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   2013| January  | Volume 38 | Issue 1  
    Online since June 20, 2014

 
 
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ORIGINAL ARTICLES
Clinical relevance of thrombospondin receptor (CD36) expression in Egyptian de novo adult acute myeloid leukemia
Sherin M. Abd El-Aziz, Dalia A. Salem, Manal A. Salah-Eldin
January 2013, 38(1):1-6
DOI:10.7123/01.EJH.0000423008.85761.a3   
Background

Acute myeloid leukemias (AMLs) are a heterogeneous group of disorders that often present with different morphological, immunophenotypic, and cytogenetic patterns. Identification of these characteristics may be useful for a better prognostic evaluation and for a more appropriate therapeutic approach. CD36 is a transmembrane, highly glycosylated, glycoprotein commonly expressed on blasts in acute monocytic leukemia, megakaryoblastic leukemia, and erythroleukemia.

Patients and methods

We evaluated CD36 surface expression in 97 newly diagnosed AML patients, and the results were correlated with the morphology, immunophenotype, cytogenetic pattern, and clinical outcome.

Results

CD36 antigen was recorded in 48 of 97 patients (49.5%) and particularly in those with M5 and M6 FAB subtypes. Moreover, CD36 expression was significantly associated with the expression of CD11b (P=0.001) and CD14 (P=0.0001), unfavorable cytogenetic abnormalities (P=0.001), shorter overall survival (P>0.0001), and leukemia-free survival (P=0.03).

Conclusion

On the basis of the results of the study, it can be concluded that CD36 expression in AML patients may identify a subgroup with a poor prognosis, and may thus be a valuable adjunct to be added to the current prognostic factors.

  2,716 184 -
Evaluation of bone marrow examination in patients with hepatitis C virus infection
Inas A Asfour, Maryse S Ayoub, Nanees A.A. Magid, Ahmed A.I. Elsaharty
January 2013, 38(1):13-16
DOI:10.7123/01.EJH.0000423010.93384.6d   
Background

Over 170 million individuals are infected with hepatitis C virus (HCV) worldwide. The report published by the Egyptian Demographic Health Survey stated that in Egypt, there is an overall anti-HCV antibody prevalence of 14.7% and the number of Egyptians estimated to be chronically infected was 9.8%. Hematological manifestations are among the most common extrahepatic manifestations of HCV infection. Patients with HCV infection can develop peripheral blood cell count abnormalities that are commonly attributed to hypersplenism, antiviral therapy, decreased thrombopoietin levels, and/or autoimmune mechanisms.

Aim of the work

Evaluation of bone marrow findings in patients infected with HCV presenting with peripheral blood cytopenias.

Material and Methods

This study was carried out on 35 patients with chronic HCV infection presenting with cytopenias. Patients were subjected to history taking, physical examination, and routine laboratory investigations together with bone marrow aspiration and trephine biopsy. Flow cytometry and cytogenetics analysis were carried out in selected patients.

Results

Among the patients studied, B-cell non-Hodgkin lymphoma was the most commonly encountered disorder (31%), followed by idiopathic thrombocytopenic purpura (11%), acute lymphoblastic leukemia, acute myeloid leukemia, myelodysplastic syndrome, and hypersplenism (9% each), and autoimmune hemolytic leukemia (6%), and the least encountered disorders were anemia of chronic disease, aplastic anemia, CLL, myelofibrosis, Evan’s syndrome, and sideroblastic anemia (3% each).

Conclusion

B-cell non-Hodgkin lymphoma was the most frequently detected problem among the patients recruited (31%). There were significant data regarding relation of bone marrow dysplasia and fibrosis with age of patients.

  2,683 208 -
Cellular vascular endothelial growth factor and serum angiogenin in acute myeloid leukemia ( clinical and prognostic significance)
Mona A Ismail, Deena Samir Eissa, Nihal M Heiba
January 2013, 38(1):41-46
DOI:10.7123/01.EJH.0000423015.67835.8c   
Background

Increased angiogenesis and angiogenic factors play an important role in hematologic malignancies. Acute myeloid leukemia (AML) is associated with a considerable increase in bone marrow vascularity. Vascular endothelial growth factor (VEGF) and angiogenin (ANG) are two prominent angiogenic mediators. Therefore, we aimed to examine the cellular expression of VEGF and serum ANG level in adult AML patients and to assess their prognostic impact on disease outcome.

