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2014| January-June | Volume 3 | Issue 1
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ARTICLE
Expression of Aberrant Antigens CD7 and CD19 in Adult Acute Myeloid Leukaemia by Flow Cytometry
Sinan Y Muhsin, Subh S Al-Mudallal
January-June 2014, 3(1):1-13
Background
: Flow cytometric immunophenotyping (FCI) is an indispensable tool for quantitative and qualitative evaluation of antigen expression of hematopoietic cells. From a diagnostic and therapeutic point of view, distinction between acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL) and for diagnosis and definition of particular AML subtypes. Aberrant antigen expression (i.e. expression of an antigen which is inappropriate for a lineage) is rarely seen in normal bone marrow cells and their incidence is varied between various studies.
Objectives :
To explore the existence of the aberrant lymphoid antigens (CD7) and (CD19) in adult acute myeloid leukemia patients and their distribution among French-American-British (FAB) classification subtypes and correlate them with hematological parameters including red cell count (RBC) , hemoglobin, white cell count (WBC) , platelets count, blast cell percent and hematological response to the induction chemotherapy.
Patients and Methods:
EDTA anticoagulated peripheral blood (PB) sample of 2.5 ml and /or bone marrow (BM) aspirate samples of 0.5 ml from 25 cases of de novo AML including 14 male and 11 female with mean of age of 37.56±18.07 were included from June to November 2012.Hematological parameters including RBC, hemoglobin, WBC, platelets count, blast cell percent were obtained from the case file of the patients where they were done by automated device (Cell-DYN, RUBY list) and diagnosed by cytomorphology by Leishman stain and cytochemistry by Sudan Black B and PAS stain and AML cases were classified according to the FAB criteria. Aberrant lymphoid antigens, (CD7) and (CD19) expression was explored by CyFlow½ multiparametric flow cytometry at diagnosis. The patients were evaluated after three weeks of one cycle of chemotherapy for complete remission.
Results:
CD7, CD19 and co-expression of CD7 and CD 19 were expressed in 40%, 16% and 12% of AML patients respectively. Statistically significant associations were found between aberrant CD7 expressions and age, low WBC, and early FAB AML (M1, M2) subtypes. Complete remission was achieved in 19 out of 25 patients (76%) with standard chemotherapy whereas six patients did not achieved complete remission; three of them had aberrant CD7 expression (two of them died during induction therapy) and the other one had poor response to induction therapy.CD7 was detected in 7 patients; 6 of them were male. There is no statistically significant association between aberrant CD7 expression and hepatosplenomegaly and treatment response to one cycle of chemotherapy (P value > 0.05).Also there is no relation between CD7 expression and percent of blast cells.
Conclusions:
1. The incidence of aberrant expression of CD7, CD19 and both CD7, and CD19 were 40%, 16%, and12% respectively. 2. CD7 was detected mainly in males whereas CD19 was distributed among males and females. 3. Total WBC count and malignant cell percent were lower in patients harboring aberrant expression compare to those without aberrant expression. 4. CD7 was mainly detected in early FAB classification (M1 and M2). 5. Six out of twenty five AML patients had no response to standard therapeutic regimen; three of them were harboring CD7 and no one had CD19 alone, thus we may propose that CD7 was associated with poor response to induction therapy.
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Expression of CD10, CD13, CD19, CD20, CD22, CD33, CD34 and CD45 on B Lymphoblasts in Bone Marrow Aspirate of Adult Patients with Newly Diagnosed Acute Lymphoblasticleukemia Using Multicolor Flow Cytometry
Subh S Al-Mudallal
January-June 2014, 3(1):47-50
Background:
According to the 2008 WHO classification, the diagnosis of ALL relies on morphological demonstration of presence of 25% or more blast cells in bone marrow in ALL, and to consciously decide on the B lymphoid lineage of the blast cell population.
Objectives:
To evaluate the expression of CD10, CD13, CD19, CD20, CD22, CD33, CD34 and CD45 on B lymphoblasts in bone marrow aspirate of adult patients with newly diagnosed acute lymphoblastic leukemia using multicolor flow cytometry.
