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  Citation statistics : Table of Contents
   2014| April-June  | Volume 39 | Issue 2  
    Online since August 30, 2014

 
 
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ORIGINAL ARTICLES
Flow cytometric evaluation of CD200 as a tool for differentiation between chronic lymphocytic leukemia and mantle cell lymphoma
Dahlia A El-Sewefy, Dina A Khattab, Mohamed T.H. Sallam, Walaa Ali Elsalakawy
April-June 2014, 39(2):42-46
DOI:10.4103/1110-1067.139754  
Background In the majority of cases, flow cytometry enables the differentiation of chronic lymphocytic leukemia (CLL) from mantle cell lymphoma (MCL). However, the diagnosis of CLL becomes challenging when CD23 is not expressed by the leukemic cells or in cases of MCL expressing CD23. CD200 is a membrane glycoprotein expressed on a subset of T and B lymphocytes. Its expression has been observed on human myeloma, plasma cells, and CLL cells. Aim of the work We investigated the pattern of expression of CD200 in CLL and MCL patients, aiming to clarify its possible role in differentiating these often overlapping disorders. Patients and methods This study was carried out on 30 patients with newly diagnosed CLL and 10 patients with MCL. Flow cytometric immunophenotyping was performed using the CD200 monoclonal antibody in addition to the standard panel of chronic lymphoproliferative diagnosis. Results CD200 was expressed in 100% of CLL cases with moderate intensity compared with only 10% of MCL cases with low intensity. Conclusion CD200 is a powerful addition to the routine flow cytometric panel in the differentiation between CLL and MCL with high sensitivity and specificity.
  2 3,305 462
Study of T-regulatory cells in patients with acute, idiopathic thrombocytopenic purpura
Ahmad Baraka, Maher Borai, Mohamed A Hesham, Mohamed A.A. Almalky
April-June 2014, 39(2):37-41
DOI:10.4103/1110-1067.139751  
Introduction Idiopathic thrombocytopenic purpura (ITP) is an autoimmune bleeding disorder that occurs because of enhanced peripheral platelet destruction. Antibodies and T cells are involved in the pathogenesis of the disease and, like other autoimmune diseases, patients with ITP have a peripheral deficiency in regulatory T cells (Treg) numbers and function that may be responsible for loss of tolerance. Our aim was to measure Tregs (CD4 + CD25 +high FoxP + 3) and levels of interleukins (IL-10 and IL-12) in peripheral blood mononuclear cell (PBMC) cultures from patients with ITP and analyze their relationship with the clinical features and outcome of treatment of ITP. Participants and methods Forty-five participants were included in this study, divided into two groups. Group I included 15 healthy children as a control group. Group II included 30 pediatric patients with ITP. According to treatment, group II was divided into three subgroups: group IIa (no treatment) included two (6.7%) patients, group IIb (steroid treatment) included 10 (33.3%) patients, and group IIc (steroid+intravenous immunoglobulin treatment) included 18 (60%) patients. ITP is diagnosed by platelet count less than 100 Χ 103/μl. Tregs were analyzed by flow cytometry. IL-10 and IL-12 in the supernatants of basal and lipopolysaccharide-stimulated PBMC cultures were estimated using an enzyme-linked immunosorbent assay. Results A significantly lower percentage of Tregs was found in patients than in controls (1.46 ± 0.97 vs. 7.09 ± 1.5%) and the lowest percentage of Tregs was recorded in group IIc. A positive correlation was observed between Tregs% and platelet count in the patient group. PBMCs from patients had significantly higher basal levels of IL-10 and IL-12, with a marked reduction in responsiveness to lipopolysaccharide in vitro compared with the controls. Conclusion Children with ITP had reduced Tregs% and IL-10/IL-12 imbalance. Thus, Tregs may play a role in modifying immune responses in these patients, resulting in new strategies of treatment and monitoring of disease activity.
