Iman Ragab

Evaluation of the immature platelet fraction in the diagnosis and prognosis of childhood immune thrombocytopenia

Abstract Rapid assessment of platelet production would distinguish between thrombocytopenia due to decreased platelet production or increased peripheral platelet destruction. We evaluated the value of immature platelet fraction (IPF) in differentiating immune thrombocytopenia (ITP) from thrombocytopenia secondary to bone marrow failure and its potential use as a prognostic marker. Forty-one young patients with ITP were compared with 14 patients with hematological malignancies under chemotherapy, representing a control group with thrombocytopenia due to bone marrow suppression and 30 age- and sex-matched healthy controls. Patients were studied stressing on bleeding manifestations, organomegaly/lymphadenopathy and therapy. Complete blood count including IPF was performed using Sysmex XE-2100. ITP patients were classified into two subgroups: acute ITP with spontaneous resolution within 3 months from diagnosis and chronic ITP that lasted ≥1 year from diagnosis. Median IPF was 11.8% in patients with ITP, 7% in those with hematological malignancy and 3% in the control group (p < 0.001). ITP patients had significantly higher mean platelet volume (MPV), platelet distribution width (PDW), platelet large cell ratio (P-LCR) and IPF compared with patients with malignancy or healthy controls, while plateletcrit (PCT) was significantly lower in ITP patients than other groups (p < 0.001). IPF was increased in patients with chronic ITP compared with acute ITP group (p < 0.001). Patients with active ITP had the highest IPF followed by those in partial remission, while ITP patients in remission had the lowest IPF. IPF was positively correlated to the number of lines of treatment used, MPV, PDW and P-LCR, while negatively correlated to platelet count and PCT among ITP patients (p < 0.001). Multiple regression analysis showed that platelet count and P-LCR were independently related to IPF. ROC curve analysis revealed that the cut-off value of IPF at 9.4% could be diagnostic for ITP patients with a sensitivity of 88% and a specificity of 85.7%. We suggest that IPF may be a rapid and inexpensive automated marker for etiology of thrombocytopenia and can be integrated as a standard parameter to evaluate the thrombopoietic state of the bone marrow. It may be considered as a potential prognostic marker for the development of chronic ITP.

Neurocognitive Outcome and White Matter Anisotropy in Childhood Acute Lymphoblastic Leukemia Survivors Treated with Different Protocols

Neurocognitive outcome affects the quality of life of ALL survivors. This study is aimed to assess the prevalence of neurocognitive dysfunction by psychometric and imaging tools in survivors of childhood ALL, treated with 3 different protocols and the effect of time elapsed since the end of chemotherapy. Sixty-two ALL survivors aged 6–18 years and treated in the period 1997–2007 and 60 healthy age and sex matched controls were subjected to neurocognitive testing using Wechsler Intelligence Scale for Children, Benton visual retention (BVRT) and Trail Making test (TMT), followed by diffusion weighed and diffusion tensor MRI for calculation of fraction anisotropy (FA). Survivors underwent revision of protocol and type of CNS therapy. Three different protocols were used: modified BFM 83, BFM 90, and CCG. Survivors treated with modified CCG protocol showed a significant decrease in all cognitive tests compared to control (p<.05); BFM 90 group had a significant lower IQ and longer TMT compared to both control and BFM 83 group and no significant difference was found in results of cognitive tests between BFM 83 and control group. Frontal FA was lower in CCG treated group compared to control, BFM 90 and BFM 83 groups (p<.05); meanwhile it was significantly lower in BFM 90 and BFM 83 groups compared to control group. We concluded that patients treated with modified CCG protocol showed the worst neurocognitive outcome among three assessed protocols. Frontal lobe FA might be an early marker for predicting the neurotoxicity in childhood ALL survivors.

Pathogenesis and prognosis of neutropenia in infants and children admitted in a university children hospital in Egypt.

