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ORIGINAL ARTICLES
Blood indices to differentiate between β-thalassemia trait and iron deficiency anemia in adult healthy Egyptian blood donors
AR Soliman, G Kamal, A Elsalakawy Walaa, TH Sallam Mohamed
July-September 2014, 39(3):91-97
DOI:10.4103/1110-1067.148223  
Background β-Thalassemia trait (BTT) often shows microcytosis, a normal or an increased red blood cell (RBC) count, and an elevated level of HbA 2 , which provide the basis for laboratory screening. BTT is an important differential diagnosis of iron deficiency anemia (IDA). Donors with BTT have hemoglobin values comparable with normal; hence, they are accepted for donation and they usually escape diagnosis. Aim The aim of this work was to differentiate BTT from IDA through blood indices performed in routine complete blood count. Patients and methods A total of 200 samples were obtained from apparently healthy adult Egyptian blood donors randomly. Complete blood count and mean corpuscular volume (MCV) were performed to all individuals. Hemoglobin electrophoresis and/or serum ferritin was performed to samples with MCV less than 78 fl. Results Prevalence of BTT in this study was 6%, whereas IDA represented 4.5% of total 200 samples investigated. The cutoff value of MCV 73 fl was 91.7% sensitive and 100% specific in differentiating BTT from IDA. Red blood cell distribution width at level 14.5% or below can differentiate BTT from IDA with 83.3% sensitivity and 100% specificity. RBCs count at value above 5.47 million/mm 3 can differentiate BTT from IDA with 100% sensitivity and 100% specificity. Conclusion The cutoff values of MCV 73 fl or less, RBC count above 5 × 10 6 /mm 3 , and red blood cell distribution width 14.5% or less were suggested to be associated with a high probability of BTT.
  5,483 446 -
Is iron treatment related to weight gain in female patients with iron deficiency anemia?
Osman Yokus, Habip Gedik
April-June 2016, 41(2):42-44
DOI:10.4103/1110-1067.186395  
Objectives To assess the relationship between iron treatments and to gain weight in the female patients. Materials and methods This study was conducted prospectively between 2013 and 2014 to assess the relationship between iron treatments and to gain weight in the female patients who were admitted to hematology clinic, Ministry of Health İstanbul Research Hospital with a diagnosis of iron deficiency anemia and iron therapy between the ages of 18 and 50 and signed informed consent. Results After 3 months of iron therapy, significant hemoglobin (Hgb) increase was observed in 30 of 33 patients. Weight gain and Hgb increase have not been found in three patients. Hgb increases in patients who gained weight more than 3 kg were found to be statistically significant than patients who gained weight 3 kg or less (P = 0.008). Conclusion Obesity and metabolic syndrome are current health problems in terms of mortality and morbidity. Weight gain related to iron therapy is a common problem in female patients with iron deficiency anemia. Patients under iron therapy should be counseled in terms of weight gain complication and benefits of diet and followed up serum ferritin and Hgb levels to prevent prolonged iron therapy. Physicians should pay attention to patients who do not gain weight without dieting.
  5,357 118 -
Platelet indices: consideration in thrombocytopenia
DA Elsewefy, BA Farweez, RR Ibrahim
July-September 2014, 39(3):134-138
DOI:10.4103/1110-1067.148240  
Background There is increasing evidence that platelet indices, such as mean platelet volume (MPV), platelet distribution width (PDW), and platelet large cell ratio (P-LCR), have a significant role in the discrimination between hyperdestructive thrombocytopenia and hypoproductive thrombocytopenia, and they can be of great help as they are routinely generated by automated cell counters. Objective In this study, we aimed to assess the sensitivity and specificity of these indices and set cutoff values that aid in the diagnosis of thrombocytopenia cause. Materials and methods We recruited 20 individuals as the control group and 80 thrombocytopenic patients, who were divided into two groups: group I ( n = 40) included newly diagnosed immune thrombocytopenic purpura (ITP) patients (hyperdestructive thrombocytopenia), whereas group II ( n = 40) included hypoproductive thrombocytopenia patients. The MPV and platelet distribution width were derived from automated cell counter results. The P-LCR was calculated. Results In ITP (hyperdestructive thrombocytopenia) patients, the MPV and P-LCR were significantly higher; the best cutoff value for MPV was greater than 9.7 fl and for P-LCR was greater than 33.6%, with a diagnostic accuracy of 70.1 and 99.6%, respectively. Conclusion The MPV and P-LCR provide information about the underlying conditions of thrombocytopenia. These indices should be considered in the diagnosis of thrombocytopenia. The P-LCR can be safely relied upon for a positive diagnosis of ITP.
  4,844 412 -
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.
  1,993 246 1
Inducing hepatogenic differentiation of human mesenchymal stem cells derived from umbilical cord blood
Enas A Elzamarany, Nahla A Nosair, Dareen A Mohamed, Moustafa Z Moustafa, Ghada M Balah
January-March 2014, 39(1):6-12
DOI:10.4103/1110-1067.124838  
Background and aim The present work aimed to study the ability of human mesenchymal stem cells (MSCs) derived from umbilical cord blood (UCB) to transdifferentiate into hepatocytes and to assess the characterization of the transformed cells in vitro. Materials and methods MSCs were isolated from 30 UCB samples collected aseptically from completely separated placentas of full-term deliveries. The separation of MSCs was carried out from freshly isolated mononuclear cells suspensions in a primary culture for 2 weeks. MSCs were identified before induction by cytochemical stain for periodic acid-Schiff and morphology. Then, they were induced to transdifferentiate into hepatocytes by hepatogenic medium, and differentiation of hepatocytes was confirmed by morphological and functional assessments of urea production, glycogen storage, and immunocytochemistry of α-fetoprotein. Results We successfully isolated 12 MSCs units from 30 full-term UCB samples (40%). The cells showed positive staining for periodic acid-Schiff, indicating that they retain the characteristics of MSCs. The response of UCB-derived MSCs to hepatogenic medium containing hepatocyte growth factor was assessed by changes in the morphology that occurred within 2 weeks in most of the cells. The morphological changes were observed closely and the induction process was monitored by immunocytochemistry for α-fetoprotein, urea production, and glycogen storage. The results were positive in most of the induced hepatocytes, with some variations from sample to sample because of the variability in the cell count in each UCB unit used. Conclusion We concluded that MSCs in human UCB can differentiate into viable functioning hepatocytes when cultured in hepatogenic conditioned medium in vitro.
  1,753 137 -
Lipid profile in children with β-thalassemia major
Samera Z. Sayed, Sheren E. Maher, Ghada Adel, Lamea Hamdy
October 2012, 37(4):183-186
DOI:10.7123/01.EJH.0000418696.53110.08   
Background