Materials and methods

The study was carried out on 60 newly diagnosed adult AML patients compared with 25 age-matched and sex-matched controls. Patients were diagnosed and classified according to the French–American–British/WHO criteria by immunophenotyping and cytogenetic analysis. The cellular expression of VEGF in blast cells was assessed by flow cytometry. Serum levels of ANG were measured using a sandwich enzyme-linked immunosorbent assay.

Results

AML blasts significantly overexpressed VEGF (median percentage expression, 30%; median mean fluorescence intensity, 3.8) compared with normal individuals (<5%; 0.57; P<0.01). Similarly, the serum levels of ANG were significantly higher in AML patients (mean±SD, 212.5±77.1 ng/ml) compared with the controls (mean±SD, 66±2.5 ng/ml; P<0.01). High levels of VEGF, above the median (>30%), together with serum ANG levels of AML patients showed no relationship with the studied clinical and laboratory parameters, French–American–British subtypes, or cytogenetic risk groups (P>0.05); however, they were significantly associated with a poor response to treatment and a higher mortality rate (P<0.01).

Conclusion

Cellular VEGF expression and serum ANG levels are significantly increased in AML patients and have been identified as independent poor prognostic indicators linked to accelerated angiogenesis and consequently a more aggressive disease outcome.

  1,979 405 -
Platelet aggregation in various stages of diabetic retinopathy ( evaluation using the PFA-100)
Deena M.M. Habashy, Mohamed R. Mohamed
January 2013, 38(1):17-22
DOI:10.7123/01.EJH.0000423011.70514.64   
Background

Platelet hyperactivity has been reported in diabetic patients. Some evidence suggests that platelet hyperaggregation may participate in the pathogenesis of diabetic complications such as retinopathy.

Objectives

We aimed to compare platelet aggregation (PA) in patients with type II diabetes with no apparent retinopathy (NAR), nonproliferative diabetic retinopathy (NPDR), proliferative diabetic retinopathy (PDR) patients, and healthy individuals to evaluate the possible role of PA in the pathogenesis and staging of DR.

Participants and methods

Blood samples from 30 patients (10 diabetics with NAR, 10 with NPDR, and 10 with PDR) and 10 healthy individuals were assayed for PA using a platelet function analyzer-100.

Results

On comparing all the studied groups in terms of the demographic, clinical, and laboratory parameters, the mean age and the duration of diabetes were significantly higher in the PDR group (P<0.001). The PDR group also showed higher levels of platelet count, glycated hemoglobin, and shorter closure time (CT) (P<0.001). Both NPDR and PDR showed higher levels of fibrinogen (P<0.001). CT was correlated inversely with the fibrinogen level in the NAR group (P=0.000) and with the duration of diabetes in the NPDR group (P=0.04). No correlation was found between CT and any of the parameters studied in the PDR group (P>0.05).

Conclusion

PA is increased in the PDR stage in comparison with NAR, NPDR stages, and healthy individuals. This may provide a clue of its role in the pathogenesis and staging of DR. Also, this study highlights the crucial contribution of glycemic control and duration of diabetes in the progression of DR.

  2,055 160 -
Frequency of human hemochromatosis (HFE) gene mutations in Egyptians with β-thalassemia
Doha A Mokhtar, Mona M Hamdy, Ahmed M Badr
January 2013, 38(1):36-40
DOI:10.7123/01.EJH.0000423014.60212.a7   
Background

Hereditary hemochromatosis is a disorder of iron metabolism characterized by increased iron intake and progressive storage and is related to mutations in the HFE gene. Two point mutations have been described and are referred to as H63D and C282Y. On the other hand, iron overload is a well-documented complication in thalassemia syndromes. Interactions between thalassemia and hemochromatosis may further increase iron overload. This work aimed at studying the frequency of the H63D and C282Y mutations of the HFE gene in an Egyptian population with β-thalassemia (thalassemia major, intermedia, and minor) by comparing it with normal individuals without hemoglobinopathies.

Participants and methods

This study included 86 patients with β-thalassemia; 40 of these patients had β-thalassemia major and intermedia and the other 46 patients had β-thalassemia minor (carriers). In addition, 70 individuals were included in the study and served as controls. All the populations studied were screened for H63D and C282Y mutations of the HFE gene using the PCR-restriction fragment length polymorphism (PCR-RFLP) method.

