Concavity Point and Skeleton Analysis Algorithm for Detection and Quantization in Heavily Clumped Red Blood Cells

In practice, most hospitals use light microscope to examine the smeared blood for blood quantification. This visual quantification is subjective, laborious and time-consuming. Although automating the process is a good solution, the available techniques are unable to count or ignore the clumpy red blood cells (RBC). Moreover, clumping cell can affect the whole counting process of RBC as well as their accuracy. This paper proposes a new quantization process called concavity point and skeleton analysis (CP-SA) for heavily clump RBC. The proposed methodology is based on induction approach, enhanced lime blood cell by using gamma correction to get the appropriate edges. Then, splitting the clump and single cells by calculating each object area in pixel. Later, the quantification of clumpy cells with the proposed CP-SA method is done. This algorithm has been tested on 556 clump RBC taken from thin blood smear images under light microscope. All dataset images are captured from Hematology Unit, UKM Medical Centre in Kuala Lumpur. On all tested images, the cells of interest are successfully detected and counted from those clump cells. A comparative study and analysis to evaluate the performance of the proposed algorithm in three levels of clump have been conducted. The first level was with two clumps, second level with three clumps and third level with four clumps. The counting number of clump cells has been analyzed using quantitative analysis, resulting in much better results compared to other recent algorithms. The comparison shows that the proposed method gives better precision result at all levels with respect to ground truth: two clump cells (92%), three clump cells (96%) and four clump cells (90%). The results prove that this study has successfully developed a new method to count heavily clump RBC more accurately in microscopic images. In addition, this can be considered as a low-cost solution for quantification in massive examination

Detection of haemoglobin S using multiplex ligation-dependent probe amplification and flow-through hybridization techniques: experience in a tertiary hospital

Haemoglobin S (HbS, α2β26GluVal) is a variant haemoglobin resulted from GAGGTG mutation on codon 6 of HBB gene. HbS haemoglobinopathy is uncommon in Malaysia and mainly seen in immigrants. However, Malaysian Indians and Malays are rarely affected. This study reviewed the laboratory findings of patients with HbS and the utilization of multiplex ligation-dependent probe amplification (MLPA) and flow-through hybridization (FTH) in the detection of HbS mutation. HbS was identified and quantified by high performance liquid chromatography (HPLC), capillary electrophoresis (CE) and cellulose acetate gel electrophoresis. Molecular analysis was performed using MLPA, FTH and Sanger Sequencing. Two Africans, three Malays and two Indian individuals aged between 2-31 years were identified from our laboratory. Five patients were homozygous HbS, one was compound heterozygous HbS/β-thalassemia and one was a carrier of HbS. The patients with homozygous HbS had their haemoglobin (Hb) ranging from 7.4-10.2 g/dL with HbS and HbF levels of 58.3-94.7% and 1.5-35.5%, respectively. The Hb of compound heterozygous HbS/β-thalassaemia patients was 5.8 g/dL and was normal in heterozygous HbS. HbS, HbF and HbA2 levels for the HbS/β-thalassaemia and the carrier were 67%, 27.2% and 4.2%, and 38.6%, 0.1% and 2.8%, respectively. Both MLPA and FTH successfully detected HbS mutation in all cases but only FTH could identify the zygosity of the HbS mutation together with underlying concomitant β-thalassaemia in a single test

Detection of BCR-ABL T315i mutation in imatinib resistant chronic myeloid leukemia patients

Chronic myeloid leukemia (CML) patients who have BCR-ABL T315I mutation, usually present in the advance phase of the disease with overall survival (OS) shorter than those without the mutation. This study aimed to determine the prevalence of T315I mutation amongst imatinib mesylate (IM) resistant CML patients and to compare the OS between T315I-mutated and non-T315I-mutated patients. Sixty consecutive CML patients who were treated with IM for at least 18 months and their treatment responses, were recorded. The mutation analysis was done using allele-specific oligonucleotide reverse transcriptase-polymerase chain reaction (RT-PCR) assay followed by direct sequencing technique. Forty-two patients (70%) were found to have IM-resistance. Five out of 42 patients had detectable T315I mutation. Median OS of IM-resistant T315I-mutated patients was 96 months (95% CI:54-138) compared to 84 months (95% CI:48-120) in non T315I-mutated patients, although this was found to be statistically insignificant (p = 0.43). The present study showed a higher prevalence of T315I mutation as compared to a few local studies. Median OS of T315I-mutated patients were observed to be longer than non-T315-mutated patients. Further studies encompassing larger cohort of patients are required to confirm this finding

