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Fulltext Thalassaemia in Malaysia: a strategy for prevention

Ainoon O, Cheong SK

Malays J Pathol, 1994 Jun;16(1):23-7.
PMID: 16329572

In Malaysia, alpha-thalassaemia, beta-thalassaemia, haemoglobin (Hb) E, deltabeta-thalassaemia and Hb Constant Spring are prevalent. It has been estimated that 1 in 4 persons carries one of the above genetic abnormalities. In clinical practice, the major problems are: Hb Bart's hydrops fetalis (homozygous alpha(o)thalassaemia), homozygous 3(o)-thalassaemia, E-alpha thalassaemia and HbH disease. The laboratory procedures for diagnosis are standardised and the molecular basis of most of these genetic abnormalities are characterised. Thus it is possible to formulate a strategy for the detection and prevention of these disorders. The steps include the setting-up of population screening and genetic counselling service for the affected individuals, Society of Thalassaemias for public education and group support, and prenatal diagnosis with selective abortion of affected pregnancies. We embarked on such a programme between 1988 and 1992 in Kuala Lumpur General Hospital and hope to kindle similar effort in other state hospitals.
Fulltext Subtractive hybridization identifies stem cell-associated genes in an acute myeloid leukemia with poor prognosis

Ngiow Shin Foong, Maha Abdullah, Jasmine Lim, Cheong Soon-Keng, Seow Heng-Fong

Malaysian Journal of Medicine and Health Sciences, 2016;12(1):19-31.
MyJurnal

Introduction: Current prognostic markers have improved survival prediction, however, it has not
advanced treatment strategies. Gene expression profiling may identify biological markers suitable as
therapeutic targets. Leukaemia stem cell is associated with adverse outcome, however, its biological
characteristics are still being investigated. We observed higher in vitro cell viability in acute myeloid
leukaemia (AML) samples with poor prognosis, which may be stem cell related. Objective: The
objective of this study was to profile highly expressed genes in an AML sample of poor prognosis/high
viability and compare with a sample of good prognosis/low viability. Method: Subtractive hybridization
was performed on two AML samples with high blast counts (>80%), a poor prognosis, PP (disease free
survival, DFS12 months) sample. The PP sample had
higher CD34+ counts (73% vs 46%) and higher cell viability than the GP sample. cDNA libraries were
subsequently cloned and sequenced. Results: cDNA subtracted from the PP samples was identified
as genes active during fetal/embryonic development (LCOR, CNOT1, ORMDL1), HOX- related genes
(HOXA3, PBX3, SF3B1), hematopoiesis (SELL, IL-3RA) and aerobic glycolysis/hypoxia (PGK1,
HIGD1A) -associated genes. Majority of GP clones isolated contained genes involved in oxidative
phosphorylation, OXPHOS (COXs, ATPs, MTND4 and MTRNR2), protein synthesis (including
ribosomal proteins, initiating and elongation factors), chromatin remodeling (H2AFZ, PTMA), cell
motility (MALAT1, CALM2, TMSB4X), and mitochondria (HSPA9, MPO) genes. Conclusion: Thus,
the PP sample exhibited stem cell-like features while the GP sample showed cells at a high level of cell
activity. These genes are potential prognostic markers and targets for therapy.
Fulltext Differential expression patterns of leukaemia associated genes in leukaemia cell lines compared to healthy controls

Ang Pei-Shen, Rajesh Ramasamy, Noor Hamidah Hussin, Cheong Soon-Keng, Seow Heng-Fong, Maha Abdullah