β-Thalassemia major is a very serious blood condition, as affected patients are unable to synthesize enough healthy red blood cells and depend on blood transfusions throughout their life.

Aim of work

The aim of the study was to evaluate the lipid profile in patients with β-thalassemia major.

Patients and methods

Fifty patients with β-thalassemia major and 25 healthy controls were included in this study. They were subjected to complete history taking, a thorough clinical examination, and laboratory investigations including complete blood count, liver function test, and assessment of serum ferritin levels and fasting lipid profile including total cholesterol, high-density lipoprotein (HDL), low-density lipoprotein (LDL), and triglycerides (TGs) levels.

Results

Patients with β-thalassemia major showed significantly lower total cholesterol, HDL-cholesterol, and LDL-cholesterol when compared with controls. Serum TG levels of β-thalassemia major patients were found to be significantly higher than the levels in control individuals. Our results revealed that the lipid profile changed in patients with thalassemia major.

Conclusion

In thalassemic patients, adequate chelation therapy with normalization of serum ferritin level and monitoring of TGs is highly recommended, and they are treated aggressively if the levels are increased. Several interventions including antioxidant therapy and vitamin-lowering and lipid-lowering agents should be used in high-risk patients with β-thalassemia major to decrease the risk of atherosclerosis.

  734 1,116 -
REVIEW ARTICLE
Nanotechnology-based artificial platelets
Feroz Alam, Mohammed Naim, Suhail-ur Rahman, Mariam Shadan
January-March 2014, 39(1):1-5
DOI:10.4103/1110-1067.124836  
Platelets are essential components of blood. Because of drawbacks in collection and storage of platelets, marked shortage is felt especially in the areas endemic for dengue fever and malaria. To meet the shortage, various synthetic and semisynthetic compounds are being produced and tested worldwide, which either enhance platelet function or are a substitute to it. In this study, we discuss these semiartificial and artificial substitutes of platelets along with other compounds that enhance the activity of platelets, with special emphasis on their mechanism of action and biofeasibility.
  1,596 184 -
ORIGINAL ARTICLES
Zinc and malondialdehyde levels among Egyptian patients with acute myeloid leukemia and their relation with disease phenotype and genotype
Inas A Asfour, Maryse S Ayoub, Amro M.S. El-Ghammaz, Ibtesam M Khalifa
October-December 2015, 40(4):153-158
DOI:10.4103/1110-1067.170190  
Objectives This study was conducted to evaluate serum zinc and plasma malondialdehyde (MDA) levels in de-novo acute myeloid leukemia (AML) before and after induction chemotherapy and their relation with AML phenotype and genotype. Materials and methods Twenty-five AML patients were subjected to serum zinc evaluation using flame atomic absorption spectrophotometry and plasma MDA evaluation using colorimetric method at day 1 before induction chemotherapy and at day 21 after induction chemotherapy. Results Pretreatment MDA levels were higher in patients in comparison with controls (P = 0.03). Pretreatment zinc levels differed significantly compared with post-treatment levels (P = 0.005). The percentage of bone marrow infiltration by blasts at diagnosis correlated inversely with zinc levels (P = 0.011) and positively with MDA levels (P = 0.041). Finally, pretreatment MDA levels were higher among patients harboring adverse cytogenetics (P = 0.004). Conclusion The elevated plasma MDA status at diagnosis in AML patients correlates with a higher tumor burden in the bone marrow and adverse cytogenetic risk.
  487 1,123 -
