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The aim of this study was to examine the antioxidant capacity of Trypsin-hydrolyzed corn silk proteins, specifically radical scavenging, ferric reducing and anti-lipid peroxidation activities, as well as stability after heating and simulated gastrointestinal digestion. Among the 1–5-h hydrolysates, the 1-h Trypsin hydrolysate (T1H) was the strongest. T1H exhibited stronger H2O2 [half maximal effective concentration (EC50) = 156.44 µg/mL] and superoxide (EC50 = 0.33 mg/mL) scavenging activities than glutathione, carnosine and N-acetylcysteine. Radical scavenging activities of T1H were heat-stable (25–100°C), although lost by 20–58% after gastrointestinal digestion. Ferric reducing activity of T1H was heat-stable, even enhanced by 20% after gastrointestinal digestion. Lecithin liposome assay found anti-lipid peroxidation activity of T1H resistant to gastrointestinal peptidases. By contrast, 100°C incubation reduced anti-lipid peroxidation activity of 0.5 mg/mL T1H to 3.5-fold lower than that of 25°C incubation. At 30 µg/mL, T1H (3% hemolysis) was stronger than glutathione (32% hemolysis) in protecting human red blood cells. T1H was 1.5-fold more potent than glutathione and carnosine in alleviating lipid peroxidation in the cells. Overall, T1H demonstrated strong, heat-stable radical scavenging activities, besides partial resistance to gastrointestinal peptidases. T1H was also hematoprotective. T1H is a promising antioxidant for the development of functional food ingredients and nutraceuticals.

This study was conducted to identify and characterize antioxidant peptides from the alcalase hydrolysate of the blue-spotted stingray. Purification steps guided by ABTS cation radical (ABTS +) scavenging assay and de novo peptide sequencing produced two peptides, WAFAPA (661.3224 Da) and MYPGLA (650.3098 Da). WAFAPA (EC50 = 12.6 µM) had stronger antioxidant activity than glutathione (EC50 = 13.7 µM) and MYPGLA (EC50 = 19.8 µM). Synergism between WAFAPA and MYPGLA was detected. WAFAPA and MYPGLA surpassed carnosine in their ability to suppress H2O2-induced lipid oxidation. The peptides protected plasmid DNA and proteins from Fenton’s reagent-induced oxidative damage. Thermal (25–100 °C) and pH 3–11 treatments did not alter antioxidant activity of the peptides. MYPGLA maintained its antioxidant activity after simulated gastrointestinal digestion, whereas WAFAPA showed a partial loss. The two peptides may have potential applications as functional food ingredients or nutraceuticals, whether used singly or in combination.

Globally, peptide-based anticancer therapies have drawn much attention. Marine organisms are a reservoir of anticancer peptides that await discovery. In this study, we aimed to identify cytotoxic oligopeptides from Sarcophyton glaucum. Peptides were purified from among the S. glaucum hydrolysates produced by alcalase, chymotrypsin, papain, and trypsin, guided by a methylthiazolyldiphenyl-tetrazolium bromide (MTT) assay on the human cervical cancer (HeLa) cell line for cytotoxicity evaluation. Purification techniques adopted were membrane ultrafiltration, gel filtration chromatography, solid phase extraction (SPE), and reversed-phase high-performance liquid chromatography (RP-HPLC). Purified peptides were identified by de novo peptide sequencing. From papain hydrolysate, three peptide sequences were identified: AGAPGG, AERQ, and RDTQ (428.45, 502.53, and 518.53 Da, respectively). Peptides synthesized from these sequences exhibited cytotoxicity on HeLa cells with median effect concentration (EC50) values of 8.6, 4.9, and 5.6 mmol/L, respectively, up to 5.8-fold stronger than the anticancer drug 5-fluorouracil. When tested at their respective EC50, AGAPGG, AERQ, and RDTQ showed only 16%, 25%, and 11% cytotoxicity to non-cancerous Hek293 cells, respectively. In conclusion, AERQ, AGAPGG, and RDTQ are promising candidates for future development as peptide-based anticancer drugs.

