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   2014| January-March  | Volume 39 | Issue 1  
    Online since January 29, 2014

 
 
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ORIGINAL ARTICLES
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.
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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.
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ORIGINAL ARTICLES
Fas (CD95) expression as a prognostic marker in acute leukemia and blastic transformation phase in chronic myeloid leukemia
Nahla A Nosair, Enaam S Abd EL Bar, Atef M Taha, Amr M Gawaly
January-March 2014, 39(1):25-31
DOI:10.4103/1110-1067.124843  
Introduction Fas is expressed on a majority of human leukemic cells. Fas-mediated cell death is involved in drug-induced apoptosis in various cell types. Hence, failure of apoptosis could lead to chemoresistance and may therefore have an impact on clinical outcome. The aim of the present study was to evaluate the percentage of Fas receptor expression on blast cells in patients with adult acute leukemia and blastic transformation phase of chronic myeloid leukemia (CML-BT), and to find out the impact of Fas expression on prognosis. Subjects and methods The participants of this study were 80 adult acute leukemia patients classified as follows: 40 acute myeloid leukemia (AML) patients, 32 ALL patients, and eight CML-BT patients. In addition, 10 age-matched and sex-matched healthy controls were also included in the study. Patients with acute leukemia were studied at diagnosis and after treatment. The diagnosis of AML, ALL, and CML-BT was assessed by morphological study, cytochemical analysis, and immunophenotyping of peripheral blood (PB) and bone marrow aspirate according to FAB classification. Fas expression on blast cells from bone marrow aspirate or PB samples of the patients or on normal monocytes, granulocytes, and lymphocytes obtained from PB samples of controls was measured using flow cytometry. The correlation between prognostic markers (age, sex, total leukocytic count, serum lactate dehydrogenase (LDH), and cytogenetic risk categories) and Fas expression levels on blast cells of leukemic patients at diagnosis was ascertained. After treatment, patients were followed up for periods ranging from 25 to 31 months. Results Fas expression was seen to be the highest on control monocytes (31.2 ± 6.95%), followed by granulocytes (24.8 ± 7.61%), whereas lymphocytes expressed the lowest levels (17.1 ± 4.01%) with a highly statistically significant difference (P < 0.001) between variables. The mean value of Fas expression by blast cells from AML patients at diagnosis was 41.72 ± 10.3%. AML patients were divided into the Fas-positive group [29 patients (72.5%)] and the Fas-negative group [11 patients (27.5%)]. The mean values of expression increased significantly from M1 to M5 with the weakest expression in M1 (20.28 ± 5.3%) and the strongest in M5 (52.91 ± 11.3%) with highly significant differences (P < 0.001) between Fas expression levels in different FAB subtypes of AML. The mean value of Fas expression in ALL patients was 42.87 ± 11.5%. Twenty-three (71.88%) patients were positive for Fas expression, whereas nine (28.12%) were negative. Fas expression was positive in 12/19 (63.2%) precursor B-ALL patients and in 11/13 (84.6%) T-ALL patients. The mean value of Fas expression was significantly higher (P = 0.039) in T-ALL (55.15 ± 7.8%) in comparison with precursor B-ALL (34.47 ± 5.76%). CML-BT patients were all transformed to AML type and expressed Fas with a mean of 31.0 ± 7.63%. Only three of them (37.5%) were Fas negative and the remaining five (62.5%) were Fas positive. The percentage of Fas-positive patients was 0% (0/1) in M1, 50% (2/4) in M2, 100% (1/1) in M4, and 100% (2/2) in M5. No significant differences could be detected between Fas-positive and Fas-negative patients with respect to baseline patient characters (age, sex, total leukocytic count, serum LDH levels, and cytogenetic risk category). Highly significant correlation was found between Fas-positive expression and response to therapy (P < 0.001) in AML and ALL patients. Fas-negative patients in CML-BT were nonresponders to chemotherapy with a significant correlation between low Fas expression and failure to respond to therapy (P = 0.049). Overall survival between Fas-positive and Fas-negative patients was statistically significant in AML, ALL, and CML-BT patients (P = 0.001, 0.002, and 0.002, respectively). Conclusion The mean value of Fas expression by blast cells from AML, ALL, and CML-BT patients at diagnosis was 41.72 ± 10.3, 42.87 ± 11.5, and 31 ± 7.63%, respectively. We can conclude that Fas receptor expression on blast cells from ALL, AML, and CML-BT patients could serve as an independent prognostic factor and help in monitoring the outcome of chemotherapy.
