Sequencing and expression of elastase 11 gene in polymorphoneuclear leukocytes in healthy individuals for determination of single nucleotide polymorphism [SNP] by RT-PCR

ELA2 gene responsible for coding human neutrophil elastase, a powerful serine protease carried by blood neutrophils and capable of destroying most connective tissue proteins. The NE gene consist of 5 exons and 4 introns. Peripheral blood obtained from healthy individuals. Total RNA was isolated using RNA standard techniques from fresh separated cells by polymorphoprep. RNA was analyzed by employing PCR amplification of reveres transcribed using a total of ten specific primers. We amplified five exon of ELA2 gene separately and sequenced each exon. Mutational analysis was performed by directed capillary sequencing method. We have found new single nucleotide polymorphism [SNP] in exon II codon 44. It was a silent mutation G to A substitution but no changes in amino acid sequences were seen. The codon sequence was GCG that has changed to GCA. The purpose of SNP detection is mainly recognition of susceptible people to certain diseases and providing suitable drugs based on each person genetic information. SNP information in Iranian patients can be used for analysis of drug response to human diseases

Common food allergens in Iranian children

The prevalence of food allergy is different in different parts of the world. Identification of the most common food allergens is a priority in any population to provide effective preventive and curative measures. The aim of this study was to determine the most common food allergens in Iranian children. One hundred and ninety children with skin, respiratory or gastrointestinal symptoms, thought to be due to food allergy, were studied. The total serum IgE and eosinophil count tests were measured in all patients. Allergy to 25 food allergens was determined according to the patient's history, skin prick tests, RASTs [Radioallergosorbent Test] and open food challenge tests. Findings: The most common food allergens were cow's milk, tomato, egg white, egg yolk, beef and almond, respectively. The order of common food allergens in this study was different from other reports; this might be due to the different food habits and /or ethnic diversities

study of cockroach allergy in Iranian children with asthma

Asthma is the most common chronic disease of childhood. Exposure to allergens plays an important role in asthma. Recently, cockroach has attracted attentions as an important allergen. To evaluate cockroach allergy in Iranian children with asthma. 100 children with asthma were studied. After determination of asthma severity, serum total IgE and blood eosinophils were measured. The patients were then skin prick tested with cockroach [Blattella germanica] and some common aeroallergens and food allergens. According to cockroach skin prick test [SPT] results, the patients were divided into two groups [sensitive and non-sensitive] and compared. Positive SPT for cockroach was detected in 29% of patients. Cockroach sensitive group had a more severe asthma [p<0.02]. Asthmatic patients with cockroach allergy had perennial symptoms [p<0.05]. A remarkable number of sensitive patients were born in winter [p<0.05]. Eosinophilia was more common in sensitive group [p<0.02] and they had a higher level of serum total IgE [p<0.04]. The prevalence of cockroach allergy was near to other common aeroallergens. Cockroach sensitive group had perennial symptoms, which are compatible with other indoor allergens. Cockroach allergy was more prevalent in children born in winter. May be these children are kept at home during their first months of life and are more exposed to cockroaches. The higher prevalence of eosinophilia and higher IgE level in sensitive group may be the reasons of more severe asthma in this group

20-year survey of infectious complications in 64 patients with common variable immunodeficiency

Common variable immunodeficiency [CVID] is a heterogeneous primary immunodeficiency disorder, characterized by hypogammaglobulinemia and increased susceptibility to recurrent bacterial infections. To determine the spectrum of infectious complications in patients with common variable immunodeficiency [CVID], we reviewed the hospital records of 64 CVID patients, who were diagnosed in Children's Medical Center during a period of 20 years. Among our patients, there were 38 males and 26 females, with a median age of 12 years [2-42 years] at the time of study. The median age of diagnosis was 6.1 years, with an average diagnostic delay of 5.2 years in our patient's group. Almost all of our patients have suffered from acute and recurrent infections, particularly in the respiratory and gastrointestinal systems. The majority of patients [82.5%] had pneumonia prior to diagnosis. The other infectious complications seen in our patients were as follows: recurrent otitis media [54%], recurrent sinusitis [50.8%], diarrhea [65.1%], and bacterial meningitis [11.1%]. Unusual or opportunistic infections were also seen in some of our patient population, including oral candidiasis in 10 patients, and Pneumocystis carinii pneumonia in 2 subjects. Based on this study, we suggest that hypogammaglobulinemia should be considered in any patient with a history of recurrent infections in different organs, and such patients should have a full assessment of the immune system