Materials and methods:
This is a cohort study which included 14 newly diagnosed adult patients with B ALL from April 2012 to May 2013. Flow cytometry analysis for CD10, CD13, CD19, CD20, CD22, CD33, CD34 and CD45 was carried out in a private laboratory on bone marrow aspirate samples using 2-laser, 4-color PARTEC Cube6 and using De Novo FCS Express version 4 Flow Cytometry software. The sensitivity of fluorescent detectors was monitored using standard beads according to the manufacturer's recommendations and normal lymphoid cells within specimens served as internal positive and negative controls. Samples with blasts that aberrantly expressed CD13 or CD33 were tested with SBB cytochemical stain.
Results:
The range and median percentages for B lymphoblasts, cells in the CD45dim versus low SSC gate, CD10, CD19, CD20, CD22, CD34, CD13 and CD33were68-99 and 85, 74-99 and 86, 41-88 and 68, 60-99 and 80, 48- 65 and 57, 48-99 and 75, 71-98 and 83, 20-51 and 37, and 20-43 and 30% respectively.
Conclusions:
FC testing should be accompanied by proper morphological evaluation. For diagnosis of B ALL, start with serial gating using FSC versus SSC, then CD45 versus SSC, then CD34 versus SSC, then using CD19, CD10 and CD22 to confirm the lineage of blasts as B lymphoid.
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Changes of immunological, Cytogenetic & Hematological Profiles in Chronic Myeloid Leukemia Treated with Imatinib Mesylate
Almotessembellah Abdalruhman
January-June 2014, 3(1):35-41
Background
: Chronic myeloid leukemia (CML) (also known as chronic myelogenous leukemia, chronic granulocytic leukemia) is a clonal disease that results from an acquired genetic change in a pluripotential hemopoietic stem cell .Imatinib mesylate, is an inhibitor of the family of ABL kinases, these proteins serves a complex role in cell cycling and is important in lymphopoiesis, and has been shown to have impressive clinical activity in chronic myeloid leukemia
Objectives
: to assess the immunological, cytogenetic, and hematological parameters in patients with chronic myeloid leukemia on imatinib therapy
Material and methods
: Thirty one CML patients treated with imatinib mesylate at standard dosage were enrolled in this prospective study. All patients were seen in the Department of hematology at the national center of hematology /Almustansiriya University for six months from February to July 2012.Data on blood cell counts and blood film, serum levels of IgG, IgA ,IgM ,C3 and C4;assessed by radial immundiffusion method,
Results
: Twenty four out of thirty one CML patients treated with the imatinib regimen (those had done FISH study) reached a complete cytogenetic response. Reduction percent were 19.35% for IgG, 16.13% for IgM and 9.68% for IgA, six patients had IgG inferior to normal laboratory range, five patients presented a reduction of IgM and three patients had IgA lower than normal range. No significant correlation of immunoglobulin levels compared to duration of treatment
Conclusion
: Imatinib mesylate can induce complete cytogenetic response in a high percent of CML patients. Cytogenetic response correlates well with duration of treatment .there is insignificant reduction of immunoglobulins level specially IgM. There is disturbance in the serum level of complement components (C3 &C4).There is insignificant reduction in the absolute count of lymphocytes in correlation with duration of treatment
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Incidence of Transfusion-Transmitted Viral Infections in the Hematology Department of Baghdad Teaching Hospital
Hassanain Hani Hassan, Ali M J Almodhaffar
January-June 2014, 3(1):14-22
Background:
Blood and blood product transfusion is vital for patients with hematological disorders including patients with chronic anemia and those with hematological malignancies but it carries the risk of transmission of infection. Hepatitis B, hepatitis C, and Human Immunodeficiency Virus (HIV) are the most prevalent of these transfusion-transmitted infections and should be screened for before transfusion for these patients.
Objectives:
The purpose of this study was to determine the rate of post-transfusion seroconversion of viral hepatitis B, C and HIV in the patients under the study.