Malaysian Journal of Medicine and Health Sciences, 2016;12(1):0-0.
MyJurnal

Introduction: The phenotype and genotype of cancer cells portray hallmarks of cancer which may
have clinical value. Cancer cell lines are ideal models to study and confirm these characteristics. We
previously established two subtracted cDNA libraries with differentially expressed genes from an
acute myeloid leukaemia patient with poor prognosis (PP) and good prognosis (GP). Objective: To
compare gene expression of the leukaemia associated genes with selected biological characteristics
in leukaemia cell lines and normal controls. Methodology: Expression of 28 PP genes associated
with early fetal/embryonic development, HOX-related genes, hematopoiesis and aerobic glycolysis/
hypoxia genes and 36 GP genes involved in oxidative phosphorylation, protein synthesis, chromatin
remodelling and cell motility were examined in B-lymphoid (BV173, Reh and RS4;11) and myeloid
(HL-60, K562) leukaemia cell lines after 72h in culture as well as peripheral blood mononuclear cells
from healthy controls (N=5) using semi-quantitative polymerase chain reaction (PCR) method. Cell
cycle profiles were analysed on flow cytometry while MTT cytotoxicity assay was used to determine
drug resistance to epirubicin. Results: Genes expressed significantly higher in B-lymphoid leukaemia
cell lines compared to healthy controls were mostly of the GP library i.e. oxidative phosphorylation
(3/10), protein synthesis (4/11), chromatin remodelling (3/3) and actin cytoskeleton genes (1/5). Only
two genes with significant difference were from the PP library. Cancer associated genes, HSPA9 and
PSPH (GP library) and BCAP31 (PP library) were significantly higher in the B-lymphoid leukemia cell
lines. No significant difference was observed between myeloid cell lines and healthy controls. This
may also be due heterogeneity of cell lines studied. PBMC from healthy controls were not in cell cycle.
G2/M profiles and growth curves showed B-lymphoid cells just reaching plateau after 72 hour culture
while myeloid cells were declining. IC50 values from cytotoxicity assay revealed myeloid cell lines had
an average 13-fold higher drug resistance to epirubicin compared to B-lymphoid cell lines. Only CCL1,
was expressed at least two-fold higher in myeloid compared to B-lymphoid cell lines. In contrast,
MTRNR2, EEF1A1, PTMA, HLA-DR, C6orf115, PBX3, ENPP4, SELL, and IL3Ra were expressed
more than 2-fold higher in B-lymphoid compared to myeloid cell lines studied here. Conclusion: Thus,
B-lymphoid leukaemia cell lines here exhibited active, proliferating characteristics closer to GP genes.
Higher expression of several genes in B-lymphoid compared to myeloid leukaemia cell lines may be
useful markers to study biological differences including drug resistance between lineages.
Fulltext Induced pluripotent stem cells in research and therapy

Teoh HK, Cheong SK

Malays J Pathol, 2012 Jun;34(1):1-13.
PMID: 22870592 MyJurnal

Induced pluripotent stem cells (iPSC) are derived from human somatic cells through ectopic expression of transcription factors. This landmark discovery has been considered as a major development towards patient-specific iPSC for various biomedical applications. Unlimited self renewal capacity, pluripotency and ease of accessibility to donor tissues contribute to the versatility of iPSC. The therapeutic potential of iPSC in regenerative medicine, cell-based therapy, disease modelling and drug discovery is indeed very promising. Continuous progress in iPSC technology provides clearer understanding of disease pathogenesis and ultimately new optimism in developing treatment or cure for human diseases.
Fulltext Leukaemic stem cells: drug resistance, metastasis and therapeutic implications

Wong RS, Cheong SK

Malays J Pathol, 2012 Dec;34(2):77-88.
PMID: 23424769 MyJurnal

Although there have been many new developments in the treatment of leukaemia with the use of new anti-leukaemic agents and stem cell transplantation, drug resistance and treatment failure remain a great challenge for the attending physician. Several studies have suggested that leukaemic stem cells (LSCs) play a pivotal role in chemoresistance and metastasis and the mechanisms by which these cells do so have also been elucidated. There is increasing evidence to show that there exists a large pool of therapeutic targets in LSCs and that the eradication of these cells is feasible with some promising results. This article gives an overview of different types of cancer stem cells (CSCs) derived from various types of leukaemia, the mechanisms by which LSCs contribute to drug resistance and metastasis and some recent advances in targeted therapy against LSCs.
Fulltext Ocular relapse in a patient with acute lymphoblastic leukemia: a case report

Cheong SK, Tan PL

Med J Malaysia, 1985 Jun;40(2):132-5.
PMID: 3869273

A female Malay patient with acute lymphoblastic leukemia in remission for one year developed bilateral visual loss. The ophthalmoscopic appearance showed infiltration of optic nerves and retinae by leukemic cells. At that time, her blood, bone marrow and cerebral spinal fluid remained normal. She had received prophylactic cranial irradiation and intrathecal methotrexate as part of the treatment programme. It was likely therefore she had a leukemic relapse from a pharmacologic sanctuary in the eyes. She responded to local irradiation but did not regain her sight. Three months later, she had a bone marrow relapse.
Fulltext Protein and gene expression of leukemia stem cells markers in acute myeloid leukemia

