Role of interleukin 4 (IL4) and interleukin 6 (IL6) in the pathogenesis and prognosis of childhood primary immune thrombocytopenia.

Immune thrombocytopenia (ITP) is an autoimmune disease characterized by the breakdown of immune tolerance. Impairment of the cellular immunity is primarily evaluated by the levels of the cytokines which can help in predicting the course of ITP. We aimed to assess the levels of IL4 and IL6 in children with ITP and evaluate their role in the pathogenesis and prognosis of this disease. A prospective cohort study was carried on 60 children (15 patients with newly diagnosed ITP, 15 patients with persistent ITP, 15 patients with chronic ITP and 15 healthy children as a control group). Serum IL-4 and serum IL-6 were measured using Human IL-4 and IL-6 ELISA kit in patients and controls. Patients with newly diagnosed and persistent ITP had significantly higher levels of IL4 and IL6 compared to patients with chronic ITP and healthy controls (p < 0.001). The mean serum level of IL4 was 762.0, 741.0, 364.6 and 436.8 pg/ml, and the mean serum level of IL6 was 178.5, 164.4, 57.9 and 88.4 pg/ml for patients with newly diagnosed, persistent, chronic ITP and healthy controls respectively. Serum IL-4 was significantly higher in patients who achieved remission than those who did not improve on first line therapy. CONCLUSION: Serum IL-4 and IL-6 may have a role in the pathogenesis of primary ITP. IL-4 seems to be a good predictor to treatment response. WHAT IS KNOWN: • There is a delicate balance of specific cytokine levels in immune thrombocytopenia, which has an important role in the immune system and is known to be deregulated in autoimmune diseases. changes in IL-4 and IL-6 might be involved in the pathogenesis of newly diagnosed ITP in both paediatric and adult patients. • We conducted this research study to measure the serum level of IL-4 and IL-6, in newly diagnosed, persistent and chronic ITP patients and study their relation to disease pathogenesis as well as patient's outcome. WHAT IS NEW: • We found that IL4 seems to be a good predictor to treatment response and it was a very interesting observation in our study, and to the best of our knowledge, there is no published data about this finding.

Tissue factor expression predicts outcome in children with neuroblastoma: A retrospective study.

Previous studies have revealed that the processes of tumor angiogenesis, metastasis and invasiveness are highly dependent on components of the blood coagulation cascade. Tissue factor (TF) is one of the key proteins in coagulation. Cumulative evidence suggested that in addition to its role in coagulation, TF regulates intracellular signaling pathways that serve an important role in angiogenesis, tumor development and metastasis. In the present study, TF expression in neuroblastoma as well as its association with tumor stage, pathology and outcome were assessed. A total of 40 formalin-fixed paraffin-embedded tissues were evaluated for TF expression by immunohistochemical analysis. Results revealed that TF expression was positive in 75% of the analyzed tumor tissues. No significant association between TF expression and sex, age, tumor stage or disease pathology was observed. MYCN proto-oncogene bHLH transcription factor (MYCN) was upregulated in 45% (n=18) of the study cases. Positive TF expression was observed in 94.4% of patients (n=17) with upregulated MYCN, while 59% of patients (n=13) with normal MYCN showed positive TF expression (P<0.05). TF expression was a significant outcome predictor for patients; 18/30 patients (60%) with positive TF expression succumbed to the disease during the study period. In conclusion, TF may be a promising prognosis indicator for neuroblastoma. Future studies to determine how TF affects the progression and outcome of neuroblastoma, as well as to investigate its potential role as a therapeutic target, are required.

Microbial etiology of community acquired pneumonia among infants and children admitted to zagazig university pediatric hospital

Background: While recognizing the etiology of community-acquired pneumonia is necessary for formulating local antimicrobial guidelines, limited data is published about this etiology in Egyptian pediatric patients. The aim of this study is to elucidate the common bacterial pathogens causing CAP among immunocomptent infants and preschool children admitted to Zagazig university Pediatric hospital. Methods: 48 infant and preschool children admitted to pediatric hospital of Zagazig university and presented with signs of pneumonia according to WHO. Etiological agents were identified using conventional bacteriological identification methods and Ig M antibodies detection against common a typical bacteria and respiratory viruses. Results: Staph. Aureus 35.4% is the most common pathogen detected in sputum regardless the age group .In blood culture results negative results in most cases of pneumonia 62.5%. Staph. Aureus is the most common pathogen detected in blood culture in positive cases regardless the age group 18.7% , E Coli 6.25%, Klebsiella 2.08.In Serology results significant difference according to age in viral pneumonia occurrence and non significant difference in bacterial pneumonia based on serological findings with higher diagnosis of Legionella pneumophila 33.33% as a causative organism. Conclusion: This study provides preliminary data regarding the spectrum and frequency of microorganisms causing CAP in infants and preschool children

Impact of iron deficiency anemia on the function of the immune system in children.

