Browsing by Author "Ainoon, O"
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Publication Detection of Partial G6PD Deficiency using OSMMR2000-D Kit with Hb Normalization(Univ Kebangsaan Malaysia, Fac Medicine, 2014) ;Azma, RZ ;Zubaidah, MS ;Azlin, I ;Hafiza, A ;Nurasyikin, Y ;Hidayati, SN ;Farisah, ARN ;Hamidah, HNAinoon, OGlucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common enzyme deficiency worldwide including Malaysia. Screening of cord blood for partial G6PD deficiency is important as they are also prone to develop acute haemolysis. In this study, we determined the prevalence of partial G6PD deficient in paediatric population aged 1 month-12 years and normal term female neonates using OSMMR-D kit with haemoglobin (Hb) normalization and compare it with florescence spot test (FST). A total of 236 children, aged between between 1 month-12 years and 614 normal term female neonates were recruited for this study. Determination of normal means for G6PD activity and; cut-off points for partial and severe deficiency were determined according to WHO Working Group (1989). Determination of prevalence for partial deficiency for both groups (female patient) was done using this enzyme assay kit and findings were compared with FST. In this study, 15.7% (18/115) female children were classified as partial G6PD deficient by quantitative enzyme method (G6PD activity: 4.23-5.26U/gHb). However, FST only detected 0.9% (1/115) with minimal G6PD activity. The prevalence of partial G6PD deficiency in female neonate group was 3.42% (21/614) by enzyme assay versus 0.49% (3/614) by FST. This study concluded that our routine screening method using FST was unable to diagnose female heterozygotes. We recommend using this quantitative enzyme assay method by OSMMR-D kit since it was more sensitive in detecting G6PD deficiency in female neonates compared to FST. - Some of the metrics are blocked by yourconsent settings
Publication Extended and stable gene expression via nucleofection of MIDGE construct into adult human marrow mesenchymal stromal cells(Springer, 2012) ;Mok, PL ;Cheong, SK ;Leong, CF ;Chua, KHAinoon, OHuman mesenchymal stromal cell (hMSC) is a potential target for cell and gene therapy-based approaches against a variety of different diseases. Whilst cationic lipofection has been widely experimented, the Nucleofector technology is a relatively new non-viral transfection method designed for primary cells and hard-to-transfect cell lines. Herein, we compared the efficiency and viability of nucleofection with cationic lipofection, and used the more efficient transfection method, nucleofection, to deliver a construct of minimalistic, immunologically defined gene expression encoding the erythropoietin (MIDGE-EPO) into hMSC. MIDGE construct is relatively safer than the viral and plasmid expression systems as the detrimental eukaryotic and prokaryotic gene and sequences have been eliminated. Using a plasmid encoding the luciferase gene, we demonstrated a high transfection efficiency using the U-23 (21.79 +/- 1.09%) and C-17 (5.62 +/- 1.09%) pulsing program in nucleofection. The cell viabilities were (44.93 +/- 10.10)% and (21.93 +/- 5.72)%, respectively 24 h post-nucleofection. On the other hand, lipofection treatment only yielded less than 0.6% efficiencies despite showing higher viabilities. Nucleofection did not affect hMSC renewability, immunophenotype and differentiation potentials. Subsequently, we nucleofected MIDGE-EPO using the U-23 pulsing program into hMSC. The results showed that, despite a low nucleofection efficiency with this construct, the EPO protein was stably expressed in the nucleofected cells up to 55 days when determined by ELISA or immunocytochemical staining. In conclusion, nucleofection is an efficient non-viral transfection approach for hMSC, which when used in conjunction with a MIDGE construct, could result in extended and stable transgene expression in hMSC. - Some of the metrics are blocked by yourconsent settings
Publication Human mesenchymal stromal cells could deliver erythropoietin and migrate to the basal layer of hair shaft when subcutaneously implanted in a murine model(Churchill Livingstone, 2012) ;Mok, PL ;Cheong, SK ;Leong, CF ;Chua, KHAinoon, OMesenchymal stromal cells (MSC) are an attractive cell-targeting vehicle for gene delivery. MIDGE (an acronym for Minimalistic, Immunologically Defined Gene Expression) construct is relatively safer than the viral or plasmid expression system as the detrimental eukaryotic and prokaryotic gene and sequences have been eliminated. The objective of this study was to test the ability of the human MSC (hMSC) to deliver the erythropoietin (EPO) gene in a nude mice model following