Amrina Mohamad Amin, Maha Abdullah, Sabariah Md Noor, Raudhawati Osman, Wan Hayati Mohd Yaacob, Cheong Soon Keng

Malaysian Journal of Medicine and Health Sciences, 2019;15(108):23-23.
MyJurnal

Introduction: Acute myeloid leukaemia (AML) is a clonal haematological neoplasm characterised by proliferation of immature myeloid cells in the bone marrow resulting in impairment normal cell development in bone marrow. This leads to anaemia, thrombocytopenia and neutropenia. AML primarily affects older adults, with a median age at diagnosis of 69 years but is also seen in all other age groups. AML is recognized as a kind of cancer with marked heterogeneity in both biology of the cells and reactions to treatment. Treatment with intensive chemotherapy regi-mens of adult AML patients who are ≤ 60 years old results in hematologic remission in about 35% of patients, but at least 30% of these patients will experience a relapse. Mechanism leading to early relapse is still unclear. Leukaemia stem cell (LSC) is shown to correlate with poor prognosis. Biomarkers such as aldehyde dehydrogenase (ALDH) and CD34+CD38- have been identified as potential LSC biomarkers in previous studies. The objective of this study is to examine the expression of such markers for LSC and determine the association. Methods: Peripheral blood or bone marrow samples from untreated, newly diagnosed acute myeloid leukemias of all age, gender and race were collect-ed from Hospital Melaka and Kelang. Diagnosis of AML is based on WHO classification which include morphology, cytochemistry, immunophenotyping and cytogenetics. Mononuclear cells were isolated from bone marrow aspirate samples by gradient density centrifugation on Ficoll-Hypaque. Immunophenotyping using CD13, CD14, CD33, CD34, CD38 and ALDH were carried out to identify the presence and proportion of the various populations of inter-est. Results: There was a strong, positive correlation between ALDH and CD34+CD38- cell population, which was statistically significant (rs = 0.5989, p< 0.05). Conclusion: The strong correlation of ALDH activity and CD34+CD38- expression supported the potential of these biomarkers to identify LSCs cell in AML patients. However, due to the heterogeneity of AML, further studies using more markers and larger sample size are needed to determine the validity and to correlate with disease-free survival rate of AML patients.
Role of mesenchymal stem cells in leukaemia: Dr. Jekyll or Mr. Hyde?

Wong RS, Cheong SK

Clin Exp Med, 2014 Aug;14(3):235-48.
PMID: 23794030 DOI: 10.1007/s10238-013-0247-4

Mesenchymal stem cells (MSCs) have captured the attention of researchers today due to their multipotent differentiation capacity. Also, they have been successfully applied clinically, in the treatment of various diseases of the heart and musculoskeletal systems, with encouraging results. Their supportive role in haematopoiesis and their anti-inflammatory and immunomodulatory properties have enhanced their contribution towards the improvement of engraftment and the treatment of graft-versus-host disease in patients receiving haematopoietic stem cell transplantation. However, there is a growing body of research that supports the involvement of MSCs in leukaemogenesis with several genetic and functional abnormalities having been detected in the MSCs of leukaemia patients. MSCs also exert leukaemia-enhancing effects and induce chemotherapy resistance in leukaemia cells. This paper addresses the key issues in the therapeutic value as well as the harmful effects of the MSCs in leukaemia with a sharp focus on the recent updates in the published literature.
Using digital photography to facilitate learning for medical students

Loh KY, Boo NY, Cheong SK

Med Educ, 2012 Nov;46(11):1120-1.
PMID: 23078710 DOI: 10.1111/medu.12043
Fulltext Observation of dendritic cell morphology under light, phase-contrast or confocal laser scanning microscopy

Tan YF, Leong CF, Cheong SK

Malays J Pathol, 2010 Dec;32(2):97-102.
PMID: 21329180 MyJurnal

Dendritic cells (DCs) are professional antigen presenting cells of the immune system. They can be generated in vitro from peripheral blood monocytes supplemented with GM-CSF, IL-4 and TNF alpha. During induction, DCs will increase in size and acquire multiple cytoplasmic projections when compared to their precursor cells such as monocytes or haematopoietic stem cells which are usually round or spherical. Morphology of DCs can be visualized by conventional light microscopy after staining or phase-contrast inverted microscopy or confocal laser scanning microscopy. In this report, we described the morphological appearances of DCs captured using the above-mentioned techniques. We found that confocal laser scanning microscopy yielded DCs images with greater details but the operating cost for such a technique is high. On the other hand, the images obtained through light microscopy after appropriate staining or phase contrast microscopy were acceptable for identification purpose. Besides, these equipments are readily available in most laboratories and the cost of operation is affordable. Nevertheless, morphological identification is just one of the methods to characterise DCs. Other methods such as phenotypic expression markers and mixed leukocyte reactions are additional tools used in the characterisation of DCs.
Autoimmune thrombocytopenia and neutropenia after autologous peripheral blood stem cell transplantation