Plasma interleukin-22 and its cellular receptor (IL-22RA1) expression in chronic lymphocytic leukemia
Nihal M. Heiba, Shereen A. Elshazly
November 2013, 38(4):123-129
DOI:10.7123/01.EJH.0000434281.48881.82   
Background

Chronic lymphocytic leukemia (CLL) is an environment-dependent hematologic malignancy where interactions with accessory cells through cytokines and their receptors seem to confer a survival advantage, thus contributing to disease progression. Interleukin-22 (IL-22) is a T-cell-derived cytokine that promotes cell proliferation and survival through interaction with its receptor IL-22RA1, normally absent in normal immune cells, including B and T lymphocytes.

Aim

This study aimed to determine the plasma levels of IL-22 and the expression of IL-22RA1 on malignant cells in patients with B-cell CLL (B-CLL), together with their relation to clinical and prognostic characteristics of the disease.

Patients and methods

The study was carried out on 62 newly diagnosed B-CLL patients. Twenty-five age-matched and sex-matched healthy individuals served as controls. Patients were diagnosed, according to the International Workshop on CLL guidelines, by cytomophology, immunophenotyping, conventional cytogenetic analysis, and fluorescence in-situ hybridization. Plasma IL-22 levels were measured by an enzyme-linked immunosorbent assay and the expression of IL-22RA1 on leukemic cells was assessed by flow cytometry.

Results

Plasma IL-22 was significantly higher in B-CLL patients (range, undetectable 62.9 pg/ml; median, 6.6) compared with control participants (range, undetectable 6.4 pg/ml; median, undetectable) (P<0.01). IL-22RA1 expression was negative in all normal controls, whereas in B-CLL patients it was positively expressed in 35/62 CLL cases (56%). Taking the median level of IL-22RA1 expression in CLL patients as a cutoff level, overexpression (≥10%) was observed in 32/62 (52%) cases. IL-22RA1 expression correlated significantly positively with plasma levels of IL-22 (rs=0.817; P<0.01). Patients presenting with high CD38 expression had significantly higher plasma IL-22 levels compared with those with low CD38 (undetectable 62.9 pg/ml; median, 19.3 vs. undetectable 50.1 pg/ml; median, 3.1) (P<0.01) as well as overexpression of IL-22RA1. No significant relation could be established between either plasma IL-22 levels or IL-22RA1 expression with other clinical features or prognostic criteria of CLL.

Conclusion

This is the first report to describe the aberrant expression of the IL-22 signaling pathway in B-CLL and to link its overexpression with high CD38 expression, a known poor prognostic marker of the disease.

  483 1,077 -
Contributions of flow cytometry in the evaluation of myelodysplastic syndrome patients
Amany H. Mansour, Eman Kassm, Tawfik Elkhodary, Samar Taha
November 2013, 38(4):141-148
DOI:10.7123/01.EJH.0000434284.33634.ca   
Background

Myelodysplastic syndromes (MDS) are a group of malignant myeloid hematopoietic disorders. The diagnosis of MDS can be difficult, especially in cases with a low blast count and a normal karyotype. Flow cytometry has been used to distinguish MDS from nonclonal cytopenias. No one single simple flow cytometric parameter has been proposed to be diagnostic of MDS.

Aim

The aim of the present study was to evaluate immunophenotypic alterations in typical MDS patients and whether these abnormalities help in the differentiation process between MDS with nonclonal disorder and leukemic patients.

Materials and methods

Marrow aspirates from 29 patients, including 13 with MDS, 16 with acute myeloid leukemia, and 18 with nonclonal disorders (normal controls), were examined in this study. Their immunophenotypes were analyzed using flow cytometry. Blasts, nonblast myeloid cells, and monocytes were gated on the basis of CD45 expression and side scatter (SSC).