Results

The allelic frequencies found for H63D and C282Y mutations in this study were 18.6 and 0%, respectively, among the total alleles of individuals with β-thalassemia and 12.8 and 1.4% among controls without hemoglobinopathies. On comparing thalassemia cases, thalassemia carriers, and the control group, the allele frequency of the H63D mutation was significantly higher among β-thalassemia carriers (24%) compared with patients with β-thalassemia (12.5%) and controls (12.8%) (P=0.049). Our study also found an almost significantly higher frequency of the heterozygote genotype for the H63D mutation in patients with β-thalassemia (30.2%) than the controls (17.1%) (Fisher’s test, P=0.06). Compound heterozygosis for H63D and C282Y was found in one of the individuals in our control group.

Conclusion

The frequent occurrence of β-thalassemia disease with the H63D gene mutation raises the possibility of genetic interactions and emphasizes the value of screening for HFE mutations in thalassemias to detect early cases of iron overload and to modify its treatment modalities.

  1,978 192 -
Plasminogen activator inhibitor-1 4G/5G gene polymorphism in hemodialysis patients with cardiovascular disease
Baheia H. Mostafa, Doha A. Mokhtar, Ahmed M. Badr, Nasser M. Gamal el Din
January 2013, 38(1):29-35
DOI:10.7123/01.EJH.0000423013.60212.69   
Background

Plasminogen activator inhibitor-1 (PAI-1) exerts antifibrinolytic and profibrotic activities and it plays an important role in renal fibrosis. Moreover, PAI-1 is also considered as a risk factor for cardiovascular disease. A 4G/5G polymorphism of the PAI-1 gene has been described associating the 4G haplotype with higher PAI-1 plasma activity. The aim of this study was to examine the frequency and distribution of the 4G/5G PAI-1 genotypes in patients with end-stage renal disease (ESRD) who developed cardiovascular complications in the form of hypertension, echocardiographic changes, and vascular thrombosis and the possible link(s) between them.

Materials and methods

We studied 40 patients with ESRD who had cardiovascular complications: 20 patients on hemodialysis (50%), 10 on conservative treatment (25%), and 10 subjected to renal transplantation (25%), in addition to 30 healthy individuals who served as controls. Genotyping of the PAI-1 gene was performed using allele-specific PCR method.

Results

The homozygous 4G/4G genotype was more frequent than the other genotypes (heterozygous 4G/5G and wild 5G/5G) among patients when compared with controls with a statistically significant difference (P=0.01). A significant difference was also found on comparing the presence of the mutant 4G allele (in 4G/4G and 4G/5G genotypes) or its absence (in the 5G/5G genotype) between patients and controls (P=0.04). On studying the genotyping of the four different groups, we found that the 4G/4G genotype was more prevalent among hemodialysis group, the 4G/5G was more prevalent among transplanted group, whereas the 5G/5G genotype was more frequent in the control group, and these differences were highly significant statistically (P=0.005). For the genotype frequencies and their potential associations with cardiovascular complications and/or different laboratory findings, we only found a nearly significant association between the presence of the mutant 4G allele and lower high-density lipoprotein cholesterol levels (P=0.08). Among patients who were subjected to renal transplantation, all patients who developed cardiovascular complications (50%), increased creatinine (10%), or repeated graft rejections (40%) had the heterozygous genotype (4G/5G) with the mutant 4G allele.

Conclusion

We found that the 4G/4G genotype in addition to the mutant 4G allele was more frequent among patients with ESRD compared with controls. The presence of the 4G mutation showed a nearly significant association with lower high-density lipoprotein cholesterol levels, suggesting that it could play a role in the pathogenesis of accelerated atherosclerotic heart disease in uremic patients.

  1,881 132 -
Role of T-helper 17 cells and interleukin-17 expression in patients with acute myeloid leukemia
Mona A Ismail, Mohamed T.H. Sallam, Walaa A El Salakawy
January 2013, 38(1):47-50
DOI:10.7123/01.EJH.0000423016.44965.49   
Background

Several observations suggest that immunological events are important for antileukemic immune reactivity in acute myeloid leukemia (AML). T-helper type 17 (Th17) cells play an active role in inflammatory and autoimmune diseases; however, the nature of Th17 cells in hematological malignancies remains unknown.

Aim

The aim of this work was to assess the role of Th17 cells and its related cytokine interleukin-17 (IL-17) in the pathogenesis and prognosis of AML.

Participants and methods

The study was carried out on 40 adult AML patients and 20 age-matched and sex-matched controls. Patients were diagnosed and classified according to the WHO criteria for AML. Plasma levels of IL-17 in patients and controls were measured using the sandwich ELISA technique. Th17 cells were assessed using flow cytometry.