BCG dissemination in 40 patients and the review of leukocyte mycobacterium deffect in one patient

We have reviewed the medical records of 40 patients with disseminated [Bacille Calmette-Guerin] BCG from 1996 to 1999 in the Immunology Department of the Children Hospital, Medical Center, Tehran University. These patients are divided in to 3 groups: 1. patients who had disseminated [Bacille Calmette-Guerin] BCG after vaccination and their diagnosis was chronic granulomatous disease. 2. Patients who had disseminated [Bacille Calmette-Guerin] BCG and were diagnosed as having cell mediated immunodeficiency. 3. patients whose Nitroblue Tetrazulium and CMI were around normal, but they could not kill intracellular mycobacterium, because of confirmed deficiency of interferon-gamma receptor and IL[12] receptor. Interferon-gamma receptor or CD119 was checked in 6 patients. In one patient interferon-gamma receptor deficiency was confirmed by flowcytometric analysis. In other patients, this marker was around normal, but presumably they had IL[12] receptor deficiency, which we were unable to detect in our laboratory. In some patients this marker should be checked after preparation of more laboratory facilities

Shwachman syndrome. Report of three cases

Shwachman syndrome, next to cystic fibrosis, is the second cause of congenital exocrine pancreatic insufficiency in children. It appears as steatorrhea, recurrent infections and hematologic abnormalities such as neutropenia, skeletal dysplasia and short stature. In this study, we reviewed 3 patients' histories. All of them showed cellular chemotaxic defect. One of them had been affected by nephrotic syndrome and finally succumbed to septic shock. This patient suffered from skeletal disorders

Expression of hepatitis B surface antigen in saccharomyces cerevisiae

The genome of HBV virus of serotype ayw cloned in pBR322 and expression shuttle vector pYES2 were used for construction of the HBsAg chimeric genes and their expression in Saccharomyces cerevisiae. Two recombinant plasmids were constructed. One of them contained the coding sequences for the major polypeptide of surface antigen. Another construct carried the major polypeptide with the pre-S2 antigenic determinant. These vectors were transferred into the yeast. Only pDF1 which contained the HBsAg gene was expressed. Some peculiar features of recombinant plasmid construction and expression of the HBsAg gene are discussed

study of leukocyte adhesion disorder [lad]

Recruitment of leukocytes to an inflammatory site includes several steps: rolling, sticking, flattening or tethering and finally diapedesis and migration to the extravascular environment. Leukocyte settlement at any inflammatory site is dependent on adhesion molecules which are selection, integrins, proteins of the immunoglobulin supergene family, and cadherins. In this study we present four cases of leukocyte adhesion disorder [LAD] type 1 which lack LFA-1, Mac-1, P150/95 [CD11a,b,c] and CD18. All of them had recurrent pyogenic infections, leukocytosis and chemotactic defect. Three of these cases had delayed separation of umbilical cord

Isolation, subtype determination, cloning and expression of HB s Ag gene from an Iranian carrier in saccharomyces cerevisiae

The Hepatitis B Surface antigen [HBsAg] gene was isolated from an Iranian HBsAg positive carrier by PCR. The gene was cloned in pUC19 for sequencing and pYES2 for expression in Saccharomyces cerevisiae, which pNFl and pDF3 constructs were made respectively. The sequencing data showed that the isolated HBsAg gene shared more than 90% homology with the ayw subtype. The pDF3 was transferred into the yeast strain and expression of the HBsAg was induced. Yeast-derived HBsAg was purified by immunoaffinity chromatography and checked by ELISA, Western blot analysis and confirmatory ELISA. The immunogenicity test in mice showed that injection of the yeast-derived recombinant HBsAg could induce the immunologic response against the antigen. 320 fold dilution of the mouse serum was positive for anti-HBsAg. The amount of HBsAg expressed in yeast constitute 1% of the total cell soluble protein

Evaluation of serum opsonic capacity against Bordetella pertussis by chemiluminescence