Introduction: Alpha thalassaemia is one of the haemoglobin disorders in which the carriers of alpha thalassaemia may have normal haemoglobin level and are eligible to donate blood which may bring complications. This study is to investigate the interaction of haematological parameter with α-globin genotypes among eligible blood donors. Materials & Methods: A cohort study with 270 eligible blood donors were analysed for red cell indices. Alpha-globin (α-globin) genotyping was performed for seven deletions, six point mutations and two triplications. Statistical analyses were performed to compare the α-globin genotypes with haematological data. Results: High prevalence of α-thalassaemia carriers (7.7%, 21/270) was found among blood donors. All of them did not show anaemic pictures with a normal Hb level (>12 gm/dl). Five genotypes were identified consisting of 249 αα/αα (92.2%), nine -α 3.7/αα (3.3%), nine-- SEA/αα (3.3%), two -α 4.2/αα (0.7%) and one ααCS/αα (0.4%). Different α-globin genotypes showed a significant difference in RBC, MCV, MCH, MCHC, RDW, and Hct/Hb ratio (p

Stroke is the second leading cause of death and a major cause of disability worldwide. Both modifiable and non-modifiable risk factors can affect the occurrence of ischemic stroke at varying degrees. Among them, atherosclerosis has been well-recognized as one of the main culprits for the rising incidence of stroke-related mortality. Hence, the current review aimed to summarize the prominent role of lipid metabolism genes such as PCSK9, ApoB, ApoA5, ApoC3, ApoE, and ABCA1 in mediating ischemic stroke occurrence.

Objective The Filipino β°-deletion has been reported as a unique mutation in East Malaysia with a severe phenotype due to the complete absence of β-globin chain synthesis. In this study, the haplotype patterns of the β-globin gene cluster were used to relate the human genetic variation to this specific β-thalassaemia mutation. Methods The 376 study subjects included 219 β-thalassaemia major (β-TM) patients with homozygous Filipino β°-deletion and 157 carriers with heterozygous Filipino β°-deletion from 10 government hospitals in different regions of Sabah. Genomic DNA was isolated from whole blood using silica membrane based DNA purification protocol. Polymerase chain reaction restriction fragment length polymorphism analysis (PCR-RFLP) was conducted on five markers within the β-globin gene cluster to construct the haplotype patterns. Results Four haplotypes (Haplotype I-IV) were identified with Haplotype I as the predominant haplotype with the highest frequency of 0.98, followed by Haplotype II, III and Haplotype IV with 0.02. Haplotype I was strongly linked with the Filipino β°-deletion among the indigenous population. Conclusion Haplotype I as the predominant haplotype suggests the patients with the Filipino β°-deletion in Sabah have a similar origin.

Aging, which affects most of the multi-cellular organisms, is due to a potentially complex set of mechanisms that collectively cause a time-dependent decline of physiological functions. Aging restrains longevity and leads to neurodegenerative diseases including dementia, Alzheimer's disease and lacunar stroke. Human microbiota is now considered to have a strong impact on the progression of aging. The impact of aging and the risk of neurodegenerative diseases can be reduced by using probiotics, or preferably by combining probiotics and prebiotics, also known as synbiotics, that can drastically modify the composition of gut microbiome.

Fruits, vegetables, and herbs consist of various phytochemicals. Phytochemicals are proven to have beneficial effects for human health. It can be released from the plant materials into water by immersing them for a certain period of time, in which this method is widely known as water infusion. Common plant materials to be used for water infusion preparation are fruits such as lemon, apple, strawberry, grape, and pineapple and vegetables such as cucumber, okras, and many other herbs. Phytochemical compounds that can be extracted using water from plants include flavonoids (naringenin, quercetin, kaempferol, apigenin), phenolic compounds (curcumin, chlorogenic acid, gallic acid, hydroxybenzoic acid), carotenoids (β-carotene, lycopene, lutein), alkaloids (theobromine), and so on. A wide range of therapeutic properties, including antioxidant, antidiabetic, anti-inflammatory, antihypertension, anticancer, and antimicrobial can be benefited upon consumption of plant infused water as preferred healthy drinks for daily usage. Thus, infused water is an alternative natural way of treating or preventing illness without any side effect unlike other commercialized medications. Therefore, the present chapter deals with phytochemicals that can be extracted from fruits, vegetables, and herbs as well as emphasizing the health benefits derived from consumption of this plants infused water as potential preferred healthy drinks.