The importance of iron deficiency as a public health problem is based ultimately on the seriousness of its consequences on health. The most extensively investigated consequences of iron deficiency involve work performance and immune function. The significance of the effects on work performance is generally accepted. In contrast, data on the influence of iron deficiency on immune function are often perceived as being confusing and contradictory.We aimed to evaluate the effect of iron deficiency anemia on humoral, cellular, nonspecific immunity, and also the effect on the cytokines that are the key factors of many immunologic steps.Forty children with iron deficiency anemia and 20 age and sex-matched healthy children were included. All children were subjected to full medical history, thorough clinical examination, complete blood count, iron indices (serum iron, serum total iron-binding capacity, serum ferritin, and transferrin saturation), immunoglobulin assay (IgA, IgG, and IgM), interleukin (IL)-6 serum level, study of T-lymphocyte subsets, and evaluation of phagocytic function of macrophages and oxidative burst activity of neutrophils.Patients had significantly lower IgG levels, IL-6, phagocytic activity, and oxidative burst of neutrophils than controls, although there was no significant difference between patients and controls with regard to other immunoglobulins and CD4/CD8 ratio. There was significantly positive correlation between serum iron and IL-6 serum level.We concluded that humoral, nonspecific immunity (phagocytic activity and oxidative burst), and the IL-6 are influenced in patients with iron deficiency anemia. Study of these abnormalities after correction of iron deficiency is strongly needed.

Significance of CD34/CD123 expression in detection of minimal residual disease in B-ACUTE lymphoblastic leukemia in children.

BACKGROUND: MRD is seen as the major cause of disease relapse. So, it gives important feedback about conventional treatment success and helps in selecting therapeutic alternatives. We aimed to compare the expression of CD34/CD123 on normal B-cell precursors in bone marrow ("hematogones") and on leukemic blasts in B-acute lymphoblastic leukemias (B-ALL) pediatric cases by flowcytometric analysis. Our study conducted on 20 children as a control and 30 B-ALL children cases at diagnosis and after 28days of induction therapy. We found that the less mature hematogones (dim CD45+) that express CD34 lack CD123 expression, whereas the more mature hematogones (moderate CD45+) lack CD34 but always express CD123. In contrast with this discordant pattern of CD34 and CD123 expression in hematogones, blasts in 24 of 30 cases (80%) of B-ALL showed concordant expression pattern of the 2 antigens: 63% (19 of 30) cases expressed both antigens, whereas 17% (5 of 30) expressed neither. Our study concluded that these distinct patterns of CD34/CD123 expression on hematogones (discordant) and B-ALL blasts (concordant) are useful in differentiating small populations of residual blasts from hematogones after induction therapy to detect MRD.

Serum trace elements in obese Egyptian children: a case-control study.

BACKGROUND: To date, only a few studies on child obesity concerned Trace Elements (TE). TE is involved in the pathogenesis of obesity and obesity related diseases. We tried to assess trace elements status [zinc (Zn), copper (Cu), selenium (Se), iron (Fe), and chromium (Cr)] in obese Egyptian children and their relationships with serum leptin and metabolic risk factors of obesity. METHODS: This was a case-control study performed with 80 obese children (BMI ≥ 95thcentile for age and gender) and 80 healthy non-obese children with comparable age and gender as the control group. For all subjects, serum Zn, Cu, Se, Fe, ferritin and Cr as well as biochemical parameters including lipid profile, serum glucose and homeostasis model assessment of insulin resistance (HOMA-IR) were assessed. Levels of serum leptin were measured by (enzyme-linked immunosorbent assay [ELISA] method), and serum insulin was measured by an electrochemiluminesce immunoassay. RESULTS: Compared to the control group, serum Zn, Se, and Fe levels were significantly lower (all P < 0.01) and serum Cu level was significantly higher (P < 0.01) in the obese children. Meanwhile, no significant differences were observed in serum ferritin or Cr levels (P > 0.05). A significant negative correlation was found between serum leptin and zinc levels in the obese children (r = -0.746; P < 0.01). Further, serum Zn showed significant negative correlations with total cholesterol TC levels (P < 0.05) and were positively correlated with high density lipoprotein- cholesterol HDL-C levels (P < 0.01) in the obese children. In addition, serum Se levels showed significant positive correlations with HOMA-IR values in the obese children (P < 0.01). CONCLUSION: The obese children may be at a greater risk of developing imbalance (mainly deficiency) of trace elements which may be playing an important role in the pathogenesis of obesity and related metabolic risk factors.

