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   Table of Contents - Current issue
April-June 2017
Volume 42 | Issue 2
Page Nos. 45-79

Online since Friday, October 6, 2017

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Development of ex-vivo expanded megakaryocyte progenitors for platelet recovery p. 45
Abdelaziz F Naglaa, Elwazir M Yasser, Hassan M Amany, Dessouky F Omar, Kamal A Hagar
Context Megakaryocytes (MKs) can be produced using umbilical cord mononuclear cells and CD34 cells in the same cocktail media. Aim The aim of this study was to produce human MKs and platelets through ex-vivo culturing of human cord blood. Patients and methods Cord blood samples were collected under sterile conditions, separated using Ficoll–Hypaque technique for mononuclear separation, followed by magnetic separation for CD34+ cells; both types of cells were cultured in the same cocktail media and followed up for 14 days. This prospective ex-vivo study was conducted to determine the differentiation potential of haematopoietic stem cells isolated from cord blood towards megakeryopoiesis lineage. Cord blood was collected in the obstetric emergency room of Suez Canal University Hospital. Stem cell separation and induction of megakeryopoiesis were carried out in the Tissue Culture Unit, Physiology Department, Faculty of Medicine, Suez Canal University. Results Both types of cells showed success towards megakaryopoiesis; however, mononuclear cells showed better results compared with CD34 cells. Conclusion Umbilical cord stem cell culture could be a new easy method for ex-vivo MK progenitor production and final platelet recovery. This success would encourage further studies in ex-vivo megakeryopoiesis and in using it in different medical aspects.
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Serum visfatin in sickle cell disease: association with frequency of vaso-occlusive crises p. 52
Deena M.M. Habashy, Marwa A Shams
Background Hypercoagulability in sickle cell disease (SCD) is multifactorial, involving abnormalities in platelet function, thrombin generation, fibrinolysis, and other multiple mechanisms. Visfatin, an adipocytokine with proinflammatory potential, can have a negative impact on the vascular endothelium in SCD. Objective We aimed to evaluate the association between serum visfatin level in SCD patients and the frequency of vaso-occlusive crises (VOC)/year in those patients. Patients and methods Sixteen sickle cell anemia (SCA) and 14 sickle β-plus thalassemia pediatric patients were studied in steady state. Twenty age-matched and sex-matched healthy individuals, who served as the control group, were evaluated with respect to serum visfatin levels by means of enzyme-linked immunosorbent assay. Results Hemoglobin (Hb) level was higher in the control group than in the SCA group (P<0.001), whereas total leukocyte count and serum visfatin were higher in the SCA group than in the control group (P=0.02 and <0.001, respectively). Hb level and platelet count were higher in the control group than in the sickle β thalassemia group (P<0.001 and 0.04, respectively). Serum visfatin was higher in the sickle β thalassemia group than in the control group (P<0.001). HbS%, serum visfatin, and frequency of VOC/year were higher in the SCA group than in the sickle β thalassemia group (P=0.002, <0.001, and 0.002, respectively). Serum visfatin was positively correlated with total leukocyte count, serum ferritin level, and frequency of VOC/year (P=0.005, 0.01 and 0.03, respectively) in the SCA group. Conclusion Serum visfatin is increased in SCD patients compared with that in healthy children and is associated with the frequency of VOC; it can be used as a predictive index for VOC occurrence and follow-up in those patients.
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Risk for red cell immunization among thalassemic patients p. 58
Eman R Saifeldeen, Mohamed A Awad, Youssef A El-Tonbary, Doaa A Aladle, Doaa M Elghannam
Background Thalassemia major is a common health problem in Egypt. However, limited data are available on the frequency of red blood cell (RBC) alloimmunization and autoimmunization in transfusion-dependent thalassemia patients. Objectives The primary aim of the current study was to determine the prevalence of alloimmunization and autoimmunization in multitransfused thalassemic patients and to assess potential clinical factors associated with RBC antibody formation. Patients and methods Sixty-five multitransfused thalassemic patients (33 male and 32 female; mean age, 9 years; range 2–37 years) who attended Mansoura University Hospital for Children were included in our study. Antibody screening and identification were carried out with the ID card microtyping system and three-cell and 11-cell panels were used (Serascan Diana 3 and Identisera Diana), respectively. Statistical analysis Statistical analysis was performed using excel program (Microsoft Office, 2010) and statistical package for social science program (SPSS Inc., Chicago, Illinois, USA), version 20. Qualitative data were presented as frequency and percentage. The χ2 and Fisher exact tests were used to compare groups. Quantitative data were presented as mean and SD or median and range. The Kolmogrov–Smirnov test was used for testing normality. For comparison between two groups, Student’s t-test or the Mann–Whitney (for nonparametric) were used. Logistic regression analysis was applied for the prediction of development of antibodies (P is significant if ≤0.05 at confidence interval 95%). Results Of 65 patients, 15 cases (23.1%) had been alloimmunized. The majority of alloantibodies were directed against the Kell (12.3%) and Rh systems (6.2%). RBC autoantibodies developed in 9.2% of patients. Only 6.2% of patients had simultaneous alloimmunization and autoimmunization. We found a significant association between RBC immunization and age at first transfusion (P=0.02), splenectomy (P<0.001), and presence of alloimmunization and autoimmunization (P=0.022). Conclusion Red cell alloimmunization is an important risk in thalassemia patients. Most alloantibodies were of the anti-K and the anti-Rh type. Extended phenotype matched blood transfusion for Rh and Kell antigens needs to be explored for preventing alloimmunization in thalassemia patients.