Results

Comparison among the three groups showed that the granulocytic lineages of MDS showed decreased SSC compared with the controls (P<0.005 and P<0.000, respectively), altered CD45 intensity (P<0.004), decreased CD10-positive granulocytes (P<0.02), and a higher CD56 positive expression in the MDS and leukemic group (P<0.05 and P<0.001, respectively). Also, decreased intensity of CD11b (P<0.03) was observed in the MDS group. The expression rate of CD123+ was significantly higher in MDS patients than that in normal controls (P<0.0001).

Conclusion

Gating of the granulocytic region is a relatively easy method for MDS immunophenotyping. Among the parameters studied, SSC, CD10, CD123, and CD56 were the most useful for differentiating MDS from nonclonal disorders, whereas immunophenotypic changes in MDS appear to be useful for differentiating MDS from nonclonal disorders.

  1,349 128 -
Rapid detection of multiple β-globin gene mutations by a real-time polymerase chain reaction in β-thalassemia carriers
Salwa M. Youssef, Mohsen S. El Alfy, Amany A. Osman, Dina A. Khattab, Mervat A. El Feky, Marwa E. Hussein
July 2012, 37(3):147-155
DOI:10.7123/01.EJH.0000416544.53925.2a   
Background

β-Thalassemia is a heterogeneous disorder caused by mutations that reduce or abolish the synthesis of the β-globin chain. The clinical severity of thalassemia major makes it a priority genetic disease for prevention programs involving population screening of heterozygotes and an optional prenatal diagnosis for carrier couples.

Aim of the study

This study aimed to determine the most common β-globin gene mutations in Egypt using a real-time PCR and fluorescently labeled hybridization probes specific for each mutation and to assess the feasibility of introducing this technique in an overall thalassemia prevention program.

Participants and methods

The study was carried out on 45 individuals: 37 β-thalassemia carriers [including five amniotic fluid (AF) samples], seven β-thalassemia major cases (including two AF samples), and one normal AF sample. The most common β-thalassemia mutations were characterized by real-time PCR with fluorescently labeled hybridization probes specific for IVSI-110, IVSI-1, IVSI-6, codon 37, and codon 39 in 28/37 (75.7%) carriers.

Results

The most common mutation encountered was IVSI-110 (46%), followed by IVSI-1 (16.2%) and then IVSI-6 (13.5%). Codon 37 and codon 39 were not characterized in any sample. The genotype of the uncharacterized carriers was determined using a less sensitive method (reverse hybridization technique) and a relatively less common set of mutation was characterized as follows: IVSII-1(10.8%), codon 5 (5.4%), IVSII-745 (5.4%), and IVSI-116 (2.7%). The overall number of alleles detected using both techniques was calculated to be 51. The real-time PCR alone, with its assigned probes, detected 38/51(74.5%). Thirteen mutations (13/51=25.5%) remained uncharacterized by this technique (because of the unavailability of the corresponding probes). However, the reverse hybridization technique detected 48/51 alleles (94.1%). However, comparison between both techniques in terms of the shared mutations showed that the real-time PCR detected 38/38 (100%) of these mutations, whereas the reverse hybridization technique detected only 36/38 (94.7%).

Conclusion

Real-time PCR is a very rapid and accurate method for the detection of the β-thalassemia mutation, which may be valuable in cases for which a rapid decision has to be taken. Impediments to prenatal diagnosis as encountered in this study were attributed to refusal of termination of pregnancy by the family for religious/reasons, abortion following amniocentesis, and failure to determine the correct genotype of the AF analyzed.