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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.
  1,629 131 -
Spectrum and outcome of autoimmune hemolytic anemia in children: single-center experience in 10 years
Azza A.G Tantawy, Mohammed M Al-Tawil
January-March 2014, 39(1):20-24
DOI:10.4103/1110-1067.124840  
Background Autoimmune hemolytic anemia (AIHA) is a rare disease in children. Little is known about its spectrum and outcome. The objective of this study was to analyze the clinical and hematological presentation, treatment response, and disease outcome in a cohort of children and adolescents whose initial presentation was acute AIHA. Patients and methods This was a retrospective analysis of children who presented with acute AIHA over 10 years. These cases presented to the Hematology Department, Children Hospital, Ain Shams University, Cairo, Egypt, from 2002 to 2012. Thirty-two children and adolescents were assessed for initial presentation, treatment received, and its response and disease outcome. Results In all, 78% of the patients were females. All presented with pallor, 81% with jaundice, and 34% with concomitant acute nonspecific febrile illness. Positive serology against the Epstein-Barr virus, cytomegalovirus, or mycoplasma was shown by 37.5% of the patients. Hepatosplenomegaly on initial presentation or evolving during the first year occurred in 53% of the patients. The direct antiglobulin test was negative in 6% and reticulocytopenia occurred in 12% of the patients. Ninety-four percent of patients received corticosteroids as initial therapy and 28% needed additional treatment subsequently. Complete remission (CR) at 1 month was achieved in 46.9% of patients and was highly correlated with disease outcome at 3, 12 months, and at last follow-up. Evans' syndrome was diagnosed in 50% of cases and was significantly associated with a lower subsequent CR rate. Conclusion CR of AIHA at 1 month was predictive of subsequent CR. Concordance with Evans' syndrome occurred in half of the cases with less favorable outcome.
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The value of anti-Pax-5 immunostaining in pediatric acute leukemia
Manal A Shams El Din El Telbany, Yasmin N ElSakhawy, Mahmoud T Sallam
January-March 2014, 39(1):32-35
DOI:10.4103/1110-1067.124844  
Background Pax-5 protein is a transcription factor expressed in the B-lymphoid lineage from the earliest detectable precursor to the mature B-cell stage. Its expression is closely correlated with B-cell precursor acute lymphoblastic leukemia (pre-B-ALL), which is one of the most common childhood malignancies. Aim of the work To evaluate the expression of anti-Pax-5 monoclonal antibody on leukemic blasts in pediatric patients with acute leukemia and to compare it with anti-CD20 to show its diagnostic utility. Patients and methods This study was carried out on 35 pediatric patients diagnosed with acute leukemia, who were divided into two groups: Pre-B-ALL and Acute, myeloid leukemia (AML). All patients were subjected to a complete assessment of history, full clinical and laboratory examination, bone marrow aspirate and biopsy, and immunophenotyping. Immunostaining of bone marrow trephine biopsy specimens using Pax-5 and CD20 monoclonals was carried out for all cases. Results Pax-5 was positive in 79.1% of cases in the pre-B-ALL group and negative in all AML patients. CD20 was positive in 37.5 and 18% of cases in the ALL and the AML groups, respectively. Pax-5 appeared to be more sensitive (79.2%) and more specific (100%) than CD20 in differentiating ALL and AML. Conclusion Our results highlight the beneficial role of including Pax-5 in an immunohistochemical panel to diagnose acute leukemia of B-cell lineage.
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