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Candidate markers for thromboembolic complications in adult Egyptian patients with β-thalassemia p. 64
Hadeer A Abbassy, Omar M Ghallab
Background Increased risk for thromboembolic complications in patients with thalassemia major has been attributed to the chronic hypercoagulable state. However, the underlying mechanisms are multifactorial. Objectives To investigate the candidate markers of possible tendency to thrombophilia in adult patients with thalassemia major. Methods Genetic polymorphisms of prothrombotic factors have been analyzed in 30 splenectomized patients (group I) and 30 nonsplenectomized patients (group II) as well as 30 age matched and sex matched healthy controls. In addition to routine hematological and coagulation parameters, estimation of protein C and protein S free antigen, antithrombin III activity, factor VIII, and factor IX levels was done. Plasma soluble endothelial protein C receptor levels were measured by ELISA, and factor V Leiden1691G-A, factor II polymorphism (G20210A), and methylenetetrahydrofolate reductase mutation (C677T) were detected by real-time PCR. Results A significant increase of soluble endothelial protein C receptor was observed in patients (in group I more than group II), whereas protein C and protein S were significantly reduced together with more frequent congenital thrombophilic mutations. Conclusion With the prolonged life expectancy of patients with β-thalassemia, more caution should be considered toward thromboembolic complications. Thus, after the first thrombotic event or even when they are exposed to thrombotic risk factors, they should be investigated for congenital thrombophilia, and prophylactic antithrombotic agents may be recommended.
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Effect of hydroxyurea on clinical and laboratory parameters of sickle cell anaemia patients in North–West Nigeria p. 70
Abdulaziz Hassan, Sani Awwalu, Lucky Okpetu, Aliyu D Waziri
Background Patients with sickle cell anaemia (SCA) are routinely managed with folic acid and paludrine in Nigeria. However, since the licensing of hydroxyurea (HU) by Food and Drug Administration in the USA in 1998, there has been a gradual but cautious use of HU in Nigeria, especially for SCA patients with severe disease. Aim The aim of this study was to determine the effects of HU on the frequency of hospital admissions, blood transfusion and haematological parameters among SCA patients in Zaria, Nigeria. Materials and methods This was a retrospective analytic study of patients with SCA on HU therapy at the haematology clinic of a teaching hospital in North–West Nigeria. The frequency of hospital admissions, blood units transfused per annum, haematocrit level and white blood cell (WBC) and platelet counts before and 1 year after commencing HU treatment were compared using paired sample t-test. P value lower to 0.05 was considered significant. Results In total, 18 of 689 (2.6%) registered SCA patients were on HU, and the mean age of patients was 26.5 years. The median (interquartile range) pre-HU and post-HU frequency of hospital admissions and units of blood transfused yearly were 4.5 (6) versus 1 (6) (P=0.003) and 2.5 (1) versus 0 (2) (P=0.001), respectively. Pre-HU and post-HU mean±SD haemocrit (%), WBC (×109/l) and platelet (×109/l) counts were 22.6 versus 26.6% (P<0.001), 14.8 versus 9.8 (P=0.002) and 468.3 versus 360.2 (P=0.144), respectively. Conclusion HU significantly reduced the frequency of hospital admissions and units of blood transfused with increase in haematocrit and reduction in WBC and platelet counts in patients with SCA. HU should be encouraged in patients with severe disease.
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Cytogenetic and molecular response of EUTOS risk groups in children with chronic myeloid leukemia chronic phase on imatinib treatment p. 74
Ranga R Ganta, Srividya Nasaka, Sadashivudu Gundeti, Vijay G Linga, Lakshmi S Maddali, Raghunadha R Digumarti
Introduction Recently (2011), the European Treatment and Outcome Study (EUTOS) scoring system was formulated to risk-stratify chronic myeloid leukemia (CML) chronic phase patients on imatinib treatment. CML during childhood is rare, making prognostication and management difficult. This study was conducted to investigate the cytogenetic and molecular response rates among EUTOS risk groups in pediatric CML patients on treatment with imatinib. Patients and methods Hospital records of pediatric CML chronic phase patients (age18 years) from 2009 to 2012 were analyzed retrospectively for demographic data, imatinib dose, date of initiation of treatment, EUTOS risk score, follow-up details, and cytogenetic and molecular response. Results A total of 57 children were included in this study, with a median age of 14.5 years (range: 5–18 years) and male preponderance (M:F, 1.19:1). The majority (67%) of them were in the low-risk group as per the EUTOS score and the remaining were in the high-risk group. Complete hematological response at 3 months was 100% in the low-risk and 94% in the high-risk group. Complete cytogenetic response at 12 months was 76 and 73% in the low-risk and high-risk groups, respectively. Major molecular response at 18 months was 85 and 79% in the low-risk and high-risk groups, respectively. Two-year event-free survival was 89 and 80% in the low-risk and high-risk groups, respectively. Conclusion The EUTOS low-risk cohort responded better than the high-risk cohort, but the difference was not statistically significant. Both groups had similar outcome at the end of 2 years. The EUTOS score did not predict the response and outcome in pediatric CML.
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Bone marrow granuloma due to toxoplasmosis presenting as isolated thrombocytopenia in an immunocompetent patient p. 78
Mansoor C Abdulla, Alungal Jemshad
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