  1,295 174 -
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.
  1,316 150 -
Novel immunophenotyping marker in acute myeloid leukemia: does it implicate prognosis?
Amira Y Abd El-Naby, Amr Gawaly, Shereen Awni
October-December 2016, 41(4):155-160
DOI:10.4103/1110-1067.198646  
Introduction AML is a heterogeneous group of neoplasms that affect hematopoietic cells responsible for the production of myeloid lineages in bone marrow (BM). Novel immunophenotyping marker in AML (CD30) is a 120 kDa cell membrane glycoprotein that shares sequence homology with tumor necrosis factor (TNF) receptors; it is expressed by myeloblasts (CD34+ and/or CD117+) in a substantial number of cases of non Monocytic AML, either de novo or arising from MDS. CD30 expression is more common in AML patients with unfavorable chromosomal abnormality. It is suggested that CD30 could be a target for therapy by using anti-CD30 antibodies in a subset of patients with non Monocytic AML; also it might provide useful information for patient prognosis and stage of disease. Aim of work Is to assess novel immunophenotyping marker in AML (CD30) and cytogenetic abnormality and its correlation with prognosis. Materials and methods This prospective cohort study included 120 patients with newly diagnosed acute myeloid leukemia. Their ages ranged from (2 to 75) years with a mean of (40.15±23.55) years. Chromosomal analysis by karyotyping was done in all AML patients and Multicolor flow-cytometry immunophenotypic analysis was performed on bone marrow aspirates of the AML patients using fluroisothiocyanate (FITC) conjugated antibodies to assess CD30 expression on BM myeloblasts. Results The previous study showed that CD30 was expressed in all AML cases rather than M4 and M5 cases that showed negative expression. In addition, this study showed that there is more CD30 expression in myeloblasts with unfavorable chromosomal abnormalities. A significant association between platelets counts and CD30 expression was also observed. There was a higher degree of thrombocytopenia and a greater tendency to have higher leucocytic counts in patients having +ve CD30 expression than those with –ve CD30 expression. Conclusion The analysis of CD30 expression has a potential role to be used as a prognostic marker in AML.
  412 997 -
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.
  1,175 187 1
Detection of Janus kinase 2 V617F mutation in healthy cigarette smokers
Samy B.M. El-Hady, Amina M. Elnaggar, Eman Almasry
July 2013, 38(3):97-101
DOI:10.7123/01.EJH.0000430746.10788.76   
Background

Janus kinases are cytoplasmic tyrosine kinases that mediate signaling from the cytokine receptors to the cell nucleus. Janus kinase 2 mutation (JAK2 V617F) analysis has been endorsed by the WHO for diagnosing polycythemia vera, essential thrombocythemia, and primary myelofibrosis. The aim of this study was to assess JAK2 V617F point mutation in healthy cigarette smokers compared with healthy nonsmokers and to correlate the presence of this mutation with some clinical and laboratory variables.

Materials and methods

Group I comprised 34 cigarette smokers who have been smoking 10 or more cigarettes per day, every day of the week, for at least 10 consecutive years. Group II comprised 42 men who were nonsmokers with no history of drug abuse. In addition to routine laboratory investigations, detection of JAK2 V617F point mutation in peripheral blood neutrophils was assessed for all participants.

Results

In this study, we found an increased percentage of JAK2 V617F mutation in cigarette smokers compared with nonsmokers. Further, we found a significant positive correlation between the percentage of JAK2 V617F mutation and age in both groups.

Conclusion

JAK2 V617F mutation has been detected in the healthy population; however, its incidence significantly increases in cigarette smokers. The mechanisms leading to excess JAK2 mutation and the importance of this mutation in smokers are yet to be elucidated and an adequate follow-up of healthy individuals who carry the mutation is recommended.

  1,202 102 -
The application of eosin maleimide-binding test in the diagnosis of hereditary spherocytosis among undiagnosed cases of chronic hemolytic anemia in children
Wessam M El Gendy, Hoda M Hassab, Amal M Ghanem, Irene M Lewis, Sarah M Nawar
July-September 2014, 39(3):109-113
DOI:10.4103/1110-1067.148229  
Background Conventional diagnosis of hereditary red blood cell (RBC) membrane disorders, in particular hereditary spherocytosis (HS), is labor intensive, time consuming and requires at least 2 ml of blood, which might be impractical in the neonatal period. Participants and methods We evaluated the use of eosin-5-maleimide (EMA) as a rapid screening test for patients with HS. RBCs from 74 healthy controls and 66 anemic children (35 HS and 31 other hemolytic anemias; 10 cases diagnosed as thalassemia, eight cases of autoimmune hemolytic anemia, one case of ovalocytosis and 12 cases of undiagnosed hemolytic anemia) were stained with EMA and analyzed for their mean fluorescence intensity using flow cytometry. Results RBCs from patients with HS showed a greater degree of reduction in mean fluorescence intensity of EMA compared with those from normal controls and patients with other hemolytic diseases. These findings showed that the fluorescence flow cytometric-based method is a simple, sensitive and reliable diagnostic test for RBC membrane disorders using a small volume of blood, and results could be obtained within 2 h. Such a method could serve as a first-line screening for the diagnosis of HS in routine hematology.
  1,132 136 -
Prognostic significance of serum monoclonal immunoglobulin in B-chronic lymphocytic leukemia
Mona A. El-Hussiny, Lamiaa Ibrahim, Emad Azmy, Sameh Shamaa
October 2012, 37(4):240-245
DOI:10.7123/01.EJH.0000419280.71909.65   
Background

Serum immunoglobulin (Ig) paraprotein can be detected in a subset of patients with chronic lymphocytic leukemia (CLL) by serum protein electrophoresis and immunofixation electrophoresis. CLL with Ig paraproteinemia had an inferior survival compared with patients with CLL without serum paraprotein.

