42 results on '"Tan Jin Ai, Mary Anne"'
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2. Characterization of the relationship between polar and lateral flagellar genes in clinical Aeromonas dhakensis: phenotypic, genetic and biochemical analyses
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Lau, Tien-Tien Vicky, Puah, Suat-Moi, Tan, Jin-Ai Mary Anne, Puthucheary, S. D., and Chua, Kek-Heng
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- 2021
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3. Genetic relatedness and novel sequence types of clinical Aeromonas dhakensis from Malaysia
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Lau, Tien Tien Vicky, Tan, Jin-Ai Mary Anne, Puthucheary, S. D., Puah, Suat-Moi, and Chua, Kek-Heng
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- 2020
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4. Non-invasive Sampling for Assessment of Oxidative Stress and Pro-inflammatory Cytokine Levels in Beta-Thalassaemia Major Patients
- Author
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Rahim Mohd. Rashdan Abd., Tan Jin Ai Mary Anne, Mani Ravishankar Ram, and Kuppusamy Umah Rani
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β-thalassaemia major ,saliva ,oxidative stress ,biochemical analysis ,Medicine - Abstract
Background: Beta-thalassaemia (β-thalassaemia) major patients are severely anaemic and require life-long blood transfusions for survival. These patients require iron-chelation therapy as a result of iron overload due to the monthly blood transfusions. The iron over load can cause oxidative damage and pro-inflammation and therefore, hasten mortality. Thus, regular monitoring of the oxidative stress and pro-inflammation status may be useful in these patients.
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- 2016
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5. A holistic approach to education programs in thalassemia for a multi-ethnic population: consideration of perspectives, attitudes, and perceived needs
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Wong, Li Ping, George, Elizabeth, and Tan, Jin-Ai Mary Anne
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- 2011
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6. Interaction of Hb South Florida (codon 1; GTG→ATG) and HbE, with β-thalassemia (IVS1-1; G→A): expression of different clinical phenotypes
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Tan, Jin-Ai Mary Anne, Tan, Kim-Lian, Omar, Khairul Zaman, Chan, Lee-Lee, Wee, Yong-Chui, and George, Elizabeth
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- 2009
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7. 18q21 Rearrangement and trisomy 3 in extranodal B-cell lymphomas: a study using a fluorescent in situ hybridisation technique
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Tai, Yan-Chin, Tan, Jin-Ai Mary Anne, and Peh, Suat-Cheng
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- 2004
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8. Characterisation and confirmation of rare beta-thalassaemia mutations in the Malay, Chinese and Indian ethnic groups in Malaysia
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Tan, Jin Ai Mary Anne, Chin, Pui See, Wong, Yean Ching, Tan, Kim Lian, Chan, Lee Lee, and George, Elizabeth
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- 2006
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9. BamH1 polymorphism in the Chinese, Malays, and Indians in Singapore and its application in the prenatal diagnosis of β-thalassemia
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Tan, Jin Ai Mary Anne, Tay, Sin Hock John, Kham, Kow Yin Shirley, and Wong, Hock Boon
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- 1993
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10. Heterogeneity in α-thalassemia interactions in Malays, Chinese and Indians in Malaysia
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Wee, Yong-Chui, Tan, Kim-Lian, Chow, Teresa Wai-Ping, Yap, Sook-Fan, and Tan, Jin-Ai Mary Anne
- Published
- 2005
11. Higher frequency of p53 gene mutations in diffuse large B-cell lymphoma with MALT component
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Tai, Yan-Chin, Tan, Jin-Ai Mary Anne, and Peh, Suat-Cheng
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- 2004
12. Isolation, molecular characterization and antimicrobial susceptibility of Aeromonas spp. obtained from Tiger Grouper (Epinephelus fuscoguttatus) and Marble Goby (Oxyeleotris marmoratus) fish in Sabah, Malaysia.
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Lau, Tien‐Tien Vicky, Puah, Suat‐Moi, Hon, Ching‐Kee Karen, Ching, Fui‐Fui, Tan, Jin‐Ai Mary Anne, Puthucheary, Savithri Devi Ampalam, Lee, Ping‐Chin, and Chua, Kek‐Heng
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GOBIIDAE ,FISH farming ,EPINEPHELUS ,AEROMONAS ,FISHERIES ,FISHES - Abstract
Aeromonads are ubiquitous in aquatic environments and have been implicated in fish and human infections. In this study, we isolated, studied antimicrobial susceptibility patterns and screened the existence of 15 virulence genes in aeromonads from two famously consumed fish species—seven marine Tiger Grouper (Epinephelus fuscoguttatus) and eight freshwater Marble Goby (Oxyeleotris marmoratus) from the aquaculture hatchery in Sabah, Malaysia. A total of 30 aeromonads (17 A. caviae, 9 A. rivuli, 4 A. dhakensis) were identified using PCR targeting GCAT gene, rpoD‐restriction fragment length polymorphism and multi‐locus phylogenetic analysis. All 30 strains were resistant to amoxicillin and cephalothin and five strains were multidrug‐resistant. Nine virulence genes (lip, ela, eno, fla, aerA, hylA, dam, alt and ser) present in A. dhakensis, suggesting the virulence potential of this species as a fish pathogen. This study offers as a baseline for future studies in monitoring and managing these two fish in aquaculture industry. [ABSTRACT FROM AUTHOR]
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- 2020
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13. Isolation and molecular identification of Aeromonas species from the tank water of ornamental fishes.
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Cheok, Yi Ying, Puah, Suat Moi, Chua, Kek Heng, and Tan, Jin Ai Mary Anne
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ORNAMENTAL fishes ,AEROMONAS ,TANKS ,FRESHWATER fishes ,FISH pathogens ,RNA polymerases - Abstract
Aeromonads are recognised as important pathogens of fishes. In this study, ten water samples were randomly collected from pet shops' fish tanks and home aquaria inhabited by several fish species (silver arowana, koi, goldfish, catfish, pictus fish, silver shark and silver dollar fish). Altogether 298 colonies were isolated using Aeromonas selective agar. A total of 154 isolates were then confirmed as belonging to the genus Aeromonas using the GCAT gene. Using ERIC-PCR, a total of 40 duplicate isolates were excluded from the study and 114 isolates were subjected to PCR-RFLP targeting the RNA polymerase sigma factor (rpoD) gene using lab-on-chip. A total of 13 different Aeromonas species were identified. The most prevalent species were A. veronii (27%, 31/114), followed by A. dhakensis (17%, 19/114), A. finlandiensis (9%, 10/114), A. caviae (8%, 9/114), A. hydrophila (4%, 4/114), A. jandaei (4%, 4/114), A. rivuli (3%, 3/114), A. enteropelogens (2%, 2/114), A. tecta (2%, 2/114), A. allosaccharophila (1%, 1/114), A. eucrenophila (1%, 1/114), A. media (1%, 1/114) and A. diversa (1%, 1/114). Twenty-six isolates (23%) were unidentifiable at species level. The present study demonstrates that Aeromonas species are highly diverse in freshwater fish tanks, and suggests the potential risks posed by the isolated bacteria to the health of ornamental fish species. [ABSTRACT FROM AUTHOR]
- Published
- 2020
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14. Diverse Profiles of Biofilm and Adhesion Genes in Staphylococcus Aureus Food Strains Isolated from Sushi and Sashimi.
