Stromal Derived Factor-1 Gene Polymorphism in Pediatric Immune Thrombocytopenia.

Immune thrombocytopenia (ITP) is a multifactorial disease in which both environmental and genetic factors have been implicated. The study aimed to investigate a possible association of single nucleotide polymorphisms (SNPs rs266085 and rs2839693) in the stromal derived factor-1 (SDF-1) gene and its association to ITP and effect on ITP severity and response to treatment. Genomic DNA was extracted from peripheral blood and polymorphism in SDF-1 gene rs266085 and rs2839693 was analyzed using PCR-restriction fragment length polymorphism technique in DNA extracted from 60 children with ITP together with 90 healthy controls. On analysis of SDF-1 rs266085 polymorphism, there was a high frequency of CC genotype in cases than controls and that difference was significant at codominant, overdominant, and dominant models (P<0.05). Furthermore, carriers of the CC genotype were more susceptible to severe ITP at onset, steroid dependency, and chronicity than carriers of other genotypes (P<0.05). Otherwise, no significant differences between ITP patients and controls as regard SDF-1 rs2839693 genotypes and alleles, and we did not find a relation between this polymorphism and ITP severity, steroid dependency, or duration. SDF-1 gene rs266085 SNP C allele is associated with susceptibility to develop ITP as well as increases the risk for severe ITP at onset, chronic ITP and steroid dependency.

IL-8 gene expression and bronchial asthma phenotypes in children.

Asthma is a common chronic illness among school children, where different cytokines, including IL-8 play a role in its pathogenesis. IL-8 induces chemotaxis and migration of immune cells, especially neutrophils to the site of inflammation. IL-8 level was significantly increased in sputum of severely asthmatic patients, but can it be linked to some asthma phenotypes. Our aim of the study was to detect the IL 8 gene expression in different asthma phenotypes and to determine its relation to asthma severity. This case control study included 320 subjects (160 asthmatic and 160 matched controls) aged from 5 to 16 years old in Beni-Suef governorate. IL-8 gene expression was assessed by a real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) and studied regarding its level in cases versus controls and its relations to severity, phenotype and other laboratory parameters. IL-8 gene expression was statistically higher in asthmatic cases (P<0.001) and was significantly correlated to the phenotype (presence of other allergy as urticaria and drug eruption) and degree of asthma symptoms (r=0.869, P<0.001), FEV1(r=0.757, P<0.001) and serum IgE level (r=0.789, P<0.001). IL-8 gene expression level is increased with the degree of severity in asthmatic children and can be looked for in certain asthma phenotypes especially in presence of other atopic manifestation.

Expression of Serum MicroRNAs 221, 222, 15a and Level of VEGF-A in Children with Bronchial Asthma.

Bronchial asthma (BA) remains the most common chronic respiratory disease in children and is characterized by reversible airway obstruction and airway hyperresponsiveness. MicroRNAs (miRNAs) are a class of small, highly conserved non-coding RNA molecules that regulate gene expression at the posttranscriptional level. MiRNAs can either augment or minimize the allergic inflammation in asthma. It has been noted that a group of miRNAs may affect the development of asthma and related inflammatory response. The vascular endothelial growth factor (VEGF) is produced by different types of cells and has a major role in both, physiological and pathological angiogenesis. Our aim was to study serum expression of three microRNAs; 221, 222,15a and Vascular Endothelial Growth Factor A (VEGF-A) levels in children with bronchial asthma. The study included 30 children with BA and a control group of 20 apparently healthy, age and sex matched, children. Quantitative reverse transcription (q RT) polymerase chain reaction (PCR) was performed to examine the expression levels of miR-221, 222 and 15a in patients' sera. Levels of serum VEGF-A were quantified utilizing an ELISA technique. Serum levels of both miRNA-221 and miRNA-222 were not significantly different between pediatric asthmatic patients versus controls (P=0.76 and 0.52, respectively), but showed increase with the disease severity (persistent versus intermittent) (P= 0.09, 0.07, respectively). Serum miRNA-15a expression was significantly down-regulated in asthmatic patients versus the control group (P=0.03). miRNA-15a expression did not differ among various grades of BA (P=0.33) and was not correlated with serum levels of VEGF-A (P=0.56). The level of VEGF-A was significantly increased in serum of pediatrics with bronchial asthma in comparison to the control group (P=0.026). In conclusion, miRNA-15a and VEGF-A may have a role in BA pathogenesis, while miRNA-221 and 222 may reflect BA disease severity.