Patients and methods:
A total of sixty-six consecutive patients (31 males and 35 females) were enrolled into this study between April 2011 and April 2012. These patients were admitted to the Hematology Department of Baghdad Teaching Hospital for management. These include twenty-six patients with acute myeloid leukemia (AML), sixteen patients with Non-Hodgkin's lymphoma (NHL), eleven patients with acute lymphoblastic leukemia, and two patients from each of the following hematological disorders: chronic lymphocytic leukemia (CLL), hairy cell leukemia, multiple myeloma, myelodysplastic syndrome, myelofibrosis and thrombotic thrombocytopenic purpura(TTP) with one patient with idiopathic vitamin K deficiency. Serology for HBsAg, anti-hepatitis C virus antibodies and HIV 1 & 2 was done for them before transfusion of any blood or blood product at diagnosis,3months and 6 months later to determine the rate of seroconversion of hepatitis B, C and HIV
Results:
Seroconversion for HBsAg was seen in nine patients with seroconversion rate of 13.6 %, while the serology for both hepatitis C and HIV were negative with sero-prevalence of (0%). The seroconversion for HBsAg was seen in 3 patients (9.4%) of lymphoid malignancies ( 1 with CLL , and 2 with NHL ) , 5 patients (16.1%) of myeloid malignancies (all of them had AML ) , and one patient with TTP . There is no statistically significant association between age , gender , type of hematological malignancies (lymphoid vs myeloid) and the type of the blood product transfused with or without HBsAg seroconversion (P value > 0.05). A statistically significant association was seen between clinical jaundice and seroconversion of HBsAg (which was seen in 7 out of nine patients who have seroconversion (77.8%) (and 2 of the patients who have no seroconversion (3.5%)) and biochemically between the mean of SGOT , SGPT , alkaline phosphatase and total serum bilirubin and seroconversion of HBsAg.
Conclusion:
This study showed that transfusion-transmitted hepatitis B seroconversion was more prevalent in our patient than hepatitis C and HIV which mandates more stringent preventive measures including appropriate donor selection , improved diagnostic testing methods that detect the virus at early period such as Nuclear Amplification testing (NAT) testing , use of pathogen reduction and inactivation methods to ensure blood safety and use of hepatitis B vaccination especially in immunocompromised patients with hematological malignancies
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Case Report: Myeloid Sarcoma : A Report of Two Cases
Bassam Francis Matti, Hassanain Hani Hassan
January-June 2014, 3(1):66-72
A 33 years old male was diagnosed as acute myeloid leukaemia on top of MPD at July ,2011.He received '3- 7' regimen with CR followed by 2 cycles of HiDAC chemotherapy and maintained on Gleevecô daily and stopped at January , 2012 because of frank relapse .He admitted in April ,2012 with disturbed level of consciousness Lab. Investigations showed:
CBP at 26-4-2012
PCV 0.16 WBCs 272 × 109 with 98% blasts Basophilia Platelets 16 × 109 FISH Study for BCR-ABL t(9,22)is detected in 99% of cells
Imaging : CT Brain
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H.Pylori Associated with Immune Thrombocytopenic Purpura
Baan A Mtashar, Zeyad A Shabeeb, Zainab F Ashoor
January-June 2014, 3(1):42-46
Background:
Idiopathic thrombocytopenic purpura (ITP) is a hematological disorder characterized by sensitization of platelets by autoantibodies leading to platelet destruction.). Recently, H. pylori has been found to be associated with ITP and its eradication has shown improvement in platelet count
Objective:
To determine the association of
Helicobacter pylori
infection in patients presenting with idiopathic thrombocytopenic purpura (ITP).
Materials and Methods:
From October 2012 to March 2013, fifty adult patients with ITP and fifty age and sex matched healthy controls were investigated for the presence of
H. pylori
infection by
Helicobacter pylori
detection IgG and IgM antibody using ELISA method.
Results:
H. pylori
IgM and
H. pylori
IgG: twelve patient (22%) and thirty nine patients (78%), of ITP patients had a positive expression
H. pylori
IgM and
H. pylori
IgG, respectively compared with three (6%) and thirty one (62%) patient in the control group
.
There was statistically significant difference between the mean of ITP patients and healthy control groups.
Conclusion:
The study confirms the existence of an association between
H. pylori
infection and ITP.