This study aimed to assess the prevalence, severity, and etiology of neutropenia in infants and children admitted to a childrens hospital in Egypt. A total of 200 patients with neutropenia were recruited from April 1, 2010 to September 30, 2010. Patients with a known hematological or immunological disease were excluded. Patients were followed till recovery or an underlying cause was uncovered. Viral serological analysis was done for patients with moderate/severe neutropenia, including cytomegalovirus (CMV); Epstein–Barr virus (EBV); hepatitis A, B, and C viruses; and HIV. Antineutrophil cytoplasmic antibody (ANCA) tested by enzyme immunoassay and bone marrow aspirate were done for prolonged neutropenia. The results revealed that neutropenia was mild in 90 (45%), moderate in 56 (28%), and severe in 54 (27%). Clinical diagnosis at admission was bronchopneumonia (38%), pyrexia of undetermined etiology (17%), bronchiolitis (13%), urinary tract infection (9%), acute gastroenteritis (8%), hepatitis (6.5%), and septicemia (5%). Patients with mild neutropenia recovered within 1 week. Among 110 patients with moderate/severe neutropenia, 80 (73%) recovered in <3 weeks. Predictors of prolonged neutropenia were age younger than 18 months (P < .01), absolute neutrophils count (ANC) < 500/mm3 (P < .05), hemoglobin < 10 gm/dL (P < .05), and positive CMV serology (P < .01). CMV and EBV serology were positive in 34.5% and 7.3% of patients, respectively. ANCA was positive in 42.8% of patients with prolonged severe neutropenia. In conclusion, neutropenia is a frequent finding in Egyptian infants and children, usually mild and transient, and mainly associated with infection. CMV and EBV are associated with prolonged neutropenia. Immune neutropenia is a common cause of moderate/severe neutropenia in the first two years of life.

Outcome of childhood acute Lymphoblastic leukemia in Egyptian children: A challenge for limited health resource countries

Abstract In childhood acute lymphoblastic leukemia (ALL) the reported 5-year event-free survival (EFS) rates are as high as 80%. Since 2004, multiple Egyptian centers shifted protocol of therapy of ALL to the CCG 1991 (the single delayed intensification arm) and CCG 1961 protocol for standard risk and high-risk ALL therapy, respectively, being cost effective. We aimed to evaluate the efficacy and safety of the CCG protocol in treatment of childhood ALL in Ain Shams and Menoufeya University hospitals. Methods Fifty-two ALL patients, aged 1–17 years, treated according to the modified CCG protocol in both centers and registered from November 2004 to December 2005 were included. They were classified into three risk groups, standard risk (SR), high-risk standard arm (HR-SA), and high-risk augmented arm (HR-AA). Results The mean age at diagnosis was 5.9 + 3.3 years, male/female ratio of 1.6:1, and central nervous system leukemia represented 6%. The 5-year overall survival (OS) and EFS were 84.6% and 67%, respectively. The 5-year OS and EFS were 92.6% and 70% in SR, 68.8% and 55% in HR-SA, 88.9% and 80% in HR-AA patients, respectively. Six patients had grade 3–4 adverse events. Conclusion The outcome of HR-SA protocol was inferior to the other two groups, necessitating shift to a more intensified arm with double delayed intensification. The use of minimal residual disease for better risk classification of childhood ALL is recommended in our centers

Thrombopoietin mutation in congenital amegakaryocytic thrombocytopenia treatable with romiplostim

Congenital amegakaryocytic thrombocytopenia (CAMT) is an inherited disorder characterized at birth by thrombocytopenia with reduced megakaryocytes, which evolves into generalized bone marrow aplasia during childhood. Although CAMT is genetically heterogeneous, mutations of MPL, the gene encoding for the receptor of thrombopoietin (THPO), are the only known disease‐causing alterations. We identified a family with three children affected with CAMT caused by a homozygous mutation (p.R119C) of the THPO gene. Functional studies showed that p.R119C affects not only ability of the cytokine to stimulate MPL but also its release, which is consistent with the relatively low serum THPO levels measured in patients. In all the three affected children, treatment with the THPO‐mimetic romiplostim induced trilineage hematological responses, remission of bleeding and infections, and transfusion independence, which were maintained after up to 6.5 years of observation. Recognizing patients with THPO mutations among those with juvenile bone marrow failure is essential to provide them with appropriate substitutive therapy and prevent the use of invasive and unnecessary treatments, such as hematopoietic stem cell transplantation or immunosuppression.