Skin Disorders in Egyptian Children with ß-Thalassemia Major.

Background: Thalassemia major is one of the common genetic disorders in Egypt. Skin disorders are usually neglected and frequently underdiagnosed among these patients. Objective: This work aimed to study the frequency and pattern of skin manifestations in Egyptian children with ß-thalassemia. Methods: Fifty-four ß-thalassemia major patients being followed at the Hematology Clinic of Beni-Suef University Hospital were selected to participate in this study. All patients underwent detailed history evaluation, clinical examination and laboratory investigations, including complete blood count and serum ferritin. All patients were examined by a dermatologist to record any skin disorders. Results: Pruritus (37%), scars (33.3%), hyperpigmentation (31.5%) and xerosis (22.2%) were the most common findings. We found significant relations between serum ferritin and the occurrence of scars, hyperpigmentation, xerosis and ephelides (p <0.05). Also, significant associations between use of deferoxamine and scars (p=0.004), hyperpigmentation (p=0.004), xerosis (p=0.03) and ephelides (p=0.042) were found. Conclusion: Skin disorders are frequent in Egyptian children with thalassemia major. Careful skin examination is required to provide early diagnosis.

Evaluation of aldosterone excretion in very low birth weight infants.

Data about aldosterone production and excretion in the neonatal period are still few and controversial. Our objectives are to assess urinary aldosterone excretion (UAE) in very low birth weight (VLBW) infants and to identify clinical and biochemical variables that may influence this excretion. Thirty VLBW infants (14 males and 16 females), their gestational age <32 weeks and body weight <1500 g, were included in the study. Demographic and clinical data were recorded, within the first 72 h of life and urine and blood samples were collected for the measurement of urinary aldosterone and serum potassium, sodium, and chloride. The mean UAE value was 0.176 ± 0.05 µg/24 h and the mean absolute UAE was 1906 ± 271 pg/mL. There was a statistically significant positive correlation between UAE and gestational age and birth weight; also, infants with respiratory distress syndrome had higher urinary aldosterone levels than infants without respiratory distress. Only plasma sodium was a significant independent factor that negatively influenced UAE on linear regression analysis. The renin-angiotensin-aldosterone system of VLBW infants seems to be able, even immediately after birth, to respond to variations of plasma sodium concentrations; measurement of UAE constitutes an interesting method to determine aldosterone production in VLBW infants.

Autoimmune thrombocytopenia: flow cytometric determination of platelet-associated CD154/CD40L and CD40 on peripheral blood T and B lymphocytes.