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Puah, Suat Moi, Tan, Jin Ai Mary Anne, Chew, Ching Hoong, and Chua, Kek Heng
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STAPHYLOCOCCUS aureus , *ANTI-infective agents , *ANTIBIOTICS , *STAPHYLOCOCCUS , *BIO-imaging sensors - Abstract
Abstract: Staphylococcus aureus is able to form multilayer biofilms embedded within a glycocalyx or slime layer. Biofilm formation poses food contamination risks and can subsequently increase the risk of food poisoning. Identification of food‐related S. aureus strains will provide additional data on staphylococcal food poisoning involved in biofilm formation. A total of 52 S. aureus strains isolated from sushi and sashimi was investigated to study their ability for biofilm formation using crystal violet staining. The presence of accessory gene regulator (agr) groups and 15 adhesion genes was screened and their associations in biofilm formation were studied. All 52 S. aureus strains showed biofilm production on the tested hydrophobic surface with 44% (23/52) strains classified as strong, 33% (17/52) as moderate, and 23% (12/52) as weak biofilm producers. The frequency of agr‐positive strains was 71% (agr group 1 = 21 strains; agr group 2 = 2 strains; agr group 3 = 12 strains; agr group 4 = 2 strains) whereas agr‐negative strains were 29% (15/52). Twelve adhesion genes were detected and 98% of the S. aureus strains carried at least one adhesion gene. The ebps was significantly (p < .05) associated with strong biofilm producing strains. In addition, eno, clfA, icaAD, sasG, fnbB, cna, and sasC were significantly higher in the agr‐positive group compared to the agr‐negative group. The results of this study suggest that the presence of ebps, eno, clfA, icaAD, sasG, fnbB, cna, and sasC may play an important role in enhancing the stage of biofilm‐related infections and warrants further investigation. Practical Application: This work contributes to the knowledge on the biofilm formation and the distribution of agr groups in S. aureus strains as well as microbial surface components in recognizing adherence matrix molecules of organisms isolated from ready‐to‐eat sushi and sashimi. The findings provide valuable information to further study the roles of specific genes in causing biofilm‐related infections. [ABSTRACT FROM AUTHOR]
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- 2018
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15. Prevalence of Staphylococcus aureus and Salmonella enterica in ready-to-eat sushi and sashimi.
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Suat Moi Puah, Kek Heng Chua, and Tan, Jin Ai Mary Anne
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- 2017
16. Assessment of Risk and Sero-Prevalence of Helicobacter pylori Colonization among Remote Orang Asli Tribes in Peninsula Malaysia.
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Thevakumar, Kavitha, Chandren, Josephine Rebecca, Perez-Perez, Guillermo Ignacio, Chua, Eng Guan, Teh, Lay Kek, Salleh, Mohd Zaki, Tan, Jin Ai Mary Anne, Leow, Alex Hwong Ruey, Goh, Khean Lee, Tay, Alfred Chin Yen, Marshall, Barry J., Vadivelu, Jamuna, Loke, Mun Fai, and Wong, Li Ping
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HELICOBACTER pylori ,ORANG Asal (Malaysian people) ,SOCIOECONOMIC factors ,SEROLOGY ,ANTIGENS - Abstract
The epidemiology of Helicobacter pylori (H. pylori) infection is related to human poverty with marked differences between developing and developed countries. Socioeconomic factors and living standards are the main determinants of the age-dependent acquisition rate of H. pylori, and consequently its prevalence. The aim of this study was to assess the risk and sero-prevalence of H. pylori colonization among Orang Asli in Peninsula Malaysia. This cross-sectional study was conducted on Orang Asli subjects in seven isolated settlements spanning across all three major tribes (Negrito, Proto Malay and Senoi) in Malaysia. Socio-demographic characteristics of the subjects were obtained through interview. Subjects were tested for H. pylori colonization based on CagA and whole cell (WC) antigen serological assays. A total of 275 subjects participated in this study. Among these subjects, 115 (44.7%) were H. pylori sero-positive with highest sero-prevalence among Negrito (65.7%). Among subjects who were H. pylori sero-positive, CagA sero positivity was also significantly higher among Negrito. The highest proportion of respondents reported to be H. pylori sero-positive was from age group 30 years old and below (57.9%), males (56.2%), Negrito (48.6%) and live in bamboo house (92.3%). The highest proportion of respondents reported to be CagA sero-positive was from age group 30 years old and below (41.4%), males (35.6%) and Negrito (48.6%). The results of this study demonstrate that H. pylori colonization can be related to age, gender, tribes and house materials and CagA sero-positive stain closely associated with age, gender and tribes. [ABSTRACT FROM AUTHOR]
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- 2016
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17. Association between plasma soluble RAGE and renal function is unaffected by medication usage and enzymatic antioxidants in chronic kidney disease with type 2 diabetes.
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Wong, Foo Nian, Tan, Jin Ai Mary Anne, Keng, Tee Chau, Ng, Kok Peng, Chua, Kek Heng, and Kuppusamy, Umah Rani
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BLOOD plasma , *GLOMERULAR filtration rate , *SUPEROXIDE dismutase , *TYPE 2 diabetes , *ENZYMATIC analysis , *ANTIOXIDANT analysis , *CHRONIC kidney failure - Abstract
Background This study aimed to investigate the relationship between soluble RAGE and estimated glomerular filtration rate (eGFR) in patients with chronic kidney disease (CKD) after controlling for the potential confounding factors such as medication usage and enzymatic antioxidants. Methods A total of 222 CKD patients whose eGFR is less than 60 ml/min/1.73 m 2 and 111 non-CKD individuals were recruited. The study subjects were classified based on their diabetes status. The plasma glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities as well as plasma soluble RAGE level were measured. Results The plasma GPx and SOD activities were significantly lower and the plasma soluble RAGE level was significantly higher in the CKD patients than in the non-CKD individuals, regardless of the diabetes status. Soluble RAGE was significantly correlated with eGFR in both diabetic CKD (D-CKD) and non-diabetic CKD (ND-CKD) patients. The association between soluble RAGE and eGFR remained largely unaffected by the confounding factors in D-CKD patients. However, the confounding effect of enzymatic antioxidants in the relationship between eGFR and soluble RAGE was observed in ND-CKD patients. Conclusion The increased plasma level of soluble RAGE is a better indicator of renal function decline in diabetic CKD patients instead of non-diabetic CKD patients. [ABSTRACT FROM AUTHOR]
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- 2016
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18. Phenotypic and Genetic Diversity of Aeromonas Species Isolated from Fresh Water Lakes in Malaysia.
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Khor, Wei Ching, Puah, Suat Moi, Tan, Jin Ai Mary Anne, Puthucheary, SD, and Chua, Kek Heng
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PHENOTYPES ,AEROMONAS ,FRESH water ,GLYCEROPHOSPHOLIPIDS - Abstract
Gram-negative bacilli of the genus Aeromonas are primarily inhabitants of the aquatic environment. Humans acquire this organism from a wide range of food and water sources as well as during aquatic recreational activities. In the present study, the diversity and distribution of Aeromonas species from freshwater lakes in Malaysia was investigated using glycerophospholipid-cholesterol acyltransferase (GCAT) and RNA polymerase sigma-factor (rpoD) genes for speciation. A total of 122 possible Aeromonas strains were isolated and confirmed to genus level using the API20E system. The clonality of the isolates was investigated using ERIC-PCR and 20 duplicate isolates were excluded from the study. The specific GCAT-PCR identified all isolates as belonging to the genus Aeromonas, in agreement with the biochemical identification. A phylogenetic tree was constructed using the rpoD gene sequence and all 102 isolates were identified as: A. veronii 43%, A. jandaei 37%, A. hydrophila 6%, A. caviae 4%, A. salmonicida 2%, A. media 2%, A. allosaccharophila 1%, A. dhakensis 1% and Aeromonas spp. 4%. Twelve virulence genes were present in the following proportions—exu 96%, ser 93%, aer 87%, fla 83%, enolase 70%, ela 62%, act 54%, aexT 33%, lip 16%, dam 16%, alt 8% and ast 4%, and at least 2 of these genes were present in all 102 strains. The ascV, aexU and hlyA genes were not detected among the isolates. A. hydrophila was the main species containing virulence genes alt and ast either present alone or in combination. It is possible that different mechanisms may be used by each genospecies to demonstrate virulence. In summary, with the use of GCAT and rpoD genes, unambiguous identification of Aeromonas species is possible and provides valuable data on the phylogenetic diversity of the organism. [ABSTRACT FROM AUTHOR]
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- 2015
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19. Molecular basis of transfusion dependent beta-thalassemia major patients in Sabah.