Wahid FS, Cheong SK, Sivagengei K

Acta Haematol., 2002;107(4):237-8.
PMID: 12053154
Circulating tissue factor, tissue factor pathway inhibitor and D-dimer in umbilical cord blood of normal term neonates and adult plasma

Tay SP, Cheong SK, Boo NY

Blood Coagul Fibrinolysis, 2003 Feb;14(2):125-9.
PMID: 12632021

The investigation of many hemostatic defects in newborns is restricted by the lack of normal reference values. The coagulation system of the neonate differs in many ways from that of the adult. The present study was designed to compare the concentrations of tissue factor (TF), tissue factor pathway inhibitor (TFPI) and D-dimer (DD) in the umbilical cord blood of healthy newborns and in adult plasma. TF antigen was quantified using an in-house enzyme-linked immunosorbent assay, whereas TFPI and DD levels were measured with commercial kits. The mean TF level in cord blood (mean standard deviation, 183.94 103.63 pg/ml) was significantly higher ( = 0.008) than that in adults (136.64 65.09 pg/ml). Cord blood exhibited enhanced fibrinolysis, as was reflected by a significantly higher level of DD (924.57 733.87 ng/ml, 0.001) than that in adults (45.57 17.21 ng/ml). Conversely, cord blood (30.88 10.16 ng/ml) demonstrated significantly lower ( 0.001) TFPI levels than that in adults (55.77 21.16 ng/ml). However, no significant differences of these three hemostatic markers were noted between both gender groups in newborns and adults. Our findings indicate that an active and dynamic state of hemostasis exists in cord blood, as the fluidity of cord blood remains preserved in the presence of birth injury.
In vitro differentiation of mesenchymal stem cells into mesangial cells when co-cultured with injured mesangial cells

Wong CY, Tan EL, Cheong SK

Cell Biol Int, 2014 Apr;38(4):497-501.
PMID: 24375917 DOI: 10.1002/cbin.10231

Mesangial cells are one of the three major cell types of the kidney glomerulus that provide physical support for the glomerular capillary lumen of the kidney. Loss of mesangial cells due to pathologic conditions, such as glomerulonephritis and diabetic nephropathy, can impair renal function. Mesenchymal stem cells (MSC) are attractive candidates for kidney repair therapy since they can enhance recovery and protect against kidney failure. MSC can differentiate into mesangial cells in vivo. We have investigated the ability of MSC to differentiate into mesangial cells in vitro; they were co-cultured with oxidant-injured mesangial cells before being analysed by flow cytometry and for contractility. MSC co-cultured with injured mesangial cells had a mesangial cell-like morphology and contracted in response to angiotensin II. They expressed CD54(-) CD62E(+) in direct contrast to the CD54(+) CD62E(-) of pure MSC. In conclusion, MSC can differentiate into mesangial cells in vitro when co-cultured with injured mesangial cells.
Study of EEG alpha wave response on the effects of video-guided deep breathing on pain rehabilitation