Results

The percentage of Th17 cells and the concentration of IL-17 were significantly increased in untreated patients with AML compared with the healthy controls (10.2±2.2 vs. 4.38±0.16% and 20.15±2.9 vs. 8.2±1.3 pg/ml, respectively). In addition, the increased Th17 cells were reduced significantly when the patient achieved complete remission after chemotherapy, but the decrease was not significant in patients with incomplete remission.

Conclusion

These results suggest that Th17 cells play a role in the pathogenesis and response of therapy in AML patients through the secretion of IL-17 cytokine.

  1,809 146 -
Growth differentiation factor 15 expression in anemia of chronic disease and iron deficiency anemia
Hala M.H. Abaza, Deena M.M. Habashy, Rana E.A. El-Nashar
January 2013, 38(1):23-28
DOI:10.7123/01.EJH.0000423012.78137.2e   
Background

Growth differentiation factor 15 (GDF15) expression, at the high levels achieved in the setting of ineffective erythropoiesis, contributes toward pathological iron overloading through a mechanism of incomplete hepcidin suppression. In addition to the regulation of hepcidin, iron depletion or chelation in the host may regulate GDF15 expression.

Objectives

This study aimed to detect GDF15 expression in Egyptian patients with iron deficiency anemia (IDA) and anemia of chronic disease (ACD) to examine its possible role in their differentiation.

Patients and methods

GDF15 was detected using an enzyme-linked immunosorbent assay in 40 patients (20 with IDA and 20 with ACD) and 10 age-matched and sex-matched healthy controls.

Results

The IDA group showed a decrease in the mean values of hemoglobin (Hb), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), serum iron and ferritin, and higher total iron-binding capacity (TIBC) compared with the controls (P⩽0.001). The ACD group had a statistically significantly higher mean value of GDF15 than the control and IDA groups (P⩽0.001). The mean values of Hb, MCV, serum iron, and TIBC were statistically significantly lower, with higher serum ferritin in the ACD group compared with the controls (P⩽0.001, P=0.01, P⩽0.001, P⩽0.001, and P=0.004, respectively). Higher mean values of MCV, MCH, and ferritin were found in the ACD compared with the IDA groups (P⩽0.001), whereas TIBC was higher in the IDA group compared with the ACD groups (P⩽0.001). No statistically significant correlation was found between the serum GDF15 level and the laboratory data studied among the ACD and IDA groups (P>0.05). No difference was found between serum C-reactive protein-positive and serum C-reactive protein-negative ACD groups in the laboratory parameters studied (P>0.05).

Conclusion

GDF15 plays an important role in the pathogenesis of ACD and IDA, and it can be used as a potential marker in their differentiation. Although GDF15 is linked to iron homeostasis in IDA, its increased concentrations in ACD are mostly because of inflammation.

  1,557 182 -
Prognostic significance of flow cytometric quantification of circulating endothelial cells in chronic lymphocytic leukemia
Nihal M. Heiba, Deena Samir Eissa, Shereen A. El-Shazly
January 2013, 38(1):7-12
DOI:10.7123/01.EJH.0000423009.85761.ea   
Background

Accumulating evidences have supported the role of angiogenesis in the pathogenesis and progression of chronic lymphocytic leukemia (CLL). Detection of peripheral blood circulating endothelial cells (CECs) by flow cytometry is proposed to be a noninvasive indirect marker of angiogenesis. This work aimed to quantify CECs and endothelial progenitor cells (EPCs) by flow cytometry in patients with newly diagnosed CLL compared with healthy individuals, study their relationship with established risk predictors of the disease, and assess their prognostic significance.

Materials and methods

Flow cytometric quantification of CECs and EPCs was carried out for 50 newly diagnosed B-CLL patients and 20 healthy controls. Patients were followed up for assessment of the time to first treatment, response to therapy, and disease outcome.

Results

Patients with CLL had higher counts of CECs (median, 24.6×106/l) and EPCs (median, 22.7×106/l) compared with the controls (median, 2.8 and 1.9×106/l for CECs and EPCs, respectively; P<0.001). CLL patients were subdivided according to the median values of CECs and EPCs into high and low CEC and EPC subgroups. Although high levels of CECs and EPCs were not related to established risk predictors of CLL (P>0.05), they were significantly related to higher white blood cell counts (P<0.001), shorter time to first treatment, and poor response to therapy (P<0.05).

Conclusion

This study shows that flow cytometric detection of peripheral blood CECs is a feasible indicator of abnormal angiogenesis in CLL that might be utilized as a biologic prognostic marker of a more aggressive disease course with a poor clinical outcome.

  1,137 110 -