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Teh, Lai Kuan, George, Elizabeth, Lai, Mei I, Tan, Jin Ai Mary Anne, Wong, Lily, and Ismail, Patimah
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MOLECULAR biology ,BLOOD transfusion ,THALASSEMIA ,PUBLIC health ,DISEASE prevalence ,PATIENTS - Abstract
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. [ABSTRACT FROM AUTHOR]- Published
- 2014
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20. Public perceptions and attitudes toward thalassaemia: Influencing factors in a multi-racial population.
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Wong, Li Ping, George, Elizabeth, and Tan, Jin-Ai Mary Anne
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THALASSEMIA ,SENSORY perception ,ABORTION ,PUBLIC health - Abstract
Background: Thalassaemia is a common public health problem in Malaysia and about 4.5 to 6% of the Malays and Chinese are carriers of this genetic disorder. The major forms of thalassaemia result in death in utero of affected foetuses (a-thalassaemia) or life-long blood transfusions for survival in β-thalassaemia. This study, the first nationwide population based survey of thalassaemia in Malaysia, aimed to determine differences in public awareness, perceptions and attitudes toward thalassaemia in the multi-racial population in Malaysia. Methods: A cross-sectional computer-assisted telephone interview survey of a representative sample of multi-racial Malaysians aged 18 years and above was conducted between July and December 2009. Results: Of a total of 3723 responding households, 2846 (76.4%) have heard of thalassaemia. Mean knowledge score was 11.85 (SD ± 4.03), out of a maximum of 21, with higher scores indicating better knowledge. Statistically significant differences (P < 0.05) in total knowledge score by age groups, education attainment, employment status, and average household income were observed. Although the majority expressed very positive attitudes toward screening for thalassaemia, only 13.6% of married participants interviewed have been screened for thalassaemia. The majority (63.4%) were unsupportive of selective termination of foetuses diagnosed with thalassaemia major. Conclusion: Study shows that carrier and premarital screening programs for thalassaemia may be more effective and culturally acceptable in the reduction of pregnancies with thalassaemia major. The findings provide insights into culturally congruent educational interventions to reach out diverse socio-demographic and ethnic communities to increase knowledge and cultivate positive attitudes toward prevention of thalassaemia. [ABSTRACT FROM AUTHOR]
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- 2011
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21. Dystrophin gene analysis in Duchenne/Becker dystrophy in a Malaysian population using multiplex polymerase chain reaction.
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Tan, Jin-Ai Mary Anne, Chan, James Hsian-Meng, Kim-Lian Tan, Annuar, Azlina Ahmad, Moon-Keen Lee, Khean-Jin Goh, and Kum-Thong Wong
- Subjects
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DYSTROPHIN genes , *GENES , *GENE expression , *GENETIC regulation , *DUCHENNE muscular dystrophy , *BECKER muscular dystrophy , *DIAGNOSTIC use of polymerase chain reaction , *MOLECULAR diagnosis , *GENETICS ,MUSCULAR dystrophy genetics - Abstract
Dystrophinopathy is the commonest form of muscular dystrophy and comprises clinically recognized forms, Duchenne dystrophy and Becker dystrophy. Mutations in the dystrophin gene which consist of large gene deletions (65%), duplications (5%) and point mutations (30%) are responsible for reducing the amount of functional dystrophin protein in skeletal muscle fibres leading to fibre destruction and disease. The aims of this study are to investigate the detection rate, types and distribution of large gene deletions in Malaysian dystrophinopathy patients using the multiplex polymerase chain reaction (MPCR). MPCR of 18 "hot-spot deletion" regions along the dystrophin gene was performed on DNA from 48 muscle biopsy-confirmed cases of dystrophinopathy. A positive detection rate of 58% (28/48) was observed, where 84% (16/19) Indian, 35% (6/17) Chinese and 50% (6/12) Malay ethnic groups showed deletions in their dystrophin genes. The Malaysian Indians appear to have a higher prevalence for large gene deletions compared to the Chinese and Malays. Further analyses of 42 confirmed positive cases (present 28 plus previous 14 cases) by MPCR showed the majority of deletions were in the mid-distal region of the dystrophin gene (81% in exons 45-60). The MPCR is a specific and sensitive method for confirmation of gene deletions responsible for dystrophinopathy. [ABSTRACT FROM AUTHOR]
- Published
- 2010
22. High Prevalence of Alpha- and Beta-Thalassemia in the Kadazandusuns in East Malaysia: Challenges in Providing Effective Health Care for an Indigenous Group.
- Author
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Tan, Jin-Ai Mary Anne, Ping-Chin Lee, Yong-Chui Wee, Kim-Lian Tan, Mahali, Noor Fadzlin, George, Elizabeth, and Kek-Heng Chua
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DISEASE prevalence , *GENETICS of thalassemia , *BETA-Thalassemia , *ALPHA-Thalassemia , *ORANG Asal (Malaysian people) - Abstract
Thalassemia can lead to severe transfusion-dependent anemia, and it is the most common genetic disorder in Malaysia. This paper aims to determine the prevalence of thalassemia in the Kadazandusuns, the largest indigenous group in Sabah, East Malaysia. α- and β-thalassemia were confirmed in 33.6% and 12.8%, of the individuals studied respectively. The high prevalence of α- and β-thalassemia in the Kadazandusuns indicates that thalassemia screening, genetic counseling, and prenatal diagnosis should be included as part of their healthcare system. This preliminary paper serves as a baseline for further investigations into the health and genetic defects of the major indigenous population in Sabah, East Malaysia. [ABSTRACT FROM AUTHOR]
- Published
- 2010
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23. Interaction of Hb South Florida (codon 1; GTG-->ATG) and HbE, with beta-thalassemia (IVS1-1; G-->A): expression of different clinical phenotypes.
- Author
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Tan, Jin-Ai Mary Anne, Tan, Kim-Lian, Omar, Khairul Zaman, Chan, Lee-Lee, Wee, Yong-Chui, and George, Elizabeth
- Subjects
HEMOGLOBINS ,THALASSEMIA in children ,ANEMIA ,GLOBIN genes ,DNA ,ALLELES ,BLOOD transfusion ,COMPARATIVE studies ,DNA probes ,DOCUMENTATION ,GENE expression ,GENES ,GENETICS ,RESEARCH methodology ,MEDICAL cooperation ,GENETIC mutation ,POLYMERASE chain reaction ,RESEARCH ,RNA ,SPLEEN diseases ,PHENOTYPES ,EVALUATION research ,BETA-Thalassemia ,HEPATOMEGALY ,SEQUENCE analysis ,GENOTYPES ,DISEASE complications ,THERAPEUTICS - Abstract
Introduction: Interactions of different hemoglobin variants with thalassemia alleles can result in various clinical phenotypes. HbE-beta-thalassemia generally manifests with severe anemia where individuals exhibit beta-thalassemia major with regular blood transfusions or beta-thalassemia intermedia with periodic blood transfusions. This study presents a unique Malay family with three beta-globin gene defects-HbE, Hb South Florida, and IVS1-1 (G-->A).Materials and Methods: HbE activates a cryptic splice site that produces non-functional mRNAs. Hb South Florida is a rare beta-hemoglobin variant, and its interactions with other beta-thalassemia alleles have not been reported. IVS1-1 is a Mediterranean mutation that affects mRNA processing giving rise to beta(o)-thalassemia.Results and Discussion: Fifteen mutations along the beta-globin gene complex were analyzed using the amplification refractory mutation system. Hb South Florida was identified by direct sequencing using genomic DNA.Conclusion: The affected child with HbE/IVS1-1 produced a beta-thalassemia major phenotype. Compound heterozygosity for Hb South Florida/IVS1-1 produced a beta-thalassemia carrier phenotype in the mother. [ABSTRACT FROM AUTHOR]- Published
- 2009
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24. Thalassemia intermedia in HbH-CS disease with compound heterozygosity for β-thalassemia: Challenges in hemoglobin analysis and clinical diagnosis.