Ramalingam V, Cheong SK, Lee PF

Technol Health Care, 2023;31(1):37-46.
PMID: 35723127 DOI: 10.3233/THC-213531

BACKGROUND: Athletes with chronic ankle pain (CAP) are more inclined to suffer from physical and psychological pain depending on the severity of the injuries, which might trigger the powerless feeling on future sports participation. Therefore, an efficient and simple method is useful to integrate into conventional physiotherapy (CP) for maintaining mental wellness.
OBJECTIVE: This research aimed to verify the effects and progress of video-guided deep breathing (DB) integrated into CP through study on the changes of alpha waves and pain scale.
METHODS: Alpha waves were recorded using an electroencephalogram (EEG) and a visual analogue scale (VAS) to assess pain intensity before and after the intervention (6 weeks). Thirty CAP participants were recruited and randomly assigned to two groups: group A for video-guided DB integration into their CP and group B for CP. The effects of pre and post intervention were analyzed using a paired t-test with statistical significance set at p< 0.05.
RESULTS: Profound results from the research have shown that the participants who received both the DB+CP revealed a significant increase in alpha wave (p< 0.05) at occipital region.
CONCLUSION: The significant result reveals an increase in alpha waves in the occipital region after 6 weeks and indicates that the video-guided DB with a smartphone application is able to produce a change in CAP participants. This supports the DB integration to the CP for altering the pain perception.
Effect of six-week short-duration deep breathing on young adults with chronic ankle instability-a pilot randomized control trial

Ramalingam V, Cheong SK, Lee PF

BMC Sports Sci Med Rehabil, 2023 Nov 15;15(1):155.
PMID: 37968738 DOI: 10.1186/s13102-023-00758-5

BACKGROUND: Chronic ankle instability (CAI) is the most common injury in youth sports, which leads to psychological stress from doubting their performance. Cost effective and easy to access tool to reduce the stress among this target group are desired. Therefore, the purpose of this study was to investigate the effect of adding on intervention with short-duration deep breathing (SDDB) alongside with conventional physiotherapy (CP) among young adults with chronic ankle instability (CAI).
METHODS: Total of 30 CAI participants attended physiotherapy, who were randomly assigned into control and experimental groups. The participants in the experimental group received combined intervention (SDDB + CP), and the control group received CP for 6 weeks. The effectiveness of interventions was assessed at 3 intervals with a battery of questionnaires (Visual Analog Score, Cumberland Ankle Instability Tool, Mindful Attention Awareness Scale, and Oxford Happiness Questionnaire) at the end of week 3, week 6, and week 12 as follow-up. A two-way repeated measures of ANOVA was applied to report the statistical significance at p
Fulltext Comparing the ability of freshly generated and cryopreserved dendritic cell vaccines to inhibit growth of breast cancer in a mouse model

Radhakrishnan AK, Sim GC, Cheong SK

Biores Open Access, 2012 Oct;1(5):239-46.
PMID: 23515111 DOI: 10.1089/biores.2012.0229

Repetitive vaccinations with dendritic cell (DC)-based vaccines over long periods of time can break pre-existing tolerance to tumors and achieve clinically relevant immune response. This requires a large number of DCs to be generated under good manufacturing protocol, which is time- and cost intensive. Thus, producing a large numbers of DCs at one time point and cryopreserving these cells in ready-for-use aliquots for clinical application may overcome this constraint. This could also reduce batch-to-batch variations. In this study, we generated DCs from bone marrow obtained from BALB/c mice. Some of the generated DCs were cryopreserved before conducting various tests. There were no significant differences in the morphology and phenotype between cryopreserved and freshly generated DCs. Both types of DCs pulsed with tumor lysate (TL) from 4T1 murine mammary cancer cells (DC+TL) possessed a similar capacity to stimulate the proliferation of T-cells. In addition, cryopreserved and fresh DC pulsed with TL showed similar tumor growth inhibition patterns. Both DCs induced initial retardation of tumor growth (p<0.05) and prolonged the survival (p<0.05) of tumor-bearing mice treated with DC+TL as compared with nontreated control mice. Cryopreserved DCs shared similar therapeutic efficacy to fresh DCs, and this finding lends supports the routine use of cryopreserved DCs in future clinical trials.
Fulltext Molecular responses during chemotherapy in acute myeloid leukemias in predicting poor-response to standard chemotherapy