BACKGROUND AND OBJECTIVES: The CD40-CD40L system has pleiotropic effects in a variety of cells and biological processes including the immune response. Within the immune system, these molecules represent a critical link between its humoral and cellular arms. Immune or idiopathic thrombocytopenic purpura (ITP) is an autoimmune disorder characterized by antibody-induced platelet destruction and clearance due to anti-platelet autoantibodies, which bind to circulating platelets resulting in their destruction by the reticuloendothelial system. Despite its clinical importance, the diagnosis of ITP is one of exclusion, thus, inevitably associated with potential difficulties. CD40 is a cell surface receptor that belongs to the tumor necrosis factor-receptor (TNF-R) family, and that was first identified and functionally characterized on B lymphocytes. CD40-ligand (CD40L/CD154), a member of the TNF superfamily, is a cell membrane molecule expressed on activated CD4 + T lymphocytes and is essential for the T cell-dependent activation of B lymphocytes. Therefore it is now thought that CD40-CD40L interactions play a more important role in ITP immune regulation. DESIGN AND METHODS: The expressions of CD154 and CD40 on peripheral blood (PB) T and B lymphocytes, respectively, were measured using flow cytometry (FCM). An antigen-specific assay for platelet-associated CD154 (CD40L) on CD4 + T lymphocytes and for CD40 on CD19 + B lymphocytes was tested in 30 pediatric patients with acute ITP, 30 adult patients with chronic ITP, and in 20 age- and sex-matched healthy controls. RESULTS: The expression of CD4 + CD154+ and CD4 + CD154+/CD4+ on PB T lymphocytes, and CD19 + CD40+ and CD19 + CD40+/CD19+ on PB B lymphocytes were significantly higher in acute and chronic ITP patients compared to controls, and in acute patients compared to chronics (p < 0.001). CONCLUSIONS: CD40-CD40L interaction plays an important role in the pathology of certain autoimmune diseases. ITP is an autoimmune disease characterized by increased platelet destruction caused by anti-platelet autoantibodies, which mainly target a platelet surface antigen. It is speculated that platelet-associated CD154 is competent to induce the CD40-dependent proliferation of B lymphocytes. Therefore, platelet-associated CD154 expression is increased in ITP patients and is able to drive the activation of autoreactive B lymphocytes in this disease. These findings are particularly useful for clarifying the pathogenic process in ITP patients and for developing a therapeutic approach that blocks pathogenic anti-platelet antibody production. Blockade of the CD40/CD154 signal is a potential immunomodulatory strategy for T cell-mediated diseases, and many findings suggest that CD40/CD154 blockade therapy is potentially effective for ITP through selective suppression of autoreactive T and B lymphocytes to platelet antigens.

Safety and efficacy of rifampicin in children with cholestatic pruritus.

OBJECTIVE: The present study aimed at verifying the safety and efficacy of rifampicin in ameliorating pruritus in cholestatic children. METHODS: Twenty-three Egyptian children (14 boys and 9 girls), suffering from intractable pruritus of cholestasis, were included. Rifampicin was started at a dose of 10 mg/Kg/day in two divided doses and increased gradually to a maximum of 20 mg/Kg/day if there was no response. Liver function tests were followed up weekly. RESULTS: Seventeen patients (74%) showed improvement of pruritus with rifampicin. None of the patients showed any deterioration in liver functions. CONCLUSION: Rifampicin in a dose of 10-20 mg/Kg/day is safe and effective in ameliorating uncontrollable pruritus in children with persistent cholestasis.

Safety and efficacy of hepatitis A vaccine in children with chronic liver disease.

AIM: To study the safety and efficacy of hepatitis A vaccine (HAV) in children with chronic liver disease of various etiologies. METHODS: Eleven children with chronic liver disease and thirteen age- and sex-matched controls negative for HAV antibodies were vaccinated against hepatitis A after they gave their informed consent. Children with uncontrolled coagulopathy or signs of hepatic decompensation were excluded. The vaccine (Havrix: 720 ELISA units in 0.5 mL, from GlaxoSmithKline Biologicals) was given intramuscularly in the deltoid in 2 doses 6 mo apart. Children were tested for HAV antibodies one and six months after the 1st dose and one month after the 2nd dose. Total serum bilirubin, alanine aminotransferase (ALT), and aspartate aminotransferase (AST) were determined immediately before and after one month of the 1st dose of the vaccine. RESULTS: Only 7 out of the 11 patients were positive for HAV antibodies after the 1st dose of the vaccine, as compared to 100% of the controls. One month after the 2nd dose, all patients tested were positive for HAV antibodies. No deterioration in liver functions of patients was noted after vaccination. No adverse events, immediate or late, were reported by the mothers after each dose of the vaccine. CONCLUSION: Hepatitis A vaccine is both safe and effective in this small studied group of children with chronic liver disease. Given the high seroconversion rate, post-vaccination testing for HAV antibodies is not needed.