First reported case of haemoglobin-M Hyde Park in a Malay family living in Malaysia

Haemoglobin (Hb)-M Hyde Park, also known as Hb-M Akita is a rare type of hereditary Hb M due to autosomal dominant mutation of CAC>TAC on codon 92 of β globin gene resulting in the replacement of histidine by tyrosine on β globin chain. This variant Hb has a tendency to form methaemoglobin (metHb). The iron ion in metHb is oxidized to ferric (Fe3+) which is unable to carry oxygen and the patients manifest as cyanosis clinically. A 9-year-old Malay girl was incidentally found to be cyanotic when she presented to a health clinic. Laboratory investigations revealed raised methaemoglobin levels and Hb analysis findings were consistent with Hb-M Hyde Park. β gene sequencing confirmed a point mutation of CAC>TAC on codon 92 in one of the β genes. The family study done on the individuals with cyanosis showed similar findings. A diagnosis of heterozygous Hb-M Hyde Park was made. Patients with this variant Hb usually presented with cyanosis with mild haemolysis and maybe misdiagnosed as congenital heart disease. No further treatment is needed as patients are relatively asymptomatic. Although the disease is harmless in the heterozygous carriers but the offspring of the carriers may suffer severe haemolytic anaemia when the offspring also inherit other β haemoglobinopathies/thalassemia. This can happen due to high prevalence of β thalassemia carrier (3.5-4 %) found in Malaysia. At the time of writing, this is the first case of hereditary Hb-M Hyde Park diagnosed in a Malay family living in Malaysia

Primary Bone Marrow Lymphoma with Secondary Central Nervous System Involvement: a case report

Primary bone marrow lymphoma (PBML) is a rare condition. Most PBMLs are B-cell non-Hodgkin lymphomas (NHLs), where predominated by the diffuse large B-cell lymphomas (DLBCLs). Non-Hodgkin lymphomas affects extranodal sites in one-third of cases. Secondary central nervous system lymphoma (SCNSL) is a rare state which is defined as secondary central nervous system (CNS) involvement in patients with systemic lymphoma. In this report, we described a case of primary bone marrow lymphoma with secondary involvement of the CNS. Patient presented with neurological deficit. Peripheral blood film showed presence of abnormal mononuclear cells. Histopathological examination of the brain tissue showed features of DLBCL with CD10 positivity. Bone marrow examination showed presence of lymphoma cells. He was commenced with Rituximab, methotrexate, vincristine and procarbazine (R-MPV). However, he succumbed after two cycles of chemotherapy

Generation of a novel ex-vivo model to study re-endothelialization

AbstractEndothelial dysfunction initiates the pathogenesis of a myriad of cardiovascular diseases, yet the precise underlying mechanisms remain unclear. Current model utilises mechanical denudation of arteries resulting in an arterial-injury model with onset of intimal hyperplasia (IH). Our study shows that 5 min enzymatic denudation of human umbilical artery (hUA) lumen at 37 °C efficiently denudes hUA while maintaining vessel integrity without significantly increase intima-media thickness after 7 days in culture. This ex-vivo model will be a valuable tool in understanding the mechanism of re-endothelialization prior to smooth muscle cells (SMC) activation thus placating IH at an early stage

Conditions for polymerase chain reaction (PCR) technique.

Conditions for polymerase chain reaction (PCR) technique.</p

Prevalence and distribution of G6PD deficiency among the Senoi Malaysian Orang Asli population.

Prevalence and distribution of G6PD deficiency among the Senoi Malaysian Orang Asli population.</p

G6PD activity distributions (% Normal) for Coimbra, Kaiping, Union, Viangchan, and unknown variants among the Senoi Malaysian Orang Asli population.

G6PD activity distributions (% Normal) for Coimbra, Kaiping, Union, Viangchan, and unknown variants among the Senoi Malaysian Orang Asli population.</p