Participants and methods

The present study was carried out on 100 patients with B-CLL (60 men and 40 women) ranging in age from 33 to 75 years. The staging of CLL was performed according to the Binet staging system. Venous blood samples were obtained from B-CLL patients for a complete blood count. Serum was separated for the measurement of lactate dehydrogenase (LDH), β2-microglobulin (β2-MG) levels by ELISA, and Ig paraprotein. Bone marrow aspiration was carried out for all B-CLL cases. Prognostic markers of CD38 and &zgr;-chain-associated protein kinase-70 (ZAP-70) expression were also analyzed.

Results

Twenty-two patients had Ig paraproteinemia of a total of 100 untreated patients with CLL, frequency 22%. There was a highly significant elevation in LDH, β2-MG, CD38, ZAP-70, IgG, and IgM in CLL with monoclonal paraprotein versus CLL without monoclonal paraprotein. There were positive correlations of serum IgG paraprotein and serum IgM paraprotein with advanced Binet stage (P<0.00 and P=0.00), high level of β2-MG (P=0.00 and 0.00) and LDH (P=0.00 and 0.00), CD38 positivity (P=0.00 and 0.02), and ZAP-70 positivity (P=0.00 and 0.01). Also, there was a positive correlation between high serum β2-MG concentration with Binet stage (P=0.00), high level of LDH (P=0.00), CD38 positivity (P=0.00), and ZAP-70 positivity (P=0.02). After a follow-up of 60 months, 15 patients (15%) died. Eight (36.3%) patients with Ig paraproteinemia died during the observation period, whereas among the 78 patients without Ig paraproteinemia, seven (8.9%) died.

Conclusion

β2-MG and Ig paraprotein serve as poor prognostic markers for B-CLL. Patients with CLL with serum Ig paraprotein represent a heterogeneous group with an inferior clinical outcome. Serum Ig paraprotein might be applied for the assessment of prognosis in patients with CLL.