Purpose Filipino β0-deletion is a rare extensive deletion of β-globin gene, which predominantly found in β-thalassaemia patients in the indigenous population of Sabah, Malaysia particularly Kadazandusuns. Co-inherited of Filipino β0-deletion and −α3.7 deletion was commonly found among Kadazandusuns with milder clinical symptoms. Additionally, HBS1L-MYB, an important key regulator in haematopoiesis and erythropoiesis is strongly associated with the variation of HbF levels with milder clinical manifestation. Thus, this study investigates the influence of genetic variants in HBS1L-MYB on Hb subtypes level among thalassaemia carriers co-inherited with Filipino β°-deletion and −α3.7 deletion. Methods Gap polymerase chain reactions (gap-PCRs) were conducted to identify Filipino β0-deletion and −α3.7 deletion. Hb subtypes levels were quantified using BioRad Variant II Hb analyser. Two HBS1L-MYB intergenic polymorphisms (HMIPs)(rs9399137 and rs11759553) were genotyped using in house designed tetra primer ARMS-PCR. Confirmation of HMIPs were done through sequencing. Results Hundred and fifty-seven Filipino β°-deletion carriers from the Kadazandusun were recruited for this study. Out of 157 carriers, thirty-four Filipino β°-deletion thalassaemia carriers were detected for co-inheritance with −α3.7 deletion. The minor allele frequencies (MAF) of the two HMIPs were found more than 0.05 (rs11759553, MAF = 0.18 and rs9399137, MAF = 0.15). For rs11759553, genotypes AT and TT were found with significantly higher HbF level (p = .001). For rs9399137, C allele was found with higher HbF level but no significant difference between different genotypes. Both variants did not influence on the HbA and A2 level. Conclusions Both HMIPs rs11759553 and rs9399137 were significant polymorphisms among thalassaemia carriers co-inherited with Filipino β°-deletion and −α3.7 deletion carriers. Both HMIPs demonstrate higher expression of HbF level.

Introduction: Alpha thalassaemia is a highly prevalent disease globally and is a well-known public health problem in Malaysia. The deletional forms of the mutation are the most common forms found in alpha thalassaemia. The three most common deletional alpha thalassaemia found in this region include --SEA deletion, -α3.7 rightward and -α4.2 leftward deletions. The prevalence rate of triplication alpha cases such as αααanti3.7 and αααanti4.2 is not known in Malaysia although it plays a role in exacerbating the clinical phenotypes in beta thalassaemia carriers. Recently, there have been more reported cases of rare alpha thalassaemia mutations due to the advancement of molecular techniques involved in thalassaemia detections. Therefore, it is essential to develop a new method which allows the detection of different alpha thalassaemia mutations including the rare ones simultaneously and accurately. Methods: The purpose of this study was to design an assay for the detection of triplications, common and rare deletional alpha thalassaemia using droplet digital PCR (ddPCR). Results: This is a quantitative detection method to measure the changes of copy number which can detect deletions, duplications and triplications of the alpha globin gene simultaneously. Conclusion: In conclusion, ddPCR is an alternative method for rapid detection of alpha thalassaemia variants in Malaysia.