- Author
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Tan, Jin Ai Mary Anne, Kok, Juan Loong, Tan, Kim Lian, Wee, Yong Chui, and George, Elizabeth
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THALASSEMIA ,PHENOTYPES ,GENE amplification ,GENETIC mutation ,BLOOD testing ,MARRIAGE ,CHINESE people - Abstract
Co-inheritance of α-thalassemia with homozygosity or compound heterozygosity for Β-thalassemia may ameliorate Β-thalassemia major. A wide range of clinical phenotypes is produced depending on the number of α-thalassemia alleles (-α/αα--/αα, --/-α). The co-inheritance of Β-thalassemia with α-thalassemia with a single gene deletion (-α/αα) is usually associated with thalassemia major. In contrast, the co-inheritance of Β-thalassemia with two α-genes deleted in cis or trans (--/αα or -α/-α) generally produces Β-thalassemia intermedia. In Southeast Asia, the most common defect responsible for α-thalassemia is the Southeast Asian (SEA) deletion of 20.5 kilobases. The presence of the SEA deletion with Hb Constant Spring (HbCS) produces HbH-CS disease. Co-inheritance of HbH-CS with com-pound heterozygosity for Β-thalassemia is very rare. This study presents a Malay patient with HbH-CS disorder and Β°/Β
+ -thalassemia. The SEA deletion was confirmed in the patient using a duplex-PCR. A Combine-Amplification Refractory Mutation System (C-ARMS) technique to simultaneously detect HbCS and Hb Quong Sze confirmed HbCS in the patient. Compound heterozygosity for CD41/42 and Poly A was confirmed using the ARMS. This is a unique case as the SEA α-gene deletion in cis (--SEA /αα) is generally not present in the Malays, who more commonly posses the two a-gene deletion in trans (-α/-α). In addition, the Β-globin gene mutation at CD41/42 is a common mutation in the Chinese and not in the Malays. The presence of both the SEA deletion and CD41/42 in the mother of the patient suggests the possible introduction of these two defects into the fam-ily by marriage with a Chinese. [ABSTRACT FROM AUTHOR]- Published
- 2009
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25. Heterogeneity in α-thalassemia interactions in Malays, Chinese and Indians in Malaysia.
- Author
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Yong-Chui Wee, Kim-Lian Tan, Teresa Wai-Ping Chow, Sook-Fan Yap, and Tan, Jin-Ai Mary Anne
- Subjects
THALASSEMIA ,FETAL diseases ,HEMOGLOBINS ,GLOBIN genes ,PRENATAL diagnosis - Abstract
Aim: Interactions between different determinants of α-thalassemia raises considerable problems, particularly during pregnancies where antenatal diagnosis is necessary. This study aims to determine the different types of deletional α-thalassemia and Hemoglobin Constant Spring (HbCS), and their frequency in Malays, Chinese and Indians in Malaysia. Methods: DNA from 650 pregnant women from the Antenatal Clinic of the University of Malaya Medical Center in Kuala Lumpur, Malaysia who showed mean cell volume ≤89 fL and/or mean cell hemoglobin ≤28 pg were analyzed for the double α-globin gene South-East Asian deletion (--
SEA ), the -α3.7 and -α4.2 single α-globin gene deletions and HbCS. Results: One hundred and three (15.8%) of the pregnant women were confirmed as α-thalassemia carriers: 25 (3.8%) were α-thalassemia-1 carriers with the --SEA /αα genotype, 64 (9.8%) were heterozygous for the -α3.7 rightward deletion (-α3.7 /αα), four (0.6%) were heterozygous for the -α4.2 leftward deletion (-α4.2 /αα), nine (1.4%) were heterozygous for HbCS (αCS α/αα) and one (0.2%) was compound heterozygous with the -α3.7 /αCS α genotype. The double α-globin gene --SEA deletion was significantly higher in the Chinese (15%) compared to the Malays (2.5%) and not detected in the Indians studied. The -α3.7 deletion was distributed equally in the three races. HbCS and -α4.2 was observed only in the Malays. Conclusion: The data obtained gives a better understanding of the interactions of the different α-thalassemia determinants in the different ethnic groups, thus enabling more rapid and specific confirmation of α-thalassemia in affected pregnancies where antenatal diagnosis is necessary. [ABSTRACT FROM AUTHOR]- Published
- 2005
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26. Higher frequency ofp53gene mutations in diffuse large B-cell lymphoma with MALT component.
- Author
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Tai, Yan-Chin, Tan, Jin-Ai Mary Anne, and Peh, Suat-Cheng
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P53 antioncogene , *GENETIC mutation , *CARCINOGENESIS , *LYMPHOMAS , *GENE expression , *B cells , *IMMUNOHISTOCHEMISTRY , *POLYMERASE chain reaction , *GENETIC polymorphisms - Abstract
p53gene mutation is not a frequent event in the tumorigenesis of lymphomas and the expression of p53 protein is independent ofp53gene mutations. The present study aimed to investigate mutations in thep53gene in a series of extranodal B-cell lymphomas, and its association with p53 protein expression. A total of 52 cases were graded histologically into Grade 1, Grade 2 and Grade 3 tumors and p53 protein expression was detected using immunohistochemistry. Mutations in thep53gene were analyzed using polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) and mobility shifts were confirmed by direct sequencing. The tumors comprised 26 (50%) Grade 1, 9 (17%) Grade 2 and 15 (29%) Grade 3. A high proportion of Grade 2 (25%) tumors expressed p53 protein (P = 0.051) and carriedp53gene mutation (33%) (P = 0.218). However, p53 protein expression was not associated withp53gene mutations (P = 0.057). Transversion mutations (88%) were more frequently detected than transition mutations (12%). The present study revealed thatp53gene mutations and p53 protein expression occurred in higher frequencies in Grade 2 tumors, which may be of pathogenetic importance. The high frequency of transversion mutations may reflect the influence of an etiological agent in the tumorigenesis of mucosa-associated lymphoid tissue (MALT lymphoma). [ABSTRACT FROM AUTHOR]
- Published
- 2004
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27. Mild Beta-Thalassemia intermedia Caused by Compound Heterozygosity for [supG]ϒ([supA]ϒδβ)°/ β-Thalassemia and Molecular Characterization of the Defect in Four Chinese Families.
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Tan Jin Ai, Mary Anne, Sook Fan Yap, Kim Lian Tan, Yean Ching Wong, Yong Chui Wee, and Juan Loong Kok
- Subjects
- *
THALASSEMIA , *GENETIC disorders , *GENETIC polymorphisms , *DNA , *GENETIC mutation - Abstract
Molecular characterization of the compound heterozygous condition--[supG]γ([supA]γδΒ)[supo]/Β-thalassemia--in four families showing mild Β-thalassemia intermedia was carried out using DNA amplification techniques. Using the Amplification Refractory Mutation System (ARMS) to confirm the Β-mutations and DNA amplification to detect the 100-kb Chinese-specific [supG]γ([supA]γδΒ)[supo]-deletion, two families were confirmed to possess [supG]γ([supA]γδΒ)[supo]/&Beta-thalassemia with the IVSII No. 654 Β[sup+]-allele. In the third family, the [supG]γ([supA]γδΒ)[supo]-deletion was confirmed in the father and the mother was a Β-thalassemia carrier with the cd 41-42 Β[supo]-allele. Their affected child with [supG]γ([supA]γδΒ)[supo]/Β-thalassemia was found to be transfusion dependent. The same [supG]γ([supA]γδΒ)[supo]-deletion and Β-thalassemia (cd 41-42) was also confirmed in a fourth family. In addition, the mother was also diagnosed with Hb H disease (genotype-α[sup3.7]/[sup-SEA]). Both the children were found to possess [supG]γ([supA]γδΒ)[supo]/Β-thalassemia but they were not transfusion dependent and this could be due to co-inheritance of α-thalassemia-2 (genotype-α[sup3.7]/αα) in the children together with their compound heterozygous condition. [ABSTRACT FROM AUTHOR]
- Published
- 2003
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- View/download PDF
28. DNA studies are necessary for accurate patient diagnosis in compound heterozygosity for Hb Adana (HBA2:c.179>A) with deletional or nondeletional α-thalassaemia.
