Recent advances in diagnosis of acute myeloid leukaemia : a review.

Acute Myeloid Leukaemia is one of the commonest blood cancer in Malaysia. Recent advancements in molecular genetics have led to better understanding of AML pathogenesis which is crucial for patient stratification and therapeutic options. The advent of next generation sequencing (NGS) is now the vogue to interrogate acute myeloid leukaemia (AML) genome at single base pair resolution. In this review, we will discuss current approaches in the diagnosis of AML and the role of NGS in elucidation of genomic aberrations. Although NGS is anticipated to impact the new era of AML diagnosis, there are challenges in understanding mutations found in an AML genome and how this technology will be implemented in a diagnostic setting. NGS will pave the way for perso nalized medicine approach, with the hope of better response to tailored treatment regimen

Antigen expression pattern of acute promyelocytic leukaemia cases in Malaysia

Introduction: Acute Promyelocytic Leukaemia (APL) is associated with devastating coagulopathy and life threatening condition which requires immediate medical attention. It is crucial to establish an expedited diagnosis as early therapeutic intervention has led to optimal patient management. In this study, we assessed the type and frequency of antigen expressions in APL and correlated these findings with genetic studies. Methods: Multiparametric immunophenotyping was performed on 30 samples and findings were correlated with karyotypes, FISH for t(15;17) translocation and RT-PCR for PML-RARα for detection of breakpoint cluster regions (bcr1,bcr2 and bcr3). Results: On SSC/CD45, APL cells displayed high to moderate SSC, with the expression of CD33 (100%), CD13 (96.8%), cMPO (71%) but lacked CD34 (3.2%) and HLA-DR (9.7%). Aberrant expression of CD4 was seen in 12.9% and CD56 in 6.5% of the cases. A significant association between cumulative aberrant antigen expression and bcr1 were observed bcr1 (X2(2) =6.833,p.05) and (X2(2)=4.599,p>.05) respectively. Conclusions: Flow cytometry is a rapid and effective tool in detecting APL. It is interesting to note that there is significant association between cumulative aberrant antigen expression and genotype analysis. Further validation is required to corroborate this relationship

Aggressive variant large granular lymphocytic leukaemia: a case report

Clonal disorders of LGL may either be CD3+ CD56- or CD3- CD56+ phenotype and these have been designated as T-cell leukaemia (T-LGL) or natural killer cell (NK)-LGL leukaemia respectively. Clonality is usually demonstrated by clonal rearrangement of T-cell receptor gene rearrangement or identified by flowcytometry analysis. Most patients with T-LGL will have an indolent course. In this report we described an aggressiveness of disease in a patient with clonal CD3+ LGL leukaemia whose cells also co-expressed CD56 diagnosed by flowcytometry. The patient responded well to interrupt all standard risk protocol however succumbed to her disease while waiting for upfront stem cell transplant. This case highlights on both the classical laboratory findings of rare entity of disease as well as a review of the literature pertaining particularly on its management

Qualitative flow cytometric analysis of Malaysian myelodysplastic syndromes (MDS) patients.

Myelodysplastic syndromes (MDS) are a group of haematological malignancies categorized by ineffective hematopoiesis that result in dysplasia. Although morphological diagnosis is a traditional and standard technique that is used for the diagnosis of MDS, the heterogeneous blood and bone marrow characteristics of MDS patients can potentially obscure the right diagnosis. Thus, we have utilized flow cytometric immunophenotyping as a supportive mechanism to obtain a more accurate and faster method for detection of abnormal markers in MDS. Flow cytometry was used for analyzing bone marrow samples from newly diagnosed MDS patients to investigate the abnormal antigen expression patterns in granulocytic, monocytic, erythroid, lymphoid lineages and myeloid precursors. The results were compared with those obtained from cases that had Idiopathic Thrombocytopenic Purpura (ITP) as a control. The most common abnormality found in the granulocytic lineage was the decrease of CD10. Low expressions of CD13 were the most frequent abnormality in the monocytic lineage. The erythroid lineage was found to have low expression of CD235A+/CD71+, reduce of CD71 and decreased CD235a. In conclusion, this method is useful for confirming cases in which it is difficult to make a diagnosis by morphology

