MyMedR

Displaying all 9 publications

Abstract:

Sort:

Fulltext Isolation of nucleated red blood cell from peripheral blood of β-Thalassemia major patients using CD71 magnetic beads and future application

Haiyuni, M.Y., Aziee S., Heba A., Rosline H., Abdullah W.Z., Johan M.F., et al.

Journal of Biomedical and Clinical Sciences, 2018;3(1):25-30.
MyJurnal

Introduction: Isolation of specific cell types is important in providing a better understanding of hematological disorders. The knowledge of molecular biology aspect in β-thalassemia is still limited. This is because hemoglobin disorder involves various erythropoietic processes in which the genetic information is lack due to enucleation of red blood cells occurs in bone marrow. It is invasive to collect samples from bone marrow and cord blood although nucleated red blood cells (NRBCs) are abundant in these sites. NRBCs are precursors of red blood cells and typically found in peripheral blood (PB) of β-thalassemia major patients and abundant post-splenectomy. The utilization of PB NRBCs will provide a further understanding of the molecular aspects of ineffective erythropoiesis in β-thalassemia major patients. Objective: The objective of this study was to isolate the NRBCs using CD71 magnetic beads from PB of β-thalassemia major; non-splenectomy and post-splenectomy patients. Methods: NRBCs were isolated from 6 mL PB of β-thalassemia major patients based on density gradient and magnetic activated cell sorting (MACS) for NRBCs enrichment using a CD71 marker. Cell count was determined by using hemocytometer (Weber Scientific, NJ, USA) and BD FACSCantoTM II flow cytometry (Becton-Dickson, NJ, USA) was performed for method validation. Results: NRBCs were successfully isolated from the PB of both non-splenectomy and post-splenectomy β-thalassemia major patients with >90% specificity by flow cytometric analysis. The median number of enriched NRBCs (x104 ) was 58.5 (283) and 340 (338) respectively using hemocytometer. Conclusion: The MACS method was found to be convenient and efficient in the isolation of the targeted cells for downstream applications.
Fulltext School bag load and its effect on erector spinae muscle and low back pain among primary school children in Malaysia

Tamrin SBM, Ai LP, Hamzah R, Abdullah MY, Hanafi SS

Malaysian Journal of Medicine and Health Sciences, 2005;1(1):21-31.

Objective: A cross-sectional study was conducted to determine and evaluate the effects of schoolbag load on electromyography (EMG) activity of the erector spinae muscle.
Methods: Eighty-four primary schoolchildren were selected from two national medium primary schools in Seri Kembangan, 42 from Primary Two (P2) and 42 from Primary Five (PS), aged 8 and 11 respectively. Data were collected through interviews, anthropometries measurements, the weight of schoolbag load and surface electromyography (SEMG) of the erector spinae using Muscle Tester ME3000f'®. SurfaceAg-AgCl electrodes were used to measure: unloaded standing and walking, and loaded standing and walking.
Results: The study revealed that the erector spinae was found to be more efficient in loaded standing probably due to other trunk muscle co-activity compared to unloaded standing (p<0.05). However, the erector spinae was less efficient when loads were carried on the back while walking, compared to unloaded walking. When the schoolbag was carried over both shoulders, forces generated by the erector spinae were reduced and resulted in a more efficient use of the erector spinae compared to other asymmetrical carrying methods (p<0.05).
Conclusions: Bending slightly forward when carrying schoolbag was found to reduce the forces generated by the erector spinae compared to normal sagittal posture (p<0.05), however no significant difference was found between the different frontal postures. A significant inverse relationship (p<0.01) between the weight of schoolbag load and the average electromyography (AEMG), showed that the signifieance of the erector spinae muscle was reduced when a heavier schoolbag load was carried, owing to other trunk muscles co-activity. The study also revealed that apart from age of schoolchildren, family history of back pain, exposure to environmental tobacco smoke (ETS), weight of the schoolbag and method of carrying schoolbag also play important role as risk factors for back pain.
Keywords: Electromyography, schoolbag, primary school, back pain
Genetic polymorphisms of HbE/beta thalassemia related to clinical presentation: implications for clinical diversity

Azman NF, Abdullah WZ, Hanafi S, Diana R, Bahar R, Johan MF, et al.