Serum opsonic capacity against B, pertussis was studied by using quantitative chemiluminescence [CL], a method known to have several advantages over conventional methods in evaluating opsonization and phagocytosis. Sera from unvaccinated infants was shown not to contain opsonins against B. pertussis and in unvaccinated infants suffering from whooping cough, no opsonins were detected. In adults fully vaccinated during childhood, antibody liters decreased with time. Therefore, antibody transfer to infants is negligible. The CL assay is simple, rapid, and reproducible, offering new possibilities to evaluate humoral immune mechanisms and phagocytosis in whooping cough

Evaluation of serum IgG subclass levels in asthmatic and atopic children with recurrent infection

Serum IgG subclass levels were measured using an indirect immunoenzymatic assay [ELISA] with monoclonal antibodies in 16 children with asthma and 13 children with atopy who had mostly recurrent infections. Seven of the asthmatic children had marked low or low normal levels of IgG[4], six had marked low or low normal levels of IgG[3], two had marked low normal levels of both IgG[2] and IgG[3] and one had low levels of IgG[2], IgG[3] and IgG[4]. All these patients suffered from recurrent sinopulmonary infections- There were low percentages of IgG[3] and IgG[4] defects [about 15%] in the atopic patients, while a significant increase in the serum IgG[4] levels were observed [six patient out of 13 patients, 46.2%]

case report of common variable immunodeficiency [CVID] with leukocytes adhesion disorders [LADS]

A three year old boy with delayed separation of the umbilical cord, recurrent pyogenic infections and common variable immunodeficiency is presented. The immunoglobulin measurement was severely abnormal. To the best of our knowledge, the existence of leukocyte adhesion disorder together with common variable immunodeficiency [CVID] in a patient has not been reported prior to this study

Drug resistance in diarrhogenic escherichia coli [ETEC,EPEC]

A total of 429 enteropathogenic and enterotoxigenic E. coli isolated from diarrhoeal cases during 1986 and 1989 was tested and compared for resistance against 10 commonly used antimicrobial agents by disk diffusion method. The highest resistance in both years and among all strains was against ampicillin and the least against gentamicin. Multiresistance was common among the isolates with A/C/T/SXT and A/C/T/SXT/S comprising the most prevalent pattern. This study, which is also the first report of antimicrobial drug resistance among ETEC, underlines the need for regular surveillance and its bearing on drug policy is discussed

study of opsonization activity of the phagocytosis process in meningitis, sepsis and recurrent infection in children

In this survey, opsonization activity of sera in 110 patients with meningitis, sepsis and recurrent infection; 12 patients after healing; and 50 normal adults has been studied. Opsonization activity was measured by an assay depending on electronic particle counting to measure yeast uptake [alternative pathway activator] which was modified in our laboratory. Three classes of immunoglobulins [lgG, 1gM, igA] and C3 levels were also studied depending on single radial immunodiffusion by using Behring kit in our patients. 54 pathological sera with defective opsonization showed low C3 levels, two with lower than normal opsonization activity showed low C3 levels, 11 defective opsonization cases showed normal C3 levels, 3 of which were related to diabetic patients. 22 instances of decreased lgG, 14 cases of decreased 1gM and 18 cases of decreased lgA were observed in 110 patients. The opsonization activity, C3 and immunoglobulin levels in 12 patients have become normal after healing

Neutrophil chemotactic defect in 60 patients with rickets

Neutrophil mobility in 60 patients with vitamin D-deficient rickets is studied, and the patients are followed after therapy by 600,000 units vitamin D. 28 of 60 patients [46%], had low chemotaxis for neutrophils before vitamin D injection [migration], and 36 of 60 [60%] had low random mobility. The results showed a significant improvement after vitamin D therapy; furthermore, the level of calcium and phosphorus reached normal levels after treatment of the patients as well. We have previously studied neutrophil mobility in 10 patients with rickets. This report serves to complement our previous preliminary report

Defective neutrophil mobility in ten patients with vitamin D-deficient rickets

We studied the immune system, especially the chemotactic activity of neutrophils, in ten patients with vitamin D-deficient rickets and compared the results with ten healthy controls of matched age. Among all immune system factors, the chemotactic studies persistantly showed remarkable deficiency in leukocyte mobility, both random motion and migration. When compared with controls, the findings were significant. It was postulated that the increased susceptibility of the patients with vitamin D-deficient rickets to infection is mainly due to defective neutrophil mobility