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Case Report: Bone Marrow Involvement as Initial Diagnosis of Metastatic Breast Cancer
Alaadin Sahham Naji, Zakka Noory
January-June 2014, 3(1):62-65
Bone marrow examination is commonly used in the evaluation of hemato-oncological disorders and in patients with cancer of solid organs to detect metastases. We present a 68 years old female with anemia, high ESR, minimal axial bone pain, weight loss. These features mimic and raise suspicion of myeloma, however in this case diagnosis was done retrogradely from the bone marrow, despite her aspirate does not show much of that typical cluster of non hemopoietic, her bone marrow biopsy showed extensive fibrosis with cluster of non hemopoietic elements. So bone marrow aspiration and trephine biopsy are an effective and cheap method for evaluating metastatic bone marrow tumors.
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Assessment of cytogenetic response after treatment with imatinib mesylate in patients with chronic phase chronic myeloid leukemia
Alaa Fadhil Alwan
January-June 2014, 3(1):56-61
Background
: Chronic myeloid leukemia (CML) is a myeloproliferative disorder affecting hematopoietic stem cells and affects predominantly granulocyte progenitor line. It is characterized by acquired chromosomal abnormality which called the Philadelphia chromosome (Ph+) in 95% of cases. Imatinib mesylate is a powerful and selective competitive inhibitor of BCR-ABL tyrosine kinase. Imatinib mesylate is the first molecular target therapy for the treatment of CML.
Objectives:
The aim of this study was to evaluate the cytogenetic response in 58 patients with CML in chronic phase treated with imatinib mesylate.
Materials and methods
: A prospective study conducted at the national center of hematology /Almustansiriya University, Baghdad, Iraq from April 2011 to December 2013, fifty-eight patients with CML in chronic phase (32 male and 26 female) were enrolled in this study. All patients were carrying the BCR-ABL fusion gene and treated with Imatinib mesylate (Glivicô Novartis) at 400 mg daily for at least 12 months
Results:
There were 32 male and 26 female with male to female ratio (1.1:1).the median age was 36 year (range 14-64 years). The median duration of treatment with imatinib was 18 months (range 12 to 32 months). Complete hematologic responses (CHR) were attained in 54 of 58 (93%) patients treated during the first 3 months of imatinib therapy, where 11 (19%) of patients reached CHR after 1 month, 35 (60%) got CHR within 2 months of treatment with imatinib. Cytogenetic response rates to imatinib therapy at 6 and 12months. : Major cytogenetic response achieved in 35 (70%), 48 (92.3%) patients and minor cytogenetic response attained in 15 (30%), 4 (7.7%) patient at 6, 12 month respectively. Imatinib was usually safe and well tolerated. The vast majority of adverse effects were grade 1 and 2 and bone pain was the most common (86.2%).
Conclusion
: After a median follow-up of 18 months, this study confirm that imatinib therapy induced induce durable and sustained hematological and cytogenetic responses in a high proportion of patients with chronic-phase CML. the response rates of imatinib therapy were similar to those reported in other countries. Imatinib was safe and well-tolerated with manageable side effects.
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Assessment of Soluble Fas in Patients with Chronic Myeloid Leukemia
Sahar Radhi Yasir, Maysoon Ali Saleem, Bassam Francis Matti
January-June 2014, 3(1):51-55
Background:
Soluble Fas was produced from the cell surface of malignant cells in a form lacking the 21 amino acid residues containing the transmembrane domain by alternative splicing. Trimerization of Fas receptor can be inhibited by soluble Fas (sFas) that act as decoys, binding FasL and preventing association with transmembrane Fas. So it is supposed to act as a FasL inhibitor to bind Fas and prevent Fas-mediated apoptosis.
Objective:
to evaluate serum soluble Fas (sFas) level in patients with chronic myeloid leukemia
Materials and Method:
Serum levels of sFas were measured by ELISA method after venous blood was collected from 56 CML patients (newly diagnosed and optimally treated) and 28 healthy subjects as control group. Absorbance was read at a wave length of 450nm using ELISA reader. Soluble Fas level was then calculated by plotting the optical density (O.D.) of each sample against the concentration in the standard curve.
Results:
There were no significant increases in serum sFas patient compared to healthy control with P=0.09. When the mean sFas concentration was obviously highest in newly diagnosed (1163.6pg/ml) followed by optimally treated (1021.7 pg/ml) and lowest in healthy control (970.1pg/ml).