- Author
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Tan, Jin Ai Mary Anne, Kho, Siew Leng, Ngim, Chin Fang, Chua, Kek Heng, Goh, Ai Sim, Yeoh, Seoh Leng, and George, Elizabeth
- Published
- 2016
- Full Text
- View/download PDF
29. A novel gap-PCR with high resolution melting analysis for the detection of α-thalassaemia Southeast Asian and Filipino β0-thalassaemia deletion.
- Author
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Leng Kho, Siew, Heng Chua, Kek, George, Elizabeth, and Tan, Jin Ai Mary Anne
- Subjects
THALASSEMIA diagnosis ,ALPHA-Thalassemia ,BETA-Thalassemia ,POLYMERASE chain reaction ,SOUTHEAST Asians ,FILIPINOS ,DISEASES - Abstract
Homozygosity for the α-thalassaemia Southeast Asian (α-SEA) and Filipino β
0 -thalassaemia (β-FIL) deletions can cause serious complications leading to foetal death or life-long blood transfusions. A rapid and accurate molecular detection assay is essential in populations where the deletions are common. In this study, gap-polymerase chain reaction (PCR) with high resolution melting (HRM) analysis was developed to detect both the large deletions. Melting curves at 86.9 ± 0.1 °C were generated by normal individuals without the α-SEA deletion, 84.7 ± 0.1 °C by homozygous α-SEA deletion individuals and two melting curves at 84.7 ± 0.1 °C and 86.9 ± 0.1 °C by α-SEA deletion carriers. Normal individuals without the β-FIL deletion produce amplicons with a melting temperature (Tm) at 74.6 ± 0.1 °C, homozygous β-FIL individuals produce amplicons with Tm at 73.6 ± 0.1 °C and heterozygous β-FIL individuals generate two amplicons with Tm at 73.6 ± 0.1 °C and 74.6 ± 0.1 °C. Evaluation using blinded tests on 220 DNA samples showed 100% sensitivity and specificity. The developed assays are sensitive and specific for rapid molecular and prenatal diagnosis for the α-SEA and β-FIL deletions. [ABSTRACT FROM AUTHOR]- Published
- 2015
- Full Text
- View/download PDF
30. Comparison of Clinical Isolates of <italic>Aeromonas</italic> from Singapore and Malaysia with Regard to Molecular Identification, Virulence, and Antimicrobial Profiles.
- Author
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Khor, Wei Ching, Puah, Suat Moi, Koh, Tse Hsien, Tan, Jin Ai Mary Anne, Puthucheary, Savithiri D., and Chua, Kek Heng
- Subjects
- *
DRUG resistance in bacteria , *MICROBIAL virulence , *ANTI-infective agents , *POLYMERASE chain reaction , *NOSOCOMIAL infections - Abstract
The objective of this study was to examine the species distribution, genetic relatedness, virulence gene profiles, antimicrobial sensitivities, and resistance gene distribution of clinicalObjective: Aeromonas strains from Singapore and Malaysia. A total of 210Methods: Aeromonas clinical isolates were investigated: 116 from Singapore General Hospital and 94 archived clinical isolates from University of Malaya Medical Center, Malaysia. The isolates were genetically identified based on thegcat gene screening and the partial sequences of therpoD housekeeping gene. Genetic relatedness, distribution of 15 virulence genes and 4 beta-lactamase resistance genes, and susceptibility patterns to 11 antimicrobial agents were compared. Of the 210Results: Aeromonas isolates,A. dhakensis– 94 (45%) was the dominant species in Singapore and Malaysia. Species composition was similar and enterobacterial repetitive intergenic consensus-PCR did not show genetic relatedness between strains from the two countries. Of the 15 virulence genes,A. dhakensis andA. hydrophila harbored the most compared with other species. Different combinations of 9 virulence genes (exu ,fla ,lip ,eno ,alt ,dam ,hlyA ,aexU, andascV ) were present inA. dhakensis ,A. hydrophila , andA. veronii from both the countries. Distribution of virulence genes was species and anatomic site related. Majority (>80%) of the strains were susceptible to all antimicrobial agents tested, except amoxicillin and cephalothin.A. dhakensis strains from Malaysia significantly harbored thecphA gene compared withA. dhakensis from Singapore. Multidrug resistance was mostly detected in strains from peritoneal fluids of dialysis patients. This study revealedConclusion: A. dhakensis as the dominant species isolated in both geographic regions, and that it carried a high number of virulence genes. It also highlights the geographic-related differences of virulence gene distribution and antimicrobial resistance profiles of clinicalAeromonas strains from Singapore and Malaysia. [ABSTRACT FROM AUTHOR]- Published
- 2018
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31. Development of a species-specific PCR-RFLP targeting rpoD gene fragment for discrimination of Aeromonas species.
- Author
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Puah SM, Khor WC, Kee BP, Tan JAMA, Puthucheary SD, and Chua KH
- Subjects
- Aeromonas classification, Aeromonas isolation & purification, Aeromonas metabolism, Bacterial Proteins metabolism, Bacterial Typing Techniques, DNA Primers genetics, Polymorphism, Restriction Fragment Length, Sigma Factor metabolism, Species Specificity, Aeromonas genetics, Bacterial Proteins genetics, Polymerase Chain Reaction methods, Sigma Factor genetics
- Abstract
Purpose: The taxonomy of Aeromonas keeps expanding and their identification remains problematic due to their phenotypic and genotypic heterogeneity. In this study, we aimed to develop a rapid and reliable polymerase chain reaction-restriction fragment length polymorphism assay targeting the rpoD gene to enable the differentiation of aeromonads into 27 distinct species using microfluidic capillary electrophoresis., Methodology: A pair of degenerate primers (Aero F: 5'-YGARATCGAYATCGCCAARCGB-3' and Aero R: 5'-GRCCDATGCTCATRCGRCGGTT-3') was designed that amplified the rpoD gene of 27 Aeromonas species. Subsequently, in silico analysis enabled the differentiation of 25 species using the single restriction endonuclease AluI, while 2 species, A. sanarelli and A. taiwanensis, required an additional restriction endonuclease, HpyCH4IV. Twelve type strains (A. hydrophila ATCC7966
T , A. caviae ATCC15468T , A. veronii ATCC9071T , A. media DSM4881T , A. allosaccharophila DSM11576T , A. dhakensis DSM17689T , A. enteropelogens DSM7312T , A. jandaei DSM7311T , A. rivuli DSM22539T , A. salmonicida ATCC33658T , A. taiwanensis DSM24096T and A. sanarelli DSM24094T ) were randomly selected from the 27 Aeromonas species for experimental validation.Results/key findings. The twelve type strains demonstrated distinctive RFLP patterns and supported the in silico digestion. Subsequently, 60 clinical and environmental strains from our collection, comprising nine Aeromonas species, were used for screening examinations, and the results were in agreement., Conclusion: This method provides an alternative method for laboratory identification, surveillance and epidemiological investigations of clinical and environmental specimens.- Published
- 2018
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- View/download PDF
32. Glycaemic control in type 2 diabetic patients with chronic kidney disease: the impacts on enzymatic antioxidants and soluble RAGE.