  1,143 97 -
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.
  1,085 132 -
Cerebrospinal fluid and serum levels of monocyte chemoattractant protein 1 in acute leukemia patients: correlation to other prognostic factors
Mohammad I Ahmed, Dalia A Nafe, Maha Y Kamal
October-December 2014, 39(4):183-189
DOI:10.4103/1110-1067.153942  
Background Acute leukemia (AL) is a malignancy with accumulation of blasts in the bone marrow (BM). The blast cells enter into the peripheral blood stream and secondary localized extramedullary sites. The regulation of this process has not been clearly explained so far. Objective The aim of this study was to evaluate the cerebrospinal fluid (CSF) and serum levels of monocyte chemoattractant protein 1 (MCP1) in AL patients with correlation to other prognostic factors and tumor load. In a case-control study, 80 de-novo AL patients and 30 healthy age-matched controls were included. All patients were subjected to thorough history taking, full physical examination, and laboratory investigations such as complete blood count, BM examination, and flow cytometry (by enzyme-linked immunosorbent assay method). Results MCP1 was significantly elevated in both blood and CSF in AL patients in comparison with the control group (P < 0.05). In AL patients, there was a significant positive correlation between MCP1 level in both blood and CSF and the following parameters: hepatomegaly, splenomegaly, lymphadenopathy, purpura, fever, platelet count, white blood cells, and blast % in peripheral blood and BM (P < 0.05) were observed. There was no correlation between MCP1 in blood and CSF and Hb, lactate dehydrogenase and first hour of erythrocyte sedimentation rate. Conclusion Significant increase in the CSF and blood levels of MCP1 level was observed in AL patients compared with the control group. This may be related to extramedullary leukemic infiltration, tumor load, and disease activity in these patients. Egyptian J Haematol 39:-0 © 2014 The Egyptian Society of Haematology.
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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.
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REVIEW ARTICLE
How to approach drug-induced agranulocytosis in ICU
Seema Mahant, Piyu Deo Mahant, Upasana Shobhane
July-September 2015, 40(3):109-112
DOI:10.4103/1110-1067.164724  
Many drugs can cause agranulocytosis and neutropenia by bone marrow suppression. Drug-induced agranulocytosis (DIA) is a relatively rare, but life-threatening disorder that frequently occurs as an adverse reaction to drugs. The overall incidence of DIA ranges from 2.4 to 15.4 cases/million patients exposed to drugs per year. DIA remains a serious adverse event because of the occurrence of severe sepsis with severe deep infections (such as pneumonia), septicemia, and septic shock in around two thirds of patients. In this setting, older age (>65 years), septicemia or shock, metabolic disorders such as renal failure, and a neutrophil count below 0.1×10 9 /l are poor prognostic factors. The severity of neutropenia (<0.1×10 9 /l) and its duration (>10 days) may also impact negatively on the outcome. Commonly used drugs such as antibiotics (b-lactam and cotrimoxazole), antiplatelet agents (ticlopidine), antithyroid drugs, sulfasalazine, neuroleptics (clozapine), antiepileptic agents (carbamazepine), nonsteroidal anti-inflammatory agents, and dipyrone are the most common causes of neutropenia and agranulocytosis. Recent investigations suggest that there are at least three mechanisms by which it can be produced, namely, differences in drug pharmacokinetics, abnormal sensitivity of myeloid precursors, and adverse immune responses to drug administration. Genetic factors are important and could act by any of the above mechanism. In management, use of hematopoietic growth factors, such as granulocyte colony-stimulating factor and granulocyte-macrophage colony-stimulating factor, reduced mortality rate from 21.5 to 5%. Now a days, physicians use many drugs to increased life expectancy, as well as the development of new agents, should be aware of this complication and its management.
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ORIGINAL ARTICLES
Evaluation of the procoagulant potential of endothelial microparticles CD144 (VE-Cadherin) positive in coronary syndrome patients
Amany H Mansour, Azaa M Abd Elbaky, Shahir Kamal, M Noura, Abdulrahman Alshaikh
July-September 2014, 39(3):143-148
DOI:10.4103/1110-1067.148244  
Background Apoptotic microparticles are responsible for almost all tissue factor activity of the plaque lipid core. We hypothesized that elevated levels of procoagulant microparticles could also circulate in the peripheral blood of patients with recent clinical signs of plaque disruption and thrombosis. The present study included 60 acute coronary syndrome (ACS) adult patients. Group I included 30 patients with diabetes mellitus who presented with ACS. Group II included 30 nondiabetic patients complaining of ACS and 25 healthy individuals as controls. ACS patients were further classified according to laboratory and radiological findings (troponin test and ECG) as follows: group A included 16 ST-segment elevation myocardial infarction (STEMI) patients, group B included 19 non-ST-segment elevation myocardial infarction (NSTEMI) patients, and group C included 25 patients with unstable angina. Traditional laboratory investigations and special laboratory assessments of CD144 fluorescein isothiocyanate by flow cytometry were performed. Results The present study found highly elevated CD144 percentages in diabetic ACS patients compared with healthy controls (P≤0.0001(, and highly elevated creatine kinase-MB (CK-MB), fasting sugar, total cholesterol, triglyceride, HDL, and LDL (P = 0.0001, 0.0001, 0.0002, 0.0002, 0.0001, and 0.0001, respectively). In contrast, nondiabetic ACS patients had significantly elevated CD144, CK-MB, total cholesterol, triglyceride, HDL, and LDL (P = 0.0001, 0.0001, 0.0001, 0.0021, 0.0001, and 0.0021, respectively), whereas fasting sugar and HbA1c did not change significantly. However, the patients in group B (NSTEMI) had significantly elevated CD144% in comparison with patients with unstable angina (group C) ( P = 0.05), but patients with group A (STEMI) had significantly elevated CK-MB compared with patients with unstable angina (group C) (P = 0.02). Conclusion The high levels of circulating microparticles of endothelial origin are increased in diabetic patients with coronary artery disease, suggesting an important role for endothelial injury in the prediction of ACS. Hyperglycemia in ACS is associated with enhanced local thrombin generation and platelet activation, as well as unfavorably altered clot features in patients with and without a previous history of diabetes.