Determinants of platelet count in pediatric patients with congenital cyanotic heart disease: Role of immature platelet fraction

OBJECTIVES Congenital heart defects are common noninfectious causes of mortality in children. Bleeding and thrombosis are both limiting factors in the management of such patients. We assessed the frequency of thrombocytopenia in pediatric patients with congenital cyanotic heart disease (CCHD) and evaluated determinants of platelet count including immature platelet fraction (IPF) and their role in the pathogenesis of thrombocytopenia. METHODS Forty-six children and adolescents with CCHD during pre-catheter visits were studied; median age was 20.5 months. Complete blood count including IPF as a marker of platelet production and reticulated hemoglobin content (RET-He) as a marker of red cell production and iron status were done on Sysmex XE 2100 (Sysmex, Japan). C-reactive protein, prothrombin time (PT), Activated partial thromboplastin time (APTT) were also assessed. RESULTS Thrombocytopenia was found in 6 patients (13%). PT was prolonged (P = .016) and IPF was significantly higher in patients with thrombocytopenia compared with patients with normal platelet count (14.15 ± 5.2% vs 6.68 ± 3.39%; P = .003). Platelet count was negatively correlated with IPF while significant positive correlations were found between IPF and hemoglobin, red blood cells (RBCs) count, hematocrit (Hct), PT, reticulocytes count, and immature reticulocyte fraction. CONCLUSIONS We suggest that elevated IPF in CCHD patients with thrombocytopenia may denote peripheral platelets destruction as an underlying mechanism. Hemoglobin level, RBCs count, Hct, and RET-He were not significant determinants for platelet count in CCHD.

Alterations of platelet functions in children and adolescents with iron-deficiency anemia and response to therapy

Abstract Several changes in platelets have been reported in patients with iron-deficiency anemia (IDA), so a relationship between iron metabolism and thrombopoiesis should be considered. We aimed to study the alterations of platelet functions in patients with IDA by assessment of platelet aggregation with epinephrine, adenosine diphosphate (ADP) and ristocetin and by measuring platelet function analyzer-100 (PFA-100) closure time together with the effect of iron therapy on the same tests. A follow-up study was conducted in Ain Shams University Children’s hospital in the period from June 2011 to June 2012 including 20 patients with confirmed IDA and 20 healthy age- and sex-matched control. Bleeding manifestations were reported. Laboratory analysis included complete blood count, assessment of iron status by measuring serum iron, TIBC and ferritin, assessment of platelet functions by PFA-100 closure time and platelet aggregation with collagen, ADP and ristocetin. Patients with IDA were treated by oral iron therapy 6 mg/kg/day of ferrous sulfate and post-therapeutic re-assessment was done. Mean age of IDA patients was 5.7 ± 4.2 years. Bleeding manifestations were more common in patients group. Mean PFA-100 closure times (with epinephrine) were significantly longer in patients (179.1 ± 86.4 seconds) compared to control group (115 ± 28.5 seconds) (p < 0.05). Platelet aggregation by ADP (38.1 ± 22.2%), epinephrine (19.7 ± 14.2%) and ristocetin (58.8 ± 21.4%) were significantly reduced in patients compared to control (62.7 ± 6.2, 63.3 ± 6.9, 73.8 ± 8.3, respectively; p < 0.001). After treatment platelet aggregation tests induced by ADP (64.78 ± 18.25%), and epinephrine (55.47 ± 24%) were significantly increased in patients with IDA compared to before treatment (39.44 ± 21.85%, 20.33 ± 14.58%; p < 0.001). PFA-100 closure time as well showed significant decreased after treatment (118.4 ± 27.242) compared to before treatment (186.2 ± 90.35; p < 0.05). A negative correlation between platelet aggregation induced by ADP and mean values of serum ferritin before treatment (r = 0.042, p < 0.05) was found. A mutual effect is considered between iron deficiency and platelet functions. Subtle bleeding manifestations can occur in patients with IDA with delay in platelet aggregation and prolongation in PFA-100 closure times which can be reversed by iron therapy.

Clinically significant red blood cell antibodies in multitransfused Egyptian thalassemic patients

Background Red blood cell (RBC) alloimmunization is a major challenge to repeated transfusions in β-thalassemia-major patients. The aim of this study was to evaluate the magnitude of RBC alloimmunization and autoimmunization in regularly transfused Egyptian patients with β-thalassemia major and analyze factors that may be responsible for the development of antibodies. Patients and methods This study was conducted on 200 Egyptian β-thalassemia-major patients with age less than 16 years, who routinely visited the Pediatric Hematology Clinic of Ain Shams University Hospital for regular transfusions. All the patients underwent antibody screening. Patients with a positive antibody screen were further tested for antibody identification. The data were analyzed to find out the frequency of alloimmunization, and the patients′ records were revised to analyze the factors influencing it. In case of pan-positivity of the antibody-screening cells and the autocontrol, adsorption by autogenic RBCs was performed to uncover the presence of alloantibodies. Results RBC alloantibodies were found in 21 (10.5%) patients. The most frequent alloantibodies encountered were anti-Kell (52.4%) and anti-E (19%). Autoantibodies were encountered in only one patient. They were the warm autoantibody type, and adsorbtion using autogenic RBCs was succesful in eliminating them. The frequency of blood transfusion, the transfusion index, serum ferritin, and the age at first transfusion showed a statistically significant correlation with alloimmunization ( P < 0.05). In contrast, there was no statistically significant association between the patients′ sex, age, the ABO, Rh blood groups, and the spleen state and alloimmunization (P > 0.05). Conclusion We conclude that alloimmunization to RBC antigens is a relatively frequent finding among Egyptian transfusion-dependent thalassemic patients. The most frequent antibodies detected were against the Kell and Rh blood groups, mainly anti-Kell and anti-E. The majority of alloantibodies detected in this study were clinically significant.