- Author
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Wong FN, Chua KH, Tan JAMA, Wong CM, and Kuppusamy UR
- Abstract
Background: Chronic kidney disease (CKD) is characterised by long-term kidney damage and renal function decline. Diabetic CKD is the principal subtype of kidney disease in Malaysia and is associated with oxidative stress which plays an important role in development and progression of the disease. Glycaemic control slows down the progression of diabetic complications, including diabetic CKD. However, the implication of glycaemic control on enzymatic antioxidants and soluble RAGE (sRAGE) in CKD patients remains elusive. The aim of this study was to investigate the effect of glycaemic control on the levels or activities of glutathione peroxidase (GPx), superoxide dismutase (SOD) and sRAGE in CKD patients., Methods: A total of 150 CKD patients and 64 non-CKD patients were enrolled. The type 2 diabetic patients in the recruited study participants were categorised based on their glycaemic control; poor glycaemic control (GC) with haemoglobin A1c (HbA1c) > 7% and good GC with HbA1c ≤ 7%. The levels or activities of GPx, SOD and sRAGE in plasma were measured. These biochemical parameters were analysed using Mann-Whitney U test and two-way analysis of variance (ANOVA)., Results: The activities of GPx and SOD as well as plasma level of sRAGE were not significantly different among the CKD patients with varying glycaemic control status. Irrespective of diabetes status and glycaemic control status, CKD patients also exhibited lower plasma SOD activities compared with non-CKD patients. Among the non-CKD patients, SOD activities were significantly higher in diabetic patients with good GC than diabetic patients with poor GC. Two-way ANOVA revealed that both CKD status and glycaemic control had an interaction effect on SOD activities in diabetic subjects with and without CKD. Follow-up analysis showed that SOD activities were significantly higher in non-CKD patients with good GC. There were no overall significant differences in GPx activities among the study participants. Furthermore, plasma sRAGE levels were higher in diabetic patients with CKD than those without CKD, regardless of glycaemic control status. There were no interaction effects between CKD status and glycaemic control status on GPx and sRAGE. Instead, CKD status showed significant main effects on these parameters, indicating significant differences between diabetic subjects with CKD and diabetic subjects without CKD., Conclusion: Glycaemic control did not quantitatively alter GPx, SOD and sRAGE in diabetic CKD patients. Despite the advantages of good glycaemic control, a well-controlled diabetes in CKD did not modulate the activities of enzymatic antioxidants and sRAGE levels, therefore may not be the primary mechanism to handle oxidative stress., Competing Interests: The authors declare there are no competing interests.
- Published
- 2018
- Full Text
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33. Association of the receptor for advanced glycation end-products (RAGE) gene polymorphisms in Malaysian patients with chronic kidney disease.
- Author
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Wong FN, Chua KH, Kuppusamy UR, Wong CM, Lim SK, and Tan JA
- Abstract
Background: Chronic kidney disease (CKD) is a condition associated with progressive loss of kidney function and kidney damage. The two common causes of CKD are diabetes mellitus and hypertension. Other causes of CKD also include polycystic kidney disease, obstructive uropathy and primary glomerulonephritis. The receptor for advanced glycation end-products (RAGE) is a multi-ligand cell surface receptor of the immunoglobulin superfamily and it has been associated with kidney disease in both non-diabetic and diabetic patients. Presently, data on the association between RAGE polymorphisms and CKD in the Malaysian population is limited, while numerous studies have reported associations of RAGE polymorphisms with diabetic complications in other populations. The present study aims to explore the possibility of using RAGE polymorphisms as candidate markers of CKD in Malaysian population by using association analysis., Methods: A total of 102 non-diabetic CKD patients, 204 diabetic CKD patients and 345 healthy controls were enrolled in the study. DNA isolated from blood samples were subjected to genotyping of RAGE G82S, -374T/A, -429T/C, 1704G/T and 2184A/G polymorphisms using real-time polymerase chain reaction (PCR). The 63-bp deletion, a polymorphism in the RAGE gene promoter, was genotyped using conventional PCR method and visualized using agarose gel electrophoresis. The collective frequencies of genotypes with at least one copy of the minor alleles of the four polymorphisms were compared between the non-diabetic CKD patients, diabetic CKD patients and healthy controls., Results: After adjustment of age, gender and ethnic groups in binary logistic regression analysis, the G82S CT + TT genotypes were associated with non-diabetic CKD patients when compared with diabetic CKD patients (p = 0.015, OR = 1.896, 95% CI = 1.132-3.176). After further adjustment of CKD comorbidities, the G82S CT + TT genotypes were still associated with non-diabetic CKD patients when compared with diabetic CKD patients (p = 0.011, OR = 2.024, 95% CI = 1.178-3.476). However, it cannot be suggested that G82S polymorphism was associated with CKD in non-diabetic patients in this study. This is because there were no significant differences in the frequencies of G82S CT + TT genotypes between non-diabetic CKD patients and healthy controls. In addition, the RAGE -374T/A, -429T/C, 1704G/T, 2184A/G and 63-bp deletion polymorphisms were also not associated with non-diabetic CKD patients and diabetic CKD patients in this study., Conclusion: The G82S, -374T/A, -429T/C, 1704G/T, 2184A/G and 63-bp deletion polymorphisms examined in this study were not associated with chronic kidney disease in the Malaysian patients.
- Published
- 2016
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34. Virulence Factors and Antibiotic Susceptibility of Staphylococcus aureus Isolates in Ready-to-Eat Foods: Detection of S. aureus Contamination and a High Prevalence of Virulence Genes.
- Author
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Puah SM, Chua KH, and Tan JA
- Subjects
- Animals, Drug Resistance, Multiple, Bacterial drug effects, Enterotoxins, Humans, Prevalence, Staphylococcal Infections epidemiology, Anti-Bacterial Agents pharmacology, Fishes microbiology, Food Supply, Microbial Sensitivity Tests, Staphylococcus aureus drug effects, Staphylococcus aureus isolation & purification, Virulence genetics, Virulence Factors genetics
- Abstract
Staphylococcus aureus is one of the leading causes of food poisoning. Its pathogenicity results from the possession of virulence genes that produce different toxins which result in self-limiting to severe illness often requiring hospitalization. In this study of 200 sushi and sashimi samples, S. aureus contamination was confirmed in 26% of the food samples. The S. aureus isolates were further characterized for virulence genes and antibiotic susceptibility. A high incidence of virulence genes was identified in 96.2% of the isolates and 20 different virulence gene profiles were confirmed. DNA amplification showed that 30.8% (16/52) of the S. aureus carried at least one SE gene which causes staphylococcal food poisoning. The most common enterotoxin gene was seg (11.5%) and the egc cluster was detected in 5.8% of the isolates. A combination of hla and hld was the most prevalent coexistence virulence genes and accounted for 59.6% of all isolates. Antibiotic resistance studies showed tetracycline resistance to be the most common at 28.8% while multi-drug resistance was found to be low at 3.8%. In conclusion, the high rate of S. aureus in the sampled sushi and sashimi indicates the need for food safety guidelines.
- Published
- 2016
- Full Text
- View/download PDF
35. A novel gap-PCR with high resolution melting analysis for the detection of α-thalassaemia Southeast Asian and Filipino β°-thalassaemia deletion.