Conclusion:
Production of sFas in tumor patients may be a key mechanism to inhibit Fas-mediated apoptosis. The identification of sFas levels as a predictor of outcome in malignant disease further establishes a connection between Fas loss-of-function and tumor progression.
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Estimation of Interleukin-10 Levels as a Predictive Factor in Iraqi Non-Hodgkin's Lymphoma Patients
Hassnien S AL-Hashemi, Zeyad A Shabib, Majed M Mahammod
January-June 2014, 3(1):23-30
PMID
:0
Background:
Interleukin-10 (IL-10) is a pleiotropic cytokine produced by type 2 helper cells (TH2), monocytes , macrophages, and neoplastic B lymphocytes IL-10 production has a strong immunosuppressive effect, and it acts as an effective stimulator for B-cells .IL-10 level increase in Non-Hodgkin lymphoma (NHL) patients ;moreover , this was associated with poor prognosis .
Objectives:
to assess the level of interleukin-10 in patients with non-Hodgkin lymphoma and to correlate its level with prognosis.
Materials and Methods:
The present study was carried out to evaluate IL-10 level in the sera of 46 NHL Iraqi patients, in addition to a control group involving 46 matched apparently healthy subjects using serological method the enzyme-linked immunosorbent assay (ELISA).This study was carried out at the National center of hematology from January to June 2013.
Results:
there were 31 (67%) patients with age group ≥50 years and 15 (33%) patients with age group <50 years. The mean age of NHL patients was 52.74 years ranging from 17 to 80 years? Moreover, the study noticed a male predominance and male to female ratio was 1.7:1.When the patients of NHL compared with control, the levels of interleukin 10 were markedly increased in patients with NHL (18.67 ± 4.12pg/ml vs. 3.4 ± 0.73pg/ml) (p<0.05). Higher levels of IL-10 were noticed in the advanced stage patients of compared to early stages (33.1 ± 4.5 pg. /ml vs. 13.4 ± 2.7 pg. /ml) (P<0.05).
Conclusion:
In the highest age of NHL patients was noticed and it might indicate the increasing rates of NHL patients among older population. in addition increasing level of IL10 is associated with increasing age range and also correlated positively with aggressive NHL cases that might indicates the predictive pivotal role of this cytokine.
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Evaluation of Leukemic Patient`s Compliance with Oral Chemotherapy in Baghdad City
Mohammed Salim, Mohammed Al-Jubouri, Ahmed Alsaffar
January-June 2014, 3(1):31-34
Background:
Noncompliance is a major obstacle to the effective delivery of health care. Estimates from the World Health Organization (WHO) indicate that only about 50% of patients with chronic diseases living in developed countries follow treatment recommendations
Objectives:
To evaluate the leukemic patient`s compliance with oral chemotherapy in Baghdad city.
Material and methods:
A descriptive study was carried out at Baghdad hospitals (Baghdad teaching hospital, and nursing home hospital). Started from 11 of December 2012 to 27 of June 2013. A non-probability (purposive) sample of 60 patients with leukemia and they were on oral chemotherapy. The questionnaire was designed and constructed by the researcher according to review of literatures and related studies. The content validity of the instrument was established through penal of (11) experts. Reliability of the patient compliance was determined by test-retest method which was estimated as average (r=0.851).Data was gathered by interview technique using the questionnaire format and data was analyzed by application of descriptive and inferential statistical methods.
Results
: Regarding age group (20-29) years was the larger group (31.7%). Nearly equal percent of male to female (48.3% and 51.7% respectively). Larger group (38.3%) of sample was primary school in educational level and (65%) of sample has less than one year illness duration. The result indicates the patient compliance with oral chemotherapy in which (55%) of patients was compliance and (45%) was not compliance with oral chemotherapy. The result also shows that there are no significant relationship at P>0.05 between (age groups, gender and educational level), and patient compliance to chemotherapy, which there are significant correlation at P<0.05 between patients duration of illness and patient`s compliance to chemotherapy.
Conclusion:
The results reveals that the majorities of study group were (20-29) years old, nearly equal percent of male to female, and primary school in educational level. The result indicated that (45%) of patients with leukemia are not compliance to oral chemotherapy.
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Haematology Clinico-pathological Exercise
January-June 2014, 3(1):73-74
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