The diverse clinical phenotype of hemoglobin E (HbE)/β-thalassemia has not only confounded clinicians in matters of patient management but has also led scientists to investigate the complex mechanisms involved in maintaining the delicate red cell environment where, even with apparent similarities of α- and β-globin genotypes, the phenotype tells a different story. The BTB and CNC homology 1 (BACH1) protein is known to regulate α- and β-globin gene transcriptions during the terminal differentiation of erythroid cells. With the mutations involved in HbE/β-thalassemia disorder, we studied the role of BACH1 in compensating for the globin chain imbalance, albeit for fine-tuning purposes.A total of 47 HbE/β-thalassemia samples were analyzed using real-time quantitative polymerase chain reaction and correlated with age, sex, red blood cell parameters, globin gene expressions, and some clinical data.The BACH1 expression among the β-thalassemia intermedia patients varied by up to 2-log differences and was positively correlated to age; α-, β-, and γ-globin gene expression level; and heme oxygenase 1 protein. BACH1 was also negatively correlated to reticulocyte level and had a significant correlation with splenectomy.This study indicates that the expression of BACH1 could be elevated as a compensatory mechanism to decrease the globin chain imbalance as well as to reduce the oxidative stress found in HbE/β-thalassemia.Amaç: Hemoglobin E (HbE)/β-talaseminin çeşitli klinik fenotipleri klinisyenlerin hasta yönetimi esnasında zihinlerini karıştırmakla kalmamış, α- ve β-globin genotiplerinde bariz benzerlikler varken fenotiplerde farklılıklar bulunduğundan bilim insanlarının hassas eritrosit çevrenin muhafaza edilmesinde yer alan karmaşık mekanizmaları incelemelerine de ön ayak olmuştur. BTB ve CNC homoloji 1 (BACH1) proteininin eritroid hücrelerin son farklılaşması sırasında α- and β-globin gen transkripsiyonlarını ayarladığı bilinmektedir. HbE/β-talasemi hastalığındaki mutasyonlar ile her ne kadar ince ayar amaçlı ise de BACH1’in globin zincir dengesizliğini kompanse etmedeki rolünü inceledik. Gereç ve Yöntemler: Toplam 47 HbE/β-talasemi örneği gerçek zamanlı kantitatif polimeraz zincir reaksiyonu ile incelendi ve yaş, cinsiyet, eritrosit değişkenleri, globin gen sunumları ve bazı klinik veriler ile korele edildi. Bulgular: β-talasemi intermedia hastalarındaki BACH1 sunumu 2-log’a kadar farklılık göstermekteydi ve yaş; α-, β- ve γ-globin gen sunum düzeyleri; ve hem oksijenaz 1 proteini ile pozitif korelasyonu vardı. Ayrıca BACH1’in retikülosit düzeyi ile negatif korelasyonu vardı ve splenektomi ile anlamlı korelasyonu bulunmaktaydı. Sonuç: Bu çalışma hem HbE/β-talasemide bulunan oksidatif stresi hem de globin zincir dengesizliğini azaltmak için BACH1 sunumunun kompansasyon mekanizması olarak artabileceğini göstermiştir.

Purpose Obesity is still an issue worldwide with a high prevalence rate in spite of countless health awareness campaigns being held by both government and non-profit organisations. This issue is also faced in Malaysia. Thus, this is indeed to study on the risk factor of obesity, especially at the genetic aspect of eating behaviour which is still lacking and inconclusive. This study was to investigate the influence of dopamine type 2 receptors (DRD2) gene variants (ANKK1/DRD2 Taq1A, DRD2 Taq1B and DRD2 Taq1D) on eating behaviours [cognitive restraint eating (CR), emotional eating (EE) and uncontrolled eating (UE)] as well as to obesity. Methods The study consisted of 394 subjects (125 males and 269 females) who studied or worked in University Tunku Abdul Rahman (UTAR), Malaysia. Eating behaviour scores were assessed by using the Three-Factor Eating Questionnaire-R18 (TFEQ-R18). Body mass index (BMI) of the participants was taken to evaluate obesity status. Genotyping of Taq1 gene polymorphisms was carried out using Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) using the genomic DNAs extracted from mouthwash samples. Results There is no significant association between DRD2 gene variants with obesity (p > .05). However, ANKK1/DRD2 Taq1A gene polymorphism demonstrated a significant difference in EE score (p .05). Conclusions This study postulated that ANKK1/DRD2 Taq1A gene variant might have an influential effect on emotional eating among Malaysian adults. Besides, findings demonstrated no association of dopamine receptor gene variants with obesity. However, the role of eating behaviour in leading to obesity was yet inconclusive, and further study is required.