Ann Hematol, 2020 Apr;99(4):729-735.
PMID: 32078010 DOI: 10.1007/s00277-020-03927-5

HbE/Beta thalassemia (HbE/β-thalassemia) is one of the common genetic disorders in South East Asia. It is heterogeneous in its clinical presentation and molecular defects. There are genetic modifiers which have been reported to influence the disease severity of this disorder. The aim of this study was to determine the genetic polymorphisms which were responsible for the disease clinical diversity. A case-control study was conducted among Malay transfusion-dependent HbE/β-thalassemia patients. Patients who were confirmed HbE/β-thalassemia were recruited and genotyping study was performed on these subjects. Ninety-eight patients were selected and divided into moderate and severe groups based on clinical parameters using Sripichai scoring system (based on hemoglobin level, spleen size, growth development, the age of first transfusion and age of disease presentation). Forty-three (44.9%) and 55 (56.1%) patients were found to have moderate and severe clinical presentation, respectively. Genotyping analysis was performed using Affymetrix 6.0 microarray platform. The SNPs were filtered using PLINK and Manhattan plot by R software. From the GWAS results, 20 most significant SNPs were selected based on disease severity when compared between moderate and severe groups. The significant SNPs found in this study were mostly related to thalassemia complications such as rs7372408, associated with KCNMB2-AS1 and SNPs associated with disease severity. These findings could be used as genetic predictors in managing patients with HbE/β-thalassemia and served as platform for future study.
Fulltext Prenatal auditory stimulation induces physiological stress responses in developing embryos and newly hatched chicks

Hanafi SA, Zulkifli I, Ramiah SK, Chung ELT, Kamil R, Awad EA

Poult Sci, 2023 Feb;102(2):102390.
PMID: 36608455 DOI: 10.1016/j.psj.2022.102390

Prenatal stress may evoke considerable physiological consequences on the developing poultry embryos and neonates. The present study aimed to determine prenatal auditory stimulation effects on serum levels of ceruloplasmin (CPN), alpha-1-acid glycoprotein (AGP), corticosterone (CORT), and heat shock protein 70 (Hsp70) regulations in developing chicken embryos and newly hatched chicks. Hatching eggs were subjected to the following auditory treatments; 1) control (no additional sound treatment other than the background sound of the incubator's compressors at 40 dB), 2) noise exposure (eggs were exposed to pre-recorded traffic noise at 90 dB) (NOISE), and 3) music exposure (eggs were exposed to Mozart's Sonata for Two Pianos in D Major, K 488 at 90 dB) (MUSIC). The NOISE and MUSIC treatments were for 20 min/h for 24 h (a total of 8 h/d), starting from embryonic days (ED) 12 to hatching. The MUSIC (1.37 ± 0.1 ng/mL) and NOISE (1.49 ± 0.2 ng/mL) treatments significantly elevated CPN at ED 15 compared to the Control (0.82 ± 0.04 ng/mL) group and post-hatch day 1 (Control, 1.86 ± 0.2 ng/mL; MUSIC, 2.84 ± 0.4 ng/mL; NOISE, 3.04 ± 0.3 ng/mL), AGP at ED 15 (Control, 39.1 ± 7.1 mg/mL; MUSIC, 85.5 ± 12.9 mg/mL; NOISE, 85.4 ± 15.1 mg/mL) and post-hatch day 1 (Control, 20.4 ± 2.2 mg/mL; MUSIC, 30.5 ± 4.7 mg/mL; NOISE, 30.3 ± 1.4 mg/mL). CORT significantly increased at ED 15 in both MUSIC (9.024 ± 1.4 ng/mL) and NOISE (12.15 ± 1.6 ng/mL) compared to the Control (4.39 ± 0.7 ng/mL) group. On the other hand, MUSIC exposed embryos had significantly higher Hsp70 expression than their Control and NOISE counterparts at ED 18 (Control, 12.9 ± 1.2 ng/mL; MUSIC, 129.6 ± 26.4 ng/mL; NOISE, 13.3 ± 2.3 ng/mL) and post-hatch day 1 (Control, 15.2 ± 1.7 ng/mL; MUSIC, 195.5 ± 68.5 ng/mL; NOISE, 13.2 ± 2.7 ng/mL). In conclusion, developing chicken embryos respond to auditory stimulation by altering CPN, AGP, CORT, and Hsp70. The alterations of these analytes could be important in developing embryos and newly hatched chicks to cope with stress attributed to auditory stimulation.
Prevalence and Distribution of Major β-Thalassemia Mutations and HbE/β-Thalassemia Variant in Nepalese Ethnic Groups

Lama R, Yusof W, Shrestha TR, Hanafi S, Bhattarai M, Hassan R, et al.