Thrombosis at unusual sites in reproductive age group-a case series

Thrombosis is one of the causes of morbidity and mortality in women of reproductive age group. Thrombosis at unusual sites may pose diagnostic and management dilemma for health care personnel. Teamwork and good communication provide the best modalities for maximum benefits to patients. Here with, we presented case a series of thrombosis at unusual sites seen and managed in our clinic. A 35 year-old Malay lady presented with left hemiparesis while she was on oestrogen based combined contraception pills (C-OCP). Imaging studies showed extensive venous thrombosis with bilateral acute cortical infarct. Thrombophilia screening of antiphospholipid syndrome were negative. She was put on anticoagulant and stopped 2 years after the incident. A 40 year-old Malay lady presented with abdominal discomfort, lethargy and massive splenomegaly. Bone marrow and trephine examination revealed primary myelofibrosis with positive JAK2617F. Imaging study showed chronic portal vein thrombosis with portal vein hypertension, complicated by gastro-oesophageal varices. She was put on hydroxyurea and later started on ruxolitinib with banding done over her gastro-oesophageal varices. A 26 year-old Malay lady presented with serositis, mouth ulcer and anaemia symptoms. Laboratory studies were positive for systemic lupus erythematosus and negative for antiphospholipid study. Imaging study showed long segment thrombosis of right internal jugular vein with surrounding subcutaneous oedema. She is currently stable on anticoagulants and steroid. Teamwork and holistic approach is practiced in the investigation and management to provide maximum benefits for patients

Donor chimerism and bcr-abl gene status following non-myeloablative peripheral blood stem cell transplantation in chronic myeloid leukaemia patients in HUKM

Objective: This study was done to determine the relationship between donor chimerism and the presence of bcr-abl gene in chronic myeloid leukaemia (CML) patients post-transplantation. Methods: The study population consisted of all CML patients who had undergone non-myeloablative peripheral blood stem cell transplant in Hospital Universiti Kebangsaan Malaysia (HUKM) during the study period. All patients had their bone marrow aspiration done at diagnosis and day 30, 60, 100, 130, 160 and 190 post-transplantation. The samples were analysed for bcr-abl transcript as well as chimerism status. Results: A total of nine cases underwent non-myeloablative peripheral blood stem cell transplant. All patients were transplanted during the chronic phase. One patient was found to show mixed chimerism at day 30 post-transplant coinciding with bcr-abl transcript disappearance. Six patients showed that full donor chimerism correlated with bcr-abl transcript disappearance. In one patient, chronic myeloid leukaemia transformed into acute myeloid leukaemia. Another patient had a graft failure. Conclusion: This observational cohort study showed that full chimerism is required for disappearance of bcr-abl transcript but one case showed disappearance of bcr-abl transcript at day 30 while full chimerism was not achieved

Clinical utility of extended monocytes parameters as a screening tool in suspected dengue infection

Introduction: Increased monocyte percentage and monocyte anisocytosis were suggested as new markers for dengue fever detection. This study aims to investigate and evaluate monocyte volume standard deviation (MoV-SD) and monocyte percentage (Mono %) parameters using Coulter automated haematology analyser as screening parameters in discriminating between dengue infection and other febrile illness. Methods: A cross-sectional laboratory analysis using suspected dengue fever patients were included in this study. The study was conducted in the Department of Pathology, Hospital Tuanku Jaafar Seremban from June 2016 until June 2017. Patients were classified into dengue positive and dengue negative based on dengue IgM and NS1 result. The diagnostic performance of MoV-SD and Mono % was analysed by receiver operating characteristic (ROC) curve analysis. The cut-off value of the MoV-SD and Mono % was determined and evaluated with the validation group. Chi-square test was used to assess the association between the parameters. Results: 88 (48.4%) from 182 samples were confirmed to have dengue infection. ROC curve analysis showed Mono % at cut off value of 10.5 % with area under the curve (AUC) of 0.869 with 84.1% sensitivity and 84% specificity (95% CI: 0.812-0.925) and MoV-SD cut off value at 22.2 (AUC 0.776, 80.7% sensitivity, 61.7% specificity, 95% CI: 0.709-0.843) are an excellent parameters in separating dengue positive and dengue-negative patients. A cut-off value of 10.5 of Mono % and 22.2 of MoV-SD were applied to the validation group showed 83.1%, 66.4% sensitivity and 84.9%, 77.3% specificity respectively. Conclusion: MoV-SD and Mono % parameters are a potential parameter for the screening of dengue infection in acute febrile illness patients with good specificity and sensitivity

Evaluation of anticoagulant property of aqueous and Ethanolic Extracts of Morinda citrifolia

Anticoagulant is an agent used to treat patients with cardiovascular diseases by preventing new clots formation. It works by inhibiting blood from clotting, therefore preventing progression of thrombosis. Heparin, the animal based polysaccharides is a widely used anticoagulant has many adverse effects. This study aimed to evaluate the effects of Morinda citrifolia Aqueous Extract (MCAE) and Ethanolic Extract (MCEE) on plasma coagulation in vitro. Platelet Poor Plasma (PPP) from fifty healthy volunteers was incubated with different concentrations of extracts (10, 20, 30, 40 and 50 mg mL-1) and subjected for clotting assays of Prothrombin Time (PT) and Activated Partial Thrombosplastin Time (APTT). The samples that been incubated by MCAE and MCEE showed prolongation of PT and APTT. These findings indicated that Morinda citrifolia extracts has anticoagulant effect in vitro and also suggest that it may become a potential plant based anticoagulant which is should be effective and safe for clinical need in dealing with patient with cardiovascular disorders