significant adverse drug reaction was seen in these two groups

Total serum igE and skin tests in children with respiratory allergy

Total serum IgE measurement and skin prick tests are the most common tools for allergy diagnosis. This investigation was conducted to evaluate total serum IgE and skin prick test relationship in Iranian atopic children with respiratory manifestations. A history of allergic rhinitis, bronchial asthma and atopic dermatitis was obtained in 232 children [aged 1 to 15 years]. Skin prick testing was performed using commercial preparation of the common allergens. The patients'sera were also analyzed for determining total IgE. More than 95% [n=221] of the patients had positive skin prick test [mean wheal diameter > /=3mm] to one allergen or more. Among the patients 46% [n=107] had elevated total serum IgE levels [> /=150 IU/ml]. The study showed close relationship of skin test positivity with reported allergic symptoms, but no correlation was found between total IgE and skin prick test. This study revealed that skin testing and total serum IgE measurement may be considered complementary to one another in diagnosing allergic respiratory disorders

Neutropenia in patients with primary antibody deficiency disorders

Neutropenia is characterized by decrease in the absolute number of circulating neutrophils and an increase susceptibility to infections. The current study was performed in order to explain the clinical and laboratory findings of patients with antibody deficiency disorders associated neutropenia. The patients' records of 19 neutropenic cases out of 207 patients with antibody deficiencies, who had been referred to Children's Medical Center and enrolled in Iranian primary immunodeficiency registry, were reviewed. Nineteen cases [14 male and 5 female], with a mean age of 10.7 +/- 5.7 years, were associated with neutropenia [9.2%]. The disorders with associated neutropenia were Hyper IgM syndromes [3 of 8], Common variable immunodeficiency [13 of 109], and X-linked agammaglobulinemia [3 of 45]. The median age for the onset of disease and diagnosis age were 15 months [1-134] and 3.8 years [6 months-13 years], respectively. The most common infections during the course of illness were pneumonia [13 cases], diarrhea [12 cases], oral candidiasis [9 cases], otitis media [6 cases], sinusitis [6 cases], cutaneous infections [5 cases], and abscess [5 cases]. Other less frequent infections were: conjunctivitis, oral ulcers, meningitis, and osteomyelitis. Three neutropenic patients died because of recurrent infections. Neutropenia may occur in any of the primary immunodeficiency disorders. Persistent or severe infections always pose a supposition, which deserves further evaluation for detecting an underlying immune deficiency syndrome and neutropenia, since a delay in diagnosis may result in a serious organ damage or even death of the patient