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The usefulness of immunoglobulin-like transcript-3 receptor expression in the diagnosis of acute myeloid leukemia with monocytic differentiation
Maha Atfy, Hoda F Ebian, Ashraf M Elhefni, Hebatallah H Atteia
July-September 2014, 39(3):122-127
DOI:10.4103/1110-1067.148235  
Background Immunoglobulin like transcript (ILT3) is an immunohibitory transmembrane receptor expressed by plasmacytoid dendritic cells (PDCs), monocytoid dendritic cells (MDCs), and monocytes/macrophages. ILT3 has been previously utilized as a highly sensitive and specific marker for the identification of aggressive chronic lymphocytic leukemia (CLL) and to distinguish AML with monocytic differentiation from other types of AML. Aim and Methods0 Therefore, in the current study, we investigated the diagnostic impact of ILT3 receptor expression in 72 Egyptian patients having AML with monocytic differentiation and 20 healthy volunteer using flow cytometry technique. Results Our results demonstrated significant overexpression of ILT3 receptor (P < 0.001) in all AML cases displaying monocytic differentiation patients. ILT3 receptor expression was positively correlated with CD4, CD14, CD64, CD11c, MPO and NSE. Conclusion These data present a significant role for ILT3 receptor expression in combination with CD14, CD64, MPO and NSE in diagnosis of AML with monocytic differentiation. Further clinical studies should be directed towards ILT3 receptor blockade as a future target for AML immunotherapy.
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Evaluation of multidrug resistance in acute leukemia using real-time polymerase chain reaction
Eman Mossad, Rania Bakry, Hosney Badrawy, Eman Ahmed Hasan, Mohamed R Khalaf
January-March 2014, 39(1):13-19
DOI:10.4103/1110-1067.124839  
Background Despite the advances in the cure rate for acute leukemia, ~25% of affected patients develop relapses. Expression of genes for the multidrug resistance (MDR1) and breast cancer-resistance protein (BCRP) may confer the phenotype of resistance to the treatment of acute leukemia. Objective To analyze the expression of the MDR1 and BCRP genes in new cases of acute leukemia using real-time PCR (RT-PCR) and to determine the correlation between their expression and overall survival (OS). Patients and methods Patients diagnosed with acute myeloblastic leukemia (AML) (n = 15) and acute lymphoblastic leukemia (ALL) (n = 35), and 20 blood donors as a control group were included in this study. The expressions of mRNA for the MDR1 and BCRP genes were assessed by RT-PCR. Myeloid surface markers such as CD34, CD33, CD13, and CD14 and lymphoid surface markers such as CD3, CD5, CD2, CD4, CD8, and CD19 were analyzed using flow cytometry. Results The groups with the MDR gene and the BCRP gene showed a highly significant difference compared with the control group (P < 0.000). The relation between MDR and BCRP in both AML and ALL groups showed no significant difference. There was a significant difference between BCRP expression in the AML and ALL groups (P < 0.01). There was no significant difference in the OS between MDR+ cases and MDR- cases in the AML and ALL groups. In contrast, the OS in BCRP+ cases and BCRP- cases showed a significant difference between AML and ALL groups (P < 0.01). No significant difference was detected between OS in AML (MDR+, CD34+) and AML (MDR+, CD34−). In contrast, OS between AML (BCRP+, CD34+) and AML (BCRP+, CD34−) showed a significant difference (P < 0.01). The difference between OS in ALL (MDR+, CD34+) and ALL (MDR+, CD34−) was not significant. In contrast, a significant difference was detected between OS in ALL (BCRP+, CD34+) and ALL (BCRP+, CD34−) (P < 0.01). OS in the AML group that was BCRP+ (CD13+) showed a significant difference (P < 0.01). In the ALL group, the association between MDR+ and CD19+ or BCRP+ and CD19+ did not affect the survival significantly. Conclusion We concluded that the evaluation of the expression of genes for resistance to antineoplastic drugs in acute leukemia upon diagnosis, and particularly the expression of the BCRP gene, may be of clinical relevance.
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Parvovirus B19, rubella, Epstein–Barr, and cytomegalovirus and idiopathic thrombocytopenia in Egyptian children, single-center study
Maysaa El Sayed Zaki, Hanaa Morcous
July 2012, 37(3):178-181
DOI:10.7123/01.EJH.0000416549.07291.e2   
Background

Idiopathic thrombocytopenic purpura (ITP) in children is usually a self-limiting disorder. It may follow a viral infection or immunization and is caused by an inappropriate response of the immune system. Many viruses, such as parvovirus B19, cytomegalovirus (CMV), Epstein–Barr virus (EBV), and rubella, are implicated in the occurrence of ITP.

Aim

The aim of the study was to investigate the occurrence of viral-associated ITP in Egyptian children.

Materials and methods

Viral studies included specific immunoglobulin M for rubella, EBV, and CMV. In addition, molecular detection for parvovirus B19 was carried out.

Results

Positive viral markers either by positive serology immunoglobulin M for rubella, CMV, or EBV or by a molecular study for parvovirus B19 were detected in 19 patients (38.8%). The most common viral infection was parvovirus B19 (30.6%), followed by EBV (16.3%), rubella (12.12%), and CMV (10.2%).

Conclusion

We conclude that in a large proportion of children with ITP in our region, an association with markers of acute viral infections similar to those of rubella, EBV, and CMV is present. Moreover, a significant proportion of the children had occult parvovirus B19 viremia. A study of occult viral infections is recommended in children with ITP.

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