- Author
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Kho SL, Chua KH, George E, and Tan JA
- Subjects
- Asian People, DNA analysis, DNA metabolism, Humans, Philippines, Real-Time Polymerase Chain Reaction, Transition Temperature, alpha-Thalassemia genetics, beta-Thalassemia genetics, alpha-Thalassemia diagnosis, beta-Globins genetics, beta-Thalassemia diagnosis
- Abstract
Homozygosity for the α-thalassaemia Southeast Asian (α-SEA) and Filipino β°-thalassaemia (β-FIL) deletions can cause serious complications leading to foetal death or life-long blood transfusions. A rapid and accurate molecular detection assay is essential in populations where the deletions are common. In this study, gap-polymerase chain reaction (PCR) with high resolution melting (HRM) analysis was developed to detect both the large deletions. Melting curves at 86.9 ± 0.1 °C were generated by normal individuals without the α-SEA deletion, 84.7 ± 0.1 °C by homozygous α-SEA deletion individuals and two melting curves at 84.7 ± 0.1 °C and 86.9 ± 0.1 °C by α-SEA deletion carriers. Normal individuals without the β-FIL deletion produce amplicons with a melting temperature (Tm) at 74.6 ± 0.1 °C, homozygous β-FIL individuals produce amplicons with Tm at 73.6 ± 0.1 °C and heterozygous β-FIL individuals generate two amplicons with Tm at 73.6 ± 0.1 °C and 74.6 ± 0.1 °C. Evaluation using blinded tests on 220 DNA samples showed 100% sensitivity and specificity. The developed assays are sensitive and specific for rapid molecular and prenatal diagnosis for the α-SEA and β-FIL deletions.
- Published
- 2015
- Full Text
- View/download PDF
36. Non-Invasive DNA Sampling for Molecular Analysis of Beta-Thalassemia: Amiable Alternative Sampling Methods with Accurate Results for Pediatric Patients.
- Author
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Abd Rahim MR, Kho SL, Kuppusamy UR, and Tan JA
- Subjects
- Adolescent, Child, Child, Preschool, DNA genetics, Epithelial Cells chemistry, Female, Humans, Male, Mouthwashes, Nucleic Acid Amplification Techniques, Specimen Handling instrumentation, Therapeutic Irrigation, beta-Thalassemia metabolism, DNA isolation & purification, DNA Mutational Analysis methods, Molecular Diagnostic Techniques methods, Mouth Mucosa chemistry, Saliva chemistry, Specimen Handling methods, beta-Globins genetics, beta-Thalassemia genetics
- Abstract
Background: Beta-thalassemia is the most common genetic disorder in Malaysia. Confirmation of the β-globin gene mutations involved in thalassemia is usually carried out by molecular analysis of DNA extracted from leukocytes in whole blood. Molecular analysis is generally carried out when affected children are around 1 - 2 years as clinical symptoms are expressed during this period. Blood taking at this age can be distressing for the child. High yield and pure DNA extracted from non-invasive sampling methods can serve as alternative samples in molecular studies for genetic diseases especially in pediatric cases., Methods: In this study, mouthwash, saliva, and buccal cytobrush samples were collected from β-thalassemia major patients who had previously been characterized using DNA extracted from peripheral blood. DNA was extracted from mouthwash, saliva, and buccal cytobrush samples using the conventional inexpensive phenol-chloroform method and was measured by spectrophotometry for yield and purity. Molecular characterization of β-globin gene mutations was carried out using the amplification refractory mutation system (ARMS)., Results: DNA extracted from mouthwash, saliva, and buccal cytobrush samples produced high concentration and pure DNA. The purified DNA was successfully amplified using ARMS. Results of the β-globin gene mutations using DNA from the three non-invasive samples were in 100% concordance with results from DNA extracted from peripheral blood., Conclusions: The conventional in-house developed methods for non-invasive sample collection and DNA extraction from these samples are effective and negate the use of more expensive commercial kits. In conclusion, DNA extracted from mouthwash, saliva, and buccal cytobrush samples provided sufficiently high amounts of pure DNA suitable for molecular analysis of β-thalassemia.
- Published
- 2015
- Full Text
- View/download PDF
37. High throughput molecular confirmation of β-thalassemia mutations using novel TaqMan probes.
- Author
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Kho SL, Chua KH, George E, and Tan JA
- Subjects
- Alleles, Base Sequence, DNA genetics, DNA Mutational Analysis, Genotyping Techniques, Humans, Reproducibility of Results, Templates, Genetic, DNA Probes metabolism, High-Throughput Screening Assays methods, Mutation genetics, Taq Polymerase metabolism, beta-Thalassemia genetics
- Abstract
β-Thalassemia is a public health problem where 4.5% of Malaysians are β-thalassemia carriers. The genetic disorder is caused by defects in the β-globin gene complex which lead to reduced or complete absence of β-globin chain synthesis. Five TaqMan genotyping assays were designed and developed to detect the common β-thalassemia mutations in Malaysian Malays. The assays were evaluated with 219 "blinded" DNA samples and the results showed 100% sensitivity and specificity. The in-house designed TaqMan genotyping assays were found to be cost- and time-effective for characterization of β-thalassemia mutations in the Malaysian population.
- Published
- 2013
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38. Molecular characterisation of Haemoglobin Constant Spring and Haemoglobin Quong Sze with a Combine-Amplification Refractory Mutation System.
- Author
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Wee YC, Tan KL, Chua KH, George E, and Tan JA
- Abstract
Background: The interaction of the non-deletional α(+)-thalassaemia mutations Haemoglobin Constant Spring and Haemoglobin Quong Sze with the Southeast Asian double α-globin gene deletion results in non-deletional Haemoglobin H disease. Accurate detection of non-deletional Haemoglobin H disease, which is associated with severe phenotypes, is necessary as these mutations have been confirmed in the Malaysian population., Methods: DNA from two families with Haemoglobin H disease was extracted from EDTA-anticoagulated whole blood and subjected to molecular analysis for α-thalassaemia. A duplex polymerase chain reaction was used to detect the Southeast Asian α-globin gene deletion. Polymerase chain reaction-restriction fragment length polymorphism analysis was then carried out to determine the presence of Haemoglobin Constant Spring and Haemoglobin Quong Sze. A combine-amplification refractory mutation system protocol was optimised and implemented for the rapid and specific molecular characterisation of Haemoglobin Constant Spring and Haemoglobin Quong Sze in a single polymerase chain reaction., Results and Conclusions: The combine-amplification refractory mutation system for Haemoglobin Constant Spring and Haemoglobin Quong Sze, together with the duplex polymerase chain reaction, provides accurate pre- and postnatal diagnosis of non-deletional Haemoglobin H disease and allows detailed genotype analyses using minimal quantities of DNA.
- Published
- 2009
39. Thalassemia intermedia in HbH-CS disease with compound heterozygosity for beta-thalassemia: challenges in hemoglobin analysis and clinical diagnosis.
- Author
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Tan JA, Kok JL, Tan KL, Wee YC, and George E
- Subjects
- Asian People, Base Sequence genetics, DNA Mutational Analysis methods, Female, Humans, Infant, Malaysia, Male, Pedigree, Sequence Deletion, alpha-Thalassemia complications, beta-Thalassemia complications, Hemoglobins, Abnormal genetics, alpha-Thalassemia diagnosis, alpha-Thalassemia genetics, beta-Thalassemia diagnosis, beta-Thalassemia genetics
- Abstract
Co-inheritance of alpha-thalassemia with homozygosity or compound heterozygosity for beta-thalassemia may ameliorate beta-thalassemia major. A wide range of clinical phenotypes is produced depending on the number of alpha-thalassemia alleles (-alpha/alphaalpha --/alphaalpha, --/-alpha). The co-inheritance of beta-thalassemia with alpha-thalassemia with a single gene deletion (-alpha/alphaalpha) is usually associated with thalassemia major. In contrast, the co-inheritance of beta-thalassemia with two alpha-genes deleted in cis or trans (--/alphaalpha or -alpha/-alpha) generally produces beta-thalassemia intermedia. In Southeast Asia, the most common defect responsible for alpha-thalassemia is the Southeast Asian (SEA) deletion of 20.5 kilobases. The presence of the SEA deletion with Hb Constant Spring (HbCS) produces HbH-CS disease. Co-inheritance of HbH-CS with compound heterozygosity for beta-thalassemia is very rare. This study presents a Malay patient with HbH-CS disorder and beta degrees/beta+-thalassemia. The SEA deletion was confirmed in the patient using a duplex-PCR. A Combine-Amplification Refractory Mutation System (C-ARMS) technique to simultaneously detect HbCS and Hb Quong Sze confirmed HbCS in the patient. Compound heterozygosity for CD41/42 and Poly A was confirmed using the ARMS. This is a unique case as the SEA alpha-gene deletion in cis (--SEA/alphaalpha) is generally not present in the Malays, who more commonly possess the two alpha-gene deletion in trans (-alpha/-alpha). In addition, the beta-globin gene mutation at CD41/42 is a common mutation in the Chinese and not in the Malays. The presence of both the SEA deletion and CD41/42 in the mother of the patient suggests the possible introduction of these two defects into the family by marriage with a Chinese.
