Amany M. Ali

Immunophenotyping and immunoglobulin heavy chain gene rearrangement analysis in cerebrospinal fluid of pediatric patients with acute lymphoblastic leukemia

The study aimed to assess the diagnostic accuracy of Flow cytometry (FCM) immunophenotyping and IgH gene rearrangements (IGHRs) by real-time PCR in comparison with classic cytology for diagnosing CNS infiltration in pediatric ALL. We concluded that the diagnostic value of FCM and IGHR are two to three times more than that of cytology. Therefore, immunophenotyping by FCM is recommended for routine diagnosis of CSF infiltration. Furthermore, IGHR analysis by real-time PCR appears to be a useful addition in evaluation of CNS infiltration.

Incidence and characteristics of HBV reactivation in hematological malignant patients in south Egypt

AIM To investigate characteristics of hepatitis B virus (HBV) implicated in HBV reactivation in patients with hematological malignancies receiving immunosuppressive therapy. METHODS Serum samples were collected from 53 patients with hematological malignancies negative for hepatitis B surface antigen (HBsAg) before the start of and throughout the chemotherapy course. HBV reactivation was diagnosed when the HBsAg status changed from negative to positive after the initiation of chemotherapy and/or when HBV DNA was detected by real-time detection polymerase chain reaction (RTD-PCR). For detecting the serological markers of HBV infection, HBsAg as well as antibodies to the core antigen (anti-HBc) and to the surface antigen were measured in the sera by CEIA. Nucleic acids were extracted from sera, and HBV DNA sequences spanning the S gene were amplified by RTD-PCR. The extracted DNA was further subjected to PCR to amplify the complete genome as well as the specific genomic sequences bearing the enhancer II/core promoter/pre-core/core regions (nt 1628-2364). Amplicons were sequenced directly. RESULTS Thirty-five (66%) of the 53 HBsAg-negative patients were found to be negative serologically for anti-HBc, and the remaining 18 (34%) patients were positive for anti-HBc. Five of the 53 (9.4%) patients with hematologic malignancies experienced HBV reactivation. Genotype D1 was detected in all five patients. Four types of mutant strains were detected in the S gene product of HBV strains and were isolated from 3 patients with HBV reactivation: T/S120, L143, and I126. HBV DNA was detected in the pretreatment HBsAg-negative samples in one of the five patients with HBV reactivation. In this patient, sequences encompassing the HBV full genome obtained from sera before the start of chemotherapy and at the time of de novo HBV hepatitis were detected and it showed 100% homology. Furthermore, in the phylogenetic tree, the sequences were clustered together, thereby indicating that this patient developed reactivation from an occult HBV infection. CONCLUSION Past infection with HBV is a risk factor for HBV reactivation in Egypt. Mandatory anti-HBc screening prior to chemotherapy in patients with hematological malignancies is recommended.

Treatment results and prognostic factors of pediatric neuroblastoma: a retrospective study

Background We conducted a retrospective analysis to investigate treatment results and prognostic factors of pediatric neuroblastoma patients. Methods This retrospective study was carried out analyzing the medical records of patients with the pathological diagnosis of neuroblastoma seen at South Egypt Cancer Institute, Assiut University during the period from January 2001 and January 2010. After induction chemotherapy, response according to international neuoblastoma response criteria was assessed. Radiotherapy to patients with residual primary tumor was applied. Overall and event free survival (OAS and EFS) rates were estimated using Graphed prism program. The Log-rank test was used to examine differences in OAS and EFS rates. Cox-regression multivariate analysis was done to determine the independent prognostic factors affecting survival rates. Results Fifty three cases were analyzed. The median follow-up duration was 32 months and ranged from 2 to 84 months. The 3-year OAS and EFS rates were 39.4% and 29.3% respectively. Poor prognostic factors included age >1 year of age, N-MYC amplification, and high risk group. The majority of patients (68%) presented in high risk group, where treatment outcome was poor, as only 21% of patients survived for 3 year. Conclusion Multivariate analysis confirmed only the association between survival and risk group. However, in univariate analysis, local radiation therapy resulted in significant survival improvement. Therefore, radiotherapy should be given to patients with residual tumor evident after induction chemotherapy and surgery. Future attempts to improve OAS in high risk group patients with aggressive chemotherapy and bone marrow transplantation should be considered.