Fetal hemoglobin (HbF) is the predominant hemoglobin in red cells during fetal life. Just after birth, the level of HbF decreases gradually to1%, and is replaced mainly by adult hemoglobin (HbA) (∼ 97%). However, higher HbF levels could be associated with HbE/β-thalassemia, a complex thalassemia intermedia with a diverse clinical severity ranging from mild-to-severe anemia. This study investigates the correlation of HbF level with the clinical and laboratory data of HbE/β-thalassemia individuals.Peripheral blood samples from 30 HbE/β-thalassemia subjects were subjected to a full blood count, genomic as well as quantitative real-time polymerase chain reaction gene expression studies. Statistical analyses were performed using SPSS 17.0.HbF levels were influenced by age, mean cell volume (MCV), mean cell hemoglobin (MCH), HbA, β-globin, and α/β-globin expressions.HbF production is affected by the α/β-globin chain imbalance due to the lack of β-globin gene expression as well as inversely correlates to the amount of functional hemoglobin available in the cells.

Hypertension is a silent killer worldwide, caused by both genetic and environmental factors. Until now, genetic and genomic association studies of hypertension are reporting different degree of association on hypertension. Hence, it is essential to gather all the available information on the reported genetic loci and to determine if any biomarker(s) is/are significantly associated with hypertension. Current review concluded the potential biomarkers for hypertension, with regards to electrolyte and fluid transports, as well as sodium/potassium ions homeostasis, which are supported by the results of case-controls and meta-analyses.

Beta-thalassemia is one of the most prevalent inherited diseases and a public health problem in Malaysia. Malaysia is geographically divided into West and East Malaysia. In Sabah, a state in East Malaysia, there are over 1000 estimated cases of β-thalassemia major patients. Accurate population frequency data of the molecular basis of β-thalassemia major are needed for planning its control in the high-risk population of Sabah. Characterization of β-globin gene defects was done in 252 transfusion dependent β-thalassemia patients incorporating few PCR techniques. The study demonstrates that β-thalassemia mutations inherited are ethnically dependent. It is important to note that 86.9% of transfusion-dependent β-thalassemia major patients in Sabah were of the indigenous population and homozygous for a single mutation. The Filipino β(0)-deletion was a unique mutation found in the indigenous population of Sabah. Mutations common in West Malaysia were found in 11 (4.3%) patients. Four rare mutations (Hb Monroe, CD 8/9, CD 123/124/125 and IVS I-2) were also found. This study is informative on the population genetics of β-thalassemia major in Sabah.

Summary Aims Classical carriers of β 0 -thalassaemia are identified by a raised HbA 2 level. Earlier studies indicated that the Filipino β 0 -deletion has high raised HbA 2 levels. The introduction of automated high performance liquid chromatography (HPLC) for thalassaemia screening is an important advance in technology for haematology laboratories. The BioRad Variant II Hb analyser is a common instrument used to quantify HbA 2 levels in thalassaemia screening. This study aimed to determine HbA 2 levels in carriers of Filipino β 0 -mutation using the BioRad Variant II Hb analyser. Methods The Filipino β 0 -deletion was identified using gap-polymerase chain reaction (PCR) in the parents of transfusion dependent β 0 -thalassaemia patients who were homozygous for the Filipino β 0 -deletion in the indigenous population of Sabah, Malaysia. Hb subtypes were quantified on the BioRad Variant II Hb analyser. Concurrent α-thalassaemia was identified by multiplex gap-PCR for deletions and amplification refractory mutation system (ARMS)-PCR for non-deletional mutations. Results The mean HbA 2 level for Filipino β 0 -thalassaemia trait was 5.9 ± 0.47 and with coinheritance of α-thalassaemia was 6.3 ± 0.44 (-α 3.7 heterozygous) and 6.7 ± 0.36 (-α 37 homozygous). The HbA 2 levels were all >4% in keeping with the findings of classical β-thalassaemia trait and significantly higher than levels seen in non-deletional forms of β-thalassaemia. Conclusion The HbA 2 level measured on the BioRad Variant II Hb analyser was lower than the level in the first description of the Filipino β 0 -thalassaemia. β-thalassaemia trait with coinheritance of α-thalassaemia (-α 3.7 ) is associated with significantly higher HbA 2 level.