- Published
- 2009
- Full Text
- View/download PDF
40. Molecular characterisation and frequency of Ggamma Xmn I polymorphism in Chinese and Malay beta-thalassaemia patients in Malaysia.
- Author
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Wong YC, George E, Tan KL, Yap SF, Chan LL, and Tan JA
- Subjects
- Asian People, Humans, Malaysia ethnology, Deoxyribonucleases, Type II Site-Specific genetics, Polymerase Chain Reaction methods, Polymorphism, Genetic, beta-Thalassemia genetics
- Abstract
The molecular basis of variable phenotypes in P-thalassaemia patients with identical genotypes has been associated with co-inheritance of alpha-thalassaemia and persistence of HbF production in adult life. The Xmn I restriction site at -158 position of the Ggamma-gene is associated with increased expression of the Ggamma-globin gene and higher production of HbF This study aims to determine the frequency of the digammaferent genotypes of the Ggamma Xmn I polymorphism in P-thalassaemia patients in two ethnic groups in Malaysia. Molecular characterisation and frequency of the Ggamma Xmn I polymorphism were studied in fifty-eight Chinese and forty-nine beta-thalassaemia Malay patients by Xmn I digestion after DNA amplification of a 650 bp sequence. The in-house developed technique did not require further purification or concentration of amplified DNA before restriction enzyme digestion. The cheaper Seakem LE agarose was used instead of Nusieve agarose and distinct well separated bands were observed. Genotyping showed that the most frequent genotype observed in the Malaysian Chinese was homozygosity for the absence of the Xmn I site (-/-) (89.7%). In the Malays, heterozygosity of the Xmn I site (+/-) was most common (63.3%). Homozygosity for the Xmn I site (+/+) was absent in the Chinese, but was confirmed in 8.2% of the Malays. The ratio of the (+) allele (presence of the Xmn I site) to the (-) allele (absence of the Xmn I site)) was higher in the Malays (0.66) compared to the Chinese (0.05). The (+/-) and (+/+) genotypes are more commonly observed in the Malays than the Chinese in Malaysia.
- Published
- 2006
41. Heterogeneity in alpha-thalassemia interactions in Malays, Chinese and Indians in Malaysia.
- Author
-
Wee YC, Tan KL, Chow TW, Yap SF, and Tan JA
- Subjects
- Adolescent, Adult, Base Sequence, DNA genetics, Female, Genetic Carrier Screening, Globins genetics, Humans, Malaysia, Pregnancy, Pregnancy Complications, Hematologic diagnosis, alpha-Thalassemia diagnosis, alpha-Thalassemia prevention & control, Asian People genetics, Pregnancy Complications, Hematologic genetics, alpha-Thalassemia complications, alpha-Thalassemia genetics
- Abstract
Aim: Interactions between different determinants of alpha-thalassemia raises considerable problems, particularly during pregnancies where antenatal diagnosis is necessary. This study aims to determine the different types of deletional alpha-thalassemia and Hemoglobin Constant Spring (HbCS), and their frequency in Malays, Chinese and Indians in Malaysia., Methods: DNA from 650 pregnant women from the Antenatal Clinic of the University of Malaya Medical Center in Kuala Lumpur, Malaysia who showed mean cell volume < or =89 fL and/or mean cell hemoglobin < or =28 pg were analyzed for the double alpha-globin gene South-East Asian deletion (--SEA), the -alpha3.7 and -alpha4.2 single alpha-globin gene deletions and HbCS., Results: One hundred and three (15.8%) of the pregnant women were confirmed as alpha-thalassemia carriers: 25 (3.8%) were alpha-thalassemia-1 carriers with the --SEA/alphaalpha genotype, 64 (9.8%) were heterozygous for the -alpha3.7 rightward deletion (-alpha3.7/alphaalpha), four (0.6%) were heterozygous for the -alpha4.2 leftward deletion (-alpha4.2/alphaalpha), nine (1.4%) were heterozygous for HbCS (alphaCSalpha/alphaalpha) and one (0.2%) was compound heterozygous with the -alpha3.7/alphaCSalpha genotype. The double alpha-globin gene --SEA deletion was significantly higher in the Chinese (15%) compared to the Malays (2.5%) and not detected in the Indians studied. The -alpha3.7 deletion was distributed equally in the three races. HbCS and -alpha4.2 was observed only in the Malays., Conclusion: The data obtained gives a better understanding of the interactions of the different alpha-thalassemia determinants in the different ethnic groups, thus enabling more rapid and specific confirmation of alpha-thalassemia in affected pregnancies where antenatal diagnosis is necessary.
- Published
- 2005
- Full Text
- View/download PDF
42. The use of the amplification refractory mutation system (arms) in the detection of rare beta-thalassemia mutations in the Malays and Chinese in Malaysia.
- Author
-
Chan Yoke Fan, Tan Kim Lian, Wong Yean Ching, Wee Yong Chui, Yap Sook Fan, and Tan Jin Ai Mary Anne
- Subjects
- Base Sequence, China ethnology, DNA Primers, Female, Humans, Malaysia, Male, Pedigree, Transcription, Genetic, beta-Thalassemia diagnosis, Mutation, Polymerase Chain Reaction methods, beta-Thalassemia genetics
- Abstract
Molecular characterization and prenatal diagnosis for beta-thalassemia can be carried out using the Amplification Refractory Mutation System (ARMS). The ARMS is a rapid and direct molecular technique in which beta-thalassemia mutations are visualized immediately after DNA amplification by gel electrophoresis. In the University of Malaya Medical Center, molecular characterization and prenatal diagnosis for beta-thalassemia is carried out using ARMS for about 96% of the Chinese and 84.6% of the Malay patients. The remaining 4% and 15.4% of the uncharacterized mutations in the Chinese and Malay patients respectively are detected using DNA sequencing. DNA sequencing is an accurate technique but it is more time-consuming and expensive compared with the ARMS. The ARMS for the rare Chinese beta-mutations at position -29 (A-->G) and the ATG-->AGG base substitution at the initiator codon for translation in the beta-gene was developed. In the Malays, ARMS was optimized for the beta-mutations at codon 8/9 (+G), Cap (+1) (A-->C) and the AATAAA-->AATAGA base substitution in the polyadenylation region of the beta-gene. The ARMS protocols were developed by optimization of the parameters for DNA amplification to ensure sensitivity, specificity and reproducibility. ARMS primers (sequences and concentration), magnesium chloride concentration, Taq DNA polymerase and PCR cycling parameters were optimized for the specific amplification of each rare beta-thalassemia mutation. The newly-developed ARMS for the 5 rare beta-thalassemia mutations in the Chinese and Malays in Malaysia will allow for more rapid and cost-effective molecular characterization and prenatal diagnosis for beta-thalassemia in Malaysia.
- Published
- 2001
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