  1 2,045 269
Serum 25-hydroxyvitamin D levels in relation to disease status and prognosis in acute myeloid leukemia
Tahani A Elkerdany, Doaa G Eissa, Mohamed M Moussa
April-June 2014, 39(2):47-51
DOI:10.4103/1110-1067.139757  
Background Low vitamin D levels are linked to higher incidence of cancer. Although vitamin D insufficiency is related to inferior prognosis in some cancers, no data exist for acute leukemia. Aim of the work To evaluate the relationship between serum 25-hydroxyvitamin D (25[OH]D) levels and disease status in acute myeloid leukemia (AML) and correlate these levels with prognostic markers of the disease. Materials and Methods Sixty AML patients, 32 newly diagnosed, 15 in relapse, and 13 in complete remission, as well as 30 healthy control individuals were studied using enzyme-linked immunosorbent assay for the measurement of serum 25[OH]D. Results There was a significantly lower white blood cell count, hemoglobin level, platelet count, and serum vitamin D level among AML patients compared with the control group. Vitamin D-insufficient patients showed lower hemoglobin levels and platelet counts, together with a higher lactate dehydrogenase level, higher percentage of peripheral blood blasts infiltration, and higher percentage of bone marrow blasts' infiltration compared with vitamin D-sufficient patients. Vitamin D levels showed a significantly higher median value among patients who had favorable cytogenetics and a higher median value among those who showed complete remission than those newly diagnosed or in relapse. Survival curve analysis showed shorter overall survival of vitamin D-insufficient patients compared with vitamin D-sufficient patients. Conclusion and Recommendations Vitamin D level is related to the active stage of the disease and therefore aggressiveness of the disease. It can be used as a prognostic tool for survival in AML patients. It is recommended that further evaluation be carried out on a large number of patients with a longer follow-up period and that it is incorporated into the routine evaluation of all AML patients. Additional studies are needed to determine the exact role of vitamin D in acute leukemia and whether it could be used as a therapeutic or a preventive approach.
  - 1,755 209
High levels of soluble thrombomodulin may be a marker of arterial disease and peripheral ischemia in Egyptian patients with diabetes mellitus
Walaa A Elsalakawy, Botheina A.T. Farweez, Mohamed T.H. Sallam, Mohamed A Hamza
April-June 2014, 39(2):52-57
DOI:10.4103/1110-1067.139759  
Background Thrombomodulin is an endothelial cell membrane protein acting as a cofactor for the activation of plasma protein C. It was found that soluble forms of thrombomodulin (sTM) exist in plasma and is considered a marker of endothelial dysfunction. Materials and methods The study included 160 individuals: 40 healthy controls and 120 diabetic patients (40 patients with type 1 diabetes and 80 patients with type 2 diabetes). sTM concentrations were measured by an enzyme-linked immunosorbent assay-based assay employing monoclonal antithrombomodulin antibodies. Results The diabetic patients had highly significantly increased plasma sTM concentrations compared with the control group (P < 0.001). Type 2 diabetic patients with peripheral limb ischemia showed a highly significant increase in plasma sTM levels when compared with type 2 or type 1 diabetic patients without ischemia (P < 0.001). sTM concentrations were positively correlated with age, duration of diabetes, and urinary albumin concentration, with highly significant P value (P < 0.001). A cutoff value of greater than 9500 ng/ml of sTM was found to be of 96.9% diagnostic accuracy in differentiating type 2 diabetic patients with and without peripheral limb ischemia. Conclusion We recommend including sTM in the follow-up panel of diabetic patients because of its potential ability to predict the risk for ischemic manifestations.
  - 1,149 122
Impact of urinary schistosomiasis on haematological parameters and frequency of vaso-occlusive crisis among patients with sickle cell disease in northern Nigeria
Sagir G Ahmed, Modu B Kagu, Umma A Ibrahim
April-June 2014, 39(2):58-63
DOI:10.4103/1110-1067.139762  
Background Sickle cell disease (SCD) patients have impaired immunity with vulnerability to infections including parasitic infestations. Majority of SCD patients live in nations where urinary schistosomiasis is also endemic. This study was aimed at determining the impact of urinary schistosomiasis on haematological parameters and frequency of vaso-occlusive crisis (VOC) among patients with SCD. Materials and Methods We compared the haematological parameters and frequency of VOC among SCD patients with and without schistosomiasis in northern Nigeria. Result SCD patients with schistosomiasis had lower haematocrit, higher reticulocyte count, higher prevalence of iron deficiency, more intense leucocytosis and thrombocytosis, higher prevalence of bacterial urinary tract infections and higher ESR. The frequency of VOC was significantly higher in SCD patients with schistosomiasis. Conclusion The results of this study suggest that urinary schistosomiasis adversely affected the severity and prognosis of SCD. SCD patients, the majority of whom live in schistosomiasis endemic countries, should have regular urine tests for early detection and treatment of schistosomiasis in order to avert its adverse interaction with SCD.