Maha A, Cheong SK, Leong CF, Seow HF

Malays J Pathol, 2009 Dec;31(2):81-91.
PMID: 20514850 MyJurnal

Signal transduction pathways are constitutively expressed in leukaemic cells resulting in aberrant survival of the cells. It is postulated that in cells of chemo-sensitive patients, chemotherapy induces apoptotic signals leading to cell death while survival signals are maintained in cells of chemo-resistant patients. There is very little information currently, on the expression of these mediators in patients immediately after chemotherapy initiation. We examined the expression pattern of proinflammatory cytokines, signaling molecules of the PI3K and MAPK pathways molecules and death receptor, DR5 on paired samples at diagnosis and during chemotherapy in acute myeloid leukaemia patients treated with cytosine arabinoside and daunorubicin. The results were correlated with remission status one month after chemotherapy. We found that in chemo-sensitive patients, chemotherapy significantly increased the percentage of cases expressing TNF-alpha (p = 0.025, n = 9) and IL-6 (p = 0.002, n = 11) compared to chemo-resistant cases. We also observed an increased percentage of chemo-sensitive cases expressing DR5 and phosphorylated p38, and Jnk. Thus, expression of TNF-alpha, IL-6, DR5, phospho-p38 and phospho-Jnk may regulate cell death in chemo-sensitive cases. In contrast, a significantly higher percentage of chemo-resistant cases expressed phospho-Bad (p = 0.027, n = 9). IL-beta and IL-18 were also found to be higher in chemo-resistant cases at diagnosis and during chemotherapy. Thus, expression of various cellular molecules in leukaemic blasts during chemotherapy may be useful in predicting treatment outcome. These cellular molecules may also be potential targets for alternative therapy.
Fulltext Transfected human mesenchymal stem cells do not lose their surface markers and differentiation properties

Yap FL, Cheong SK, Ammu R, Leong CF

Malays J Pathol, 2009 Dec;31(2):113-20.
PMID: 20514854 MyJurnal

In this study, we evaluated the biological properties of human mesenchymal stem cells transfected (hMSC) with a plasmid vector expressing human cytokine interleukin-12 (IL-12). Surface markers were analysed by immunophenotyping using flow cytometry. Differentiation capability was evaluated towards adipogenesis and osteogenesis. We demonstrated that successfully transfected hMSC retained their surface immunophenotypes and differentiation potential into adipocytes and osteocytes. These results indicate that hMSC may be a suitable vehicle for gene transduction.
Fulltext Dendritic cell distribution in lymphomas

Hussin HN, Zulkifli FN, Phang KS, Cheong SK

Malays J Pathol, 2009 Dec;31(2):105-12.
PMID: 20514853 MyJurnal

Dendritic cells (DC) are specialized antigen presenting cells (APC) that have important roles in host defenses and in generating anti-tumour immune response. Altered frequency and maturation of DC have been reported in malignant tumours. We studied the distribution and maturation status of DC by immunohistochemistry, on the formalin-fixed, paraffin-embedded lymph node tissues of 32 histologically diagnosed lymphomas and 40 inflammatory conditions that were retrieved from the Department of Pathology, UKM Medical Centre, Kuala Lumpur. Our study showed a significant reduction in the total DC counts in the lymphoma tissues compared to the inflammatory conditions. The mature and immature DC counts were both significantly reduced (p = 0.008 and 0.001 respectively), although a greater reduction was observed in mature DC numbers. We also observed compartmentalization of DC where the immature DC were seen within the tumour tissues and the mature DC were more in peri-tumoural areas. Our findings were similar to other reports, suggesting that reduced numbers of DC appears to be a factor contributing to lack of tumour surveillance in these cases.
Cell viability of acute myeloid leukaemia blasts in culture correlates with treatment outcome

Maha A, Cheong SK, Leong CF, Seow HF

Hematology, 2008 Feb;13(1):13-20.
PMID: 18534060 DOI: 10.1179/102453308X315762

Despite the advances in understanding the pathophysiology of acute myeloid leukaemia (AML), the cure rate for acute myeloid leukaemia patients remains low. Cytogenetic abnormalities and age are the prognostic factors that guide treatment decisions. However, many AML patients still die. The biological factors that influence treatment outcome are largely unknown. Thus, the objective of our study was to use the in vitro viability test to correlate with treatment outcome. Acute myeloid leukaemia blasts demonstrated differing ability to survive in culture. Our examination of blast phenotype at various days in culture showed two possible growth directions. First, cells underwent maturation by increased expression of CD16 and down-regulated CD34 (a haemopoietic stem cell marker). These cells also appeared to have undergone apoptosis. Alternatively, cells continued to survive in culture and maintained high expression of CD34. An MTT assay was carried out to determine viability after three days of culture. Lower optical density values were obtained for samples that underwent apoptosis and higher values were obtained for samples that survived in culture. Apoptosis was measured by Annexin V/propidium iodide staining. A comparison between results of MTT assay and duration of disease free survival revealed that a higher viability in vitro correlated significantly with shorter survival duration in the patient (R -0.761, p=0.002, n=13). Thus, this study further supports the hypothesis that AML patients with poor survival may be related to having blasts with a biologically more immature or stem cell-like nature.

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