Iron deficiency anemia as a risk factor for cerebrovascular events in early childhood: a case-control study.

In recent years, iron-deficiency anemia (IDA) has been suggested to have an association with childhood-onset ischemic stroke in otherwise healthy children, but few cases have proven it thus far. In this study, we aimed to investigate whether iron-deficiency anemia is a risk factor for cerebrovascular events and childhood-onset ischemic stroke in previously healthy children. This was a case-control study that included 21 stroke cases with patients who had previously been generally healthy, and matched with age and gender of 100 healthy control subjects. Patients were included if a diagnosis of definite stroke had been made and other known etiologies of childhood onset stroke were excluded. For all subjects, iron parameters including serum iron, ferritin, transferrin, total iron binding capacity, and transferrin saturation were assessed. We screened all case patients for prothrombotic factors including level of hemoglobin S, protein C, protein S, antithrombin III, lupus anticoagulant, factor V Leiden, and prothrombin gene mutation (G20210A). Brain magnetic resonance images (MRI), magnetic resonance angiography (MRA), and magnetic resonance venography (MRV) were performed to all case patients. All case patients have normal results regarding functional, immunological, and molecular assay for prothrombotic factors screening. Our results showed that IDA was disclosed in 57.1 % of stroke cases with no identified cause, as compared to 26 % of controls. Our study suggest that previously healthy children who developed stroke are 3.8 times more likely to have IDA than healthy children, who do not develop stroke (OR, 3.8; 95 % CI:1.3-11.2 P = 0.005). In addition, there was significant interaction between IDA and thrombocytosis among studied cases (OR, 10.5; 95 % CI, 1.0-152 P = 0.02). There were nonsignificant differences between stroke patients with IDA and those with normal iron parameters regarding stroke subtype (P > 0.05). Public health messages on the importance of early detection of iron-deficiency anemia in young children, especially in our developing countries so that it can be treated before a life-threatening complication like stroke develops.

Vitamin D status in diabetic Egyptian children and adolescents: a case-control study.

BACKGROUND: Recently, studies suggesting that vitamin D deficiency correlates with the severity and frequency of Type 1 (insulin-dependent) diabetes mellitus (T1DM) and that vitamin D supplementation reduces the risk of developing T1DM have been reported. OBJECTIVE: In this study, we aimed to assess vitamin D status in Egyptian children and adolescents with T1DM. METHODS: This was a case-control study including 80 T1DM diagnosed cases aged 6 to 16 years and 40 healthy children with comparable age and gender as the control group. For all subjects, serum 25 (OH) D levels were measured by ELISA, Serum parathyroid hormone (PTH) and serum insulin were measured by an electrochemiluminesce immunoassay. Serum glucose, Glycosylated hemoglobin (HbA1c) levels and homeostasis model assessment of insulin resistance (HOMA-IR) were also assessed. RESULTS: Compared to the control group, serum vitamin D levels were not significantly lower in diabetic subjects (24.7 ± 5.6 vs 26.5 ± 4.8 ng/ml; P > 0.05). Among diabetic cases 44(55%) were vitamin D deficient; meanwhile 36(45%) cases had normal vitamin D level (P < 0.01). In addition, 26(32.5%) diabetic cases had 2ry hyperparathyroidism and 54(67.5%) cases had normal parathyroid hormone level; meanwhile, none of the control group had 2ry hyperparathyroidism (P < 0.01). Furthermore, we found a significant difference between vitamin D deficient diabetic cases and those with normal vitamin D level as regards HOMA-IR and diabetes duration (P < 0.01). CONCLUSION: Public health message on the importance of vitamin D status; especially in diabetic children and adolescents, should be disseminated to the public.