The Outcome of Critically Ill Pediatric Cancer Patients Admitted to the Pediatric Intensive Care Unit in a Tertiary University Oncology Center in a Developing Country: A 5-Year Experience.

Introduction: Cancer remains a major cause of death in children, but recent advances in supportive care and progress in the use of chemotherapy have considerably improved the prognosis. The need for intensive care management in pediatric oncology patients is increasing. However, studies demonstrating their outcome in the literature are still deficient, especially in developing countries. Here, we aim to report our experience in managing patients admitted to the pediatric intensive care unit (PICU) at South Egypt Cancer Institute, a tertiary university oncology center in a developing country. Patients and Methods: A review of all cancer patients admitted to the PICU at South Egypt Cancer Institute between January 2007 and December 2011 and an evaluation of prognostic factors that may correlate to their short-term outcome were performed. Results: A total of 550 pediatric oncology patients were admitted to the PICU on 757 occasions. Hematological malignancies represented 73.6% of the cases. The median duration of PICU stay was 5 days. Sepsis and respiratory failure were the most frequent indications for PICU admission. The overall survival at the time of discharge from the PICU was 60%. Several factors were found to significantly affect the outcome of patients admitted to the PICU, including the underlying disease, the reason for admission, the intervention used, and the number of failing organs at the time of admission to the PICU. Conclusions: The prognosis of patients admitted to the PICU in developing countries is still behind those in developed ones. Late referral, especially of patients presenting with respiratory failure, sepsis, and multiorgan failure usually, requires urgent intervention with inotropic support, oxygen therapy, and mechanical ventilation and is significantly associated with poor outcomes, especially in patients with hematological malignancies.

Characteristics of escape mutations from occult hepatitis B virus infected patients with hematological malignancies in South Egypt.

AIM To investigate the prevalence and virological characteristics of occult hepatitis B virus (HBV) infections in patients with hematological malignancies in South Egypt. METHODS Serum samples were collected from 165 patients with hematological malignancies to monitor titers of HBV DNA, hepatitis B surface antigen (HBsAg), and antibodies to HBV core (anti-HBc) and surface antigens. Serum samples negative for HBsAg and positive for anti-HBc were subjected to nucleic acid extraction and HBV DNA detection by real-time polymerase chain reaction. DNA sequences spanning the S region were analyzed in cases with occult HBV infection. In vitro comparative study of constructed 1.24-fold wild type and S protein mutant HBV genotype D clones was further performed. RESULTS HBV DNA was detected in 23 (42.6%) of 54 patients with hematological malignancies who were HBsAg negative, but anti-HBc positive, suggesting the presence of occult HBV infection. The complete HBV genome was retrieved from 6 occult HBV patients, and P120T and S143L were detected in 3 and 2 cases, respectively. Site directed mutagenesis was done to produce 1.24-fold genotype D clones with amino acid mutations T120 and L143. The in vitro analyses revealed that a lower level of extracellular HBsAg was detected by chemiluminescence enzyme immunoassay (CLEIA) with the clone containing T120 mutation, compared with the wild type or the clone with S143L mutation despite the similar levels of extracellular and intracellular HBsAg detected by Western blot. Southern blot experiments showed that the levels of intracellular HBV DNA were not different between these clones. CONCLUSION Occult HBV infection is common in patients with hematological malignancies and associated with P120T and S143L mutations. 120T mutation impairs the detection of HBsAg by CLEIA.