Hemoglobin (Hb) Adana [HBA2: c179G>A (or HBA1); p.Gly60Asp] is a non-deletional α-thalassemia variant found in Malaysia. An improvement in the molecular techniques in recent years has made identification of Hb Adana much easier. For this study, a total of 26 Hb Adana α-thalassemia intermedia and 10 Hb Adana trait blood samples were collected from patients. Common deletional and non-deletional α-thalassemia genotypes were determined using multiplex gap polymerase chain reaction (PCR) and multiplex ARMS PCR techniques. Identification of the Hb Adana location on the α-globin gene was carried out using genomic sequencing and the location of the mutation was confirmed via restriction fragment length polymorphism-PCR. Among the 36 samples, 24 (66.7%) had the -α(3.7)/α(Cd59)α mutation, while the -α(3.7)/α(Cd59)α mutation accounted for 2 samples (5.6%) and the remaining 10 (27.8%) samples were α/α(Cd59)α. All 36 samples were found to have the Hb Adana mutation on the α2-globin gene. The position of the α-globin gene mutation found in our cases was similar to that reported in Indonesia (16%) but not to that in Turkey (0.6%). Our results showed that the Hb Adana mutation was preferentially present in the α2-globin genes in Malays compared to the other ethnicities in Malaysia. Thus, the Malays might have similar ancestry based on the similarities in the Hb Adana position.

Hypertension is a silent killer worldwide, caused by both genetic and environmental factors. Until now, genetic and genomic association studies of hypertension are reporting different degree of association on hypertension. Hence, it is essential to gather all the available information on the reported genetic loci and to determine if any biomarker(s) is/are significantly associated with hypertension. Current review concluded the potential biomarkers for hypertension, with regards to electrolyte and fluid transports, as well as sodium/potassium ions homeostasis, which are supported by the results of case-controls and meta-analyses.

Summary Introduction In Malaysia, β-thalassaemia is a common inherited blood disorder in haemoglobin synthesis with a carrier rate of 4.5%. Currently, PCR-incorporating techniques such as amplification refractory mutation system (ARMS) or reverse dot blot hybridization (RDBH) are used in β-thalassaemia mutation detection. ARMS allows single-mutation identification using two reactions, one for wild type and another for mutant alleles. RDBH requires probe immobilization and optimization of hybridization and washing temperatures which is time consuming. The aim of our study was to investigate whether β-thalassaemia mutations can be identified in samples with low DNA concentrations. Methods Genotype identification of common β-thalassaemia mutations in Malays was carried out using Taqman genotyping assays. Results Results show that the Taqman assays allow mutation detection with DNA template concentrations as low as 2–100 ng. In addition, consistent reproducibility was observed in the Taqman assays when repeated eight times and at different time intervals. Conclusion The developed sensitive Taqman assays allow molecular characterization of β-thalassaemia mutations in samples with low DNA concentrations. The Taqman genotyping assays have potential as a diagnostic tool for foetal blood, chorionic villi or pre-implantation genetic diagnosis where DNA is limited and precious.

Detection and quantification of Hb subtypes of human blood is integral to presumptive identification of thalassaemias. It has been used in neonatal screening of thalassaemia and Hb variants. The use of discarded red blood cells following processing of the cord blood for stem cells provides readily available diagnostic material for thalassaemia screening. In this study, we determined the range of Hb subtypes in 195 consecutive cord blood samples collected for cord blood banking. The 'cord blood samples' analysed were those of the remaining red blood cells after the cord blood was processed for stem cell storage. Quantification of Hb subtypes by high performance liquid chromatography (HPLC) was done on BioRad Variant II Hb testing system. Only 73 (36.5%) of the samples could be analyzed neat without dilution. With a 1:300 dilution with wash solution the acceptable area as recommended by the manufacturer for reading of a C-gram within the 1 to 3 million ranges were achieved in all. Eighteen (9%) 12 showed classical Hb Barts (y4) prerun peaks were confirmed by Sebia Hydrasys automated Hb gel electrophoresis and quantified by Sebia Capillarys 2 capillary electrophoresis. Only 1 (0.5%) was presumptively identified with HbH disease. Due to the limited number of samples no beta-thalassaemia major, Hb E beta-thalassaemia and Hb Barts hydrops fetalis were found. The HPLC assay was possible at a cost US$ 5 per sample and a turnover time of 10 samples per hour without technical difficulties. This study reports an effective and valuable protocol for thalassaemia screening in red blood cells which would otherwise be discarded during cord blood processing. Cord blood with severe and intermediate forms of thalassaemia can be preselected and not stored.