Serum hepcidin levels in Helicobacter pylori-infected children with iron-deficiency anemia: a case-control study.

Recently, hepcidin, an antimicrobial-like peptide hormone, has evolved as the master regulator of systemic iron homeostasis. Hepcidin integrates signals from diverse physiological inputs, forming a key connection between iron trafficking and response to infection. In this study, we aimed to investigate whether Helicobacter pylori infection modulates serum hepcidin level and response to oral iron therapy in children with iron-deficiency anemia. This was a case-control study including 60 children with iron-deficiency anemia (IDA; 30 H. pylori infected and 30 H. pylori noninfected) and 30 healthy children with comparable age and gender as the control group. Iron parameters including serum iron, ferritin, transferrin, total iron binding capacity, and transferrin saturation and serum hepcidin levels were assessed initially and after 3 months of oral iron therapy for IDA. Compared to the control group, serum hepcidin was significantly lower in H. pylori-noninfected children with IDA (P < 0.01) and significantly higher in H. pylori-infected children with IDA (P < 0.01). Hepcidin increased significantly in noninfected children with IDA after 3 months of oral iron therapy (P < 0.01). On the other hand, H. pylori-infected children showed nonsignificant change in hepcidin level after oral iron therapy (P > 0.05). Although hepcidin showed significant positive correlations with serum ferritin, hemoglobin (Hb), iron, and transferrin saturation in noninfected children with IDA (P < 0.01), it showed significant negative correlations with serum ferritin, Hb, iron, and transferrin saturation in H. pylori-infected children with IDA (P < 0.05). H. pylori infection upregulates serum hepcidin levels and was associated with diminished response to oral iron therapy in children with iron-deficiency anemia.

Serum NGAL in Critically Ill Children in ICU from a Single Center in Egypt.

Introduction. The mortality and morbidity associated with acute kidney injury (AKI), unfortunately, remain unacceptably high. We aimed to detect the extent of serum neutrophil gelatinase-associated lipocalin (NGAL) to early detect AKI in critically ill children. Subjects and Methods. This is a case control study. It included 75 subjects that include 15 as controls and 60 critically ill children. Patients were further subdivided according to RIFLE criteria into two other categories: patients who developed AKI and patients who did not develop AKI. Serum NGAL assayed on admission and after 3 days. Results. There was significant increase in the level of NGAL among patients group when compared with control group. Also, 21.7% of children admitted to PICU developed AKI from which 8.3% needed dialysis. The receiver operating characteristic curve of NGAL at day 0 revealed AUC of 0.63 with 95% CI of 0.50-0.77. At a cutoff value of 89.5 ng/mL, the sensitivity of NGAL was 84.6%, while specifcity was 59.6%, positive predictive value was 36.7%, negative predictive value was 68.4%, and accuracy was 93.3% in diagnosis of AKI. Conclusion. We found that NGAL acts as a sensitive marker rather than a specific one for AKI. At the same time, it presents as a negative predictive value more valuable than being a positive predictive value in detecting AKI.

Biomarkers and early detection of late onset anthracycline-induced cardiotoxicity in children.

BACKGROUND: The main strategy for minimizing anthracycline cardiotoxicity is early detection of high-risk patients. AIM OF THE STUDY: To investigate the role of cardiac biomarkers; cardiac troponin T (cTnT) and N-terminal probrain natriuretic peptide (NT-pro-BNP), and tissue Doppler imaging (TDI), as early predictors of chronic cardiotoxicity in survivors of acute leukemia. PATIENTS AND METHODS: We carried a retrospective study on 50 asymptomatic survivors of acute leukemia who received anthracycline in their treatment protocols. All patients underwent blood sampling to determine the levels of NT-pro-BNP and cTnT along with conventional echocardiography and TDI. RESULTS: None had abnormal cTnT levels. About 20% had abnormal NT-pro-BNP levels. Diastolic dysfunction of the left ventricle was the most significant in conventional echocardiography. TDI was superior as it detected myocardial affection in 10% more than echo. TDI demonstrated global myocardial damage with significant aberrations in peak myocardial velocities and ratios. CONCLUSIONS: NT-pro-BNP can be used as a sensitive cardiac biomarker in monitoring of anthracycline-induced cardiotoxicity. Follow up is essential to validate the role of NT-pro-BNP as an early marker for late onset anthracycline-induced cardiotoxicity. Tissue Doppler is marvelous as it could detect early cardiac dysfunction even in those with normal study by conventional echocardiography.