Role of Surgery in Stages II and III Pediatric Abdominal Non-Hodgkin Lymphoma: A 5-Years Experience

Abdominal Non-Hodgkin lymphomas (NHL) are the most common extra nodal presentation of pediatric NHL. Our aim is to assess the role of surgery as a risk factor and to evaluate the impact of risk-adjusted systemic chemotherapy on survival of patients with stages II and III disease. This study included 35 pediatric patients with abdominal NHL treated over five years at South Egypt Cancer Institute (SECI), Assiut University, between January 2005 and January 2010. The data of every patient included: Age, sex, and presentation, staging work up to determine extent of the disease and the type of resection performed, histopathological examination, details of chemotherapy, disease free survival and overall survival. The study included 25 boys and 10 girls with a median age of six years (range: 2.5:15). Thirty patients (86%) presented with abdominal pain, 23 patients (66%) presented with abdominal mass and distention, 13 patients (34%) presented with weight loss, and intestinal obstruction occurred in six patients (17%). The ileo-cecal region and abdominal lymph nodes were the commonest sites (48.5%, 21% respectively). Burkitts lymphoma was the most common histological type in 29 patients (83%). Ten (28.5%) stage II (group A) and 25 (71.5%) stage III (group B). Complete resections were performed in 10 (28.5%), debulking in 6 (17%) and imaging guided biopsy in 19 (54%). A11 patients received systemic chemotherapy. The median follow up duration was 63 months (range 51-78 months). The parameters that significantly affect the overall survival were stage at presentation complete resection for localized disease. In conclusion, the extent of disease at presentation is the most important prognostic factor in pediatric abdominal NHL. Surgery is restricted to defined situations such as; abdominal emergencies, diagnostic biopsy and total tumor extirpation in localized disease. Chemotherapy is the cornerstone in the management of pediatric abdominal NHL.

Risk-based combined-modality therapy of pediatric Hodgkin's lymphoma: A retrospective study

We conducted a retrospective analysis to investigate the clinical outcome of combined-modality therapy using multiagent chemotherapy and involved-field radiotherapy in treatment of children with Hodgkins lymphoma. Fifty eight cases with newly diagnosed Hodgkins lymphoma were analyzed. The median follow-up duration was 46 months (range 3-72 months). The 4-year overall and event-free survival rates were 91.5% and 69.7% respectively. High-risk disease (stage IIIB and IV), presence of B symptoms, lymphocyte depletion subtype, bulky disease and late response to chemotherapy were poor prognostic factors. Stage-adapted combined-modality therapy resulted in satisfactory outcome in treatment of pediatric Hodgkins lymphoma.

Risk-Adapted, Combined-Modality Therapy for Unfavorable Pediatric Hodgkin Lymphom

Background and Objectives: Risk-adapted therapy for children with HL is directed toward high survival, minimal toxicity and optimal quality of life, with long term follow up. We assess the impact of prognostic factors associated with local treatment failure of pediatric HL patients with unfavorable criteria treated with combined modality: Alternating ABVD (Doxorubicin, Bleomycin, Vinblastine and Decarbazine) and COEP (Cyclophosphamide, Oncovin, Etoposide and Prednisone) chemotherapy and response-based, involved-field radiation for newly diagnosed unfavorable pediatric HL patients, also will detect toxicities and long-term complications observed in the patients. Methods: This prospective study was carried out from January 2010 to January 2018, with a median follow up of 74 months (range 8 - 103 months). 54 patients were eligible for this study stratified into two groups: intermediate risk (IR) and high-risk group (HR). Patients were treated with (4 - 6 cycles) and (6 - 8 cycles) respectively of alternating ABVD/COEP chemotherapy followed by involved-field radiation therapy (IFRT): 15 Gy for patients achieved complete response, and 25.5 Gy for those achieved a partial response. Results: 27 patients were IR and 27 patients were HR. There were 16 treatment failures; 5 patients had progressive disease; and 11 patients had a relapse. 9 patients died from their disease progression. The 5-year overall survival (OS) and event-free survival (EFS) rates (±SE) were 81.8% ± 5.7% and 71.8% ± 6.2% respectively. Multivariate analysis revealed that the only independent factor for inferior OS was radiotherapy. Conclusion: Treatment results of unfavourable HL patients in our study are satisfactory for with IR group but not for HR group who needs intensification of therapy. Radiotherapy is considered as a cornerstone in the treatment of the patients with unfavourable criteria with better assessment of early responders needed by PET-CT to identify patients at risk for relapse.