Effect of Using Bedside Leukocyte Filter on Pulmonary Functions in Patients with Thalassemia Major

In settings of limited health resources, using leukocyte-filtered blood is limited to patients with leukocyte-mediated complications. The aim of this study was to determine the patterns of lung dysfunction among patients with β-thalassemia major (BTM) after the application of the leukostop filter during transfusion for a period of 6 months. The study included 30 patients with transfusion-dependent BTM divided into two groups according to the use of leukocyte filter. Group I included 15 patients with BTM allocated to use the leukocyte filter before each blood transfusion for 6 months and group II included 15 patients with BTM using nonleukocyte-filtered blood. Patients with history of airway disease and smokers were excluded. Chest X-ray and pulmonary function tests (PFT) using spirometry were done for each patient at baseline and after the use of the leukocyte filter for 6 months. No significant difference was found at baseline PFTs in both groups, the distribution of obstructive pulmonary disease significantly improved in group I in the postfilter evaluation, P < 0.05, however no change in pulmonary disease distribution in group II. A statistical significance improvement in forced vital capacity (FVC), forced expiratory volume in 1st second (FEV1) and FEV1/FVC in postfilter evaluation, while in group II a decline in FEV1, FVC, and no significant change in FEV1/FVC ratio. There was no correlation between serum ferritin and PFT results. Conclusion: Pulmonary function abnormalities, although subclinical is not an infrequent finding in patients with BTM; leukofiltred blood may improve PFT.

Effect of Honey Supplementation on Clostridium Difficile Infection in Childhood Cancer

Background: the treatment of cancer is associated with nausea and vomiting, oral mucositis, constipation, xerostomia and diarrhea and weight loss, additionally; chemotherapeutic agents promote inflammatory changes in the gut, intestinal necrosis, and anaerobic conditions, allowing proliferation of Clostridium Difficile. Honey, as a natural honeybee product, has antioxidant, antimicrobial, immunomodulatory and anticancer effects. Honey can fight microbial infection by its immuno-activating, anti-inflammatory and prebiotic activity. Objectives: the aim of this study was to evaluate the effect of honey supplementation on frequency of Clostridium Difficile infection (CDI) and gastrointestinal complications in pediatric patients undergoing chemotherapy. Design: a cross sectional study conducted on 40 patients with malignancy recruited from Childrens Hospital, Ain Shams University, Oncology Unit and Clinic, Cairo, Egypt in the period from December 2015 to December 2016. Patients were divided into two groups; group I (25 patients) received honey in the dose of 2gm/kg 3 times dailyfor 1month) while group II (15 patients) did not receive honey. All the studied patients were subjected to medical history and clinical examination, with special emphasis on gastrointestinal complication including oral mucositis, vomiting, diarrhea, constipation, and abdominal pain. Follow up was done for weight, height z score, gastrointestinal complications and any adverse events. Stool analysis, culture, C difficle toxin A, B by ELISA was done to all patients at baseline and repeated to patients receiving honey at week 4 of supplementation. Main outcome measure frequency of CDI, gastrointestinal complication, febrile neutropenia . Results: the frequency of C difficle was 8% (2), the first case was 9 years old patient with ALL (50%) and the other 11 years old patient with Burkitts lymphoma both were diagnosed by positive stool culture and positive stool ELISA for toxin A, B. gastrointestinal complications were significantly less and improved in the supplemented group and mean of hemoglobin significant increase in group 1. Conclusion: the frequency of CDI in children with cancer 8% diagnosed by stool culture and toxin A, B study in stool. Honey improved the oral mucositis and different GIT complications associated with chemotherapy.