Prognostic significance of survivin in pediatric acute lymphoblastic leukemia.

Impaired apoptosis is mediated by members of the inhibitor of apoptosis proteins (IAP) family such as survivin. Survivin was described in number of different tumors and found to correlate with poor prognosis in a variety of cancers including hematologic malignancies. The aim of this study was to determine survivin in pediatric ALL and compare it with clinical and hematological findings, response to therapy and outcome. Flowcytometry was used for detection of intracellular survivin and determine its mean fluorescence intensity (MFI) in bone marrow mononuclear cells. Patients were followed up for 28 months after induction therapy. Survivin was detected in 63.3% of the patients BM. In spite of no association of survivin levels with established risk factors (P > 0.05) except with high WBC, there was significant higher level of survivin expression in high risk group patients when patients were stratified into high and standard risk groups. According to response to induction therapy, there was no significant difference, in survivin level between patients who achieved CR, RD and ED. However, patients suffering relapse of the disease, had a significant higher basal level of survivin than patients still in remission. Over expression of survivin is a candidate parameter to determine poor prognosis in ALL patients and it may serve to refine treatment stratification with intensification of therapy in those patients prone to relapse.

Impact of CD4+CD25high regulatory T-cells and FoxP3 expression in the peripheral blood of patients with systemic lupus erythematosus.

Systemic lupus erythematosus (SLE) is a systemic autoimmune disease characterized by various immunological abnormalities. Regulatory T cells (Tregs) CD4+CD25+ play an important role in maintaining tolerance to self-antigens controlling occurrence of autoimmune diseases. It has been shown that the transcription factor forkhead box P3 (FoxP3) is specifically expressed on CD4+CD25+T cells. FoxP3 has been described as the master control gene for the development and function of Tregs. A decrease in the number of CD4+CD25highFoxP3+ regulatory T cells can play a key role in the loss of tolerance to self antigens. The study was designed to assess expression levels of FoxP3 in peripheral CD4+CD25+ regulatory T cells in patients with SLE and to evaluate the level of some cytokines that are implicated in the extent of the disease activity. The study was carried out on 30 SLE patients, they were 27 females and 3 males, 10 age and sex matched healthy volunteers were studied as a control group. They were divided into two groups: group I: had active disease (12 patients) and group II: had inactive disease (18 patients) according to Systemic Lupus Erythematosus Disease Activity Index. All individual were subjected to CBC, ESR, s.creatinine, RF, CRP, C3, ANA, anti ds-DNA and flowcytometeric assay of CD4+CD25+ (Tregs) and FoxP3 for patients and controls. Quantitative determination of serum interleukin 10 (IL-10) and transforming growth factor-beta1 (TGF-beta1) concentrations in serum samples by ELISA technique. The results revealed a significant decrease of CD4+CD25high cells in peripheral blood in active lupus patients when compared with patients with inactive lupus and those in healthy controls. Intriguingly, the percentage level of FoxP3 on CD4+CD25high cells was significantly decreased in SLE patients with active disease (2.9 +/- 1.05) when compared with those with inactive SLE (3.5 +/- 0.8) and control groups (4.7 +/- 1.2) (P < 0.05). As regard cytokines levels; the level of IL-10 was significantly increased in patients with active and inactive disease (158.8 +/- 50.8, 82.8 +/- 14.08 respectively) when compared with the control group (P < 0.001). While, the level of TGF-beta1 was significantly decreased in patients with active and inactive disease (22.5 +/- 7.03, 29.07 +/- 10.14 respectively), when compared with the control group (P < 0.05). Our data revealed impaired production of Tregs in SLE patients, which may play a reciprocal role with some cytokines to affect the activity of the disease. Tregs cells should be the target to determine the clinical effectiveness of novel therapy to modulate Tregs in vivo besides the conventional treatments.