Outcome and Clinical Significance of Immunophenotypic Markers Expressed in Different Treatment Protocols of Pediatric Patients With T-ALL in Developing Countries

Micro‐Abstract This study aimed to evaluate pediatric patients had T‐cell acute lymphoblastic leukemia (T‐ALL) at two different Arabic cancer centers regarding their clinic‐pathologic, immunophenotypic and cytogenetic features and outcome. Study included 103 patients with median age of 8.9 years. Male to female ratio was 2.6:1. Patients divided into (group I) treated with BFM‐90 treatment protocol between February 2003 and June 2007 and (group II) includes all patients treated thereafter by the total therapy study XIII protocol for high‐risk ALL. Outcome of patients with T‐ALL significantly improved in patients received treatment protocol of ALL with high‐risk criteria. This protocol eliminates the bad outcomes effect of several clinical and immunophenotypic markers. Background: T‐cell acute lymphoblastic leukemia (T‐ALL) accounts for about 15% of pediatric ALL. With wider use of intensive chemotherapy, the prognosis for childhood T‐ALL has improved. Further gains in treatment outcome will likely require methods to identify patients who continue to fail on contemporary protocols. This study aimed to evaluate pediatric patients with T‐ALL at 2 different Arabic cancer centers regarding their clinicopathologic, immunophenotypic, and cytogenetic features and outcome. Patients and Methods: This retrospective study included all children with T‐ALL treated between 2003 and 2013 at 2 oncology centers in the Middle East. Patients were divided into (group I) treated with Berlin‐Frankfurt‐Münster (BFM)‐90 treatment protocol between February 2003 and June 2007 and (group II) includes all patients treated thereafter by the Total Therapy Study XIII protocol for high‐risk ALL. Results: This study included 103 patients with a median age of 8.9 years. The male to female ratio was 2.6:1. The median initial white blood cell count was 123 × 109/L. Central nervous system leukemia was detected in 15%. The early T‐cell precursor (ETP)‐ALL phenotype was found in 16.5%. The 5‐year overall survival was 20.7% ± 67.5% and 72.9% ± 5.7% (P < .01); the 5‐year disease‐free survival was 47.1% ± 13.8% and 77.3% ± 6.0% (P = .023); and the 5‐year event‐free survival was 28.6% ± 12.1% and 71.1% ± 6.2% (P = .003) for group I and II, respectively. Conclusion: The outcome of patients with T‐ALL significantly improved in patients who received the treatment protocol of ALL with high‐risk criteria. This protocol eliminates the bad outcomes effect of several clinical and immunophenotypic markers. Patient with the ETP‐ALL phenotype had a nonsignificant inferior outcome compared with the non–ETP‐ALL group.

Sacrococcygeal tumors: clinical characteristics and outcome of pediatric patients treated at South Egypt Cancer Institute. A retrospective analysis.

BACKGROUND Sacrococcygeal tumors (SCT) are relatively uncommon tumors affecting neonates, infants and children. The aim of this article is to clarify any special characterizations in natural history, clinical presentation and outcome of such tumors treated at South Egypt Cancer Institute, the only research center located in South Egypt. METHODS A retrospective analysis of children with SCT treated at the Pediatric Oncology department South Egypt Cancer Institute, Assiut University between 2004 and 2010. RESULTS Nineteen children were included in the study. Age ranged between 10 days and 5 years. All but three had sacral mass at presentation. AFP levels ranged between normal age-related levels and 217,200 ng/ml. Initial resection was possible in 11, while eight patients with clinical suggestion of advanced malignant disease were inoperable. They received initial chemotherapy followed by delayed surgery. Yolk sac tumor (YST) was reported in 52.9% of patients. Recurrence was reported in 5 patients (3 mature teratomas and 2 YST). Five-year OS and RFS rates of patients who had malignant disease were 81.8% and 77.8% respectively. CONCLUSIONS Older age and delay in presentation that resulted in predominance of extensive disease and malignant transformation at presentation were the main challenges we faced in managing patients with SCT in our locality.