  - 2,248 239
Study of MEFV gene R202Q polymorphism in Egyptian patients with familial Mediterranean fever
Lobna O ElMessery, Hala F Elhagrasy
April-June 2014, 39(2):64-67
DOI:10.4103/1110-1067.139764  
Familial Mediterranean fever (FMF) is a hereditary disorder characterized by recurrent attacks of fever and serositis. Several mutations in the MEFV gene have been associated with the disease. The aim of the present study is to investigate the frequency of the R202Q(605G>A) polymorphism in exon 2 of the MEFV gene and its clinical significance in a cohort of Egyptian patients with FMF. The study included 55 FMF patients and 40 healthy controls. The polymorphism was tested using polymerase chain reaction (PCR) based restriction fragment length polymorphism (RFLP). The genotype and allele frequency was similar in patients and controls. No significant association was found between demographic and clinical characteristics of patients and R202Q genotype. Further studies covering wider geographical areas in Egypt are required to reveal the diagnostic relevance of this molecular marker in Egypt.
  - 1,561 138
Regulatory T cells in chronic lymphocytic leukemia
Salah Aref, Tawfik Elkhodary, Emad Azmy, Hazem Hakim, Nadia El Menshawy, Lamiaa Ebrahim
April-June 2014, 39(2):68-71
DOI:10.4103/1110-1067.139766  
Background: Controversial results concerning the prognostic value of the Tregs cells percentage in peripheral blood of chronic lymphocytic leukemia (CLL) patients have been reported in the previous studies. Materials and Methods: This study aimed to estimate the prognostic relevance of Tregs cells in untreated CLL patients at diagnosis. Results: CLL patients showed significantly higher Tregs percentage and Tregs cell counts as compared that identified in healthy normal controls (p < 0.01 for both). Furthermore, CLL patients with high LDH, β2 microglobulin levels, those positive for CD38% express significantly higher Tregs percentage as compared to those patients with normal LDH, β2 microglobulin levels and negative CD 38%. Also, the percentage of Tregs cells was significantly higher among CLL patients having autoimmune cytopenias. Conclusion: Tregs cells percentage is higher in CLL patients as compared to normal healthy controls and related to advanced stages as well as poor prognostic markers. Tregs manipulation may represent a future strategy for management of CLL patients.
  - 1,380 133
Study of the role of HSP90 in acute myeloid leukemia
Sahar M Hazza, Aml A Azzat, Ola A El Shora, Mohammed M El-Bedwey
April-June 2014, 39(2):72-79
DOI:10.4103/1110-1067.139769  
Review Heat shock proteins (HSPs) are a group of proteins whose expression are increased when the cells are exposed to elevated temperature or other stress. This increase in expression is transcriptionally regulated. A member of HSP family is HSP90, it is expressed in cytoplasm of most human cells.HSP90 exists in two forms HSP90 alpha, inducible and HSP90B, constitutive. HSP90 has a more restricted repertoire of client proteins mainly including protein kinase. It seems to be important for cellular proliferation, survival and adaptation to unfavorable microenvironments, and HSP90 inactivation results in inappropriate functioning and rapid degradation of its client proteins. Its normal chaperoning activity is strongly dependent on its co-chaperones, i.e. accessory proteins that interact with HSP90 and induce conformational changes. Aim The work was conducted to to evaluate the role of HSP90 in acute myeloid leukemia and find out its impact on prognosis. Subject and Method The subjects of this study were 90 patients with acute myeloid leukemia. The age of patients ranged from 4-67 years with a mean value of 43.367 ±15.146 years. They were 66 (73.33%) males, and 24 (26.67%) females. After diagnosis patients received chemotherapy and they were followed up for periods of 18 months with special attention to clinical and laboratory markers of remission and relapse and estimation of ate of first complete remission, date of relapse, death or last seen alive. Results In the current study, 90 newly diagnosed acute myeloid leukemic patients were investigated for HSP90 expression. High expression of HSP90 (>20%) was found in 48/90 (53.33%) AML patients and low expression of HSP90 (<20%) was found in 42/90 of patients (46.67%). In the current work, the age of selected patients ranged from 4-67 years with mean age of 43 years. In present study, it was found that 66/90 AML patients (73.33%) were males and 24/30 AML patients (26.67%) were females. Among the newly diagnosed AML patients; 43.33% had hepatomegaly, 50% had lymphadenopathy, 50% had pallor, 40% had purpura, and 60% had splenomegaly. There was no correlation between HSP90 expression and clinical features. As regard hematological finding, there was no significant association between HSP90 and Hb level. The present study showed mild to severe thrombocytopenia in almost all AML patients, There was no association between platelet count and HSP90 expression. There was a significant association of high total leucocytic count with high HSP90 expression. Circulating blasts in peripheral blood and blasts in BM were significantly associated with high expression of HSP90 in AML patients compared to low expressed HSP90 cases. There was statistically non significant relation between HSP90 expression and FAB subtype. There was a statistically significant association between high HSP90 expression and bad prognosis including relapse or death while the high remission rates were associated significantly with low expression of HSP90 cases.