Hematol Oncol Stem Cell Ther, 2022 Mar 01;15(1):279-284.
PMID: 33592169 DOI: 10.1016/j.hemonc.2021.01.004

BACKGROUND: Beta-thalassemia is a genetic disorder that is inherited in an autosomal recessive pattern. This genetic disease leads to a defective beta-globin hemoglobin chain causing partial or complete beta-globin chain synthesis loss. Beta-thalassemia major patients need a continuous blood transfusion and iron chelation to maintain the normal homeostasis of red blood cells (RBCs) and other systems in the body. Patients also require treatment procedures that are costly and tedious, resulting in a serious health burden for developing nations such as Nepal.
METHODS: A total of 61 individuals clinically diagnosed to have thalassemia were genotyped with multiplex amplification refractory mutation system-polymerase chain reaction (ARMS-PCR). Twenty-one major mutations were investigated using allele-specific primers grouped into six different panels.
RESULTS: The most common mutations found (23%) were IVS 1-5 (G-C) and Cd 26 (G-A) (HbE), followed by 619 deletion, Cd 8/9 (+G), Cd 16 (-C), Cd 41/42 (-TTCT), IVS 1-1 (G-T), Cd 19 (A-G), and Cd 17 (A-T) at 20%, 12%, 8%, 6%, 4%, 3%, and 1%, respectively.
CONCLUSION: The results of this study revealed that Nepal's mutational profile is comparable to that of its neighboring countries, such as India and Myanmar. This study also showed that thalassemia could be detected across 17 Nepal's ethnic groups, especially those whose ancestors originated from India and Central Asia.
Fulltext Genotype-phenotype association of Hbe/β thalassemia disease and the role of genetic modifiers

HanafI S, Abdullah WZ, Adnan RA, Bahar R, Johan MF, Azman NF, et al.

Malaysian Journal of Paediatrics and Child Health, 2016;22(1):1-16.
MyJurnal

HbE/β-thalassemia is the most common severe form of thalassemia particularly in SEA region including Malaysia and globally, it comprised of a significant severe form of β-thalassemia disorder. It has various clinical manifestations ranging from very mild anemia to severe manifestation similar to beta thalassemia major. Many different syndromes are observed in HbE/β-thalassemia. Several genetic modifiers have been reported to play important role in contributing to phenotypic variability. The true reasons underlying this phenotypic variability remain unknown. The most reliable predictive factor of the disease phenotype is the nature of the beta globin gene mutation itself. However, the degree of severity is also believed to be affected by other genetic modifiers. For instance, high HbF level ameliorates the clinical severity of β thalassemia patients. Therefore, identification of these genetic modifiers is very important. The association of severe clinical manifestation and the specific β-globin gene mutation has been known. But the wide scope and other potential predictors have been only recently appreciated. This review therefore aimed to reveal the potential genetic modifiers of HbE/βthalassemia patients based on the previous reported studies. A better understanding on the mechanisms underlying the variety of phenotypes of this disease may lead to the direction for a better future management plans. This also promotes “personalized medicine” in patient care.
Fulltext Multiplex amplification refractory mutation system (MARMS) for the detection of β-globin gene mutations among the transfusion-dependent β-thalassemia Malay patients in Kelantan, Northeast of Peninsular Malaysia

Hanafi S, Hassan R, Bahar R, Abdullah WZ, Johan MF, Rashid ND, et al.