nucleofection using a MIDGE construct. hMSC nucleofected with MIDGE encoding the EPO gene was injected subcutaneously in Matrigel at the dorsal flank of nude mice. Subcutaneous implantation of nucleofected hMSC resulted in increased hemoglobin level with presence of human EPO in the peripheral blood of the injected nude mice in the first two weeks post-implantation compared with the control groups. The basal layer of the hair shaft in the dermal layer was found to be significantly positive for immunohistochemical staining of a human EPO antibody. However, only a few basal layers of the hair shaft were found to be positively stained for CD105. In conclusion, hMSC harboring MIDGE-EPO could deliver and transiently express the EPO gene in the nude mice model. These cells could be localized to the hair follicle and secreted EPO protein might have possible role in hair regeneration. (C) 2012 Elsevier Ltd. All rights reserved. - Some of the metrics are blocked by yourconsent settings
Publication Molecular characteristic of alpha thalassaemia among patients diagnosed in UKM Medical Centre(Malaysian Journal Pathology, 2014) ;Azma, RZ ;Ainoon, O ;Hafiza, A ;Azlin, I ;Noor Farisah, AR ;Nor Hidayati, SNoor Hamidah, HAlpha (alpha) thalassaemia is the most common inherited disorder in Malaysia. The clinical severity is dependant on the number of alpha genes involved. Full blood count (FBC) and haemoglobin (Hb) analysis using either gel electrophoresis, high performance liquid chromatography (HPLC) or capillary zone electrophoresis (CE) are unable to detect definitively alpha thalassaemia carriers. Definitive diagnosis of alpha-thalassaemias requires molecular analysis and methods of detecting both common deletional and non-deletional molecular abnormailities are easily performed in any laboratory involved in molecular diagnostics. We carried out a retrospective analysis of 1623 cases referred to our laboratory in Universiti Kebangsaan Malaysia Medical Centre (UKMMC) for the diagnosis of alpha-thalassaemia during the period October 2001 to December 2012. We examined the frequency of different types of alpha gene abnormalities and their haematologic features. Molecular diagnosis was made using a combination of multiplex polymerase reaction (PCR) and real time PCR to detect deletional and non-deletional alpha genes relevant to southeast Asian population. Genetic analysis confirmed the diagnosis of alpha-thalassaemias in 736 cases. Majority of the cases were Chinese (53.1%) followed by Malays (44.2%), and Indians (2.7%). The most common gene abnormality was alpha alpha/--(SEA) (64.0%) followed by alpha alpha/-alpha(3.7) (19.8%), -alpha(3.7)/--(SEA) (6.9%), alpha alpha/alpha alpha(CS) (3.0%), --(SEA)/--(SEA) (1.2%), -alpha(3.7)/-alpha(3.7) (1.1%), alpha alpha/-alpha(4.2) (0.7%), -alpha(4.2)/--(SEA) (0.7%), -alpha(3.7)/-alpha(4.2) (0.5%), alpha alpha(CS)/--(SEA) (0.4%), alpha alpha(CS)/alpha alpha(Cd59) (0.4%), alpha alpha(CS)/alpha alpha(CS) (0.4%), -alpha(3.7)/alpha alpha(Cd59) (0.3%), alpha alpha/alpha alpha(Cd59) (0.1%), alpha alpha(Cd59)/alpha alpha(IVS) (I-1) (0.1%), -alpha(3.7)/alpha alpha(CS) (0.1%) and --(SEA)/alpha alpha(Cd59) (0.1%). This data indicates that the molecular abnormalities of alpha-thalassaemia in the Malaysian population is heterogenous. Although alpha-gene deletion is the most common cause, non-deletional alpha-gene abnormalities are not uncommon and at least 3 different mutations exist. Establishment of rapid and easy molecular techniques is important for definitive diagnosis of alpha thalassaemia, an important prerequisite for genetic counselling to prevent its deleterious complications. - Some of the metrics are blocked by yourconsent settings
Publication Prenatal diagnosis of aneuploidies in amniotic fluid by multiple ligation-dependent probe amplification (MLPA) analysis(Malaysian Journal Pathology, 2014) ;Hamidah, NH ;Munirah, AR ;Hafiza, A ;Farisah, AR ;Shuhaila, A ;Norzilawati, MN ;Jamil, MYAinoon, OPrenatal diagnosis is essential in the new era of diagnosis and management of genetic diseases in obstetrics. Multiple ligation-dependent probe amplification (MLPA) is a recent technique for prenatal diagnosis for the relative quantification of 40 different nucleic acid sequences in one single reaction. We had utilized the MLPA technique in detecting aneuploidies in amniotic fluid samples from 25 pregnant women from the Obstetrics and Gynaecology Department UKMMC, versus the quantitative fluorescent polymerase chain reaction (QF-PCR) method. Conclusive results were obtained in 18 cases and all were concordant with that of the QF-PCR. All four cases of trisomies were correctly identified including one case with maternal cell contamination.