  - 1,290 158
Prognostic utility of CD184 in acute myeloid leukemia
Mohamed A Attia, Sahar M Hazzaa, Ola A El-Shora, Tamer A El-Bedewy, Loi M El-Ahwal
April-June 2014, 39(2):80-85
DOI:10.4103/1110-1067.139772  
Background Chemokine induce directional migration of cells toward a gradient of chemotactic cytokines (chemotaxsis). CD184 is a chemokine receptor which regulates the localization of leukemic cells, and most leukemic cells respond to it, with increased adhesion, survival and proliferation. Aim The work was conducted to evaluate the role of CD184 in newly diagnosed patients with acute myeloid leukemia. Subject and method Eighty patients presented with de novo AML. Based upon the distribution of CD184 expression in our patients' population, we defined groups with low CD184 expression as group I (MFIR 4 to 8), intermediate CD184 expression as group II (MFIR 9 to 20), and high CD184 expression as group III (MFIR more than 21). Patients with acute myeloid leukemia were studied at first diagnosis and were previously untreated. After diagnosis, patients received chemotherapy and they were followed up for periods ranged of 24 months with special attention to clinical and laboratory markers of remission and relapse, taking care to estimate the date of first complete remission, date of relapse, death or last seen alive. Results patients were divided according to CD184 expression (MFIR) into 3 groups; group (I) with low expression (MFIRs ≤9) had a mean of 5.04 ± 1.48, group (II) with intermediate expression (MFIRs between 9 to 20) had a mean of 10.03 ± 0.45, and group (III) with high expression (MFIRs ≥20) had a mean of 30.25 ± 6.17. In the present study, we found no relation between CD184 expression levels in AML patients and the age of patients. Also, no significant relation was found between CD184 expression levels and the presence or absence of hepatosplenomegally and lymphadenopathy (P > 0.05 ). As regards to haematological data and MFIR of CD184 expression in AML patients, the mean leukocytic count was significantly lower in group (I) than in group (II) and group (III), and leucocytic count was significantly related to CD184 expression in the three groups. Also, the percentage of blasts in peripheral blood and bone marrow was significantly related to CD184 expression with P value < 0.05 while no significant relation was found with haemoglobin or platelets (P > 0.05). As regards to prognosis and MFIR of CD184 expression, group (I) showed 70% remission, 20% relapse and 10% death, while group (II) showed 33.33% remission and 66.67% relapse and group (III) showed 100% deaths meaning that high and intermediate CD184 expression were associated with low rates of remission and high frequency of relapse and death. We can conclude that CD184 expression can represent a useful prognostic tool for AML patients.
  - 1,201 114
TPMT gene polymorphism detection by conventional PCR in pediatric acute lymphoblastic leukemia and its toxic effect
Dalal M.N.E. El-Kaffash, Hoda M.A.F. Hassab, Abla A AbouZeid, Rania S Swelem, Mona M Tahoun
April-June 2014, 39(2):86-90
DOI:10.4103/1110-1067.139775  
Introduction The purine analog mercaptopurine is a key medication for the successful treatment of childhood acute lymphoblastic leukemia, particularly for the consolidation and continuation therapies. Thiopurine S-methyltransferase (TPMT) catalyzes the inactivation of mercaptopurine. TPMT single-nucleotide polymorphisms can prospectively identify patients at higher risk for mercaptopurine toxicity. Patients and methods The TPMT genotype was determined by in-house conventional PCR followed by digestion of the product with restriction enzymes, MwoI FastDigest and AccI FastDigest. The study was carried out on a total of 80 participants: 40 pediatric patients with standard risk B-cell acute lymphoblastic leukemia and 40 age-matched and sex-matched healthy controls. Mercaptopurine was given to the patients in consolidation phase with oral dose of 75 mg/m 2 daily for 4 weeks. Toxicity of the drug was assessed at the end of this phase by complete blood profile and liver function tests. Results In the patients group, 97.5% were of the wild-type homozygous TPMT*1/*1 genotype and 2.5% were of the heterozygous TPMT*1/*3A genotype. In the control group, we identified 90% with the TPMT*1/*1 genotype, 7.5% with the TPMT*1/*3A genotype, and 2.5% with the TPMT*1/*3C genotype. Among the wild-type *1/*1 genotype patients in the patient group, 32.5% of patients suffered from either hepatoxicity and/or myelosuppression. Conclusion The homozygous wild-type TPMT*1/*1 genotype was the most frequent genotype in both cases and controls. TPMT*1/*3A was the most prevalent mutant genotype in this study. Although some patients had wild-type allele genotyping, they developed signs of toxicity.
  - 1,947 187