The alpha haemoglobin stabilising protein (AHSP) acts as a molecular chaperone for α-globin by stabilising nascent α-globin before transferring it to waiting free β-globin chains. Binding of AHSP to α-globin renders α-globin chemically inert whereby preventing it from precipitating and forming reactive oxygen species byproducts. The AHSP has been actively studied in the recent years, particularly in its relation to β-thalassaemia. Studies have shown that AHSP is a modifier in β-thalassaemia mice models. However, this relationship is less established in humans. Studies by some groups showed no correlation between the AHSP haplotypes and the severity of β-thalassaemia, whereas others have shown that certain AHSP haplotype could modify the phenotype of β-thalassaemia intermedia patients. We investigated the expression of AHSP in relation to selected demographic data, full blood count, HPLC results, HbE/β-thalassaemia genotype, Xmn-1 Gγ polymorphism, α-globin, β-globin and γ-globin expression. We found that AHSP expression was significantly correlated to mean cell haemoglobin level, HbF %, α-globin, β-globin and excess α-globin expression. We concluded that AHSP could be a secondary compensatory mechanism in red blood cells to counterbalance the excess α-globin chains in HbE/β-thalassaemia individuals.

Introduction: Dried blood spots (DBS) are currently the recommended sample collection method for newborn screening programmes in America. Early diagnosis of beta-thalassaemia screening is essential as it provides an added advantage especially in sickle cell disease. Beta-thalassaemia frequency is high in many poor countries, and the cost of using commercial DNA extraction kits can be prohibitive. Our study assessed three methods that use minimal reagents and materials to extract DNA from DBS for beta-thalassaemia identification. Methods: The methods assessed in this study were Tris-EDTA (TE) buffer-based method by Bereczky et al. (American Journal of Tropical Medicine and Hygiene 72, 2005, 249), NaCL/NaOH/Sodium dodecyl sulphate (SDS) method by Huang et al. (Human Genetics 84, 1990, 129) and NaOH method by Zhou et al. (Analytical Biochemistry 354, 2006, 159). Extracted DNA was amplified for three common beta-thalassaemia mutations in Malaysia. Results: Amplicons derived from TE buffer-based method were very faint and almost nonexistent while the NaCl/NaOH/SDS method did not produce any visible amplicons. The extraction using NaOH method produced visible bands that were comparable to the standard method using extraction kit. Conclusion: The NaOH method is a simple method that uses minimal equipment and reagents that make it labour- and cost-effective. This method could be adopted by poorer countries to extract DNA for beta-thalassaemia mutation characterization.

Beta-thalassaemia is the most common inherited disease in Malaysia with a carrier rate of 4.5%. The spectrum of β-globin mutations has been well documented in the major ethnic groups in West Malaysia. Sabah in East Malaysia has over 1000 cases of transfusion dependent β-thalassaemia major patients. The complete spectrum of β-thalassaemia mutations in Sabah is still unknown. This study was done to characterise these β-thalassaemia mutations incorporating PCR techniques. A total of 253 β-thalassaemia major patients were recruited in this study following institutional clearance and informed consent. Homozygosity of the Filipino β-thalassaemia deletion was found in 217 (85.8%) patients, comprising of 136 Kadazandusuns, 19 Rungus, 10 Murut, 9 Sungai, 4 Bajau, 2 Bisaya and 1 Kedayu. Compound heterozygosity of Filipino β-thalassaemia deletion was found in 24 (9.5%) patients. Mutations common in West Malaysia were found in 11 (4.34%) patients (6 Chinese, 2 Brunei, 1 Banjar, 1 Bajau and 1 Malay). This study indicates ethnicity is informative in assisting β-thalassaemia diagnosis in Sabah. The indigenous population in Sabah had a unique mutation, the Filipino deletion. This study provides a platform in developing a simple and accurate molecular diagnosis protocol for β-thalassaemia mutations identification in the indigenous population of Sabah.