Am J Blood Res, 2014;4(1):33-40.
PMID: 25232503

The aim of this study was to adapt MARMS with some modifications to detect beta mutation in our cohort of thalassemia patients. We focused only on transfusion-dependent thalassemia Malay patients, the predominant ethnic group (95%) in the Kelantanese population. Eight mutations were identified in 46 out of 48 (95.83%) beta thalassemia alleles. Most of the patients (54.2%) were compound heterozygous with co-inheritance Cd 26 (G>A). The frequencies of spectrum beta chain mutation among these patients are presented in Table 2. Among the transfusion dependent beta thalassemia Malay patients studied, 26 patients were found to be compound heterozygous and the main alleles were Cd 26 (G>A). Compound heterozygous mutation of Cd 26 (G>A) and IVS 1-5 (G>C) were 12 (46.2%), Cd 26 (G>A) and Cd 41/42 (TTCT) were 9 (34.6%), Cd 26 (G>A) and IVS 1-1 (G>C) were 2 (7.7%) respectively. Meanwhile the minority were made of a single compound heterozygous of Cd 26 (G>A) and Cd 71/72, Cd 26 (>A) and Cd 17 (A>T), Cd 26 (G>A) and -28 (G>A) respectively. Twenty out of forty six patients were shown to have homozygous of IVS 1-5 (G>C) were 2 (10.0%), Cd 26 (G>A) were 15 (75.0%), Cd 19 (A>G) were 1 (5.0%), and IVS 1-1 (G>T) were 2 (10.0%). The beta chain mutations among the Kelantanese Malays followed closely the distribution of beta chain mutations among the Thais and the Malays of the Southern Thailand. The G-C transition at position 5 of the IVS 1-5 mutation was predominant among the Malay patients. In conclusion, this method has successfully identified the mutation spectrum in our cohort of transfusion-dependent beta thalassemia patients, and this method is equally effective in screening for mutation among thalassemia patients.
Fulltext Rapid Targeted Next-Generation Sequencing Platform for Molecular Screening and Clinical Genotyping in Subjects with Hemoglobinopathies

Shang X, Peng Z, Ye Y, Asan, Zhang X, Chen Y, et al.

EBioMedicine, 2017 Sep;23:150-159.
PMID: 28865746 DOI: 10.1016/j.ebiom.2017.08.015

Hemoglobinopathies are among the most common autosomal-recessive disorders worldwide. A comprehensive next-generation sequencing (NGS) test would greatly facilitate screening and diagnosis of these disorders. An NGS panel targeting the coding regions of hemoglobin genes and four modifier genes was designed. We validated the assay by using 2522 subjects affected with hemoglobinopathies and applied it to carrier testing in a cohort of 10,111 couples who were also screened through traditional methods. In the clinical genotyping analysis of 1182 β-thalassemia subjects, we identified a group of additional variants that can be used for accurate diagnosis. In the molecular screening analysis of the 10,111 couples, we detected 4180 individuals in total who carried 4840 mutant alleles, and identified 186 couples at risk of having affected offspring. 12.1% of the pathogenic or likely pathogenic variants identified by our NGS assay, which were undetectable by traditional methods. Compared with the traditional methods, our assay identified an additional at-risk 35 couples. We describe a comprehensive NGS-based test that offers advantages over the traditional screening/molecular testing methods. To our knowledge, this is among the first large-scale population study to systematically evaluate the application of an NGS technique in carrier screening and molecular diagnosis of hemoglobinopathies.
MortalitY in caRdIAc surgery (MYRIAD): A randomizeD controlled trial of volatile anesthetics. Rationale and design

Landoni G, Lomivorotov V, Pisano A, Nigro Neto C, Benedetto U, Biondi Zoccai G, et al.

Contemp Clin Trials, 2017 08;59:38-43.
PMID: 28533194 DOI: 10.1016/j.cct.2017.05.011

OBJECTIVE: There is initial evidence that the use of volatile anesthetics can reduce the postoperative release of cardiac troponin I, the need for inotropic support, and the number of patients requiring prolonged hospitalization following coronary artery bypass graft (CABG) surgery. Nevertheless, small randomized controlled trials have failed to demonstrate a survival advantage. Thus, whether volatile anesthetics improve the postoperative outcome of cardiac surgical patients remains uncertain. An adequately powered randomized controlled trial appears desirable.
DESIGN: Single blinded, international, multicenter randomized controlled trial with 1:1 allocation ratio.
SETTING: Tertiary and University hospitals.
INTERVENTIONS: Patients (n=10,600) undergoing coronary artery bypass graft will be randomized to receive either volatile anesthetic as part of the anesthetic plan, or total intravenous anesthesia.
MEASUREMENTS AND MAIN RESULTS: The primary end point of the study will be one-year mortality (any cause). Secondary endpoints will be 30-day mortality; 30-day death or non-fatal myocardial infarction (composite endpoint); cardiac mortality at 30day and at one year; incidence of hospital re-admission during the one year follow-up period and duration of intensive care unit, and hospital stay. The sample size is based on the hypothesis that volatile anesthetics will reduce 1-year unadjusted mortality from 3% to 2%, using a two-sided alpha error of 0.05, and a power of 0.9.
CONCLUSIONS: The trial will determine whether the simple intervention of adding a volatile anesthetic, an intervention that can be implemented by all anesthesiologists, can improve one-year survival in patients undergoing coronary artery bypass graft surgery.